John Tuite

Ochratoxin A Toxicosis in Swine

Ochratoxicosis was induced in young female swine by a diet contaminated with a rice culture of Aspergillus ostianus that contained ochratoxin A and by daily oral doses of 2.0 or 1.0 mg/kg body weight pure ochratoxin A. Mycotoxicosis was characterized early by depression and reduction in feed intake and loss of body weight, followed by diarrhea, polyuria, polydipsia and dehydration. The pigs given pure ochratoxin A were dead or moribund in 5 to 6 days. Packed cell volume, hemoglobin, total plasma protein, and blood urea nitrogen were increased. Progressive leukocytosis, neutrophitia and moderate left shift in the differential count occurred. Concentrations of lactic dehydrogenase, isocitric dehydrogenase and glutamic-oxalacetic transaminase in the serum and urine were increased by the fourth to sixth day, but only the increase in urinary concentrations was significant. Gross findings included dehydration, enteritis, pale tan discoloration of the liver, and edema and hyperemia of the mesenteric and other lymph nodes. Microscopic lesions were most frequent and severe in the kidney and gastrointestinal tract. Necrosis of renal tubular epithelium was most frequent in the convoluted tubules. Many renal tubules were dilated. The intestinal lesions were focal and necrotizing and occurred in mostanat omic regions. Fatty change was demonstrated in most of the livers. In lymphoid tissues the changes were edema, hyperemia and focal necrosis of lymphocytes within germinal centers and around follicles.

Citrinin mycotoxicosis in beagle dogs

Abstract Three trials were completed in which young beagle dogs were either fed a citrinin-containing culture of Penicillium citrinum (Trial I) or were administered pure citrinin either ip (Trial II) or orally (Trial III). Dogs fed the fungal culture and those given 20 or 40 mg pure citrinin/kg body weight developed renal disease. Dogs given 2·5 and 5 mg/kg remained clinically normal and without renal lesions. Clinicopathological evidence of renal damage included a rise in blood urea nitrogen, glucosuria, proteinuria, lowered urinary specific gravity, increased urinary activities of lactic dehydrogenase, glutamic-oxalacetic transaminase and isocitric dehydrogenase and the presence of numerous necrotic renal tubular epithelial cells in the urinary sediment. Emesis and intestinal intussusception occured in the dogs given 20 or 40 mg/kg ip. Renal lesions consisted of degeneration and necrosis of the tubular epithelium, most prominently in the thick segment of Henle and in the distal convoluted tubules.

Investigations of the toxic effects in mice of certain species of Penicillium

Abstract Several species of Penicillium isolated from corn were tested for toxicity in mice. Corn cultures of the several species were incubated either at 24 or 16°. After the fungi were treated with chloroform, the cultures were dried and ground for mixing with either chicken mash or a purified diet at a ratio of 2:1 or 1:1 mold culture to diluent. Generally, corn cultures were more toxic when incubated at 24°. Penicillium oxalicum and P. viridicatum were clinically toxic, reducing weight gains and increasing the number of deaths. Other isolates which were less toxic, especially when fed at 1:1 ratio, included P. citrinum, P. variabile, P. purpurogenum, P. expansum, P. multicolor, P. frequentans, P. puberulum , and P. urticae . Only P. viridicatum consistently produced gross and microscopic lesions. About 40% of test mice were jaundiced and had green urine positive for bile pigments. Hepatic lesions including greenish discoloration and mottling of surface due to focal areas of necrosis were observed in most of the mice. Microscopic lesions included disseminated focal areas of necrosis, bile ductule cell hyperplasia, cholangitis and periductular fibrosis often accompanied by hypertrophy and hyperplasia of ductal epithelium. Six isolates of P. viridicatum were tested, and only the NRRL culture failed to elicit the gross and microscopic lesions as described above. A portion of the toxic activity of the P. viridicatum culture appeared to be thermolabile as weight gains were greater and mortality less in group of mice fed the corn culture subjected to steaming.

Ochratoxin A and Citrinin Induced Nephrosis in Beagle Dogs I. Clinical and Clinicopathological Features

Ochratoxin A and citrinin, both mycotoxins, were given separately and combined to young Beagle dogs for 14 days. Ochratoxin A, 0.1 and 0.2 mg/kg, was given by capsule, and citrinin, 5 and 10 mg/kg, was dissolved in ethanol and given by intraperitoneal injection. Clinical signs of toxicosis in dogs given 10 mg/kg citrinin and the higher combined doses included anorexia, retching, tenesmus, weight loss, prostration and death. Severity of the clinical disease and mortality were increased when the mycotoxins were combined, which indicated synergism. The clinicopathological abnormalities reflected renal damage, in that glutamic oxaloacetic transaminase and lactic dehydrogenase increased in the urine of the dogs with clinical signs of poisoning. Serum lactic dehydrogenase was increased in dogs given 10 mg/kg citrinin. Cellular and granular casts, ketones, protein and glucose were in the urine of dogs given large doses of citrinin alone or combined with ochratoxin A. Serum concentrations of sodium, potassium and chloride decreased in the dogs given high doses of each group.

Ochratoxicosis in Beagle Dogs. II. Pathology

The gross lesions in 23 young male Beagle dogs given daily oral doses of 0.2–3.0 mg/kg body weight ochratoxin A consisted of moderate to severe mucohemorrhagic enteritis of the cecum, colon, and rectum; tonsillitis: and enlargement of lymph nodes, which were edematous, hyperemic and focally necrotic. Histopathologic alterations in the kidneys consisted of necrosis and desquamation of epithelial cells mainly in, but not limited to, proximal convoluted tubules. Many proximal and distal convoluted tubules contained eosinophilic, granular casts. Necrosis of lymphoid tissues was a prominent feature of the mycotoxicosis and focally occurred in, but was not limited to, germinal centers in the spleen, tonsils, thymus, and lymph nodes and the lymphoid nodules of the jejunum, ileum, cecum, colon, rectum and nictitating membrane. Necrosis did not occur in bone marrow. Hepatic alterations consisted of slight to moderate centrilobular necrosis and fatty change and occurred mainly in dogs receiving 0.2–0.3 mg/kg body weight ochratoxin A daily for 11–14 days.

Species of Penicillium occurring in freshly-harvested and in stored dent corn kernels.

SUMMARY During 1964-1968 in Indiana, samples of dent corn kernels were obtained from fields at harvest, from cribs, bins and experimental storage tests. The Penicillia which emerged from surface-disinfected kernels, plated on malt-salt and on PDA containing Tergitol NPX and chlortetracycline, were isolated and identified. Penicillia were found consistently in unstored corn (6.4% of kernels infected), in crib samples (13.4%), and in commercial samples of poor quality (21%). Penicillium oxalicum and P. funiculosum were the chief species isolated from unstored kernels, although P. cyclopium was found consistently in small amounts, while P. cyclopium, P. brevi-compactum and P. viridicatum were the chief species isolated from stored kernels. The following were infrequent from either or both sources: P. luteum, P. frequentans, P. implicatum, P. charlesii, P. purpurogenum, P. multicolor, P. variable, P. citrinum, P. steckii, P. urticae, P. palitans, P. puberulum, P. chrysogenum, P. digitatum, P. janthinellum, P. expansum, P. granulatum. The economic loss caused by fungus deterioration in the storage of

Ochratoxicosis in Beagle Dogs. 1. Clinical and Clinicopathological Features

Rice culture of Aspergillus ochraceus that contained ochratoxin A or pure crystalline ochratoxin A was administered orally daily to 23 young male Beagle dogs. Six other young male Beagles served as controls. Most dogs were killed in extremis on or before day 14 after receiving a daily oral dose of ochratoxin A, 0.2–3.0 mg/kg body weight. Consistent clinical features of the mycotoxicosis were anorexia, weight loss, emesis, tenesmus, passage of clots of blood-stained mucus from the rectum, rectal temperature up to 107°F, tonsillitis, dehydration and prostration. The prominent clinicopathological features of the toxicosis were a reflection of renal damage. The urine had a specific gravity as low as 1.004 and contained granular casts, necrotic renal epithelium, and concentrations above normal of protein, glucose, lactic dehydrogenase, isocitric dehydrogenase, leucine aminopeptidase, glutamic-pyruvic transaminase, glutamic-oxalacetic transaminase, and alkaline phosphatase. Serum concentrations of these enzymes did not change. Concentrations of blood urea nitrogen remained, with two exceptions, within the normal range, but the concentrations of blood glucose declined terminally. Serum concentrations of the liver-specific enzyme, γ-glutamyl transpeptidase and sulfabromophthalein dye retention time did not indicate hepatic dysfunction.

Ochratoxin A and Citrinin Induced Nephrosis in Beagle Dogs II. Pathology

Beagle dogs were given ochratoxin A (0.1 and 0.2 mg/kg) and citrinin (5 and 10 mg/kg) alone and in two dose combinations for 14 days. The gross lesions included focal peritonitis and intestinal intussusceptions in dogs given citrinin. Changes in the kidneys of dogs given ochratoxin A were degeneration and necrosis with desquamation of tubular epithelial cells, primarily in the straight segment of the proximal tubules. Dogs given 10 mg/kg citrinin had similar changes in the distal tubules and collecting ducts. Dogs given combined doses of citrinin and ochratoxin A had degeneration and necrosis in proximal and distal tubules, and in thin segments and the collecting ducts; there were desquamated cells and granular casts in the lumina. Dogs given ochratoxin A had necrosis of lymphoid tissues in the spleen, tonsil, thymus, peripheral lymph nodes and lymph nodules of the ileum, colon and rectum. There was ulceration of the mucosa of the intestine in dogs given large combined doses of ochratoxin A and citrinin.

Citrinin mycotoxicosis in broiler chickens

Citrinin, a mycotoxin, was administered to chickens by crop gavage in dimethylsulphoxide-70% ethanol (3:1, v/v) or mixed with the diet (commercial starter mash) in four separate trials. The single-dose oral LD50 value in 7-day-old male broiler chicks was found to be 95 mg/kg. The administration of seven daily doses of citrinin equal to a half and three-quarters of the single-dose LD50 produced no additive toxic effects. The development of tissue lesions was studied in chicks killed sequentially after administration of the LD50 in a single dose. Watery diarrhoea and increased water consumption were the most consistent signs of citrinin mycotoxicosis. Histopathological alterations were most prominent in the kidneys and included degeneration and necrosis of tubular epithelial cells of both the proximal and distal convoluted tubules. Hepatic lesions were multiple foci of necrosis and haemorrhage and the severity was correlated with citrinin dose. Lymphoid necrosis and depletion were found in the bursa of Fabricius, thymus, caecal tonsil and spleen in birds given a single and multiple oral doses of citrinin. When chickens were given 100, 250 or 500 ppm citrinin in the diet for 3 wk they showed the clinical signs of citrinin mycotoxicosis and their kidneys were enlarged but no histopathological alterations were found.

Emesis and refusal of feed in swine associated with Gibberella zeae-infected corn.

Abstract In 1965, 28 northern Indiana counties reported refusal of corn by swine. Samples consistently yielded high numbers of kernels infected with Gibberella zeae (Fusarium graminearum) . “Red streak” was common in the corn, but did not contribute to refusal of feed. Water and methanol extracts of the corn contained an emetic substance, presumed to be a metabolite of G. zeae , effective on oral or parenteral administration. Extracts from 35 g of infected corn were adjusted to 100 ml and the volumes required to cause emesis in pigs calculated. Oral doses were 1.28–4.55 ml per kg, intraperitoneal doses were 1.74–2.02 ml per kg, and intravenous doses were 0.59–0.83 ml per kg of body weight. Emesis was independent of refusal in experimental pigs which suggests that unrelated substances caused the 2 effects. Uterine hypertrophy did not occur when 16 samples of infected corn were fed to immature female laboratory animals. Therefore, it is proposed that 3 distinct active metabolites are produced by G. zeae; one anabolic and causing uterine hypertrophy, one with emetic activity, and one causing refusal to consume infected feed.