806

Microbial analysis of ovine cheese by next generation sequencing

Vladimir Kmet ✉, Dobroslava Bujnakova

Institute of Animal Physiology, Slovak Academy of Sciences, Košice, Slovakia

Kmet V and Bujnakova D (2015) EMBnet.journal 21(Suppl A), e806. http://dx.doi.org/10.14806/ej.21.A.806

Ovine cheese is a good source for isolation of wild lactobacilli-potential starter cultures. This study aimed to analyse microbiota of the ovine cheese (Slovak Bryndza) and to investigate the presence of Lactobacillus antibiotic resistance, virulence or probiotic genes by pyrosequencing.

The V1-V3 regions of the 16S ribosomal DNA were amplifed from different ovine cheeses using PCR. In all samples, the microbial populations consisted of Lactobacillus helveticus, Lb. acidophilus, Lb. plantarum, Lb. rhamnosus, Lb. brevis; Lactococcus lactis, L. raffinolactis, L. garviae; Enterococcus italicus and E. cameliae; Streptococcus salivarius, St. thermophilus, St. caballi and St. ferus.

Furthermore, the genomes of selected Lb. plantarum, Lb. brevis, Lb. paracasei were pyrosequenced. The assembly of L. plantarum resulted in 203 contigs longer than 1,000 bp (D’Auria et al., 2014). There were identified probiotic proteins as an alpha amylase (PF00128), peptidase (PF01433), catalase (PF00199), heat shock protein 33 (PF01430). Nevertheless, there was discrepancy between Lb. plantarum ampicillin sensitivity and the presence of serine beta-lactamase like superfamily (PF00144). No virulence factors were detected. Results indicated new properties of lactobacilli, which were not occurred by phenotyping testing.

Acknowledgements

This work was supported by Slovak grant VEGA No. 2/0014/13.

References

D’Auria G, Džunkova M, Moya A, Tomaška M, Kološta M, Kmet V (2014). Genome Sequence of Lactobacillus plantarum 19L3, a Strain Proposed as a Starter Culture for Slovenska Bryndza Ovine Cheese. Genome Announc. 2(2):e00292-14. http://dx.doi.org/10.1128/genomeA.00292-14

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