Effects of the probiotic Enterococcus faecium NCIMB 10415 on selected lactic acid bacteria and enterobacteria in co-culture.

Enterococcus faecium NCIMB 10415 is used as a probiotic for piglets and has been shown to modify the porcine intestinal microbiota. However, the mode of action of this probiotic modification is still unclear. One possible explanation is the direct growth inhibiting or stimulating effect of the probiotic on other indigenous bacteria. Therefore, the aim of the present study was to examine the growth interactions of the probiotic with different indigenous porcine bacteria in vitro. Reference strains were cultivated with the probiotic E. faecium strain NCIMB10415 (SF68) in a checkerboard assay with 102 to 105 cells/ml inoculum per strain. Growth kinetics were recorded for 8 h and used to determine specific growth of the co-cultures. Additionally, total DNA was extracted from the co-cultures at the end of the incubation to verify which strain in the co-culture was affected. Co-cultivation with eight Enterococcus spp. tester strains showed strain-specific growth differences. Three of four E. faecium strains were not influenced by the probiotic strain. PCR results showed reduced growth of the probiotic strain in co-culture with E. faecium DSM 6177. Three of four Enterococcus faecalis strains showed reduced specific growth in co-culture with the probiotic strain. However, E. faecalis DSM 20478 impaired growth of the probiotic E. faecium strain. The growth of Lactobacillus johnsonii DSM 10533 and Lactobacillus reuteri DSM 20016 was enhanced in co-culture with the probiotic strain, but co-cultivations with Lactobacillus mucosae DSM13345 or Lactobacillus amylovorus DSM10533 showed no differences. Co-cultures with the probiotic E. faecium showed no impact on the growth rate of four different enterobacterial reference strains (2 strains of Salmonella enterica and 2 strains of Escherichia coli), but PCR results showed reduced cell numbers for a pathogenic E. coli isolate at higher concentration of the probiotic strain. As the in vitro effect of the probiotic E. faecium on enterococci was strain specific and the growth of certain Lactobacillus spp. was enhanced by the probiotic, these results indicate a direct effect of the probiotic on certain members of the porcine gastro intestinal microbiota.

Cross-talk Between Host, Microbiome and Probiotics: A Systems Biology Approach for Analyzing the Effects of Probiotic Enterococcus faecium NCIMB 10415 in Piglets.

A comprehensive data-set from a multidisciplinary feeding experiment with the probiotic Enterococcus faecium was analyzed to elucidate effects of the probiotic on growing piglets. Sixty-two piglets were randomly assigned to a control (no probiotic treatment) and a treatment group (E. faecium supplementation). Piglets were weaned at 26 d. Age-matched piglets were sacrificed for the collection of tissue samples at 12, 26, 34 and 54 d. In addition to zootechnical data, the composition and activity of intestinal microbiota, immune cell types, and intestinal responses were determined. Our systems analysis revealed clear effects on several measured variables in 26 and 34 days old animals, while response patterns varied between piglets from different age groups. Correlation analyses identified reduced associations between intestinal microbial communities and immune system reactions in the probiotic group. In conclusion, the developed model is useful for comparative analyses to unravel systems effects of dietary components and their time resolution. The model identified that effects of E. faecium supplementation most prominently affected the interplay between intestinal microbiota and the intestinal immune system. These effects, as well as effects in other subsystems, clustered around weaning, which is the age where piglets are most prone to diarrhea.

Individual responses of mother sows to a probiotic Enterococcus faecium strain lead to different microbiota composition in their offspring.

Pregnant gilts were fed the probiotic Enterococcus faecium NCIMB10415 (SF68) one month before birth of piglets. DNA extracts of sow faeces taken in weekly intervals as well as extracts from the intestine of their offspring during the suckling period at 12 and 26 days of life were analysed by denaturing gradient gel electrophoresis (DGGE) and quantitative PCR. DGGE profiles of faecal bacterial communities from three out of six probiotic-fed sows were distinctly different from the control and other probiotic-fed sows at all time points after probiotic supplementation. The probiotic-fed sows and their offspring were therefore divided into non-responder (n=3) and responder (n=3) groups. The probiotic strain significantly increased faecal lactobacilli cell numbers in mother sows, which could be assigned to a significant increase of Lactobacillus amylovorus and Lactobacillus acidophilus. Responding sows showed a more pronounced increase than non-responding sows. Similarly, suckling piglets from non-responding and responding sows showed numeric and significant differences for different bacterial groups and species. DGGE profiles of suckling piglets from responding sows also grouped more closely than profiles from control animals. Non-metric multiscaling of suckling piglets showed the same tendency for suckling piglets, but not for post-weaning piglets. This study showed that the probiotic E. faecium strain modified the faecal microbiota of sows. This modification is carried over to their offspring, but leads to changes that do not mirror the quantitative composition in the mother sow. Individual variations in the bacterial composition of mother sows before probiotic feed intake may influence the impact of a probiotic in sows and their offspring.