Effect of dietary T-2 fusariotoxin concentrations on the health and production of white Pekin duck broilers.

The effects of different dietary levels of T-2 toxin on production, biological, immunological, and pathological parameters of growing white Pekin ducks were studied to establish the "no effect" dietary concentration of, and "no effect" exposure time to, pure T-2 toxin. Day-old white Pekin ducks were randomly allotted to nine groups of 10 ducks each. One group served as a control, and no mycotoxin was added to its feed. The feeds of the experimental groups were supplemented with 0.2, 0.4, 0.6, 0.8, 1.0, 2.0, 3.0, and 4.0 mg purified T-2 toxin/kg, respectively, from Day 1 until Day 49 of the experiment. Dermatotoxic oral lesions developed in most experimental ducks within 2 d after the start of feeding T-2 toxin-contaminated feeds. The gradual disappearance of macroscopic signs indicated the development of tolerance in groups treated with the lower T-2 toxin content. No repair was found in the 3 and 4 mg/kg groups. Dietary concentrations of T-2 toxin below 0.4 mg/kg had no effect on the average weekly weight gain in the first 6 wk, but a severe decrease was found in the last week of the experiment. The 0.6 mg/kg dietary T-2 toxin had no effect on weight gain in the first 3 wk. At Week 4 and later, the weekly weight gain was significantly reduced, and the final live weight of this group was also significantly lower than that of the control. Dietary T-2 concentrations of 1 mg/kg and greater uniformly depressed growth rate. Only the 3 and 4 mg/kg groups refused feed during the first week. From Week 3 on, the feed intakes of the 0.6 to 4 mg/kg groups were usually less than that of the control group, indicating feed refusal. Serum and plasma chemical values and hematological parameters failed to show dose-dependent effects. The blastogenic response of lymphocytes to nonspecific and specific mitogens was distinctly impaired by the T-2 toxin at all levels in the feed. In the 3 and 4 mg/kg groups, the histological examination revealed lymphocyte depletion in the spleen and bursa of Fabricius.

Effect of various levels of T-2 toxin on the clinical status, performance and metabolism of growing pigs.

In two sets of experiments eight groups of seven-week-old pigs weighing about 9 kg were fed for three weeks a prestarter that contained 0.5, 1.0, 2.0, 3.0, 4.0, 5.0, 10.0 or 15.0 mg/kg of highly purified T-2 toxin. The feed of the two control groups was free from T-2 toxin. Average daily intakes of toxin by the pigs were 0.38, 0.81, 1.24, 1.43, 0.93, 0.81, 0.99 and 2.5 mg, respectively. The weight gains, the feed intakes, the extent of feed refusal, the parameters of energy and protein metabolism and the serum concentrations of calcium, inorganic phosphorus and magnesium were affected to different extents by the different doses of T-2 toxin, but the data indicated that feed consumption was reduced and the activity of aspartate aminotransferase was increased by the smallest amount of T-2 toxin tested.

Effect of various levels of T-2 toxin in the immune system of growing pigs.

Four groups of seven-week-old pigs weighing about 9 kg were fed for three weeks a prestarter that contained 0.5, 1.0, 2.0 or 3.0 mg/kg of highly purified T-2 toxin. The average daily intakes of toxin by the pigs were 0.38, 0.81, 1.24 and 1.43 mg, respectively. The experimental and control pigs were immunised with 5 ml aluminum hydroxide gel-absorbed purified horse globulin on the first and fourth days of the treatment period. Blood samples were withdrawn on days 7, 14 and 21 and used for the determination of the titre of anti-horse globulin antibody, for an in vitro lymphocyte proliferation test, using purified horse globulin, phytohaemagglutinin and concanavalin-A and for determinations of the immune complex, the cytotoxic reaction and the phagocytic activity and phagocytic index of circulating granulocytes. The samples taken on day 21 were also used to determine the erythrocyte count, the mean cell volume of the erythrocytes, the haematocrit, the blood haemoglobin concentration, the leucocyte count and the proportion of T lymphocytes. At the end of the experiment samples were taken from the thymus, spleen and mesenteric lymph nodes for histological examination. The diets that contained 2 and 3 mg T-2 toxin/kg caused a significant decrease in the red blood cell count, the mean corpuscular volume and the haemoglobin concentration. A significant decrease in the leucocyte count and the proportion of T lymphocytes was observed in all the treatment groups. There were also dose-dependent, significant decreases in antibody formation and in the blastogenic transformation of lymphocytes, and mild to moderate reactive processes were observed histologically in the lymphoid organs.

Effects of T-2 toxin treatment on the egg yield and hatchability in geese.

Ten groups of one-year-old geese, each including 10 layers and 3 ganders, were treated with T-2 toxin at doses of 0, 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.8, 1.0 and 3.0 mg/kg body weight/day for 18 days by injection into the stomach. At the toxin dose of 0.1 mg/kg/day, the egg yield remained unchanged while the hatching rate slightly decreased. T-2 toxin administered at a dose of 0.2 mg/kg/day decreased the hatching rate and the egg yield. The toxin dose of 0.3 mg/kg/day resulted in a 50% reduction of the hatching rate. The mortality rate was significant at toxin levels higher than 0.8 mg/bwkg/day; however, 30% of the birds survived even the 3.0 mg/kg/day toxin dose.

Perinatal oestrogen syndrome in swine.

Perinatal oestrogen syndrome (F-2 fusariotoxicosis occurring at the perinatal age) was studied in large pig herds and in animal experiments. The disease markedly lowered the conception rate of sows and gilts, and increased the number of repeat breeders. Litter size decreased and the number of stillbirths rose. Both the newborn piglets and the stillborn fetuses showed swelling of the vulva and teats and oedematous infiltration of the perineal region, ventral part of the abdomen and umbilicus, often accompanied by exudative-crusted inflammation, then necrosis of the teats. The number of piglets with splayleg and trembling increased. Gross and histopathological examination revealed enlargement of the ovary and uterus, with signs of follicle maturation in the ovary, glandular proliferation in the endometrium and epithelial proliferation in the vagina in addition to oedema and hyperaemia. In newborn piglets, the signs of hyperoestrogenism could be induced also experimentally, by feeding an F-2 toxin containing diet to pregnant sows. Intrauterine toxin effect was found to be primarily responsible for inducing the syndrome in newborn piglets. Because of its lower quantity, F-2 toxin excreted in the sow's milk is assumed to have a secondary role.

Pathomorphological changes caused by T-2 trichothecene fusariotoxin in geese.

T-2 trichothecene fusariotoxin was administered to 110 laying geese and 33 ganders in the active egg production period through a tube at 2-day intervals on a total of 10 occasions. After the treatment, the geese were subjected to detailed pathomorphological examination. In the ovaries of layers, a cessation of follicle maturation and follicle degeneration dependent on the toxin dose were observed, accompanied by ovulation and consequent peritonitis in the birds that died and in some of those killed by bleeding. Additional findings included involution of the oviduct, lymphocyte depletion, necrosis and amyloidosis in the spleen, catarrhal enteritis, signs of colloid stasis in the thyroid and large numbers of secretory granules in the cytoplasm of the adrenaline-producing cells of the adrenal gland. In the ganders, toxin administration did not reduce the intensity of spermatogenesis but in the spleen, intestine and endocrine glands it caused changes similar to those seen in the layers.

Characterisation and effects of a xylanase enzyme preparation extracted from Thermomyces lanuginosus cultures.

This paper describes the production of an enzyme preparation from the fungus Thermomyces lanuginosus. Thermal resistance, pH stability and lignocellulolytic activity of the enzyme preparation high in xylanase were studied on a variety of grains and forages. The enzyme preparation preserved more than 70% of its original xylanase activity for 4 and 1 h at 60 and 70 degrees C, respectively. The xylanase activity remained over 80% when the preparation was incubated for 30 min at pH 4.5. In vitro digestibility studies indicated that the enzyme digested 7.5, 8.5 and 8.0% of the dry matter (DM) of barley meal, wheat bran and oat meal samples, respectively. When applying 60-min incubation, 7.5, 7.3 and 8.4% of DM of the oat straw, alfalfa hay and triticale straw was digested, respectively. When the time of digestion was increased to 360 min, the sunflower hull showed 15.8% DM digestibility.

Effects of a non-starch polysaccharidase enzyme preparation from Thermomyces lanuginosus on energy and protein metabolism and milk yield of dairy cattle.

Non-starch polysaccharides (NSPs) form an integral part of the cell walls in plants and represent considerable available energy when degraded into absorbable mono-, di-, tri- and oligosaccharides. The ruminal microflora hydrolyses a good part of NSPs, however, recently there have been attempts to enhance the rate of utilisation by using external polysaccharidase enzymes. In the present study the effects of an enzyme preparation (Rumino-Zyme) high in xylanase activity were studied on ruminal volatile fatty acid (VFA) concentration, parameters of energy and protein metabolism, milk yield, feed conversion ratio (FCR) and body condition score of high-yielding dairy cows. A lignolytic enzyme preparation produced by the thermophilic fungus Thermomyces lanuginosus was applied in the present experiment and fed to dairy cows at 34 g/day dosage in the period between calving and the 110th day of lactation. This preparation increased VFA concentration in the rumen from about 32 days after calving and onward. Increased VFA concentration was followed by an about 5 to 10% increase in milk production and an almost 0.1% increase in butterfat production. Increased VFA concentration produced more balanced energy metabolism in the experimental cows as indicated by the lower incidence rate of hyperketonaemia, and lower acetoacetic acid and non-esterified fatty acid (NEFA) concentration in the blood of the experimental cows. Aspartate aminotransferase (AST) activity was tendentiously higher in the control group and the proportion of cows that had AST activity higher than 100 U/l was also higher in the control group. Both control and experimental cows showed balanced protein and acid-base metabolism throughout the experiment. Enhanced VFA concentration contributed to an improvement in energy balance in the experimental cows with a resultant improvement of feed intake and feed utilisation. Due to the more balanced energy metabolism postparturient body condition loss of the treated cows was reduced.

Simultaneous detection of some fusariotoxins by gas-liquid chromatography.

A method was developed for the simultaneous determination of 6 Fusarium mycotoxins (deoxynivalenol, diacetoxyscirpenol, HT-2 toxin, T-2 toxin, fusarenon-x, and zearalenone). Cereal samples were first extracted with ethyl acetate, then with a mixture of methanol-water. The crude extracts were combined and purified by silica gel chromatography. The purified extract was reacted with BSTFA (N,O-bis(trimethylsilyl)-trifluoroacetamide) to form the derivative, and chromatographed on an SE-52 wall-coated open tubular column. Kováts' retention index was determined for the 6 mycotoxins investigated. Recoveries and standard deviations were determined for pure toxin mixed in cereal. Recovery was 70-80%; relative standard deviation was 10-18%. The method developed was applied to different cereal samples.

Evaluation of mycotoxin-contaminated cereals for their use in animal feeds in Hungary.

In the period between December 5, 1991 and September 17, 1998, 760 maize, 367 wheat, 119 soybean, 222 barley, 85 bran, 32 triticale, 60 oat, 14 rye and 22 sunflower samples were investigated for the presence and concentration of seven fusariotoxins (T-2 toxin, zearalenone, deoxynivalenol, nivalenol, diacetoxyscirpenol, HT-2 toxin, fusarenone-X) and OTA. The comparison of analytical data with those of the relevant literature revealed that although the incidence rate and/or concentration of Fusarium mycotoxins and OTA in Hungarian-grown cereals is occasionally considerable, the position of the country is not worse that the average of countries. Our findings indicate that soybean tends to be good substrate for trichothecene-producing fungi and the rate of contamination is regarded as substantial. The commodities were assorted into one of three quality categories. The proportion of objectionable samples was only 3.0, 2.2, 2.3 and 1.7% in maize, wheat, barley and soybean samples, respectively. However, this low rate of objection might still be a source of great economic loss. The proportion of objectionable samples was much higher in the case of bran, oat and triticale (7.1, 6.7, and 6.3%, respectively). The results of the present investigation indicate a need for regular screening for mycotoxins of importance and individual appraisal of each commodity from the point of their use in animal feeds.

Isolation of macrocyclic and non-macrocyclic trichothecenes (stachybotrys and fusarium toxins) from the Environment of 200 III Sport Horses.

Satratoxins H and G, verrucarin J, and roridin E were isolated from the bedding straw of 200 sport horses exhibiting typical symptoms of stachybotryo-toxicosis. At the same time, the oat feed consumed by the horses contained non-macrocyclicFusarium trichothecenes: T-2 toxin and diacetoxyscirpenol.

Isolation of satratoxins from the bedding straw of a sheep flock with fatal stachybotryotoxicosis.

During a period of several weeks, more than 100 sheep died at a Hungarian farm. The animals exhibited fleece loosing, and hemorrhaging was the most important autopsy finding. Pasteurella haemolytica was cultured from various organs. The bedding straw was abundantly covered with Stachybotrys atra, and removal of the straw stopped the disease. Methanol extraction of the bedding straw followed by solvent partitioning, column chromatography, preparative thin-layer chromatography, and high-pressure liquid chromatography led to the isolation of satratoxins G and H, which were characterized by thin-layer chromatography, high-pressure liquid chromatography, and mass spectroscopy. This is the first isolation and characterization of toxins from a field sample of material responsible for an outbreak of stachybotryotoxicosis.

Preliminary communication: examination of the harmful effect to fetuses of fumonisin B(1) in pregnant sows.

Three sows were fed a diet mixed with Fusarium moniliforme fungal culture from the 107th day of pregnancy until parturition. Fumonisin B(1) toxin was administered to two sows (sows 1 and 2) in a daily dose of 300 mg for an additional 7 days subsequent to parturition, i.e., for a total of 14-16 days. The third sow (No. 3) was given the toxin in the same daily dose only until parturition, i.e., for 7 days in total. There were no symptoms observed in any of the sows. Two piglets were taken from each of the three sows and sacrificed immediately after parturition, i.e., prior to the first suckling. After 24 h, two additional piglets were taken for slaughter from each of the litters, which by then had access to colostrum. Finally, on the 7th day postparturition another two piglets per litter were sacrificed and material obtained from them was processed for examination. It was established that fumonisin B(1) present in the Fusarium moniliforme culture resulted in damage to the fetuses in utero. Of the changes indicating toxic effect, intraalveolar, subpleural, and interstitial pulmonary edema of various degrees of severity could be detected in the piglets sacrificed immediately following parturition and before the first suckling. Pathological changes were observed in the histopathological sections of the liver, and increases in the activities of plasma aspartic acid transaminase (AST), gamma glutamyl transferase (GGT) and alkaline phosphatase (AKLP), higher than physiological levels were detected. The serum-free sphinganine/sphingosine ratio, considered a bioindicator of fumonisin B(1) toxicosis, varied in accordance with the degree of severity of the changes which occurred. The values obtained were found to be between 0.29 and 0.36 in the cases of severe pulmonary edema, and between 0.20 and 0.24 for the cases of mild pulmonary edema. In the piglets of the sows fed the toxin for an additional 7 days subsequent to parturition and which were born with severe pulmonary edema, mild pulmonary edema could be detected after colostrum suckling, 24 h, and 7 days after parturition. The SA/SO value of the serum in these two piglets was 0.19 and 0.20, while at the same time AST, GGT, and ALKP values higher than physiological levels were measured. In the milk samples taken from sows 1 and 2 and examined after 24 h and after 7 days FB(1) was detected in quantities of 18.0-27.5 ppb. There were no changes observed on the seventh day in the piglets of the third sow, the diet of which contained no toxin after parturition. However, as the piglets of the third sow demonstrated only mild effects of pulmonary edema it is not possible to establish with certainty a postpartum cause-effect relationship between fumonisin in colostrum and pulmonary edema.

Hydrogen breath test in small intestinal malabsorption.

Jejunal biopsy and hydrogen breath test were performed in 57 children, 34 having coeliac disease and 23 with other forms of malabsorption. In children affected by coeliac disease there was a gradually increasing incidence of positive findings with the H2 breath test as villous damage progressed. In the group of subtotal villous atrophy age dependence was also observed, the majority of positive results occurring below three years of age. In the non-coeliac group the most frequent cause of the positive finding was Giardia lamblia infestation. Among 27 cases with lactose malabsorption confirmed by a positive hydrogen breath test only 11 had diarrhoea. The test proved to be useful in differentiating between the contaminated intestine syndrome and malabsorption due to reduced absorptive surface.