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Deon van der Merwe BVSc, PhD KSVDL Toxicology Section dmerwe@vet.ksu.

edu 785 532 4333

 All conceivable habitats  Incredible genetic diversity
 Over 1- 1.5 million species  Only about 70 000 species have been described  Many more

genotypes/ecotypes/chemotypes

http://herbarium.usu.edu/fungi/FunFacts/Kingfact.htm

 Mostly multicellular  Mycelium (mold) consists of branching hiphae (filaments)  Yeasts are exceptions  Usually chitin cell walls  Aerobic  Asexual/sexual reproduction  Spore-forming  Sexual reproductive structures/fruiting bodies are called mushrooms/puffballs/truffles etc.

Spore formation is key to reproductive success • Imparts genetic variation through sexual reproduction • Increases adverse condition survivability • Small and light . flavus viewed under a microscope Photos: Stella Marie M. Doyungan .easily spreads by wind/air currents Aspergillus flavus (green) growing on a corn grain A.

 Mostly saprophytic (eat dead stuff)  Good at breaking down plant material  Often symbiotic with plants/insects  Mycorrhizae  Endophytes  Can be commensal/infectious/parasitic (eg Candida spp.)  But mostly COMPETITIVE  Specialize in chemical warfare  Antibiotics eg penicillin  Mycotoxins  Human/animal poisoning is collateral damage! .

edu/.uk/smelly.gif 6CO2 + 6H2O + energy (Carbohydrate) • Smell www.jpg .squirrelldesigns.Signs of mold • Heat C6H12O6 + 6O2 www.exploratorium..co../Turkey_cooking_c.

.jpg www./images/aspercorn..ent./diplodia_ear_rot..edu/.iastate.jpg ..iastate.edu/.• Discoloration www.ipm.

Doyungan .• Caking or clumping Photo: Stella Marie M. Doyungan • Slow movement out of bins Photo: Stella Marie M.

 Economic loss  Production loss  Spoilage  Waste  Feed refusal  Health risks       Acute/chronic organ toxicities Neurotoxicity Cancer Reproductive failure Allergens/irritants Immunosuppression .

/notes/Corn/aflatoxin2.JPG  Growth stage/proliferation www..com/.ces..net. weathersavvy.jpg humidity  Generally warm and humid  Available water  Accessible nutrients  Crop damage! lh6.ca/copland/AylesIceShel.com/desert2_OPT.edu/.geomatics. Genetic potential  Environmental conditions  Aerobic conditions  Temperature and relative www..au/reslib/200710/r190166_714313./rdXiSi1bZNA/DSC03623..uottawa.ggpht...ncsu.jpg www.abc.jpg .

.gif to remain contaminated .exploratorium. Resistant to decomposition by  cooking  freezing  digestion  May end up in milk (eg Aflatoxin M)  Breakdown is often slow  Contaminated crops/food/feed tend www..edu/./Turkey_cooking_c.

and others . www. Aflatoxins  Aspergillus spp.com/MOLD12. and many others  Zearalenone  Fusarium spp.moldinspectionsinny.  Ochratoxins  Penicillium/Aspergillus spp.  Fumonisins  Fusarium spp.JPG  Trichothecenes  Fusarium/Stachybotrys spp.

spices. flavus. A.AB1  Seed crops. nominus and others  Rapid contamination  High levels within 2-7 days of inoculation  Persistent – survives processing  Many potential sources of human/animal exposure . nuts  A. oilseeds. paraciticus. A.

 Types  AB1.jpg .edu/.com/images/cancer_px/Liver_. AB2  AG1... www. AG2  AM1.brown. AM2  Metabolites of B1 and B2  Associated with milk  Potent carcinogens./HepatitisC/cirrhosis...robertsreview. Hepatotoxic  Metabolized to reactive epoxides in the liver  Causes DNA alkylation  Trout are particularly sensitive (0.4 ppb)  Tightly regulated due to health risks www.

hubpages.com/u/46516_f260.  Ochratoxin A most common  Carcinogenic. but also beans (including coffee). Nephrotoxic  Chronic renal failure typical  Evidence for human carcinogenicity weak z. Teratogenic. and Penicillium spp.  Aspergillus spp. Mostly cereals.jpg . wine etc. dried fruit.

.org/.edu/.FB1  Mostly in corn (maize)  Fusarium spp../200px-FumonisinB1.iastate. FB3  Hepatotoxic and nephrotoxic at relatively high doses in all species  Increases apoptosis www./fusarium_ear_rot.. wikimedia.png  Fumonisin B1 most common  Also FB2.jpg  Leucoencephalomalacia (LEM) in horses and pulmonary edema in pigs  Affects lipid metabolism and cardiac function ..ipm.

gif weather (optimal temp 59˚F/15˚C)  Late-harvested/overwintered grain  Fusarium spp./Vomitoxin. and others  Mostly deoxynivalenol (DON or vomitoxin)  Also diacetoxyscirpenol (DAS) and T2  Radiomimetic (bone marrow suppression)www.. Associated with grains in cool..usyd.au/.bio../9765_9959/scab-image2.gov/.ars..edu. wet bugs.usda.jpg  Triggers vomiting through effects on chemoreceptors in CNS  Decreased feed consumption and feed refusal in animals Photo: Andrew Schneider .

 Estrogenic  “False heat”.jpg ../agranco_vulvarcomp03. also other grains  Fusarium spp.engormix. infertility.com/.. Mostly corn (maize). abortion  Pigs most susceptible www.

 Lots are subject to FDA seizure  Guidance or Advisory Levels  Provide an adequate margin of safety  Not subject to enforcement by FDA . Packers and Stockyards Administration – part of the USDA) has to report violative levels to the FDA if Action Levels are exceeded. Action Levels  Must be reported to FDA  Agencies such as GIPSA (Grain Inspection.

 100 ppb .  200 ppb . swine or poultry. or mature poultry.For corn and other grains intended for finishing (i. breeding swine.For corn and other grains intended for finishing swine of 100 pounds or greater  300 ppb .  20 ppb – All other animal feeds . feedlot) beef cattle and for cottonseed meal intended for beef cattle.e. or when its destination is not known.For corn and other grains intended for immature animals (including immature poultry) and for dairy animals..For corn and other grains intended for breeding beef cattle. United States (regulated by the FDA)  20 parts per billion (ppb) .

 AM1 in milk: 0.50 ppb  Expected when AB1 in dairy cattle feed exceeds 40 ppb  Other food: 20 ppb total  Action levels in Europe are lower (but controversial)  5 ppb for AB1  4-10 ppb total depending on type of food  0.05 ppb AM1 in milk .

breeding poultry and breeding mink* 30 ppm (no more than 50% of diet)** Ruminants > 3 months old being raised for slaughter and mink being raised 60 ppm (no more than 50% of diet)** for pelt production Poultry being raised for slaughter 100 ppm (no more than 50% of diet)** All other species or classes of livestock and pet animals 10 ppm (no more than 50% of diet)** ----------------------------------------------------------------------------------------------------------------*Includes lactating dairy cattle and hens laying eggs for human consumption **Dry weight basis .Product Animal Feeds Total Fumonisins (FB1+FB2+FB3) Corn and corn by-products intended for: Equids and rabbits 5 ppm (no more than 20% of diet)** Swine and catfish 20 ppm (no more than 50% of diet)** Breeding ruminants.

Grain and grain by-products not to exceed 50% of diet Chickens Grain and grain by-products not to exceed 50% of diet Swine Grain and grain by-products not to exceed 20% of diet All other animals Grain and grain by-products not to exceed 40% of diet --------------------------------------------------------------------------------------------------------------- .------------------------------------------------------------------------------------------------------------------ Class of animal Humans Portion of diet Finished wheat products Max. DON level 1 ppm 10 ppm 10 ppm 5 ppm 5 ppm ----------------------------------------------------------------------------------------------------------------- Beef and feedlot cattle older than 4 mos.

No action. . guidance or advisory levels for ochratoxin A or zearalenone have been established by the FDA in animal feeds • These mycotoxins are handled on a case-bycase basis.

jpg .org/images/cornfield.altenergyinvestor. In the field  Prevent insect damage/plant stress  Crop rotation  Use clean seed  Plant fungus-resistant varieties  Harvest when grain moisture is low  Avoid harvesting at high rainfall periods  Deep plowing to bury spores www.

com/images/bins. In storage  Moisture control  Dry grain if wet  Monitor moisture and maintain dry conditions www.jpg  Limit mechanical and insect damage  Clean containers and equipment  Mold inhibitors can be considered .grainbinsupply.

groundnut products  Has not been formally approved by FDA . Physical methods  Removal  Separation. density segregation  Thermal breakdown  Wet/dry milling  Other types of thermal processing  Ammoniation  Ammonia gas at high temperature and pressure for 30 mins  Commercial facilities available for specific products  Cottonseed. corn.

 Hydrated Sodium Calcium Aluminosilicate (HSCAS) clay  Feed additive (Novasil®)  Reduces caking. adsorbs aflatoxin in GIT  Adsorbs aflatoxin. but NOT effective for other mycotoxins) . improves feed flow properties  Feed supplement to increase egg shell hardness  Mixed with feed.

 Aim for reliability and efficiency  Sampling points  Weighing towers. rice: 5 kg .  Distribution in bulk material is highly variable  Requires a systematic collection strategy  50-100 increments. depending on bulk mixing status  Sample size should increase with particle size  Eg groundnuts: 20 kg. conveyor belts. corn: 10 kg. storage containers etc. barges. trucks.

 Microscopy  Fluorescence under UV (not reliable!)  Mold count (serial dilution and plating)  Fair indicator of feed refusal 1: 102 1: 103 1:104 Photo: Stella Marie M. Murphy’s Rules of Thumb  “Toxic” fungi does not indicate presence of toxin  Mold-free appearance does not indicate absence of toxin  Is mold present?  Heat. smell etc. Doyungan . appearance.

 Requires well trained technicians  Equipment  Immunoassay (ELISA)  Commercial kits available  Relatively easy  Cheaper? . GC etc. HPLC. Toxin quantitation  No safety assumptions should be made without toxin quantitation  Chemical analyses  Chromatography  TLC (qualitative).