Assessing the Impact of Flavophospholipol and Virginiamycin Supplementation on the Broiler Microbiota: a Prospective Controlled Intervention Study.

The antibiotic growth promoters (AGPs) flavophospholipol and virginiamycin have been widely used for decades in food animal production. AGP activity is believed to be partly modulated by gut microbial composition although exact AGP-induced changes remain unclear. In a controlled intervention study, we studied the effect of flavophospholipol and virginiamycin on the broiler chicken ileal microbiota spanning from birth to 39 days. Using 16S rRNA gene profiling and prediction of metabolic activity, we show that both AGPs result in dynamic microbial shifts that potentially increase anti-inflammatory mechanisms and bioavailability of several essential nutrients by decreasing degradation (flavophospholipol) or increasing biosynthesis (virginiamycin). Further, virginiamycin-supplemented broilers showed increased colonization with potentially pathogenic bacteria, Clostridium perfringens, Campylobacter, and Escherichia/Shigella spp. Overall, we show that both AGPs induce microbial changes potentially beneficial for growth. However, the increase in (foodborne) pathogens shown here with virginiamycin use could impact not only broiler mortality but also human health. IMPORTANCE Antibiotic growth promoters (AGPs) are commonly used within poultry farming to increase muscle growth. Microbial composition in the gut is known to be influenced by AGP use although exact AGP-induced changes remain unclear. Utilizing 16S rRNA gene profiling, this study provides a first head-to-head comparison of the effect of the two most commonly used AGPs, flavophospholipol and virginiamycin, on the broiler chicken ileum microbiota over time. We found that supplementation with both AGPs altered ileal microbial composition, thereby increasing potential bioavailability of essential nutrients and weight gain. Flavophospholipol showed a slight benefit over virginiamycin as the latter resulted in more extensive microbial perturbations including increased colonization by enteropathogens, which could impact broiler mortality.

Combination of Bacillus licheniformis and Salinomycin: Effect on the Growth Performance and GIT Microbial Populations of Broiler Chickens.

The aim of the study was to investigate the effect of Bacillus licheniformis and salinomycin supplementation in broiler diets as individual factors or in combination on the growth performance, GIT morphometry, and microbiota populations. Four hundred one-day-old Ross 308 chicks were randomly distributed to four dietary treatments (10 replicates, 10 birds each). The following treatments were applied: NC-no additives; NC + SAL-salinomycin addition (60 mg/kg diet), NC + PRO-B. licheniformis DSM 28710 preparation (1.6 × 109 CFU/kg; 500 mg/kg diet), and NC + SAL + PRO-combination of salinomycin and B. licheniformis. Probiotic administration resulted in improvement (p < 0.05) of the performance parameters, including body weight gain (1-10 d, and 11-22 d) and feed conversion ratio (11-22 d, 1-36 d). An interaction (p < 0.05) between experimental factors was observed in terms of lower pH values in the crop (tendency, p = 0.053) and ceca. Both factors lowered the alpha diversity and Enterobacteriaceae and promoted Bacillaceae communities in the jejunum (p < 0.05). Interactions were also observed in terms of reducing Clostridiaceae in the ceca. In conclusion, the combined use of B. licheniformis and salinomycin in broilers' diets had beneficial effects.

In-feed bambermycin medication induces anti-inflammatory effects and prevents parietal cell loss without influencing Helicobacter suis colonization in the stomach of mice.

The minimum inhibitory concentration of bambermycin on three porcine Helicobacter suis strains was shown to be 8 µg/mL. The effect of in-feed medication with this antibiotic on the course of a gastric infection with one of these strains, the host response and the gastric microbiota was determined in mice, as all of these parameters may be involved in gastric pathology. In H. suis infected mice which were not treated with bambermycin, an increased number of infiltrating B-cells, T-cells and macrophages in combination with a Th2 response was demonstrated, as well as a decreased parietal cell mass. Compared to this non-treated, infected group, in H. suis infected mice medicated with bambermycin, gastric H. suis colonization was not altered, but a decreased number of infiltrating T-cells, B-cells and macrophages as well as downregulated expressions of IL-1ß, IL-8M, IL-10 and IFN-γ were demonstrated and the parietal cell mass was not affected. In bambermycin treated mice that were not infected with H. suis, the number of infiltrating T-cells and expression of IL-1ß were lower than in non-infected mice that did not receive bambermycin. Gastric microbiota analysis indicated that the relative abundance of bacteria that might exert unfavorable effects on the host was decreased during bambermycin supplementation. In conclusion, bambermycin did not affect H. suis colonization, but decreased gastric inflammation and inhibited the effects of a H. suis infection on parietal cell loss. Not only direct interaction of H. suis with parietal cells, but also inflammation may play a role in death of these gastric acid producing cells.

Deoxynivalenol impairs hepatic and intestinal gene expression of selected oxidative stress, tight junction and inflammation proteins in broiler chickens, but addition of an adsorbing agent shifts the effects to the distal parts of the small intestine.

Broiler chickens are rather resistant to deoxynivalenol and thus, clinical signs are rarely seen. However, effects of subclinical concentrations of deoxynivalenol on both the intestine and the liver are less frequently studied at the molecular level. During our study, we investigated the effects of three weeks of feeding deoxynivalenol on the gut wall morphology, intestinal barrier function and inflammation in broiler chickens. In addition, oxidative stress was evaluated in both the liver and intestine. Besides, the effect of a clay-based mycotoxin adsorbing agent on these different aspects was also studied. Our results show that feeding deoxynivalenol affects the gut wall morphology both in duodenum and jejenum of broiler chickens. A qRT-PCR analysis revealed that deoxynivalenol acts in a very specific way on the intestinal barrier, since only an up-regulation in mRNA expression of claudin 5 in jejunum was observed, while no effects were seen on claudin 1, zona occludens 1 and 2. Addition of an adsorbing agent resulted in an up-regulation of all the investigated genes coding for the intestinal barrier in the ileum. Up-regulation of Toll-like receptor 4 and two markers of oxidative stress (heme-oxigenase or HMOX and xanthine oxidoreductase or XOR) were mainly seen in the jejunum and to a lesser extent in the ileum in response to deoxynivalenol, while in combination with an adsorbing agent main effect was seen in the ileum. These results suggest that an adsorbing agent may lead to higher concentrations of deoxynivalenol in the more distal parts of the small intestine. In the liver, XOR was up-regulated due to DON exposure. HMOX and HIF-1α (hypoxia-inducible factor 1α) were down-regulated due to feeding DON but also due to feeding the adsorbing agent alone or in combination with DON.

Development of a sanitary risk index for Salmonella seroprevalence in Belgian pig farms.

The aim of this study was to develop a scientifically based Sanitary Risk Index (SRI), defined as an objective measure of the Salmonella seroprevalence in a pig herd based on the risk factors being present on the farm. Therefore, an observational epidemiological study was adopted to infer risk factors for the Salmonella seroprevalence of market pigs. A total of 204 Belgian farrow-to-finish pig herds were included in this cross-sectional study. The antibody titre to Salmonella in sera was analysed by means of an enzyme-linked immunosorbent assay (ELISA) for an average of 58 finisher pigs on each farm. A detailed questionnaire, covering an extensive range of potential risk factors was completed by each participating pig producer. Pearson correlation coefficients between the average sample to positive ratio (S/P)-value of a herd and the within-herd proportion of seropositive pigs were high. Significant risk factors associated with the average S/P-value of a herd were identified by a general linear mixed model. Feeding of meal, providing wet feed, having a hygienic-lock facility, using boot baths, applying the strict all in/all out procedure, programming the temperature in the zone of thermal neutrality and disinfecting between batches were all associated with lower average S/P-values. Sampling in summer, using a clean downtime, decreasing floor space per animal as well as increasing herd size were related with higher average S/P-values. The SRI consists of the above-specified risk factors together with their relative weight. Determining the Salmonella risk of a new herd by the SRI is primarily based on the quantification of the farm specific risk factors present and results in an average S/P-value of the herd. The model was validated using a set of conventional farms. In conclusion, the SRI is a useful preliminary screening tool which forms the basis for targeted sampling but cannot replace the serological herd classification with regard to Salmonella prevalence.

Discrepancies between the isolation of Salmonella from mesenteric lymph nodes and the results of serological screening in slaughter pigs.

Most Salmonella control programmes are based on serological testing in the slaughterhouse. However, from a point of view of carcass contamination, it is rather the presence of Salmonella spp. in the animal at the time of slaughter that is important. The aim of this cross-sectional study was to investigate the possible discrepancies between the isolation of Salmonella spp. in the mesenteric lymph nodes and the results of serological screening. In total, 1821 fattening pigs originating from 60 Belgian farrow-to-finish herds were sampled in the slaughterhouse. The serum samples were analysed using an indirect mix-ELISA for the presence of Salmonella antibodies and evaluated at 3 cut-off values namely 10, 20, and 40% Optical Density (OD). All mesenteric lymph node samples were submitted to qualitative Salmonella isolation and a representative number of isolates was serotyped. From each herd, 30 animals were screened both serologically and bacteriologically and the herd was considered as positive when at least one sample was positive. At the herd level, 83.6% (cut-off OD 40%) to 100.0% (cut-off OD 10%) of the herds from which Salmonella had been isolated were evaluated as seropositive. At the individual level, only 34.5% (cut-off OD 40%) to 82.8% (cut-off OD 10%) of the animals from which Salmonella had been isolated were seropositive. Overall, a weak agreement was found between bacteriology and serology for Salmonella diagnosis. If pig herds are categorised using serological tests in the slaughterhouse, one should be aware of the fact that slaughter pigs can still harbour Salmonella spp. in the mesenteric lymph nodes, without being detected in serological tests. The cut-off value used to evaluate a sample as serologically positive and the number of samples per herd are of major importance to classify herds correctly in order to protect human health.