Aflatoxins

In Search of
One Health
Solutions

Esther Giezendanner*
Blair Budd*
In collaboration with the North Carolina One Health Collaborative

© 2012 North Carolina One Health Collaborative

 What are Aflatoxins?
• Toxins produced by the fungi Aspergillus flavus and Aspergillus
parasiticus
• Aflatoxins are secondary fungal metabolites.
• Aflatoxin types include B1, B2, G1, G2.
• B1 is most prevalent and toxic aflatoxin.

Microscopic view : spore

formation of Aspergillus Chemical structure of aflatoxin B1

• Detection:
• Fluorescence can be used to detect presence of Aspergillus on
crops
• Biomarkers are used to detect aflatoxin exposures in humans
 The One Health Approach
• “The concept of One Health is an
evolving, interdisciplinary way of
approaching complex health issues by
recognizing the interconnectedness of
human health, animal health and the
environment.Ӡ
• The goal of this presentation is to
illustrate that aflatoxins are a worldwide
health problem that could benefit from a
One Health approach
 Aflatoxins and One Health
• Aflatoxins cause health problems around the world
including areas as diverse as Africa, Southeast Asia,
Western Pacific, East Mediterranean and Latin America
where as many as 5 billion persons may be exposed
• Aflatoxins have a negative economic impact on
agriculture through reduced marketing options for
crops and adverse health effects on livestock9
• A One Health solution to the aflatoxin problem must:
• Be collaborative across scientific disciplines and
leverage shared knowledge
• Address cultural and societal issues related to plant
agriculture, human and animal health.
• Utilize advances in biotechnology and medicine
 Examples of Aflatoxin-Related Events
• United Kingdom
• 1960s: Turkey X disease and bird die-offs attributed to ‘syndrome X’
• Both incidents were the result of acute aflatoxicosis
• United States
• 1998: Crop contamination
• Aflatoxin contamination of maize (corn) in the south-eastern U.S. led
to rejection rates of corn of up to 50%.
• Aflatoxin contamination reached 1500 ppb (5 times the 300 ppb
highest acceptable limit in animal feed set by the U.S. Food and
Drug Administration)
• 2006-2007: Crop contamination
• Drought conditions and moisture stress led to aflatoxin on corn in
Missouri which caused rejection of some harvested corn by buyers
• Kenya
• 2004-2005: Acute Aflatoxicosis and human mortality
• Aflatoxin contamination of maize caused over 150 human deaths
Aflatoxins: Human, Animal, and Environmental
Interactions
Environment:
extreme Fungal
drought, growth /
moisture, heat, aflatoxin
compromised production
plants

Contamination
of human &
animal food

Human
consumption
Breast milk
In utero Milk Animal
eggs
consumption
Infants
 Aflatoxins and Environmental
Conditions
• Conditions favoring aflatoxin formation include:
• High temperature
• High humidity
• Presence of external plant stressors:
• Periods of drought
• Insect infestation
• Soil conditions conducive to Aspergillus growth
• High organic content
• High moisture
 Aflatoxins and Food Production

Major crops affected by aflatoxins include maize (corn) and

groundnuts (peanuts). Agricultural practices can be
modified to reduce aflatoxin production / contamination.
Farming practices Storage practices
• irrigation • drying techniques
• pesticide use • processing, such as
• time of harvest shelling peanuts
• exposure to pests

Aspergillus on maize Drying maize

 Aflatoxins and Acute Human
Health Effects
Acute aflatoxicosis can be fatal.
Presenting symptoms are determined by amount
of toxin consumed.
Clinical symptoms in humans include:
• Abdominal pain
• Vomiting
• Pulmonary edema
• Liver necrosis
 Chronic Aflatoxin Exposure
and Human Health
• Carcinogenicity
• Liver cancer is a serious consequence of long-term
exposure to aflatoxins.
• Hepatitis B infection may exacerbate the effects of
aflatoxin exposure and vaccination against Hepatitis B
can help reduce carcinogenicity of the toxin.
• Other consequences of chronic exposure include
decreased immune and reproductive function.
• Children chronically exposed may experience growth
failure.
• Infants may be exposed through breast milk.
• The fetus may be exposed during pregnancy if the
mother consumes aflatoxins.
No level of aflatoxin exposure is considered
safe for humans.
 Aflatoxins in Wildlife
Migratory birds are often exposed.
• Birds are a highly sensitive species.
• Birds consume aflatoxins when they feed on
contaminated crops.

Contaminated crops such as corn, fed to hunting

game as attractants, may result in wildlife
population decline; this is a One Health issue since
humans are responsible for this exposure.
 Aflatoxins in Farmed Animals
• Poultry
• Highly sensitive
• Aflatoxin toxicity impairs uptake of essential nutrients as well as causing
tissue damage
• Ruminants
• Ruminants are relatively insensitive; however, aflatoxin exposure can cause
growth impairment in young or lactating animals.
• Metabolites in milk and related dairy products
• Aflatoxin consumed by cows is excreted in milk as the M1 metabolite.
• The M1 metabolite can be absorbed by calves or humans causing growth
failure.
• The M1 metabolite also remains present in milk-based products such as
cheese and yogurt.
• Fish
• When farmed fish are accidentally fed contaminated grains, large die-offs may
occur.
• Rainbow trout are highly sensitive

Animal deaths and reduced productivity from aflatoxin exposure can have
significant negative ‘economic’ impact in addition to the negative health
outcomes for those who consume contaminated animal products.
 Aflatoxins in Companion Animals
• Aflatoxin has been found in pet foods in
North and South America, Europe, and
the Middle East.
• Examples of pet food recalls in the U.S.
• Diamond Pet Foods Company recalls
dry dog food recalled in 2007
• 76 dogs died
• Cause: aflatoxin-contaminated
corn
• O’Neals Feeder Supply, Inc.’s Arrow
brand dry dog food recalled in 2011
• Aflatoxin contamination found
and food recalled before health
effects were reported
 Aflatoxin Pathogenicity in
Laboratory Animals
• Experimental studies in animals are used to study the
mechanism of acute and chronic human health effects
of aflatoxins
• Species sensitivity varies
• Rodents
• Rats demonstrate malignant transformation of cysts in livers (
• Rat studies were used to determine that there is ‘no safe level’ of
aflatoxin exposure.
• Adult mice may be more resistant, but aflatoxins are highly
pathogenic in young rats
• Primates
Rhesus, Cynomolgus, and African Green monkeys given
aflatoxin for at least 2 months have an overall liver tumor incidence of
50%; for those surviving more than 6 months the incidence was 19%3
 Strategies for Reducing
Aflatoxin Exposure
• Regulations
• Agricultural production quality control
• Food processing and crop storage safety
• Early recognition and medical management of
health effects
• Educational outreach
• Community
• Individual
 Regulation
• Although no level of aflatoxin is considered “safe”, some
exposure to aflatoxins is probably unavoidable at low levels
• At least 99 countries have aflatoxin regulations
• Even with regulations in place, lack of adequate testing in some
countries may prevent enforcement
• Safety standards
• U.S. safety regulations include aflatoxin limits for human foods
(maximum 20 µg/kg) and animal feeds (300 ppb)
• E.U limits are stricter than in the US (maximum 10 µg/kg for
direct human consumption)

Aspergillus on peanuts
 Allowable Aflatoxin Levels
in Human Foods26

Amount Food type

20 ppb Foods in general

0.5 ppb (aflatoxin M1) Milk

20 ppb Peanuts and peanut products

20 ppb Pistachio nuts

20 ppb Brazil nuts

Allowable Aflatoxin Levels in Animal Feeds26

Amount Feed Type

20 ppb For corn and other grains intended for immature animals
(including immature poultry) and for dairy animals, or when its
destination is not known
20 ppb For animal feeds, other than corn or cottonseed meal;
100 ppb For corn and other grains intended for breeding beef cattle,
breeding swine, or mature poultry
200 ppb For corn and other grains intended for finishing swine of 100
pounds or greater
300 ppb For corn and other grains intended for finishing (i.e., feedlot)
beef cattle and for cottonseed meal intended for beef cattle,
swine or poultry
 Farming and Storage Practices that
Prevent Aspergillus Growth
• Pre-harvest • Harvest
• Pest management for • Prevent compromise
insects (particularly to the crop by
soil insects), weeds, harvesting when
and nematodes mature
• Planting date • For maize, harvest
• Irrigation early to prevent
• Crop rotation or completion of the
fertilization Aspergillus life cycle
• Use of drought • Post-harvest
tolerant and locally • Proper drying
adapted varieties • Storage in a dry place
 Post-Exposure Management
and Prevention of Disease
• Dietary interventions
• Increase dietary diversity
• Consume compounds that impair aflatoxin absorption
• NovaSil™: clay that binds aflatoxin in the gut
• Fermentation: Lactobacillus may bind aflatoxin
• Hepatitis B vaccination
• Hepatitis B and aflatoxin exposure interact to increase
risk of liver cancer
• Vaccination against Hepatitis B reduces liver cancer
rates by 45-50%
 Prevention: Economic Challenges
• Cost is a major challenge since many areas where aflatoxin
contaminated crops are staples lack financial resources
• Losses due to aflatoxins cost $900 million annually in Indonesia,
the Philippines and Thailand alone28
• Stakeholders who benefit are not necessarily those who bear the
costs
• Growers: carry the cost burden of many interventions
• Consumers: benefit most from interventions
• Government: mediate between growers and consumers, by
regulation or funding of interventions, to improve public health
• Local markets provide very little financial incentive to reduce
aflatoxin contamination
• A One Health approach will try to balance the needs of the
growers to make a sustainable living with protecting consumers
from aflatoxin contamination by managing environment.
 The Benefits of an Interdisciplinary One
Health Approach
• Educating stakeholders on the interconnectedness of
humans, animals and the environment is the first step in
preventing aflatoxin-related health issues

Human

One
Health

Environment Animal
 Innovative Solutions to the Aflatoxin
Problem
• Biocontrol
• Atoxigenic Aspergillus
• Atoxigenic strains of Aspergillus
compete with toxigenic strains,
preventing production of aflatoxin

• Aflasafe™ is one such strain

• Use in Nigeria resulted in an 80%
reduction in aflatoxin levels
• May not need to be reapplied
annually
• United Nations Industrial
Development Organization is
supporting the use of Aflasafe™
Toxigenic and atoxigenic
strains of Aspergillus
 Production Recommendations
• Improved Farming Practices
• Irrigation reduces stress on plants
• Pest management prevents crop damage
• Crop rotation
• Harvest at the appropriate time
• Storage
• Proper drying pre-harvest and storage in a dry place

• Using proper farming and storage practices are simple

interventions that can be gradually introduced

A One Health approach recognizes that human

impacts on the environment play an important role
in Aspergillus growth and the production of
aflatoxins.
 Human Consumption
Recommendations
• Exposure reduction
• Dietary modification
• Diet diversity reduces aflatoxin exposure and
improves overall human nutrition
• Dramatic dietary alterations may not be
affordable or culturally acceptable in many
areas
• Education on sources of aflatoxin can alert
consumers to risks
 Human Health Recommendations
• Post-exposure management
• Hepatitis B vaccination
• Decreases rates of liver cancer
• Complementary to decreasing aflatoxin exposure
• This intervention can be funded and implemented
by governments without burdening growers
• NovaSil™ clay
• Prevents intestinal uptake through adsorption of
aflatoxins
• Prevention is the primary long-term goal
 Sources
1. Hussein HS, Brasel JM. Toxicity, metabolism, and impact of mycotoxins on humans and animals. Toxicology. 2001;167(2):101-34.
2. European Union. Commission Regulation (EC) No 1881/2006 of 19 December 2006 setting maximum levels for certain contaminants in foodstuffs. 2006.
http://eur-lex.europa.eu/LexUriServ/LexUriServ.do?uri=CONSLEG:2006R1881:20100701:EN:HTML Accessed April 24, 2012.
3. Kensler TW, Roebuck BD, Wogan GN, Groopman JD. Aflatoxin: a 50-year odyssey of mechanistic and translational toxicology. Toxicol Sci. 2011;120 Suppl
1:S28-48. Epub 2010 Sep 29.
4. Shephard GS. Aflatoxin analysis at the beginning of the twenty-first century. Anal Bioanal Chem. 2009;395(5):1215-24. Epub 2009 May 31.
5. Aspergillus flavus. http://www.aspergillusflavus.org/aflavus/ Accessed April 24, 2012.
6. Wild CP, Turner PC. The toxicology of aflatoxins as a basis for public health decisions. Mutagenesis. 2002;17(6):471-81.
7. Bommakanti AS, Waliyar F. “Importance of Aflatoxins in human and livestock health."Aspergillus and Aflatoxin in Groundnut. International Crops
Research Institute for the Semi-Arid Tropics. 2000. Available at: http://www.icrisat.org/aflatoxin/health.asp Accessed April 1, 2012.
8. Hollis PL. Preventing aflatoxin begins on the farm. Southeast Farm Press. 2001. http://southeastfarmpress.com/preventing-aflatoxin-begins-farm
Accessed April 1, 2012.
9. Brandenburg, RL. William Personal communication. April 27, 2012.
10. Krenke R, Grabczak EM. Tracheobronchial manifestations of Aspergillus infections. ScientificWorldJournal. 2011;11:2310-29. Epub 2011 Nov 20.
11. Strosnider H, Azziz-Baumgartner E, Banziger M, et al. Workgroup report: public health strategies for reducing aflatoxin exposure in developing countries.
Environ Health Perspect. 2006;114(12):1898-903.
12. Liu Y, Wu F. Global burden of aflatoxin-induced hepatocellular carcinoma: a risk assessment. Environ Health Perspect. 2010;118(6):818-24. Epub 2010
Feb 19.
13. Sweets, Laura E., and J. Allen Wrather. "Aflatoxin in Corn." University of Missouri Delta Research Center. College of Agriculture, Food, and Natural
Resources. 2009. http://aes.missouri.edu/delta/croppest/aflacorn.stm Accessed Mar. 9, 2012.
14. Olufunmilayo GO, Oyefolu AB. Natural occurrence of aflatoxin residues in fresh and sun-dried meat in Nigeria. Pan Afr Med J. 2010;7:14. Epub 2010 Nov
19.
15. Prandini A, Tansini G, Sigolo S, Filippi L, Laporta M, Piva G. On the occurrence of aflatoxin M1 in milk and dairy products. Food Chem Toxicol.
2009;47(5):984-91. Epub 2007 Oct 13.
16. Pandey I, Chauhan SS. Studies on production performance and toxin residues in tissues and eggs of layer chickens fed on diets with various
concentrations of aflatoxin AFB1. Br Poult Sci. 2007;48(6):713-23.
17. Khlangwiset P, Shephard GS, Wu F. Aflatoxins and growth impairment: a review. Crit Rev Toxicol. 2011;41(9):740-55. Epub 2011 Jun 28.
18. Wagacha JM, Muthomi JW. Mycotoxin problem in Africa: current status, implications to food safety and health and possible management strategies. Int
J Food Microbiol. 2008;124(1):1-12. Epub 2008 Jan 24.
19. Cotty PJ, Jaime-Garcia R. Influences of climate on aflatoxin producing fungi and aflatoxin contamination. Int J Food Microbiol. 2007;119(1-2):109-15.
Epub 2007 Aug 14.
20. William JH, Grubb JA, Davis JW, et al. HIV and hepatocellular and esophageal carcinomas related to consumption of mycotoxin-prone foods in sub-
Saharan Africa carcinomas and mycotoxin prone foods. Am J Clin Nutr. 2010;92:154-60.
 Sources (cont.)
21. Manary MJ. Local production and provision of ready-to-use therapeutic food (RUTF) spread for the treatment of severe childhood malnutrition. Food
Nutr Bull. 2006;27(3 Suppl):S83-9.
22. Perkins, JR. Supplemental feeding. 1991. Texas Parks and Wildlife Department Fisheries & Wildlife Division.
http://www.tpwd.state.tx.us/publications/pwdpubs/media/pwd_bk_w7000_0033.pdf Accessed April 24, 2012.
23. Leung MC, Díaz-Llano G, Smith TK. Mycotoxins in pet food: a review on worldwide prevalence and preventative strategies. J Agric Food Chem.
2006;54(26):9623-35.
24. "Toxic Pet food may have killed dozens of dogs." MSNBC News. 5 Oct 2006. http://www.msnbc.msn.com/id/10771943/ns/health-pet_health/t/toxic-
pet-food-may-have-killed-dozens-dogs/ Accessed Mar. 12, 2012.
25. Sagman, Mike. "Arrow Brand Dog Food Recall." Dog Food Reviews. Dog Food Advisor, 2012. http://www.dogfoodadvisor.com/dog-food-recall/arrow-
brand-dog-food-recall/ Accessed April 24, 2012.
26. U.S. Food and Drug Administration. Guidance for Industry: Action Levels for Poisonous or Deleterious Substances in Human Food and Animal Feed. 2000.
http://www.fda.gov/Food/GuidanceComplianceRegulatoryInformation/GuidanceDocuments/ChemicalContaminantsandPesticides/ucm077969.htm
Accessed April 24, 2012.
27. European Union. Commission Regulation (EC) No 1881/2006 of 19 December 2006 setting maximum levels for certain contaminants in foodstuffs. 2006.
http://eur-lex.europa.eu/LexUriServ/LexUriServ.do?uri=CONSLEG:2006R1881:20100701:EN:HTML Accessed April 24, 2012.
28. Takumi, Yoshizawa. "Mycotoxins and Food Safety Current Situation of Food Contamination, Regulations and Risk Assessment for Mycotoxins." Foods &
Food Ingred J Jpn. 211.12 (2006): 1018-1026. Web.
2012. http://www.ffcr.or.jp/zaidan/ffcrhome.nsf/7bd44c20b0dc562649256502001b65e9/b71bcd8b8ad963b54925725000229627/$FILE/211(12)5.pdf
Accessed April 12, 2012.
29. United States. Federal Department of Agriculture. Inspections, Compliance, Enforcement, and Criminal Investigations. 2005.
http://www.fda.gov/ICECI/ComplianceManuals/CompliancePolicyGuidanceManual/ucm074598.htm Accessed April 24, 2012.
30. Lawley, Richard. "Aflatoxins." Food Safety Watch. N.p., Nov 2007. Web. http://www.foodsafetywatch.com/public/482.cfm Accessed March 9, 2012.
31. Munkvold G, Hurburgh C. "Aflatoxin in Corn." Iowa State University. 2009. http://www.extension.iastate.edu/Publications/PM1800.pdf Accessed May 7,
2012.
32. Khlangwiset P, Wu F. Costs and efficacy of public health interventions to reduce aflatoxin-induced human disease. Food Addit Contam Part A Chem Anal
Control Expo Risk Assess. 2010;27(7):998-1014.
33. Groopman JD, Kensler TW, Wild CP. (2008) Protective interventions to prevent aflatoxin-induced carcinogenesis in developing countries. Annu Rev
Public Health. 29:187-203. Review. PubMed PMID: 17914931.
34. Afriyie-Gyawu E, Wang Z, Ankrah NA, Xu L, Johnson NM, Tang L, Guan H, Huebner HJ, Jolly PE, Ellis WO, Taylor R, Brattin B, Ofori-Adjei D, Williams JH,
Wang JS, Phillips TD. NovaSil clay does not affect the concentrations of vitamins A and E and nutrient minerals in serum samples from Ghanaians at high
risk for aflatoxicosis. Food Addit Contam Part A Chem Anal Control Expo Risk Assess. 2008 Jul;25(7):872-84.
35. Marroquín-Cardona A, Deng Y, Garcia-Mazcorro J, Johnson NM, Mitchell N, Tang L, Robinson A 2nd, Taylor J, Wang JS, Phillips TD. Characterization and
Safety of Uniform Particle Size NovaSil Clay as a Potential Aflatoxin Enterosorbent. Appl Clay Sci. 2011 Dec;54(3-4):248-257.
36. Schmale III, David G, and Gary P Munkvold. "Mycotoxins in Crops: A Threat to Human and Domestic Animal Health: Economic
Impact." Mycotoxins (2012). APSnet. Web. Available at: http://www.apsnet.org/edcenter/intropp/topics/Mycotoxins/Pages/EconomicImpact.aspx 21
Apr 2012. Accessed April 24, 2012.
37. Atherton, G. "New biopesticide Aflasafe™ may solve Kenya's ongoing maize contamination problem.” The Aspergillus Website blog. N.p., 12 Jul 2010.
Web. Available at: http://aspergillusblog.blogspot.com/2010/07/new-biopesticide-aflasafe-may-solve.html Accessed Mar. 12, 2012.
 Review Slide
• Aflatoxins are mycotoxins , produced by the fungus,
Aspergillus; they are toxic to humans and animals.
• Aflatoxin toxicity ranges from acute effects (death) to
chronic effects (liver cancer).
• Exposure of children to aflatoxins can retard growth.
• Wildlife, agricultural animals, laboratory animals, and
pets are all susceptible to the negative health effects of
aflatoxin exposure
• Environmental conditions, such as moist soil and warm
temperatures, promote Aspergillus growth and
aflatoxin production.
 Review Slide (continued)
• Regulations for aflatoxin contamination in foods vary from
country to country.
• Farming and storage practices that protect plants from
stress help reduce aflatoxin production and crop
contamination.
• Increased dietary diversity and Hepatitis B vaccination can
help protect humans against the effects of aflatoxins.
• A challenge to aflatoxin regulation is the cost of rejected
crops and mitigation efforts borne by food producers and
other stakeholders
• Control of aflatoxin contamination and exposure is an
opportunity to demonstrate the value of a One Health
approach for solving a complex problem involving humans,
animals, and the environment.
 Image Credits
• Slide 1: “Broadcasting Aflasafe in maize field.” © 2008 International Institute of Tropical Agriculture Image Library.
http://www.flickr.com/photos/iita-media-library/6846617887/ Used under a Creative Commons Attribution-
NonCommercial license: http://creativecommons.org/licenses/by-nc/2.0/deed.en
• Slide 1: “Grain Sampling Program” © 2010 Texas AgriLife Research photo by Blair Fannin.
http://www.flickr.com/photos/agrilifetoday/6432128831/ Used under a Creative Commons Attribution-
NonCommercial-NoDerivs license: http://creativecommons.org/licenses/by-nc-nd/2.0/deed.en
• Slide 1: “Aflatoxin-contaminated groundnut kernels” © 2008 International Institute of Tropical Agriculture Image
Library. http://www.flickr.com/photos/iita-media-library/4684917072/ Used under a Creative Commons Attribution-
NonCommercial license: http://creativecommons.org/licenses/by-nc/2.0/deed.en
• Slide 2: “B0004539 Aspergillus spore formation (conidia), phase contrast” ©2003 Wellcome.
http://www.flickr.com/photos/wellcomeimages/5987578301/ Used under a Creative Commons Attribution-
NonCommercial-NoDerivs license: http://creativecommons.org/licenses/by-nc-nd/2.0/deed.en
• Slide 2: Public domain: http://commons.wikimedia.org/wiki/File:Aflatoxin_B1.png
• Slide 8: “Grain Sampling Program” © 2010 Texas AgriLife Research photo by Blair Fannin.
http://www.flickr.com/photos/agrilifetoday/6432128831/ Used under a Creative Commons Attribution-
NonCommercial-NoDerivs license: http://creativecommons.org/licenses/by-nc-nd/2.0/deed.en
• Slide 8: “Aspergillus infected maize” © 2011 International Institute of Tropical Agriculture Image Library.
http://www.flickr.com/photos/iita-media-library/5781888774/ Used under a Creative Commons Attribution-
NonCommercial license: http://creativecommons.org/licenses/by-nc/2.0/deed.en
• Slide 13: Based on the work: “Dog food” 2008 photo by notto86.
http://www.flickr.com/photos/nickotto/2164557362/ Used under a Creative Commons Attribution-ShareAlike license:
http://creativecommons.org/licenses/by-sa/2.0/deed.en
• Slide 16: “Aflatoxin-contaminated groundnut kernels” © 2008 International Institute of Tropical Agriculture Image
Library. http://www.flickr.com/photos/iita-media-library/4684917072/ Used under a Creative Commons Attribution-
NonCommercial license: http://creativecommons.org/licenses/by-nc/2.0/deed.en
• Slide 23: “Atoxigenic and toxigenic strains” © 2003 International Institute of Tropical Agriculture Image Library.
http://www.flickr.com/photos/iita-media-library/5761978714/ Used under a Creative Commons Attribution-
NonCommercial license: http://creativecommons.org/licenses/by-nc/2.0/deed.en