ESTABLISHMENT OF REENTRY INTERVALS FOR

ORGANOPHOSPHATE-TREATED COTTON FIELDS

BASED ON HUMAN DATA:
III. 12 TO 72 HOURS POST-TREATMENT
EXPOSURE TO MONOCROTOPHOS,
ETHYL- AND METHYL PARATHION 1
G. W. WARE, D. P. MORGAN,
BETTY J. ESTESEN, and W. P. CAHILL
Department of" Entomology, University o f Arizona
7~cson, Arizona 85721

Five human volunteers entered methyl parathion, ethyl parathion, or mono-

crotophos treated cotton fields for five-hr exposure periods when the residues of
the respective pesticides had aged 12 hr, 24 and 48 hr, and 48 and 72 hr. Foliage
residues of methyl parathion disappeared fastest, those of monocrotophos slowest.
Personal exposure to pesticide was evaluated from contamination of skin, clothing,
and ambient air, while actual absorption of chemical was assessed from pesticide
concentration in blood,, urinary metabolite excretion, and effects on blood
cholinesterase activities. There was good correspondence between magnitudes of
foliar residue, estimates of personal contamination, and measures of chemical ab-
sorption. Field exposures caused no symptoms or clinical signs of organophosphate
poisoning and depressed averaged blood cholinesterase activities by no more than
14% of pre-exposure levels.

This is an extension of information derived from studies conducted in 1971-72 bearing

on the potential adverse effects of monocrotophos (Azodrin), methyl- and ethyl parathion
(Ware e t aL 1973, Ware e t aL 1974a). The first study utilized 30-min human exposures 0,
12, 24, 48, and 72-hr post-treatment, while the second involved five-hr exposures to
cotton fields 24 hr after treatment. In the latter study there was no effect on cholin-
esterase (ChE) from exposure to methyl parathion; however, monocrotophos produced a
drop in red blood cell cholinesterase (RBC ChE) while ethyl parathion resulted in a slight
lowering of both RBC and plasma pseudocholinesterase activity (plasma ChE). Dermal
penetration was the principal route of insecticide absorption in that work situation.

The purpose of these studies was again to test empirically the safety of reentering cot-
ton fields 12 to 72 hr following application of three organophosphate insecticides com-
monly used in cotton culture. For five continuous hours, five volunteers carried on the

1Contribution to Regional Project W-45, "Residues of Selected Pesticides and Related Chemicalsin the
Agricultural Environment-Their Nature, Distribution, Persistence, and Toxicological Implications.'"
University of Arizona Agricultural Experiment Station #2281.

Archives of Environmental Contamination 289

and Toxicology Vol. 3,289-306 (1975)
9 1975 by Springer-Verlag New York Inc.
290 G . W . Ware et al.

duties involved in inspection of cotton plants for evidence of insect damage while the
following parameters relevant to assessment of insecticide exposure hazard were measured:

1. Foliar insecticide residues at varying levels of canopy during field exposure.

2. Insecticide contamination of hands, upper arms, torso and legs.
3. Adsorption of insecticide onto clothing.
4. Ambient air concentrations of insecticide.
5. Insecticide present in the blood.
6. Urinary excretion of the metabolite, p-nitrophenol (PNP).
7. Effect of field exposures on plasma and red blood cell CHE activities.

Experimental
Fourteen volunteers were involved in the following five reentry experiments, five
participating in each reentry. In all experiments the clothing, exposure times, field
activities, biomedical, field and laboratory procedures were those described by Ware et al.
(1974a), and followed the paired reentry protocol presented in Table I. This protocol re-
quired 300 min of movement through a treated field. Precise times of activities were
variable depending on the time used in activities out of the field. Consequently from 6.5
to 7 hr passed while acquiring five hr of actual field exposure.

Methyl parathion. Methyl parathion (phosphorothioic acid, O,O-dimethyl O-p-nitro-

phenyl ester) was applied to short staple (upland) cotton at the recommended rate of
1.0 lb M/acre as an emulsion in nine gal of spray per acre with an International 12-row,
high-clearance (Hi-Boy) self-propelled ground sprayer, using three #6 cone nozzles per
row and 40 psi beginning at 6:15 p.m. on September 4, 1973. The field was planted in
40-in. rows, averaged approximately 5.5 ft in height, varying from 4.0 to 6.5 ft, and had
abundant squares, blooms and bolls, with foliage from adjoining rows intertwined. This
resulted in maximum contact of personnel with foliage so that jeans were stained green
at the end of the study.

The spray was applied to a 12-acre block, 168 rows wide by 1000 ft long in a field near
Coolidge, Arizona. This field had been treated twice previously on July 27 and August 9
with methyl parathion at 1.0 lb M/acre.

The temperature at time of application was 960F, reaching a minimum of 76~ for
the 12-hr residue aging period.

Monoerotophos. Monocrotophos (phosphoric acid, dimethyl ester, ester with cis-3-

hydroxy-N-methyl crotonamide) was applied on August 13 at the rate of 1.0 lb M/acre
to a 10-acre block in the same field as "the first with the same ground equipment and
application parameters.

The temperature at time of application was 82~ with a maximum of 1030F and a
minimum of 82~ for the 48-hr, and a maximum of 105~ and a minimum of 81~ for
the 72-hr residue aging periods.
 Establishment of Reentry Intervals for Cotton Fields 291

Table I. Protocol for 24 to 48 hour paired reentries into pesticide-treated cotton fields

Day prior to 1st field reentry

7:00-10:00 A.M. Field treated with insecticide
Leaves sampled and extracted for initial
insecticide residue
24-hr control urine begun (1)a
Blood sampled for ChE and insecticide (1)

Day of 1st field reentry

Before reentry Dressed in field clothing, air samplers activated,
bladder emptied, 24-hr control urine (1)
completed.

7:00 A.M. Crew 1 entered field

Leaves sampled and extracted for 1st reentry
residue

7:00-10:00 A.M. Cotton inspected (1)

24-hr control urine begun (2)
Blood sampled for ChE and insecticide (2)

10:00 A.M. Blood sampled for ChE and insecticide (1)

Air sampler glycol exchanged (1)

10:30-11:30 A.M. Cotton inspected (1)

11:30 A.M. Hands and arms washed with solvent (1)

11:30-12:30 P.M. Lunch (1)

12:30-2:30 P.M. Cotton inspected (1)

2:30 P.M. Blood sampled for ChE and insecticide (1)

Hands and arms washed with solvent
Torsos and legs swabbed with ethanol-pads
Air sampler glycols stored
Clothing removed and bagged

Day of 2nd field reentry

Before reentry Dressed in field clothing, air samplers activated,
bladder emptied, 24-hr control urine
completed (2)

7:00 A.M. Crew 2 entered field

Leaves sampled and extracted for 2nd reentry
residue

7:00-10:00 A.M. Cotton inspected (2)

8:00 A.M. 2nd 24-hr urine completed (1)

Blood sampled for ChE and insecticide (1)
Continued
 292 G. W. Ware et al.

Table I. C o n t i n u e d

10:00 A.M. 2:30 P.M. Procedures for Crew 2 identical to 1st reentry

Day following 2nd field reentry

8:00 A.M. 3rd 24-hr urine completed (1)
2nd 24-hr urine completed (2)
Blood sampled for ChE and insecticide (2)

Second day following 2nd reentry

8:00 A.M. 3rd 24-hr urine completed (2)

aDenotes Reentry Crew 1 or 2 activities.

Ethyl parathion. Ethyl parathion (phosphorothioic acid, O,O-diethyl O-p-nitrophenyl

ester) was applied on August 20 at the rate of 1.0 lb M/acre to a 10-acre block of cotton,
which averaged about four ft in height, with the same ground equipment and application
parameters as previously described, but in a different field. Previous application of insec-
ticide to the field was on July 4 and July 27, when toxaphene-DDT, 2 + 1 lbs AI/acre,
were applied by ground equipment as part of a large scale experiment.

The temperature at time of application was 80~ with a maximum of 99~ and a
minimum of 78~ for the 24-hr and a maximum of 101~ and a minimum of 78~ for
the 48-hr residue aging period.2

Residue sampling. Leaf samples were collected immediately following treatment of the
field and at 12, 24, 48, and 72 hr, depending on the reentry schedule. Three samples con-
sisted of 50 leaves each, picked singly from the top, middle, and bottom of selected rows,
but not within 50 ft of the row ends. A composite (entire sampling area) canopy sample
of 75 leaves was also collected for residues. One control sample was taken at each of the
sampling periods. The whole leaves were collected in individually marked plastic bags, and
extracted immediately in the field.

A separate composite sample was collected and each of the 75 leaves was individually
blueprinted in the field. The print outlines were later removed with scissors and weighed
to determine the leaf area of residue samples.

Hands of each volunteer were rinsed with solvent at the lunch break and again at the
termination of the experiment. For ethyl parathion, 125 ml of benzene was used, while
acetone was used for monocrotophos and methyl parathion. The rinses were collected by
funnel in 200-ml screw cap sample bottles which were held in the ice chest for return to
the laboratory. T-shirts and blue jeans were placed in individual plastic bags and held in ice
chests for return to the laboratory.

2Data from U. S. Weather Station, Eloy, AZ.

 Establishment of Reentry Intervals for Cotton Fields 293

Air samples were collected with portable Mine Safety Appliance "Monitaire Samplers"
(2.83 L/min) which were attached to the belts of two subjects with the end of the air in-
take tubes pinned to the T-shirts just below the chin. After 2.5 hr of sampling, glycol
impingers were exchanged for fresh ones.

Following the field exposure to methyl parathion, rinses were made of the upper arms,
from wrist bone to shoulder, with 125 ml of acetone. Similar rinses were made from
the torso, using two ethanol-saturated 2 in. X 2 in. gauze pads, and from the dorsal leg
surfaces, each with one ethanol pad. All rinses were placed in sample bottles and held in
an ice chest for return to the laboratory. The gauzes were placed in eight-oz French square
bottles, saturated with benzene, and held in an ice chest for return to the laboratory.

Sample extraction. For determining dislodgable residue, the leaf samples were placed
in individual two-qt wide-mouth Mason jars and extracted for 30 see in the field with
900 ml of solvent, benzene for the parathions and acetone for monocrotophos. Aliquots
of extracts were held in sample jars in the ice chest and returned to the refrigerator.

Blue jeans were extracted for the parathions by one soaking in six L of benzene for at
least 30 min. T-shirts were similarly soaked in 2.5 L of benzene. Jeans and T-shirts were
extracted with acetone for monocrotophos. Aliquots were held in sample jars in the
refrigerator.

Hand rinses were held in the refrigerator and analyzed by gas chromatography in their
original condition after necessary dilutions. All air samples were extracted according to
Ware et al. (1974a).

The gauze pads were placed in funnels, squeezed and rinsed several times with benzene
and the total volumes adjusted to 250 ml to be stored at 40~

Gas chromatographic analyses. Quantitations of monocrotophos and methyl- and

ethyl parathion and their oxons were by gas chromatography, using a flame-photometric
detector sensitive to phosphorus-containing compounds. For the parathions and their
oxons, the chromatograph was equipped with a 1.52 m X 4 mm ID Pyrex glass column
packed with 100/120 mesh Chromosorb W HP containing 5% by weight of SE-30. All
chromatographic parameters were identical for all materials except that column tempera-
ture was 200~ for monocrotophos and 2100C for the remaining materials. Nitrogen gas
flow as 90 ml/min with the inlet temperature 250~

Quantitation was by peak height, compared to a standard curve made from standard
solutions containing 1.0 ng/#l of hexane for parathions, 0.2 ng//al of hexane for their
oxons, and 2.0 ng//al of acetone for monocrotophos.

Analytical recoveries from spiked samples exceeded 96% for all three pesticides.

Serum concentrations of insecticide. Sera collected before, during, and after field ex-
posures to parathions were analyzed for methyl- or ethyl parathion (but not mono-
crotophos) as described by Ware et al. (1974a).
 294 G . W . Ware et al.

Cholinesterase activities. Measurements were made by the method of Michel (1949)

and expressed as ApH/hr for both plasma and RBC. The sequential blood samples on each
subject were held in a cooler (not frozen) until all four had been collected and returned
to the laboratory. They were then tested together, using identical batches of substrate
and buffer, thus minimizing analytical variation.

Urine analysis for paranitrophenol. A pre-exposure 24-hr urine sample was collected
from each subject up to the time of entering the field. A second 24-hr sample was collected
up to approximately the same time of the morning following field entry. A third specimen
was collected in the same manner, including 24 to 48 hr after field entry. All specimens
were analyzed for PNP by the method of Elliott (1960). PNP excretion was calculated as
the product of volume times analytical PNP concentration, and has been reported as the
total PNP excreted up to 48 hr after initial field entry.

Results
Leaf residues. The dislodgable leaf residues of monocrotophos, methyl- and ethyl
parathion, and methyl and ethyl paraoxon, immediately after application and at the time
of each study are presented in Table II, expressed as mg of insecticide per m 2 o f upper
leaf surface. Residues of all three insecticides were lower when measured 1 to 3 days
after application than they were immediately post-application. Decay rates have been
shown in previous studies to be fastest for methyl parathion, slowest for mono-
crotophos (Ware e t al. 1974a, Ware e t aL 1974b).

Biomedical surveillance. None of the subjects involved in these five separate reentry
studies experienced symptoms or showed clinical signs suggesting adverse effects of ex-
posure to cholinesterase-inhibiting pesticides (miosis, bradycardia, vomiting, diarrhea,
muscle twitching, etc.). The high field temperatures and physical activity induced profuse
perspiration and dehydration, which was compensated by high rates of fluid ingestion.
The serum levels and cholinesterase-inhibiting effects of the parathions, as well as
urinary PNP excretions, corresponded qualitatively with the foliar residues, and with
measured quantities of parathion absorbed by hands and clothing. The methyl parathion
absorbed yielded readily measurable levels in the blood (Table III), and it detectably de-
pressed the red blood cell enzyme but not the plasma cholinesterase (Figures 1 and 2).

Interestingly, the 24-hr ethyl parathion residue had no effect on either cell or plasma
cholinesterase, while there were consistent declines in RBC ChE in subjects who entered
the 48-hr field (Figures 3 and 4). A possible explanation is that the field received a very
light rain before daylight of the 48-hr reentry, leaving the leaves wet, equivalent to a
moderate dew. In this particular (and only) reentry test, the subjects' jeans were soaked
almost immediately on entering the field, remained wet for approximately one hr, and
were dry within the second hr. This additional transfer of residue carried to the skin with
leaf moisture probably entailed additional absorption not experienced in previous re-
entries. Ethyl parathion was detected in the blood (Table III) at somewhat higher average
levels in the 48-hr exposure than in the 24-hr, which lends some support to the skin-
wetting hypothesis. It is relevant in this connection that a mass poisoning of field workers
 Establishment of Reentry Intervals for Cotton Fields 295

Table II. Dislodgable leaf surface residues (mg/m2 ) of methyl parathion,

ethyl parathion and monocrotophos on mature cotton plants
treated at 1.0 lb AI/acre. Coolidge, Arizona, 1973

Residue Methyl Ethyl Ethyl

age (hr) parathion parathion paraoxon Monoerotophos

0 Composite 23.7 23.3 - 12.8

Ta 22.0 31.1 0.17 24.2
M 21.0 17.5 0.21 44.7
B 3.5 14.8 0.04 37.6
x:l

12 Composite 1 6 . q .o
T 15.0 I ~'E
M 12.0]
B

U~

24 Composite 10.6
2.il 0.17
T m 3.6 0.19
M 1.6 "~ 0.13
B i. ~ 0.08

48 Composite
T
M
B
B
i.l
2.0
0.7
0.
~
m
o.12
o.12
0.08
0.06
61
6.0
4.0
1.
~o
'~

72 Composite
T
4!
7.6 ~
M
4.5
B 1.

aTop, middle and bottom canopy samples = T, M, B, respectively.

in Texas was attributable, in part, to wetness of the cotton foliage treated about 12 hr
previously with an organophosphate pesticide (Hatcher 1969).

Urinary excretions of PNP (Table III) corresponded qualitatively with blood parathion
measurements and with blood cholinesterase effects, although one might have expected
wider differences based on the disparity between methyl and ethyl blood parathion levels.
PNP excretions of some participants may have been enhanced by contact with treated
foliage prior to the day of the reentry test (see footnote to Table III).

There was a slight but consistent depression in plasma ChE in the blood of subjects
contacting the 48- and 72-hr monocrotophos residues, but only in the RBC ChE of the
296 G . W . Ware et al.

48-hr reentry. The more persistent m o n o c r o t o p h o s appears to be more readily transferred

to other surfaces than either parathion, as reflected in the hand, shirt, and pants accumu-
lations.

When pesticide was removed from skin surfaces to measure contamination, potentially
effective stores o f absorbable chemicals were reduced b y amounts up to 3.9 mg at the
noon break, and up to 1.5 mg at the termination o f the field test (methyl parathion,
Table V). Washings in the monocrotophos and ethyl parathion studies yielded smaller

Table IH. Serum parathion concentrations and 48-hr PNP

excretions during and after 5-hr fieM exposure to
cotton treated previously with methyl or ethyl
parathion (1.0 lb AI/acre)

Serum parathion (ppb) 48-hr PNP

Hr after beginning 5-hr exposure Excretion
Subject 3 7 24 (mg/48 hr)

Methyl Parathion 12-hr post-treatment

W.K. 146 100 0 1.70

M.L. 142 97 3 2.31 a
W.C. 71 70 3 1.55
E.D. 196 95 2 1.13
G.L. 227 145 4 2.12 a

Ethyl Parathion 24-hr post-treatment

G.O. 3 3 3 0.13
R.C. 5 3 1 0.09
D.M. 13 12 8 0.25
W.K. 8 6 4 0.15
E.D. 6 8 1 0.09

Ethyl Parathion 48-hr post-treatment

H.R. 7 8 4 0.15 a
M.L. 9 6 0 0.16 a
L.C. 16 2 4 0.10
G.L. 2 7 2 0.09a, b
D.L. 7 95 2 0.12

aThe urinary PNP excretions of these subjects may have been en-
hanced by prior exposure to treated foliage contacted in the course o f
their duties as cotton supervisors.
b24-hr PNP excretion for day of exposure only.
 .90 Plasma ChE 120- Plasma ChE

.80 110-

.70

.60 - 90-
1>

o
I I I I ~ 8o I I I
7o
c
r/,-.-///J 123 123~ Time of field exposure 0 r/////.4 eTA 221~ Time of field exposure
o
~.90 ~t~- RBC ChE

a. 110- ,e

.70 ~ 100-
o
, Os - ~ o
.60 90-

50 I I I I 80- I
7am loam 2pm Barn 7am lOam 2pm 8 am
| Clock hour, day of reentry | Clock hour, day of reentry
- Previous day's samples Previous day's sample

Fig. 1. Plasma and red blood cell cholinesterase responses to re- Fig. 2. Averaged blood cholinesterase response to reentry of a
entry of a cotton field treated 12 hr earlier with methyl cotton field treated 12 hr earlier with methyl parathion.
parathion. ~D
...j
 to
oo
.90 - Plasma ChE
120 - Plasma ChE

.80 -
110-
~s ..... "G-

.70 - .... - 2 Clays

100-
1 day
.60 -
.~-
> 90-

,..50- ~,
~r 80' ( "1 I
I I " I l
C~
8 ~7)77J 12~ I:Z3~ Time of field exposure
Bz~J~ E3 2:3~ Time o f field exposure
'~ 1 2 0 -
<1 e- RBC ChE
990 RBC ChE
o-110-
.80 1 day ca

100-

.70
90-

.60
I I I '
80- 1 T'--- I I
7am lOam 2pm 8 am 7am lOam 2pro 8 am
| Clock hour, day o f reentry ~ Crock hour, day of reentry
L Previous day's samples Previous day's sample

Fig. 3. Plasma and red blood cell cholinesterase responses to Fig. 4. Averaged blood cholinesterase response to reentry of a
reentry of a cotton field treated 48 hr earlier with ethyl cotton field treated 24 and 48 hr earlier with ethyl parathion.
parathion.
 Establishment of Reentry Intervals for Cotton Fields 299

amounts of surface toxicants (Tables IV and V'I). It is reasonable to suppose that regularly
employed cotton scouts engage in some skin decontamination during the work day for
purposes of personal comfort, if not for safety. In any case, dermal residues temporarily
substracted at the noon break were restored, in substantial part, as field exposure to ap-
plied pesticides was resumed in the afternoon.

The estimated respiratory exposure to vaporized insecticide is presented in Table VII.

Detectable levels were captured in all studies, with the highest level found in the field
having 12-hr methyl parathion residues. Air concentrations of pesticides (Table VII)
corresponded in relative magnitude to the foliage residue levels from which the air-borne
pesticide originated (Table II). Considering the daily oral dosage rates of the parathions
(7,5 mg for ethyl and 11.0 mg for methyl) required to depress blood cholinesterase
activities (Rider et al. 1969), it seems unlikely that absorption of these microgram
quantities over five-hr periods by any route would exert physiologic effects.

It is important to emphasize that none of the subjects monitored in this study ex-
hibited symptoms or clinical signs of organophosphate poisoning, even though: 1) pesti-
cide was present in the blood, 2) cholinesterase activities indicated effects on blood-
borne enzymes, and 3) urinary excretion of a parathion metabolite documented signifi-
cant absorption. Refined judgments as to safety of the work situation, therefore, depend
on interpretation of analytic and biochemical measures of pesticide absorption.

Given the extreme sensitivity of modern methods for detection of pesticides and
pesticide metabolites in blood and urine, a requirement that workers absorb no detectable
pesticide at all in the course of their task is probably incompatible with continued use of
organophosphates (or any other chemicalS) in agriculture. If it can be assumed that some

Table IV. Monocrotophos residues extracted

from clothing and hands following 5-hr
field exposure. Averages from 5
subjects exposed 48 and 72 hr post-application

mg after
Sample source 48 hr 72 hr

Hands
11:30 A.M. 1.16 1.71
2:30 P.M. 0.92 1.32 a

Shirts 13.50 13.40

Pants 20.20 28.70

aFour subjects
300 G . W . Ware et al.

measurable doses of these agents can be tolerated by man, the question then resolves into
a setting of quantitative criteria to provide for reasonable safety of regularly exposed
workers.

Two criteria have been suggested by occupational experience in pesticide handlers and
field workers:

1) Decrements in blood cholinesterase activities exceeding certain magnitudes have

been used as guidelines for removing workers from occupational contact with
eholinesterase-irthibiting chemicals. One authority (Gage 1967), has recommended
a 30% decrement as the limit of safety. A technical bulletin published by the
California State Health Department specifies a 50% depression in activity as one
requiting removal of a worker from pesticide contact (Bureau 1971).
2) In cases of parathion exposure, urinary PNP concentrations should not exceed two
ppm (Arterberry et as 1961, Davies et aL 1966). Assuming excretion of two L of
urine in 48 hr, this would be equivalent to a total excretion of four mg of PNP
during and following the exposure period (Table III).

Judged by the first criterion, none of the exposure situations described in these tests
presented significant hazard, as there were no individual cholinesterase depressions in

Table V. Methyl parathion residues extracted from clothing and rinses of

body surfaces following 5-hr fieM exposure. Averages from 5
subjects exposed 12 hr post-application

Sample source Methyl parathion (mg) Methyl paraoxon (mg)

Hands
11:30 A.M. 1.70 .04
2:30 P.M. .73 .08

Arms
11:30 A.M. 2.20 .06 a
2:30 P.M. .40 .06

Torso .40 .01 b

Legs .03 -001 b

Shirts 10.60 .24b

Pants 39.90 a .92 b

aFour subjects
bResidues estimated from proportion of paraoxon in hand and arm residues.
 Establishment of Reentry Intervals for Cotton Fields 301

Table. VI. Ethyl parathion residues extracted from clothing and hands following
5-hr fieM exposure. Averages from 5 subjects exposed at 24- and 48-hr
post-application

Sample source Ethyl parathion (mg) Ethyl paraoxon (mg)

24 Hr

Hands
11:30 A.M. 0.13 .004
2:30 P.M. 0.08 .004

Shirts 0.25 .014

Pants 5.71 .200

48 Hr

Hartds
11:30 A.M. 0.11 .004
2.30 P.M. 0.06 .003

Shirts 0.16 .015

Pants 2.70 .097

Table VII. Estimated respiratory dosages of volunteers working 5 hr in

cotton fields treated previously with monocrotophos, methyl- or
ethyl parathion

Estimated
Mean air respiratory
Residue age concentration of dosage
Insecticide (hr) insecticide, ng/L /.tg/5 hra

Methyl parathion 12 12.60 75.0

Ethyl parathion 24 0.62 3.7

48 0.27 1.6

Monocrotophos 48 < 0.10 < 0.6

72 <0.10 <0.6

aCalculated assuming average pulmonary ventilation of 20 L/min, and 100% ab-

sorption efficiency.
302 G . W . Ware et al.

excess of 25% of the pre-exposure value, and no averaged cholinesterase depressions

greater than 14% of pre-exposure levels.

Judged by the PNP excretion criterion, only the reentry test done 12 lar after methyl
parathion treatment generated urinary PNP approaching toxicologically significant
amounts. Actual concentrations of PNP in one-day collections ranged from 1.0 ppm to
2.3 ppm, averaging 1.5 ppm. It is important to note that even in this exposure, the de-
pressed RBC cholinesterase returned to normal within 18 hr of the end of the full ex-
posure. The ability of man to metabolize ingested methyl parathion at rates of 11 to
30 mg/day regularly without significant blood cholinesterase depression has been reported
(Rider etal. 1969 and Rider e t a l 1971).

The most complex criterion by which to assess the safety of a given exposure situation
is that of "no significant effect" on blood cholinesterase activities. The word signifi.
cant, in this case, means 1) a systematic difference or trend, improbably derived
from random sampling error, and 2) a toxicologically important effect. Small,
apparent down-trends in one or both cholinesterase activity measurements occurred
kn all of the reentry tests described herein, except the test following ethyl
parathion application by 24 hr. If statistical tests applied to data from larger subject
groups than ours confirmed the occurrence of small but real depressions of enzyme
activity at some time during or after reentry, we would have to decide whether this
demonstration should be taken as evidence of hazard. There is definite merit to this
interpretation, based on 1) the necessity for avoiding cumulative effects of the repeated
pesticide exposure to which cotton scouts are necessarily subjected, and 2) the desirability
of a certain margin of safety against such untoward occurrences as dampness of the
foliage with which scouts are in contact. The difficulty with this criterion is that it may
prove to be just as sensitive (and arbitrary) a measure of hazard as a criterion based on
detection of finite pesticide or pesticide metabolite in blood or urine.

It is clear from examination of the time-course graphs in Figures 1 to 8 that definition

of a "no effect" criterion based on cholinesterase measurements will have to include the
specific details of how an effect has been sought but not found. For example, the fre-
quency and timing of blood samples during and after field exposure is critical. Sampling
18 hr after termination of field exposure sometimes shows effects not apparent from
immediate post-exposure sampling (monocrotophos effect on plasma cholinesterase in
48- and 72-hr reentries). With some pesticides, on the other hand, (methyl parathion
effect on RBC cholinesterase in 12-hr reentry), a clear effect demonstrable just after
exposure has disappeared by the following morning.

The complexity of the cholinesterase response inspires interest in alternative methods

of assessing magnitudes of pesticide absorption by persons exposed to these chemicals
occupationally. The testing of blood for absorbed pesticide has both practical and
theoretical limitations, the latter hinging mainly on the very complex time-course char-
acteristics of materials that are rapidly metabolized and excreted.
 .90 Plasma ChE .90-] Plasma ChE
/
.80~

.70 - .70

.~ .6O

, o .50-~ --~

r
~ 40 ~ " "
r .90- RBC ChE
3:: ~77-~7~e2~ IZ3~Time of field exposure <1
O.

<1 .80-| RBC ChE .80 -

.70- ---A
C~
.70 t O

.60 .60-

.50 e~
I I t I
.50 17am lOam 2pm 8am 7'am 1barn 2pm 8Jam
/ Clock hour, day of reentry Clock hour, day of reentry
L Previous day's samples Previous day's samples
I
Fig. 5. Plasma and red blood cell cholinesterase responses to Fig. 6. Plasma and red blood cell cholinesterase responses to re-
reentry of a cotton field treated 24 hr earlier with ethyl entry of a cotton field treated 48 hr earlier with monocrotophos.
parathion.
O
 t.~
O

Plasma ChE
.90-
120 - Plasma ChE

.80-
110-

.70-
100- .... 3 days
"--o

.60-
90-
.>
,50- r

80 I I 1 "~
I I I t C
v/////,~ 173 177J~TIME OF FIELD EXPOSURE
r/////.,~ F_~ 2 2 2 - - T i m e of field exposure 8
T "6 120- RBC ChE
r RBC ChE a
,90 - 110-

.80 - 100- - o - - 3 days

.70- o 90- ~ _ 2 days -----e

v

80
7lam 10 am 2=pm 8 am 7'am 1()am 2'pm 8'am
Clock hour, day of reentry Clock hour, day of reentry
Previous day's samples Previous day's sample

Fig. 7. Plasma and red b l o o d ceil cholinesterase responses to re- Fig. 8. Averaged b l o o d cholinesterase response to reentry o f a
entry o f a c o t t o n field treated 72 hr earlier with m o n o c r o t o p h o s . c o t t o n field treated 48 and 72 hr earlier with m o n o c r o t o p h o s .
 Establishment of Reentry Intervals for Cotton Fields 305

Many new methods for quantitating urinary excretion of pesticide metabolites promise
to reduce estimation of exposure to more objective and readily interpretable parameters
(Shafik et aL 1973a, 1973b). Given some additional knowledge of the metabolic dis-
position of specific chemicals by man, it may soon be possible to use cumulated urinary
excretions to calculate actual dosages of many toxicants absorbed in the cotirse of field
exposures. The toxicologic significance of the measurements will be judged in terms of
carefully controlled studies of the effects of various toxicants over ranges of dosage
(Rider 1969, Rider 1971). Monitoring of field workers for excessive absorption of toxic
agricultural chemicals can then be patterned after methods of worker surveillance long
practiced by industrial hygienists (Linch 1974).

Conclusions
Human subjects were examined before, during and after reentry into Arizona cotton
fields at various intervals after the fields had been treated with either of three commonly
used organophosphate insecticides applied at one pound active ingredient per acre.
Neither symptoms nor clinical signs of organophosphate poisoning occurred in the course
of these field reentries, although measures of urinary PNP excretion and depressions of
blood cholinesterase activities attested to absorption of measurable and biochemically
effective amounts of toxicants in some trials.

Exposure to methyl parathion residues on foliage treated 12 hr previously (leaf residue

16 mg/m2) resulted in greater contamination of clothing and body surfaces, higher serum
parathion concentrations, higher urinary PNP excretions, and more pronounced RBC
cholinesterase depression than occurred in response to contamination with ethyl parathion
residues aged 24 and 48 hr (leaf residues 2.7 and 1.5 rag/m2, respectively).

Forty-eight hr after application at identical dosage, monocrotophos residues averaged

four times those of ethyl parathion. This larger source of toxicant generated higher levels
of contamination of skin and clothing. A clear cut effect on blood cholinesterase activities
was seen after the 48-hr post-application exposure to monocrotophos; little or no effect
appeared following contact with the 72-hr residues.

Respiratory exposure was probably not a significant route of insecticide absorption in

these field reentry tests because of the very small amounts of toxicants present in the
ambient air.

None of the plasma or RBC cholinesterase depressions observed under the conditions
of these investigations exceeded 25% in individual subjects or 14% in averaged data for
multiple subjects having the same exposure. Depressions produced by methyl parathion
showed recovery within 24 hr, while those generated by monocrotophos were more
prominent 24 hr after field exposure than during work in the field.

References
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Measurement by blood cholinesterase level and urinary p-nitrophenol excretion.
Arch. Environ. Hlth. 3, 476 (1961).
306 G.W. Ware et al.

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nosis and Treatment of Phosphate Ester. Pesticide Poisoning: Technical Bulletin
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Manuscript received April 3, 1974; accepted August 12, 1974