Gerd Schatzmayr

Effects of probiotic inclusion levels in broiler nutrition on growth performance, nutrient digestibility, plasma immunoglobulins, and cecal microflora composition

The aim of this work was to investigate the effect of inclusion levels of a 5-bacterial species probiotic in broiler nutrition. Five hundred twenty-five 1-d-old male Cobb broilers were allocated in 5 experimental treatments for 6 wk. The experimental treatments received a corn-soybean coccidiostat-free basal diet and depending on the addition were labeled as follows: no addition (C), 10(8) cfu probiotic/kg of diet (P1), 10(9) cfu probiotic/kg of diet (P2), 10(10) cfu probiotic/kg of diet (P3), and 2.5 mg of avilamycin/kg of diet (A). Each treatment had 3 replicates of 35 broilers each. Treatment effects on broiler growth performance and biomarkers such as ileal and total tract nutrient digestibility, plasma Ig concentration, and cecal microflora composition were determined. Differences among treatments were considered significant when P < or = 0.05. Overall BW gain was significantly higher in treatment P1 (2,293 g) compared with P2 (2,163 g), C (2,165 g), and P3 (2,167 g), with A (2,230 g) being intermediate and not different from P1. Overall feed conversion ratio values were similar and significantly better for P1 (1.80) and A (1.80) compared with P2 (1.87), C (1.89), and P3 (1.92). Ileal apparent digestibility coefficients (ADC) of CP and ether extract were higher in A. Generally, treatments A and P1 showed an improved total tract ADC for DM, organic matter, ash, ether extract, and AME(n) values. The total tract ADC of CP was higher in P1, C, and P2. There were no differences between treatments regarding plasma Ig in 14- and 42-d-old broilers. Treatments P2 and P3 were effective at beneficially modulating cecal microflora composition. In particular, the lower cecal coliform concentration (log cfu/g of wet digesta) was seen in P2 (6.12) and P3 (4.90) in 14- and 42-d-old broilers, respectively, whereas at 42 d, P3 and P2 had the highest Bifidobacterium (8.31; 8.08) and Lactobacillus concentrations (8.20; 7.86), respectively. It is concluded that probiotic inclusion level had a significant effect on broiler growth responses, nutrient ADC, AME(n), and cecal microflora composition.

In vitro antagonistic activities of animal intestinal strains against swine-associated pathogens.

A wide range of enteropathogens cause costly diarrhoeal diseases in fattening piglets and account for food-related infections in humans. The objective of this study was to screen beneficial bacterial strains from the gastrointestinal tract of various animal sources for antagonistic activity against diverse pathogens associated with hazardous pig production times. Using agar spot assays, 15 well-characterized strains belonging to Lactobacillus, Enterococcus, Bifidobacterium and Bacillus were studied for inhibition of Clostridium perfringens type A, various serovars of enterotoxigenic Escherichia coli and Salmonella enterica, as well as Brachyspira pilosicoli. Strong antagonists were further analyzed by studying their cell-free supernatants with and without pH neutralization, proteinase K and catalase treatment. Enterobacteriaceae were effectively inhibited by Lactobacillus salivarius and Lactobacillus reuteri strains, independent from the animal source, and on a lower level by single strains belonging to Lactobacillus mucosae, Lactobacillus amylovorus and Bifidobacterium thermophilum, due to organic acid production. The Bacillus subtilis strain was found to produce an anti-clostridial and anti-Brachyspira metabolite of proteinaceous nature. Homofermentative lactobacilli and B. thermophilum could suppress the growth of B. pilosicoli, the causative agent of intestinal spirochaetosis, whereas heterofermentative strains belonging to L. reuteri and L. mucosae had no effect. The lactic acid bacteria exerted their activity primarily by organic acid release, except one Enterococcus faecium and L. amylovorus strain, which exhibited antagonism through joint activity of lactate and hydrogen peroxide. The findings of this study provide a basis for further in vitro studies and encourage feeding studies to evaluate the antagonistic potential of promising strains in pig production.

Investigation of different yeast strains for the detoxification of ochratoxin A

High concentrations of ochratoxin A (OTA) in feed lead to growth depression in animals. It has been reported that binders can be used for deactivating aflatoxins but not for other mycotoxins without negatively influencing the animals health. In this study a strain from the genus ofTrichosporon with the ability to cleave ochratoxin A very selectively into phenylalanine and the non-toxic ochratoxin α (OTα) could be isolated. This strain was selected from a pool of OTA detoxifying microorganism by carrying out several investigations.Trichosporon sp. nov. can be fermented and stabilized. In a feeding trial with broilers lyophilizedTrichosporon-cells could compensate performance losses caused by OTA.

Capability of yeast derivatives to adhere enteropathogenic bacteria and to modulate cells of the innate immune system.

Yeast derivatives including yeast cell wall components are promising alternatives to antibiotics with respect to the promotion of health and performance in livestock, based on their capacity to bind enteropathogenic bacteria and to beneficially modulate the immune system. However, these mode(s) of action both in vitro and in vivo are still not well understood. Furthermore, standardization and reproducibility of in vitro techniques (microbiology, cell culture assays) are critical features for the application of yeast derivatives as well as for the proof of effectiveness. Yeast cell wall products are suggested as anti-adhesive agents and are thus proposed to prevent attachment of certain intestinal bacteria by providing alternative adhesion sites to enterobacteria, which contain mannose-specific type I fimbriae such as Escherichia coli or Salmonella spp. and which is well documented. Various in vitro assay techniques have become of paramount importance for biotechnological research since they allow for determination and quantification of potential mode(s) of action. However, in vitro assays may be criticized by product end users as not accurately reflecting in vivo responses. Pro and cons of different assays and their bias will be discussed specifically regarding yeast cell wall components and adhesion of enteropathogenic bacteria. Immunomodulation is a therapeutic approach intervening in auto-regulating processes of the defense system. Yeast derivatives such as beta-glucans are proposed to interact with cells of the innate immune system by receptor recognition. Controversial data in literature and mode(s) of action are reviewed and discussed here.

Net effect of an acute phase response—Partial alleviation with probiotic supplementation

The acute phase response (APR) is characterized by inflammation, fever, and altered organ metabolism resulting in muscle catabolism and anorexia. Lipopolysaccharide (LPS)-induced APR may reflect depressed growth and appetite loss. Therefore, a 1-wk growth experiment was conducted to examine whether dietary supplementation of a multispecies probiotic (PoultryStar) would alleviate growth suppression and anorexia caused by LPS-induced APR. The experiment was designed with 4 treatments (n = 8 cages/treatment; 6 birds/cage) starting at 14 d of age. Before (0 to 14 d of age) and for the experiment (14 to 21 d of age), male broiler chicks were fed diets devoid of probiotic or were supplemented with 1.7 x 10(8) cfu/kg of probiotic. At 14 d of age, birds fed the diet devoid of probiotic were further divided into 3 treatments: an unchallenged positive control, LPS-challenged negative control (LPS-NC), and a treatment that was pair-fed to LPS-NC. The probiotic-fed birds were also then challenged with LPS. The LPS (Escherichia coli 055:B5) was injected intraperitoneally 4 times at 48-h intervals at 1 mg/kg of BW. The LPS challenge dramatically depressed BW gain from 14 to 21 d of age by 22% (P < 0.001). However, 41% of growth depression was attributable to factors other than feed intake reduction when compared with the pair-fed treatment. Probiotic supplementation recovered 17% of depressed growth (vs. LPS-NC; P = 0.068), but this improved growth was not due to improvements in feed intake (P = 0.47). However, recovery of feed intake of the probiotic + LPS birds occurred 48-h earlier than the LPS-NC birds. Growth depression induced by LPS administration resulted in an overall relative feed intake (vs. positive control) of 0.83 and also decreased net energy and protein accretion. Probiotic supplementation did not alleviate the reduction in net energy or protein accretion induced by LPS. In conclusion, APR (induced by LPS administration) diverted a large portion of consumed nutrients from tissue accretion. Probiotic supplementation lessened the anorexic effects of LPS resulting in a trend toward BW gain improvement versus the LPS-NC.

Quantitative in vitro assay to evaluate the capability of yeast cell wall fractions from Trichosporon mycotoxinivorans to selectively bind gram negative pathogens

Yeast cell wall fractions have been proposed to bind enteropathogenic bacteria. The aim of this study was to develop a quantitative assay by measuring the optical density as growth parameter of adhering bacteria. The exponential growth phase of adhering bacteria was determined by optical density reading and compared with the colony count (CFU/mL). A linear regression was compiled and the bacterial number bound to the yeast cell wall product could be determined. Further focus was the investigation of a yeast cell wall from strain Trichosporon mycotoxinivorans (MTV) for its ability to bind gram negative Salmonella, E. coli and Campylobacter strains and gram positive probiotic bacteria of the genera lactobacilli and bifidobacteria as well as gram positive Clostridium perfringens quantitatively. The gram negative probiotic strain E. coli Nissle 1917 was also investigated. Seven out of 10 S. Typhimurium and S. Enteritidis strains adhered to the cell wall product with an amount between 10(3) and 10(4) CFU/10 μg. Four out of 7 E. coli strains showed an average binding capability (10(2) CFU/10 µg) whereas 4×10(3) E. coli F4 cells bound per 10 μg yeast cell wall. E. coli 0149 K91, E. coli 0147 K89, C. jejuni and C. perfringens as well the genera lactobacilli and bifidobacteria did not bind to the yeast cell wall. E. coli Nissle 1917 was bound with 2×10(2) CFU/10 μg. These results demonstrate that cell wall from MTV can be used to differentially bind E. coli spp. and Salmonella spp. up to 8×10(4) CFU/10 μg. Thus certain yeast cell walls may prevent enteric infections caused by selective bacteria. This methodical approach would be an accurate tool in the feed industry for quality control of yeast cell wall products.

Identification and antimicrobial susceptibility of porcine bacteria that inhibit the growth of Brachyspira hyodysenteriae in vitro.

Aims:u2002 To identify bacilli, lactic acid bacteria and bifidobacteria that inhibit the growth of Brachyspira hyodysenteriae.

Comparative evaluation of probiotic and salinomycin effects on performance and coccidiosis control in broiler chickens

The annual financial loss to the poultry industry as a result of coccidiosis has been estimated at about US

Effect of dietary inclusion level of a multi-species probiotic on broiler performance and two biomarkers of their caecal ecology

3 billion. The objective of this study was to evaluate and compare the effects of probiotics and salinomycin as feed additives on performance and coccidiosis control in male broilers raised to 42 d of age. The study consisted of 360 Cobb male broiler chickens randomly allocated to 4 groups each with 3 replicates. Group 1: untreated, unchallenged negative control group (NC); group 2: untreated, challenged positive control group (PC); group 3: negative control supplemented with salinomycin 66 mg/kg, challenged group (Sal); and group 4: negative control supplemented with probiotics, challenged (Prob mix). On d 15, all birds (except group 1) were challenged with approximately 75,000, 25,000, and 75,000 of Eimeria acervulina, Eimeria maxima, and Eimeria tenella oocytes, respectively, that were mixed into the feed. Feed conversion ratio and mortality were recorded throughout the experiment. On d 21 and 42, intestinal lesions and litter conditions were scored. On d 7, 14, 21, 28, 35, and 42, oocyst counts were determined from 10 freshly collected fecal samples per pen. The results showed that mortality, litter, and lesion scores at d 21 and 42, and oocyst shedding at d 21 did not differ significantly between the Prob mix and the Sal groups. However on d 28, oocyst shedding was significantly lower in the Sal group than in the PC group but insignificantly lower than the Prob mix group. Body weights of the Prob mix group at d 42 were significantly lower than the Sal group; however, the feed conversion ratio values were similar between the 2 groups. The results of this study showed that probiotics supplementation could be considered as a potential strategy to control coccidiosis in broiler chickens.

Evidence of ochratoxin A-detoxification activity of rumen fluid, intestinal fluid and soil samples as well as isolation of relevant microorganisms from these environments.

The effect of the dietary inclusion level of a three-species probiotic on broiler performance, nutrient digestibility, caecal microbiota composition and volatile fatty acid (VFA) pattern was evaluated. Day-old Cobb broilers (n = 448) were allocated in four treatments for 6 weeks. Each treatment had four replicates (two per gender) of 28 broilers each. Depending on the type of addition per kg basal diet, treatments were C (no other addition), PL (108 colony forming units of probiotic), PH (109 colony forming units of probiotic) and A (2.5 mg avilamycin). Overall bodyweight gain was better (P = 0.002) in PL and PH than in the control (2082 g) by 8.7% and 7.5%, respectively, while treatment PL did not differ from A (2341 g), which showed the highest bodyweight gain. The ileal and total-tract apparent digestibility of DM and the apparent metabolisable energy content corrected for N improved linearly (P ≤ 0.05) with the probiotic level. Fluorescent in situ hybridisation analysis showed caecal Bifidobacterium levels to increase linearly (P = 0.006) with the probiotic level. Probiotic administration resulted in altered caecal VFA patterns compared with the control. Gender effects (P ≤ 0.05) were noted for caecal levels of C. histolyticum group, Bacteroides fragilis group and Streptococcus spp., while interactions (P ≤ 0.05) of treatment with gender were seen for Bifidobacterium and all VFA components, except for acetate. In conclusion, beneficial effects on bodyweight gain, DM digestibility, apparent metabolisable energy content corrected for N, caecal Bifidobacterium levels and VFA patterns were noted with both probiotic inclusion levels.