Effects of Bacillus cereus var. toyoi on immune parameters of pregnant sows

Changing immune parameters during pregnancy have previously been reported in humans and cattle, and have been suggested to contribute to increased susceptibility to infections. However, data regarding immune parameters during pregnancy in sows are rare. In this study, we investigated the peripartal immune status of sows using phenotypical (FACS analysis) as well as functional (proliferation assays, cytokine analysis) parameters of peripheral blood mononuclear cells (PBMCs) in pregnant sows. In previous studies, we reported a modulation of the immune system after feed supplementation of the probiotic Bacillus cereus var. toyoi in piglets [Schierack, P., Wieler, L.H., Taras, D., Herwig, V., Tachu, B., Hlinak, A., Schmidt, M.F., Scharek, L., 2007. Bacillus cereus var. toyoi enhanced systemic immune response in piglets. Vet. Immunol. Immunopathol. 118, 1–11]. Here, we extended these previous studies to include investigations of possible probiotic effects on the peripartal immune status of sows and their reproductivity. We show that immune parameters of sows change during pregnancy, the proliferative response of PBMCs to several bacterial antigens in control animals decreased from days 90 to 30 ante partum. Relative numbers (%) of CD3+CD8+, CD4+, cytotoxic T, CD14+ and CD21+ cells were reduced compared to non-pregnant sows. In contrast, the proliferative response of PBMCs of probiotic-treated sows increased during pregnancy. Bacterial antigens primarily stimulated the proliferation of naïve CD21+ cells and the relative CD21+ cell numbers were elevated in the probiotic group in the absence of effects on other immune cell populations. The clinical and microbial status of both control and probiotic sows was similar, excluding pre-existing health problems or infections as responsible for the immunological changes, and feed supplementation also had no significant effects on reproductivity. The results suggest that the probiotic B. cereus var. toyoi can alter the proliferative response of lymphocytes and affects the immune cell population ratios of pregnant sows. How and to what extent this may affect health and reproductivity should be the focus of further studies.     

Influence of the probiotic Bacillus cereus var. toyoi on the intestinal immunity of piglets

In a feeding trial, sows and piglets were fed with the probiotic bacterium Bacillus cereus var. toyoi as a feed additive, and the effects on immune cell populations were examined. The development of the gut immune system was determined for piglets at the ages of 14, 28, 35 and 56 days post partum. Tissue samples of the Jejunum and the continuous Peyer's patch were used for enumeration of intraepithelial lymphocyte populations by fluorescence activated flow cytometry and fluorescence microscopy. Both independent methods of investigation led to similar results: the population of intraepithelial CD8+ T cells was significantly enhanced in the probiotic group piglets (p ≤ 0.05), and the numbers of γδ T cells tended to be higher in the intestinal epithelium (p < 0.1) at the time of weaning (day 28). Lamina propria lymphocytes were also influenced by the treatment. Application of B. cereus var. toyoi resulted in significantly more CD25+ lymphocytes and γδ T cells in the probiotic group post-weaning. The occurrence of pathogenic Escherichia coli serogroups was also less frequent in the feces of piglets from the probiotic group. The finding that the CD8+ T cell population in the intestinal mucosa showed changes on day 28 indicated that the influence of B. cereus var. toyoi supplementation on the intestinal immune system started before weaning, an observation supported by changes in the intestinal microflora observed during the suckling-period. The results suggest that feeding of B. cereus var. toyoi to sows may result in beneficial effects on piglet health status independent of their feed supplementation.     

Influence of the probiotic strain Bacillus cereus var. toyoi on the development of enterobacterial growth and on selected parameters of bacterial metabolism in digesta samples of piglets

The effect of the probiotic strain Bacillus cereus var . toyoi on enterobacterial growth and selected metabolic parameters in digesta samples from piglets before and shortly after weaning was studied. Growth capacities of enterobacteria in digesta samples were significantly reduced, but no correlation was found between enterobacterial growth and concentrations of short-chain fatty acids and lactic acid, which are often influenced by other probiotic preparations. Tendencies for higher amounts of short-chain fatty acids were only recorded in digesta samples from the end of the jejunum, caecum and colon from piglets receiving probiotic-supplemented feed. Lactic acid concentrations were reduced by the presence of B. cereus var. toyoi in feed in the first segment of the jejunum during the suckling period, while samples of the lower parts displayed an increased amount 4 days after weaning. Microbial activity of taurocholine deconjugating enzymes was significantly reduced in jejunum digesta from probiotic-fed piglets. There are strong indications that the early uptake of already modified microbial populations from faeces of the mother sow aids modification of intestinal microbial communities and their metabolic activities in piglets.     

Effects of probiotic Bacillus cereus toyoi on abdominal fat accumulation in the Japanese quail (Coturnix japonica)

Effects of a commercial probiotic, Bacillus cereus toyoi on abdominal fat accumulation in the Japanese quail fed either a commercial control diet (crude protein (CP), 23.5%; metabolizable energy (ME), 11.7MJ/kg) or a high‐energy diet (CP, 23.0%; ME, 13.8MJ/kg) were investigated. Four‐week‐old male birds (n = 108) were divided into four diet groups: control diet, control with probiotic, high‐energy diet and high‐energy diet with probiotic. At 8 weeks and 12 weeks of age (4 and 8 weeks of probiotic supplementation period, respectively), abdominal fat, muscles and the liver were weighed. The weight of total fat and protein in the muscles and liver were also determined. A rectal temperature was recorded weekly. Bodyweight and feed intake in both diets were not affected by probiotic supplementation. At 8 weeks of age, birds fed the control diet with probiotic had significantly less abdominal fat than those fed without the probiotic, and a similar tendency was seen for both diet groups at 12 weeks of age. The decrease in abdominal fat coincided with an increase in rectal temperature. These results suggest that greater metabolizable energy was consumed through elevation of heat production by supplementation of B. cereus toyoi, with hardly any energy stored as body fat. Probiotic supplementation appears to induce greater muscle weight, higher protein and lower lipid content in muscles by 8 weeks of age, and lower lipid content in muscles and greater liver weight by 12 weeks of age, in the Japanese quail.     

Bacillus cereus var. toyoii and Saccharomyces boulardii increased feed efficiency in broilers infected with Salmonella enteritidis

1. The effect on feed efficiency of two probiotics, one prepared with Saccharomyces boulardii and the other with Bacillus cereus var. toyoii, was tested in broilers infected with Salmonella enteritidis. 2. One-day-old chicks were divided at random into three groups and fed commercial feed devoid of antibiotics: group 1 was fed with non-supplemented feed, group 2 was supplemented with S. boulardii and group 3 with B. cereus. At 14 d of age the animals were challenged by the oral route with 1 x 10(7) viable S. enteritidis. 3. At d 47, average live weights were: group 1, 1.77 kg, group 2, 1.89 kg and group 3, 2.06 kg, and were significantly different. Feed conversion rates were 2.61 for group 1, 2.35 for group 2 and 2.30 for group 3. 4. We conclude that both probiotics improved feed efficiency in broilers.     

Effects of dietary Bacillus coagulans and yeast hydrolysate supplementation on growth performance,immune response and intestinal barrier function in weaned piglets

The present study investigated the effects of Bacillus coagulans and yeast hydrolysate supplementation on growth performance, immune response and intestinal barrier function of weaned piglets. Twenty-four weaned piglets with an average body weight (BW) of 6.89 ± 0.15 kg were divided into four diets for 28 days. The treatments were basal diet (control), basal diet supplemented with antibiotic (20 mg/kg colistin sulphate and 40 mg/kg bacitracin zinc, AT), probiotics (400 mg/kg Bacillus coagulans ≥5 × 109 CFU/g, BC) or yeast hydrolysate (5000 mg/kg yeast hydrolysate, YH). Average daily gain (ADG) and average daily feed intake (ADFI) were improved by AT and YH diets (p < 0.05), while BC diet only increased ADG (p < 0.05). The complement 3 (C3), lysozyme (LZM) and tumour necrosis factor-α (TNF-α) concentrations in serum were increased in BC diet (p < 0.05). Feeding AT and YH caused the increase of jejunal villus height (p < 0.05), and a higher ratio of villus height/crypt depth was observed in AT, BC and YH groups (p < 0.05). The mRNA expression of zonula occludens-1 (ZO-1) in jejunal mucosa was up-regulated by AT, BC and YH diets (p < 0.05). Dietary AT, BC or YH inclusion decreased the interleukin-1β (IL-1β) concentration and TNF-α mRNA expression (p < 0.05), and YH supplementation even down-regulated toll-like receptor 4 (TLR4) and CD14 expressions (p < 0.05). In summary, the dietary administration of BC or YH both improves growth performance through promoting the intestinal barrier function, indicating both of them can serve as potential alternatives to antibiotics growth promoters for the piglet production.     

Effect of CLA on performance and immune response of weanling piglets

An experiment was conducted to evaluate the effect of CLA on the immune response and performance of piglets when subjected to an immune challenge. A total of 32 weanling pigs (17 to 23 d of age) with an initial BW of 8.9 kg were allotted to a 3 × 2 factorial arrangement of treatments. There were 3 levels of dietary CLA (0%, 1%, and 2%) and 2 levels of lipopolysaccharide (LPS) challenge (unchallenged and challenged). Challenged pigs were challenged on d 7 and 21. On d 4 and 18, all pigs were inoculated with BSA for assessment of IgG production. There was no difference in growth performance among piglets receiving different CLA supplementation levels. However, LPS-challenged piglets had poorer BW (P < 0.05), ADFI (P < 0.01), and ADG (P < 0.001) compared with the control group at d 35 postweaning. Lipopolysaccharide-challenged piglets also had increased respiratory rate (P < 0.001) and rectal temperature (P < 0.001), and decreased plasma proteins, hematocrit, and white blood cell counts (P < 0.05). Production of IgG against BSA was increased in the 1% CLA supplementation group (P < 0.001), indicating that CLA has an immunomodulatory effect. Supplementation with CLA did not affect lymphocyte proliferation, percentage of CD4(+) and CD8(+) cells, plasma proteins, red and white blood cell count, respiratory rate, or rectal temperature after LPS challenge. Although CLA supplementation did not influence growth performance or certain immune system measurements, the increased IgG titers with 1% CLA dietary supplementation indicate that it has a beneficial effect on the humoral immune system of weaned piglets.     

芽孢杆菌和果寡糖在仔猪营养中的应用

随着全球性生态环保意识的增强和人们对食品安全的关注 ,新型环保饲料添加剂的研究开发与应用成为饲料科技工作者迫在眉睫的任务。微生态理论 (康白 ,2 0 0 2 )认为 ,微生态失调可引起病理状态或疾病 ,而病理状态和疾病又会使微生态失调进一步恶化。去除引起微生态失调的因素 ,     

Effect of benzoic acid and combination of benzoic acid with a probiotic containing Bacillus cereus var. Toyoi in weaned pig nutrition

The purpose of this study was to assess the efficacy of a probiotic containing Bacillus cereus var. Toyoi spores (Toyocerin) and benzoic acid (VevoVitall) on growth performance and diarrhoea in weaning pigs, against negative controls. The trial groups were as follows: (a) NC group (Negative Controls): No treatment (b) TOYO group: Same feed as in the controls plus Toyocerin at a dose of 1 x 10(9) Bacillus cereus var. Toyoi spores/kg feed, (c) BA group: Same feed as in the controls plus VevoVitall at a dose of 5 g/kg feed (5000 ppm benzoic acid) and (d) TOYO+BA group: Same feed as in the controls plus Toyocerin at a dose of 1 x 10(9) Bacillus cereus var. Toyoi spores and VevoVitall at a dose of 5 g/kg feed. In conclusion, the results of this study indicated that administration of Bacillus cereus var. Toyoi spores at 1 x 10(9)/kg feed or benzoic acid at a dose of 5000 ppm or the combination of 1 x 10(9) Bacillus cereus var. Toyoi spores and 5000 ppm of benzoic acid/kg feed, improved the growth performance parameters and reduced the severity of diarrhoea in weaning pigs.     

Bacillus cereus from the environment is genetically related to the highly pathogenic B. cereus in Zambia

To follow-up anthrax in Zambia since the outbreak in 2011, we have collected samples from the environment and the carcasses of anthrax-suspected animals, and have tried to isolate Bacillus anthracis. In the process of identification of B. anthracis, we collected two isolates, of which colonies were similar to B. anthracis; however, from the results of identification using the molecular-based methods, two isolates were genetically related to the highly pathogenic B. cereus, of which clinical manifestation is severe and fatal (e.g., pneumonia). In this study, we showed the existence of bacteria suspected to be highly pathogenic B. cereus in Zambia, indicating the possibility of an outbreak caused by highly pathogenic B. cereus.     

Cellular immune response and immunotoxicity induced by DON (deoxynivalenol) in piglets

Veterinary Research Communications -     

Maternal immunity enhances systemic recall immune responses upon oral immunization of piglets with F4 fimbriae

F4 enterotoxigenic Escherichia coli (ETEC) cause diarrhoea and mortality in piglets leading to severe economic losses. Oral immunization of piglets with F4 fimbriae induces a protective intestinal immune response evidenced by an F4-specific serum and intestinal IgA response. However, successful oral immunization of pigs with F4 fimbriae in the presence of maternal immunity has not been demonstrated yet. In the present study we aimed to evaluate the effect of maternal immunity on the induction of a systemic immune response upon oral immunization of piglets. Whereas F4-specific IgG and IgA could be induced by oral immunization of pigs without maternal antibodies and by intramuscular immunization of pigs with maternal antibodies, no such response was seen in the orally immunized animals with maternal antibodies. Since maternal antibodies can mask an antibody response, we also looked by ELIspot assays for circulating F4-specific antibody secreting cells (ASCs). Enumerating the F4-specific ASCs within the circulating peripheral blood mononuclear cells, and the number of F4-specific IgA ASCs within the circulating IgA⁺ B-cells revealed an F4-specific immune response in the orally immunized animals with maternal antibodies. Interestingly, results suggest a more robust IgA booster response by oral immunization of pigs with than without maternal antibodies. These results demonstrate that oral immunization of piglets with F4-specific maternal antibodies is feasible and that these maternal antibodies seem to enhance the secondary systemic immune response. Furthermore, our ELIspot assay on enriched IgA⁺ B-cells could be used as a screening procedure to optimize mucosal immunization protocols in pigs with maternal immunity.     

Purification and some properties of a Bacillus cereus mouse lethal toxin.

A mouse lethal toxin (MLT) produced by Bacillus cereus isolated in vomiting-type food poisoning was purified by chromatography on DEAE-Sephadex A-25 followed by gel filtration on Sephadex G-75. Purified MLT possessed a molecular weight of 33,000-34,000. It showed mouse lethality and hemolytic (HL) activity on sheep and rabbit erythrocytes; the latter erythrocytes were more weakly hemolyzed than the former ones. However, fluid accumulation in mouse ligated intestinal loops was not induced by purified MLT at the highest concentration used. Both MLT and HL activities were stable at pH 6-9, during storage at -20 degrees C for 8 weeks, and resistant to papain, cholesterol, lecithin, and dithiothreitol treatments. Most activity was lost during storage at 4 degrees C or 25 degrees C for 2 weeks or upon treatment with trypsin, trypanblue, or ethanol. The activities were resistant to heating at 37 degrees C for 5 min, less resistant at 98 degrees C for 5 min, and sensitive at 60 degrees C for 5 min. It can be concluded from the results that MLT is different from the diarrheagenic toxin produced by B. cereus isolated in diarrheal-type food poisoning, but is similar to, if not identical, hemolysin II.