Characterization of the Effects of a Novel Probiotic on Salmonella Colonization of a Piglet-Derived Intestinal Microbiota Using Improved Bioreactor.

The carriage of Salmonella in pigs is a major concern for the agri-food industry and for global healthcare systems. Humans could develop salmonellosis when consuming contaminated pig products. On the other hand, some Salmonella serotypes could cause disease in swine, leading to economic losses on farms. The purpose of the present study was to characterize the anti-Salmonella activity of a novel Bacillus-based probiotic using a bioreactor containing a piglet-derived intestinal microbiota. Two methods of probiotic administration were tested: a single daily and a continuous dose. Salmonella enumeration was performed using selective agar at T24h, T48h, T72h, T96h and T120h. The DNA was extracted from bioreactor samples to perform microbiome profiling by targeted 16S rRNA gene sequencing on Illumina Miseq. The quantification of short-chain fatty acids (SCFAs) was also assessed at T120h. The probiotic decreased Salmonella counts at T96 for the daily dose and at T120 for the continuous one. Both probiotic doses affected the alpha and beta diversity of the piglet-derived microbiota (p < 0.05). A decrease in acetate concentration and an increase in propionate proportion were observed in the continuous condition. In conclusion, the tested Bacillus-based product showed a potential to modulate microbiota and reduce Salmonella colonization in a piglet-derived intestinal microbiota and could therefore be used in vivo.

A short-term bioreactor assay to assess the effect of essential oils on a microbiota derived from piglet's intestinal content.

BACKGROUND: Modulating the microbiota is an emerging way to improve pig health. In-vitro bioreactor systems can be used to reproduce intestinal microbiota to study modulating avenues. In this study, a continuous feeding system to support a microbiota derived from piglet colonic contents, over 72 h, was developed. The microbiota from piglets was collected and used as inoculum. The culture media was derived from an artificial digestion of piglet feed. The microbiota diversity in time, the reproducibility between replicates and the diversity of the bioreactor microbiota compared to the inoculum was assessed. Essential oils were used as a proof of concept to assess the in vitro microbiota modulation. The microbiota diversity was assessed by 16S rRNA amplicon sequencing. Quantitative PCR was also used for total bacteria, lactobacilli and Enterobacteria. RESULTS: At the start of the assay, the bioreactor microbiota diversity was similar to the inoculum. Time and replication affected the bioreactor microbiota diversity. Between 48 and 72 h, no statistical variation of the microbiota diversity was observable. After a 48 h running period, thymol and carvacrol were added at 200 ppm or 1000 ppm for 24 h. No microbiota modification was observed by sequencing. Quantitative PCR results showed a significant growth of lactobacilli when thymol was used at 1000 ppm, where only a trend was observed with the 16S analysis. CONCLUSIONS: This study presents a bioreactor assay that can be used as a tool for rapid screening of additives and suggests that the effects of essential oils on the microbiota are subtle, acting against a few bacterial genera.

Effect of the carrier material, drying technology and dissolution media on the viability of Lactobacillus fermentum K73 during simulated gastrointestinal transit.

The goal of this study was to determine the effect of the carrier material, drying technology and dissolution media during the passage of L. fermentum K73 through a dynamic in vitro digestion system (IViDiS). The carrier materials were (i) culture medium with growing micro-organisms and (ii) culture medium with maltodextrin : sweet whey [0.6 : 0.4]. The carrier materials were dried by spray-drying and freeze-drying to obtain four types of powders. The dissolution media consisted of water and 1% fat milk. The powders were tested using an in vitro dynamic digestion system (IViDiS). The results showed that powders derived from culture medium had the highest protective effect on the viability of L. fermentum K73 in both dissolution media and that survival increased when the powders were tested in milk. The modified Gompertz model was used to model L. fermentum K73 behaviour during the digestion process. The model showed that cells entrapped in culture medium had the longest lag phase and the slowest inactivation rate when evaluated in milk.

Polyphasic characterization of the lactic acid bacteria in kefir.

The lactic acid bacteria of kefir were isolated and characterized using phenotypical, biochemical, and genotypical methods. Polyphasic analyses of results permitted the identification of the microflora to the strain level. The genus Lactobacillus was represented by the species Lb. kefir and Lb. kefiranofaciens. Both subspecies of Lactococcus lactis (lactis and cremoris) were isolated. Leuconostoc mesenteroides subsp. cremoris was also found. The kefir studied contained few species of lactic acid bacteria but showed a high number of different strains. We found that the polyphasic analysis approach increases the confidence in strain determination. It helped confirm strain groupings and it showed that it could have an impact on the phylogeny of the strains.