Safety evaluation of a proprietary food-grade, dried fermentate preparation of Saccharomyces cerevisiae.

A safety evaluation was performed for EpiCor, a product produced by a proprietary fermentation process using Saccharomyces cerevisiae. Studies included the following assays: bacterial reverse mutation, mouse lymphoma cell mutagenicity, mitogenicity assay in human peripheral lymphocytes, and a cytochrome P450 ([CYP] CYP1A2 and CYP3A4) induction assessment as well as 14-day acute, 90-day subchronic, and 1-year chronic oral toxicity studies in rats. No evidence of genotoxicity or mitogenicity was seen in any of the in vitro or in vivo studies. The CYP assessment showed no interactions or inductions. No toxic clinical symptoms or histopathological lesions were observed in the acute, subchronic, or chronic oral toxicity studies in the rat. Results of the studies performed indicate that EpiCor does not possess genotoxic activity and has a low order of toxicity that is well tolerated when administered orally. The no observable adverse effect level (NOAEL) was 1500 mg/kg body weight (bw)/d for the 90-day study and 800 mg/kg bw/d for the 1 year study, for the highest doses tested.

Effect of different concentrations of Bacillus subtilis on growth performance, carcase quality, gut microflora and immune response of broiler chickens.

1. An experiment was conducted to study the effects of different Bacillus subtilis concentrations on productivity, carcase quality, immune response and the gut microflora of broiler chickens. 2. There were 5 treatment groups: control, with no added Bacillus subtilis supplementation; and 4 treatment groups receiving feed supplemented with different concentrations of B. subtilis. 3. Weight gain was significantly higher and the feed conversion rate was significantly better in all groups receiving feed supplemented with B. subtilis, regardless of its concentration. The groups given B. subtilis supplementation produced relatively bigger breasts, and smaller carcases and thighs, compared with the control group. 4. The appearance of increased diffuse lymphohistiocytic infiltration and solitary lymphoid follicles in the mucosa, and a stronger response to NDV vaccination, indicate increased immunological responses in chickens fed with the B. subtilis supplemented diet. 5. The higher inclusion rate of B. subtilis did not increase Lactobacillus concentrations in the ileum or in the caecum, but decreased the E. coli population significantly.

Single- and repeated-dose oral toxicity studies of citicoline free-base (choline cytidine 5'-pyrophosphate) in Sprague-Dawley rats.

The dietary supplement Citicoline free-base (choline cytidine 5'-pyrophosphate) was toxicologically evaluated in Sprague-Dawley rats using oral gavage. In an acute 14-day study, 2000 mg/kg was well tolerated. In a 90-day study, 100, 350, and 1000 mg/kg/day doses resulted in no mortality. In males, slight significant increases in serum creatinine (350 and 1000 mg/kg/day), and decreases in urine volume (all treated groups) were observed. In females, slight significant increases in total white blood cell and absolute lymphocyte counts (1000 mg/kg/day), and blood urea nitrogen (BUN) (100 and 350, but not 1000 mg/kg/day) were noted. A dose-related increase in renal tubular mineralization, without degenerative or inflammatory reaction, was found in females (all treated groups) and two males (1000 mg/kg/day). Renal mineralization in rats (especially females) is influenced by calcium:phosphorus ratios in the diet. A high level of citicoline consumption resulted in increased phosphorus intake in the rats, and likely explains this result.

Effects of oral L-carnitine, L-lysine administration and exercise on body composition and histological and biochemical parameters in pigeons.

The purpose of this study was to examine whether L-carnitine and its precursor L-lysine could have any beneficial effect in racing pigeons, and if so, whether this effect is influenced by the extent of exercise (short-distance flight: 135 km vs. long-distance flight: 580 km). Birds were divided into seven groups of animals. Group 1: negative control, no flight, no treatment, Group 2: positive control, placebo treatment before the short-distance flight, Group 3: 200 mg/day L-carnitine treatment before the short-distance flight, Group 4: 400 mg/day L-lysine treatment before the short-distance flight, Group 5: positive control, placebo treatment before the long-distance flight, Group 6: 200 mg/day L-carnitine treatment before the long-distance flight, Group 7: 400 mg/day L-lysine treatment before the long-distance flight. L-carnitine, L-lysine and distilled water (placebo) were orally administered (tube feeding) for 7 days before flight. Just after returning home, blood samples were collected and analyzed for glucose, fructosamine, cholesterol, triglycerides and thiobarbituric acid reactive substances. Pigeons were euthanized using carbon dioxide as an inhalation agent, and the whole body was subjected to proximate analysis. The status at arrival was referred to as a basis for comparison. Sex did not affect the measured parameters. As a result of the L-carnitine and L-lysine administrations, the body fat mobilization was higher during the 580 km flight, whereas no changes were noted during the 135 km flight. The main changes in the measured blood parameters were caused by the extent of exercise. This experiment considered the extent of exercise as a factor potentially modulating L-carnitine supplementation effects. In conclusion, flight distance affected several parameters but the supplements of L-carnitine and L-lysine were not effective in the tested situations.

The efficacy of valnemulin (Econor) in the control of disease caused by experimental infection of calves with Mycoplasma bovis.

Mycoplasma bovis infection was experimentally induced in groups of six young calves. A further group was uninfected and served as a control. Ten days after infection, medication with either enrofloxacin (Baytril, Bayer) or valnemulin (Econor, Novartis) was instituted via the milk replacer for a further 10 days, after which all calves were killed. Infection resulted in depression, pyrexia, inappetance and prominent respiratory signs. Arthritis occurred in two animals and two (unmedicated) animals died. At post-mortem examination extensive lesions were present in the lungs and M. bovis was re-isolated from infected unmedicated calves' lungs. Medication with either enrofloxacin or valnemulin resulted in a rapid diminution of clinical signs, restoration of appetite and reversal of weight loss. Isolation of Pasteurella multocida from the calves' lungs was suppressed by both medicaments. Valnemulin resulted in a more rapid reduction of clinical scores and eliminated M. bovis from the lungs more effectively than enrofloxacin.

Acute and subchronic toxicity studies of cryogenically-frozen, cryomilled, Pelodiscus sinensis (Japanese soft-shelled turtle--suppon) powder administered to the rat.

Sprague-Dawley rats received "cryogenically-frozen suppon" (CFS), a cryomilled product derived for the Japanese soft-shelled turtle (Pelodiscus sinensis), widely consumed for its nutritious value and medicinal properties, especially for the maintenance of normal blood pressure and insulin levels, and in women for the treatment of menopausal symptoms. In this acute study, a single limit dose of 2.0 g/kg was given po. common to 10 male and 10 female rats. No adverse effects or mortality were observed during a 14-day period and at gross pathological examination. In the subchronic study, CFS was administered as oral daily doses of 100, 350, and 1000 mg/kg administered for 97 days, resulting in no mortality, no changes in body weight, food and water consumption, hematological and serum chemistry parameters, organ weights, or gross pathology or histopathology. The only treatment related finding was a characteristic excited behavior observed in several male rats from the second or third week of treatment, distributed evenly in all male treatment groups but not affecting all males. The number of excited animals did not change over time, the syndrome occurred in all test item treated male groups with similar incidence. None of the females was so affected.

Testing the efficacy of fermented wheat germ extract against Mycoplasma gallisepticum infection of chickens.

The effect of fermented wheat germ extract (FWGE, Immunovet-HBM) was studied in chickens challenged with Mycoplasma gallisepticum. Ninety M. gallisepticum- and M. synoviae-free 3-wk-old chickens were exposed to aerosol infection of M. gallisepticum. One group (30 birds) was treated with FWGE, a second group with tiamulin, and a third group was untreated. The fourth group was exposed to PBS aerosol as a negative control. On d 9, all chickens were slaughtered and examined for the presence of gross and histological lesions, the presence of the challenge strain in the organs and specific antibodies in the serum. Body weight gains and feed conversion rates were recorded. In the groups treated with FWGE and with tiamulin, the chickens remained clinically healthy: their BW gains were 441.7 g and 446.8 g, respectively. Feed conversion ratios were 1.72 and 1.71 for FWGE- and tiamulin-treated birds, respectively. Control birds had BW gain of 480.8 g, and feed conversion ratio of 1.78. The numbers of birds with gross lesions (15 and 11, respectively) and lesion scores (25 and 25, respectively) of the FWGE- and tiamulin-treated groups were significantly lower than in the infected untreated group (25 birds, lesion score of 190). No mycoplasma was reisolated from brain, liver, spleen, heart, or kidneys of the FWGE-treated birds, and the number of mycoplasma isolations from the respiratory tract samples was less frequent (10) than from the infected untreated group (64). In addition, 35 samples from other internal organs were also positive. Twenty percent of the birds treated with FWGE showed serological response with a 5.0% reaction score, whereas in the infected untreated group, 83.3% of birds were reactors, with a 62.5% reaction score.

Effects of nickel supply on the fattening performance and several biochemical parameters of broiler chickens and rabbits.

Broiler chicken and rabbit experiments were carried out to study the effects of nickel (Ni) supplementation on growth performance and Ni metabolism. ROSS cockerels and New Zealand White female rabbits were fed a diet containing Ni in concentrations of 0, 50 and 500 mg/kg in dry matter (DM). Dietary supplementation of 50 mg Ni/kg slightly improved the body weight gain (BWG) and had a beneficial effect on the feed conversion efficiency (FCE) in broiler chickens. However, Ni added at a level of 500 mg/kg significantly (P < 0.05) reduced the BWG by 10% and resulted in significantly (P < 0.05) worse (2.3 +/- 0.2 kg/kg) FCE. The relative weight of the liver in cockerels was significantly (P < 0.05) decreased by Ni as compared to the control group (1.7 and 2.1% vs. 2.6%). The activity of AST and CHE enzymes was increased insignificantly by dietary supplementation of 500 mg Ni/kg, indicating damage of the liver parenchyma. The results of serum biochemistry were confirmed by a mild or moderate form of pathological focal fatty infiltration of the liver in broilers. Supplemental Ni of 50 mg/kg concentration resulted in non-significantly increased BWG in rabbits. Ni added to the diet at a level of 500 mg/kg reduced the digestibility of crude protein by 3-4% and that of crude fibre by 20-25% in rabbits. Approx. 98% of the ingested Ni was lost from the body via the faeces, 0.5-1.5% via the urine and approx. 1% was incorporated into the organs of rabbits. As a result of dietary supplementation of 50 and 500 mg Ni/kg, Ni accumulated in the kidneys (4.9 +/- 0.5 and 17.1 +/- 3.1 vs. 1.9 +/- 0.3 mg/kg DM), ribs (10.3 +/- 0.4 and 10.4 +/- 0.6 vs. 9.1 +/- 0.6 mg/kg DM), heart (1.4 +/- 0.2 and 2.5 +/- 0.4 vs. 1.0 +/- 0.1 mg/kg DM) and liver (1.3 +/- 0.1 and 2.2 +/- 0.2 vs. 0.9 +/- 0.05 mg/kg DM), as compared to the control animals. It can be stated that supplementation of the diet with 50 mg Ni/kg had slight but non-significant beneficial effects on the growth performance of broiler chickens and rabbits.

Effect of inorganic and organic manganese supplementation on the performance and tissue manganese content of broiler chicks.

The effects of dietary levels of manganese (Mn) in inorganic (MnO) and organic (Mn fumarate) forms were evaluated on cockerel chicks. A basal corn-soybean diet with 23 mg/kg Mn was supplemented with levels of 0, 30, 60 and 240 ppm Mn from both Mn sources. Each treatment was replicated in five pens of 10 chicks. The chicks were fed diets ad libitum from 14 to 49 days of age, after which five birds per treatment were sacrificed for pathomorphological examinations and analysis. The treatments did not exert significant effects on the body weight (BW), the feed/gain (F/G) ratio or the mortality rate. According to the necropsy findings, no growth retardation or emaciation occurred in either of the groups and the differences in the average absolute and relative organ weights were not significant (P > 0.05). Tissue analysis indicated that the tibia showed the greatest response to Mn, followed by the liver and kidney. Accumulation in the tibia was higher (P < 0.05) with supplements of 30, 60 and 240 mg/kg from both Mn sources (3.71, 3.78, 4.44, and 3.68, 4.00, 4.36 mg/kg DM, MnO and Mn fumarate, respectively) compared to the control group (3.21 mg/kg). Accumulation in the liver increased significantly (P < 0.05) only with supplements of 60 and 240 ppm independently of the Mn source (12.7, 14.2, and 14.0, 14.9 mg/kg, respectively) compared to the control (9.8 mg/kg). Similarly, kidney tissue Mn was higher (P < 0.05) only with supplements of 60 and 240 ppm (12.8, 12.8, and 13.1, 12.5 mg/kg, respectively) compared to the control (10.2 mg/kg). At the same level of supplementation of the two Mn sources there were no significant differences (P > 0.05) between the Mn concentrations of organs and tissues. Droppings sensitively reflected the intake, whereas blood plasma and feathers showed only the extreme Mn loading.

Study of soil-plant (potato and beetroot)-animal cycle of nutritive and hazardous minerals in a rabbit model.

Potato and beetroot were grown on soils previously treated with heavy metal salts. Each particular microelement had a high concentration in both potato and beetroot [cadmium (Cd) 3.7 and 55.4, lead (Pb) 8.1 and 3.0, and mercury (Hg) 5.8 and 6.8 mg/kg dry matter, respectively]. In a metabolic balance trial 16 New Zealand White rabbits were fed 50 grams of basal diet and potato or beetroot ad libitum. The apparent digestibility of major nutrients and the accumulation of the microelements in different organs were investigated. Both potato and beetroot samples of high Pb and Hg content had the significantly (p < 0.05) lowest digestibility of organic matter and nitrogen-free extract. The Cd ingested from both potato and beetroot accumulated in the kidneys and liver (2.85 and 1.48 as well as 0.459 and 0.265 mg/kg, respectively). All the microelements (Cd, Pb and Hg) accumulated in the testicles (0.196, 0.32 and 0.199 mg/kg, respectively), reducing the rate of spermatogenesis. The tissue retention of heavy metals depends not only on the element itself, but also upon the 'carrier' feedstuff.

Infectious stunting syndrome associated with disturbances of mineral metabolism and bone development in broiler chickens.

On a broiler farm with a rearing capacity of about 200,000 chickens, a disease characterised by growth retardation, variability in chick size, 'leg weakness', diarrhoea and increased mortality at 3 weeks of age occurred repeatedly, in several successive broiler flocks. Gross and histopathological findings were dominated by widening of the hypertrophic and ossification layers of the physes of long bones as well as by thickening, unevenness and defective calcification of the cartilage trabeculae. In the parathyroid gland, vacuolar degeneration of the cytoplasm of glandular epithelial cells, connective tissue proliferation and, here and there, cyst formation were seen. Additional findings included severe cerebellar oedema and neuronal degeneration. The pancreatic, myocardial and intestinal changes typical of infectious stunting syndrome (ISS) occurred only in a mild form. Four-week-old chickens exhibiting 'leg weakness' had significantly lower blood inorganic phosphate concentration and tibial ash content as compared to healthy chickens. The disease was successfully transmitted by oral administration of small intestinal homogenate from affected chickens. In a second experiment, however, the disease could not be transmitted with intestinal homogenate sterilized by irradiation. Large doses of vitamin D3 reduced the rate of growth retardation and defective calcification of bones. The digestive enzyme activities of the pancreas and small intestinal mucosa of 'infected' chickens were decreased as is typical of ISS.

Massive goitre (struma parenchymatosa) in geese.

In a goose flock consisting of 2300 birds of 6 months of age severe goitre was diagnosed. To the best of our knowledge this is the first report of naturally occurring goitre in geese, which is not related to the feeding of rapeseed meal. The major pathological findings included retarded growth and plumage development, significantly (300%) increased relative thyroid weight, fat accumulation in the mesenteric and abdominal region, and lipid infiltration of liver and kidney cells. Subsequent hormone analysis showed undetectable thyroxine (T4) levels and a dramatic drop in triiodothyronine (T3) plasma levels of the diseased geese. Thyroidal histology displayed the typical signs of struma parenchymatosa. In order to get more information about the possible causes of the goitre, 10 geese from the affected farm were transferred into the laboratories of the Central Veterinary Institute. The geese were allotted into two groups. Group I received iodine supplementation for 55 days, while the other group served as sick control (Group S). Iodine treatment caused a dramatic improvement in the birds' clinical condition except in plumage growth in Group I, while the clinical and main pathological signs of goitre remained unchanged or worsened in the untreated Group S. Contrary to this, the serum levels of thyroid hormones and responsiveness to thyrotropin releasing hormone (TRH) improved not only in Group I but also in Group S. Almost euthyroid biochemical parameters were found after 55 days of iodine treatment in Group I and, surprisingly, a considerable improvement (especially in serum T3 levels) occurred also in Group S. These findings confirm the diagnosis of goitre but also call attention to the fact that iodine deficiency was not the only factor eliciting the disorder. The underlying possible goitrogenic substance could not be traced down.

Intestinal absorption of colostral lymphocytes in newborn lambs and their role in the development of immune status.

Two model experiments were conducted to study the intestinal absorption of colostral lymphoid cells and the role of these cells in the development of immune status in newborn lambs. In experiment I, 17 lambs of 14 Merino ewes were used. Suspensions of lymphoid cells separated from the colostrum (cell density: 5 x 10(6)/ml) and blood (3 x 10(6)/ml) were labelled with technetium (Na99mTcO4) of 37 MBq/ml radioactive concentration. In three groups of lambs, 10-ml volumes of the cell suspensions were injected directly into the duodenum after laparotomy, while in a fourth group (group Ia) the same volume was administered to the animals through an oesophageal tube. The labelled cells revealed that colostral cells of the lamb's own dam are absorbed from the gastrointestinal tract and get into the newborn lamb's lymph circulation irrespective of the route of application. In experiment II, involving 40 lambs of 40 ewes, we studied the effect of absorbed colostral lymphocytes on the development of the newborn lamb's immune status. Twenty ewes (group A) each were treated with 3 ml tetanus anatoxin twice, while the remaining animals (group B) were left uninoculated. Lambs of group A (designated A2) were separated from their dams immediately after birth, then were administered, through an oesophageal tube, 10 ml of a suspension of lymphoid cells (cell density: 5 x 10(6)/ml) separated from the maternal colostrum. Subsequently, the lambs were interchanged with lambs of nonimmunized ewes of group B (designated lambs B1), i.e. were mutually put out to nursing. At three days of age, lambs of groups A1, A2, B1 and B2 were inoculated with 3 ml tetanus anatoxin, then blood samples were taken from them 5 times in a period of 27 days for comparative examination of the humoral and cellular immune reactions. The results demonstrate that lymphoid cells from the colostrum of the lambs' own dam become absorbed into the newborn lambs' lymph circulation, remain immunologically active and may transfer, besides immunological memory, also cellular activity.

Interaction of T-2 fusariotoxin and monensin in broiler chickens infected with Coccidia.

Field observations suggest that coccidiosis is a common cause of death in broiler chicken flocks fed diets containing sufficient amounts of ionophore antibiotics (monensin, narasin, etc.) and contaminated with mycotoxins, particularly with T-2 fusariotoxin. To study this phenomenon, broiler chickens fed diets containing different amounts of T-2 toxin and free from monensin, or containing a preventive dose (100 mg/kg of feed) of monensin, were infected experimentally with coccidian oocysts. In all groups fed a diet containing monensin plus T-2 toxin severe clinical symptoms of coccidiosis (blood-stained faeces etc). occurred. Deaths and retarded growth depended on the toxin dose and were considerable. The body mass gain of chicks fed a diet containing monensin and T-2 toxin but not infected with coccidia was inferior to that of groups fed diets which contained either monensin or T-2 toxin (experiment 2). On the basis of these findings a negative interaction of the two compounds is assumed. This seems to be supported by the results of experiment 3, i. e. the finding that the lethal dose of narasin, a compound closely related to monensin both in chemical structure and mechanism of action, proved to be much lower (LD50 = 102 mg/kg body mass) for chickens fed a diet supplemented with T-2 toxin than for the control chickens (LD50 = 176 mg/kg body mass). The present results suggest that the feeding of diets severely contaminated with T-2 toxin may alter the anticoccidial efficacy of monensin.

Intestinal absorption of colostral lymphoid cells in newborn piglets.

Intestinal absorption of colostral lymphoid cells was studied in 23 piglets of four sows (sows A, B, C and D). From the colostrum and blood of the sows the lymphoid cells were isolated with Ficoll-Paque and labelled with technetium (Na99mTcO4). In the 7th hour after birth, 5-ml volumes of the cell suspensions were injected, following laparotomy, directly into the stomach (piglets of sow A) or into the jejunum (piglets of sow B), whereas piglets of sows C and D received the suspensions through a naso-oesophageal tube. Cryostat sections of duodenum, jejunum and lymph node samples of piglets killed by bleeding 8 h after the treatment were examined by autoradiography. It was found that lymphoid cells present in the colostrum of a piglet's own mother were absorbed from the digestive tract and, via the lymphatic vessels, were transported to the mesenteric lymph nodes. Electron microscopy revealed that absorption took place intercellularly. Colostral cells of sows other than a piglet's own mother were observed only in the epithelial layer of the mucous membrane. The lymphoid cells isolated from the sows' blood and heat-treated colostral lymphoid cells were not absorbed. The results indicate that in the pig, an animal having an epitheliochorial placenta, the colostral lymphoid cells are absorbed from the digestive tract and, hence, they can confer an active cellular immunity on the newborn piglets.