No.

204

ADVISORY COMMITTEE ON PESTICIDES

FOOD AND ENVIRONMENT PROTECTION ACT 1985, PART III Control of Pesticides Regulations 1986

Evaluation of Fully Approved or Provisionally Approved Products Evaluation on: CHLOROTHALONIL: USE AS A BOOSTER BIOCIDE IN ANTIFOULING PRODUCTS

AUGUST 2002

Prepared by : The Health and Safety Executive Biocides & Pesticides Assessment Unit Magdalen House Stanley Precinct Bootle Merseyside L20 3QZ Available from: Department for Environment, Food and Rural Affairs Pesticides Safety Directorate Mallard House Kings Pool 3 Peasholme Green York YO1 7PX

IMPORTANT NOTICE TO RECIPIENTS OF EVALUATION DOCUMENTS PUBLISHED BY THE ADVISORY COMMITTEE ON PESTICIDES

In making this information available, the Advisory Committee on Pesticides (ACP) Secretariat is obliged to consider its commercial value. You are therefore required, under regulation 8(4) of The Control of Pesticides Regulations 1986, as amended by the Control of Pesticides (Amendment) Regulations 1997, not to make commercial use of the contents of this publication nor publish any part unless authorised. Should you wish to publish all or part of this document, requests for written authorisation should be addressed to: ACP Secretariat, Room 202 Mallard House, 3 Peasholme Green, YORK YO1 7PX The Control of Pesticides Regulations 1986 (SI 1986/1510) and the Control of Pestcides (Amendment) Regulations 1997 (SI 1997/188) are available from the Stationery Office. Public access to raw data underlying these publications can be arranged by contacting: The Biocides & Pesticides Assessment Unit HSE Magdalen House Stanley Precinct Bootle Merseyside L20 3QZ.

NB: This document reflects the outcome of the 278th Advisory Committee on Pesticides meetings held in September 2000.

CONTENTS
LIST OF ABBREVIATIONS OVERALL SUMMARY 1 2 3 4 5 6 7 8 9 ii iv 1 7 11 22 44 74 92 99 109 113 118 123

INTRODUCTION AND REGISTRATION HISTORY PHYSICAL CHEMISTRY OF CHLOROTHALONIL MAMMALIAN TOXICOKINETICS AND TOXICOLOGY OPERATOR AND CONSUMER EXPOSURE AND RISK ASSESSMENTS ENVIRONMENTAL FATE AND BEHAVIOUR ECOTOXICOLOGY ENVIRONMENTAL RISK ASSESSMENT EFFICACY OVERALL RECOMMENDATIONS AND DATA REQUIREMENTS

References Appendix 1 Appendix 2

LIST OF ABBREVIATIONS
AChE ACP AFP(s) ai AOEL AR ASTM BCF BSI 14 C CAS CEFAS CEPE CL CO2 COPR COSHH d DCNB DEFRA DETR DMF EC50 ECD EINECS FEV FID FIFRA g GC g kg-1 GLP GSH GST h HSE HPLC i.d. IR IUPAC kPa LC50 LD50 LOEC LSC MAFF mg ai l-1 acetyl cholinesterase Advisory Committee on Pesticides antifouling product(s) active ingredient admissible operator exposure level applied radioactivity American Society for Testing and Materials bioconcentration factor British Standards Institute radiolabelled carbon Chemical Abstract Services Centre for Environment, Fisheries & Aquaculture Science European Confederation of Paint, Printers Inks and Artists Colours Manufacturers Association Confidence Limits carbon dioxide The Control of Pesticide Regulations (1986) Control of Substances Hazardous to Health day (s) dinitrochlorobenzene Department of Environment, Food and Rural Affairs Department of Environment, Transport and the Regions Dimethylformamide effective concentration (50%) electron capture detector European Inventory of Existing Commercial Chemical Substances forced expiratory volume Flame Ionisation Detector Federal Insecticide, Fungicide and Rodenticide Act (US) gram (s) gas chromatography grams per kilogram Good Laboratory Practice glutathione glutathione-S-tranferase hour (s) Health and Safety Executive high performance liquid chromatograph internal diameter infra-red spectroscopy International Union of Pure and Applied Chemistry kilopascals lethal concentration (50%) lethal dose (50%) lowest observed effect concentration liquid scintillation counting Ministry of Agriculture Fisheries and Food milligrams of active ingredient per litre ii

mg kg-1 mg kg-1 bw mg l-1 MIC min ml ml l-1 mm mPa MS nm NMR NOEC NOEL OECD Pa PEC PECsediment PECwater PNEC Pow ppm QWASI REMA SCP SPC STP TBT TBTO TLC T US EPA UV v/v w w/w y g l-1 l m

milligrams per kilogram milligrams per kilogram of body weight milligrams per litre minimum inhibitory concentration minute (s) millilitre (s) millilitres per litre millimetre (s) millipascals mass spectra nanometre (s) nuclear magnetic resonance No Observed Effect Concentration no observed effect level Organisation for Economic Co-operation and Development Pascals predicted environmental concentrations predicted environmental concentrations in sediment predicted environmental concentrations in water predicted no-effect concentration octanol/water partition coefficient parts per million quantitative water air sediment interface Regulatory Environmental Modelling of Antifoulants Scientific Subcommittee on Pesticides self polishing co-polymer sewage treatment processes tributyltin tributyltin oxide thin layer chromatography half-life United States Environmental Protection Agency ultra violet volume for volume week (s) weight for weight year (s) micrograms per litre microlitre (s) micrometre (s)

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OVERALL SUMMARY
Introduction Chlorothalonil is used as a fungicide in agriculture and as an active ingredient in approved professional and amateur antifouling products for use on vessels of any size plus structures below the waterline. In September 2000, all 4 of the currently approved antifouling products containing chlorothalonil were intended for amateur and professional use. Although products may contain up to 5.0 % w/w of the compound, the current maximum level in existing products is only 3.0 % w/w. Acceptable application methods for applying approved products containing chlorothalonil to vessels/structures are by brush, roller and airless or conventional spray. In 1995, the ACP considered reviews of triorganotin compounds (TOTs) in antifouling products and copper compounds. It concluded that the risk from the use of TOTs was unacceptable but that the use of copper antifouling products (AFPs) was acceptable. However the ACP recommended that a high priority should be placed on reviewing the organic booster biocides since restrictions on the use of TOT-containing products could lead to an increased environmental loading of these compounds. Booster biocides is a generic term given to a group of compounds normally found in addition to copper and occasionally TOT, in AFPs. The purpose of these compounds is to enhance the products efficacy against a broader spectrum of fouling organisms than that achieved with copper alone. Booster biocides are either organic or organo-metallic compounds which have fungicidal, herbicidal or anti-microbial actions. A partial review of chlorothalonil was presented to the ACP in 1996 when further data requirements were set. The ACP considered the full review of chlorothalonil at its 278th meeting in September 2000. This document comprises a summary of data presented in earlier reviews together with an evaluation of new data submitted in response to earlier data requirements. An update of the published literature is also included. The ACP recommended that all amateur uses of antifouling products containing chlorothalonil should be revoked because information from humans indicates that the risk of skin sensitisation is unacceptable. Approvals for the professional use of antifouling products containing chlorothalonil at a maximum concentration of 5 % w/w should be continued for professional use by brush, roller and spray, subject to the data requirements and conditions detailed at the end of this summary and in Chapter 9. Ministers accepted these recommendations. Physical Chemistry Tetrachloroisophthalonitrile (IUPAC) or chlorothalonil (BSI name) has a purity of 985 g kg1 . It is a white powder of molecular mass 265.9; melting point 252.5 - 254.5oC (pure chlorothalonil); boiling point ~350 oC; relative density 1.73 (at 20 oC); and vapour pressure 0.22 mPa (at 25 oC); it is sparingly soluble in water (0.54 mg l-1 at 20 oC) with log Pow 2.93 (at 22 oC). Analytical methods have been provided for determination of the active ingredient and impurities in technical chlorothalonil; and for the determination of chlorothalonil in water.

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Mammalian Toxicokinetics and Toxicology Following oral dosing chlorothalonil was absorbed and rapidly eliminated by the rat with a half life of 6 to 7 hours at dose levels of 50 mg kg-1 or less. Excretion in the bile accounted for a greater proportion of the dose than excretion in the urine. There was evidence to suggest that entero hepatic recirculation occurs. In an in vitro study it was shown that chlorothalonil is only absorbed from the duodenum as glutathione conjugates. In a study with germ-free rats the absorption of chlorothalonil was greatly reduced compared with non germ-free rats suggesting that the gut flora plays an important role in the absorption of chlorothalonil. In a rat dermal absorption study 6 % of a 5 mg kg-1 dose was absorbed in 8 hours. Metabolism studies showed that the main metabolites formed were the mono- di- and tri-thiol derivatives of chlorothalonil. The metabolic pathway involves conjugation with glutathione followed by metabolism to the cysteinyl conjugate which is then metabolised by lyase to the thiol metabolites. Examination of the kidney showed localisation of radiolabel in the kidney mitochondria. It was also shown that the di- thiol metabolite affects the respiration of kidney mitochondria. Both metabolites inhibit the transfer of reducing equivalents from succinate to coenzyme Q in the electron transport system. This is likely to result in reduced levels of ATP and a consequent impairment of the Na/K pump. In the rat, following oral dosing at 50 mg kg-1, 1.6% of the applied dose was excreted in urine as thiol metabolites. Following dermal dosing the rat excreted 0.07% of the applied dose as thiol metabolites. In Rhesus monkeys following oral dosing the tri-thiol metabolite was found in urine at up to 0.01% of the applied dose. Following dermal application less than 0.008% of the applied dose was excreted as thiols. Acute toxicology Chlorothalonil (technical) and formulations for which current approvals do not exist, but which contain up to 75% chlorothalonil, were of low acute oral and dermal toxicity. Technical chlorothalonil was toxic by inhalation. Chlorothalonil formulations ranged from toxic to unclassified by inhalation. A formulation containing 7% chlorothalonil was not classified as a skin irritant but was classified as irritating to eyes. There was some evidence that chlorothalonil was a skin irritant in humans and was possibly a skin sensitiser in animals when dissolved in an organic solvent. Following the ACPs consideration of the 1996 review of chlorothalonil in antifouling products, a data requirement for an adequate skin sensitisation study was set. Skin sensitisation studies submitted to address this data requirement were not sufficiently well conducted to draw conclusions on skin sensitisation. However, human data indicates that it has the potential to cause allergic dermatitis and there was also some, more limited evidence that it may cause contact urticaria. In the oral sub-acute studies, there was evidence of hyperplasia and hyperkeratosis of the forestomach although the lesion was reversed after a 13 w recovery period. The no observed effect level (NOEL) for hyperplasia was 3 mg kg-1 d-1. Hyperplasia of convoluted proximal kidney tubules was also noted. Unlike the forestomach hyperplasia there was only limited evidence of reversibility after a 13 w recovery period. There was evidence that with a dose of 175 mg kg-1 d-1, kidney lesions were produced by the fourth day of dosing. The NOEL for kidney effects was 3 mg kg-1 d-1 based on the incidence of intracytoplasmic inclusion bodies

following electron microscopic examination. However, a NOEL of 10 mg kg-1 d-1 was established based on conventional histological examination. Genotoxicity There was no evidence to suggest that chlorothalonil was mutagenic in vitro in bacterial reverse mutation assays (using rat liver or kidney S-9), chromosome aberration assays, mammalian cell gene mutation assays, DNA repair assays or unscheduled DNA synthesis assays. Chlorothalonil was not mutagenic in vivo in micronucleus assays, chromosome aberration assays or in a dominant lethal assay. Further data submitted for the review confirmed these earlier conclusions. As oxidative damage to DNA has also been recently demonstrated in vivo, this is again related to the cellular toxicity of chlorothalonil and there is no evidence that this causes long term effects in the intact animal. In addition, chlorothalonil did not cause cell transformations in cultivated rat embryo cells and did not bind to rat kidney DNA. The 17 metabolites and impurities of chlorothalonil were not mutagenic in a bacterial reverse mutation assay using rat kidney S-9. Carcinogenicity Chlorothalonil produced an increased incidence of hypertrophy/hyperplasia and adenomas/carcinomas of the proximal renal tubule epithelium in rats and mice. There was also evidence of renal tubular hypertrophy in 2 y dog studies. The critical NOELs for effects on the kidney were 2 mg kg-1 d-1 for renal tubule hyperplasia and 4 mg kg-1 d-1 for renal tumours from a 2 y rat study. Chlorothalonil also caused an increased incidence of hyperplasia and hyperkeratosis of the forestomach epithelium and an increased incidence of papillomas and carcinomas in the forestomach of rats and mice. The critical NOELs for effects in the forestomach were 1.86 mg kg-1 d-1 for hyperplasia from a 2 y mouse study and 4 mg kg-1 d-1 for forestomach tumours from a 2 y rat study. Reproductive Toxicity Chlorothalonil was not teratogenic in the rat or rabbit. There was no effect on reproductive performance in rat multigeneration studies. Human Data Studies on groups of workers and individuals occupationally exposed to chlorothalonil indicated that it had the potential to cause allergic dermatitis in humans, and there was also some, more limited evidence that it may cause contact urticaria. Risk Assessment for Human Health From the TER values, it is estimated that the application of chlorothalonil by spray is considered acceptable if operators wear air-fed RPE and coveralls of a contrasting colour to the product being applied, underneath a disposable coverall with hood, gloves and impervious footwear that protects the lower leg. To mitigate hand exposure, protective gloves should be replaced regularly, for example following antifoulant applications to a ship. Calculation of TERs suggests that this level of PPE would allow a large margin of safety vi

against systemic effects. Furthermore, the TER values estimate exposure for professionals (chandlers) by brush or roller would be considered acceptable if the appropriate PPE is worn. However, although the TER values for the application of chlorothalonil are acceptable, the potential exposure of amateurs to chlorothalonil by all methods is unacceptable based on the risk of skin sensitisation. No data are available for bystander exposure to antifouling products. However, there is minimal risk of skin sensitisation for bystanders. Environmental Fate and Behaviour Chlorothalonil was shown to be hydrolytically stable at pHs 7 and 5 (22 C) but at pH 9 a half-life of 38 d was calculated and the metabolites DS-19221 (54 %) and DAC-3701 (22 %) identified. Further investigations also revealed that DAC-3701 was extremely resistant to hydrolysis at all pHs tested. Chlorothalonil was shown to undergo aqueous photolysis and a half-life of 64.7 d was calculated. In aerobic sediment-water systems (fresh and seawater), chlorothalonil underwent rapid degradation with half-lives of < 2 h. The metabolites identified as a result of aquatic metabolism, in sediment-water systems, were not the same as those identified following chlorothalonil degradation in soil. Under aerobic aquatic conditions two major metabolites (SDS-67042 and SDS-67042 sulphoxide) and three minor metabolites were reported (SDS66432, SDS-13353 and SDS-3701). It should be noted therefore that the metabolites of concern in the aquatic environment have not been previously addressed due to the previous reviews of chlorothalonil concentrating solely on the agricultural use of chlorothalonil, where the contamination of the aquatic environment was considered unlikely. Chlorothalonil was reported to be of low mobility in various soil types. Two laboratory and one calculation method was submitted to address the leaching potential of chlorothalonil from antifouling paints. The laboratory studies reported comparable mean daily leaching rates of 3.06 and 3.18 g cm-2 d-1 from two antifouling paint formulations containing different levels of chlorothalonil. The theoretical study reported mean daily leaching rates of 0.99 and 1.70 g cm-2 d-1, which were notably less than those gleaned from the laboratory studies. Five studies investigated the bioaccumulation of chlorothalonil in two different fish species, bluegill sunfish and channel catfish. Both species were shown to reach a bioaccumulation equilibrium within 14 d and depurated at least 46 % of the accumulated ai in the same period of time under the selected test conditions. It should be noted that the BCFs recorded for both fish species studied were not calculated for chlorothalonil but for 14C-residues, and that subsequent characterisation studies revealed that chlorothalonil was not the major residue identified in water. The major 14C-activity in the bluegill sunfish tissues was characterised as unextracted, whereas the 14C-residue in the channel catfish was largely 14C-chlorothalonil.

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Ecotoxicology From the data submitted by the supporting company, chlorothalonil was shown to be of low toxicity to the freshwater algae Selenastrum subspicatus with an 5 d EC50 of 0.21 mg ai l-1. Acute exposure of invertebrates, Daphnia magna, Penaeus duorarum and Crassostrea virginiea, to chlorothalonil demonstrated significant toxicity but did not highlight major differences in sensitivities between freshwater and marine species with acute EC50 values of 97.20 (48 h), 165 (96 h) and 7.3 (96 h) g a.i l-1 respectively. Following chronic exposure to chlorothalonil, marine invertebrate demonstrated greater sensitivity than freshwater species, i.e. the Daphnia magna 21 d NOEC of 35 g ai l-1 was significantly higher than the 28 d NOEC of 0.83 g ai l-1 reported for Myopsis bahia. In freshwater fish, acute exposure to chlorothalonil resulted in high toxicity with similar LC50 values reported for the bluegill sunfish and the rainbow trout at measured concentrations of 35.10 and 33.25 g ai l-1 respectively. A 96 h LC50 of 32 g ai l-1 reported for the marine fish, sheepshead minnow, suggested that acute chlorothalonil toxicity was not significantly different to that reported for freshwater species. Under chronic conditions, a NOEC of 3 g ai l-1 was reported for the freshwater species, fathead minnow. The review of available literature where the sub-lethal effects of chlorothalonil against several aquatic species had been investigated, demonstrated that chlorothalonil could elicit significant toxic effects at quite low concentrations. In fish, at exposure levels of 1.4 g ai l-1 increased hepatic GSH activity predominated, and respiratory stress was noted at exposures exposed to 0.3 g ai l-1. However, in crustacea exposed to 0.3 g ai l-1 decreases in muscle or whole body AChE were significant. The suggestion is therefore that the differences in toxic profiles between fish and crustacea are not merely dose response but that the target of chlorothalonils action is different. Summary data suggested that chlorothalonil was of low toxicity to birds under the conditions tested. Environmental Risk Assessment The risk assessment has been concentrated on the marine environment since the data available are predominantly for the use of antifouling products (AFPs) in estuarine and coastal areas, although the risk to freshwater environments has not been precluded. However, the strategy for assessing risk to the marine environment is less well developed than for terrestrial or freshwater. Therefore, the risk assessment strategy adopted by HSE for the purposes of the current review has been presented in a supporting document Environmental Risk Assessment of Booster Biocides (ACP [2002]). This document presents a comprehensive and comparative risk assessment for all approved booster biocides and has been endorsed by an expert Ad-hoc Environmental Panel. Below are the main points of the risk assessment detailed in Environmental Risk Assessment of Booster Biocides concerned with the use of AFPs containing chlorothalonil, however, reference to the complete document is advised.

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Environmental Concentrations Usage information was obtained from a survey conducted by the Environment Agency (EA) in 1998. The EA survey demonstrated that chlorothalonil was not currently used to any significant level (< 3 % pleasure craft were treated with chlorothalonil). This was supported by 1998 monitoring data provided by CEFAS (Centre for Environment, Fisheries & Aquaculture Science) which demonstrated that chlorothalonil was not detected at any of the sampling sites. No sediment analysis was conducted for chlorothalonil. Since the current monitoring data could only ever represent the usage levels for 1998, predictions of maximum PECs were considered necessary as post approval usage cannot be controlled. Therefore, PEC data based on 100 % usage (all vessels treated) of chlorothalonil AFPs were calculated using a model developed as part of a HSE/EA commissioned research project. The model, REMA (Regulatory Environmental Modelling of Antifoulants), is a steady-state QWASI (quantitative water air sediment interface) model, designed to predict concentrations of biocides in both the water and sediment compartments of estuaries/marinas/harbours. The predicted water concentrations for marinas exceeded the limit of detection quoted by CEFAS (>1 ng l-1) regardless of use level, and in the estuary where the usage figure of 100 % was used. The mean PEC calculations for the sediment compartment were very low, and although it may be possible to detect these levels, at present there is no analytical information available to confirm this. Once released into the aquatic compartment, the chemical fate of the booster biocide will determine whether the toxic effect exerted is limited to the target organisms within a boundary layer of a painted surface or whether the ai persists and there is potential for exposure to non-target organisms. Therefore, selection of key non-target organisms and likely duration of exposure is essential, but this is somewhat reliant on the availability of acceptable data for representative marine species. Chronic data endpoints have been selected as more appropriate for the purpose of a marine risk assessment following the use of booster biocides. This is because the inputs of booster biocides into the marine environment as a result of leaching from multiple point sources (treated surfaces) will be a continuous process. Comparisons between marine and freshwater chronic toxicity data for booster biocides has not demonstrated any differences in sensitivities. Therefore, freshwater data have been accepted and the most sensitive species selected, regardless of test medium. The provision of assessment factors have been made in accordance with the guidance detailed in the European Risk Assessment Technical Guidance Document (EURATGD, 1996), and those previously accepted by the ACP. The most sensitive chronic endpoint available for chlorothalonil was a 28 d NOEC based on reproduction of 0.83 g ai l-1 for the marine mysid shrimp. The data set submitted for chlorothalonil included acute toxicity for both marine invertebrates and fish, and chronic studies for algae (freshwater), invertebrates (freshwater and marine) and fish (freshwater). Additional marine studies were also available in published studies. Therefore, an assessment factor of 10 was considered appropriate, giving a PNEC of 0.083 g ai l-1 for the mysid shrimp. Risk Quotient The PECwater:PNEC calculations derived from the predicted data based on 100 % usage suggest that for chlorothalonil, the only area where unacceptable risk would be likely to occur would be in marinas (open) with 100 % of the data exceeding a PEC:PNEC ratio of 1.

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These data were considered to demonstrate that the use of chlorothalonil in AFPs would not result in gross unacceptable effects. However, no refinement to the model parameters are considered necessary to address the risk from use on pleasure craft in the UK. Although outstanding data clarifications and metabolite data have been requested, refinement below the predicted levels was not considered possible considering the current limited information regarding appropriateness of short-term leaching rates to predict long-term steady-state effects. The PECsediment data derived by the REMA model suggest that the sediment compartment is not a concern for chlorothalonil. Therefore, no additional data to address chlorothalonils behaviour or toxicity in sediment have been requested. Efficacy Only limited laboratory screening data were available. It can be concluded from the screening studies, that toxicity of chlorothalonil was demonstrated against eight species of fungi, eight species of freshwater algae, and nine species of marine algae. Chlorothalonil appeared to be generally more active against freshwater algae than TBTO. MICs for fungi and marine algae were in the range of 0.1 - > 1000 mg l-1 chlorothalonil. Overall Proposals and Data Requirements The following recommendations and data requirements were agreed by Ministers:
1. All amateur uses of antifouling products containing chlorothalonil are to be revoked because information from humans indicates that the risk of skin sensitisation is unacceptable.

Approvals for the professional use of antifouling products containing chlorothalonil at a maximum concentration of 5 % w/w may be continued for professional use by brush, roller and spray, subject to the data requirements and conditions detailed below:
2.

Physical Chemistry For technical chlorothalonil: 1 All data to be submitted within 1 year unless otherwise specified. Manufacturers must demonstrate to HSE how they are implementing the requirements within 6 months. From Company No. 1: a) b) c) d) e) f) g) Address of manufacturing site. Results from typical batch analyses conducted on technical chlorothalonil, with associated test reports (e.g. chromatographic traces should be included). A minimum - maximum range of purity for the active ingredient should also be stated. UV and IR traces to support the results provided. Further spectral data to include 13C NMR spectra and MS (traces should be provided). Surface tension including full test report. Full test reports for: melting point, boiling point, relative density, vapour pressure,solubility in water, partition coefficient, and explosive properties. Protocol reports, plus typical experimental results including example traces and validation data, for the analytical methods provided for the determination of x

h)

chlorothalonil in the technical material and water. The analytical method used to determine chlorothalonil in water should be able to determine the active substance to 0.1 g l-1. Analytical methods to determine chlorothalonil in antifouling products. Protocol reports, plus typical experimental results including example traces and validation data should be included.

From Company No. 2: a) b) c) d) e) f) g) h) BSI name, IUPAC name, other names, codes and synonyms. Identification numbers including CAS and EINECS numbers. Molecular mass, molecular formula, structural formula. Site of manufacture (including company name and address). Technical specification including min. - max. purity of active substance expressed in % w/w or g kg -1 and identification and quantification of significant impurities and those of toxicological concern expressed as max. % w/w or g kg -1. Results from typical batch analyses conducted on technical chlorothalonil, with associated test reports (e.g. chromatographic traces should be included). Spectral data, to include UV, IR, 13C NMR spectra and MS (traces should be provided). The following endpoints (including full test reports): melting point, boiling point, relative density, surface tension, vapour pressure, solubility in water, partition coefficient log Pow, flammability*, oxidising* and explosive properties*. * As an alternative, reasoned cases may be provided. Analytical methods for determination of chlorothalonil and impurities in technical material to support the technical specification data requirement in e). Protocol reports, plus typical experimental results including example traces and validation data should be included. Analytical method to determine chlorothalonil in water and in antifouling products formulations. The analytical method used to determine chlorothalonil in water should be able to determine the active substance to 0.1 g l-1.

i)

j)

From all Approval Holders: Storage stability studies on representative current products, under ambient conditions in the product packaging. If the approval holder undertakes to conduct new studies, HSE will require evidence that such work is underway after a period of 6 months. New long term storage stability studies should be provided within 3 years. Protocols are to be agreed with HSE. 1. Human Health Data are to be submitted within 1 year. Approval holders must demonstrate to HSE how they are implementing the requirements within 6 months. a) A study of the dermal penetration of chlorothalonil from formulations representative of approved antifouling products, to be submitted within 1 year. This study may be carried out in vitro or in vivo. The protocol should be discussed with HSE prior to commencement.

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b)

Professional operators (sprayers) exposed to antifouling products containing chlorothalonil must wear RPE. Appropriate RPE includes air-fed respiratory equipment with combined protective helmet and visor to protect the skin of the head and neck. Impairment of vision should be avoided. For workers other than sprayers in the vicinity of the spray plume, RPE of an equivalent standard to FFP3 should be worn. For other workers, the need for RPE should be informed by a COSHH assessment. Unprotected persons should be kept out of treatment areas to minimise the risks posed by exposure to this compound. All professional operators exposed to antifouling products containing chlorothalonil should wear a disposable coverall with hood (providing head protection) and a second overall beneath this coverall of a contrasting colour to the antifouling product being applied. All bare skin should be covered. The disposable coverall should normally be used for no more than one spraying session. The second overall should be changed regularly and whenever product break-through has been detected. Professional operators working with antifouling products containing chlorothalonil should wear impermeable gloves of a type recommended by the antifouling manufacturer as suitable for use with the formulation. These gloves should be changed regularly, e.g. after one or two days use. Operators should wear impermeable (and non-slip) footwear that protects the lower leg. The following approval conditions are to appear on products' Notice of Approval and Schedules and they should be reflected on product labels using the following precautionary phrases: WEAR SUITABLE PROTECTIVE CLOTHING (COVERALLS OF A CONTRASTING COLOUR TO THE PRODUCT BEING APPLIED, BENEATH A DISPOSABLE COVERALL WITH HOOD), SUITABLE GLOVES, AND IMPERVIOUS FOOTWEAR THAT PROTECTS THE LOWER LEG. DISPOSE OF PROTECTIVE GLOVES AFTER USE If the product is to be applied by spray: UNPROTECTED PERSONS SHOULD BE KEPT AWAY FROM TREATED AREAS DO NOT BREATHE SPRAY MIST WEAR SUITABLE RESPIRATORY EQUIPMENT SUCH AS AIR-FED RESPIRATORY EQUIPMENT WITH COMBINED PROTECTIVE HELMET AND VISOR WHEN SPRAYING. WEAR SUITABLE RESPIRATORY EQUIPMENT SUCH AS FFP3 OR AN EQUIVALENT STANDARD WHEN WORKING IN THE VICINITY OF THE SPRAY PLUME

c)

d)

e)

f)

xii

Environment The following data requirements are to be submitted within a total of 3 years, with shorter timetables for some requirements as outlined. Approval holders must demonstrate to HSE how they are implementing the data requirements within 6 months. a) Inhibition of microbial respiration in sewage sludge (core requirement) to be submitted within 12 months. b) Fish early-life stage test (sheepshead minnow preferable species) to be submitted within 12 months. c) Clarification that the leaching rate of the parent ai used for the risk assessment is representative of current products. This information is to be submitted within 12 months. (Repeated REMA calculations will be carried out by HSE with any new data supplied.)

d) Should the leaching data submitted as part of requirement c) fail to reduce the predicted environmental concentrations of chlorothalonil to acceptable levels (assuming 100 % use on pleasure vessels) a monitoring programme will be necessary. In the first instance, HSE should liaise with DETR, EA and English Nature and investigate the potential for further monitoring via the National Marine Monitoring Programme or other organisations. e) Confirmation that chlorothalonil significantly degrades in marine sediment-water systems, and identification of any subsequent persistent metabolites to be submitted within 12 months. Once the significance and persistence of the metabolites have been established additional ecotoxicology, fate and behaviour data may be required in order to permit a risk assessment to the same level as that of the parent compound. The specific data required should be discussed with HSE on completion of data requirement e) and all subsequent data requirements submitted within a further 2 years.

f)

g) A study to address bioaccumulation of chlorothalonil in shellfish, in order to address the potential risks to consumers via the food chain, should be submitted within 2 years. Efficacy The following efficacy data are to be provided within 3 years. If approval holders undertake to conduct new studies, they must demonstrate to HSE how they are implementing the data requirements within 6 months. To support the continued use of antifouling products containing chlorothalonil, raft test or alternatively field trial/in service monitoring data are required. These data should cover: a) The efficacy of TBT-free SPC coating antifouling formulations that contain chlorothalonil. xiii

The full nature and extent of data required is to be outlined to individual approval holders, as appropriate, by the HSE. In all cases, data should be provided by approval holders in support of the coating type(s) that best befits their products. If a product is assigned to a different and more accurate coating type by an approval holder, and is, as a consequence, not supported by existing information, appropriate efficacy data must be submitted. Alternatively for a), justification or evidence based on data may be provided through the submission of bridging studies.

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1 INTRODUCTION AND REGISTRATION HISTORY

1.1 INTRODUCTION AND BACKGROUND TO REVIEW
Chlorothalonil is used as a fungicide in agriculture and is also used in approved antifoulant products. This active ingredient was first considered by the Scientific Subcommittee on Pesticides (SCP) in 1967 where it gained provisional approval for use on non-edible crops, potatoes and culinary apples. 1.1.1 EARLIER REVIEW OF AGRICULTURAL USE

A special review of the agricultural uses of chlorothalonil was initiated by MAFF (now DEFRA) in October 1990. This was prompted by concerns over reports from Europe indicating that chlorothalonil may cause forestomach papillomas, squamous cell carcinomas and renal tubule adenomas and carcinomas in rats and mice. The SCP deferred recommendations until the registrant had produced relevant data addressing the toxicological and operator exposure concerns. The concerns relating to possible carcinogenicity were addressed in a Position Paper which was discussed by the SCP in March 1993. This Position Paper addressed only chlorothalonil toxicokinetics, repeat dose, genotoxicity, carcinogenicity, teratology and operator exposure studies. It was concluded that chlorothalonil was not a genotoxic carcinogen but the proposed mechanism was uncertain. The ACP considered chlorothalonil in January 1994 (ACP, 1994) and agreed an AOEL of 0.03 mg kg-1 d-1, based on a NOEL of 3 mg kg-1 d-1 with a 100 fold margin of safety. The NOEL was derived from a 13 week study in Sprague Dawley rats and was based on hyperplasia and hyperkeratosis of the forestomach at the next highest dose of 10 mg-1 kg-1 d-1. A figure of 0.5% was agreed for percutaneous absorption. Following the agricultural review, a partial review of the use of chlorothalonil in antifouling products and aquaculture was present to the SCP in April 1994. This paper initiated the review of the non agricultural uses of chlorothalonil as a follow on from the review of its agricultural uses. The Committee determined that no decision could be made on the review until usage data had been provided. The review was represented to the SCP in October 1995 and ACP in January 1996 and included limited usage data together with some limited mammalian and environmental toxicity data in areas not already covered by the review of the agricultural uses. The toxicity data seen previously is summarised in Section 3. Ministers agreed to the continued approval of chlorothalonil containing products subject to the submission of data to address skin sensitisation, operator exposure, residues in fish, environmental fate and behaviour and environmental toxicity. 1.1.2 REVIEW OF ANTIFOULANT USE

In 1995, the ACP considered reviews of the triorganotin compounds (TOTs) in antifouling products and copper compounds. It concluded that the risk from the use of TOTs was unacceptable but that the use of copper antifouling products (AFPs) was acceptable. However, the ACP recommended that a high priority should be placed on reviewing the organic 'booster biocides', since restrictions on the use of TOT-containing products could lead 1

to an increased environmental loading of these compounds. Booster biocides is a generic term given to a group of compounds normally found in addition to copper and occasionally TOT, in AFPs. The purpose of these compounds is to enhance the products efficacy against a broader spectrum of fouling organisms than that achieved with copper alone. Booster biocides are either organic or organo-metallic compounds which have fungicidal, herbicidal or anti-microbial actions. The ACP considered the full review of chlorothalonil at its 278th meeting in September 2000. It recommended that provisional approval for the professional use by brush, roller and spray, of antifouling products containing chlorothalonil at a maximum concentration of 5 % w/w may be allowed to continue; subject to the data requirements and conditions (detailed in Chapter 9). In addition, it recommended that all amateur uses of antifouling products containing chlorothalonil should be revoked because information from humans indicated that the risk of skin sensitisation was unacceptable. Ministers accepted these recommendations. This paper presents an updated review of the physical chemistry, mammalian toxicokinetics and toxicology, environmental and efficacy of chlorothalonil. It comprises a summary of data presented in earlier reviews together with an evaluation of new data submitted in response to earlier data requirements. An update of the published literature is also included. The assessment of risks to users and bystanders has been revised to take into account all factors affecting exposure and increase the clarity and transparency.

1.2 REGISTRATION HISTORY OF ANTIFOULING PRODUCTS

1.2.1 BACKGROUND

In July 1987, antifouling products were brought under The Control of Pesticides Regulations (COPR), 1986. As a result of discussions between Government Departments and the antifouling product industry, it was agreed that these products would not be subject to the same registration procedures as other non-agricultural pesticides until their active ingredients were fully reviewed. Companies producing antifouling products had been invited in April 1987 to inform the Health and Safety Executive (HSE) of all active ingredients used in the formulations that they wished to be included under the Regulations - although many of these had not been previously evaluated by the ACP, there was no requirement at that time to submit any safety or other data to support their applications. In addition, antifouling companies were permitted to seek approval for products where, unlike other pesticide products, the active ingredient(s) was specified in terms of a percentage range instead of a fixed value and that there were no restrictions placed upon the number of active ingredients permitted in each antifouling product. It had been further agreed, and published in The Edinburgh Gazette of Friday 26 June 1987 to this effect, that antifouling products should be classified and labelled in accordance with the 77/728/EEC Council Directive relating to classification, packaging and labelling of paints, varnishes and inks (European legislation which has subsequently been superseded by the 88/379/EC Council Directive relating to the classification, packaging and labelling of dangerous preparations). As an outcome of this decision with respect to labelling of

antifouling products, it was only necessary for the names of certain dangerous substances to appear on product labels rather than each active ingredient and its percentage in w/w. The first approvals of antifouling products containing 2,3,5,6-tetrachloroisophthalonitrile (known more commonly under its BSI common name of chlorothalonil), granted on 29 June 1987, were given under these transitional arrangements. In 1991, proposals attempting to terminate the transitional arrangements granted to antifouling products was presented to the Committees, so that all pesticide products would be dealt in a similar fashion. However, it was considered prudent to leave these arrangements in place until a full review of all active ingredients found in antifouling products had taken place. 1.2.2 PROPOSED INTERNATIONAL BAN ON ORGANOTINS

The recently-agreed position of the International Maritime Organisation (IMO), has made it likely that there will be an early world wide ban on the use of organotin compounds in antifouling systems. On 22 November 1999 the IMO General Assembly passed a resolution, put forward by the Marine Environment Protection Committee (MEPC), that that the MEPC should develop a global, legally binding, instrument to address the harmful effects of antifouling systems used on ships. The assembly also agreed that the instrument should ensure a global prohibition of the application of organotin compounds by 1 January 2003 and the prohibition of its presence on ships by 2008. At present there is no guidance on the definition of 'organotin compounds' and whether this covers both copolymer and free-association systems. A working group within MEPC has started to draft a free standing international convention to regulate shipboard antifouling systems that have adverse effects on the marine environment. This will include an annex of systems subject to specific controls, including a ban on their use. IMO Council agreed that there should be a Diplomatic Conference on the convention in 2001. It should be noted that any ban on the use of organotins in antifouling products could result in increased use of any or all of the booster biocides. 1.2.3 CURRENTLY APPROVED ANTIFOULING PRODUCTS

In September 2000, there were 270 approved antifouling products that could legally be marketed in the UK and, of these, 4 contained chlorothalonil. These 4 product approvals are held by 2 approval holders. The distribution of chlorothalonil antifouling products by active ingredient is shown in Table 1.1. Table 1.1: The distribution of chlorothalonil antifouling products by active ingredient ACTIVE INGREDIENT (S) Chlorothalonil/Cu2O/Diuron Chlorothalonil/Cu2O/Diuron/Irgarol 1051 Chlorothalonil/Cu2O/RH-287 No 2 1 1

Both approval holders have submitted information on maximum dry-docking intervals (36 months) for all of their products. Applying antifoulant requires 2 to 4 persons per spray position. The three identified tasks are: spraying - the sprayer; mixing and loading - the pot-man (who prepares the antifoulant and ensures its supply to the high pressure pump), ancillary - the rein or tender men, who attend to keeping paint lines free and may also manoeuvre the mobile access platform (cherry-picker).

Chlorothalonil is an active ingredient permitted in approved professional and amateur antifouling products for use on vessels of any size plus structures below the waterline such as oil platforms, jetties, navigation buoys or piers. It cannot be used in aquaculture (on apparatus or equipment used in the cultivation of fish and shellfish plus fishing nets, lobster pots etc). The maximum concentrations of chlorothalonil permitted as an active ingredient within approved formulations are:Products applied to vessels by amateurs : Products applied to vessels by professionals : 5.0 % w/w 5.0 % w/w

At present, all 4 of the currently approved antifouling products containing chlorothalonil are intended for amateur and professional use. Although products may contain up to 5.0 % w/w of the compound, the current maximum level in existing products is only 3.0 % w/w. Application rates recommended by companies for products containing chlorothalonil vary depending upon expected service life of the antifouling coating. However, all current products are applied at a rate of 1 litre per 5 m2 with recommended repainting after 36 months. Acceptable application methods for applying approved products containing chlorothalonil to vessels/structures are by brush, roller and airless or conventional spray. 1.2.4 CLASSIFICATION AND LABELLING OF ANTIFOULING PRODUCTS CONTAINING UP TO 5 % w/w OF CHLOROTHALONIL

1.2.4.1 Background Under transitional agreements reached between Government Ministers and industry in 1987, antifouling products are classified and labelled as if they were paints and must comply with current EC legislation, namely Council Directive 88/379/EC relating to the classification, packaging and labelling of dangerous preparations (which is currently implemented in the UK as The Chemicals (Hazard Information and Packaging for Supply) Regulations 1994). This Dangerous Preparations Directive (and its subsequent adaptations) allows EC classifications assigned to each substance within a preparation to be pooled together in order to determine overall EC Classifications for the product, by use of an agreed set of calculations and assumptions as laid out in an annex to this Directive.

The EC classification(s) for each substance used in this determination of product classification must be taken, whenever possible, from a European inventory - Annex I to Council Directive 67/548/EC relating to the classification, packaging and labelling of dangerous substances. This Annex contains EC classifications for a number of compounds/elements which have been agreed by EC Member States after consideration of all available toxicological and environmental data. Therefore, where substances have already been considered by EC Member States, decisions taken on their classification(s) will override any such decisions taken at a national level. If a substance does not have an entry on Annex I of Directive 67/548/EEC, then data provided on the manufacturers Material Safety Data Sheet for that substance should be used in determining overall classification. However, it should be pointed out that should actual formulation data be available, then this will override any determination of product classification which has been derived from the classifications of its component substances. 1.2.4.2 Classification / Labelling Under UK Legislation Chlorothalonil itself has an entry on Annex I to Directive 67/548/EEC (reproduced in the UK as various editions of The HSC Approved Supply List) stating that it is classified as :CARCINOGENIC (Xn) : (Category 3) R40 : POSSIBLE RISK OF IRREVERSIBLE EFFECTS.

At present, Council Directive 88/379/EEC is implemented as The Chemicals (Hazard Information and Packaging for Supply) Regulations 1994 - CHIP 2. Recent adaptations to the Directive have been introduced into the UK as amendments to CHIP 2, the latest set being presented as CHIP 99. Currently no environmental labelling is required under CHIP. However, it is to be introduced in the near future and antifouling products will be labelled in line with the consequent requirements. Therefore, antifouling products containing 1.0 - 5.0 % w/w of chlorothalonil would attract the following classification under CHIP 99 :EC Classification : Hazard symbol : Risk phrase : CARCINOGENIC (Category 3) ; HARMFUL (ST ANDREWS CROSS) ; R40 : POSSIBLE RISK OF IRREVERSIBLE EFFECTS. R50: VERY TOXIC TO AQUATIC ORGANISMS R53: MAY CAUSE LONG TERM ADVERSE EFFECTS IN THE AQUATIC ENVIRONMENT

Dangerous for the Environment (N)

1.2.5 TYPICAL FRAME FORMULATION OF A YACHT ANTIFOULING PAINT A typical yacht paint consists of biocide and pigment dispersed in a resinous binder, reduced to an acceptable application viscosity with solvent. Additives are incorporated to modify paint film properties, application or storage characteristics.

2 PHYSICAL CHEMISTRY OF CHLOROTHALONIL

The data presented below is for technical chlorothalonil manufactured by Company No. 1. Currently there are two known manufacturers of chlorothalonil, Company No.1 and Company No.2 used in antifouling products. Previous data seen in ACP (1994) for the active substance are shown in italics. Identity, physico-chemical properties and analytical methods for chlorothalonil have been summarised. No test reports for the physico-chemical properties have been submitted.

2.1 IDENTITY OF THE ACTIVE SUBSTANCE

BSI Common Name: Chlorothalonil IUPAC Names: Tetrachloroisophthalonitrile or tetrachlorodicyanobenzene Other synonyms or codes: TPN(JMAF); EU number: 608-014-004 CAS number : 1897-45-6 EINECS number: 217-588-1 Molecular formula: C8Cl4N2 Molecular mass: 265.9 Structural formula:
CN Cl Cl

Cl Cl

CN

Percentage Purity of technical active ingredient minimum purity: 985 g kg-1 (Range: Not given) Spectral data: The endpoints provided from spectral data (UV and IR) are broadly consistent with the chemical structure of the compound. No traces were supplied. (Unpublished, 1999) DATA REQUIREMENTS: For technical chlorothalonil: From Company No. 1: 1. Address of manufacturing site. 2. Results from typical batch analyses conducted on technical chlorothalonil, with associated test reports (e.g. chromatographic traces should be included). A minimum - maximum range of purity for the active ingredient should also be stated. 3. UV and IR traces to support the results provided.

4. Further spectral data to include 13C NMR spectra and MS (traces should be provided). From Company No. 2: 1. BSI name, IUPAC name, other names, codes and synonyms. 2. Identification numbers including CAS and EINECS numbers. 3. Molecular mass, molecular formula, structural formula. 4. Site of Manufacture (including company name and address). 5. Technical specification including min. - max. purity of active substance expressed in %w/w or g kg -1 and identification and quantification of significant impurities and those of toxicological concern expressed as max. % w/w or g kg -1. 6. Results from typical batch analyses conducted on technical chlorothalonil, with associated test reports (e.g. chromatographic traces should be included). 7. Spectral data, to include UV, IR, 13C NMR spectra and MS (traces should be provided).

2.2 PHYSICO-CHEMICAL PROPERTIES

Physical state at 25 oC and 101.3 KPa: Melting point: White solid (Unpublished, 1999) Colourless odourless crystals 252.5 - 254.5 oC (pure chlorothalonil) (Unpublished, 1999) 250 - 251 oC ~350 oC (Unpublished, 1999) 350 oC 1.73 (Unpublished, 1999) Not provided 0.22 mPa (Unpublished, 1999) 1.3 Pa 5.9 mPa 0.54 mg l-1 (Unpublished, 1999) 0.6 mg kg -1 Partition Coefficient: log Pow at 22 oC: Flammability: Oxidising properties: 2.93 (Unpublished, 1999) 4.0 (temperature not stated) Not flammable (Unpublished, 1999) Not oxidising (Unpublished, 1999)

Boiling point: Relative density D420: Surface tension at 25 oC: Vapour pressure at 25 oC: at 40 oC: at 45 C: Solubility in water at 25 oC:
o

Explosive properties: DATA REQUIREMENTS: For technical chlorothalonil: From Company No. 1: 1. Surface tension including full test report.

Not explosive (Unpublished, 1999)

2. Full test reports for: melting point, boiling point, relative density, vapour pressure, solubility in water, partition coefficient, and explosive properties. From Company No. 2: The following endpoints (including full test reports): melting point, boiling point, relative density, surface tension, vapour pressure, solubility in water, partition coefficient log Pow, flammability*, oxidising* and explosive properties*. * As an alternative reasoned cases may be provided.

2.3 STORAGE STABILITY

DATA REQUIREMENTS: From Company No. 1 and Company No. 2: 2 year storage stability studies on representative formulations at ambient temperature.

2.4 ANALYTICAL METHODS

The following methods for the determination of chlorothalonil were submitted. 1a. Analytical method for the determination of chlorothalonil in the technical material by GC-FID. (Unpublished, 1999) 1b. Analytical method for the determination of chlorothalonil in the technical material by GC-FID. 2. Analytical method for the determination of impurities in the technical material by GC-FID. (Unpublished, 1999) 3. Analytical method for the determination of the active ingredient in aqueous solutions by GC-ECD. (Unpublished, 1999) DATA REQUIREMENTS: From Company No. 1: 1. Protocol reports, plus typical experimental results including example traces and validation data, for the analytical methods provided for the determination of chlorothalonil in the technical material and water*. Analytical methods to determine chlorothalonil in antifouling products. Protocol reports, plus typical experimental results including example traces and validation data should be included.

2.

From Company No. 2: 1. Analytical methods for determination if chlorothalonil and impurities in technical material to support the technical specification data requirement in 2.1. Protocol reports, plus typical experimental results including example traces and validation data should be included. Analytical method to determine chlorothalonil in water* and in antifouling products formulations.

2.

* The analytical method used to determine chlorothalonil in water should be able to determine the active substance to 0.1 g l-1. Manufacturers must demonstrate to HSE how they are implementing the data requirements within six months.

10

MAMMALIAN TOXICOKINETICS AND TOXICOLOGY

3.1 BACKGROUND
This chapter comprises a summary of studies previously evaluated by the ACP for a review of the agricultural and horticultural uses of chlorothalonil (ACP, 1994). In addition, the limited mammalian toxicity data included in the review of chlorothalonil use in antifouling and aquaculture products presented to the ACP in 1996, has been summarised in this chapter. All summaries of data previously seen by Committees are given in italics. In addition an updated literature search was carried out; relevant studies have been evaluated in full and are presented in the appropriate section.

3.2 MAMMALIAN TOXICOKINETICS

Following oral dosing chlorothalonil was absorbed and rapidly eliminated by the rat (t1/2 6 7 h). In an in vitro study it was shown that chlorothalonil was only absorbed from the duodenum as metabolites. A rat dermal absorption study showed that 6 % of a 5 mg kg-1 dose was absorbed in 8 h. In an in vitro absorption study with human epidermis, the rate of absorption was 0.034 g cm-2 h-1 (ACP, 1994). Highest tissue concentrations of chlorothalonil were found in the liver and kidneys; maximum levels were reached 24 h post dose. A more detailed study, showed localisation of the radiolabel in the kidney mitochondria. Metabolism studies showed that the main metabolites formed were the mono-, di- and tri-thiol derivatives of chlorothalonil. The metabolic pathway involves conjugation with glutathione followed by metabolism to the cysteinyl conjugate which is then metabolised by lyase to the thiol metabolites. In in vitro studies, it was shown that the di-thiol metabolite affects the respiration of kidney mitochondria and that both di and tri metabolites inhibit the transfer of reducing equivalents from succinate to coenzyme Q in the electron transport system. This latter effect is likely to result in reduced levels of ATP and a consequent impairment of the Na/K pump (ACP, 1994). In rats, excretion in the faeces (82 - 92%) accounted for a greater proportion of the dose than excretion in the urine (5 - 12%). Most of the radiolabel was excreted within 24 - 48 h (urine) and 48 - 72 h (faeces). In a study to investigate biliary excretion, a large fraction of the dose was not absorbed since recovery of radiolabel from the faeces and g.i. tract accounted for 58 % of the dose in males and 64 % of the dose in females. A greater proportion of radiolabel was excreted in the bile (21 % in males and 17 % in females) compared to the urine (9 % in males and 12 % in females). In the rat, following oral dosing 21 % of the total urine radiolabel was excreted as thiol metabolites, within 24 h. Following dermal application, the maximum amount of thiol metabolite excreted, within 24 h, was 2.7 % of the total urine radiolabel (ACP, 1994). In Rhesus monkeys following oral dosing, the majority of the radiolabel was excreted in the faeces within 96 h with faecal excretion accounting for 52 - 92 % of the applied dose. Excretion in the urine accounted for 2 - 4 % and was essentially complete within 48 h. Following dermal application, excretion of the radiolabel in the urine and faeces was low 11

(0.6 - 1.4 % of the applied dose), as were tissue residues (0.08 - 0.64 % of the applied dose) (ACP, 1994). Two additional studies of chlorothalonil in animal systems have been published since 1995. The studies assessed in ACP (1994) indicated that 3-5% of an oral dose of chlorothalonil was absorbed, as glutathione conjugates, and a comparison between germ-free (GF) and conventional (CV) rats indicated that gut flora played an important role in absorption. New work on 14C-labelled chlorothalonil in an ex vivo everted rat gut system has confirmed the low level of absorption (less than 1% under the test conditions). Although slightly more unchanged chlorothalonil was absorbed across the gut from GF animals, only the CV gut was able to metabolise chlorothalonil; mono-, di-, and trimethylthiolated metabolites were identified. In addition, the amount of radioactivity retained in duodenal tissue was significantly greater in the CV rat preparation than from GF animals, possibly due to morphological modification or reduced biotransforming abilities of GF animals. This study does not alter the overall understanding of the toxicokinetics of chlorothalonil gained from the papers assessed previously (Hillenweck et al, 1998). A study of the cytotoxic effect of chlorothalonil on isolated rat hepatocytes and cell fractions confirms the previous view that the metabolic pathway involves conjugation with glutathione. The authors also found that chlorothalonil stimulated lipid peroxidation, possibly related to thiol oxidation, though this did not seem to be a cause of cell death. This paper does not add any substantial new information to the picture given in ACP (1994) (Yamano and Morita, 1995).

3.3 MAMMALIAN TOXICOLOGY

3.3.1 ACUTE TOXICITY Chlorothalonil (technical) and formulations for which current approvals do not exist, but which contain up to 75% chlorothalonil, were of low acute oral and dermal toxicity (Unpublished, 1985; 1987; 1987a). Technical chlorothalonil was toxic by inhalation (Unpublished, 1985). Chlorothalonil formulations ranged from toxic to unclassified by inhalation (Unpublished, 1985). A formulation containing 7% chlorothalonil was not classified as a skin irritant but was classified as irritating to eyes (Unpublished, 1987b; 1987c). There was some evidence that chlorothalonil was a skin irritant in humans (Unpublished, 1985). 3.3.2 SKIN SENSITISATION There was some evidence that chlorothalonil was possibly a skin sensitiser in animals when dissolved in an organic solvent but not when applied as a powder moistened with saline (Unpublished, 1985). Following the ACPs consideration of the 1996 review of chlorothalonil in antifouling products, a data requirement for an adequate skin sensitisation study was set. This requirement has been addressed and is evaluated as follows:

12

Groups of 20 test and 10 control Hartley guinea-pigs were treated with Demidekk Neutral Base A, Demidekk Neutral Base B or Demidekk Neutral Base C dissolved in deionised water, according to the three induction protocol of Buehler. The chlorothalonil contents or purity of the test formulations were not stated and could not be ascertained by the approval holder. In an extensive pilot study (12 animals/formulation), 30 - 100% solutions of the 3 formulations produced only very slight, usually non-confluent, erythema at all concentrations. In the main study, the neat formulations were used for induction and 30% solutions for challenge. Slightto-moderate erythema was noted at induction. At challenge, the responses in both treated animals and naive controls were limited to very slight erythema. Therefore, under the conditions of this study, Demidekk Neutral Base A, Demidekk Neutral Base B and Demidekk Neutral Base C did not show any evidence of skin sensitising potential. A positive control group, treated with DNCB, gave appropriate responses. However, since the compositions and chlorothalonil contents of the Demidekk formulations are uncertain, it is not possible to conclude that this study has adequately measured the ability of chlorothalonil to cause skin sensitisation (Unpublished, 1988). In a briefly-reported maximisation study, a group of 10 Hartley guinea pigs was exposed to Daconil, a preparation containing 75% chlorothalonil. At induction, 5% Daconil in distilled water was used intradermally, and 25% topically. Subirritant concentrations of 5% and 0.5% were chosen for challenge, apparently based on reactions in naive animals. At challenge, 90% of animals reacted to both concentrations, possibly providing evidence of a sensitising effect. However, no concurrent controls were used in this study, so that it is not possible to exclude an irritant reaction. In addition, although this study raises concerns that chlorothalonil may be a skin sensitiser, Daconil contains other components that may have contributed to the production of the skin reactions observed. Overall, these findings do not allow any firm conclusions to be made regarding the skin sensitising potential of chlorothalonil (Matsushita and Aoyama, 1981). 3.3.2.1 Summary The information available from animal studies is of very limited use. A poorly-conducted guinea pig maximisation test carried out on a formulation containing 75% chlorothalonil did not provide any meaningful conclusions, while Buehler tests on 3 formulations containing uncertain amounts of chlorothalonil were negative. Although a very briefly reported skin sensitisation study considered by the ACP in 1996 was apparently positive when chlorothalonil was administered dissolved in organic solvents, no firm conclusion on the ability of chlorothalonil to induce skin sensitisation in guinea pigs can be made in the absence of a well-conducted and properly-reported skin sensitisation test on the substance itself. 3.3.3 SUBACUTE STUDIES In the oral sub-acute studies, there was evidence of hyperplasia and hyperkeratosis of the forestomach although the lesion was reversed after a 13 w recovery period. The no observed effect level (NOEL) for hyperplasia was 3 mg kg-1 d-1. Hyperplasia of convoluted proximal kidney tubules was also noted. Unlike the forestomach hyperplasia there was only limited evidence of reversibility after a 13 w recovery period. There was evidence that with a dose of 175 mg kg-1 d-1, kidney lesions were produced by the fourth day of dosing. The NOEL for kidney effects was 3 mg kg-1 d-1 based on the incidence of intracytoplasmic inclusion bodies following electron microscopic examination. However, a NOEL of 10 mg kg-1 d-1 was established based on conventional histological examination (ACP, 1994). 13

3.3.4 GENOTOXICITY There was no evidence to suggest that chlorothalonil was mutagenic in vitro in bacterial reverse mutation assays (using rat liver or kidney S-9), chromosome aberration assays, mammalian cell gene mutation assays, DNA repair assays or unscheduled DNA synthesis assays. Chlorothalonil was not mutagenic in vivo in micronucleus assays, chromosome aberration assays or in a dominant lethal assay. In addition, chlorothalonil did not cause cell transformations in cultivated rat embryo cells and did not bind to rat kidney DNA (ACP, 1994). The 17 metabolites and impurities of chlorothalonil were not mutagenic in a bacterial reverse mutation assay using rat kidney S-9 (ACP, 1994). An updated search of the published literature has revealed further studies that have investigated the genotoxic potential of chlorothalonil, and these are described below. 3.3.4.1 In Vitro Studies in Mammalian Cells 3.3.4.1.1 Metaphase Analysis Chinese hamster ovary cells were incubated for either 2 hours (with Aroclor-induced rat liver S9) or 21.5 hours (without S9) with concentrations of chlorothalonil that apparently extended into the cytotoxic range although details of this were not presented. The harvest time was 21.5 hours after the start of treatment. Only one experiment was carried out, and there was apparently no replicate sampling. In the test without S9 (concentrations 0.5 to 0.7 g ml-1), there were random increases in the percentage of cells with aberrations (excluding gaps) but the absence of a dose-response indicated that this was not a true positive result. With S9 however (concentrations 2 to 3 g ml-1) there were significant increases (26% cells with aberrations [excluding gaps] at 3 g ml-1 compared to zero percent in controls) with some evidence of a dose response, but with a rapid rise and subsequent plateau. Positive controls behaved appropriately (Galloway et al, 1987). The ability of chlorothalonil to induce chromosome aberrations in Chinese hamster ovary K1 cells has been investigated only in the absence of an exogenous source of metabolic activation. The cells were exposed for 4 hours, followed by fixation at 24 hours. Although there were negative controls, duplicate determinations and two independent experiments, there were no positive controls and apparently only two dose levels tested (0.25 and 0.5 M). The top dose was chosen to cause 20-25% cytotoxicity in a test for oxidative activity that measured delayed effects. The same size increase in the percentage of aberrant cells was seen at both test concentrations compared to negative controls (15% compared to 4%) but with no evidence of an overall dose-response (Vigreux et al, 1998). These two studies indicate that although increases in aberrations have been seen both in the presence and absence of metabolic activation, the profile of the responses, especially the lack of a clear dose-response, reflected damage due to cytotoxicity rather than clastogenicity.

14

In a poor quality study, human lymphocytes were incubated with a mixture of 15 pesticides, including 13.1% chlorothalonil, in an assay designed to look at cytogenetic effects. Few experimental details were provided. A positive result was obtained that was traced to the Benomyl in the mixture. The nature of this study makes it of very limited value in assessing the genotoxicity of chlorothalonil (Dolara et al, 1994). 3.3.4.1.2 Gene Mutation The ability of chlorothalonil to induce mutation at the thymidine kinase (TK) locus of mouse lymphoma L5178Y cells was examined by selection with trifluorothymidine. Three experiments involving 4 hour incubations were carried out in the absence of a metabolic activation system. Chlorothalonil proved to be highly cytotoxic, with concentrations of 0.24 g ml-1 and above being lethal. In the first experiment, the top dose (0.32 g ml-1) could not be analysed due to lethality, and there was no mutagenic response at the next lowest dose (0.16 g ml-1) where the growth relative to the negative control (RTG) was 58%. In the remaining two tests, number of mutant cells were increased by 2.6 times at each of the top doses which were 0.12 g ml-1 (42% RTG) and 0.2 g ml-1 (34% RTG). Thus there was a sharp response threshold for the positive results, which were only seen at levels of significant cytotoxicity. The cells responded appropriately to the positive control (McGregor et al, 1988). 3.3.4.1.3 Sister Chromatid Exchange (SCE) The ability of chlorothalonil to induce SCEs has been investigated in Chinese hamster ovary cells. The treatment times were 2 hours and 25 hours with and without Aroclor-induced rat liver S9 respectively. The doses were chosen such that the top concentration reduced growth by approximately 50%. Without S9 (range 0.4 to 0.5 g ml-1, 28 hour harvest time.) the result was negative. Two tests were performed with S9. The first (range 0.25 to 2.5 g ml-1, 28 hour harvest time) showed a significant (25%) increase in SCEs over controls at the top dose only. The second gave 26% and 33% increases over controls at the mid and top doses (range 1.5 to 2.5 g ml-1) in a dose related manner, although within this experiment the fixation times were extended from 25.8 hours (negative control and low dose) to 33.3 hours (mid and high dose). The extended fixation time seems to have been due to cell cycle delay, and certainly the positive results were only seen at toxic doses. As with other tests there was a steep response curve. Expected results were gained with the positive controls (Galloway et al, 1987). 3.3.4.1.4 Single Cell Gel Electrophoresis (Comet) Assay This assay, which has been developed to measure DNA damage either in vitro or in vivo, is a respected test although it has not yet been internationally validated. It has been used to assess the genotoxic potential of chlorothalonil in vitro in peripheral blood human lymphocytes and Chinese hamster ovary K1 cells. In the experiment with human lymphocytes, the cells were incubated for one hour with 10 to 500 M chlorothalonil, followed by assessment of DNA damage and general cytotoxicity. There were appropriate negative and positive controls. Immediately after the 1 hour incubation, the cell viability was 90%, but 24 hours later, after incubation in fresh medium, this had dropped to 60% with 10 M and 30% with 500 M. DNA damage was detectable at the lowest dose upwards when assessed immediately after the 1 hour incubation, although an increase in the proportion of damaged cells was only detectable at and above 50 M. After 24

15

hours, the degree of DNA damage had increased dramatically, with 90% of cells damaged or highly damaged at the lowest dose and 100% death at and above 50 M. Further testing revealed the existence of a sharp threshold; lowering of the chlorothalonil concentration from 10 to 5 M led to the disappearance of both DNA damage and cytotoxicity. Thus the DNA damage had occurred only as a step towards cell death, and the chlorothalonil was not specifically genotoxic (Lebailly et al, 1997). The Chinese hamster ovary cells were likewise treated for 1 hour with chlorothalonil, but in this case viability was measured either immediately by dye exclusion (membrane integrity), or 2 days later by a test for oxidative activity. The concentrations chosen were 0.1 to 1.0 M, with the top dose causing approximately 25% cytotoxicity as measured by oxidative activity (i.e. the activity was 75% of the negative control), although cell viability as measured by dye exclusion was 100% when measured immediately after incubation. This is consistent with the earlier lymphocyte work where chlorothalonil had a delayed effect on frank cytotoxicity. There was some evidence of DNA damage at and above 0.2 M, and although there was a positive dose response, increases were small. This study indicates that chlorothalonil may be specifically genotoxic to Chinese hamster ovary K1 cells in vitro, but that it is difficult to exclude oxidative damage even at weakly cytotoxic concentrations (Vigreux et al, 1998). 3.3.4.1.5 DNA Adduct Formation The ability of Daconil, a formulation containing 75% chlorothalonil, to induce the formation of DNA adducts was investigated in the presence of Aroclor-induced rat liver S9 as an external metabolising system. The metabolites were extracted (though not identified) and then incubated with calf thymus DNA for 3.5 hours prior to purification and digestion of the DNA. The adducts were enriched by either nuclease P1 digestion or butanol extraction, post-labelled with 32phosphate and then chromatographed. The use of both enrichment methods on separate samples increased the likelihood of identifying adducts. Two independent experiments were carried out. Daconil failed to induce an increase in DNA adducts compared to negative controls; the positive control, however, caused a very large increase. This study shows that the formulation Daconil is not active in DNA adduct formation (Shah et al, 1997). 3.3.4.2 In Vivo Studies 3.3.4.2.1 Chromosome Aberration Studies As discussed in the introduction, earlier work in Chinese hamsters showed a small, but nondose-related increase in aberrations after treatment with chlorothalonil, a result which was considered to be negative. Due to concerns elsewhere, the authors carried out further work on repeated dosing of rodents with chlorothalonil; although the full studies have not been published, summary data are available. The dosing regime for Chinese hamsters and rats consisted of five consecutive oral doses and multiple harvest times 6 and 24 hours after the last dose. The Chinese hamsters (9 to 10 per group) received 0, 187.5, 375 or 750 mg kg-1 d-1 and the rats (5 per group) received 0, 500, 1000 or 2000 mg kg-1 d-1. This compared to the original study in which Chinese hamsters received 0, 50, 125 or 250 mg kg-1 d-1 for 5 days. Between 250 and 500 bone marrow cells were evaluated for each dose and harvest time. The authors stated that the results in both species were clearly negative, and the summary tables giving the percentage of cells with breaks confirmed this (Mizens et al, 1998).

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3.3.4.2.2 Oxidative DNA Damage Oxidative damage to DNA from rat liver can be investigated by comparing the level of 8hydroxy-2-deoxyguanosine (8-OH-2DG) to that of 2-deoxyguanosine (2DG) in the isolated DNA. Lodovici has studied this effect in Wistar rats exposed to a range of pesticides, as mixtures or by themselves; only the experiment with chlorothalonil alone is discussed here. Groups of 5 or 6 rats were dosed orally by gavage on 10 consecutive days with 0.1, 0.13, 0.5 or 1.0 mg kg-1 d-1 chlorothalonil in corn oil. There was also a group of 17 untreated control rats. The rats were then killed and the DNA isolated from the excised liver. This DNA was digested and analysed by high performance liquid chromatography, which separated 8-OH2DG from 2DG, the former being a measure of oxidative damage, possibly due to the production of free radicals. Chlorothalonil induced a dose-dependent increase in DNA damage at 0.13 to 1.0 mg kg-1 d-1, with the mean at the highest dose being about 4.5 molecules of 8-OH-2DG per 105 2DG nucleotides compared to about 1.5 in negative controls. This is consistent with the findings seen in vitro during the Comet assay, above, and its mode of action as a fungicide in which it reacts with cellular sulphydryl groups [as discussed in ACP (1994)]. However, the study did not report whether there was general cytotoxicity at the doses causing DNA damage. Since no liver pathology has been observed in lifetime studies at up to 175 mg kg-1 d-1 in rats, as assessed in ACP (1994), the significance of these findings in the intact animal is unclear (Lodovici et al, 1997). 3.3.4.3 Summary Positive results have been obtained with chlorothalonil in internationally recognised mammalian cell in vitro genotoxicity tests. However, these results have been found at cytotoxic doses with a profile indicative of DNA damage being secondary to cellular toxicity. They should also be compared against a number of negative in vitro and in vivo studies already assessed in the 1994 review of the agricultural and horticultural uses of chlorothalonil [ACP (1994)]. Although oxidative damage to DNA has also been recently demonstrated in vivo, this is again related to the cellular toxicity of chlorothalonil, and there is no evidence that this causes any long-term effects in the intact animal. Further chromosome aberration assays using Chinese hamsters and rats have confirmed the absence of genotoxic potential in vivo. 3.3.5 CHRONIC TOXICITY/CARCINOGENICITY Chlorothalonil produced an increased incidence of hypertrophy / hyperplasia and adenomas / carcinomas of the proximal renal tubule epithelium in rats and mice. There was also evidence of renal tubular hypertrophy in 2 y dog studies. The critical NOELs for effects on the kidney were 2 mg kg-1 d-1 for renal tubule hyperplasia and 4 mg kg-1 d-1 for renal tumours from a 2 y rat study (ACP, 1994). Chlorothalonil also caused an increased incidence of hyperplasia and hyperkeratosis of the forestomach epithelium and an increased incidence of papillomas and carcinomas in the forestomach of rats and mice. The critical NOELs for effects in the forestomach were 1.86 mg kg-1 d-1 for hyperplasia from a 2 y mouse study and 4 mg kg-1 d-1 for forestomach tumours from a 2 y rat study (ACP, 1994). No studies on carcinogenicity have subsequently been published that would alter the position that the committee reached. 17

3.3.6 REPRODUCTIVE TOXICITY Chlorothalonil was not teratogenic in the rat or rabbit. There was no effect on reproductive performance in rat multigeneration studies (ACP, 1994). 3.3.7 HUMAN DATA The results of an annual screening programme at a plant manufacturing chlorothalonil revealed that of the 191 employees participating in the screening, 5 had a rash (of these 3 had no skin abnormalities), 1 had a history of psoriasis and 1 had a rash localised to the external genitalia. Another survey was conducted at the same plant in 1978 and 1979. At the end of 1978 engineering controls were introduced in the packaging system. In 1978, there were 157 participants (67 % of the workforce); 62 cases of skin abnormalities were reported including 19 cases of contact dermatitis in 103 workers handling chlorothalonil, and 2 cases of contact dermatitis in workers not exposed to chlorothalonil. In 1979, there were 148 participants (68 % of the workforce); of the 123 workers exposed to chlorothalonil, 26 had skin abnormalities. There were no reported cases of contact dermatitis (ACP, 1994). Analysis of medical records of 43 workers from a Japanese chlorothalonil plant between 1971 and 1978 revealed 11 cases of reported dermatitis (ACP, 1994). Health screening reports from an American chlorothalonil plant were submitted for the years 1979 - 81, 1980 - 82 and 1981 - 1985. Between 1979 - 81 there were few changes in clinical chemistry, haematology or urinalysis findings. There were no significant changes in the reports for 1980 - 1982 or 1981 - 1985. There was one report of dermatitis in 1980 (ACP, 1994). An updated search of the published literature has identified the following studies: During the course of their work constructing window frames, 20 Norwegian workers used a wood preservative containing 0.5% chlorothalonil in white spirit, a polyvinylacetate glue resin and a chromium nitrate hardener. Fourteen of these workers reported symptoms of pruritis, erythema and oedema of the eyelids, with some experiencing a spread of the dermatitis to the face, neck and forearms. The symptoms were clearly related to work at the factory. All 14 were patch tested with a standard series, glue resin (5% in water), hardener (1% in acetone) and chlorothalonil (0.01% in acetone). Seven were found to give positive test reactions to chlorothalonil ranging from a few papules to marked papular erythema, 6 gave negative responses, and one reaction was reported as toxic. None of the subjects reported any reaction to the glue or hardener. Only one of 14 control subjects tested with 0.01% chlorothalonil showed a positive response. Overall, this study provides reasonable evidence that chlorothalonil may have the potential to cause allergic contact dermatitis in humans (Johnsson et al, 1983). Over a period of 2 years, 2 carnation nursery workers and a manager who had contact with several flowering plants were examined for contact dermatitis. All three had come into contact with chlorothalonil, and were patch tested with a standard series, plant extracts and various pesticides. The first subject was a man who had worked in a carnation nursery without experiencing skin problems for 10 years, but then developed a widespread dermatitis, and had three attacks of generalised pruritis and urticaria. The only positive reaction on patch 18

testing was to 1% chlorothalonil which, as results for controls described below indicate, is too high a concentration to allow differentiation between irritant and sensitisation reactions. The second case concerned a woman who developed dermatitis of the hands, arms and neck after a period of several years of working in a carnation nursery. Positive patch test reactions were obtained with 0.01% chlorothalonil and carnation leaves sprayed with chlorothalonil. The third subject was the male manager of a flower nursery who had periods of mild hand dermatitis, attributed to his atopic constitution, every winter for 23 years. When the dermatitis became more severe and longer lasting, he was patch tested and the only positive reaction obtained was to 0.01% chlorothalonil. For comparison, 17 control subjects were patch tested with 1, 0.1 and 0.01% chlorothalonil. The 2 higher concentrations produced clear delayed irritant reactions; at 0.01% there were no positive reactions but a marginal irritant effect was obtained in 4 cases. This type of reaction did not apparently interfere with the detection of the true positive reactions seen in 2 of the test subjects. Overall, these findings provide further evidence of the skin sensitising potential of chlorothalonil in humans (Bruynzeel and van Ketel, 1986). A questionaire-based health survey of 59 male and 48 female flower growers was carried out shortly after the spring chrysanthemum harvest season, with a total of 28 subjects complaining of work-related skin symptoms. Patch tests were conducted with a series of pesticides used in flower-growing and with chrysanthemum extracts; chlorothalonil was administered as the formulation Daconil at a final concentration of 0.015%. None of the materials applied caused irritant or allergic reactions among a control group of healthy, nonagricultural subjects on preliminary patch testing. The incidences of positive reactions on patch testing in relation to chlorothalonil were 3/59 males and 6/48 females (Ueda et al, 1994). Nineteen men and 9 women employed in a chlorothalonil manufacturing plant were investigated for occupational respiratory and skin disease. Exposure to chlorothalonil was for approximately 8 hours daily, six days a week at an average airborne concentration of 0.72 mg m-3. Two cases of possible skin sensitisation were identified by consideration of skin symptoms and the results of patch tests with 0.05% chlorothalonil, while none of the control group of 12 male and 6 female office and repair workers reacted to chlorothalonil. It was also noted that the chlorothalonil workers suffered increased respiratory symptoms compared with those reported by the controls (Huang et al, 1995). A group of 39 banana plantation workers was selected from an occupational dermatology clinic in Panama for the presence of a chronic pigmented dermatitis. Findings of positive patch test reactions to 0.001% chlorothalonil in 34 of these patients, as compared to no reactions in the control group of 41 patients attending the clinic with other types of dermatitis, indicate that the pigmented dermatitis was possibly related to exposure to chlorothalonil (Penagos et al, 1996). In another study, chlorothalonil was one of a group of pesticides investigated as a suspected cause of dermatitis in California - during 1982 - 1989, 10 cases associated with chlorothalonil use were reported in an occupationally exposed population. When groups of 39 nursery workers occupationally exposed to a range of pesticides and 21 unexposed control subjects were patch tested with 0.001% chlorothalonil, there were no positive responses; 4/11 controls reacted to 0.01%, indicating an irritant reaction. Since it is unclear whether or not those tested actually had skin disease, this report provides very limited useful information in relation to chlorothalonil (OMalley et al, 1995). 19

There are also a number of case reports available. One describes a male cabinetmaker who developed a dermatitis on his hands, which gradually spread to his arms and face, 9 months after starting work involving the use of preservatives containing 0.5% chlorothalonil. Patch tests resulted in positive reactions to chlorothalonil at 0.01% and at 1%, the latter covering an area of some 15 by 30 cm. Adverse skin reactions were noted at 1% (2/2) and at 0.1% chlorothalonil (2/10) in controls, but 0.01% gave no reactions in 10 controls. The dermatitis disappeared after 3 weeks sick leave but reappeared on return to work (Bach and Pedersen, 1980). Another case concerns a male painter, with no history of skin disease, who developed repeated episodes of dermatitis on his face and arms after using a paint (Bett) containing about 0.1% chlorothalonil. Tiny drops of paint got on his skin while working. A patch test with a standard series was negative, while patch tests with the paint formulation produced a strong reaction, and there were also positive reactions to chlorothalonil down to 0.0001% (Meding, 1986). Acute dermatitis consisting of widespread oedema and suspected urticaria developed in a man who slept in a cottage where a month earlier he had painted the window frames with a paint containing 1% chlorothalonil. When the man vacated the cottage the dermatitis disappeared, but it subsequently reappeared on his return, even as long as nine months later. Patch testing was carried out with a series of products owned by the patient and with the nine ingredients of the suspect paint. For his own products the patient tested positive to the suspect paint at concentrations down to 0.312 %, while 6 other paints and a glue gave negative results. When patch tested with the nine ingredients of the suspect paint, the patient was positive only to chlorothalonil, at concentrations of 0.0001% and above. Control tests using 0.0001% chlorothalonil carried out in 10 dermatitis patients were negative (Lidn, 1990). A woman employed in a redwood plant nursery presented a recurrent pruritic eruption involving facial erythema and marked edema, which occurred within 15 to 30 minutes of her arrival at work. She had indirect contact with a number of pesticides, including several chlorothalonil formulations, through contact with the treated foliage of seedlings. Patch testing to a standard series and to a pesticide series that did not include chlorothalonil revealed a positive response only to nickel. Immediate tests were then carried out by applying samples of Douglas fir and redwood seedlings treated with a chlorothalonil formulation to normal skin for five minutes, resulting in a significant wheal and flare reaction. Plants that had not been treated with chlorothalonil failed to elicit a response when applied to both intact and pricked skin. In a subsequent test, application of a 0.01% dilution of the chlorothalonil formulation to intact forearm skin resulted in the appearance of a large wheal and flare, followed by marked facial flushing, eyelid oedema, and difficulty in swallowing and breathing. Thus this study suggests that chlorothalonil can cause an immediate-type hypersensitivity response, consisting of contact urticaria and anaphylaxis, in addition to the delayed-type skin sensitisation response reported elsewhere (Dannaker et al, 1993). Finally, a case of photoallergic dermatitis has been attributed to the use of chlorothalonil. A man was exposed through gardening to a variety of pesticides, including a chlorothalonil formulation called Daconil, and developed an eruption of light-exposed skin occurring repeatedly from spring to summer. Patch and photopatch tests were carried out with a standard series, pesticides including Daconil and other potential photosensitisers. Positive reactions on both patch and photopatch testing were found only to Daconil (0.002%), with a 20

stronger reaction being observed to the photopatch test, and the patient remained sensitive to sunlight even after the Daconil had not been used for over 12 months. However, 5 of the 20 control subjects gave a positive patch test to 0.002% Daconil, and 4 a positive photopatch test, so that no firm conclusions can be drawn about the nature and cause of the skin condition in the gardener (Matsushita et al, 1996). Following a search of the relevant databases, one case of allergic contact dermatitis to chlorothalonil in a packer was found in the OPRA (Occupational Physicians Reporting Activity, May 1994 to 1998) database and another case of contact dermatitis due to chlorothalonil in a line operator was found in the EPIDERM (Occupational Skin Surveillance Survey, 1993 to 1998) database. In addition to the above information, there have been no incidents related to chlorothalonil reported to HSE under RIDDOR (the Reporting of Injuries, Diseases and Dangerous Occurrences Regulations) in the last 14 years. Similarly, HSEs SWORD (Surveillance of Workers for Respiratory Disease) database had no records of any incidents related to the use of chlorothalonil. No incidents associated with chlorothalonil-containing antifouling products were reported to the Pesticides Incidents Appraisal Panel (PIAP) between 1989 and 1999. The Belfast and Edinburgh Centres of the National Poisons Information Service have reported that they received no enquiries concerning chlorothalonil in the periods from 1996 onwards and 1998 onwards, respectively. The Cardiff Centre reported very brief details of 4 incidents related to antifouling products in the period 1997-1999 but was unable to relate these to specific products or active substances. 3.3.7.1 Summary Studies on groups of workers and individuals occupationally exposed to chlorothalonil indicate that it has the potential to cause allergic dermatitis in humans, and there is also some, more limited evidence that it may cause contact urticaria.

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OPERATOR AND CONSUMER EXPOSURE AND RISK ASSESSMENTS

4.1 NO ADVERSE EFFECT LEVELS FOR USE IN RISK ASSESSMENTS

4.1.1 PROFESSIONAL USE

A review of the agricultural uses of chlorothalonil was considered by the ACP in January 1994 (ACP, 1994) and a NOEL of 3 mg kg-1 d-1 was agreed, derived from a 13 week study in Sprague Dawley rats and based on hyperplasia and hyperkeratosis of the forestomach at the next highest dose. This NOEL will be used in the risk assessments for systemic toxicity to professional operators (including chandlers) together with a default value of 10 % for dermal penetration. The detailed discussion on the areas of toxicological concern are reproduced in italics from ACP (1994). RISK ASSESSMENT The two issues of concern with chlorothalonil are the occurrence of stomach tumours of the non-glandular epithelium and tumours of the renal proximal convoluted tubule. For genotoxic carcinogens, where any human exposure would be a cause for concern, it is generally accepted that no safety factor, however large, can be considered to provide adequate assurance of safety. The mutagenicity studies reviewed in this document provide clear evidence that chlorothalonil is not genotoxic. In order to provide reassurance for consumer and operator safety it is necessary to establish the underlying mechanism when dealing with non-genotoxic carcinogens. Areas of Toxicological Concern Stomach tumours The stomach tumours were associated with a high incidence of hyperkeratosis/hyperplasia of the non-glandular epithelium. Although humans do not possess a forestomach the lesions observed cannot be ignored since the lower oesophagous in man has the same embryonic origin as the forestomach. However, the incidence of stomach tumours can be explained in terms of the irritant nature of chlorothalonil. Chlorothalonil is a severe irritant and it is probable that the hyperplasia and hyperkeratosis seen in the rat forestomach is a response to the irritant insult by chlorothalonil. One of the consequences of the induced hyperplasia in the forestomach is that a larger population of cells will be at risk from spontaneous mutational events. There is a substantial body of evidence that chlorothalonil is not genotoxic in addition there is clear evidence of a threshold below which the observed stomach tumours are not induced. It is possible to establish a no-effect level based on the induction of hyperplasia/hyperkeratosis of 1.8 mg-1 kg1 day-1 from a 2 year mouse feeding study.

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Renal tumours The renal toxicity induced by chlorothalonil is quite typical of several halogenated alkenes such as hexachloro-1,3-butadiene and fluorinated ethylenes. In all cases the molecule is coonjugated with glutathione then further metabolised in the kidney to cysteine conjugates by glutamyl transpeptidase then further metabolised to thiol metabolites by cysteine conjugate lyase. studies have located the pars recta as the site of cysteine conjugate lyase. localisation in the proximal tubule. Studies have shown that chlorothalonil is excreted via the kidney as thiol metabolites. Treatment of rats with GGT inhibitor AT-125 lead to an accumulation of radiolabel in the kidney. It has been shown that in vitro chlorothalonil is only absorbed from the duodenum as glutathione conjugates or further metabolites. In addition it has been shown that in vivo the level of radiolabel found in urine and kidney of germ free rats dosed with 14C chlorothalonil was lower than in non-germ free rats suggesting that the gut flora plays a role in the absorption of chlorothalonil. In vitro studies have shown that the di-thiol metabolite of chlorothalonil is toxic to kidney mitochondria. A possible mechanism for the formation of kidney tumours in the rat is the formation of the di-thiol metabolite of chlorothalonil in the kidney following the action of lyase on the cysteinyl conjugate of chlorothalonil. The di-thiol is toxic to renal tubule epithelial mitochondria leading to cell death and a consequential hyperplasia of the renal tubule epithelial cells. As a result of the increased number of target cells exposed to a constant background spontaneous mutation rate an increased incidence of adenomas and carcinomas of the renal tubule epithelium. The company have established that it is the di and tri-thiol metabolites that are the causative agents producing renal hyperplasia. Both metabolites inhibit the transfer of reducing equivalents from succinate to coenzyme Q in the electron transport system. this is likely to result in reduced levels of ATP and a consequent impairment of the Na/K pump. When all the chronic studies are considered together the results suggest that the kidney tumours might be strain specific. It would appear that Fischer 344 and Osborne Mendel rats are more sensitive than Sprague Dawley rats. In particular there is no evidence of kidney tumours in the Sprague Dawley rat even at dose levels of 1500 mg-1 kg-1. It has already been shown that chlorothalonil is only absorbed as the glutathione conjugate. A possible explanation for the apparent strain specificity could be the different gut flora between strains. This could also explain why primates absorb less chlorothalonil than rats and points towards a possible species specificity for the renal toxicity of chlorothalonil. In 1992 the JMPR confirmed the ADI of 0.03 mg-1 kg-1 for chlorothalonil. The report will clarify their rationale for this ADI: that the current information suggests that the rat data can be ignored in favour of the dog in setting an ADI for humans. The 1992 JMPR meeting was also made aware of further data being generated by the registrant, including metabolism studies in dogs which will help to strengthen the conclusion regarding species specificity of the renal toxicity of chlorothalonil.

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Consideration of an Acceptable Daily Intake It is possible to establish a no effect level based on the induction of hyperplasia/hyperkeratosis in the mouse forestomach of 1.8 mg-1 kg-1 d-1 in a 2 year mouse feeding study. The ACP recommended a safety factor of 100 for these effects, on this basis an Acceptable Daily Intake for consumers of 0-0.018 mg-1 kg-1 d-1 can be established. Consideration of an No Adverse Effect Level In considering the risk to operators it is important to consider likely levels of thiol metabolite formation following dermal exposure. In the rat, following oral dosing at 50 mg-1 kg-1, 1.6 % of the applied dose was excreted in urine as thiol metabolites. Following dermal dosing the rat excreted 0.07 % of the applied dose as thiol metabolites. In Rhesus monkeys following oral dosing the tri-thiol metabolite was found in urine at up to 0.01 % of the applied dose. Following dermal application less than 0.008% of the applied dose was excreted as thiols. In terms of dermal absorption the primate can be considered as more representative of the human situation than the rodent data. It can therefore be assumed that the levels of thiol metabolites produced in humans exposed dermally will be at least 200 times less than in rats dosed orally. Although there are indications that, in this instance, the rat is an inappropriate model the no effect levels from rat studies are being used until the registrant has made a case to substantiate this conclusion. In a 13 week study a no effect level of 3 mg-1 kg-1 was established based on kidney weight changes at 10 mg-1 kg-1 d-1. Hyperplasia was also noted at 40 mg-1 kg-1 d-1. In view of the fact that the incidence of kidney tumors appears to be strain specific and the fact that there is no evidence that chlorothalonil is genotoxic it is proposed that the no effect level from the 13 week rat study should be used in setting an AOEL. Since thiol levels in primates dosed dermally are at least 200 times lower than in rats dosed orally it is suggested that the AOEL should be 0.03 mg-1 kg-1bw d-1 and using a dermal absorption figure of 0.5 %. 4.1.2 AMATEUR USE It is considered inappropriate to estimate the risks to amateurs by comparison of exposure with either a LD50 value or NOAEL from an acute oral toxicity study, since these studies do not require a full histopathological assessment. No oral studies of less than 90 d duration are available and therefore amateur exposure has been compared with the NOAEL of 3 mg-1 kg-1 d-1 derived from a 13 week study in the rat and already used in the risk assessment for professional users. It was agreed that the risk assessment for inhalation exposure should use the acute LC50 value. Two studies on the acute oral inhalation toxicity of chlorothalonil are available and LC50 values of 0.31 mg l-1 (1 h) and 0.11 mg l-1 (4 h) have been reported. The latter (equivalent to 110 mg m-3) is considered most appropriate for comparison with worker exposure. Calculations for inhalation risk, for both professionals and amateur workers, were provided. 24

A further consideration is that of skin sensitisation. Although no conclusions could be made from the animal studies submitted, human data indicate a potential for skin sensitisation.

4.2 OPERATOR AND CONSUMER EXPOSURE

4.2.1 INFORMATION RELATING TO USER EXPOSURE Risk assessment patterns of use data are based on surveys and anecdote. They are as accurate as is possible, but should not be regarded as definitive. 4.2.1.1 Industry Data Company No. 2 have submitted the following study: Potential exposure of workers to chlorothalonil when handling and applying paint containing chlorothalonil (Unpublished, 1995). This study did not involve the actual spraying of ships, however, it did involve a similar task and was considered adequate for risk assessment purposes. 4.2.1.2 HSE Data Since 1994, the Health and Safety Executive has gathered information on human exposure to antifouling products in the professional and amateurs sectors, to inform its role of assessing exposure and risk to operators and others. The information takes two forms: the pattern of work - the frequency and duration of potential exposure, the areas coated per session, the amount of product used and seasonal factors; the exposure - the median and realistic worst case exposures in applying the products and identified tasks or jobs.

The surveys and studies informing HSE assessments are: 9 surveys applying copper-based antifoulant to ships (40 exposure data; HSE, 1994) 5 surveys applying tin-based antifoulant to ships (20 exposure data; HSE, 1996) 4 surveys applying various antifoulant to ships (10 exposure data; IOM 1996) pattern of use survey (2 commercial organisations, 4 service organisations for leisure craft; HSE 1994/5) 8 surveys applying copper-based antifoulant to leisure craft (9 exposure data; HSE, 1997/9)

All HSE surveys took place in the north of England or Scotland. Some information on patterns of use was derived through exposure surveys. Each exposure data point comprised the potential dermal exposure (the amount of antifoulant product depositing on the outer surface of the person), the exposure of hands inside protective or other gloves, exposure by inhalation, the tasks done and the amount of product used. HSE holds no information on the relative market importance of products or active substances. Three-quarters of the products found being used in the 1994/5 survey were free-association (conventional or contact leaching) (i.e. the active ingredient leaches from the antifoulant) and one-quarter self-polishing (ablative) (i.e. the active ingredient is bound in a copolymer antifoulant which hydrolyses slowly in sea water). HSE has no reliable data relating to

25

exposures in the military sector where data relate only to airborne concentrations of copper. There are no data for other immersed structures (e.g. oil-rigs, jetties, fish-farm installations), nor on exposure in stripping expired antifoulings. 4.2.1.3 Environment Agency Data The Environment Agency (1998) commissioned a report on environmental problems from antifouling agents that contained information on patterns of use. 4.2.1.4 Expression of Exposure Data All HSE data are quoted in terms of the antifoulant product being applied and are timeweighted. Data are therefore normalised and in the forms mg product h-1 for dermal exposure and mg product m-3 for inhalation exposure, respectively [HSE (2000)]. The sampling methods for potential dermal exposure using patches have been validated for spraying activities (Unpublished, 1996; Unpublished, 1998). However, they have not been validated for painting or paint handling. It is inappropriate to express exposure simply as a single value. In fact, there are exposure distributions. However, data are sparse due to the difficulty and high cost of their acquisition, and the exact nature of the distribution cannot be proven. Consequently, complex statistical treatments are considered inappropriate. HSE statisticians gave their opinion that in cases where data are sparse, it is valid to consider in detail only the non-zero results. Results with an effective value of zero can be taken into account in the frequency or chance that exposure will occur. The median value in a distribution, moderated by this frequency, represents a central tendency value. A realistic worst case is represented by the 95th percentile data point of nonzero values, or if the distribution data are sparse, by the highest values found. Where the highest data point is a clear outlier (e.g. many times higher than the next highest point), a decision can be taken to note that datum but disregard it for the purposes of risk assessment. 4.2.1.5 Mitigation of Exposure to Antifoulants The work clothing, whether or nor constituting formal personal protective equipment, does have protective properties. Impermeable clothing (e.g. Tyvek suit) will stop liquids reaching the skin, but easily becomes contaminated inside and is difficult to clean properly. Furthermore, deposits may concentrate on the equipment surface and are available for dislodging. Cloth-based equipment will tend to retain liquids and dusts to some extent; there will also be run-off. Penetration of clothing occurs through liquids soaking through seams, zips and elasticated parts, through being rubbed through by frequent contact and by being drawn through openings at neck, wrist and ankle by the bellows effect when the operator moves inside the clothing. HSE data indicate a median 4% penetration of the outer layer of work clothing, generally a coverall. Having more than one layer of work clothing will provide better protection. Protective gloves are generally effective in protecting the hands from antifoulant. However, hand contamination is inevitable, and the exposure route may be through putting on and taking off used gloves. One measure to mitigate continuing exposure through protective

26

gloves being contaminated inside is for regular replacement of protective gloves, for example following each antifouling job. Respiratory protective equipment (RPE) needs to achieve two objectives: to mitigate exposure by inhalation and to protect the skin of the face, head and neck. In general, a high standard of respiratory protective equipment with a workplace protection factor of at least 50 is needed for sprayers and possibly other RPE for some ancillary workers,. The person at greatest risk is the sprayer, and for spraying, such protective equipment should be mandatory. If the pot-men were to work in the vicinity of the spray plume, then COSHH is likely to indicate that they should wear appropriate protective equipment. Appropriate equipment would probably be of a standard equivalent to an FFP3 disposable filtering face piece respirator or better. However, the need for respiratory protection for workers other than sprayers is considered to be a matter for the COSHH workplace risk assessment. It is considered unlikely that amateur users would be able adequately to select and use RPE. 4.2.1.6 Factors Affecting Exposure Estimates for the Use of Antifouling Products When the partial reviews of the physical chemistry and mammalian toxicology of the booster biocides TCMTB, zineb and zinc pyrithione were presented to the ACP in 1999, concern was expressed over the apparently low values of the Toxicity Exposure Ratios (TERs) derived for a number of the compounds. It was agreed that the assessment of risk to users and bystanders should be revised to take into account all factors affecting exposure and increase the clarity and transparency of the risk assessments. The TERs of particular concern related to the high end exposures (95th percentile or higher) that were calculated from the operator exposure model. For antifouling applications, the high end predictor is based on the 95th percentile for sprayers and the worst case results for pot-men. Dermal Exposure HSE believes the antifouling model to be an accurate predictor of the amount of product that deposits on the outer surfaces of a workers clothing - as a worst case this amounts to in excess of 120 g of product in a spray session. What is available for uptake following transfer to the skin is another matter. The estimation of systemic dose, resulting from conditions that can lead to 120 g of paint on the outside of clothing is open to interpretation. It requires a degree of professional judgment to reach a realistic conclusion. (The dermal absorption value used in the risk assessment is also critical to the resulting estimate of systemic dose). Extent of Penetration through Clothing HSE data indicate that 4 % is a realistic figure to adopt for the penetration of antifouling products through a single layer of typical protective workwear. Information from a number of sources suggests that the performance of protective clothing may be related to the level of the challenge. Laboratory experiments indicate that the higher the challenge, the lower the proportion of product that will penetrate to become available as a potential source for contact with the skin. This phenomenon has also been observed during field studies, such as those reported by the Institute of Occupational Medicine (1994) during the investigation of sheep dipping practices and field-effectiveness of PPE. Further layers of clothing will provide an extra barrier to skin contact.

27

Custom within the antifouling industry, brought about by a recognition that formulations may be unpleasant to work with and are difficult to remove from the skin, is for operators to protect themselves well, and often to wear two sets of coveralls. HSE is prepared to accept that it may be possible, at the higher levels of contamination, to consider a penetration factor through coveralls and clothing, and then onto the skin, at about 1 %. The value for penetration at 1 % is seen as the practical lower limit for modeling purposes as there will always be the potential for contact of product with exposed skin (e.g. around wrists, face and neck and through handling previously contaminated clothing). The ability of protective clothing to reduce potential exposures by at least two orders of magnitude has been demonstrated in the studies by the IOM and is supported by the experimental findings related to penetration compared to challenge which are built into the POEM model. Extrapolation from Clothing to Skin to Systemic Dose The modeling process is not very good at estimating how much of the product finds its way to the skin, and how much of the active substance in the product is eventually absorbed. The current HSE estimates are precautionary and based on all of the product that is predicted to penetrate the layers of clothing getting to the skin, and a quantity of the active substance within the product immediately penetrating through to become a systemic dose. (The amount of uptake via the skin may be established through dermal absorption studies, or more often by using a default value such as 10%). However, such uptake is considered unlikely to happen in reality for a number of reasons: w Only a proportion of what lands on the skin is available to be absorbed when product deposition occurs in the form of spots or blobs - active substance will be contained within the matrix of the dried-on product; The model does not take into account the dynamics of deposition or absorption, nor the kinetics of metabolism; The model does not take into account actions to remove residues after work, particularly from the hands.

w w

Patterns of Exposure A further factor relates to interpretation of the findings on exposure. Account needs to be taken of the pattern of work for a painter and the pattern of use of any particular product. Professionals spend much of their time carrying out preparatory work during vessel refitting many other jobs take place while a vessel is in dry dock and it may be there for a number of weeks. Consequently, exposure to antifouling paints is irregular with long intervals between exposure. Exposure to one particular active substance, other than copper compounds, is even less frequent. The most realistic worst case exposure scenario is that a painter may be exposed for no more than two or three days a month, but not every month, and then not to the same active substance. High end exposures do not occur every time and could be considered as acute.

28

Hand Exposure Estimates of hand contamination play an important part in development of the exposure assessment for antifoulants. This is particularly true for the pot-men who come into contact with large amounts of product while replenishing reservoirs for sprayers. There is always the opportunity for spillage. Where pot-men have worn suitable and adequate new gloves, it appears the levels of contamination to the hands have generally been low. Where pot-men have worn inappropriate gloves, old gloves, or no gloves at all, elevated exposures have been registered. These elevated exposures heavily influence the model. Appropriate changing of gloves suggests that hand exposure can be reduced. For the sprayer, hand exposures tend to be lower than for the pot-man and are not a main driver of the exposure estimate. Inhalation Exposure Sprayers will always need to wear respiratory protective equipment. For the pot-men, exposure to spray aerosol is intermittent and unusual. Results indicate that the normal range (18 of 19 results) is between 0.2 and 4.0 mg m-3 of product. One exceptional result (the highest recorded) has been recalculated at 24 mg m-3 (previously 42 mg m-3, but closer inspection of the proportion of active material in the paint has caused abatement of this particular result). However, it is considered to be unlikely that even a result of this magnitude would be a true reflection of the personal exposure of a pot-man to aerosol. The individual result showed no elevated potential dermal exposure. HSE has judged that, when considered in the context of the other samples within the data set, the specific result of 24 mg m-3 did not reflect exposure to inhalable aerosol: the result was more likely to have a risen through direct transfer of antifoulant, and possibly by direct contamination. 4.2.2 EXPOSURE ASSESSMENT - ANTIFOULANT SPRAYING Industry data are taken from the study: Potential exposure of workers to chlorothalonil when handling and applying paint containing chlorothalonil (Unpublished, 1995). Four professionals were each monitored using 3 different paint preparations, 3 times per preparation. They used commercial spray equipment to apply between 16 and 18.5 litres (5 US gallons, nominal) for between 22 and 81 minutes. Monitoring was by cotton whole-body dosimeters worn beneath work clothing, 2 head-patch samplers, inner and outer glove samplers and combined GF/A-sorbent tube samplers. The sampling covered mixing and loading, and application. Clearance monitoring was also undertaken. The equipment used was airless spray equipment operated at about 2000 psi (140 Bar). In addition to the work wear, the painters used 3M 8709 respirators and eye protection. The goggles became fouled and stocking masks to protect the skin were worn instead. Mixing and loading comprised stirring the product in the can, inserting the feed tube and priming the pump. Spraying included work from a ladder. All the surfaces sprayed were vertical in orientation. The following data are taken from guidance document HSE (2000). For roller and brush painting, data presented in Garrod et al (2000) has been assumed to apply to professionals.

29

4.2.2.1 Patterns of Use in Spray Operations - Professional Users The work involves dry-docking for vessels the size of tugs and above, or docking on a hard surface for small fishing vessels. During the time for removing and reapplying antifouling, general overhaul and refitting takes place. Consequently, applying the antifouling is a minor proportion of the time spent working on the vessel. Workers remove and apply surface coatings over the ship, (e.g. bilges, holds), using for example, two-pack epoxy preparations. The pattern-of-use survey indicated that up to 10 % of employees' time might be spent in working with antifoulings. Professionals work year-round. The vessel is cleaned with a high-pressure water-jet (for selfpolishing coatings) or with abrasive grit (for erodible coatings). Bare metal surfaces are prepared with coatings such as corrosion inhibitors. The antifouling is then applied using airless spray techniques at up to 100 bar. Sufficient sprayers are employed to ensure that one coat is applied in one work day. Rarely are more than two coats applied. Applying antifoulant requires 2 to 4 persons per spray position. The three identified tasks are: spraying - the sprayer; mixing and loading - the pot-man (who prepares the antifoulant and ensures its supply to the high pressure pump), ancillary - the rein or tender men, who attend to keeping paint lines free and may also manoeuvre the mobile access platform (cherry-picker).

Antifoulings are applied on several days a month, for no more than two consecutive days a week. There would normally be one coating session per day. In the HSE surveys the duration of daily work ranged from 40 to 360 minutes per coating session (median 184 minutes), for each of the three identified tasks. There are only estimated data for the quantity of antifoulant used per spray session. The quantity used ranged from 25 to over 800 litres of antifoulant (median 240 litres). The vessel surface areas coated ranged between 600 and 4000 m2 (median 1600 m2). Where safety data sheets are supplied with products to professional end-users, this supply is often through the ship owner, who does not necessarily transmit the data sheet. Contractors rely on a compendium of data sheets which may become outdated. Contractors were found to have a default set of equipment, risk assessments and personal protective equipment that they use for most situations. In all surveys, overalls and gloves were found to be available for use, though these items were not always suitable. Respiratory protective equipment was always found to be available. Sprayers usually took steps to protect any exposed skin from antifoulants. 4.2.2.2 Exposure Data for Spray Operations - Professional Users Detailed calculations are presented in Appendix 1, Tables 1.1 to 1.7. These are based on a 3 hour shift (as outlined in paragraph 4.2.1.6 extent of penetration through clothing) a value of 4 % penetration of work clothing and a value of 1% for penetration of a double layer of work clothing, sprayers, pot-men and other operators such as tenders. Exposure data are quoted as in-use concentration of antifouling product on the skin and inhaled. 30

Industry data has been recalculated in the format familiar for risk assessment of nonagricultural pesticide products and is presented in Appendix 1, Table 1.11. Data derived from antifoulant spraying (HSE data) is compared with industry data in Table 4.1. Table 4.1 Comparison of HSE and industry data Central Tendency Industry 2.1 mg 0.02 mg 0.10 mg Realistic Worst Case HSE 13 mg 1.76 mg 0.59 mg Industry 6.12 mg 0.09 mg 0.16 mg

Sample Type

HSE Inside PPE* 1.8 mg Inside gloves 0.44 mg Inhaled 0.05 mg * includes head exposure

HSE data for inside gloves represents the effect of wearing previously contaminated gloves. The industry data by definition used clean gloves for each exercise. While the industry data give a basis for estimating exposure, the results should be viewed with caution because the data are closely grouped (4 subjects, 9 replicates) and the glove use pattern is idiosyncratic. Consequently, no separate risk assessment will be conducted on the industry data. The HSE data shows wider exposure distributions, derived as they are from individual surveys. Table 4.1 suggests that HSEs default value of 4 % for single layer PPE is not unrealistic. 4.2.2.3 Patterns of Use in Spray Operations - Amateur Users There is no information that amateurs use spraying to apply antifouling, though this mode of application may be permitted in the conditions of approval. Clearly, spraying of leisure craft would be of shorter duration than the central tendency around 3 hours, (see Section 4.2.4.1) which is typical for professionals. In contrast with application by brush and roller (Section 4.2.5.2) there is often a clear need for respiratory and skin protection in spray application of antifoulants, and it is unlikely that amateurs would be able to select or use personal protective equipment adequately. 4.2.2.4 Exposure Data for Spray Operations - Amateur Users While there are no directly related exposure data, the data model relevant to professional sprayers should apply, assuming no RPE. Amateur sprayers are assumed to wear a coverall and therefore clothing penetration of 4 % has been used in calculations (unlike amateur use of brush and roller in Section 4.2.4.2). 4.2.3 SYSTEMIC EXPOSURES IN SPRAY OPERATIONS 4.2.3.1 Calculations The data sets and exposure calculations are set out in Appendix 1, Tables 1.1 - 1.7.

31

Assumptions: 5 % active substance in the product 10 % dermal penetration 4 % or 1 % clothing penetration The calculation for sprayers includes respiratory protection by air-fed RPE with a 50 fold factor for respiratory protection. This is summarised in Tables 4.2 - 4.7. Calculations are presented for clothing penetration at 4 % and 1 % for standard worker and additional PPE, respectively as explained in Section 4.2.1.6. The calculation for professional sprayers includes respiratory protection by air-fed RPE with a 50 fold protection factor. For amateur spraying, calculations are presented in Table 4.8, with 4 % clothing penetration as explained in Section 4.2.2.4. A summary of Tables 4.2 - 4.8 and resultant TER values is presented in Table 4.9. Table 4.2 Contact and Systemic Exposure to Chlorothalonil for Professional Sprayers: 4% clothing penetration Exposure item
Amount of product in contact with skin (mg d-1)
[3 hour job, 4% clothing penetration, 6170 mg h-1 central tendency, 44700 mg h-1 95th percentile, 60 (median) & 241 (worst case) mg h-1 of product in-glove]

Central Tendency 920

Worst Case 6080

Percentage of active substance in product (%) Amount of chlorothalonil in contact with skin (mg d-1) Dermal absorption value (%) Systemic exposure to chlorothalonil via dermal route (mg d-1) Intake of product by inhalation with RPE (mg d-1)
[3 hour job, 1.25 m3 h-1 inhaled volume, central tendency 6 mg m-3, worst case 64.6 mg m-3 of product]

5 46 10 4.6 0.45

5 304 10 30.4 4.84

Amount of chlorothalonil inhaled (mg d-1) Total systemic exposure for 60 kg operator (mg kg d ) wearing RPE
-1 -1

0.02 0.08

0.24 0.51

32

Table 4.3

[3 hour job, 4% clothing penetration, 2940 mg h-1 central tendency, 15000 mg h-1 95th percentile, 35 (median) & 1380 (worst case) mg h-1 of product in-glove]

Contact and Systemic Exposure to Chlorothalonil for Professional Pot-men : 4% clothing penetration Exposure item Central Worst Case Tendency Amount of product in contact with skin (mg d-1) 458 5940

Percentage of active substance in product (%) Amount of chlorothalonil in contact with skin (mg d-1) Dermal absorption value (%) Systemic exposure to chlorothalonil via dermal route (mg d-1) Intake of product by inhalation (mg d-1) *, without RPE.
[3 hour job, 1.25 m3 h-1 inhaled volume,, central tendency 0.6 mg m-3 , realistic worst case 3.84 mg m-3 of product]

5 22.9 10 4.6 2.25 0.11 0.04

5 297 10 304 14.4 0.72 0.51

Amount of chlorothalonil inhaled (mg d-1)

Total systemic exposure for 60 kg operator (mg kg-1 d-1)

* excludes top data point which is considered an outlier. This operator had significantly lower dermal exposure (by a factor of between 10 and 30) than each of the others in the study and his inhalation exposure was higher than that recorded for his associated sprayer - this would not be expected and cannot be explained. HSE concluded the sample has become contaminated and, consequently, the point discarded.

Table 4.4

Contact and Systemic Exposure to Chlorothalonil for Other Professional Operators : 4% clothing penetration Exposure item Central Tendency 211 Worst Case 956

[3 hour job, 4% clothing penetration, 885 mg h-1 central tendency, 3470 mg h-1 95th percentile, 35 (median) & 180 (worst case) mg h-1 of product in-glove

Amount of product in contact with skin (mg d-1)

Percentage of active substance in product (%) Amount of chlorothalonil in contact with skin (mg d-1) Dermal absorption value (%) Systemic exposure to chlorothalonil via dermal route (mg d-1) Intake of product by inhalation (mg d-1)
[3 hour job, 1.25 m h inhaled volume, central tendency 0.8 mg m-3 , realistic worst case 4.8 mg m-3 of product]
3 -1

5 10.55 10 1.06 3 0.15

-1 -1

5 47.8 10 4.78 18 0.9 0.0.1

Amount of chlorothalonil inhaled (mg d-1)

Total systemic exposure for 60 kg operator (mg kg d )

0.02

33

Table 4.5

[3 hour job, 1% clothing penetration, 6170 mg h-1 central tendency, 44700 mg h-1 95th percentile, 60 (median) & 241 (worst case) mg h-1 of product in-glove]

Contact and Systemic Exposure to Chlorothalonil for Professional Sprayers : 1% clothing penetration Exposure item Central Worst Case Tendency Amount of product in contact with skin (mg d-1) 365 2060

Percentage of active substance in product (%) Amount of chlorothalonil in contact with skin (mg d ) Dermal absorption value (%) Systemic exposure to chlorothalonil via dermal route (mg d-1) Intake of product by inhalation with RPE (mg d-1)
[3 hour job, 1.25 m h inhaled volume, central tendency 6 mg m-3 , worst case 64.6 mg m-3, of product]
3 -1

5
-1

5 103 10 10.3 4.84 0.24 0.18

18.25 10 1.83 0.45 0.02

Amount of chlorothalonil inhaled (mg d-1)

Total systemic exposure for 60 kg operator (mg kg d ) wearing RPE

-1

-1

0.03

Table 4.6

[3 hour job, 1% clothing penetration, 2940 mg h-1 central tendency, 15000 mg h-1 95th percentile, 35 (median) & 1380 (worst case) mg h-1 of product in-glove]

Contact and Systemic Exposure to Chlorothalonil for Professional Pot-men : 1% clothing penetration Exposure item Central Worst Case Tendency Amount of product in contact with skin (mg d-1) 193 4590

Percentage of active substance in product (%) Amount of chlorothalonil in contact with skin (mg d-1) Dermal absorption value (%) Systemic exposure to chlorothalonil via dermal route (mg d-1) Intake of product by inhalation (mg d-1) *, without RPE.
[3 hour job, 1.25 m3 h-1 inhaled volume, central tendency 0.6 mg m-3 , realistic worst case 3.84 mg m-3 of product]

5 9.65 10 0.97 2.25 0.11 0.02

5 230 10 23 14.4 0.72 0.4

Amount of chlorothalonil inhaled (mg d-1)

Total systemic exposure for 60 kg operator (mg kg-1 d-1)

* excludes top data point which is considered to be an outlier. This operator had significantly lower dermal exposure (by a factor of between 10 and 30) than each of the others in the study and his inhalation exposure was higher than that recorded for his associated sprayer - this would not be expected and cannot be explained. HSE concluded the sample has become contaminated and, consequently, the point discarded.

34

Table 4.7

Contact and Systemic Exposure to Chlorothalonil for Other Professional Operators : 1% clothing penetration Exposure item Central Tendency 306 Worst Case 2030

Amount of product in contact with skin (mg d-1)

[1 hour job, 4% clothing penetration, 6170 mg h-1 central tendency, 44700 mg h-1 95th percentile, 60 (median) & 241 (worst case) mg h-1 of product in-glove]

Percentage of active substance in product (%) Amount of chlorothalonil in contact with skin (mg d ) Dermal absorption value (%) Systemic exposure to chlorothalonil via dermal route (mg d-1) Intake of product by inhalation without RPE (mg d-1)
[1 hour job, 1.25 m h inhaled volume, central tendency 6 mg m-3, worst case 64.6 mg m-3 of product]
3 -1

5
-1

5 101.5 10 10.15 80.7

15.3 10 1.53 7.5

Amount of chlorothalonil inhaled (mg d-1) Total systemic exposure for 60 kg operator (mg kg d )
-1 -1

0.38 0.03

4.04 0.24

Table 4.8

Contact and Systemic Exposure to Chlorothalonil for Amateur Sprayers: 4% clothing penetration Exposure item Central Tendency 131 Worst Case 644

[3 hour job, 1% clothing penetration, 885 mg h-1 central tendency, 3470 mg h-1 95th percentile, 35 (median) & 180 (worst case) mg h-1 of product in-glove

Amount of product in contact with skin (mg d-1)

Percentage of active substance in product (%) Amount of chlorothalonil in contact with skin (mg d ) Dermal absorption value (%) Systemic exposure to chlorothalonil via dermal route (mg d ) Intake of product by inhalation (mg d )
[3 hour job, 1.25 m h inhaled volume, central tendency 0.8 mg m-3 realistic worst case 4.8 mg m-3 of product)
3 -1

5
-1

5 32.2 10 3.22 18 0.9 0.07

6.55 10
-1

0.66 3 0.15 0.01

-1

Amount of chlorothalonil inhaled (mg d-1)

Total systemic exposure for 60 kg operator (mg kg-1 d-1)

35

4.2.3.2 Assessment of Risk During Spraying Table 4.9 User Summary of Tables 4.2 - 4.8 and resultant TER values TER Worst Case (mg kg-1 d-1) TER

Central Tendency (mg kg-1 d-1) Professional users (NOAEL 3 mg kg-1 d-1) 4 % clothing penetration Sprayer 0.08 Pot-man 0.04 Ancillary Worker 0.02 1 % clothing penetration Sprayer 0.03 Pot-man 0.02 Ancillary Worker 0.01 Amateur users 4 % clothing penetration Amateur sprayer 0.03

38 75 150 100 150 300

0.51 0.51 0.1 0.18 0.400 0.07

6 6 30 17 8 43

100

0.24

13

Systemic exposure of professionals is acceptable at the central tendency when PPE is such that clothing penetration can be estimated at 1%. However it gives cause for concern at the worst case. The exposure estimate has assumed a default of 10 % skin penetration for chlorothalonil and a better estimate of the skin penetration of chlorothalonil from representative formulations would enable the risk assessment to be refined. It is therefore recommended that this be identified as a data requirement. The systemic exposure of amateurs is similarly unacceptable at the worst case level and could be refined by data from the skin penetration study. In addition, comparison with the 90 d NOAEL is considered particularly conservative when amateur use is expected to result in a very short-term and infrequent exposure. However, it has already been recommended that amateur use of products containing chlorothalonil be revoked because of the potential for skin sensitisation. 4.2.3.3 Risk Management - Professional Users Hooded air-fed respiratory protective equipment with a nominal protection factor of 50 or more is needed for the sprayer, and will also protect potentially exposed face and neck skin from contact. Other workers need for RPE should be determined by the employer through the workplace COSHH assessment. With regard to skin effects, it is assumed that if contact occurs then operators will be at risk. Workers should have a default set of protective equipment, which would include coveralls of a contrasting colour to the product being applied, worn beneath a disposable hooded coverall; protective gloves, and impervious footwear that protects the lower leg. To mitigate top-end exposure to hands, the protective gloves should be discarded at the end of each antifouling spray job (i.e. the gloves worn for one or two days only).

36

4.2.4 EXPOSURE ASSESSMENT - ANTIFOULANT BRUSH OR ROLLER PAINTING The following data are taken from guidance document HSE (2000) and Garrod et al (2000). No data have been received from industry and there are no data directly related to professional users. 4.2.4.1 Patterns of Use in Painting Operations - Amateurs and Professional (Chandlers) Users The HSEs information is that the work is seasonal (springtime), and may be performed on a slip-way, hard-standing or chandler's yard. Boat owners comprise the great majority of users, with a very small proportion of boats being treated with antifoulant by chandlers. The boat is cleaned with a high-pressure water-jet and may be scraped. The antifouling is applied using paint brush or paint roller. Rarely are more than two coats applied. Applying antifoulant employs no more than 2 persons per vessel. While amateur products have been approved for spraying, there is no information to indicate that this happens. Nor is there any information on the use of aerosol spray packaged antifoulant products for spot usage. The quantity of antifouling used per application session was found to range between 1.5 and 5 litres (median 4 litres; these were all copper-based products). Normally, 2 coats were applied to boat surfaces, which were found to range between 14 and 30 m2 (median 20 m2). Between 0.09 and 0.27 litres were applied per square metre (median 0.22 l m-2), at a work rate between 2.3 and 7.5 minutes per square metre (median 4.4 min m-2). The Environment Agency report has some comparative data on active substance and product usage. It indicated that the average boat size was around 30 ft, with motor boats (about 25% of all boats) generally larger than sailing boats (about 75% of all boats). Fouling was removed by pressure washing, and exhausted antifoulant removed with abrasive and/or a stripping preparation. Most boat owners applied antifouling themselves using a paint roller, annually. Around 15% of owners applied antifouling less than annually. The quantities applied ranged from 0.1 to 0.3 litres / square foot of boat. Antifouling took place mostly on hard standing or in a boat park. Chandlers were found generally to sell more than 23 products: 163 chandlers belonged to the British Marine Industries Federation, accounting for about 70% of all chandlers. Antifoulant was sold mostly between February and May. Amateurs normally apply antifouling products over one day or two consecutive days, normally during fine weather. The application time was found in the HSE survey to range between 35 and 112 minutes (median 90 minutes). 4.2.4.2 Work Clothing Work clothing worn by amateurs provides some degree of protection. While clothing will tend to retain liquids and dusts to some extent, there will also be run-off. Amateurs were found normally to wear coveralls and cloth gloves for painting. However, anecdotal evidence suggests that antifoulant application wearing minimal clothing is not uncommon. It is expected that chandlers would wear work clothing and protective gloves. Certain clothing may be desirable to mitigate general risks to health and safety, such as skin contact with solvent-based products. In such cases, gloves may be specified as a precautionary measure for amateurs to ensure that such general risks are minimised. In 37

addition, certain risk phrases derived through the CHIP (Chemicals (Hazard Information and Packaging for Supply) Regulations) process carry mandatory safety phrases and these will appear on a label even though there may be negligible risk to amateurs using the product. While amateurs may wear clothing such as coveralls or a long sleeved shirt and long trousers to apply non-agricultural pesticides, this cannot be assured, so may not be assumed for risk assessment purposes. Where there is no better information, default values for penetration to the skin are proposed at 5% when wearing clothes that provide some degree of protection (professionals at all times) and at 50 % for the realistic worst cases i.e. when minimal clothing is worn (by amateurs). The realistic worst case for amateurs also assumes that no gloves are worn. 4.2.4.3 Exposure Data for Painting Operations Detailed calculations are presented in Appendix 1, Tables 1.8 to 1.10. The very sparse exposure data relate to nine amateurs brushing and rolling their own vessels, with the boat on a cradle, trailer or sling. All but one of the painting jobs were outdoors, with both brush and roller used in most cases. There were nine data for potential dermal exposure, seven data for exposure inside gloves and two data for exposure to bare hands. Of the nine subjects, only four showed any exposure by inhalation. About half of the potential dermal exposure was to the legs. 4.2.5 SYSTEMIC EXPOSURES IN PAINTING OPERATIONS 4.2.5.1 Calculations The data sets and exposure calculations are set out in Appendix 1, Tables 1.8 - 1.10. Assumptions: 5 % active substance in the product 10 % dermal penetration. 5% or 50 % (amateur worst case) clothing penetration For professional application of antifouling products by brush and roller, calculations are presented in Table 4.10 with 5 % clothing penetration for workwear as explained in Section 4.2.4.2 of the main document. Calculations for amateurs are summarised in Table 4.11 and are presented for clothing penetration at 5 % and 50 %, as also explained in Section 4.2.4.2 of the main document. A summary of Tables 4.10 - 4.11 and resultant TER values is given in Table 4.12.

38

Table 4.10

[1.5 hour job; 5% clothing penetration; 1020 mg h-1 central tendency, 6480 mg h-1 95th percentile; 31 (median) & 1100 (worst case) mg h-1 of product in-glove]

Contact and Systemic Exposure to Chlorothalonil for Professionals (Chandlers): Application by Brush and Roller Exposure item Central Worst Case Tendency Amount of product in contact with skin (mg d-1) 123 2160

Percentage of active substance in product (%) Amount of chlorothalonil in contact with skin (mg d-1) Dermal absorption value (%) Systemic exposure to chlorothalonil via dermal route (mg d-1) Intake of product by inhalation (mg d )
-1

5 6.15 10 0.62 0.04 0.002 0.01

5 108 10 10.8 0.21 0.011 0.18

[1.5 hour job, 1.25 m3 h-1 inhaled volume, central tendency 0.04 mg m-3, realistic worst case 0.11 mg m-3, of product]

Amount of chlorothalonil inhaled (mg d-1)

Total systemic exposure for 60 kg operator (mg kg-1 d-1)

Table 4.11

Contact and Systemic Exposure to Chlorothalonil for Amateurs: Application by Brush and Roller Exposure item Central Worst Case Tendency Amount of product in contact with skin (mg d-1) 123 11500
[1.5 hour job; 5% central tendency clothing penetration, 50% worst case - minimal clothing; 1020 mg h-1 central tendency, 6480 mg h-1 95th percentile; 31 (median) & 4400 (worst case - no gloves) mg h-1 of product on hand ]

Percentage of active substance in product (%) Amount of chlorothalonil in contact with skin (mg d-1) Dermal absorption value (%) Systemic exposure to chlorothalonil via dermal route (mg d-1) Intake of product by inhalation (mg d-1)
[1.5 hour job, 1.25 m3 h-1 inhaled volume, central tendency 0.04 mg m-3, realistic worst case 0.11 mg m-3 of product]

5 6.15 10 0.62 0.04 0.002 0.01

5 575 10 57.5 0.21 0.011 0.96

Amount of chlorothalonil inhaled (mg d-1)

Total systemic exposure for 60 kg person (mg kg-1 d-1)

39

4.2.5.2 Assessment of Risk to Exposure by Brush or Roller Table 4.12 User A summary of Tables 4.11 - 4.12 and resultant TER values Worst Case TER

Central TER Tendency Professional users (NOAEL 3 mg kg-1 d-1) 5 % clothing penetration Chandler 0.01 300 Amateur users 5 % and 50 % clothing penetration Amateur 0.01 300

0.18

16

0.96

Systemic exposure of professional chandlers and amateurs is considered acceptable at the central tendency but gives cause for concern at the worst case level. As explained for spray operations, a better estimate of skin penetration of chlorothalonil from antifoulants would enable the risk assessment to be refined. Comparison with the 90 d NOAEL is again considered particularly conservative for amateur use, which is expected to result in a very short-term and infrequent exposure. However, it has already been recommended that amateur use of products containing chlorothalonil be revoked because of the potential for skin sensitisation. 4.2.5.3 Risk Management - Professional Users Professionals (e.g. chandlers) should wear a standard of personal protective equipment equivalent to that used by other professionals; coveralls of a contrasting colour to the product being applied, worn beneath a disposable hooded coverall with gloves and impervious footwear that protects the lower leg. 4.2.6 INHALATION EXPOSURE 4.2.6.1 Professional Users Central tendency In air conc. product without RPE (mg m-3) 6 0.6 0.8 0.02 In air conc. product with RPE (mg m-3) 0.12 N/A N/A N/A In air conc. In air conc. chlorothalonil chlorothaloni without RPE l with RPE (mg m-3) (mg m-3) 0.3 0.006 0.03 0.04 0.001 N/A N/A N/A TER

Sprayer Potmen Others Brush/roller

~18,300 (with RPE) ~3,600 ~2,750 110,000

40

Worst Case In air conc. product without RPE (mg m-3) 64.6 3.84 4.8 0.11 In air conc. product with RPE (mg m-3) 1.29 N/A N/A N/A In air conc. In air conc. chlorothalonil chlorothaloni without RPE l with RPE (mg m-3) (mg m-3) 3.23 0.06 0.19 0.24 0.006 N/A N/A N/A TER

Sprayer Potmen Others Brush/roller

~1,800 (with RPE) 579 458 18,333

4.2.6.2 Amateur Users Central Tendency In air conc. product without RPE (mg m-3) Sprayer 6 Brush/roller 0.02 Worst Case In air conc. product without RPE (mg m-3) 64.6 0.11 In air conc. chlorothalonil without RPE (mg m-3) 3.23 0.006 TER In air conc. chlorothalonil without RPE (mg m-3) 0.3 0.001 TER

367 110,000

Sprayer Brush/roller

34 18,333

The inhalation exposure of amateurs during spray application gives some cause for concern. However, it is already recommended that amateur uses of chlorothalonil-containing antifouling products be revoked on the grounds of skin sensitisation potential. 4.2.7 EXPOSURE AND RISK ASSESSMENT FOR BYSTANDERS No data are available for consumer exposure to antifouling products. It is considered that bystander exposure to antifouling products applied to the hull of large commercial vessels will be lower than for other workers, since spraying operations are generally avoided by others in the dry-dock, vessels do not emit chlorothalonil once antifoulant has settled on the ship surface and contact with wet surfaces by third parties is unlikely. For amateur applications, passers by within a congested yard could contact the hulls of freshly treated boats. There is a very low risk of skin sensitisation for bystanders if such exposure were repeated.

41

4.2.8 CONCLUSIONS The use of antifouling products containing chlorothalonil by professional operators is considered acceptable if operators wear air-fed RPE and coveralls of a contrasting colour to the product being applied, underneath a disposable coverall with hood, gloves and impervious footwear that protects the lower leg. To mitigate hand exposure, protective gloves should be replaced regularly, for example following antifoulant applications to a ship. Calculation of TERs suggests that this level of PPE would allow a large margin of safety against systemic effects. It is considered that potential exposure to amateurs to chlorothalonil by all methods is unacceptable based on the risk of skin sensitisation and that all amateur approvals for chlorothalonil should be revoked. No data are available for bystander exposure to antifouling products. However, there is minimal risk of skin sensitisation for bystanders.

4.3 RECOMMENDATIONS AND DATA REQUIREMENTS

Ministers have agreed the following recommendations and data requirements: 1. All amateur uses of antifouling products containing chlorothalonil are to be revoked because information from humans indicates that the risk of skin sensitisation is unacceptable. Approvals for the professional use of antifouling products containing chlorothalonil at a maximum formulation concentration of 5 % w/w may be continued for professional use by brush, roller and spray, subject to the fulfillment of data requirements. A study of the dermal penetration of chlorothalonil from formulations representative of approved antifouling products, to be submitted within 1 year. This study may be carried out in vitro or in vivo. The protocol should be discussed with HSE prior to commencement. Professional operators (sprayers) exposed to antifouling products containing chlorothalonil must wear RPE. Appropriate RPE includes air-fed respiratory equipment with combined protective helmet and visor to protect the skin of the head and neck. Impairment of vision should be avoided. For workers other than sprayers in the vicinity of the spray plume, RPE of an equivalent standard to FFP3 should be worn. For other workers, the need for RPE should be informed by a COSHH assessment. Unprotected persons should be kept out of treatment areas to minimise the risks posed by exposure to this compound. All professional operators exposed to antifouling products containing chlorothalonil should wear a disposable coverall with hood (providing head protection) and a second overall beneath this coverall of a contrasting colour to the antifouling product being applied. All bare skin should be covered. The disposable coverall should normally be used for no more than one spraying session. The second overall should be changed regularly and whenever product break-through has been detected.

2.

3.

4.

5.

6.

42

7.

Professional operators working with antifouling products containing chlorothalonil should wear impermeable gloves of a type recommended by the antifouling manufacturer as suitable for use with the formulation. These gloves should be changed regularly, e.g. after one or two days use. Operators should wear impermeable (and non-slip) footwear that protects the lower leg.

The following approval conditions are to appear on products' Notice of Approval and Schedules and they should be reflected on product labels using the following precautionary phrases: WEAR SUITABLE PROTECTIVE CLOTHING (COVERALLS OF A CONTRASTING COLOUR TO THE PRODUCT BEING APPLIED, BENEATH A DISPOSABLE COVERALL WITH HOOD), SUITABLE GLOVES, AND IMPERVIOUS FOOTWEAR THAT PROTECTS THE LOWER LEG. DISPOSE OF PROTECTIVE GLOVES AFTER USE If the product is to be applied by spray: UNPROTECTED PERSONS SHOULD BE KEPT AWAY FROM TREATMENT AREAS DO NOT BREATHE SPRAY MIST. WEAR SUITABLE RESPIRATORY EQUIPMENT SUCH AS AIR-FED RESPIRATORY EQUIPMENT WITH COMBINED PROTECTIVE HELMET AND VISOR WHEN SPRAYING WEAR SUITABLE RESPIRATORY EQUIPMENT SUCH AS FFP3 (OR AN EQUIVALENT STANDARD) WHEN WORKING IN THE VICINITY OF THE SPRAY PLUME.

43

ENVIRONMENTAL FATE AND BEHAVIOUR

The use of chlorothalonil as an antifoulant on vessels and in aquaculture was first reviewed by Committees in 1996. The environmental fate and behaviour data requirements were set as; a) b) c) d) e) Data on leaching rate of the active substance from surface(s) treated with the formulation. Adsorption and desorption properties of the active substance in soil or sediment as appropriate. Inhibition of microbial activity. Abiotic degradation: Hydrolysis as a function of pH. A reasoned case demonstrating that chlorothalonil can be used without adverse effects to the environment.

With the exception of e), the required studies have been submitted in full, together with an additional five studies addressing bioaccumulation of chlorothalonil in two species of fish which were not requested. These have been fully evaluated and are summarised below. The company also submitted summary data which had been prepared in support of the review of chlorothalonil as an agricultural fungicide under the European Plant Protection Products Directive (PPPD) (Unpublished, 1995b). Unfortunately, because of the summarised format of this data, it has not been possible to evaluate these data in the absence of full study reports. Therefore, only brief details of the additional studies, where endpoints are considered pertinent to the use of chlorothalonil as an antifoulant, have been included in this section.

5.1 Metabolite nomenclature

Throughout the fate and behaviour evaluation several metabolites have been identified, and the code given to these have in some cases changed over time. For the purposes of this document, the various metabolite descriptors and chemical names have been collated and are presented for easy reference in Table 5.1. It should also be noted that chlorothalonil is also coded as DAC-, DS- or SDS- 2787.

44

Table 5.1:

Structural and Identification Numbers of Potential Chlorothalonil Metabolites in the Environment

R1 R6 R2

R5

R3

R4

ID Number DAC-3701 DS-3701 SDS-3701 DAC-19221 DS-19221 SDS-19221 SDS-475231 SDS-47525 SDS-46851 SDS-67042 SDS-67042 sulfoxide SDS-13353 SDS-66382 SDS-66432

Chemical name 4-hydroxy-2,5,6trichloroisophthalonit rile 3-cyano-2,4,5,6tetrachlorobenzamide 3-Cyano-2,5,6trichlorobenzamide 3-Cyano-6-hydroxy2,4,5trichlorobenzamide 3-Carbamyl-1,2,4,5trichlorobenzoic acid 5-cyano-4,6,7trichloro-2H-1,2benzisothiazol-3-one 5-cyano-4,6,7trichloro-2H-1,2benzisothiazol-3-one sulfoxide 2,5,6-trichloro4-(thio) isophthalonitrile intermediate

R1 CN CONH2 CONH2 CONH2 CO2H

R2 Cl Cl Cl Cl Cl

R3 CN CN CN CN CO NH2 Cl Cl

R4 OH Cl Cl Cl Cl Cl Cl

R5 Cl Cl H Cl Cl CN CN

R6 Cl Cl Cl OH H Cl Cl

-CONHS-CONHSO-

CN CN

Cl Cl Cl

CN CN CN

SH GLUT
2

Cl Cl Cl

Cl Cl GLUT

2,5,6-trichloro-4CN (glutathione-5-yl) isophthalonitrile 1 SDS-47525 is the isomer of SDS-47523. 45

GLUT

GLUT refers to the glutathione conjugate.

5.2

HYDROLYSIS

In 1976, a study investigating the hydrolysis of chlorothalonil (purity 99.3 %/14C 99.8 %) and the known metabolite 4-hydroxy-2,5,6-trichloroisophthalonitrile (DAC-3701, purity 98.2 %/ 14 C 98.0 %) was carried out. The study was not carried out to specific guidelines or to GLP, and was conducted in three parts; test I, II and III. For each test, a 500 ml glass-stoppered flasks was prepared with the appropriate sterile buffer solutions (pH 5, 7 and 9) at nominal concentrations of chlorothalonil or DAC-3701. For test I samples; sterile buffer solutions were prepared at nominal concentrations of 0.50 mg l-1 chlorothalonil (pH 5 and 7) and 0.52 mg l-1 14C-chlorothalonil (pH 9 only). For test II samples; a nominal concentration of 1.5 mg l-1 14C-chlorothalonil was prepared for each of the three buffer solutions tested (pH 5, 7 and 9). And, for test III samples; solutions were prepared using a mixture of non-radiolabelled and radiolabelled 14C-DAC-3701 at a nominal concentration of 1000 mg l-1 for pHs 5, 7 and 9. In all three tests, once prepared, the flasks were sealed and covered with aluminium foil, shaken for two h, and stored at room temperature in the dark for periods of up to 89 d. Immediately prior to sampling, all test samples were shaken on an automatic shaker for 30 minutes. Samples were periodically removed from test flasks, acidified (sulfuric acid), extracted (isopropyl ether) and chlorothalonil levels analysed by gas chromatography (GC) with an electron capture detector (ECD). The formation and partitioning of water-soluble hydrolysis products was monitored by liquid scintillation counting (LSC). Further characterisation of radioactivity partitioned into the organic phase (ether extract) was accomplished by thin layer chromatographic (TLC) analysis. Loss by volatilisation was also determined by adding Aquasol to the test flasks followed by analysis using a Searle Analytical Ambient Temperature Counter. No further details were give regarding this method. Analysis of test I samples, by GC analysis indicated that the initial concentrations were 86 and 77 % of the nominal 0.50 and 0.52 mg ai l-1 levels. (This was postulated by the study author to have been due to either sampling techniques or adherence of the compound to glassware.) After 49 d exposure, under acidic (pH 5) or neutral conditions (pH 7), chlorothalonil concentrations remained unchanged. However, at pH 9 a significant (80 %) decrease in chlorothalonil concentration was reported. LSC analysis of the pH 9 sample extracts demonstrated that 98.7 % of the applied radioactivity (AR) was partitioned to the organic phase. Additional analysis of the pH 9 sample extracts using GC, TLC and mass spectral techniques identified two metabolites; DAC-3701 and 3-cyano-2,4,5,6tetrachlorobenzamide (DS-19221). Further quantitative analysis by TLC combined with LSC, demonstrated that 24 % of the applied chlorothalonil remained intact; 22 % of the applied ai had degraded to DAC-3701 and 54 % to DS-19221. These results supported earlier GC data which showed 20 % of the applied chlorothalonil remained intact after 89 d at pH 9. The hydrolysis rate of chlorothalonil was determined by computerised least squares analysis (plotted as % chlorothalonil remaining vs. time). The resultant gradient of -0.0183 indicated that the hydrolysis of chlorothalonil followed first order kinetics.

46

The initial nominal concentration of test II solutions (1.5 mg ai l-1) exceeded the water solubility of the compound (0.6 mg l-1), and only 0.95 mg l-1 was measured at test initiation. At pH 5 and 7, samples exposed for 72 d demonstrated slight increases in the solubility with time (16.9 and 17.7 % respectively). Ninety-nine and 98.2 % of the AR partitioned to the organic extracts at pH 5 and 7 respectively, of which 97.2 and 95.5 % was identified by TLC and LSC analysis as chlorothalonil. At pH 9 the water solubility of the 14C-residues was observed to increase significantly from 0.95 to 1.43 mg l-1, representing a 50.5 % increase. The increased solubility over time was explained by the author to be due to the increased production of DAC-3701 and DS-19221, which are more soluble in water than chlorothalonil. After 72 d, only 36.4 % of the AR remained as chlorothalonil, 48.9 % AR had degraded to DS-19221, 11.3 % AR had degraded to DAC-3701. A further 5.1 % AR was present as unidentified, water-soluble metabolites. In test III samples, slight increases in water solubility were demonstrated after 72 d, but DAC-3701 remained at 98.5 % of the AR at all pH levels throughout the study. The remaining 1.51 % AR was reported as unidentified water-soluble products. From the data presented, it may be concluded that chlorothalonil was resistant to hydrolysis at neutral and acidic conditions. However, at pH 9 hydrolysis to DAC-3701 and DS-19221 occurred. The latter compound was considered to have been the primary hydrolysis product since 55 % of the applied chlorothalonil was detected as DS-19221 after 89 d. Whereas, only 22 % of the applied chlorothalonil degraded to DAC-3701 after a similar exposure period. Throughout the study excellent agreement between GC and radiotracer data was obtained, with no losses due to volatility or the formation of volatile hydrolysis products. The study author suggested a hydrolysis pathway for chlorothalonil at pH 9 (presented in Figure 5.1). The known metabolite DAC-3701 exhibited extreme stability and resistance to hydrolysis when stored in aqueous solutions at pH levels of 5, 7 and 9, (Unpublished, 1976). The submitting company also provided a copy of the above study in its published form (Szalkowski and Stallard, 1977). In the published version, the reaction slope was recalculated by plotting 'log % remaining chlorothalonil vs time', and a gradient of -0.0079 reported. From these data, chlorothalonil was calculated to have declined at a daily rate of 1.8 %, and the hydrolysis half-life of chlorothalonil at pH 9 was calculated as 38 d.

47

CN Cl Cl

Cl Cl

CN

Chlorothalonil tetrachlorisopthalonitrile

Hydrolysis pH 9
CN Cl Cl Cl

O C

NH2

+
Cl OH CN

Cl

Cl Cl

CN

4-hydroxy-2,5,6-trichloroisophthalonitrile DAC-3701

3-cyano-2,4,5,6-tetrachlorobenzamide DS-19221

Figure 5.1: Proposed Hydrolysis Pathway of Chlorothalonil Exposed to pH 9

5.3

AQUEOUS PHOTOLYSIS

A brief summary of a study addressing aquatic photolysis was submitted. Sterile 14C-chlorothalonil solutions were exposed, continuously, to 118 h of artificial sunlight over an 11 day period (equivalent to 30 d of 12 h sunlight d-1). Both test and dark control solutions were maintained at 25 oC and pH 5. At the end of the exposure period, samples were acidified, extracted and analysed by HPLC. The data were transformed using a least squares regression fit of time versus log concentration. This resulted in a straight line (r = 0.994), from which a half-life of 64.7 d was calculated, indicating that aqueous photolysis would not represent the major degradation route for chlorothalonil in the environment. Although the amounts were not given, it was stated that the major metabolite was identified as SDS-3701,

48

and several minor unidentified metabolites both polar organic-soluble and water-soluble compounds (Unpublished, 1995b).

5.4

PHOTOLYSIS ON SOIL SURFACE

A brief summary of a study addressing photolytic stability of chlorothalonil in soil and the major soil metabolite SDS-3701 was submitted. Soil/glass TLC plates were prepared using five different soil types (two silty loam, and three silty clay loam soils), and spotted with 14Cchlorothalonil/SDS-3701. The plates were exposed to an artificial light source equivalent to 168 d of 12 h sunlight d-1, following which the plates were developed by TLC. Extractability was > 97 %, with 97 and 84 % recovery of AR for the chlorothalonil and SDS-3701 samples respectively. Therefore, both compounds were demonstrated to be stable to photolysis under the conditions tested (Unpublished, 1995b).

5.5

INHIBITION OF SOIL MICROBIAL PROCESSES

Three soil microbial inhibition studies (submitted to address data requirement set for inhibition to microbial activity) were conducted between 1980 and 1981, these were not carried out to recognised guidelines but to established in-house protocols. GLP was not claimed, but each study included a quality assurance statement. The results of these studies are presented in a simple format since no gross effects were observed, and no read across to effects on sewage sludge or sediment micro-organisms can be made. The preparation for each study was very similar and is outlined below. In all three studies the same two standard US soils were used; sandy loam and clay loam soil (characteristics in Table 5.2.), which were fortified with chlorothalonil at nominal levels of 0 (blank), 2.5 (recommended field application rate) and 25 mg ai kg-1 soil (10 x field rate). For each soil type at each fortification level, sufficient amber bottles were prepared so as to provide one sample set for each time period of the testing schedule for each substrate tested and all test vessels were stored at 22 1C. Each sample set consisted of triplicate soil samples for each soil type at each nominal concentration. The homogeneity and the experimental concentration of chlorothalonil in each soil was verified using GC analysis. Studies I and II were carried out under aerobic conditions only, which were maintained by flushing each test vessel three times each week with carbon dioxide-free, water-saturated air. For study II, half of the test vessels were also maintained under anaerobic conditions by flushing in a similar manner to the aerobic flasks but with water-saturated nitrogen gas.

49

Table 5.2: Soil Characteristics

Soil C.E.C. Classification (meq/100g) Clay loam Sandy loam 16.5 2.6

pH

Organic matter

Percentage sand silt

clay

Study I (1980) 3.9 6.68 0.06 1.7 6.85 0.14 Study II & III (1981) 3.6 6.83 0.02 1.3 7.03 0.07

15.8 78.7

57.7 16.9

26.5 4.5

Clay loam Sandy loam

16.2 2.1

19.7 77.4

50.4 12.7

29.8 9.8

The validity of each study was determined by assessing the control soils at test initiation, midpoint and conclusion. Firstly, measurements of microbial density and diversity were taken to ensure that a large microbial population was present and that no one population had predominated. It was reported, that for each study, a high level of microbial activity and diversity for each soil type was present. Further to this, substrate degradation was assessed in all control soils. Again it was reported that for each study the level and reproducibility of substrate degradation within the control soils was consistently good. Based upon these results, the test systems used were judged to be reliable and valid for evaluating the effect of chlorothalonil upon soil micro-organisms and their processes. In all three studies, analysis of variance (ANOVA) and Duncans multiple range test, were used to assess the statistical significance of the data. 5.5.1 DEGRADATION OF PROTEIN, PECTIN, CELLULOSE AND STARCH BY SOIL MICRO-ORGANISMS In 1980, a study to determine the effects of chlorothalonil upon the degradation of protein, pectin, cellulose and starch by soil micro-organisms. The mean measured concentrations of chlorothalonil in the clay loam samples were reported as 2.42 and 22.62 mg ai kg-1 soil, and the sandy loam samples reported as 1.95 and 15.71 mg ai kg-1 soil. On days 0 (immediately following sample fortification), 3, 7, 14, 21, 28 and 56; 100 mg of 14C-substrate (either protein, pectin, cellulose or starch) was added to each respective sample set. The samples were then purged with carbon dioxide-free dry air; 24, 48, 72 and 96 h after substrate addition, the effluent gas from each sample trapped and 14CO2 immediately quantified using LSC techniques. The effect of chlorothalonil upon the microbial utilisation of protein, pectin, cellulose and starch was then evaluated by comparing the percentage of AR degraded to 14CO2 from the control vessels with that from the fortified soil samples.

50

Generally, any effects of chlorothalonil upon the utilisation of protein, pectin, and cellulose were subtle or isolated in both soils and did not persist beyond seven days. A stimulatory effect upon the utilisation of starch was reported throughout the study for sandy loam soil, which appeared to be positively correlated to chlorothalonil concentration. In the clay loam soil this effect was only observed up to day 14, but was also positively correlated to chlorothalonil concentration. Stimulation of starch utilisation by soil was not considered by the study author to be biologically detrimental to the soil environment. Therefore, under the conditions of this study, chlorothalonil was not considered to have adversely affected the soil micro-organisms responsible for the microbial processes tested (Unpublished, 1980a). 5.5.2 NON-SYMBIOTIC NITROGEN FIXATION BY SOIL MICRO-ORGANISMS

In 1981, a study to determine the effect of chlorothalonil upon non-symbiotic nitrogen fixing soil micro-organisms was assessed. For this study the soil samples (mean measured concentrations; 1.8 and 19.0 mg ai kg-1 soil for clay loam and 1.6 and 19.9 mg ai kg-1 soil for sandy loam), were stored under both aerobic and anaerobic conditions. On days 0 (immediately after fortification), 3, 7, 14, 21, 28 and 56 dextrose (unknown quantity) was added to the sample sets which were purged with carbon dioxide-free air or nitrogen gas as appropriate. Seven days after dextrose addition, acetylene (99.6 % purity) was introduced to each sample. At intervals of 0, 6, 24 and 48 h following acetylene addition, samples were analysed by gas liquid chromatography (GLC) for acetylene reduction (formation of ethylene). The effect of chlorothalonil upon aerobic and anaerobic nitrogen fixation was then evaluated by comparing the reduction of acetylene to ethylene in the control and fortified soil samples. The results suggested that although dose specific, isolated effects were reported, chlorothalonil had no significant or long-term effects on aerobic or anaerobic nitrogen fixation, in either soil type tested (Unpublished, 1981a). 5.5.3 NITROGEN TRANSFORMATION BY SOIL MICRO-ORGANISMS

In 1981, a study to determine the effect of chlorothalonil upon nitrogen transformation was assessed using fortified soils . The reported mean measured concentrations of chlorothalonil at the start of the test in sandy loam soil were 1.6 and 19.9 mg ai kg-1 soil, and in the clay loam soil were 1.8 and 19.0 mg ai kg-1 soil. On days 0 (immediately after fortification), 3, 7, 14, 21, 28 and 56, ammonium (amount not specified) was added to soil samples which were then maintained at approximately 22 C for 2 - 10 d prior to the assay. The effect of chlorothalonil upon nitrogen transformation was evaluated by comparing the concentrations of ammonium, nitrate and nitrite in the control against those for the fortified soil samples, using specific ion electrode techniques. From the resulted, it was concluded that the effects on nitrogen transformation in sandy loam and clay loam soils fortified at the lower dose (1.6 and 1.8 mg ai kg-1 respectively) were subtle and isolated with no observed trends.

51

At the higher dose, clay loam soil fortified with 19.0 mg ai kg-1 soil observed sporadic inhibitory effects on nitrification, therefore, no clear trends could be made. However, in sandy loam soils (19.9 mg ai kg-1 soil), ammonium levels were reported to have been significantly greater than those of the controls at all sample times except for day 0. The actual percentage increase in levels of ammonium decreased over the course of the study from 401.76 % on day 3, to 14.35 % by day 56. In summary, there was a consistent inhibitory effect on nitrogen transformation at the higher chlorothalonil fortification level in sandy loam soil, but the results suggested recovery of the soil microbial system was occurring (Unpublished, 1981b).

5.6 5.6.1

DEGRADATION IN SOIL
AEROBIC METABOLISM

A company summary of three studies addressing degradation in soil under aerobic conditions was submitted. Study I: Four soil types (see Table 5.3 for characteristics) radiolabelled with chlorothalonil were stored in the dark under sterile and non-sterile conditions at 25 C. Table 5.3: Soil Characteristics % Soil Classification (No) Silty clay loam (1) Peat loam (2) Sandy loam (3) Sandy loam (4) pH 5.1 7 8 6 Organic matter 2.31 7.61 1.6 3.2 sand 0.6 29.7 54 62.2 silt 75.6 50 25 31 clay 25.8 20.3 19.5 6.8
14

C-

Duplicate samples were analysed for chlorothalonil and metabolites on days 0, 7, 16, 22, 31, 60 and 90 and the results are presented in Table 5.4. Table 5.4: Percentage Remaining 14C-chlorothalonil, and Calculated Half-lives % total 14C-activity remaining Soil Type (No) Peat loam (2) Sandy loam (3) Sandy loam (4)
1

0 97.6 92.2

7 80.9 61.6 44.8

16 67.2 42.4 36.4 32.2

22 62.4 34.3 30.7 21.1

31 46.5 23.9 26 11.2

60 51.1 14.4 12.8 10.8

90 33.3 6 4.6 4.8

T1 (d) 36.5 14.7 12.8 10.3

Silty clay loam (1) 94.8

100.2 60.2

- K = (2.303/t)log(a/a-x)

52

From these data it was concluded that chlorothalonil underwent rapid degradation in four different soils, and although it was stated that the pattern of chlorothalonil degradation was similar in all four soil types, only one set of results for the sandy loam (4) was given. Following 90 d incubation, the majority of the breakdown products were characterised as unextractable residues (62.8 % AR), SDS-3701 (13.5 % AR), and unidentified water-soluble residues (10.5 % AR). SDS-19221 was also identified but remained < 10 % throughout the study. It should be noted that for there is some confusion in the summary data between sandy loam (3) and (4) in the results tables given. Study II: The bound residues which resulted from Study I (90 d aerobic incubation with 14Cchlorothalonil) were characterised according to the EPA guideline for Registering Pesticides in US (Federal Register 40 (123): 26802-26928 June 25 1975). The results from this study are given in Table 5.5. Table 5.5: Distribution of the Bound 14C-residues in Soil % Total Radioactive Residue (TRR) o Soil Type (N ) Humin Humus Fulvic II III IV 11.2 3.8 10.3 Silty clay loam (1) Peat loam (2) Sandy loam (3) Sandy loam (4) 15.9 18.5 12.8 10.1* 2.8 8.6 10.3 11.9 14

[All mean of two replicates, except for* which was not duplicated] Only a small fraction of the bound Fulvic IV 14C-residues (approx. 4 - 8 % of TRR) were characterised (tentatively) as SDS-3701 (Unpublished, 1995b). Study III: A large scale aerobic soil metabolism study was carried out in order to obtain sufficient 14C-residues to allow for full characterisation. For this study, a sandy soil (no properties given) was fortified with 14C-chlorothalonil at 10 mg kg-1 soil week-1 for 13 weeks. The treated soil was then incubated in the dark, at 24 1 C until approximately 85 % of the applied 14C-chlorothalonil had degraded. The soil was subject to a variety of extraction techniques which resulted in 95.3 % recovery of 14C-residues which were analysed by TLC methods to give the results presented in Table 5.6.

53

Table 5.6: Quantitative Distribution of 14C-residues in Sandy Loam Soil Residue Bound I (SDS-3701 Chlorothalonil (SDS-2787) II (SDS-19221) Unknown III Unknown IV Unknown V % (TRR) 26.8 22.3 15.5 10.4 4.3 3.8 3.2

Unknown metabolites III, IV and V represent the polar water-soluble residue fraction. These were separated, further purified and analysed by mass, infrared and NMR spectroscopy which identified the metabolites as; III SDS-46851 3-Carbamyl-1,2,4,5-trichlorobenzoic acid IV SDS-47525 3-Cyano-6-hydroxy-2,4,5-trichlorobenzamide V SDS-47524 3-Cyano-2,5,6-trichlorobenzamide. As a result of this study, the summary data details an additional assessment of the chlorothalonil degradation data (Study I), and reported that chlorothalonil underwent biphasic decay in non-sterile soil. From this new half-lives were calculated for chlorothalonil in soil (see Table 5.7), and for SDS-3701 the degradation half-life was quoted never to exceed 43 d. Table 5.7: Recalculated Chlorothalonil and SDS-3701 Half-lives Soil Type (No) Sample 1 Silty clay loam (1) 2 1 Peat loam (2) 2 1 Sandy loam (3) 2 1 Sandy loam (4) 2 Half-life (d) Chlorothalonil SDS-3701 30 11 36 40 13 33 15 43 13 20 13 16 8 8 11 6

54

5.6.2

ANAEROBIC METABOLISM

A study summary was submitted. The study used two soil types, silty loam (0.7 % organic matter - O.M[HSE1]) and sandy loam (3.5 % O.M), which were flooded and incubated under atmospheric nitrogen for 30 d. Following this 14C-chlorothalonil was added to the soils, which were then incubated for a further 60 d. The soils were maintained in the dark at 25 + 1 oC. Soil and water samples were taken on days 1, 3, 5, 10, 15, 30 and 60 d post 14C-chlorothalonil addition, extracted and analysed by LSC and HPLC. Recovery of 14C-residues were acceptable at > 85 % in both soil systems. The metabolites formed were the same as those identified in the previous aerobic soil metabolism studies, with the most significant metabolite identified as SDS-3701 (maximum 42.8 and 17.7 % TRR in silty and sandy loam soils respectively) (Unpublished, 1995b).

5.7

AQUATIC DEGRADATION

A summary of a study carried out to determine the rate of degradation and metabolic fate of chlorothalonil in fresh and saltwater-sediment systems under aerobic conditions was submitted. Both test systems, fresh and saltwater, were prepared using 9 parts water to 1 part sediment. Triplicate flasks, for each test system, were incubated in the dark for 3 d at 25 + 1 oC with constant agitation to ensure aerobicity. After this, 14C-chlorothalonil was added to two flasks, leaving the third as a control. All the flasks were then further incubated in the dark at 25 + 1 oC, and at appropriate time intervals (i.e. 1, 3, 6 & 12 h and 1, 3, 9, 16, 23 & 30 d) aliquots of sediment and water were removed and analysed. At each sampling point, the flasks were sufficiently agitated to ensure a homogeneous suspension was sampled. The treatment of the sample involved centrifugation, followed by a series of extraction techniques on both sediment and water fractions. The volume of water, pH and dissolved oxygen were monitored and maintained throughout the study. The results suggested that chlorothalonil was rapidly metabolised, with a T of < 2 h, in both fresh and saltwater-sediment systems. The 14C-residues could be divided into three fraction; organic soluble, polar water-soluble and bound (see Table 5.8). Table 5.8: Percentage Distribution of 14C-residues % of TRR Organic soluble Saltwater-sediment Freshwater-sediment 39 -68 56 - 96 Polar, water-soluble 1-9 3 - 15 Bound 28 - 62 4 - 22

Two major and three minor metabolites were formed in each system. Further analysis using HPLC or GC-MS identified these metabolites as;

55

1.

SDS-67042 (5-cyano-4,6,7-trichloro-2H-1,2-benzisothiazol-3-one) which reached a maximum level of 29.2% TRR on day 9 in saltwater, and 30.9 % TRR on day 1 in freshwater. SDS-67042 sulphoxide, which reached maximum levels of 12.1 and 16.5 % TRR on day 9 in salt and freshwater respectively. SDS-66432 (2,5,6-trichloro-4-(glutathione-5-yl)isophthalonitrile) which peaked at < 10 % TRR in both systems. SDS-13353 (2,5,6-trichloro-4-(thio)isophthalonitrile) which was always < 10 % TRR at all sample times in both systems tested. SDS-3701 (major soil metabolite) which never exceeded 10 % TRR throughout the study, in fact the maximum level recorded was 5.4 % TRR in the freshwater system.

2. 3. 4. 5.

Further acid hydrolysis of the polar water-soluble and bound 14C-residues from both systems tested released only small amounts of metabolites which were the same as those identified above. The conclusions from this study are that chlorothalonil was rapidly metabolised (T of < 2 h) in water-sediment systems, and that the pattern of metabolites was similar in fresh and saltwater systems. SDS-3701 which is a major soil metabolite was not a major metabolite under the conditions of this study. The breakdown of chlorothalonil was described in the report as microbial in origin, involving an attack on chlorothalonil by glutathione (or other sulfur species) and it was suggested that this occurred at the sediment interface. It was predicted that the two major metabolites would undergo further metabolism resulting in the formation of bound and polar water-soluble residues (Unpublished, 1995b). Numerous aquatic degradation studies were available from the public domain which have been summarised below. Davies (1988) carried out a series of experiments to investigate the route and rate of chlorothalonil disappearance in the aquatic environment at different temperatures with different stream substrates. The test systems consisted of natural water (pH 6.8 -7.1) or sterilised water (20 l) which was spiked with an acetone stock of chlorothalonil or 14Cradiolabelled chlorothalonil to give a final concentration of 20 mg ai l-1 and placed in glass aquaria housed in constant temperature rooms (5 and 15 C). Duplicate tanks were set as described in Table 3.2.14. At various time intervals over a period of 75 - 336 h, water, algal and fish samples were taken, extracted with hexane and analysed by GC-ECD (detection limit 0.001 g l-1). Radioanalysis of 14C-chlorothalonil was carried out using LSC with GC-ECD. The reported results are presented in Table 5.9.

56

Table 5.9: Degradation Half-lives of Chlorothalonil in Natural Stream Waters Water Temp Half-life origin C (h) Still water* Natural 5 150 Still water* Natural 15 80 Aerated water Distilled 15 106.3 Aerated water Natural 15 101.3 Aerated water Natural 15 107.3 Aerated water + rock + algae* Natural 5 13.9 Aerated water + rock + algae* Natural 15 7.7 Aerated water + rock + algae Natural 15 4.4 Aerated water + rock Natural 15 90.6 Aerated water + fish (Galaxias auratus) Natural 15 4.3 14 * indicates use of chlorothalonil, all other tests were performed using C-chlorothalonil. Conditions Temperature had a marked effect on the degradation half-life of chlorothalonil, a 10 C decrease in temperature doubled the reported half-lives, strongly supporting the authors theory that degradation was of a biological nature. The type of aerated water used in the tests had no effect on the rate of degradation, but did have a marked effect on the appearance rates of the polar metabolites. Distilled water resulted in an appearance half-life of 567.3 h compared to the metabolite appearance half-lives in natural waters of 161.3 - 172.1 h which suggested that the appearance of the metabolites was affected by biological biomass and/or solute concentration, whereas apparent degradation in distilled water may have been due to volatilisation (this was not discussed). Further investigations within the report highlighted that chlorothalonil was strongly associated with the suspended material in the stream water and a log Pow of 4.38 was determined. The presence of algae was seen to increase the removal of chlorothalonil from the water column, but did not increase the rate at which the polar metabolites appeared. From the analysis of the algal material a BCF of 270 was reported, which was substantially higher than the BCF of 18 which was reported for G. auratus. However, the rate at which chlorothalonil degraded was enhanced by a factor of 25 times in the presence of G. auratus, and the appearance rate of the metabolites increased by a factor of three. The author concluded that chlorothalonil was biodegraded at low concentrations, but that it was unlikely that biodegradation would play a major role in the fate of chlorothalonil in moderate to fast flowing streams where stripping by adsorption were likely to be the dominant factors. Walker et al., (1988) investigated the first-order biotic and abiotic degradation constants of 14 pesticides including chlorothalonil in sediment/water systems. The water and sediment used were taken from two US sites. Nominal test concentrations of 200 mg l-1 chlorothalonil were prepared and one litre transferred to duplicated Erlenmeyer flasks containing either; nonsterile water only, non-sterile sediment in water, sterile water only and sterile sediment in water. The flasks were then placed in an orbital incubator (150 rpm) and maintained at 25 C in the dark. The pH was measured every other day and remained within 0.2 units of the original pH (not stated). Duplicate water sub-samples were removed at regular intervals from each flask and the parent compound analysed by GC-ECD (recovery was > 85 %).

57

First-order degradation rates in sterile and non-sterile flasks were compared in the presence and absence of sediment to investigate the role of sediment in biodegradation. Using the reported rate constants HSE calculated the half-life of chlorothalonil for each of the four reported test conditions using the known formula T = 0.693/k1 (see Table 5.10). Table 5.10: Degradation Half-lives of Chlorothalonil in Estuarine Water and Sediment-water Test Systems Test medium Sediment/water Non-sterile Sterile Water only Non-sterile Sterile Half-life (d) 1.79* 5.03 8.13 10.2

* significantly faster (p 0.01) than all other rates reported. The presence of sediment in flasks enhanced the loss of chlorothalonil from the system under both sterile and non-sterile conditions. However, the significant difference between the degradation rates of the sterile and non-sterile sediment-water test systems suggested that biodegradation and not adsorption was the key process (all other factors such as sorption, volatility, photolysis and pH were the same). It should be noted that only parent compound was analysed for, therefore, the degradation pathway of chlorothalonil under the conditions tested was not addressed further. Gajbhiye et al., (1989) studied the persistence of chlorothalonil in water and sediment contained in PVC tubs. PVC tubs were prepared with a 8 kg of sandy loam soil (pH 8.2, organic carbon 0.64 %, CEC 15.9 meq/100g soil) overlaid with 40 l of water (no further details). The soil and water were mixed and left to equilibrate for 48 h prior to the addition of chlorothalonil directly to the water column. Two application rates equivalent to 1 and 2 kg ai ha-1 were used, and each application rate was applied to three replicate tubs. Periodic water and sediment sub-samples were taken, extracted using methylene chloride and acetone respectively, followed by GLC-ECD analysis. The initial concentrations in water 1 h after application were 0.465 and 1.074 mg ai l-1 for the 1 and 2 kg ai ha-1 application rates respectively. The subsequent loss from water was initially rapid with 39.1- 43.4 % of the applied chlorothalonil lost within 24 h, 72.4 - 77.8 % lost within 3 d and 97.4 - 97.7 % lost within 7 d. Further loss was slow with residues of 0.006 mg ai l-1 persisting up to 30 d. Analysis of the sediment showed that most of the chlorothalonil lost from the aquatic portion was primarily due to adsorption with levels of 0.21 and 0.31 mg ai kg-1 reported for sediment 24 h post application, increasing to 0.26 and 0.33 mg ai kg-1 sediment 3 d post application of 1 and 2 kg ai ha-1 respectively. After this time the concentrations within the sediment samples declined, and on day 30, less than 0.01 mg ai kg-1 sediment was reported. Therefore, under the conditions of this study, chlorothalonils primary route of removal from the water column was via adsorption to the sediment, where subsequent degradation was shown to have occurred. Metabolites were not analysed for, and no half-lives were calculated.

58

Callow and Willingham (1996), investigated the degradation of chlorothalonil over an eight week period using a bioassay developed with the ship-fouling diatom, Amphora coffeaformis. Degradation of chlorothalonil was measured in three different seawater media prepared from filtered natural seawater. The media used were; natural seawater (NSW), which contained a normal compliment of micro-organisms; sterile seawater (SSW), which had been autoclaved and biomass enhanced seawater (BSW), which contained added pre-cultured marine bacteria. Chlorothalonil (> 97 % pure) stock solutions were prepared in DMF (dimethyl formamide) and nominal test concentrations of 0 (control) and 2.0 mg ai l-1 for each media type was prepared. The selected test concentration corresponded to the previously established EC80 (growth) concentration for A. coffeaformis. Test solutions were then capped and stored at 25 C in the dark. After periods of 2, 4, 6 and 8 weeks test solutions were taken and concentrated cultures of A. coffeaformis were added with sufficient nutrient media to sustain the diatoms normal log phase growth. The test solutions were then stored in an illuminated orbital shaker at 20 C for 96 h, after which time chlorophyll a was extracted from the diatom sample and quantified. The results were expressed as mean percentage inhibition relative to the control, and significance was defined using 1 and 2-way analysis of variance (ANOVA). After 8 weeks exposure there was no loss of biocidal activity in the SSW test vessels, however, approximately 25 and 50 % of the biocidal activity was lost in the NSW and BSW treatments respectively. Therefore, the bioassay results suggested that no apparent abiotic degradation had taken place in the SSW, but biodegradation was apparent after 4 weeks and proceeded faster in the BSW (T 8 wk) medium compared to that of the NSW (T > 8 wk) medium. Bacterial counts demonstrated that the BSW had a greater number of bacteria present than NSW, and that chlorothalonil addition resulted in an initial decline in bacterial numbers for both media treatments. However, recovery was seen by the sixth week of the study. The authors concluded that this bioassay provided an indication of the potential degradability of chlorothalonil in the marine environment by assuming the concentration of chlorothalonil was fixed by its efficacy/toxicity to the test organism. It was also stated that the derived half-life value was not expected to be wholly comparable to chemically derived halflives but merely represent the time taken for half the bioactivity of the ai to have disappeared.

5.8

ADSORPTION AND DESORPTION

In 1982, a study was carried out to determine the adsorption and desorption of 14Cchlorothalonil (purity > 97 %) in four US soils. The study was not conducted to a recognised guideline or to GLP. However, the study was conducted to an established in-house protocol, which was submitted along with a quality assurance statement reporting that the study had been assessed as accurate and acceptable. Silty clay loam, silt, sand, and sandy loam soils (characteristics in Table 5.11.), were air-dried (24 h) and sieved prior to use. Nominal test solutions of 14C-chlorothalonil (0.5, 0.4, 0.2 and 0.1 g ml-1) and a reference compound 14C-DDT (0.05, 0.04, 0.02 and 0.01 g ml-1) were prepared in 0.03N CaSO4 (pH 7.0). The actual concentrations of test solutions were measured by LSC and were reported to have been 103.4 and 103.2 % of the nominal concentrations for 14C-chlorothalonil and 14C-DDT respectively.

59

Table 5.11: Characteristics of Three US Soil Types

Soil Classification C.E.C. (meq/100g) Organic matter Silty clay loam 25 3.2 Silt 10.2 0.7 Sand 1.7 0.6 Sandy loam 11.8 3.2

% sand 9 9 96 64.6

silt 63 83 0 29.4

clay 28 8 4 6

A preliminary test using a 0.5 g ml-1 solution of 14C-chlorothalonil was carried out to establish the equilibrium times for each soil. From this an 8 h equilibrium period was established for the silty soils and a 24 h equilibrium period for the sandy soils tested. For the adsorption phase, triplicate vessels of each soil type were prepared at each test concentration. The tubes were capped, hand-shaken, and placed in a shaker bath for the appropriate equilibration time. After equilibration, the samples were removed from the bath, centrifuged, and the radioactivity in the supernatant quantified by LSC. Immediately the after adsorption phase, a 4.0 ml portion of each supernatant was removed and a 4.0 ml portion of uncontaminated 0.03N CaSO4 added. Each sample was then shaken for an additional period equivalent to that of the adsorption phase, centrifuged and the radioactivity in the supernatant quantified. This desorption phase was repeated once more. The adsorption and desorption of the reference substance 14C-DDT was reported to have been within the limits set by the laboratory protocol for this study. The silt and clay loam soils both adsorbed approximately 90 % of the chlorothalonil in solution within 8 h. The sandy loam and sand soils adsorbed 84 and 47 % of the chlorothalonil in solution respectively, within 24 h. For each soil type the results were presented as linear isotherms, indicating that the adsorption of chlorothalonil was in accordance with the Freundlich adsorption isotherm equation. Linear regression analysis of the data was used to estimate K (adsorption constant) and 1/n (slope) for each soil (see Table 5.12). However, no definite correlation between adsorption and the percent organic matter could be established (i.e. the value Q).

60

Table 5.12: Adsorption, and Freundlich Isotherm Constants of 14C-chlorothalonil

Freundlich isotherm constants Soil type Silty Clay Loam Silt Sand Sandy Loam Equilibrium time 8h 8h 24 h 24 h Mean % Adsorption 93.7 1.1 93.4 1.2 57.1 5.3 85.8 1.0 % Organic matter 3.2 0.7 0.6 3.2 K 26 29 3 20 Koc* 3,300 14,000 1,300 1,600 1/n (slope) 0.7919 0.8334 0.7525 0.9441 Q 813 4,143 500 625

* not reported in original report but reported in the summary data (independent calculations by HSE confirm these are correct).

According to the Soil Survey and Land Research Council (SSLRC) classification for mobility in soil under the conditions tested, chlorothalonil was slightly mobile in all the soil types tested with the exception of silt in which it was classified immobile. Only a small portion of the adsorbed chlorothalonil was desorbed (see Table 5.13) and the rate of desorption was reported to have been independent of concentration. For the three strongly adsorbing soils, less than 7 % of the adsorbed chlorothalonil was desorbed with each desorption phase. Whereas, up to 28 % of the chlorothalonil adsorbed by sand was desorbed during one desorption phase. However, only 50 % of the media used in the adsorption step was removed and replaced with uncontaminated medium for each desorption step. Therefore, it is possible that desorption was underestimated especially with respect to the first desorption step. Table 5.13: Percentage Desorption of Applied 14C-chlorothalonil Mean Percentage ( SD) Soil Type Silty Clay Loam Silt Sand Sandy Loam Desorption 1 Desorption 2 2.0 0.2 1.9 0.1 10.4 0.9 4.7 0.3 2.6 0.3 2.8 0.4 24.2 4.5 6.2 0.5

Data from this study suggested that chlorothalonil would be adsorbed quite strongly to soils (and sediments) other than sand. However, because of the methodology undertaken in assessment of desorption, the capacity of chlorothalonil to leach from soils, or be remobilised from sediments has not been fully established (Unpublished, 1982). This study was also submitted in summary form, along with additional results from two studies which investigated the mobility of two soil metabolites, SDS-3701 and SDS-46851. Both metabolites were reported to have been more mobile than the parent compound, with reported Koc values ranging between 251 - 491 (SDS-3701) and 74 - 169 (SDS-46851). Under SSLRC classification scheme, SDS-3701 would be classified as moderately mobile, while SDS-46851 would be classified moderate to mobile.

61

5.9

MOBILITY IN SOIL

Summaries of soil column leaching studies were submitted. The general conclusions from these studies confirmed the mobility classifications previously given. The studies showed that whilst chlorothalonil had a low potential to leach in the various soil types tested, the mobility of the soil metabolites would greatly depend on their respective mobility, amount formed and their rate of degradation. A summary of the respective leaching potential of the chlorothalonil and its soil metabolites were given as; (High mobility) SDS-46851 > SDS-47525 > SDS-3701 > SDS-47523/4 = SDS-19221 > chlorothalonil (immobile) (Unpublished, 1995b).

5.10
5.10.1

LEACHING FROM A TREATED SURFACE

LABORATORY METHOD

In 1990, a study was carried out to determine the leach rate of chlorothalonil from a surface painted with a water based preservative formulation (6.97 % chlorothalonil). The study was performed in accordance with the Interim Draft ASTM Standard Test Method for Organotin Release Rates of Antifouling Coating Systems in Sea Water dated 9-27-88. Modifications were made for the analytical determination to enable the analysis of chlorothalonil concentrations in seawater. The study was also conducted to GLP. Three 6.35 cm diameter polycarbonate test cylinders were coated with the formulated paint to provide an exposure area of 200 cm2 with a paint film thickness of at least 0.01 cm. The coated cylinders were then aged in air for 7 1 d at 25 oC prior to the leach rate determinations. The cylinders were then placed in a holding tank containing synthetic seawater which was monitored to maintain temperature (25 C), pH (7.8 - 8.2) and salinity (not stated) content. A positive low velocity flow (5 l min-1) within this tank was established by an external pump loop which circulated the water through an activated carbon filter to maintain background seawater copper levels below 100 g l-1, as recommended by the guideline. On days 1, 3, 7, 10, 14, 21, 24, 28, 31, 35, 38, 42 and 45 the cylinders were transferred to test polycarbonate beakers and leached by rotation at 60 5 rpm for 1 h in synthetic seawater maintained at 25 C. Between sample periods, the cylinders were placed back in the holding tank. Samples taken during the leaching phase were subjected to solid phase extraction, eluted by solvent and analysed for chlorothalonil by HPLC using a variable wavelength UV detector. The separation technique was validated with the use of standard solutions in seawater and the recovery was demonstrated to be > 95 %. The levels of chlorothalonil leached from the painted cylinders were initially low and erratic, but stabilised at a higher level for the remainder of the test period. HPLC data from samples taken during the first two weeks indicated decomposition products were present, although these were not investigated further. Chlorothalonil release rates were calculated in micrograms per square centimetre of paint sample per day, and the cumulative rate in micrograms or milligrams per day throughout the six week test period. The mean release rate of 3.06 g cm-2 d-1 was calculated by averaging individual release rate measurements taken 62

from day 21 through to the last day of sampling (day 45) , however, a cumulative rate was not reported (Unpublished, 1990). In 1995, a study was conducted to determine the rates at which chlorothalonil (2.8 % w/w) and copper (cuprous thiocyanate 12.2 % w/w) were released from a yacht antifouling coating into synthetic seawater maintained at 25 2 oC. The study followed the ASTM D5108-90 guideline (Standard Test Method for Organotin Release Rates of Antifouling Coating Systems in Seawater) and was conducted to GLP. Three test cylinders were prepared using 8-oz glass bottles masked to expose an area of 8 cm x 19 cm or 152 cm2. Over a period of 17 d, five coats of the test antifouling paint were applied to each test cylinder for a sample thickness of 229 m. Each coat was allowed to dry completely before adding the next coat. An uncoated test cylinder was used as a blank. The final coat of paint was allowed to age for seven days in a 25 C environmental chamber. The coated cylinders were placed in a holding tank of synthetic seawater where the concentrations of chlorothalonil and copper were kept low by fortnightly seawater renewal, and by circulating the seawater through a carbon filter. The pH of the holding tank was monitored daily and adjusted to between 7.8 and 8.2. The temperature of the holding tank was monitored daily and maintained between 23 and 27 C. Weekly samples were taken from the tank for copper and chlorothalonil analysis. The test cylinders were removed from the holding tank on days 1, 3, 7, 10, 14, 21, 24, 28, 31, 35, 38, 42 and 45, and transferred to baffled measuring vessels containing 1500 ml of fresh seawater maintained at 25 1 C. The cylinders were then rotated at 60 5 rpm for 1 h. After the rotation period was completed, the cylinders were immediately removed from each measuring vessel and returned to the holding tank. Two sub-samples were removed from each vessel, one sub-sample was analysed for chlorothalonil and one for copper. Some of the samples were frozen prior to analysis (storage was limited to less than one week). Chlorothalonil samples were analysed in triplicate by GC (96.1 % recovery) and copper analysis was carried out by Inductively Coupled Plasma-Optical Emission Spectroscopy (ICP-OES). The 14-day cumulative chlorothalonil release was 51 g cm-2, from which an average daily release rate of 3.18 g cm-2 d-1 was calculated. For comparison, the 14-day cumulative copper release was 510 g cm-2, equating to a daily average of 27.38 g cm-2 d-1 (Unpublished, 1995a). 5.10.2 CALCULATION METHOD In 1998, a brief document outlining a leaching rate calculation for two formulations of antifouling paint was submitted in support of the chlorothalonil review. The study used a calculation method agreed amongst antifouling paint manufacturers as suitable for all antifouling components with the exception of TBTO and TBT - methacrylate copolymers. This method calculates the amount of ai applied to a surface, and assumes 100 % of the ai leaches from the paint over the in-service lifetime of the paint. A 14-day cumulative leaching of chlorothalonil from paint type I (3 % w/w) and II (5 % w/w) were reported as 29.80 and 51.06 g cm-2 respectively. The daily average leaching rates were 63

also calculated and reported as 0.99 g cm-2 d-1 for paint type I, and 1.70 g cm-2 d-1 for paint type II. However, no validation data has been supplied with this study (Unpublished, 1998).

5.11
5.11.1

BIOCONCENTRATION
BLUEGILL SUNFISH

In 1972, a study was conducted to determine the bioconcentration, distribution and elimination of 14C-radiolabelled chlorothalonil (purity unknown) in bluegill sunfish (Lepomis macrochirus) under flow-through conditions. The study was not carried out to a recognised guideline. A proportional diluter system was set up to deliver test solution to two replicate 30 l aquaria, at a flow rate of 6 l h-1. One hundred bluegill sunfish, with body lengths of 5 - 7.5 cm and mean body weights of 3.0 g, were placed in each aquarium. Aerated well-water, which was maintained at 18 0.5 C (total hardness of 40 mg l-1 CaCO3, a dissolved oxygen (DO) level > 5 mg l-1 and a pH of 7.1) was delivered to both aquaria for three d before exposure to chlorothalonil was initiated. A radiolabelled 14C-chlorothalonil stock was prepared in acetone and delivered to one test aquarium only at a nominal test concentration of 0.01 mg l-1 chlorothalonil. The remaining aquarium was maintained as a positive control (no further details given). The fish were exposed to the test solutions for a total of 28 d, following which time the remaining fish from both aquaria were transferred to uncontaminated well-water for a further 14 d. It was not clear in the report whether flow-through conditions were maintained for the depuration phase. The fish were fed ad libitum on a dry pellet diet throughout the study. No details of water quality measurements, or the photoperiod used during this study were reported. Five fish were sampled from each aquarium prior to the beginning of the test and then after 1, 3, 7, 10, 14, 21 and 28 d of exposure. Once sampled, each fish was eviscerated, freeze dried and the edible tissue (only) radiometrically assayed by combustion analysis. The distribution of chlorothalonil residues between edible and non-edible tissues was only investigated after 28 d of exposure, by additional radiometric combustion analysis of the visceral mass from each fish sampled. In addition the relative amounts of non-polar and polar residues were investigated by assaying solvent extracts (hexane and methanol respectively) of pooled edible portions from five fish exposed to chlorothalonil for 28 d. After being transferred to clean well-water a further five fish (chlorothalonil exposed fish only) were sampled on days 1, 3, 7, 10 and 14. Water samples (500 ml) were also taken at each sampling point, extracted by methyl chloride and radiometrically assayed using a scintillation spectrometer. Recovery rates of 94 % and a detection limit of 0.001 mg l-1 were reported for the analytical techniques used. However, the results all assumed that measured radioactivity was proportional to chlorothalonil concentration, metabolites were not considered. From the water samples, a mean measured concentration over the period of exposure was reported as 0.003 mg ai l-1, which equated to 30 % of the nominal. The report author suggested that this was due to the adsorption of the test material to the surface of the test system since the levels fell rapidly within the first 24 h to 30 % of the nominal concentration, and remained constant thereafter.

64

Statistical analysis, (t-test for unpaired observations with unequal variances) showed that there were no significant differences (p 0.05) between the residues found in the fish sampled after 3, 7, 10 14, 21 and 28 d exposure. This indicated an equilibrium had been reached after 3 d of exposure, and this concentration was maintained (mean 0.56 0.09 mg ai kg-1). The reported bioconcentration factor (BCF) for edible fish tissues exposed to chlorothalonil was 60, however, this was calculated using the nominal test concentration of 0.01 mg ai l-1. HSE, using the mean measured exposure concentration of 0.003 mg ai l-1, have recalculated the BCF to be 187. The non-edible tissue analysed after 28 d exposure to chlorothalonil demonstrated far greater bioconcentration than the edible tissues at 8.6 2.7 mg ai kg-1, which equated to a reported BCF of 860 based on the nominal concentration. Again, HSE recalculated the BCF for non-edible tissue to have been 2867, based on the mean measured concentration. Solvent extraction of the edible tissue resulted in 16 and 18 % of the 14C-residues being identified as polar and non-polar extracts respectively, indicating that degradation of the parent compound had occurred. This was not investigated or discussed further by the study author. Approximately 50 % of the edible tissue residue level was lost after 10 d exposure to uncontaminated well-water. The author predicted that, if the rate of elimination remained linear, it would take 56 d for the residue levels in the edible tissue to return to that of the exposure concentration. However, it was not clear whether the author used the nominal or mean measured test concentration to calculate this. No data was presented or discussed with regards to the depuration of chlorothalonil from the non edible fish portion (Unpublished, 1972). In 1980, a study was conducted to evaluate the bioconcentration, distribution and depuration of 14C-chlorothalonil residues in bluegill sunfish (Lepomis macrochirus) exposed to 14Cchlorothalonil (99.9 % purity) under continuous flow-through conditions. The study was not carried out to recognised guidelines or to GLP. However, it was carried out to an in-house laboratory protocol, which was submitted with the report and included a quality assurance certificate. The test system consisted of four 100 l glass aquaria, two of which served as controls and two of which served as test aquaria. Each aquarium contained 70 l of aerated well-water. A proportional diluter system was used for the intermittent introduction of the test solution into the duplicate test aquaria, and well-water into the duplicate control aquaria at a flow rate of 500 ml min-1 per aquaria. The control aquaria received an equal amount of solvent carrier (ethanol) as that used in preparation of the nominal test solution. Each aquarium was immersed in a circulating water bath maintained at around 22 C. A nominal test concentration of 0.008 mg 14C-chlorothalonil l-1 (1/10th previously established in-house NOEC for bluegill sunfish) was selected, and the test system calibrated to this concentration 24 h prior to the test initiation. Bluegill sunfish, of mean length 69 mm and weight 4.9 g, were acclimated to the laboratory well-water (total hardness 260 mg CaCO3 l-1, DO 9.3 mg l-1 and pH 7.7) 14 d prior to the test initiation. Fish were acclimated to the test temperature of 22 C, 72 h prior to the test

65

initiation, and were fed ad libitum throughout the acclimation and subsequent test period. At the start of the test 120 fish were placed in each of the four aquaria and sampled on days 1, 3, 7, 10, 14, 22 and 30. At the end of the exposure period, the input of 14C-chlorothalonil to the test system ceased and all but three inches of the test water siphoned off from each aquarium and replaced with 70 l clean, uncontaminated well-water. This procedure was repeated one further time. The fish were then exposed to uncontaminated flowing well-water for 14 d, and sampled on days 1, 3, 7, 10 and 14. Water samples, taken at each sampling point during both the exposure and depuration phases, were extracted and analysed by LSC. Water quality parameters were measured throughout the study. The sampled fish were analysed either whole or divided into edible and non-edible tissue. The fish or pooled tissues were homogenised prior to total combustion analysis or LSC (whole fish only). Additional fish and water samples from each test aquaria were sampled during the exposure and depuration phases to provide sufficient material which was frozen and stored until subsequent characterisation analysis was carried out. The water quality parameters remained acceptable throughout the study period, except for the dissolved oxygen levels which fell to 2.4 mg l-1 on day 3 of the exposure phase. This was as a result of a malfunction of the diluter apparatus. However, this fault was rectified immediately and no adverse effects were seen in the fish or bioconcentration results. The mean measured concentration of 14C-chlorothalonil during the exposure phase was 0.0072 mg l-1 (90 % of nominal) and during the depuration phase the maximum concentration recorded was 0.0011 mg l-1. The bioconcentration profile of 14C-residues demonstrated that a plateau of 14C-residues was attained by day 14 of the exposure phase, after which the residues were seen to decline. By day 30, BCFs for whole fish, fillets and viscera were reported as 264, 76 and 514 respectively. At the end of the 14 d (day 44 of test) depuration phase 80, 46 and 85 % of the accumulated 14C-residues had been eliminated from the whole fish, fillet and viscera samples respectively (Unpublished, 1980b). The characterisation and quantification of 14C-residues in the water and bluegill sunfish tissues sampled and stored frozen on days 0, 7, 10, 14, 22 and 30 of the previous exposure phase, and on days 3, 7, 10 and 14 of the previous depuration phase. Tissue extracts from either whole fish, fillet or viscera were prepared using an appropriate solvent. Unextracted 14 C-residues were quantified using biological oxidation techniques (no further details were given). The 14C-residues in both the water samples and fish tissue extracts were selectively partitioned into an organic solvent separating the 14C-residues into organic and aqueous phases. Characterisation of the various fractions were carried out using TLC and quantified by LSC. The water samples, subjected to selective partitioning, showed that the majority of the 14Cresidues remained in the aqueous phase. The concentration of chlorothalonil decreased from 0.0024 mg l-1 on day 0, to non detectable levels by day 22 of the exposure phase. Table 5.14. demonstrates that 14C-chlorothalonil was largely degraded into 14C-DS-3701 and 14C-19221, in addition to which were some unidentified polar residues (not discussed further).

66

Table 5.14: Characterisation and Quantification of 14C-residues in Water Sampled from a Flow-through System
14

C-chlorothalonil Equivalents mg l-1

14

DAY 0E 7E 14E 22E 30E

E 2

Total 14Cresidues 0.0052 0.0061 0.0059 0.0063 0.0069

14

C-chlorothalonil 0.0024 0.0014 0.0004 ND5 ND5

C-DS-3701 ---4 0.0005 0.0009 0.0014 0.0011

14

C-DS-19221 ---4 0.0010 ND5 ND5 0.0003

- exposure phase. - origin of chromatogram. 3 - corresponds to Rf region between that of DS-3701 and the origin. 4 - no assay conducted. 5 - 14C-radioactivity in sample was < twice that of the background level for chlorothalonil.

14

C-

In the whole fish, fillet and viscera, 35 - 69 % of the total 14C-residues were non-extractable, and less than 6 % of the total 14C-residues could be identified (see Table 5.15.).

Table 5.15: Characterisation and Quantification of 14C-residues in Bluegill Sunfish Following 14C-chlorothalonil Exposure Under Flow-through Conditions
14

C-chlorothalonil Equivalents mg l-1 unextracted 14Cresidues 30E 14D 1.25 0.35 2.58 0.27 0.16 0.32 Cchlorothalonil 30E 14D 0.04 0.02 0.05 ND ND ND
14

Day Whole fish Fillets Viscera

E

total 14Cresidues 14D 30E 2.29 0.63 4.79 0.40 0.29 0.61

- exposure phase. - depuration phase. 2 - origin of chromatogram. 3 - corresponds to Rf region between that of DS-3701 and the origin. 4 14 - C-radioactivity in sample was < twice that of the background level for 14C-chlorothalonil.
D

Under the conditions of this study 14C-chlorothalonil in water degraded to 14C-DS-3701, 14CDS-19221 and polar water-soluble 14C-residues. Although chlorothalonil was accumulated in fish tissues, the majority (35 - 69 %) of the quantified 14C-residues were characterised as being unextractable. A further 28 - 47 % were characterised to be polar 'water soluble' residues. Tissue analysis following the 14 d depuration detected no chlorothalonil. However, 67

no stability data was presented on the effects of storing the frozen fish and water samples, therefore, the results should be treated with caution (Unpublished, 1981c). 5.11.2 CHANNEL CATFISH

In 1981, a study was carried out to determine the bioconcentration and distribution of 14Cresidues in channel catfish (Ictalurus punctatus) exposed, under static conditions, to aged-soil spiked with 14C-chlorothalonil (99.7 % purity). The study also investigated the depuration of the 14C-residues under flow-through conditions. The study was not carried out to recognised guidelines or to GLP. However, it was carried out to an in-house laboratory protocol, which was submitted in full with a quality assurance statement. Four samples of 74.5 kg of sieved sandy loam soil (73 % sand, 23% silt and 4% clay) were prepared. Two samples were fortified with 14C-chlorothalonil spiked soil at a nominal concentration of 10 mg kg-1 soil. The remaining two samples were kept as controls. Each of the spiked and un-spiked soil samples were then transferred to stainless steel test tanks (to which chlorothalonil had previously demonstrated negligible adsorption). The soil samples were then aged for 30 d under conditions of ambient temperature (22 2 C), 16L:8D and 15 - 20 % soil moisture. Following this, 1000 l of well-water was added to the tanks and this was left to equilibrate for 3 d. The tanks were constantly aerated throughout the equilibration and subsequent exposure periods. After 3 d, 255 catfish were added to each tank and exposed for 26 d. After this time all the remaining catfish were transferred to aquaria containing uncontaminated flowing water for a 14 d depuration period. Soil samples (100 g) were taken from each tank on day 0 and 30 of the ageing phase, then on days 0, 1, 3, 7, 10, 14, 22 and 26 of the exposure phase. Each time the soil sample was filtered, air dried, ground and analysed by combustion radioanalysis. Five fish were sampled from each tank on days 1, 3, 7, 10, 14, 22 and 26 of the exposure period, then on days 1, 3, 7, 10 and 14 of the depuration period. Two of the fish were retained for whole fish samples, the remaining three were dissected and pooled as edible or non-edible tissue. Each pooled portion, including whole fish tissue, was then mixed with dry ice, homogenised, and subsamples taken for combustion radioanalysis. Water samples (500 ml), taken from each tank on all sampling days, during exposure and depuration were analysed by LSC. The remaining water was frozen and retained for metabolite characterisation (no analytical method described). The results of the soil radioassay indicated that the amount of 14C-residue (assumed to be 14Cchlorothalonil) decreased from 7.7 mg ai kg-1 soil on day 0 of the ageing period to 6.9 mg ai kg-1 soil on day 26 of the exposure period. This represented a 10 % reduction in 14Cradioactivity in the soil. Radioassay results of the water samples indicated that the exposure level of 14C-residues increased from a mean of 0.069 mg ai l-1 on day 0 of the exposure phase to a mean of 0.34 mg ai l-1 on day 26. Analysis of 14C-residues (assumed to be 14C-chlorothalonil) in the edible, non-edible and whole fish tissues suggested that after a peak on day 3, a plateau was reached by day 7 of the exposure phase with 0.42, 0.93 and 0.74 mg ai kg-1 tissue respectively. The maximum and final mean 14C-tissue concentrations plus corresponding BCFs for edible tissue, non-edible tissue and whole fish are presented in Table 5.16.

68

Table 5.16: Summary of Chlorothalonil Tissue Concentrations and Bioconcentration Factors Max. concentration mg ai kg-1 tissue (day) 0.83 (1) 2.4 (3) 1.9 (3) Max. BCF 9.4 20 16 Tissue concentration (day 26) 0.33 1.4 0.85 BCF (day 26) 0.97 4.1 2.8

Edible Non-edible Whole fish

Once transferred to untreated water, the 14C-radioactivity decreased to 0.061, 0.079 and 0.079 mg ai kg-1 in the edible, non-edible and whole fish tissues respectively by day 14 of the depuration period. This represented 82, 94 and 92 % losses respectively. The results of this study demonstrated that chlorothalonil did not significantly bioconcentrate in tissues of the channel catfish under the conditions of the study. The exposure period of this study was prematurely terminated at 26 rather than 30 d following several recorded deaths in the treated fish. After 26 d it was also noted that the surviving fish were becoming increasingly stressed and surfacing. However, after only 3 d exposure to clean, flowing water, the affected fish appeared to have recovered (Unpublished, 1981d). On each of the sampling occasions previously mentioned portions of water, soil, whole fish, edible and non-edible fish tissues were taken and frozen. However, no stability data was presented on the effects of storing the frozen fish and water samples, therefore, as for the previous characterisation study these results should be treated with caution For the soil and fish tissues samples, the 14C-residues were prepared and analysed as detailed in the previous characterisation study. Unextracted 14C-residues were quantified using biological oxidation techniques (no further details were given). Under the conditions of this study, 14C-chlorothalonil degraded in soil with a reported halflife of 30 d. After the 30 d ageing period no loss of total 14C-radioactivity was reported, indicating that neither 14C-chlorothalonil or its degradation products volatilised from the test system. During the exposure phase of the study the concentration of 14C-chlorothalonil in soil decreased steadily with time from 4.06 to 1.06 mg kg-1 (73.9 % loss), by day 26. In water, the total concentrations of 14C-residues increased from 0.071 mg l-1 on day 0 to 0.397 mg l-1 on day 26. 14C-chlorothalonil concentrations in water accounted for less than 3 % of the total 14 C-residues, whereas, 52 - 72 % was characterised as 14C-DS-3701. Minor (< 10 %) 14Cresidues in water were reported which were characterised to be either more polar or less polar than DS-3701. During the exposure phase, fish tissue analysis revealed 14C-chlorothalonil and 14C-DS-3701 were both major 14C-residues (see Figure 5.2), with the majority of the accumulated 14Cresidues bioaccumulated in the non-edible tissue portions (Unpublished, 1981e).

69

1.2 1 0.8 0.6 0.4 0.2 0 7

unextractable chlorothalonil

DS3701 unidentified II

unidentified I unidentified III

14

22

26

29

40

2.5 2 1.5 1 0.5 0 7 14 22 26 29 40

Mean 14C-residues (mg/kg tissue)

1.5

0.5

0 7 14 22 26 29 40

test duration

Figure 5.2: Characterisation and quantification of major (>10%) 14C-residues in edible (top), non-edible (middle) and whole fish (bottom) tissues sampled through out the 26 d exposure period, and 14 d depuration period

70

5.12 SUMMARY OF FATE AND BEHAVIOUR

Chlorothalonil was shown to be hydrolytically stable at pHs 7 and 5 (22 C) under sterile conditions over a period of 49 d. However, at pH 9 a half-life of 38 d was calculated and the metabolites DS-19221 (54 %) and DAC-3701 (22 %) identified. Further investigations also revealed that DAC-3701 was extremely resistant to hydrolysis at all pHs tested. Soil spiked with chlorothalonil and SDS-3701 was shown to be photolytically stable. Chlorothalonil was shown to undergo aqueous photolysis and a half-life of 64.7 d was calculated (exposure equivalent to 30, twelve-hour sunlight days). However, it is unlikely that chlorothalonil would be significantly removed from the aquatic environment via this route. No study addressing the effects of chlorothalonil on microbial activity in sewage sludge was submitted. However, three US soil studies were presented which demonstrated that chlorothalonil (2.5 and 25 mg ai kg-1 soil) did not adversely affect several key microbial processes. Biodegradation in soil and water-sediment systems were investigated, and a full degradation pathway suggested (see Figure 5.3.), however, full studies were not submitted. It was predicted that the degradation processes of chlorothalonil in soil was the same regardless of aerobic or anaerobic conditions. In both cases, the major 14C-residues were identified as either bound, SDS-3701 or SDS-19221. Degradation half-lives in soil were given as between eight and 36 d for chlorothalonil and between six and 43 d for the major soil metabolite SDS-3701. In aerobic sediment-water systems (fresh and seawater), chlorothalonil underwent rapid degradation with half-lives of < 2 h suggested. The metabolites identified as a result of aquatic metabolism, in sediment-water systems, were not the same as those identified following chlorothalonil degradation in soil. Under aerobic aquatic conditions two major metabolites (SDS-67042 and SDS-67042 sulphoxide) and three minor metabolites were reported (SDS-66432, SDS-13353 and SDS-3701). It should be noted therefore that the metabolites of concern in the aquatic environment have not been previously addressed due to the previous reviews of chlorothalonil concentrating solely on the agricultural use of chlorothalonil, where the contamination of the aquatic environment was considered unlikely. Literature studies (Davies, 1988; Gajbhiye et al., 1989 and Walker et al., 1988), provided additional evidence that in sediment-water systems chlorothalonil was primarily removed from the water column via adsorption to sediments where subsequent degradation processes occurred. These studies suggested that chlorothalonil was primarily removed from the water phase as a result of adsorption, where subsequent degradation processes occurred. The degradation rate in sediment was demonstrated by Davies (1988) to be dependant on temperature (10 C rise, gave rise to 50 % reduction in T1/2), which along with evidence from Walker et al., (1988) using sterile (T1/2 5.03 d) and non-sterile (T1/2 1.79 d) sediment water systems reinforced the importance of biodegradation. Callow and Willingham (1996) investigated the loss of chlorothalonils efficacy (toxicity) over an eight week period using a seawater bioassay. They reported half-lives in biocidal activity of > 8 weeks in natural seawater, or ~ 8 weeks in seawater with enhanced biomass, which were both greater than those previously described in the literature. However, the results of this study were based on toxic activity in water alone (no sediment) and could not distinguish degradation products. A US soil study reported that chlorothalonil was readily bound to soils (57.1 - 93.7 %) and did not desorb (1.9 - 24.2 %) to any great extent. However, because of the methodology,

71

HSEs Technical Secretariat did not consider that the desorption potential of chlorothalonil was adequately investigated, and potentially underestimated the likely remobilisation of chlorothalonil from aquatic sediments. Summary data from the EU review of chlorothalonil further supported the adsorption study reporting chlorothalonil to be of low mobility in various soil type. It was also reported that the soil metabolites were more mobile that chlorothalonil. Two laboratory and one calculation method was submitted to address the leaching potential of chlorothalonil from antifouling paints. The laboratory studies both followed the draft ASTM guideline, and reported comparable mean daily leaching rates of 3.06 and 3.18 g cm-2 d-1 from two antifouling paint formulations containing different levels of chlorothalonil. The theoretical study assumed that all the active ingredient contained in a formulation would have leached out over the expected lifetime of the paint. The reported mean daily leaching rates of 0.99 and 1.70 g cm-2 d-1, were notably less than those gleaned from the laboratory studies. Five studies investigated the bioaccumulation of chlorothalonil in two different fish species, bluegill sunfish and channel catfish. The BCFs reported were higher for the bluegill sunfish (up to 2867 in non-edible portion) than for those measured in the channel catfish (maximum of 20 in non-edible portion). However, the reasons for these differences are probably reflected in the experimental design with the bluegill sunfish being exposed directly to chlorothalonil and the catfish exposed to soil leachate. Both species were shown to reach a bioaccumulation equilibrium within 14 d and depurated at least 46 % of the accumulated ai in the same period of time under the selected test conditions. Further to this, the BCF values reported in each study consistently demonstrated preferential sequestration of chlorothalonil and/or its metabolites by the non-edible tissue portion. This maybe due to the presence of a detoxification process occurring in such organs as the gills, liver, spleen and kidneys. It should be noted that the BCFs recorded for both fish species studied were not calculated for chlorothalonil but for 14C-residues, and that subsequent characterisation studies revealed that chlorothalonil was not the major residue identified in water. The major 14C-activity in the bluegill sunfish tissues was characterised as unextracted, whereas the 14C-residue in the channel catfish was largely 14C-chlorothalonil.

72

CN Cl Cl Cl

CN Cl

O Cl S NH Cl CN Cl Cl OH CN

SDS-67042
CN Cl

SDS-3701
Cl Cl OH O Cl CN

CONH2 Cl

SDS-3701 Photolysis

Cl Cl Cl CN S O NH CN Cl Cl

SDS-67042 Sulfoxide

SDS-19221 Aerobic aquatic Fresh or Saltwater

Cl Cl CN

[Bound Residue]

Aerobic or Anaerobic Soil

[Bound Residue]

+ +

+
CN Cl Cl

CONH2 Cl

SDS-2787 Hydrolysis pH 9.0

Cl Cl

CN

Cl OH

CN Cl

CONH2 Cl

SDS-47523/4

SDS-3701

+
CN Cl Cl Cl Cl CN Cl

CN Cl

SDS-19221

CN Cl

Cl OH

CONH2

Cl SH

CN Cl

SDS-47525

SDS-13353

+
CN Cl Cl

Cl OH

CN

+
COOH Cl

SDS-3701
Cl Cl GLUT CN Cl CONH2

SDS-46851

SDS-66382

+
CN TULG Cl

Cl GLUT

CN

SDS-66432

Figure 5.3:

Metabolism of chlorothalonil in the environment (Unpublished, 1995b)

73

6 ECOTOXICOLOGY
The use of chlorothalonil as an antifoulant on vessels and in aquaculture was first reviewed by committees in 1996. [Aquaculture uses have now been voluntarily withdrawn] The following environmental toxicological data requirements were set; a) A reasoned case demonstrating that chlorothalonil can be used without adverse effects to the environment.

And full test reports for:f) g) Acute toxicity to a suitable juvenile fish species (96 hr flow-through), and Acute toxicity (48 hr EC50) to a single pelagic non-target invertebrate species (e.g. Daphnia magna), both of which were submitted in summary form for the review in 1996.

With the exception of a), the required studies have been submitted in full. The company also submitted summary data, which had been prepared in support of the review of chlorothalonil as an agricultural fungicide under the European Plant Protection Products Directive (PPPD) (Unpublished, 1995b). Unfortunately, because of the summarised format, it was not possible to evaluate these data in the absence of full study reports. Therefore, only brief details of the additional studies, where endpoints are considered pertinent to the use of chlorothalonil as an antifoulant, have been included in this section. In addition to the studies provided by the company, a number of published studies have been reviewed and included with the relevant sections.

6.1

TOXICITY TO FRESHWATER ALGAE

A summary of a 5 d exposure study investigating the toxicity of technical chlorothalonil upon Selenastrum capricornutum was submitted. Triplicate flasks containing algal suspensions plus nominal concentrations of 0.025, 0.05, 0.1, 0.2 and 0.4 mg ai l-1 (chlorothalonil, solvent carrier, dimethyl formamide - DMF) were maintained at 24 C in an orbital incubator for 5 d, in addition to three solvent control flasks. The algal growth rate was determined by measuring the number of algal cells ml-1 after 72, 96 and 120 h of exposure using a Coulter Counter. After 72 h, the number of cells at 0.1 mg ai l-1 was significantly higher than that reported for the control samples, however, this was not significant by day 5. A stimulation of the algal growth rate was also noted at 0.025 mg ai l-1 on day 5. The 0.2 and 0.4 mg ai l-1 exposure levels resulted in significantly reduced algal growth compared to the controls by day 5. The 5 d EC50, LOEC and NOEC was reported as 0.21, 0.2 and 0.1 mg ai l-1 respectively (Unpublished, 1995b).

74

6.2 . ACUTE TOXICITY TO INVERTEBRATES

6.2.1 FRESHWATER INVERTEBRATES

In 1977, a 48 h acute toxicity study was carried out to determine the toxicity of technical grade chlorothalonil (purity unreported) to Daphnia magna under static conditions. The study was reported to be based on protocols in "Methods for acute toxicity tests with fish, macro-invertebrates and amphibians" (EPA 1975), but was not carried out to GLP. Nominal test concentrations of 6.8, 19, 53, 150 and 410 g ai l-1 were prepared by dissolving the required amount of the test compound in acetone which was then added to the diluent; reconstituted well-water (hardness 60 mg l-1 as CaCO3, pH 7.2). Three triplicate beakers, each containing five daphnids were set up with 150 ml of each test concentration, along with a solvent and dilution water-only control. The dissolved oxygen concentration, pH and temperature of the test solutions were measured at both the initiation and at the termination of the test in the highest, lowest and middle test concentrations. Observations for immobility were made and recorded after both 24 and 48 h. The temperature was maintained at 22 1C throughout the test. Dissolved oxygen concentrations ranged from 7.9 to 8.8 mg l-1 (90 - 100 % saturation), and pH ranged from 7.2 to 7.4. No mortalities or sub-lethal effects were observed in either control group. The nominal 24 and 48 h EC50 values (with 95 % confidence limits) calculated using least squares regression analysis, are presented in Table 6.1. Table 6.1: Acute Toxicity of Chlorothalonil to D. magna EC50 g ai l-1 (95% C.L) 120 (48.4 - 299) 70 (34.2 - 143)

24 h 48 h

A NOEC, at which no effects were observed, of 6.8 g ai l-1 was reported (Unpublished, 1977). In 1992, a 48 h acute toxicity study was carried out to determine the toxicity of a chlorothalonil formulation (54 % ai) to Daphnia magna under flow-through conditions. The study protocol followed FIFRA guideline 72-2, and was carried out to GLP. The flow-through test conditions were achieved using an intermittent-flow proportional diluter which was in operation for two days prior to the test to allow the equilibration of the diluter apparatus and exposure vessels. Each test vessel (1.8 l) received approximately six volume replacements per day. All vessels were maintained in a water bath at 20 1C with a 16L:8D photoperiod. Twenty neonates (ten per replicate) were exposed to mean measured concentrations of 226.8, 140.4, 86.4, 49.14 and 27 g chlorothalonil l-1. Two further replicates of ten neonates were exposed to a dilution water-only control. The dilution water, fortified well-water, had a reported total hardness of 170 mg CaCO3 l-1, and a pH range of 8.1

75

- 8.3. Observations of immobility, sub-lethal effects and physical characteristics of the test solutions were recorded after 24 and 48 h exposure. Water quality was measured daily in all treatment vessels throughout the exposure period. All test vessels were sampled after 24 and 48 h exposure and analysed for chlorothalonil by GC using ECD (recovery 101 6 %). No undissolved test material was observed in the diluter system or exposure solutions during the study. Water quality parameters were unaffected by the concentrations of chlorothalonil formulation tested with reported ranges of pH at 8.2 - 8.3, 93 - 105 % O2 saturation and 170 180 mg CaCO3 l-1 total hardness. After 48 h exposure 100, 95 and 30 % immobilisation was observed in daphnids exposed to mean measured test concentrations of 226.8, 140.4 and 86.4 g chlorothalonil l-1 respectively. EC50 concentrations, calculated by probit analysis, are presented in Table 6.2. Table 6.2: Acute Toxicity of Chlorothalonil to D. magna EC50 g ai l (95% C.L) 183.6 (151.2-232.2)
-1

24 h 48 h

97.2 (86.4-108.0)

Sub-lethal effects (e.g. erratic swimming) were observed in many of the mobile daphnids in the groups exposed to 140.4 and 86.4 g chlorothalonil l-1. No effects were observed in the control daphnids or those exposed to the two lowest mean measured test concentrations, 49.14 and 27 g ai l-1. Therefore a NOEC of 49.14 g ai l-1 was established (Unpublished, 1992a). 6.2.2 ESTUARINE/MARINE INVERTEBRATES

An acute 96 h toxicity test to determine the toxicity of technical chlorothalonil to penaeid (pink) shrimp (Penaeus duorarum) was carried out in natural filtered seawater under static conditions with no aeration. This study was submitted in summary form only. Six replicate vessels each containing 2 shrimp were prepared at nominal concentrations of 75, 100, 150, 200 and 300 g ai l-1. Solvent (acetone) and a dilution water-only controls were run concurrently. Mortality, general condition and behaviour was assessed after 6 h exposure then thereafter every 24 h for a 96 h period. Mortalities and signs of toxicity were only observed at 100 g ai l-1 and above. The cumulative % mortality in the 100, 150, 200 and 300 g ai l-1 exposure groups was 67, 42, 33 and 83 % respectively. The 96 h LC50 was reported as 165 g ai l-1 (95 % C.L. = 100 - 270 g ai l-1), and the NOEC reported as 75 g ai l-1 (Unpublished, 1995b).

76

A second study, submitted only in summary form, investigated the acute toxicity of technical chlorothalonil upon the shell deposition in Eastern Oyster (Crassostrea virginica). The study was conducted in natural filtered seawater under flow-through conditions with no aeration. Single replicates containing ten oysters were exposed to six nominal chlorothalonil concentrations (1, 2, 4, 8, 16 and 32 g ai l-1) prepared with acetone as a carrier. A dilution water-only and a solvent control was run concurrently. Observations of the oysters general condition were made daily, and the percentage of new shell growth was measured and compared between the treatment groups at the end of the 96 h exposure period. Analysis of the test solutions reported 80 - 96 % of the nominals were achieved with the exception of the 1 g ai l-1 level which achieved only 60 %. New shell growth was significantly reduced at concentrations of 8 g ai l-1 and higher. The 96 h EC50 value for shell deposition (the concentration of test material estimated to reduce new shell growth by 50 % compared to the solvent control) was calculated as 7.3 g ai l-1, and a NOEC of 4 g ai l-1 was reported (Unpublished, 1995b).

6.3
6.3.1

CHRONIC TOXICITY TO INVERTEBRATES

FRESHWATER INVERTEBRATES

A 42 d (21 d per generation) chronic toxicity study was carried out to determine the effect of technical chlorothalonil on Daphnia magna under flow-through conditions without aeration. The study was submitted in summary form. Quadruple vessels, each containing 20 neonates (< 24 h old), were prepared at each mean measured concentration of 3.8, 6.5, 16, 35 and 79 g ai l-1. No details of controls were given. The first generation were exposed for 21 d, during which daily observations for mortality and offspring were made. On day 20/21 the offspring from each test concentration were used to initiate a second generation. [Since only one daphnid survived the highest test concentration, control offspring were used to initiate the second generation for this exposure level]. The second generation were exposed for a second period of 21 d, during which conditions and observations were as for the first generation. Survival of the first generation females were comparable to the controls with the exception of the highest concentration, 79 g ai l-1, in which only one female survived. No effects on offspring production were noted during the exposure period of the first generation at concentrations of 35 g ai l-1 and below. At 79 g ai l-1, the total number of offspring produced per female was significantly reduced when compared to the control. During the second generation exposure period the survival of the daphnids exposed to the highest test concentration of 79 g ai l-1 were again significantly reduced, with only 5 % survival by day 42. Adult survival was not affected at exposure concentrations of 35 g ai l-1 and below. Offspring production (number per female) was significantly higher at the highest concentration tested (79 g ai l-1), despite adult survival being poor.

77

Since adverse effects on adult survival and offspring production was recorded at 79 g ai l-1 for two consecutive generations the MATC and NOEC for Daphnia magna chronically exposed to technical chlorothalonil was 35 g ai l-1 (Unpublished, 1995b). HSE considered that the results confirmed that adverse effects on adult survival and offspring production would occur in daphnids exposed to 79 g ai l-1. However, since the daphnids used in the second generation test were obtained from control adults they did not fulfil second generation, and the risk to D. magna from chronic exposure to chlorothalonil was not adequately addressed. 6.3.2 ESTUARINE/MARINE INVERTEBRATES

A chronic 28 d toxicity study carried out against the mysid shrimp (Mysidopsis bahia) under flow-through conditions was submitted in summary form. Duplicate aquaria containing 30 shrimps were continuously exposed to mean measured concentrations of 0.65, 0.83, 1.2, 3.0 and 5.7 g ai l-1, each prepared with natural filtered seawater and solvent (acetone) carrier. Duplicate dilution water-only and solvent controls were also prepared and ran concurrently. No treatment related effects on adult survival, behaviour or growth were observed during the study. However, there were effects on reproduction with a significant reduction in the number of offspring per female reported for 1.2 g ai l-1 and above. Based on the results of this study the MATC of technical chlorothalonil against mysid shrimp was > 0.83 but < 1.2 g ai l-1. The 28 d NOEC for mysid shrimp chronically exposed to chlorothalonil was 0.83 g ai l-1 (Unpublished, 1995b).

6.4
6.4.1

ACUTE TOXICITY TO FISH

FRESHWATER FISH

In 1992, an acute toxicity test to determine the toxicity of a chlorothalonil formulation (54 % ai) to bluegill sunfish (Lepomis macrochirus) under flow-through conditions was carried out following FIFRA guideline 72-1. The study was also conducted to GLP. The test was conducted using a constant flow serial diluter, a temperature controlled water bath (22 1C) and a set of 14 exposure aquaria. The system was designed to provide nominal test concentrations of 108, 64.8, 38.9, 23.2, 14.04, 8.6 and 0 (dilution water-only control) g a.i l-1 to duplicate aquaria. The dilution water used was well-water with a total hardness 30 - 36 mg CaCO3 l-1. Each aquarium was maintained at a constant test volume of 11 l, with 6.5 volume replacements per day. The test aquaria were not aerated and were maintained in a 16L:8D photoperiod, with transition periods to avoid rapid shifts between light and dark. Ten fish were placed in each aquarium, giving a biomass loading of 0.054 g l-1, and mortalities were recorded and removed daily. Water quality parameters (pH, O2 and C) were measured daily, and samples of the test solutions were taken from each replicate on day 0 and 4 of the test period and chlorothalonil concentrations analysed by GC. The water quality 78

parameters were reported to have been unaffected by the concentrations of chlorothalonil formulation used, and that they remained within limits acceptable to the survival of bluegill sunfish. Measured concentrations of chlorothalonil were consistent between replicates and sampling intervals. The resulting mean measured concentrations were 64.8, 50.76, 27, 15.12, 8.1 and 3.9 g a.i l-1, and recovery of quality control samples was reported to have been 99.5 %. Following 24 h exposure 100 % mortality was observed among fish exposed to the highest mean measured concentration tested (64.8 g chlorothalonil l-1). At test termination (96 h), mortality of 95 % was observed among fish exposed to the mean measured test concentration of 50.76 g chlorothalonil l-1. Mortalities of 15, 0, 5 and 5 % were observed among fish exposed to mean measured test concentration of 27, 15.12, 8.1 and 3.9 g chlorothalonil l-1 respectively. FIFRA guidelines recognise that responses less than or equal to 10 % in a control population are within the expected range of naturally occurring variability. Therefore, the mortalities (5%) observed in the lower treatment levels (8.1 and 3.9 g ai l-1) were not wholly considered to have been as a result of exposure to chlorothalonil. The LC50 values estimated by non-linear interpolation, and the confidence intervals calculated by binomial probability, are presented in Table 6.3. Table 6.3: Acute Toxicity of Chlorothalonil to Bluegill Sunfish LC50 g ai l (95% C.L) 40.50 (27.0-50.8)
-1

48 h 96 h

35.10 (27.0-50.8)

Sub-lethal effects were observed (e.g. lethargy, partial loss of equilibrium) among all the surviving fish exposed to test concentrations > 27 g chlorothalonil l-1. Therefore, the NOEC for bluegill sunfish was determined to have been 27 g ai l-1 based on the lack of mortality and sub-lethal effects observed at this treatment level (Unpublished, 1992b). In 1992, an acute toxicity test to determine the toxicity of a chlorothalonil formulation (54.5 % ai) to rainbow trout (Oncorhynchus mykiss) under flow-through conditions was carried out following FIFRA guideline 72-1. The study was also conducted to GLP. The exposure system, an intermittent flow proportional diluter, was in operation for 4 d prior to test initiation to allow equilibration of the test material in the whole system and test aquaria. The system was set up to supply duplicate test aquaria with nominal test concentrations of 8.72, 14.17, 23.44, 39.24 and 65.40 g ai l-1, dilution water-only (control) and a chlorothalonil-free formulation (formulation control). The dilution water used was wellwater with a total hardness of 34-38 mg CaCO3 l-1 and a pH range of 7.2-7.3. The test solutions were not aerated, and the flow rate gave 7.2 volume replacements in 24 h.

79

Ten rainbow trout, with mean lengths and weights of 39 mm and 0.56 g respectively, were placed in each aquarium. All aquaria were held in temperature controlled water baths to maintain the test media at 12 1C, with a photoperiod of 16L:8D. Observations for immobility, sub-lethal effects (erratic swimming, lethargy etc.,), and physical characteristics of the test solution (e.g. precipitates) were made at test initiation and on each day thereafter. All mortalities were recorded and removed. Water quality parameters; hardness, DO, temperature and pH were measured daily in each replicate. Samples were taken from each test aquarium at 0 h (test initiation), 48 and 96 h, and analysed for chlorothalonil by GC. The water quality of the exposure solutions was considered to have been unaffected by the concentrations of chlorothalonil and remained within acceptable ranges for rainbow trout survival; DO, 89 - 94 % saturation; pH 7.0 - 7.2; temperature 12 C; total hardness 38 - 40 mg CaCO3 l-1. The mean measured test concentrations were reported as 6.54, 10.36, 15.81, 26.71 and 48.51 g ai l-1, which were 73 % of the nominal concentrations (mean recovery 101 %). Treatment related effects were only observed in the two highest test concentrations. After 48 h exposure mortalities (85 %) were only recorded in the highest measured test concentration of 48.51 g ai l-1. After 72 h, 100 % mortality was recorded for 48.51 g ai l-1, and 5 % for 26.71 g ai l-1 which increased to 15 % mortality after 96 h exposure. The LC50 values for chlorothalonil, calculated by non-linear interpolation, are presented in Table 6.4. Table 6.4: Acute Toxicity of Chlorothalonil to Rainbow Trout. LC50 g ai l (95 % C.L) 39.24 (26.71 - 48.51)
-1

48 h 96 h

33.25 (26.71 - 48.51)

Sub-lethal effects, e.g. loss of equilibrium, were also reported for the surviving fish exposed for 48 h to the top two chlorothalonil concentrations. At 15.81 g ai l-1, no mortalities were reported but several fish exhibited darkened pigmentation. This was not considered by the study author to be biologically significant or an indicator of an adverse response to toxic material, and the NOEC was reported as 15.81 g ai l-1 (Unpublished, 1992c). Further acute toxicity studies of technical chlorothalonil (>90%) against a number of different fish species were submitted in summary form. For the purposes of this review the results are presented in Table 6.5. All studies were carried out under static conditions with no aeration, and the concentrations were nominal, no analysis was performed.

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Table 6.5: Summary of Acute Toxicity of Technical Chlorothalonil to Freshwater Fish. Fish sp. Bluegill sunfish Lepomis macrochirus Channel catfish Ictalurus punctatus Rainbow Trout Oncorhynchus mykiss Species habitat warm water warm water cold water 96 h LC50 g ai l-1 (95 % C.L) 60 (50 - 75) 43 (26 - 70) 47 (32 - 68) NOEC g ai l-1 6 16 < 8.8

The summary report concluded from this data that chlorothalonil was highly toxic to fish and that there was little difference between the species of fish tested but that the estuarine/marine species tested appeared to be slightly more sensitive (Unpublished, 1995b). 6.4.2 ESTUARINE/MARINE FISH

An acute toxicity study carried out to determine the toxicity of technical chlorothalonil (>90%) against the sheepshead minnow (Cyprinodon variegatus) was submitted in summary form. The test was carried out under static conditions with no aeration, and the concentrations were nominal, no analysis was performed. The test was carried out using natural filtered seawater and a solvent carrier. Duplicate vessels containing 10 fish were run at each nominal concentration (20, 30, 45, 65 and 90 g ai l-1), along with duplicate dilution water-only and solvent control vessels. The general condition of the fish and mortalities were recorded every 24 throughout the 96 h exposure period. The 96 h LC50 value for sheepshead minnow exposed to technical chlorothalonil was 32 g ai l-1, with 95 % confidence intervals of 30 to 36 g ai l-1. The NOEC for this study was reported as 20 g ai l-1 (Unpublished, 1995b).

6.5
6.5.1

CHRONIC TOXICITY TO FISH

FRESHWATER FISH

A chronic study was carried out to investigate the effects of technical chlorothalonil on fathead minnow (Pimephales promelas) exposed during its early life, juvenile growth and reproductive stages. A brief summary of the study was submitted. The study was carried out under flow-through conditions without aeration throughout a complete (egg - egg) life cycle. The fish were exposed to mean measured test concentrations of 0.6, 1.4, 3.0, 6.5 and 16 g ai l-1 plus a dilution water-only and a solvent (acetone) control. No significant effects were reported for either generation exposed to 3.0 g ai l-1. Reproductive success of the first generation (number of eggs per spawn) was reduced at 6.5 g ai l-1, but no other treatment related effects were observed. Significantly reduced hatchability was reported noted for the second generation exposed to 6.5 g ai l-1, and no eggs were hatched from the fish exposed to 16 g ai l-1. The survival and growth rates of fry exposed to 6.5 g ai l-1 were not affected. Based on these results the report stated that the MATC based on the mean measured values was > 3 but less than 6.5 g ai l-1. The NOEC in fathead minnows chronically exposed

81

to technical chlorothalonil for two consecutive generations was given as 3 g ai l-1 (Unpublished, 1995b).

6.6

SUB-LETHAL EFFECTS IN FISH

Additional studies have been summarised by the HSE from the available literature studies investigating the sub-lethal effects of acute exposure to chlorothalonil. Gallagher et al, (1992a) investigated the toxicity of chlorothalonil on the channel catfish (Ictalurus punctatus). The paper reported a limit of solubility for chlorothalonil of 600 mg l-1 and stated that chlorothalonil was highly lipophilic. A static 96 h LC50 test was carried out using six 50 l aquaria, each containing five fish, with reported water characteristics of; temperature 23 C, pH 7.0 - 7.2, dissolved oxygen 5.0 - 6.0 mg l-1, total hardness 30 mg CaCO3 l-1. Chlorothalonil, dissolved in acetone, was added to five of the aquaria at concentrations ranging between 10 and 540 g l-1. The remaining group of five fish served as the control receiving only 2 ml of acetone. Water was changed and the fish redosed daily and the number of dead were noted and removed after 24, 48, 72 and 96 h exposure. Gross morphological and behavioural changes were also noted throughout the experiment, non were reported. The 96 hr LC50 for chlorothalonil was calculated by the BASIC microcomputer program and was reported as 52 g ai l-1. Davies (1987) investigated the various sub-lethal changes in anatomical, physiological and behavioural aspects of the respiratory system in rainbow trout (Salmo gairdneri now Oncorhynchus mykiss) following exposure to chlorothalonil. The relationship between LC50 responses and oxygen concentration was also examined. Fish were exposed to a series of chlorothalonil concentrations under flow-through conditions of either 8 or 5 mg oxygen l-1. During exposure blood samples and whole fish samples were removed for hematocrit and muscle lactate analysis. Observations of the ventilation rate in fish exposed to between 10 and 310 mg chlorothalonil l-1 were also carried out for up to 72 h. Oxygen concentrations of 5 mg l-1 significantly depressed the 96 h LC50 from 17.1 to 10.5 mg l-1 (P < 0.05) indicating a strong dependence of chlorothalonil LC50 on oxygen concentrations. Blood hematocrit values were seen to decrease in proportion to both concentration of chlorothalonil and exposure duration, and were accompanied by haemolysis. The decrease was most significant at concentrations > 20 g l-1 and this decrease was seen after only 24 h exposure to chlorothalonil. A similar response was seen in fish hematocrit levels exposed to increasing concentrations of the hydrolysis product DAC-3701, although the differences were not significant, and no haemolysis was reported. The reduced hematocrit observed following chlorothalonil exposure lead to acute anaemia which was accompanied by haemolysis. However, the effects on the hematocrit were said to be in general only transitory in response to stress induction and were compensated for eventually by physiological adaptation. Levels of muscle lactate decreased during the exposure experiments, therefore, it was shown that exposure to lethal levels of chlorothalonil did not result in acute tissue hypoxia. Exposure to high concentrations of chlorothalonil resulted in a dramatic increase in the ventilatory frequency which increased with increased exposure concentration. Gross body movements also increased dramatically on exposure to all test concentrations, accompanied

82

with bursts of very rapid ventilation, indicating acute stress. The maximal response was seen at 200 - 310 mg ai l-1. A minimum threshold level for response was reported to have been at 30 g l-1, where one in eight fish responded in a 2 hr exposure period. The paper quoted Long and Siegel (1975) who had showed that chlorothalonil inhibited key respiratory enzymes, explaining the increased sensitivity to chlorothalonil at a lower oxygen concentration. Prolonged exposure resulted also in gill damage, reducing the diffusing capacity. It should be noted that although the drop to 5 mg oxygen l-1 caused an increased toxic effect in response to chlorothalonil exposure by a factor of two - the oxygen concentration itself although at the lower end of the optimum conditions was not considered detrimental to salmonid sp. and stress symptoms were not observed in the control fish. In fact this is a common level in natural summer conditions. The authors concluded that environmental levels of 1 - 5 mg l-1 may have serious effects on the gill function if exposed for long time periods, or acute effects could be seen under conditions of low oxygen. However, chlorothalonil was not considered by the authors to be a persistent pesticide, as it would readily bind to suspended materials, and subsequently be removed from the environment by biodegradation. Gallagher et al, (1992b) investigated the in vivo toxicity and sub-lethal effects of chlorothalonil on the channel catfish (Ictalurus punctatus). Following exposure to chlorothalonil concentrations ranging between 10 and 540 mg l-1 for 96 h all surviving fish were bled by cardiac puncture and killed. To determine the sub-lethal target organ toxicity, four groups of six fish were exposed to chlorothalonil concentrations between 8 and 42 mg l1 , a fifth group of fish remained as the solvent control. Water was exchanged and the fish redosed daily. After 144 h (6 d) exposure all the fish were killed and the first gill arch, liver, posterior kidney, brain, intestine, spleen and heart were excised and fixed. Tissues from the control and highest dose group only were selected for the histopathological analysis due to the absence of behavioural signs elicited in the lower dose groups. Also the effects on glutathione (GSH) concentrations were investigated for the liver, gill and posterior kidney of catfish exposed to 13 g chlorothalonil l-1 for 6, 24 and 72 h. Exposure to a sub-lethal concentration of chlorothalonil for 6 days resulted in necrosis of the intestinal epithelial lining in 57 % of the treated fish. However, no treatment related pathological lesions were observed in the liver, gills, posterior kidney, brain, spleen or heart. Catfish exposed to 13 g chlorothalonil l-1 had increased tissue GSH concentrations in the liver, posterior kidney and gills, suggesting a protective role of tissue GSH towards chlorothalonil exposure. Davies et al, (1994) investigated the sub-lethal responses of various species of fish and crustacea to short term, low concentrations exposures (10 or 20 d) of chlorothalonil. Six, 40 l partitioned glass tanks (80 % covered in black plastic) were set up for each organism tested, with 10 to 15 fish and 20 to 30 crustaceans placed in each tank. Five test concentrations (2 to 10 mg l-1) and one solvent control was used for each test organism. Tanks were supplied with a flow-through of medium at a constant temperature (12 - 15 C) and rate sufficient to sustain stable chlorothalonil concentrations and maintain a high oxygen level (> 80 % saturation) without supplementary aeration. Natural US water (pH 6.5 - 7 and dissolved oxygen > 95 %) was used for the dilution medium. Test concentrations were sampled during the experiments and analysed by GC-ECD.

83

Several parameters were examined; hematocrit, leucocrit, plasma glucose, plasma protein, plasma chloride, muscle RNA and DNA, brain and muscle acetyl cholinesterase (AChE) activity, hepatic glutathione-S-tranferase activity, hepatic glutathione and somatic growth. Only the parameters where significant results were observed were reported by the authors. The acute LC50 results for chlorothalonil are presented in Table 6.6. Table 6.6: Summary of LC50 Values of Freshwater Fish and Invertebrates Exposed to Chlorothalonil Species Oncorhynchus mykiss Pseudaphritis urvillii Galaxias maculatus Paratya australiensis Astacopsis gouldi Colubotelson chiltoni Neoniphargus sp. A Test duration (d) 10 10 10 4 7 4 7 4 7 4 7 LC50 g l-1 (confidence limits) > 8.2 > 8.2 > 8.2 16.0 (14.4 - 17.9) 10.9 (9.0 - 13.1) 12.0 (7.9 - 18.1) 3.6 (2.1 - 6.0) >40 >40 >40 >40

The results show that chlorothalonil appears to be slightly more toxic to fish than crustacean under the reported conditions of the test. Both Oncorhynchus mykiss and Pseudaphritis urvillii demonstrated dose dependant elevations in hepatic GSH levels following exposure to sub-lethal concentrations of 1.4 and 8.2 g l-1 respectively. Hepatic glutathione-S-tranferase (GST) activity was elevated in fish exposed to concentrations > 0.8 g chlorothalonil l-1 but this was not significant (P = 0.005) until 1.4 g chlorothalonil l-1. Respiratory activity was also significantly higher for all fish exposed to concentrations 0.3 g ai l-1, the response being dose dependent and increasing by a factor of two over the exposure concentration range. Exposure to chlorothalonil induced a depression in muscle or whole body AChE in all the crustacean species tested with the exception of Neoniphargus sp. A, which was not used for this analysis. Whole body GSH and GST elevations were reported in only one crustacean; Paratya australiensis exposed to 0.3 and 1.8 g l-1 respectively. From the data generated from this study the threshold concentration for significant effects was set at 0.3 g l-1 for chlorothalonil. The author commented that the elevation in GST activity indicated that chlorothalonil induced a physiological response in the fish and crustacean species tested. This was a specific 84

functional response as opposed to a stress-typed response more commonly seen with other pesticides exposure. The induction of this phase II detoxification enzyme is related to chlorothalonils ability to catalyse the conjugation of electrophilic molecules. Chlorothalonil has several reactive electrophilic halogens (chlorine) in the molecule that may lead to direct conjugation with glutathione. Gallagher et al, (1992b) investigated the protective role of GSH in liver and gills of channel catfish (Ictalurus punctatus) exposed to chlorothalonil. In an initial experiment the acute toxicity of chlorothalonil was enhanced threefold after channel catfish were depleted of liver and gill GSH by L-buthionine S,R-sulfoximine and diethyl maleate. This indicated that GSH plays a critical role in mitigating acute chlorothalonil toxicity in this species. In a subsequent experiment, sub-lethal exposure to chlorothalonil elicited significant increases (P < 0.05) in hepatic and gill GSH and cysteine concentrations after 72 and 144 h exposure. Also, an apparent induction of hepatic gammaglutamylcysteine synthetase (GCS) activity was also noted after 144 h exposure. This was also noted in the gills but was not significant (P > 0.05). The results indicated that the mechanism of GSH induction in the liver and the gills of catfish exposed to chlorothalonil involves increased cysteine uptake and also increased GCS activity. These results were considered by the authors to support their hypothesis that simultaneous exposure to multiple environmental chemicals that utilise or deplete tissue GSH may predispose fish to acute toxicity.

6.7

TOXICITY TO BIRDS

Several toxicity studies have been submitted in summary form. These studies suggest that chlorothalonil is of very low toxicity to birds (See Table 6.7). Table 6.7: Summary of Chlorothalonil Toxicity to Birds Test type Acute Oral Species Mallard duck Japanese quail Dietary Mallard duck Bobwhite quail LD50 /LC50 > 4640 mg kg-1 > 2000 mg kg-1 > 10000 ppm > 10000 ppm NOEL 2150 mg kg-1 2000 mg kg-1 5620 ppm 5620 ppm

In reproduction studies with the mallard duck and the bobwhite quail, treatment related reductions in egg production, hatching and offspring survival were reported but these levels were extremely high levels (NOEC > 1000 and > 500 ppm respectively) (Unpublished, 1995b).

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6.8 METABOLITE TOXICITY TO AQUATIC ORGANISMS

SDS-3701 has been shown to occur in natural sediment-water systems but only as a minor metabolite. However, within the summary data submitted, several studies have been reported where the toxicity of this metabolite has been assessed. All the studies were carried out under static conditions using nominal concentrations only. The reported endpoints have been presented in Table 6.8.

Table 6.8: Acute Toxicity of Metabolite SDS-3701 to Aquatic Organisms

Species Daphnia magna Bluegill sunfish Bluegill sunfish

End point (g l-1) 48 h EC50 - 25600 NOEC -10000 96 h LC50 - 16000 NOEC - 6000 96 h LC50 - 45000 NOEC - 18000

Unlike technical chlorothalonil, which was shown to be highly toxic to fish and aquatic invertebrates, SDS-3701 was found to be only slightly toxic in comparison (Unpublished, 1995b).

6.8

PUBLISHED REVIEW

In addition to the above company and literature data a comprehensive review of chlorothalonil for the purpose of assessing the risks to the aquatic compartment has been produced in Canada by Caux et al, (1996). This review, which has been used to produce the water quality guidelines for chlorothalonil in Canada (including the drinking water supply, freshwater and marine life, crop irrigation, livestock watering, recreational aesthetics and industrial waters) has been summarised and below the relevant ecotoxicological endpoints covered by the document have been tabulated in Tables 6.9 - 6.14. The specific references for these end points are available from Caux et al, (1996).

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Table 6.9: Toxicity of Chlorothalonil and Metabolite DS-3701 to Freshwater Algae Species Chlorothalonil Selenastrum subspicatus DS-3701 Selenastrum subspicatus S, N Log phase growth 7 d IC50 - 33700 100 S, N Log phase growth 7 d IC50 - 8500 96 Test type Life Stage End point (g l-1) % ai

Key: S - static; R - static renewal; F - flow through; N - nominal concentration; IC50 - concentration resulting in 50% growth inhibition. The data presented in Table 6.9. suggests that the ai is almost five times more toxic to freshwater algae than the major soil metabolite DS-3701. However, it should be noted that the IC50 quoted exceeded the solubility of chlorothalonil, and so it may be concluded that chlorothalonil is of low toxicity to freshwater algae under the conditions tested. Table 6.10. presents data which also suggests that the toxicity of the metabolite DS-3701 to freshwater invertebrates is significantly lower than that of the ai, which can be considered highly toxic from the data reported. Table 6.10: Species Chlorothalonil Daphnia magna Toxicity of Chlorothalonil and Metabolite DS-3701 to Freshwater Invertebrates Test type S, M Life Stage < 24 h End point (g l-1) 48 h EC50 - 97 (unfed) - 172 (fed) 48 h EC50 - 70 % ai 40

S, N < 24 h 96 Daphnia magna 99.8 S, N < 24 h 48 h EC50 - 120 Daphnia magna Freshwater F, M 0.05 - 0.15g 96 h EC50 - 16 >98 shrimp 7 d EC50 - 10.9 Freshwater F, M 0.13g 96 h EC50 - 13 >98 shrimp 7 d EC50 - 3.6 Isopod F, M NR 7 d EC50 - >40 >98 Amphipod F, M NR 7 d EC50 - >40 >98 DS-3701 99 S, N < 24 h 48 h EC50 - 26,000 Daphnia magna Key: S - static; R - static renewal; F - flow through; N - nominal concentration; M - measured concentrations; NR - not reported.

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The data presented in Table 6.11. suggests that the toxicity of chlorothalonil is at least 50 times more than that reported for DS-3701. In comparing the freshwater and marine invertebrate data, it would appear that the marine species are less sensitive to acute chlorothalonil exposure. However, most of the data quoted in Table 6.11 was based on actual concentrations, therefore, there is the possibility that this data underestimates the actual toxicity levels. Table 6.11: Toxicity of Chlorothalonil and Metabolite DS-3701 to Estuarine/Marine Invertebrates Life Stage 25 - 50 mm End point (g l-1) 96 h EC50 - 7.3 (reduced shell growth) 96 h EC50 - 165 96 h EC50 - 320 20 minb NOEC - >128 (adverse fertilisation effect) % ai 98.2

Species Test type Chlorothalonil Eastern F, M oyster Pink shrimp Pink Shrimp Purple sea urchin DS-3701 Purple sea urchin S, N F, N R, N

Adult Juvenile Gametes

100 > 90 96

20 minb NOEC - >6400 100 (adverse fertilisation effect) Key: S - static; R - static renewal; F - flow through; N - nominal concentration; M - measured concentrations; b - sperm exposed for 10 min. followed by an additional 10 min. for sperm and eggs. R, N Gametes

Many chlorothalonil toxicity studies have been reported for the freshwater fish, rainbow trout, and the results are presented in Table 6.12. The acute toxicity levels reported were in the range of 10.5 - 76 g l-1 (96 h LC50) which under the conditions tested suggests that chlorothalonil would be highly toxic to fish. This statement is further supported considering the reported 10 d LOEC of 1.4 g ai l-1 following chronic exposure to chlorothalonil.

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Table 6.12: Toxicity of Chlorothalonil to Rainbow Trout Test type S, M S, N R, N F, M F, M S, M S, N F, M Life Stage 3.5 - 4.0 g Fry Fry 6 - 11 g 6 - 11 g 6 - 11 g 0.64 - 1.54 g 1.1 - 2. 5g End point (g l-1) 96 h LC50 - 76 96 h LC50 - 57 28 d LC50 - 54 24 h LC50 - 40.2 48 h LC50 - 18.8 96 h LC50 - 10.5 96 h LC50 - 18 96 h LC50 - 47 % ai 98 96 96 99 99 99 96

10 d 98 LOEC - 1.4 NOEC - 0.8 (elevated glutathione) Key: S - static; R - static renewal; F - flow through; N nominal concentration; M - measured concentrations; Further studies involving toxicity studies with the major soil metabolite DS-3701 against rainbow trout are presented in Table 6.13. These data suggest that toxicity of the metabolite is far less than that previously demonstrated with chlorothalonil under conditions of either acute or chronic exposure. Table 6.13: Toxicity of Metabolite DS-3701 to Rainbow Trout Test type R, N R, N R, N S, N Life Stage Fry Fry Fry Fry End point (g l-1) 28 d LC50 - 3400 28 d LC50 - 57 28 d LC50 - 32 96 h LC50 - 9200 % ai 100 100 100 100

Key: S - static; R - static renewal; N - nominal concentration; M - measured concentrations; Marine fish have been studied to a lesser degree, but the data presented in Table 6.14., further confirms that chlorothalonil is of high toxicity to fish, but that the metabolite is significantly less toxic. The data here also suggests there was little difference in the sensitivity between the marine and freshwater species tested.

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Table 6.14: Toxicity of Chlorothalonil and Metabolite DS-3701 to Marine Fish Species Chlorothalonil Threespined stickleback Threespined stickleback Sheepshead minnow DS-3701 S, M S, M S, N 0.3 g 3.4 cm NR 3 - 7d 96 h LC50 - < 73 96 h LC50 - 27 96 h LC50 - 320 40 96 >90 Test type Life Stage End point (g l-1) % ai

Threespined S, M NR 96 h LC50 - 4700 100 stickleback Key: S - static; R - static renewal; F - flow through; N - nominal concentration; M - measured concentrations.

6.9

SUMMARY OF ECOTOXICOLOGY

From the data submitted by the supporting company, chlorothalonil was shown to be of low toxicity to the freshwater algae Selenastrum subspicatus with an 5 d EC50 of 0.21 mg ai l-1. This was supported by the reviewed literature which reported a 7 d IC50 of 8.5 mg ai l-1 for the same species Selenastrum subspicatus. Acute exposure of invertebrates, Daphnia magna, Penaeus duorarum and Crassostrea virginica, to chlorothalonil demonstrated significant toxicity but did not highlight major differences in sensitivities between freshwater and marine species with acute EC50 values of 97.20 (48 h), 165 (96 h) and 7.3 (96 h) g ai l-1 respectively. Following chronic exposure to chlorothalonil, marine invertebrate demonstrated greater sensitivity than freshwater species, i.e. the Daphnia magna 21 d NOEC of 35 g ai l-1 was significantly higher than the 28 d NOEC of 0.83 g ai l-1 reported for Mysidopsis bahia. In freshwater fish, acute exposure to chlorothalonil resulted in high toxicity with similar LC50 values reported for the bluegill sunfish and the rainbow trout at measured concentrations of 35.10 and 33.25 g ai l-1 respectively. A 96 h LC50 of 32 g ai l-1 reported for the marine fish, sheepshead minnow, suggested that acute chlorothalonil toxicity was not significantly different to that reported for freshwater species. Under chronic conditions, a NOEC of 3 g ai l-1 was reported for the freshwater species, fathead minnow. No chronic data for marine species were submitted. The review of literature studies demonstrated that there were overlaps in the reported ranges of chlorothalonil toxicity to a wide range of fish and invertebrate species, including the submitted company data. However, the lowest NOEC was reported for rainbow trout (10 d, 0.8 g ai l-1) suggested that chlorothalonil was potentially more toxic than the company data had demonstrated. The review of available literature where the sub-lethal effects of chlorothalonil against several aquatic species had been investigated, demonstrated that chlorothalonil could elicit significant toxic effects at quite low concentrations. Davies (1987) reported that reduced

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oxygen levels increased the acute toxic response to chlorothalonil, and on further investigation reported two effects of chlorothalonil which explained this effect. Chlorothalonil was shown to trigger respiratory stress (increased ventilation) after only 2 h exposure to 30 g ai l-1, and that the hematocrit levels in surviving fish were elevated and accompanied by haemolysis. However, the author concluded that this response was only transitory and if prolonged exposure occurred physiological adaptation would compensate for it. This response was not seen in the presence of the primary metabolite, DAC-3701, which may further explain its reduced toxicity previously discussed. Davies et al (1994), investigated sub-lethal responses in fish and crustacea. In fish, at exposure levels of 1.4 g ai l-1 increased hepatic GSH activity predominated, and respiratory stress was noted at exposures exposed to 0.3 g ai l-1. However, in crustacea exposed to 0.3 g ai l-1 decreases in muscle or whole body AChE were significant. The suggestion is therefore that the differences in toxic profiles between fish and crustacea are not merely dose response but that the target of chlorothalonils action is different. Summary data presented by the company suggested that chlorothalonil was of low toxicity to birds under the conditions tested.

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7 ENVIRONMENTAL RISK ASSESSMENT

7.1 ENVIRONMENTAL HAZARD PROFILE

Company reported leaching rates varied from 0.99 to 3.18 g cm-2 d-1. In the aquatic environment chlorothalonil biodegraded rapidly with half-lives of < 2 h in aerobic marine and freshwater-sediment systems. Two major metabolites (SDS-67042 and SDS-67042 sulfoxide) were reported in both systems, however, their persistence was not clear. Chlorothalonil was shown to degrade in soil, but this was not rapid with half-lives of between 8 and 36.5 d. One major metabolite was reported following degradation in soil, SDS-3701, demonstrating a different degradation pathway to that under aquatic conditions. Chlorothalonil appeared to adsorb quite strongly to soil, but the degree to which it could then desorb was not clearly established. Chlorothalonil residues did bioaccumulate in fish, with a maximum BCF of 2867, however, this was calculated for 14C and characterisation studies suggested that the parent a.i was largely metabolised. Depuration was also shown to be relatively rapid (half-life < 14 d). Chlorothalonil was shown to be highly toxic to aquatic species. The species most sensitive to chronic exposure was the mysid shrimp with a 28 d NOEC of 0.83 g ai l-1 based on reproductive effects. Sub-lethal effects following short and long-term exposure have been documented, however, the significance of the effects on respiratory rate and glutathione-Stranferase activity are not clear.

7.2

RISK ASSESSMENT STRATEGY

The risk assessment has been concentrated on the marine environment since the data available are predominantly for the use of antifouling products (AFPs) in estuarine and coastal areas, although the risk to freshwater environments has not been precluded. However, the strategy for assessing risk to the marine environment is less well developed than for terrestrial or freshwater. Therefore, the risk assessment strategy adopted by HSE for the purposes of the current review has been presented in a separate document Environmental Risk Assessment of Booster Biocides (ACP [2002]). This document presents a comprehensive and comparative risk assessment for all approved booster biocides and has been endorsed by an expert Ad-hoc Environmental Panel. For the marine environment, three distinct areas were identified; estuarine (including marinas and harbours), shallow coastal seas and deep ocean. Within each area considerations of the following compartments; sediment (including suspended sediment), water and associated biota are required. Therefore the primary objective of this risk assessment has been to establish the likelihood of chlorothalonil reaching (or having reached) a concentration in the aquatic environment which would adversely affect some component of that environment. In order to achieve this objective, measured environmental concentrations (MECs), predicted environmental concentrations (PECs) and predicted no effect levels (PNECs) have been derived.

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Below are the main points of the risk assessment detailed in Environmental Risk Assessment of Booster Biocides (ACP [2002]) concerned with the use of AFPs containing chlorothalonil, however, reference to the complete document is advised. 7.2.1 ENVIRONMENTAL CONCENTRATIONS

The direct and indirect exposure of the aquatic environment to chlorothalonil as a result of AFP use are detailed fully in Environmental Risk Assessment of Booster Biocides (see Section 7.2). Data have not been made available to address the direct and indirect inputs to the soil, and aquatic compartments as a result of AFPs use (application and removal stages). Further to this, HSE have no data regarding the direct exposure of STP (sewage treatment processes), or the subsequent emissions to surface waters. However, a survey carried out by the Environment Agency (EA) in 1998 demonstrated that there was a wide range of use patterns and work practices regarding the application and removal of AFPs. The survey suggested that emissions via drains would be low, since direct exposure of the surrounding water body (marina/harbour) was more likely, with the majority of boat maintenance taking place at the waters edge. Although the survey did state that where removal by sandblasting was undertaken (usually by boatyards) appropriate waste disposal methods were reported. The contamination of aquatic sediments from direct emissions or via particulate inputs (contaminated soil) is an important route to consider, since the potential to contribute significantly to the environmental loading of the booster biocides exists through remobilisation. Where strongly adsorbed ais are demonstrated, in the aquatic environment mechanical remobilisation, or sediment redistribution via dredging are likely. Therefore, exposure scenarios other than direct exposure of the aquatic compartment (i.e. indirect exposure of the aquatic environment, direct exposure of the soil and STP) have not been considered further at this stage with insufficient data available to fully address the risks. The potential scale of direct exposure to the aquatic compartment may also be considered to outweigh the additional areas of concern at this time. Usage information was requested from all the current approval holders for chlorothalonil, however, the information received was too limited to allow for any accurate assessment of the actual amounts used of AFPs containing chlorothalonil. Therefore, the results of a survey conducted by the EA in 1998 were used instead. The EA survey demonstrated that chlorothalonil was not currently used to any significant level (< 3 % pleasure crafts were treated with chlorothalonil). Seawater monitoring data provided by CEFAS (Centre for Environment, Fisheries & Aquaculture Science) on behalf of a DETR (Department of Environment Transport and the Regions) commissioned monitoring program in 1998, demonstrated that chlorothalonil was not detected at any of the sites. No sediment analysis was conducted for chlorothalonil. However, the current monitoring data could only ever represent the usage levels for 1998, and predictions of maximum PECs were considered necessary by the Ad-hoc Environmental Panel since post approval usage cannot be controlled. Therefore, PEC data based on 100 % usage (all vessels treated) of chlorothalonil AFPs were predicted using a model developed as part of a HSE/EA commissioned research project. The model, REMA (Regulatory Environmental Modelling of Antifoulants), is a steady-state QWASI (quantitative water air sediment interface) model, designed to predict concentrations of biocides in both the water and sediment compartments of estuaries/marinas/harbours. The model is based upon for four real estuary scenarios in the UK, for which the model has been successfully validated. The model inputs used for chlorothalonil are presented in Table 7.1, and a summary of the results in Table 7.2.

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Table 7.1: REMA model input parameters for chlorothalonil Input Melting point Molecular mass Vapour pressure Solubility Log Koc Sediment half-life (h); Hydrolysis Photolysis Biodegradation Water half-life (h) Hydrolysis Photolysis Biodegradation Leaching Usage (% boats treated) Value 250.5 C 265.9 0.076 mPa 0.6 mg l-1 3.6 10 000 000 10 000 000 2 10 000 000 1552.8 2 3.18 g cm-2 d-1 3 % and 100 % Company Company Pesticides Manual (20 C) Company Calculated from Kow (company data) Default (no data) Default (no data) Company, marine aerobic Default (none at pH 7) Company Company, marine aerobic Company, ASTM method Surrogate from EA 1998 Survey and maximum Source

Table 7.2: Summary of PEC calculations for chlorothalonil in estuaries and open marinas Usage (%) 3 Site PECwater (ng ai l-1) 0.4 ( 0.67) 9.3 ( 5.6) 12.1 ( 22.2) 306.6 ( 184.6) PECsediment (g ai g-1) 5.0x10-8 ( 8.3x10-8) 9.5x10-7 ( 6.7x10-7) 1.7x10-6 ( 2.8x10-6) 3.2x10-5 ( 2.2x10-5)

Estuary* Open marina** 100 Estuary* Open marina** * n = 12, **n = 7

The predicted water concentrations presented above exceeded the limit of detection quoted by CEFAS (1 ng l-1) for marinas regardless of use level and in the estuary where the usage figure of 100 % was used. The mean PEC calculations for the sediment compartment were very low, and suggest that regardless of usage or location (marina/estuary) these would be below the limit of detection suggested for chlorothalonil by CEFAS (0.001 g ai g-1) although it may be possible to detect the levels presented in Table 7.2, at present there is no information available to confirm this. 7.2.2 PREDICTED NO EFFECT CONCENTRATIONS

Once released into the aquatic compartment, the chemical fate of the booster biocide will determine whether the toxic effect exerted is limited to the target organisms within a boundary layer of a painted surface or whether the ai persists and there is potential for

94

exposure to non-target organisms. Therefore, selection of key non-target organisms and likely duration of exposure is essential, but this is somewhat reliant on the availability of acceptable data for representative marine species. Chronic data endpoints have been selected as more appropriate for the purpose of a marine risk assessment following the use of booster biocides. This is because the inputs of booster biocides into the marine environment as a result of leaching from multiple point sources (treated surfaces) will be a continuous process. Even where high degradation is indicated from fate studies, the continuous input from leaching can still result in long-term exposure. This will only be mitigated if the leaching rate of an ai from a painted surface is significantly slower than the rate of degradation, and no toxic metabolites are produced. Comparisons between marine and freshwater chronic toxicity data for booster biocides has not demonstrated any differences in sensitivities. Therefore, in the absence of chronic marine data, freshwater data would be acceptable. Further to this, in considering the number and quality of tests available for the current review, the HSE have selected the most sensitive species, regardless of test medium. However, the introduction of assessment factors is required before deriving a PNEC from the available hazard data, which in the absence of additional or more appropriate data will provide a suitable safety margin for all marine organisms. The provision of assessment factors were made in accordance with the guidance detailed in the European Risk Assessment Technical Guidance Document (EURATGD, 1996), and those previously accepted by the ACP. The most sensitive chronic endpoint available for chlorothalonil was a 28 d NOEC based on reproduction of 0.83 g ai l-1 for the marine mysid shrimp. The data set submitted for chlorothalonil included acute toxicity for both marine invertebrates and fish, and chronic studies for algae (freshwater), invertebrates (freshwater and marine) and fish (freshwater). Additional marine studies were also available in published studies. Therefore, an assessment factor of 10 was considered appropriate, giving a PNEC of 0.083 g ai l-1 for the mysid shrimp. 7.2.3 RISK QUOTIENT

The PEC:PNEC calculations derived from the predicted data based on 100 % usage for chlorothalonil were considered to be the most appropriate for regulation. This approach was suggested and endorsed by the Ad-hoc Environmental Panel, for the primary reason that postapproval control of amounts used is not possible, and therefore a worst-case assumption is necessary. In addition to this the only usage data currently available are those regarding the number of pleasure crafts located in UK waters, therefore, the inputs from ships would be additional. Finally, no allowance has been made regarding the potential for additivity, synergism or antagonism between ais which will coexist in the aquatic environment as a result of leaching from AFPs, which the HSE considers as additional support for the 100 % worst case usage approach.

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Table 7.3: Summary of chlorothalonil PEC:PNEC ratios based on 100 % use on pleasure crafts Estuary PEC:PNEC Mean % ( STD) Exceedence* 0.2 ( 0.3) 0 Open marina PEC:PNEC Mean % ( STD) Exceedence* 3.7 ( 2.2) 100

* - % Exceedence is the percentage of single samples which were shown to exceed the MEC:PNEC quotient of 1, demonstrating that the measured concentration of booster biocides was greater than the no effect concentration and therefore indicating significant risk of adverse effects.

The summary data presented in Table 7.3 suggest that for chlorothalonil, the only area where unacceptable risk would be likely to occur would be in open marinas with 100 % of the data exceeding a PEC:PNEC ratio of 1.

7.3
7.3.1

SIGNIFICANCE OF RESULTS
WATER CONCENTRATIONS

The predicted data provided by the REMA model have allowed development of the risk assessment to take on board the maximum risk posed from the pleasure craft use of chlorothalonil AFPs in the UK. Unfortunately it has not been possible to predict the environmental concentrations resulting from the use of chlorothalonil on larger commercial vessels and ships. Therefore, the predicted concentrations at the 100 % use level (for pleasure crafts) are more likely to underestimate the environmental exposure resulting from AFPs use. The REMA model data have indicated that for chlorothalonil, use in AFPs may result in unacceptable exposure of open marina sites. Therefore, refinement to the model parameters or subsequent monitoring of representative areas are considered necessary to address the risk from use on pleasure crafts in the UK. In addition, outstanding data clarifications and metabolite data have been requested. Advice was sought from the ACP as to whether information should be requested regarding the projected use of chlorothalonil in AFPs suitable for larger vessels and deep sea use (see Environmental Risk Assessment of Booster Biocides Currently Approved for Use in UK Antifouling Products). 7.3.2 SEDIMENT TOXICITY

The PECsediment data derived by the REMA model suggest that the sediment compartment is not a concern for chlorothalonil. Therefore, no additional data to address chlorothalonils behaviour or toxicity in sediment have been requested.

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7.3.3

SECONDARY AND SUB-LETHAL ECOTOXICOLOGY DATA

Potential secondary effects as a result of fish bioconcentration of chlorothalonil are not considered to pose unacceptable risks from the available data. The potential for indirect or sub-lethal effects (i.e. endocrine disruption), should be considered in all cases where prolonged exposure to concentrations below those shown to elicit known toxic effects are likely. This consideration should remain until reliable data can be obtained which demonstrates that such effects do not occur. This is especially important when considering the use of booster biocides which result in direct environmental exposure. Other sub-lethal effects such as respiratory distress and stimulation of protective protein mechanisms have been reported. However, whilst the Ad-hoc Environmental Panel recognised that the significance of these data are difficult to determine, since assessments for endocrine disruption (and other sub-lethal effects) are as yet not available, they recommended that the potential for both these effects should not be ignored. Indeed, when the techniques become available the panel members agreed that persistent ais used in AFPs (or their metabolites) should be reconsidered. 7.3.4 METABOLITE FATE AND EFFECTS DATA

Degradation of a parent compound is often where the considerations for environmental risk end. However, risk assessments should be conducted for all reported persistent major (> 10 %) metabolites. Without consideration of the metabolites, a full assessment of the risk to the marine environment from the use of AFPs cannot be considered complete. Clarification has been requested for the metabolism of chlorothalonil in marine sediment-water systems, and where significant persistent metabolites are highlighted additional data will be requested. 7.3.5 MIXTURES OF SUBSTANCES

AFPs, usually contain a mixture of booster biocides with copper or TBT compounds. Therefore, the environment will be routinely exposed to such mixtures as a result of in situ leaching. Under the current regulations (COPR, 1986) AFPs are registered and approved on an ai basis, and mixtures of ais are not considered. Our general understanding of the possible effects arising from mixtures such as synergism, additivity and antagonism, is as yet not sufficiently developed. Also, when considering the number of AFPs and potential combinations of these in the field at any one time, the applicability of laboratory studies in gaining an understanding of effects in the field becomes so complex that a realistic assessment is unlikely. In addition to this, the chemistry of booster biocides in paint matrices is very complicated and caution would have to be applied in the interpretation of results. A simpler approach may be to consider the mode of action of the ais present in AFPs, for example chlorothalonil and other ais with known fungicidal action. However, in the absence of data, mixtures will not be considered further within this risk assessment with the assumption that assuming 100 % use of single ais takes at the very least additivity into account.

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7.4

DATA REQUIREMENTS

Ministers have agreed the following data requirements, which are to be submitted within a total of 3 years, with shorter timetables for some requirements as outlined below. The data requirements are based on data gaps or have been requested as a result of unacceptable high predictions of environmental exposure using the REMA model. Approval Holders must demonstrate to HSE how they are implementing the data requirements within 6 months. 1. 2. 3. Inhibition of microbial respiration in sewage sludge (core requirement) to be submitted within 12 months. Fish early-life stage test (sheepshead minnow preferable species) to be submitted within 12 months. Clarification that the leaching rate of the parent ai used for the risk assessment is representative of current products. This information is to be submitted within 12 months. (Repeated REMA calculations will be carried out by HSE with any new data supplied.) Should the leaching data submitted as part of requirement 3 fail to reduce the predicted environmental concentrations of chlorothalonil to acceptable levels (assuming 100 % use on pleasure vessels) a monitoring programme will be necessary. In the first instance, HSE should liaise with DETR, EA and English Nature and investigate the potential for further monitoring via the National Monitoring Programme or other organisations. Confirmation that chlorothalonil significantly degrades in marine sediment-water systems, and identification of any subsequent persistent metabolites to be submitted within 12 months. Once the significance and persistence of the metabolites have been established additional ecotoxicology, fate and behaviour data may be required in order to permit a risk assessment to the same level as that of the parent compound. The specific data required should be discussed with HSE at on completion of data requirement 5 and all subsequent data requirements submitted within a further 2 years. A study to address bioacumulation of chlorothalonil in shellfish, in order to address the potential risks to consumers via the food chain, should be submitted within 2 years.

4.

5.

6.

7.

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8 EFFICACY
8.1 BACKGROUND/INTRODUCTION
Two processes describing the fouling of an immersed surface by organisms are recognised in the literature, and are presented in two papers. The first of these papers describes a classical view of fouling, in which fouling is reported to occur as a successional process. A freshly immersed surface rapidly adsorbs various organic molecules to form a conditioning film. This film facilitates subsequent colonisation by such micro-organisms as bacteria and diatoms, which results in the formation of a slime layer. The microfouling slime layer is subsequently succeeded by macrofouling. This can be of two types, 'hard' and 'soft' fouling. Common soft-fouling organisms include algae, sponges, tunicates and hydroids. Common hard-fouling organisms include barnacles, mussels, clams and tubeworms. Larvae of fouling organisms are more diverse and numerous in coastal areas than in the open sea, therefore the fouling challenge is more intense in these areas (US Naval Institute, 1981). An alternative theory to succession has been offered by other workers. This is a much more complex process depending on the relative amount and type of fouling organism present. The different organisms are in dynamic equilibrium with the immersed surface, and in its absence, with flocculation of 'marine snow' (waste material, dead micro-organisms etc falling through the water column). There are a number of secondary driving forces and behavioural interactions in this model. For example, certain types of microbial fouling may actually inhibit the settlement of macrofouling organisms (Clare et al, 1992). Animal and weed fouling of vessel surfaces increases the drag on the hull. This results in increased fuel costs and a reduction in the vessel's speed and manoeuvrability. Beyond a certain level, fouling will require removal from the vessel by its owner during dry docking. The performance of antifouling products may be judged as the maximum specified time that a vessel can spend 'in service' before having to be dry docked, cleaned and recoated. For insurance purposes, commercial vessels have to be dry docked at least every 5 years for inspection and maintenance. Therefore, the ideal antifouling product would protect a vessel for this length of time, so that extra expense is not incurred as a result of more frequent dry docking. The period of time between repainting can, however, vary quite considerably depending upon the chosen antifouling product formulation, and the environment in which it is used. Various environments will present different fouling challenges. Parameters which will influence the service time of an antifouling product include:Trading patterns (coastal or deep sea, turn around time, speed of vessel etc.), Fouling conditions (warm or temperate water), Physico-chemical conditions of the sea water e.g. pH and temperature, Coating type and film thickness.

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8.2 ACTIVE SUBSTANCES/COATING TYPES USED IN ANTIFOULING PRODUCTS

Antifouling products can be broadly divided into two types: Those that contain tributyltin (TBT), and those that are TBT-free. TBT-free products typically contain copper or a copper compound such as copper (I) oxide (Cu2O), or copper (I) thiocyanate (CuSCN) as the principal biocide. Since some of the common algae such as Enteromorpha spp. and Amphora spp. are tolerant of copper, this active ingredient is 'boosted' or enhanced by the presence of one or more organic biocides such as Irgarol 1051 or dichlorophenyl dimethylurea (Diuron). These are usually algicides, but may, in addition, possess a wider spectrum of antifouling activity. Copper is also used to enhance the performance of TBT products because organisms such as Ectocarpus spp. and Achanthes spp. are tolerant of tributyltin oxide (TBTO) (Callow, 1990; Unpublished, 1994). The antifouling products can be further categorised into the following broad coating types:Soluble matrix (formerly known as conventional), Insoluble matrix (formerly known as contact leaching), TBT and TBT-free self polishing copolymer (formerly known as TBT and TBT-free ablative) The categorisation of coating types outlined above is very generalised. It should be noted that apart from the TBT self polishing co-polymer (SPC) products, the majority of other antifouling products do not necessarily rely on one single coating technology. Instead, composites of different technologies have been developed by antifouling formulators to suit customer specifications and environmental requirements. Further detail and descriptors for the individual coating types can be found in Appendix 2. The Antifouling Manufacturers Working Group of the European Confederation of Paint, Printers', Inks and Artists Colours Manufacturers Association (CEPE) have agreed maximum protection periods that can be expected for each antifouling coating type. The types of coating and maximum protection periods with respect to recommended dry docking intervals are summarised in Table 8.1. Table 8.1 Maximum periods of service for various types of antifouling product Type of formulation Maximum period of service conventional contact leaching TBT free SPC 18 months 24 months 5 years TBT SPC 5 years

It should be noted that the maximum protection periods presented in Table 8.1 are a generalisation of maximum protection periods that may be achieved within these very broad groupings. In reality, these agreed intervals reflect a compromise position reached between 100

CEPE members. In addition Table 8.1 does not provide an indication of the level of performance that can be obtained by a product specified within those time periods, or the level of performance required by a particular specification. Performance ratings are heavily dependent upon the particular coating being applied to specification (surface preparation, primers, undercoatings, dry film thickness etc.), trading and sailing pattern of the vessel and a wide variety of environmental factors.

8.3 CURRENTLY APPROVED ANTIFOULING PRODUCTS CONTAINING CHLOROTHALONIL

There are currently (November 1999) four approved antifouling products which contain chlorothalonil. These four product approvals are held by two approval holders. The distribution of chlorothalonil antifouling products by active ingredient is shown in Table 8.2. Table 8.2: The distribution of chlorothalonil antifouling products by active ingredient ACTIVE INGREDIENT (S) Chlorothalonil/Cu2O/Diuron Chlorothalonil/Cu2O/Diuron/Irgarol 1051 Chlorothalonil/Cu2O/RH-287 No 2 1 1

Information on the coating type has been received for all 4 products. In all cases the coating type that best describes the products is conventional. Both approval holders have submitted information on maximum dry-docking intervals for all of their products. This information is presented in Table 8.3, in addition to the concentration range of biocides in chlorothalonil products. Table 8.3 Range of biocides, coating type and recommended dry-docking intervals of antifouling products containing chlorothalonil
Coating type Cu2O Biocides (and range as appropriate) % w/w Chlorothalonil RH-287 No. of products having maximum recommended dry-docking intervals Diuron (36 months)

TBT-free SPC

39.1 - 46

2-3

0 - 0.9

0 - 3.7

4 products

Table 8.3 shows that the level of chlorothalonil does not exceed 3 %. All products have recommended dry docking periods within CEPE agreed periods (see Table 8.1). All four products are marketed for use on both deep sea and yachts/recreational craft. The term 'yacht' refers to those products intended for use on yachts and other recreational craft. Such products are usually re-applied on a seasonal basis (generally once a year).

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8.4 EFFICACY DATA SUBMITTED IN SUPPORT OF THE REVIEW OF THE USE OF CHLOROTHALONIL IN ANTIFOULING PRODUCTS
To date, no efficacy data have been submitted by either the manufacturers of chlorothalonil, or current approval holders. Some limited data have been identified following an extensive literature search. The data obtained are efficacy test reports. These data are included in Section 8.4.1 (laboratory [in-vitro] toxicity screening tests) only. These data have been evaluated by the HSE to assess the innate activity of the active ingredient as a potential antifouling biocide. 8.4.1 LABORATORY (IN-VITRO) TOXICITY SCREENING TESTS 8.4.1.1 Fungi In a poorly reported study, the fungicidal activity of chlorothalonil was investigated. The diametral development of fungal colonies was measured daily on selective agar-substrate, to which had been added known concentrations of chlorothalonil. The culture medium was agar cork plus sterile Whatman paper. The plates were incubated at 28 oC for 15 d. Negative (blank) concurrent controls were also run. Resulting Minimum Inhibitory Concentrations (MICs) are given in Table 8.4. Table 8.4 Minimum Inhibitory Concentrations of chlorothalonil against a range of fungal species MIC (mg l-1) at 50 % inhib. at 90 % inhib. 60 750 Chaetomium globosum 1000 1000 Phanerochaete chrysosporium 10 500 Trichoderma viride 10 800 Trichoderma reesei 10 1000 Penicillium sp. 10 200 Chaetomium celluloticum 1000 1000 Alternaria sp. 10 60 Helmintosporium sp. Fungi The authors reported that most test species showed sensitivity at 10 mg l-1 chlorothalonil, and that mycelial development was "drastically" decreased. They noted, however, that to reach total inhibition, the concentration had to be "much" higher (in excess of 1000 mg l-1). It was also noted that the laboratory conditions greatly favoured microbial development (Tonetti and Dor, 1990).

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8.4.1.2 Freshwater and Marine Algae Studies In a poorly reported study, the activity of chlorothalonil or TBTO against both freshwater and marine algae was investigated. Species tested are shown in Table 8.5. Table 8.5 Species of freshwater and marine algae tested for sensitivity to chlorothalonil Type of algae Species tested

Freshwater cyanobacteria Oscillatoria acuminata (blue-green algae) Nostoc calcicala Phormidium sp. Synechoccus sp. (Syn. Anacystis) Scytonema sp. Freshwater algae Chlorella sp. Scenedesmus acuminatus Scenedesmus sp. Enteromorpha intestinalis Ectocarpus siliculosus Dunaliella sp. Eugomontia sp. Heterococcus sp. Microcoleus sp. Navicula sp. Amphora sp. Synechoccus sp.

Marine algae

Marine microalgae and cyanobacteria (blue-green algae)

For the freshwater algae, the strain culture substrates used were BG 11 medium for cyanobacteria, and Kolkwitz medium for algae. The culture conditions were 25 oC, 5000 lux light radiation intensity and agitation at 80 oscillations min-1. The test period was 15 d. Algae development was measured by reading the absorbency value of the culture medium at the Bonet-Maurey biophotometer. The results are shown in Figures 8.1 to 8.8. It should be noted that the raw data were not included in the study report and as a result, the following graphs are approximate.

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100 90 80 Absorbance (%) 70 60 50 40 30 0 1 2 3 4 5 Concentration (ppm) Chlorothalonil TBTO

Figure 8.1. Comparison of the algicidal activity of chlorothalonil and TBTO against Oscillatoria acuminata
100 90 80 Absorbance (%) 70 60 50 40 30 0 1 2 3 4 5 Concentration (ppm) Chlorothalonil TBTO

Figure 8.2. Comparison of the algicidal activity of chlorothalonil and TBTO against Phormidium sp.
100 90 80 Absorbance (%) 70 60 50 40 30 20 0 1 2 3 4 5 Concentration (ppm) Chlorothalonil TBTO

Figure 8.3. Comparison of the algicidal activity of chlorothalonil and TBTO against Nostoc calcicola 104

75 70 65 Absorbance (%) 60 55 50 45 40 0 1 2 3 4 5.0 Concentration (ppm) Chlorothalonil TBTO

Figure 8.4. Comparison of the algicidal activity of chlorothalonil and TBTO against Chlorella sp.
100 90 80 Absorbance (%) 70 60 50 40 30 0 1 2 3 4 5 Concentration (ppm) Chlorothalonil TBTO

Figure 8.5. Comparison of the algicidal activity of chlorothalonil and TBTO against Scenedesmus sp.
100 Chlorothalonil 90 Absorbance (%) 80 70 60 50 0 1 2 Concentration (ppm) 3 4 TBTO

Figure 8.6. Comparison of the algicidal activity of chlorothalonil and TBTO against Scenedesmus acuminatus 105

100 90 80 70 60 50 40 30 0 1 2 3 4 5 Concentration (ppm) Chlorothalonil TBTO Absorbance (%)

Figure 8.7. Comparison of the algicidal activity of chlorothalonil and TBTO against Scytonema sp.
100 80 Absorbance (%) 60 40 20 0 0 1 2 3 4 5 6 7 8 9 10 Concentration (ppm) Chlorothalonil TBTO

Figure 8.8. Comparison of the algicidal activity of chlorothalonil and TBTO against Synechoccus sp. For the marine algae, the cultures were maintained at ambient temperatures for 8 weeks. Tests on the marine microalgae and cyanobacteria involved incubation "in optimum conditions". Growth was assessed by determining the optical density of cultures in sea waterborne medium. Resulting MICs are given in Table 8.6.

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Table 8.6 MICs of chlorothalonil against a range of algal species Algae MIC (mg l-1) Chlorothalonil TBTO 1 1 1 0.1 0.5 1 10 10 0.1 0.1 0.1 5 5 1 2 5 5 0.5

Enteromorpha intestinalis Ectocarpus siliculosus Synechoccus sp. Microcoleus sp. Dunaliella sp. Eugomontia sp. Amphora sp. Navicula sp. Heterococcus sp.

The authors noted that chlorothalonil exhibited good activity against freshwater algae when compared to TBTO. In particular, chlorothalonil was highly effective against the marine microalgae and cyanobacteria (Tonetti and Dor, 1990). 8.4.1.3 Summary of Laboratory Screening Tests It can be concluded from the screening studies, that toxicity of chlorothalonil was demonstrated against eight species of fungi, eight species of freshwater algae, and nine species of marine algae. Chlorothalonil appeared to be generally more active against freshwater algae than TBTO. MICs for fungi and marine algae were in the range of 0.1 >1000 mg l-1 chlorothalonil. 8.4.2 SIMULATED USE TESTS 8.4.2.1 Introduction To Raft Testing Due to the accessibility of test sites (most companies have access to raft testing facilities) and ease of operation, raft testing has been widely adopted for antifouling trials. As such, they are important indicators of the antifouling capability of biocidal active substances. They are static tests conducted usually with submerged panels coated with the test formulation. These tests are generally used to demonstrate the effectiveness of an antifouling product relative to an uncoated (blank) substrate. The method is unable to evaluate complete coating systems (especially the more advanced SPC or ablative technologies) or the relative lifetime of coatings. This means that raft testing is unable to predict the actual performance in service. Raft tests can, however, simulate the use of a product on a yacht, which will spend most of its time stationary (anchored/berthed) (Unpublished, 1996). 107

No efficacy test data generated in this way have been provided by approval holders. No raft studies have been identified from the literature. 8.4.2.2 Summary of Raft Test Data No data submitted.

8.5 OVERALL SUMMARY OF EFFICACY DATA

Only limited laboratory screening data were available. It can be concluded from the screening studies, that toxicity of chlorothalonil was demonstrated against eight species of fungi, eight species of freshwater algae, and nine species of marine algae. Chlorothalonil appeared to be generally more active against freshwater algae than TBTO. MICs for fungi and marine algae were in the range of 0.1 - >1000 mg l-1 chlorothalonil.

8.6 PROPOSALS AND DATA REQUIREMENTS

8.6.1 PROPOSALS There is some evidence that chlorothalonil will act as an active ingredient in antifouling systems. However, no simulated use tests or in-service data were provided. Ministers agreed that approvals for antifouling products containing chlorothalonil be continued pending the submission of further efficacy data. 8.6.2 DATA REQUIREMENTS To support the continued use of antifouling products containing chlorothalonil, raft test or alternatively field trial/in service monitoring data are required. These data should cover: (i) The efficacy of TBT-free SPC coating antifouling formulations that contain chlorothalonil. The full nature and extent of data required is to be outlined to individual approval holders, as appropriate, by the HSE. In all cases, data should be provided by approval holders in support of the coating type(s) that best befits their products. If a product is assigned to a different and more accurate coating type by an approval holder, and is, as a consequence, not supported by existing information, appropriate efficacy data must be submitted. Alternatively for (i), justification or evidence based on data may be provided through the submission of bridging studies. If the approval holder undertakes to conduct new studies, the HSE will require evidence that such work is under way after a period of six months. In all cases, suitable efficacy data to be provided within three years.

108

OVERALL RECOMMENDATIONS AND DATA REQUIREMENTS

1. The following recommendations and data requirements were agreed by Ministers: 1. All amateur uses of antifouling products containing chlorothalonil are to be revoked because information from humans indicates that the risk of skin sensitisation is unacceptable. 2. Approvals for the professional use of antifouling products containing chlorothalonil at a maximum concentration of 5 % w/w may be continued for professional use by brush, roller and spray, subject to the data requirements and conditions detailed below: PHYSICAL CHEMISTRY For technical chlorothalonil: 1 All data to be submitted within 1 year unless otherwise specified. Approval Holders must demonstrate to HSE how they are implementing the requirements within 6 months. From Company No. 1: a) b) c) d) e) f) g) Address of manufacturing site. Results from typical batch analyses conducted on technical chlorothalonil, with associated test reports (e.g. chromatographic traces should be included). A minimum maximum range of purity for the active ingredient should also be stated. UV and IR traces to support the results provided. Further spectral data to include 13C NMR spectra and MS (traces should be provided). Surface tension including full test report. Full test reports for: melting point, boiling point, relative density, vapour pressure, solubility in water, partition coefficient, and explosive properties. Protocol reports, plus typical experimental results including example traces and validation data, for the analytical methods provided for the determination of chlorothalonil in the technical material and water. The analytical method used to determine chlorothalonil in water should be able to determine the active substance to 0.1 g l-1. Analytical methods to determine chlorothalonil in antifouling products. Protocol reports, plus typical experimental results including example traces and validation data should be included.

h)

From Company No. 2: a) b) c) d) e) BSI name, IUPAC name, other names, codes and synonyms. Identification numbers including CAS and EINECS numbers. Molecular mass, molecular formula, structural formula. Site of manufacture (including company name and address). Technical specification including min. - max. purity of active substance expressed in %w/w or g kg -1 and identification and quantification of significant impurities and those of toxicological concern expressed as max. %w/w or g kg -1. 109

f) g) h)

i)

j)

Results from typical batch analyses conducted on technical chlorothalonil, with associated test reports (e.g. chromatographic traces should be included). Spectral data, to include UV, IR, 13C NMR spectra and MS (traces should be provided). The following endpoints (including full test reports): melting point, boiling point, relative density, surface tension, vapour pressure, solubility in water, partition coefficient log Pow, flammability*, oxidising* and explosive properties*. * As an alternative, reasoned cases may be provided. Analytical methods for determination of chlorothalonil and impurities in technical material to support the technical specification data requirement in e). Protocol reports, plus typical experimental results including example traces and validation data should be included. Analytical method to determine chlorothalonil in water and in antifouling products formulations. The analytical method used to determine chlorothalonil in water should be able to determine the active substance to 0.1 g l-1.

From all Approval Holders Storage stability studies on representative current products, under ambient conditions in the product packaging. If the approval holder undertakes to conduct new studies, HSE will require evidence that such work is underway after a period of 6 months. New long term storage stability studies should be provided within 3 years. Protocols are to be agreed with HSE. 1. 2. HUMAN HEALTH Data are to be submitted within 1 year. Approval holders must demonstrate to HSE how they are implementing the requirements within 6 months. a) A study of the dermal penetration of chlorothalonil from formulations representative of approved antifouling products, to be submitted within 1 year. This study may be carried out in vitro or in vivo. The protocol should be discussed with HSE prior to commencement. Professional operators (sprayers) exposed to antifouling products containing chlorothalonil must wear RPE. Appropriate RPE includes air-fed respiratory equipment with combined protective helmet and visor to protect the skin of the head and neck. Impairment of vision should be avoided. For workers other than sprayers in the vicinity of the spray plume, RPE of an equivalent standard to FFP3 should be worn. For other workers, the need for RPE should be informed by a COSHH assessment. Unprotected persons should be kept out of treatment areas to minimise the risks posed by exposure to this compound. All professional operators exposed to antifouling products containing chlorothalonil should wear a disposable coverall with hood (providing head protection) and a second overall beneath this coverall of a contrasting colour to the antifouling product being applied. All bare skin should be covered. The disposable coverall should normally be

b)

c)

d)

110

used for no more than one spraying session. The second overall should be changed regularly and whenever product break-through has been detected. e) Professional operators working with antifouling products containing chlorothalonil should wear impermeable gloves of a type recommended by the antifouling manufacturer as suitable for use with the formulation. These gloves should be changed regularly, e.g. after one or two days use. Operators should wear impermeable (and non-slip) footwear that protects the lower leg. The following approval conditions are to appear on products' Notice of Approval and Schedules and they should be reflected on product labels using the following precautionary phrases: WEAR SUITABLE PROTECTIVE CLOTHING (COVERALLS OF A CONTRASTING COLOUR TO THE PRODUCT BEING APPLIED, BENEATH A DISPOSABLE COVERALL WITH HOOD), SUITABLE GLOVES, AND IMPERVIOUS FOOTWEAR THAT PROTECTS THE LOWER LEG. DISPOSE OF PROTECTIVE GLOVES AFTER USE If the product is to be applied by spray: UNPROTECTED PERSONS SHOULD BE KEPT AWAY FROM TREATMENT AREAS DO NOT BREATHE SPRAY MIST WEAR SUITABLE RESPIRATORY EQUIPMENT SUCH AS AIR-FED RESPIRATORY EQUIPMENT WITH COMBINED PROTECTIVE HELMET AND VISOR WHEN SPRAYING. WEAR SUITABLE RESPIRATORY EQUIPMENT (SUCH AS FFP3 OR AN EQUIVALENT STANDARD) WHEN WORKING IN THE VICINITY OF THE SPRAY ENVIRONMENT The following data requirements are to be submitted within a total of 3 years, with shorter timetables for some requirements as outlined. Approval holders must demonstrate to HSE how they are implementing the data requirements within 6 months. a) Inhibition of microbial respiration in sewage sludge (core requirement) to be submitted within 12 months.

f)

b) Fish early-life stage test (sheepshead minnow preferable species) to be submitted within 12 months. c) Clarification that the leaching rate of the parent ai used for the risk assessment is representative of current products. This information is to be submitted within 12 111

months. (Repeated REMA calculations will be carried out by HSE with any new data supplied) d) Should the leaching data submitted as part of requirement c) fail to reduce the predicted environmental concentrations of chlorothalonil to acceptable levels (assuming 100 % use on pleasure vessels) a monitoring programme will be necessary. In the first instance, HSE should liaise with DETR, EA and English Nature and investigate the potential for further monitoring via the National Marine Monitoring Programme or other organisations. e) Confirmation that chlorothalonil significantly degrades in marine sediment-water systems, and identification of any subsequent persistent metabolites to be submitted within 12 months. Once the significance and persistence of the metabolites have been established additional ecotoxicology, fate and behaviour data may be required in order to permit a risk assessment to the same level as that of the parent compound. The specific data required should be discussed with HSE on completion of data requirement e) and all subsequent data requirements submitted within a further 2 years.

f)

g) A study to address bioaccumulation of chlorothalonil in shellfish, in order to address the potential risks to consumers via the food chain, should be submitted within 2 years. EFFICACY The following efficacy data are to be provided within 3 years. If approval holders undertake to conduct new studies, they must demonstrate to HSE how they are implementing the data requirements within 6 months. To support the continued use of antifouling products containing chlorothalonil, raft test or alternatively field trial/in service monitoring data are required. These data should cover: a) The efficacy of TBT-free SPC coating antifouling formulations that contain chlorothalonil.

The full nature and extent of data required is to be outlined to individual approval holders, as appropriate, by the HSE. In all cases, data should be provided by approval holders in support of the coating type(s) that best befits their products. If a product is assigned to a different and more accurate coating type by an approval holder, and is, as a consequence, not supported by existing information, appropriate efficacy data must be submitted. Alternatively for a), justification or evidence based on data may be provided through the submission of bridging studies.

112

REFERENCES
PHYSICAL CHEMISTRY (CHAPTER 2)
ACP (1994). Chlorothalonil: risk to operators and consumers. ACP published evaluation No. 112. ACP Secretariat, PSD, Mallard House, Kings Pool, 3 Peasholme Green, York, YO1 7PX. Unpublished (1999). Physical chemistry data.

MAMMALIAN TOXICOLOGY AND RISK ASSESSMENT (CHAPTERS 3 AND 4)

ACP (1994). Chlorothalonil: risk to operators and consumers. ACP published evaluation No. 112. ACP Secretariat, PSD, Mallard House, Kings Pool, 3 Peasholme Green, York, YO1 7PX. Bach B and Pedersen NB (1980). Contact dermatitis from a wood preservative containing tetrachloroisophthalonitrile. Contact Dermatitis 6, 142. Bruynzeel DP and van Ketel WG (1986). Contact dermatitis due to chlorothalonil in floriculture. Contact Dermatitis 14, 67 - 68. Dannaker CJ, Maibach HI and OMalley M (1993). Contact urticara and anaphylaxis to the fungicide chlorothalonil. Cutis 52, 312 - 315. Dolara P, Torricelli F and Antonelli N (1994). Cytogenetic effects on human lymphocytes of a mixture of fifteen pesticides commonly used in Italy. Mutation Research 325, 47-51. Environment Agency (1998). Survey of Manufacturers, Chandlers (Suppliers) and treatment Sites. Environment Agency Research and Development Technical Report P215. WRc plc ISBN 1 873160 74 7). Galloway SM, Armstrong MJ, Reuben C et al (1987). Chromosome aberrations and sister chromatid exchanges in Chinese hamster ovary cells: evaluations of 108 chemicals. Environ Mol Mutagen 10, Suppl 10, 1-175. Garrod ANI, Guiver R and Rimmer DA (2000). Potential exposure of amateurs (consumers) through painting wood preservative antifoulant preparations. Ann Occ Hyg 44(6), 421-426. Hillenweck A, Corpet DE, Killeen JC Jr, et al (1998). Ex vivo gastrointestinal biotransformation of chlorothalonil in the germ-free and conventional rat. Xenobiotica 28, 1017-1028. HSE (2000). Dermal exposure to non-agricultural pesticides. Exposure assessment document (EH 74/3). HSE Books, Sudbury, UK. ISBN 0-7176-1718-1.

113

Huang J, Aoyama K, Ueda A et al (1995). Respiratory effects and skin allergy in workers exposed to tetrachloroisophthalonitrile. Bull Environ Contam Toxicol 55, 320 - 324. IOM (1994). Occupational hygiene assessment of exposure to insecticides and the effectiveness of protective clothing during sheep dipping operations. IOM Report No. TM/94/04. Johnsson M, Buhagen M, Liera HL et al (1983). Fungicide-induced contact dermatitis. Contact Dermatitis 9, 285 - 288. Lindn C (1990). Facial dermatitis caused by chlorothalonil in a paint. Contact Dermatitis 22, 206 - 211. Lebailly P, Vigreux C, Godard T et al (1997). Assessment of DNA damage induced in vitro by etoposide and two fungicides (carbendazim and chlorothalonil) in human lymphocytes with the comet assay. Mutation Research 375, 205-217. Lodovici M, Casalini C, Briani C et al (1997). Oxidative liver DNA damage in rats treated with pesticide mixtures. Toxicology 117, 55-60. Matsushita T and Aoyama K (1981). Cross reactions between some pesticides and the fungicide Benomyl in contact allergy. Industrial Health 19, 77-83. Matsushita S, Kanekura T, Saruwatari K et al (1996). Photoallergic contact dermatitis due to Daconil. Contact Dermatitis 35, 115 - 116. McGregor DB, Brown A, Cattanach P et al (1988). Responses of the L5178Y tk+/tk- mouse lymphoma cell forward mutation assay: III 72 coded chemicals. Environ Mol Mutagen 12, 85-154. Meding B (1986). Contact dermatitis from tetrachloroisophthalonitrile in paint. Contact Dermatitis 15, 187. Mizens M, Killeen JC Jr and Eilrich GL (1998). The mutagenic potential of chlorothalonil: in vivo chromosome aberration studies. Mutation Research 403, 269-272. OMalley MO, Rodriguez P and Maibach HI (1995). Pesticide patch testing: California nursery workers and controls. Contact Dermatitis 32, 61 - 63. Penagos H, Jimenez V, Fallas V, et al (1996) Chlorothalonil, a possible cause of erythema dyschromicum perstans (ashy dermatitis). Contact Dermatitis 35, 214 - 218. Shah RG, Lagueux J, Kapur S et al (1997). Determination of genotoxicity of the metabolites of the pesticides Guthion, Sencor, Lorox, Reglone, Daconil and Admire by 32Ppostlabelling. Molecular and Cellular Biochemistry 169, 177-184. Ueda A, Aoyama K, Manda F, et al (1994). Delayed-type allergenicity of triflorine (Saprol). Contact Dermatitis 31, 140 - 145. Unpublished, (1985). Bravo, Daconil 2787, Broad Spectrum Fungicide. 114

Unpublished, (1987). Acute oral toxicity of Flexgard III in Sprague-Dawley rats. Unpublished, (1987a). Acute dermal toxicity of Flexgard III in Sprague-Dawley rats. Unpublished, (1987b). Primary dermal irritation of Flexgard III in New Zealand white rabbits. Unpublished, (1987c). Primary eye irritation of Flexgard III in New Zealand white rabbits. Unpublished, (1988). Dermal sensitization study (closed patch repeated insult) in guinea pigs with Demidekk Neutral Base A, Demidekk Neutral Base B and Demidekk Neutral Base C. Unpublished, (1995). Potential exposure of workers to chlorothalonil when handling and applying paint containing chlorothalonil. Unpublished, (1996). A comparison of two different methods for assessment of dermal exposure to non-agricultural pesticides in three sectors. Unpublished, (1998). Measurement of potential dermal exposure in Greece and Spain with patch and whole body dosimetry techniques. Vigreux C, Poul JM, Deslandes E et al (1998). DNA damaging effects of pesticides measured by the single cell gel electrophoresis assay (comet assay) and the chromosomal aberration test, in CHOK1 cells. Mutation Research 419, 79-90. Yamano T and Morita S (1995). Effects of pesticides on isolated rat hepatocytes, mitochondria, and microsomes II. Arch Environ Contam Toxicol 28, 1-7.

ENVIRONMENTAL DATA (CHAPTERS 5, 6 AND 7)

ACP (2002): Environmental Risk Assessment of Booster Biocides in Antifouling Products. ACP Evaluation No. 205. ACP Secretariat, PSD, Mallard House, Kings Pool, 3 Peasholme Green, York YO1 7PX. Callow ME and Willingham GL (1996). Degradation of antifouling biocides. Biofouling 10 (1/3), 239-49. Caux P-Y, Kent RA, Fan GT and Stephenson GL (1996). Environmental fate and effects of chlorothalonil: A Canadian perspective. Crit Rev Environ Sci Technol 26 (1), 45-93. Davies PE (1987). Physiological anatomic and behavioural changes in the respiratory system of Salmo-gairdneri Rich. on acute and chronic exposure to chlorothalonil. Comp Biochem Physiol C Comp Pharmacol Toxicol 88 (1), 113-120. Davies PE (1988). Disappearance rates of chlorothalonil in the aquatic environment. Bull Environ Contam Toxicol 40 (3), 405 (5).

115

Davies PE, Cook LSJ and Goenarso D (1994). Sub lethal responses to pesticides of several species of Australian freshwater fish and crustaceans and rainbow trout. Env Tox & Chem 13 (8),1341-1354. EURATGD (1996). Technical Guidance Document in Support of Commission Directive 93/97/EEC on Risk Assessment for New Notified Substances and Commission Regulation (EC) No 1488/94 on Risk Assessment for Existing Substances. Part II. ECSC-EC-EAEC, Brussels. Luxembourg. Gajbhiye VT, Jain HK and Agnihotri NP (1989). Persistence of chlorothalonil in soil water and sediment. Pesticides (Bombay) 23 (9), 32J-32L. Gallagher EP, Cattley RC and Di Giulio RT (1992a). The acute toxicity and sub-lethal effects of chlorothalonil in channel catfish Ictalurus-punctatus. Chemosphere 24 (1), 3-10. Gallagher EP, Canada AT and Di Giulo RT (1992b). The protective role of glutathione in chlorothalonil-induced toxicity to channel catfish. Aquat Toxicol 23 (3-4), 155 (14). Tsuda T, Aoki S, Kojima M and Fujita T (1992). Accumulation and excretion of pesticides used in golf courses by carp (Cyprinus carpio) and willow shiner (Gnathopogon caerulescens). Comparative biochemistry and physiology: C: Comparative pharmacology and toxicology. 101 (1), 63-66. Unpublished (1972). Exposure of fish to distribution and elimination of residues.
14

C-labelled technical DAC-2787: Accumulation,

Unpublished (1976). Hydrolysis of Daconil and its metabolite, 4-hydroxy-25,6trichloroisophthalonitrile, in the absence of light at pH levels of 5, 7 and 9. Unpublished (1977). Acute toxicity of DS-2787 (technical chlorothalonil) to the water flea (Daphnia magna). Unpublished (1980a). Effect of 2,4,5,6,-Tetrachlorisophthalonitrile (chlorothalonil, DS-2787) upon the microbial utilization of selected macromolecules in soil. Unpublished (1980b). Accumulation, distribution and depuration of DS-2787 in bluegill sunfish under flow-through conditions. Unpublished (1981a). Effect of 2,4,5,6,-Tetrachlorisophthalonitrile (chlorothalonil, DS-2787) upon non-symbiotic nitrogen fixing soil micro-organisms. Unpublished (1981b). Effect of 2,4,5,6,-Tetrachlorisophthalonitrile (chlorothalonil, DS-2787) upon nitrogen transformation in soil. Unpublished (1981c). Characterisation and quantitation of 14C-residues in water and fish exposed to DS-2787 in a flow-through aquatic system. Unpublished (1981d). Accumulation, distribution and depuration of DS-2787 in channel catfish under static conditions.

116

Unpublished (1981e). Characterisation and quantitation of 14C-residues in water, soil and fish exposed to 14C-chlorothalonil, DS-2787 in a static aquatic system. Unpublished (1982). Adsorption and Desorption of chlorothalonil to soils. Unpublished (1990). Leach rate determinations of antifoulant coatings/flexigard III containing TCIN. Unpublished (1992a). Bravo-720 Acute toxicity to the daphnids (Daphnia magna). Unpublished (1992b). Bravo-720 Acute toxicity to bluegill sunfish (Lepomis macrochirus) under flow-through conditions. Unpublished (1992c). Acute toxicity to rainbow trout (Oncorhynchus mykiss) under flowthrough conditions with Bravo-720. Unpublished (1995a). Yacht antifouling coating system release rates of chlorothalonil and copper in seawater. Unpublished (1995b). Summary data prepared in support of the European Plant Protection Products Directive (PPPD) review of chlorothalonil as an agricultural fungicide. Unpublished (1998). Pers Comm. Walker WW, Cripe CR and Prichard PH (1988). Biological and abiotic degradation of xenobiotic compounds in in-vitro estuarine water and sediment/water systems. Chemosphere 17 (12), 2255 (16).

EFFICACY (CHAPTER 8)
Callow M (1990). Ship Fouling: Problems and solutions. Chemistry & Industry 5, 123-127, 5th March 1990. Clare AS, Rittschof D, Gerhart DJ and Maki JS (1992). Molecular approaches to nontoxic antifouling. Invertebrate Reproduction and Development 22:1-3, 67-76. Hunter JE (1994). Biocides Directive Steering Group - Antifouling coatings. Paper presented to CEPE Antifouling Manufacturers Working Group. Tonetti P and Dor F (1990). A new biocide in the formulation of marine paints. Pitture Vernici 1990, 66 (46), 5-13. Unpublished (1996). Pers Comm. US Naval Institute, 1981. "Marine Fouling and its Prevention", Annapolis: Woods Hole Oceanographic Institute, US Naval Institute, 165-207, (1981).

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APPENDIX 1
CALCULATIONS FOR EXPOSURE ASSESSMENT
The following database models were initially presented in guidance document EH74/3 (HSE, 2000). Defaults assumed - spray applications: operator weight 60 kg operator respired volume 1.25 m3 h-1 median shift length 3h Table 1.1 summary of exposure data for sprayers (mg h-1 in-use product)
central tendency 6170 6170 4.5% 4% 60 60 6.6 mg m-3 6 mg m-3

frequency potential dermal 100% weighted indicative value clothing penetration 93% weighted indicative value in-glove exposure 100% weighted indicative value inhalation exposure 91% weighted indicative value

95th% or * worst case 44700

241 64.6 mg m-3

Table 1.2

summary of exposure data for pot-men (mg h-1 in-use product)

central tendency 2940 2940 7% 4% 34.9 35 0.9 mg m-3 0.6 mg m-3

frequency potential dermal 100% weighted indicative value clothing penetration 59% weighted indicative value in-glove exposure 100% weighted indicative value inhalation exposure 68% weighted indicative value

95th% or * worst case 15000

1380 * 42 mg m-3 *

Table 1.3 product)

summary of exposure data for ancillary workers (mg h-1 in-use

central tendency 885 885 7% 4% 35 35 1.7 mg m-3 95th% or * worst case 44700

frequency potential dermal 100% weighted indicative value clothing penetration 59% weighted indicative value in-glove exposure 100% weighted indicative value inhalation exposure 50%

241 * 4.8 mg m-3 *

118

weighted indicative value

0.8 mg m-3

The original reports from which these tables data are derived are available for scrutiny at HSE Bootle. The number of exposure data and the exposure ranges are in Table 1.4 Table 1.4
exposure potential dermal mg h-1 product in-gloves mg h-1 product inhaled mg m-3 product no of data 29 19 20

Summary of exposure data and ranges - antifoulant spraying

sprayer range 52.2 - 74100 0.18 - 252 0.04 - 79.4 pot-men and other operators no of data range 28 16.2 - 18200 17 16 0.31 - 1380 0.04 - 41.6

Table 1.5

Exposure estimate (product) - sprayer

central tendency worst case 44700 3 134000 4 5360 241 3 723 6080 64.6 3 3.75 242 4.84 6170 3 18510 4 740 60 3 180 920 6 3 3.75 22.5 0.45

Coveralls potential dermal exposure, mg h-1 work time per day, h daily deposit on clothes penetration, % dermal exposure to product, mg Gloves dermal exposure inside, mg h-1 work time per day dermal exposure to product, mg Total dermal exposure antifoulant product, mg Inhaled concentration, mg m-3 work time per day, h inhaled air volume, m3 inhaled product, mg - no RPE inhaled product, mg - RPE x PF 50 PF - RPE protection factor

119

Table 1.6

Exposure estimate (product) - pot-man

central tendency worst case 15000 3 45000 4 1800 1380 3 4140 5940 42 3 3.75 158 2940 3 8820 4 353 35 3 105 458 0.6 3 3.75 2.25

Coveralls potential dermal exposure, mg h-1 work time per day, h daily deposit on clothes penetration, % dermal exposure to product, mg Gloves dermal exposure inside, mg h-1 work time per day dermal exposure to product, mg Total dermal exposure antifoulant product, mg Inhaled concentration, mg m-3 work time per day, h inhaled air volume, m3 inhaled product, mg - no RPE

Table 1.7

Exposure estimate (product) - ancillary workers

central tendency worst case 3470 3 10400 4 416 180 3 540 956 4.8 3 3.75 18 885 3 2660 4 106 35 3 105 211 0.8 3 3.75 3

Coveralls potential dermal exposure, mg h-1 work time per day, h daily deposit on clothes penetration, % dermal exposure to product, mg Gloves dermal exposure inside, mg h-1 work time per day dermal exposure to product, mg Total dermal exposure antifoulant product, mg Inhaled concentration, mg m-3 work time per day, h inhaled air volume, m3 inhaled product, mg - no RPE

120

Defaults assumed - brush and roller applications: user weight 60 kg operator respired volume 1.25 m3 h-1 median job duration 1.5 h professional chandlers undertake the same tasks as amateur users of antifoulants Table 1.8 summary of exposure data for amateurs (mg h-1 in-use product)
central tendency 1020 1020 42% 5% ** 31.2 31 0.04 mg m-3 0.02 mg m-3 frequency potential dermal 100% weighted indicative value clothing penetration 11% weighted indicative value in-glove exposure 100% weighted indicative value bare hand exposure 100% inhalation exposure 44% weighted indicative value 95th% or * worst case 6480 *

1110 * 4400 * 0.11 mg m-3 *

** It is possible that amateurs wear only minimal clothing when applying antifoulant products. A weighted indicative value for penetration in such cases is a default 50%. Table 1.9 Exposure estimate (product) - amateur user
central tendency Coveralls potential dermal exposure, mg h-1 work time per day, h daily deposit on clothes penetration, % dermal exposure to product, mg Gloves dermal exposure inside, mg h-1 work time per day dermal exposure to product, mg Total dermal exposure antifoulant product, mg Inhaled concentration, mg m-3 work time per day, h inhaled air volume, m3 inhaled product, mg - no RPE 1020 1.5 1530 5 76.5 31 1.5 46.5 123 0.02 1.5 1.88 0.04 worst case 6480 1.5 9720 50 ** 4860 4400 ** 1.5 6660 11500 0.11 1.5 1.88 0.21

** worst case - no gloves, minimal clothing.

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Table 1.10

Exposure estimate (product) - professional chandler

central tendency worst case 6480 1.5 9720 5 486 1110 1.5 1670 2160 0.11 1.5 1.88 0.21 1020 1.5 1530 5 76.5 31 1.5 46.5 123 0.02 1.5 1.88 0.04

Coveralls potential dermal exposure, mg h-1 work time per day, h daily deposit on clothes penetration, % dermal exposure to product, mg Gloves dermal exposure inside, mg h-1 work time per day dermal exposure to product, mg Total dermal exposure antifoulant product, mg Inhaled concentration, mg m-3 work time per day, h inhaled air volume, m3 inhaled product, mg - no RPE

Table 1.11 Sample Type Inside PPE On head Inside gloves Outside gloves Inhaled (mg m-3) Job (min)

Industry data (mg product min-1 unless otherwise stated) Water-based product sprayed indoors Range Median 1.76 4.25 15.5 3.04 7.98 13.4 0.1 0.19 0.52 14.2 36 63.6 67 - 194 126 22 - 44 33 Water-based product sprayed outdoors Range Median 0.34 0.84 2.6 1.06 2.5 8.36 0.02 0.04 0.09 12 - 40.8 29.6 6.1 - 30 35 - 81 14 49 Solvent-based product sprayed outdoors Range Median 0.54 1.05 1.61 1.37 3.72 12.3 0.02 0.04 0.2 17.1 - 60 30.4 5.7 - 18 35 - 53 11 38 All data Median 1.15 3.8 0.6 30.7 17 38

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APPENDIX 2
CURRENT ANTIFOULING COATING TYPES

Coating type

Description and Properties

Soluble matrix In coatings of this type, the biocide(s) have been physically mixed ('freely (conventional) associated') into a rosin matrix. Upon exposure to sea water, the slightly

acidic matrix slowly dissolves releasing the biocide(s) into the water (sea water is slightly alkaline (pH 8) and the acidic matrix readily dissolves). Continuous dissolution of the coating surface occurs, resulting in fresh biocide(s) being released until eventually the film is exhausted. The soluble matrix coatings have poor mechanical properties which limit film thickness and hence the coating lifetime attainable to approximately 12-18 months. As the matrix rosin is a natural product, batches differ and therefore coating lifetime is unpredictable.
Insoluble matrix (contact leaching or long life)

Within this type of coating, the binder or matrix is insoluble, and the biocide(s) is physically mixed into the matrix (often at higher concentrations than is the case with the conventional coatings). As sea water enters the paint film the biocides are released by dissolution and diffusion from within the insoluble matrix. This type of coating has a high initial release rate, which decreases exponentially with time as the biocide(s) has further to travel through the paint film. This release process continues until exhaustion of the coating. The higher mechanical strength obtained with these coatings allow applications of thicker systems, and as a consequence, coating lifetimes of approximately 24 months are attainable. In this type of coating the TBT biocide is chemically bound to the binder of the paint, a methacrylic acid/methylmethacrylate copolymer matrix into which other biocides can be incorporated. The copolymer hydrolyses at a predictable rate in sea water (depending on temperature, pH and rate of movement of a vessel through water), releasing the biocide(s) into the surrounding water, and creating a localised concentration at the paint surface discouraging the growth of settling organisms. This hydrolysis results in a softening of the surface layer of the copolymer and, together with the physical wearing away of the binder by the action of passing sea water (polishing), exposes fresh surface layers. This mode of action with biocide release and polishing rate are both dependent on the same (chemical) process. The paint film thus smoothes, reducing drag and turbulence until eventually, through these processes, the whole of the coating is exhausted. After initial rapid release, a steady biocide release is achieved; the life of the coating is proportional to its thickness and is accurately predictable.

TBT self polishing copolymer coatings

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Coating type (continued)

Description and Properties (continued)

The copolymer has a high mechanical strength, allowing build up of very thick systems and hence correspondingly long coating lifetimes, typically up to 5 years.

TBT-free self Coatings of this type rely on soluble medium, such as rosin, in combination polishing with insoluble polymers to form a matrix which wears away physically at a copolymer controlled rate. The biocide(s) is mixed into the matrix and released by coatings dissolution at a rate determined by the rate of physical ablation of the

polymer. The physical ablation process is less controlled and predictable than the chemical ablation process. Therefore the steady release rate, predictable life, smoothing and recoating properties of the TBT copolymer coatings are difficult to achieve with this group of coatings. These tin-free copolymer coatings have, to date, demonstrated that dry docking intervals of 5 years can be achieved by the better performers within this group of products.

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