Purified T-2 toxin was fed in a diet to broiler chicks from one day to 9 weeks of age in concentrations of 0, 0.2, 0.4, 2.0, and 4.0 p.p.m. to evaluate the subacute toxicity of the toxin. There were 36 chicks per T-2 concentration and 12 chicks were necropsied at 3 week intervals.The weight gain of chicks fed a diet containing 4 p.p.m. T-2 toxin was significantly (P < 0.05) less than that of chicks fed the control diet or a diet containing 0.4 p.p.m. T-2 toxin while feed conversion was similar between treatments. The weight gain was most sensitively affected by the toxin during the first 3-week period. The heart weight was significantly (P < 0.05) increased in chicks fed 1 and 2 p.p.m. T-2 toxin, but not in chicks fed 4 p.p.m. The liver and gizzard weights were not influenced by the dietary T-2 toxin.Oral lesions were observed in chicks fed the diet containing 4 p.p.m. T-2 toxin from the second week and nearly 75% of the total chicks in this treatment developed oral lesions after the 3rd week. The lesions were a circumscribed proliferative yellow caseous plaque type, and occurred at the margin of the beak, mucosa of the hard palate and angle of the mouth, and tongue. The severity and incidence were proportional to the amount of T-2 toxin in the diet. There were no outward clinical or pathological signs except that a few birds fed 4 p.p.m. T-2 toxin showed necrotic lesions on the mucosal lining of the gizzard.No abnormal changes were found in the bone marrow. As compared with control chicks, there was no marked change in blood components (hematocrit, hemoglobin, erythrocyte, leukocyte, leukocyte differential count) in T-2 treated chicks. In chicks fed high concentrations of T-2 toxin, the serum levels of alkaline phosphatase, LDH and cholesterol tended to decrease while uric acid increased.

Purified T-2 toxin was given to broiler chicks and laying hens in single oral doses by intubation into the crop to evaluate acute toxic effects. The 10-day median lethal dose for one-day-old broiler chicks in two experiments was estimated as 5.03 ± 0.25 and 5.25 ± 0.21 mg./kg. body weight. The 10-day median lethal dose for 8-week-old broiler chicks and laying hens was 4.97 ± 0.25 and 6.27 ± 0.42 mg./kg. body weight, respectively. Death of birds occurred within 48 hours after T-2 toxin administration. Within 4 hours after receiving T-2 toxin, birds developed asthenia, inappetence, diarrhea and panting. Coma was observed in birds given high lethal doses of T-2 toxin. The abdominal cavities of birds given lethal doses contained a white chalk-like material, which covered much of the viscera.Sublethal doses of T-2 toxin resulted in a decrease of weight gain and feed consumption which were proportional to the amount of the toxin given. No obvious abnormal changes in the bone marrow were noted in surviving birds. The blood components (hematocrit, hemoglobin, erythrocyte, leukocyte, SGOT, SGPT, serum LDH, serum alkaline phosphatase, serum total protein, serum cholesterol and serum uric acid) determined at 10 and 30 days after T-2 toxin administration were not significantly changed or not consistently changed in 8-week-old broiler chicks and laying hens by T-2 toxin.Other than necrotic visible lesions on the mucosal surface of the gizzard and crop in a few high dose birds, no salient pathological or clinical signs were observed at 10 and 30 days after T-2 toxin administration.

Purified T-2 toxin was fed to S.C.W.L. hens at levels of 0 (control), 0.5, 1.0, 2.0, 4.0 and 8.0 p.p.m. of an otherwise balanced diet. Feed consumption, egg production and shell thickness were significantly (P less than 0.05) decreased in hens fed 8 p.p.m. as compared with control hens. The fertility and progeny performance were not depressed by feeding T-2 toxin, but the hatchability of fertile eggs of hens fed 2 and 8 p.p.m. was significantly (P less than 0.05) lower than that of hens fed the control diet. The weights of liver, heart, gizzard and spleen were not influenced by T-2 toxin. Serum levels of alkaline phosphatase, LDH and uric acid of hens fed high concentrations of T-2 toxin were greater than those of control hens. SGPT in hens fed 8.0 p.p.m. was lower when compared with control hens. No outward changes in hematocrit, hemoglobin, erythrocyte, leukocyte and differential leukocyte counts were noted with feeding T-2 toxin. Most hens fed T-2 toxin developed oral lesions: circumscribed proliferative yellow caseous plaques at the margin of the beak, mucosa of the hard palate and angle of the mouth, and tongue. The incidence and severity of lesions were proportional to the dietary level of T-2 toxin. The only other lesion observed in necropsy examination at the end of the experiment was the small mucosal ulcer in the anterior portion of the gizzard in hens fed high levels of T-2 toxin. Microscostrointestinal tract, etc.) revealed no significant pathological change except the necrotic lesions in the gizzard and crop.

Purified zearalenone was fed to broiler chicks from 1 to 21 days of age at levels of 0 (control), 10, 25, 50, 100, 200, 400, and 800 ppm in a diet in order to evaluate its subacute toxicity. Weight gain, feed consumption, and feed/gain ratio were not affected at any of the concentrations administered. Weights of the liver, heart, spleen, testicles, oviduct, comb, and bursa of Fabricius were similar between control and zearalenone treated groups. The leukocytes of chicks fed 400 and 800 ppm zearalenone decreased as compared with that of control chicks due to the decrease in total number of lymphocytes. Hematocrit, hemoglobin, erythrocytes, and serum levels of calcium, phosphorus, total protein, alkaline phosphatase, and cholesterol were not affected by zearalenone. The incidence and size of ovarian cystic development were greater in chicks fed zearalenone, although cysts were found in some of the control group. There were no prominent signs in gross necropsy examinations other than hypertrophy of the oviduct in some birds fed 800 ppm zearalenone. The results of this study suggest that broiler chicks are tolerant of zearalenone and less sensitive than swine and turkey poults.

A sensitive method is described for the determination and confirmation of zeranol and zearalenone, as well as their isomers and metabolites, in edible animal tissue. The analytes are extracted from tissue with methanol, hydrolyzed enzymatically, cleaned up by acidbase partitioning, determined by liquid chromatography (LC) with electrochemical (EC) detection, and confirmed by gas chromatography/ mass spectrometry (GC/MS). LC analysis is performed by isocratic elution with a buffered mobile phase using a Nova-Pak reverse-phase C,„ column with amperometric EC detection at +0.90 V. Capillary GC/MS analysis of the trimethylsilyl derivatives provides mass spectral confirmations.