Mycotoxins and Poultry Health Natacha S. Hogan

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Mycotoxins and Poultry Health Natacha S. Hogan Department of Animal and Poultry Science, University of Saskatchewan, Saskatoon, SK Toxicology Centre, University of Saskatchewan, Saskatoon, SK

Outline Fusarium Mycotoxins and Poultry Health Post-harvest Strategies for Mycotoxins Sorting Binding/bio-protection/detoxification Current Research Broiler feeding trials In ovo studies

Fusarium Fusarium head blight disease caused by infection with Fusarium fungal species bleached heads and shriveled chalky/pinkish kernels (FDK = Fusarium damaged kernels) F. graminearum most common in western Canada in wheat Decreases grade, yield, safety, security and economical value https://www.grainscanada.gc.ca/wheat-ble/factorfacteur/fusdmg-eng.htm

Occurance of mycotoxins in North America Ochratoxin 20% Fumonisin 48% Deoxynivalenol 68% Zearalenone 37% Aflatoxin 19% Proportion of positive samples in percentage (2013)

Mycotoxins Relevant to Poultry Nutrition Mycotoxin Specific Toxins General Effects Aflatoxin (AF) Aflatoxin B1 Reduced feed efficiency, egg production, immune suppression, liver lesions

Mycotoxins Relevant to Poultry Nutrition Mycotoxin Specific Toxins General Effects Aflatoxin (AF) Aflatoxin B1 Reduced feed efficiency, egg production, immune suppression, liver lesions Ochratoxins (OTA) Ochratoxin A Poor feathering, reduced growth and egg production, kidney toxicity

Mycotoxins Relevant to Poultry Nutrition Mycotoxin Specific Toxins General Effects Aflatoxin (AF) Aflatoxin B1 Reduced feed efficiency, egg production, immune suppression, liver lesions Ochratoxins (OTA) Ochratoxin A Poor feathering, reduced growth and egg production, kidney toxicity Fumonisins (FUM) Fumonisin B1 Decreased BW gain and feed intake, liver pathology (poultry relatively resistant)

Mycotoxins Relevant to Poultry Nutrition Mycotoxin Specific Toxins General Effects Aflatoxin (AF) Aflatoxin B1 Reduced feed efficiency, egg production, immune suppression, liver lesions Ochratoxins (OTA) Ochratoxin A Poor feathering, reduced growth and egg production, kidney toxicity Fumonisins (FUM) Fumonisin B1 Decreased BW gain and feed intake, liver pathology (poultry relatively resistant) Trichothecenes T2, Zearalenone, Nivalenol Oral lesions, decreased egg production and shell quality, growth depression, immune suppression Deoxynivalenol (DON) Reduced feed intake, growth suppression, immune suppression, gut barrier

Murugesan et al. 2015 Poultry Science 00:1-18 In 2013 = 59% positive for DON (approx 4000 tests)

Murugesan et al. 2015 Poultry Science 00:1-18 In 2013 = 60% positive for DON (approx 1300 tests)

Legislated Max Tolerance Levels for Poultry Mycotoxin Commodity Level Country DON Diets for Poultry 5 ppm (mg/kg) CAN Grains destined for Poultry Diets 10 ppm USA HT-2 Diets for Poultry 0.1 ppm CAN AF Animal Feeding Stuffs 20 ppb CAN/US RECOMMENDED TOLERANCE LEVELS T-2 Diets for Poultry >1 ppm ZEA Diets for Poultry None OTA Diets for Poultry >2 ppm FUM Total ration for breeding stock Total ration for slaughter 15 ppm 50 ppm Ergot Chicks 6-9 ppm http://www.inspection.gc.ca/animals/feeds/regulatory-guidance/rg-8/eng/

Mycotoxins and Poultry Health

Poultry Performance Modern broiler chickens are selected for superior growth performance and require high quality diets to reach genetic potential (Schmidt et al., 2009; Zuidhof et al., 2014; Collins et al., 2014)

Devegowda, G., T. N. K. Murthy, and Duarte E. Diaz. "Mycotoxins: their effects in poultry and some practical solutions." The mycotoxin blue book (2005): 25-56. Under practical conditions, no poultry feed is completely free of mycotoxins The adverse effects of mycotoxins on poultry are many fold indicating a clear and persistent danger Additionally, no feed can be expected to contain only one mycotoxin

Impact of mycotoxins Direct health challenges Reduced immune response (vaccinations) Toxicity Skeletal health Reduced feed intake Reduced nutrient absorption/excretion of lipids MALABSORPTION SYNDROME Residues in meat / Eggs Condemnations / Downgrading Bruising Loss of salable product (i.e.: liver, gizzard erosion) Reproduction (loss in fertility & hatchability)

Lesions of the beak Growth and performance Lesions in the mouth Gizzard lesions Pale liver Photos: http://www.knowmycotoxins.com/index.htm

Meta-analytical analysis (Andretta et al., 2011) 98 papers (1980-2009): >1400 diets >37,000 birds Three criteria: Intoxication with mycotoxins Commercial broilers Measured performance and/or organ weights Variables: Exposure period, mycotoxin type and conc, age, BW, sex Data 1401 rows x 189 columns

Meta-analytical analysis (Andretta et al., 2011) Younger birds more affected If mycotoxins were present: Feed intake 12% Body weight 14% Ochratoxins and aflatoxins most severe Organ weights Liver 15% Kidneys 11% Lungs 9% Gizzard 3% Mortality DON - 8.8 x greater Aflatoxins - 2.8 x greater

Post-harvest Strategies for Mycotoxins

Mitigation Strategies Pre-harvest control Crop management (fungicides, fungistats) Plant genetics Biological control Post-harvest control Sorting technology Chemoprotection Chemoprevention

Sorting Damaged Kernels 1. Physical separation by hand 2. Mechanical separation 3. Near Infrared Reflectance (NIR) Spectrometry Measures reflected wavelengths (1200 2400nm) predict CP

Mycotoxin level (ppm) 30 Fraction 1 Fraction 6 25 DON 20 15 10 5 0 Unsorted F1 F2 F3 F4 F5 F6 IQ BoMill Fractions

Daily Feed Intake (g/day) Average Daily Weight Gain (ADG, g/day) Applicability in research Produce naturally contaminated diets with a range of mycotoxin concentrations from a single wheat source 300 150 200 100 Control Ad lib 100 Low Ad lib Medium Ad lib High Ad lib 50 Control Ad lib Low Ad lib Medium Ad lib High Ad lib 0 21-28 d 28-35 d 0 21-28 d 28-35 d

Current Research

1. Identify factors that influence susceptibility of poultry to adverse effects of mycotoxins 2. Develop sensitive methods for evaluating (biomarkers) exposure and toxicity in poultry 3. Test novel strategies for reducing exposure, bioavailability and toxicity of mycotoxins in production animals

Identify the critical period(s) of sensitivity to the effects of feed-borne Fusarium mycotoxins on growth performance and gut morphology in broiler chickens Starter Diet Grower Diet Treatment 1-14 d 15-21 d 22-34 d Control Control Control Control DON 1-14 d DON Control Control DON 15-21 d Control DON Control DON 22-34 d Control Control DON DON 1-34 d DON DON DON

Age 7 d 14 d 21 d 28 d 34 d Broiler Body Weight (g) Control 134.7±3.3 359.5±10.0 724.6±14.2a 1220.9±19.4a 1695.0±22.5a DON 1-14 d 129.7±3.6 330.9±14.1 673.4±17.8ab 1191.9±25.7ab 1720.0±26.4a DON 15-21 d 143.4±5.4 368.9±9.8 707.0±17.1ab 1251.7±22.9a 1717.0±28.5a DON 22-34 d 136.9±2.5 356.7±8.6 706.6±10.4ab 1117.9±17.6bc 1517.5±21.0b DON 1-34 d 131.0±2.4 332.4±7.3 650.8±13.9b 1074.6±23.3c 1433.7±24.9b Daily Feed Intake (g/day) Control 17.1±0.5 43.3±1.2 74.5±1.8 113.0±1.4 176.2±2.1ab DON 1-14 d 16.7±0.5 42.1±1.6 66.5±1.8 111.2±2.2 182.7±2.0a DON 15-21 d 17.9±0.5 44.5±0.9 73.5±2.3 115.2±2.2 176.9±1.9a DON 22-34 d 17.7±0.3 43.8±0.7 74.2±1.4 110.9±2.4 172.4±3.3b DON 1-34 d 17.4±0.8 41.6±1.1 67.3±2.2 106.7±1.9 166.5±2.7b Average Daily Gain (ADG, g/day) Control 12.8±0.5 32.1±1.1 52.2±1.2a 73.6±1.8a 96.5±1.8a DON 1-14 d 12.0±0.5 28.7±1.6 48.9±1.4ab 75.6±2.4ab 99.5±1.8a DON 15-21 d 14.0±0.8 32.2±1.4 48.3±2.0ab 77.2±2.0a 94.6±2.3a DON 22-34 d 13.0±0.4 31.4±1.3 50.0±1.3ab 58.8±2.1bc 78.5±1.9b DON 1-34 d 12.4±0.3 28.8±0.9 45.5±1.1b 60.5±1.9c 74.5±1.7b

b a

Broilers are more sensitive to moderate levels (~ 6 ppm) dietary DON during 21 35 d as indicated by reduction of feed intake, ADG and body weight Growth suppression effects of dietary DON is caused by reduction of feed intake and reduced feed efficiency The reduction of feed efficiency may be caused by alteration of small intestine histology structures

Broiler Feed Preference of Different Levels of Fusarium Mycotoxins Contaminated Diets Control (0.085 ppm DON) vs. Low (2.27 ppm DON) vs. High (5.84 ppm DON) Feeders were divided into two equal size sections Experiment diet was provided in each section Feed intake of each diet was measured on 28d Section 1 Section 2 Control Low DON Control High DON Low DON High DON

Effects of feed-borne Fusarium mycotoxins (primarily DON) on broiler feed preference during 22-28 d Control Low mycotoxins High mycotoxins Low vs. High Control vs. High Control vs. Low 0 50 100 150 Average daily feed intake (g/d) Broilers preferred the control diet over low and high mycotoxins diet. There was no preference indicated for the low vs high mycotoxin diets

Feed to gain ratio (F:G) Effects of feed-borne Fusarium mycotoxins (primarily DON) on broiler feed efficiency during 22-28 2.2 2.0 1.8 1.6 1.4 1.2 1.0 0.8 0.6 0.4 0.2 0.0 Control Low High Treatment

Acknowledgements Anhao Wang Michael Kautzman Emilie Viczko Dr. Tom Scott U of S Poultry Research Centre Prairie Diagnostic Services, Saskatoon, SK Flaman Grain Handling & Cleaning, Saskatoon, SK