T. K. Smith

Mycotoxins in Pet Food: A Review on Worldwide Prevalence and Preventative Strategies

Mycotoxins contaminate cereal grains worldwide, and their presence in pet food has been a potential health threat to companion animals. Aflatoxins, ochratoxin A, and Fusarium mycotoxins have been found in both raw ingredients and final products of pet food around the globe. Aflatoxin, a hepatotoxin and carcinogen, has caused several food poisoning outbreaks in dogs, and aflatoxin content is regulated in pet food in many countries. Ochratoxin A and Fusarium mycotoxins including trichothecenes, zearalenone, and fumonisins may have chronic effects on the health of companion animals. Grain processing, sampling error, analytical methods, conjugated mycotoxins, storage conditions, and synergistic interactions are common challenges faced by the pet food industry. Food-processing techniques such as sieving, washing, pearling, ozonation, and acid-based mold inhibition reduce the mycotoxin content of cereal grains. Dietary supplementation with large neutral amino acids, antioxidants, and omega-3 polysaturated fatty acids as well as inclusion of mycotoxin-sequestering agents and detoxifying microbes may ameliorate the harmful effects of mycotoxins in contaminated pet food.

Effect of dietary putrescine on whole body growth and polyamine metabolism.

Abstract Putrescine (1,4-diaminobutane) is the simplest of the mammalian polyamines. These are small, positively charged molecules which are essential for cell growth and are thought to play a role in regulation of anabolic events such as synthesis of DNA, RNA, and protein. Recent reports have indicated the potential for dietary precursor amino acids of putrescine to alter tissue putrescine concentrations. The current study was conducted to determine the physiologic significance of these effects by feeding up to flooding doses of putrescine to determine any influence on whole body growth and polyamine metabolism. A total of 96 chicks were fed purified crystalline amino acid diets containing 0.0, 0.2, 0.4, 0.6, 0.8, or 1.0% purified putrescine (four birds per pen, four pens per diet) for 14 days. The feeding of 0.2% putrescine increased growth rate beyond that of controls while further supplements reduced growth and were toxic when 0.8 and 1.0% putrescine were fed. Hepatic and muscle concentrations of ornithine increased with dietary putrescine while the effect in kidney was much less. Putrescine concentrations in liver, kidney, and muscle rose when 0.4% putrescine or more was fed. This effect was particularly obvious in muscle in which there were also increases in the concentrations of spermidine and spermine. In a subsequent similar experiment, putrescine was fed at 0.0, 0.1, 0.2, 0.3, 0.4, or 0.5% to determine the effect on the activities of the key enzymes regulating polyamine synthesis. The feeding of putrescine at even 0.1% caused a rapid reduction in hepatic ornithine decarboxylase activity while S-adenosylmethionine decarboxylase and arginase activities were not influenced by diet. It was concluded that excess tissue putrescine can be toxic to whole organisms but small, orally administered doses of this metabolite can promote growth.

Effect of Dietary Protein Deficiency and L-2-Oxothiazolidine-4-Carboxylate on the Diurnal Rhythm of Hepatic Glutathione in the Rat

Maximizing hepatic glutathione (GSH) concentration may provide greater protection against toxic compounds. A dietary supplement of L-2-oxothiazolidine-4-carboxylate (OTC), a stable derivative of cysteine, increased hepatic GSH in rats fed for 2 wk a diet deficient in protein (7.5%) but not in rats fed a diet adequate in protein (15%). Experiment 2 determined whether a dietary supplement of OTC could maintain the maximum GSH concentration over 24 h. Rats acclimatized for 5 d to a 7.5% protein diet and then fed a 7.5% protein diet supplemented with either 2.5 mmol OTC or cysteine-HCl (CYS)/100 g diet had a more rapid increase in hepatic GSH (4 and 8 h after beginning of feeding, P less than 0.05) than did rats fed an unsupplemented 7.5% protein diet. This response was not due simply to the greater supply of cysteine for GSH synthesis because supplementing the 15% protein diet with OTC or CYS had no effect on the hepatic GSH of rats acclimatized to a 15% protein diet. In experiment 3, rats acclimatized to the 7.5% protein diet had a more rapid rate of increase in hepatic GSH concentration in response to feeding than did rats acclimatized to a 15% protein diet, regardless of which diet they were fed during the 24-h period. It was concluded that in addition to cysteine availability, previous dietary protein status plays a key role in the regulation of the feeding-induced diurnal rhythm of hepatic GSH concentration in rats.

Effects of Feeding Blends of Grains Naturally Contaminated with Fusarium Mycotoxins on Small Intestinal Morphology of Turkeys

An experiment was conducted to investigate the effects of feeding grains naturally contaminated with Fusarium mycotoxins on morphometric indices of duodenum, jejunum, and ileum in turkeys. The possible preventative effect of a polymeric glucomannan mycotoxin adsorbent (GMA) was also determined. Three hundred 1-d-old male turkey poults were fed wheat, corn, and soybean meal-based starter (0 to 3 wk), grower (4 to 6 wk), developer (7 to 9 wk), and finisher (10 to 12 wk) diets formulated with control grains, contaminated grains, and contaminated grains + 0.2% GMA. Morphometric indices were measured at the end of each growth phase and included villus height (VH), crypt depth, villus width, thicknesses of submucosa and muscularis, villus-to-crypt ratio, and apparent villus surface area (AVSA). At the end of the starter phase, feedborne mycotoxins significantly decreased the VH in the duodenum, and supplementation of the contaminated diet with GMA prevented this effect. The feeding of contaminated grains also reduced (P < 0.05) VH and AVSA in jejunum, whereas none of the variables were affected in the ileum. Villus width and AVSA of duodenum, VH, and AVSA of jejunum and submucosa thickness of ileum were significantly reduced when birds were fed contaminated grains at the end of the grower phase, and supplementation with GMA prevented these effects in jejunum and ileum. No effects of diets were seen on morphometric variables at the end of the developer and finisher phases. It was concluded that consumption of grains naturally contaminated with Fusarium mycotoxins results in adverse effects on intestinal morphology during early growth phases of turkeys, and GMA can prevent many of these effects.

Effects of Feeding Blends of Grains Naturally Contaminated With Fusarium Mycotoxins on Performance, Hematology, Metabolism, and Immunocompetence of Turkeys

An experiment was conducted to investigate the effects of feeding blends of grains naturally contaminated with Fusarium mycotoxins on performance, hematology, metabolism, and immunological parameters of turkeys. The efficacy of polymeric glucomannan mycotoxin adsorbent (GMA) in preventing these adverse effects was also evaluated. Three hundred 1-d-old male turkey poults were fed wheat-, corn-, and soybean meal-based starter (0 to 3 wk), grower (4 to 6 wk), developer (7 to 9 wk), and finisher (10 to 12 wk) diets formulated with uncontaminated grains, contaminated grains, and contaminated grains + 0.2% GMA. Feeding contaminated grains significantly decreased BW gains during the grower and developer phases, and GMA supplementation prevented these effects. There was no effect of diet, however, on feed intake or feed efficiency. The feeding of contaminated grains reduced total lymphocyte counts at wk 3 (P < 0.05). Dietary supplementation with GMA increased plasma total protein concentrations compared with controls and birds fed the contaminated diet. Plasma uric acid concentrations in birds fed contaminated grains were increased at the end of the experiment compared with controls, and the feeding of GMA prevented this effect. Feeding contaminated grains significantly increased the percentage of CD4(+) lymphocyte populations during wk 6; however, there was no change in the percentage of CD8(+) and B-lymphocyte populations. Contact hypersensitivity to dinitrochlorobenzene, which is a CD8(+) T cell-mediated delayed-type hypersensitivity response, was significantly decreased after 24 and 72 h by feedborne mycotoxins compared with controls. Supplementation of the contaminated diet with GMA prevented the decrease in response after 24 h. Secondary antibody (IgG titer) response against SRBC antigens (CD4(+) T cell-dependent) was significantly decreased after feeding contaminated grains compared with controls. It was concluded that turkey performance and some blood and immunological parameters were adversely affected by feedborne Fusarium mycotoxins, and GMA prevented many of these effects.

Altered tissue amino acid metabolism in acute T-2 toxicosis.

Abstract T-2 toxin is a Fusarium trichothecene mycotoxin that has been shown to alter brain neurochemistry and eating behavior in animals eating contaminated diets. Experiments were conducted to determine the role of altered tissue amino acid metabolism in the etiology of acute T-2 toxicosis. Fasted weanling rats were orally dosed with 0 or 2.0 mg T-2 toxin/kg body weight. Blood, brain, liver, and muscle tissue were excised 4 and 8 hr after dosing, and amino acid concentrations were determined. Hepatic enlargement coupled with reduced liver concentrations of free small neutral, large neutral, and basic amino acids were seen 4 hr after dosing. Brain and muscle amino acid concentrations were largely refractory to treatment, while the plasma concentrations of tyrosine and lysine, and the sum of the basic amino acids fell. Hepatic amino acid concentrations returned to control levels 8 hr after dosing at which time aminoacidemia was seen. This was due partially to an increase in plasma concentrations of large neutral amino acids including particularly the branched-chain amino acids. A subsequent experiment was conducted to determine the effect of T-2 toxin on 14C-leucine uptake and incorporation into protein in liver slices 4 hr after dosing. Exposure to T-2 toxin reduced total (free + protein-bound) uptake of leucine due primarily to reduced incorporation of leucine into newly-synthesized hepatic protein. It was concluded that reduced amino acid uptake by liver preceded aminoacidemia in acute T-2 toxicosis, although it is not clear how this might influence subsequent changes in brain neurochemistry and behavior. [P.S.E.B.M. 1995, Vol 210]

A survey of free and conjugated deoxynivalenol in the 2008 corn crop in Ontario, Canada.

BACKGROUND Deoxynivalenol (DON, vomitoxin), one of the most important mycotoxins produced by many Fusarium species, is found as a common contaminant of crops worldwide. Recent studies have described the presence of conjugated forms of DON (glycosides and fatty acid). The aim of the current study was therefore to investigate the natural occurrence of free and conjugated DON in Canadian corn. RESULTS Free and conjugated DON was determined by gas chromatography-mass spectrometry (GC-MS) and enzyme-linked immunosorbent assay (ELISA) in 86 corn samples collected from the 2008 crop in Ontario, Canada. Free DON concentrations determined by ELISA were similar to values determined in most samples using GC-MS. Conjugated DON was detected in 72 samples. Levels of free DON ranged from 0.17 to 14.00 µg g(-1) using GC-MS. The highest levels of free DON were found in corn samples from the southern and southwestern regions of Ontario, while samples from eastern regions were less contaminated. Conjugated DON was found mainly in corn from the east-central region, with five of six samples showing high levels of conjugated DON (up to 43% increase in DON following acid hydrolysis). Low levels of conjugated DON (≤ 10% increase in DON following acid hydrolysis) were detected in the majority of corn samples from the southwestern region (nine of 19 samples) and from the central region (16 of 36 samples). CONCLUSION The current survey emphasizes the frequency of conjugated DON in Ontario grown corn and the potential challenges in understanding the hazard posed by DON-contaminated foodstuffs and feedstuffs.

Immunomodulatory Effects of Feed-Borne Fusarium Mycotoxins in Chickens Infected with Coccidia:

The potential for Fusarium mycotoxins to modulate immunity was studied in chickens raised to 10 weeks of age using an enteric coccidial infection model. Experimental diets included: control, diets containing grains naturally contaminated with Fusarium mycotoxins, and diets containing contaminated grains + 0.2% polymeric glucomannan mycotoxin adsorbent (GMA). Contaminated diets contained up to 3.8 μg/g deoxynivalenol (DON), 0.3 μg/g 15-acetyl DON and 0.2 μg/g zearalenone. An optimized mixture (inducing lesions without mortality) of Eimeria acervulina, E. maxima and E. tenella was used to challenge birds at 8 weeks of age. Immune parameters were studied prior to challenge, at the end of the challenge period (7 days post-inoculation, PI), and at the end of the recovery period (14 days PI). Total serum immunoglobulin (Ig) A and IgG concentrations in challenged birds fed the contaminated diet were higher than controls at the end of the challenge period. Serum concentration of IgA, but not IgG, was significantly decreased at the end of the recovery period in birds fed the contaminated diet. The percentage of CD4+ and CD8+ cell populations in blood mononuclear cells decreased significantly at the end of the challenge period in birds fed the control or the contaminated diet compared to their percentages prior to challenge. The pre-challenge percentage of CD8+ population was restored at the end of the recovery period only in birds fed the control diet. Interferon-γ (IFN-γ) gene expression in caecal tonsils was up-regulated in challenged birds fed the contaminated diet at the end of the challenge period. No significant effect of diet was observed on oocyst counts despite the changes in the studied immune parameters. It was concluded that Fusarium mycotoxins modulate the avian immune system. This modulation involves alteration of gene expression but apparently does not enhance susceptibility or resistance to a primary coccidial challenge.

Effects of dietary Fusarium mycotoxins on intestinal lymphocyte subset populations, cell proliferation and histological changes in avian lymphoid organs

An experiment was conducted to investigate the effects of dietary Fusarium mycotoxins on gut immunity, cell proliferation, and histology of avian lymphoid organs. The efficacy of a polymeric glucomannan mycotoxin adsorbent (GMA) was also determined. Seventy-two one-day-old male turkey poults were fed corn, wheat, and soybean meal-based diets for 21 days. Diets included control grains, contaminated grains and contaminated grains +0.2% GMA. The major contaminant was deoxynivalenol (3.9 μg/g) with lesser amounts of zearalenone (0.67-0.75 μg/g), 15-acetyl-deoxynivalenol (0.34 μg/g) and HT-2 toxin (0.078-0.085 μg/g). T- and B-lymphocyte populations and crypt cellular proliferation in duodenum, jejunum, ileum and cecal tonsil were measured immunohistochemically on day 14 and 21. Histological changes were recorded after 14 and 21 days of feeding. Feeding contaminated grains significantly increased the percentage of B-lymphocytes in ileum on day 14, and reduced (P<0.05) the percentages of CD8(+)-lymphocytes in cecal tonsil on day 21. GMA supplementation prevented these effects. The feeding of contaminated diets also caused a reduction (P<0.05) in ileal crypt proliferating cells and a significant increase in spleen secondary follicle on day 21. It was concluded that the feeding of grains naturally contaminated with Fusarium mycotoxins results in adverse effects on gut immunity and mucosal cell proliferation.

Effects of Feedborne Fusarium Mycotoxins on Brain Regional Neurochemistry of Turkeys

An experiment was conducted to investigate the effects of feeding grains naturally contaminated with Fusarium mycotoxins on brain regional neurochemistry of turkeys. The possible preventative effect of a poly-meric glucomannan mycotoxin adsorbent (GMA) was also determined. Forty-five 1-d-old male turkey poults were fed wheat-, corn-, and soybean meal-based diets up to wk 6, formulated with control grains, contaminated grains, or contaminated grains + 0.2% GMA. Deoxynivalenol was the major contaminant, and the concentrations were 2.2 and 3.3 mg/kg of feed during starter and grower phases, respectively. Concentrations of brain monoamine neurotransmitters and metabolites were measured in discrete regions of the brain including the pons, hypothalamus, and cortex by HPLC with electrochemical detection. Neurotransmitters and metabolites analyzed included norepinephrine, dopamine, 3,4-dihydroxyphenylacetic acid, serotonin (5-hydroxytryptamine, 5-HT), and 5-hydroxyindoleacetic acid (5-HIAA). The concentration of 5-HIAA and the 5-HIAA:5-HT-ratio were significantly decreased in pons after feeding contaminated grains. Dietary supplementation with GMA prevented these effects. In the pons, a significant positive correlation (r = 0.52, P < 0.05) was observed between the concentration of 5-HT and BW gain after feeding contaminated diets. The feeding of contaminated diet had no significant effects on the concentrations of neurotransmitters and metabolites in hypothalamus and cortex. It was concluded that consumption of grains naturally contaminated with Fusarium mycotoxins adversely altered the pons serotonergic system of turkeys. Supplementation with GMA partially inhibited these effects.