Genetic relationships among Enterococcus faecalis isolates from different sources as revealed by multilocus sequence typing.

Enterococcus faecalis is part of the natural gut flora of humans and other mammals; some isolates are also used in food production. So, it is important to evaluate the genetic diversity and phylogenetic relationships among E. faecalis isolates from different sources. Multilocus sequence typing protocol was used to compare 39 E. faecalis isolates from Chinese traditional food products (including dairy products, acidic gruel) and 4 published E. faecalis isolates from other sources including human-derived isolates employing 5 housekeeping genes (groEL, clpX, recA, rpoB, and pepC). A total of 23 unique sequence types were identified, which were grouped into 5 clonal complexes and 10 singletons. The value of standardized index of association of the alleles (IA(S)=0.1465) and network structure indicated a high frequency of intraspecies recombination across these isolates. Enterococcus faecalis lineages also exhibited clearly source-clustered distributions. The isolates from dairy source were clustered together. However, the relationship between isolates from acidic gruel and one isolate from a human source was close. The MLST scheme presented in this study provides a sharable and continuously growing sequence database enabling global comparison of strains from different sources, and will further advance our understanding of the microbial ecology of this important species.

Molecular identification and quantification of lactic acid bacteria in traditional fermented dairy foods of Russia.

Russian traditional fermented dairy foods have been consumed for thousands of years. However, little research has focused on exploiting lactic acid bacteria (LAB) resources and analyzing the LAB composition of Russian traditional fermented dairy foods. In the present study, we cultured LAB isolated from fermented mare and cow milks, sour cream, and cheese collected from Kalmykiya, Buryats, and Tuva regions of Russia. Seven lactobacillus species and the Bifidobacterium genus were quantified by quantitative PCR. The LAB counts in these samples ranged from 3.18 to 9.77 log cfu/mL (or per gram). In total, 599 LAB strains were obtained from these samples using de Man, Rogosa, and Sharpe agar and M17 agar. The identified LAB belonged to 7 genera and 30 species by 16S rRNA and murE gene sequencing and multiplex PCR assay. The predominant LAB isolates were Lactobacillus helveticus (176 strains) and Lactobacillus plantarum (63 strains), which represented 39.9% of all isolates. The quantitative PCR results revealed that counts of 7 lactobacilli species and Bifidobacterium spp. of 30 fermented cow milk samples ranged from 1.19±0.34 (Lactobacillus helveticus in Tuva) to 8.09±0.71 (Lactobacillus acidophilus in Kalmykiya) log cfu/mL of fermented cow milk (mean ± standard error). The numbers of Bifidobacterium spp., Lb. plantarum, Lb. helveticus, and Lb. acidophilus revealed no significant difference between the 3 regions; nevertheless, Lactobacillus paracasei, Lactobacillus fermentum, Lactobacillus sakei, and Lactobacillus delbrueckii ssp. bulgaricus exhibited different degrees of variation across 3 regions. The results demonstrate that traditional fermented dairy products from different regions of Russia have complex compositions of LAB species. The diversity of LAB might be related to the type of fermented dairy product, geographical origin, and manufacturing process.

Diversity of lactic acid bacteria associated with traditional fermented dairy products in Mongolia.

Spontaneous milk fermentation has a long history in Mongolia, and beneficial microorganisms have been handed down from one generation to the next for use in fermented dairy products. The objective of this study was to investigate the diversity of lactic acid bacteria (LAB) communities in fermented yak, mare, goat, and cow milk products by analyzing 189 samples collected from 13 different regions in Mongolia. The LAB counts in these samples varied from 3.41 to 9.03 log cfu/mL. Fermented yak and mare milks had almost identical mean numbers of LAB, which were significantly higher than those in fermented goat milk but slightly lower than those in fermented cow milk. In total, 668 isolates were obtained from these samples using de Man, Rogosa, and Sharpe agar and M17 agar. Each isolate was considered to be presumptive LAB based on gram-positive and catalase-negative properties, and was identified at the species level by 16S rRNA gene sequencing, multiplex PCR assay, and restriction fragment length polymorphism analysis. All isolates from Mongolian dairy products were accurately identified as Enterococcus faecalis (1 strain), Enterococcus durans (3 strains), Lactobacillus brevis (3 strains), Lactobacillus buchneri (2 strains), Lactobacillus casei (16 strains), Lactobacillus delbrueckii ssp. bulgaricus (142 strains), Lactobacillus diolivorans (17 strains), Lactobacillus fermentum (42 strains), Lactobacillus helveticus (183 strains), Lactobacillus kefiri (6 strains), Lactobacillus plantarum ssp. plantarum (7 strains), Lactococcus lactis ssp. lactis (7 strains), Leuconostoc lactis (22 strains), Leuconostoc mesenteroides (21 strains), Streptococcus thermophilus (195 strains), and Weissella cibaria (1 strain). The predominant LAB were Strep. thermophilus and Lb. helveticus, which were isolated from all sampling sites. The results demonstrate that traditional fermented dairy products from different regions of Mongolia have complex compositions of LAB species. Such diversity of LAB provides useful information for further studies of probiotic strain selection and starter culture design, with regard to the industrial production of traditional fermented milk.