Proteomic Characterization of Antibiotic Resistance, and Production of Antimicrobial and Virulence Factors in Streptococcus Species Associated with Bovine Mastitis. Could Enzybiotics Represent Novel Therapeutic Agents Against These Pathogens?

Streptococcus spp. are major mastitis pathogens present in dairy products, which produce a variety of virulence factors that are involved in streptococcal pathogenicity. These include neuraminidase, pyrogenic exotoxin, and M protein, and in addition they might produce bacteriocins and antibiotic-resistance proteins. Unjustifiable misuse of antimicrobials has led to an increase in antibiotic-resistant bacteria present in foodstuffs. Identification of the mastitis-causing bacterial strain, as well as determining its antibiotic resistance and sensitivity is crucial for effective therapy. The present work focused on the LC–ESI–MS/MS (liquid chromatography–electrospray ionization tandem mass spectrometry) analysis of tryptic digestion peptides from mastitis-causing Streptococcus spp. isolated from milk. A total of 2706 non-redundant peptides belonging to 2510 proteins was identified and analyzed. Among them, 168 peptides were determined, representing proteins that act as virulence factors, toxins, anti-toxins, provide resistance to antibiotics that are associated with the production of lantibiotic-related compounds, or play a role in the resistance to toxic substances. Protein comparisons with the NCBI database allowed the identification of 134 peptides as specific to Streptococcus spp., while two peptides (EATGNQNISPNLTISNAQLNLEDKNK and DLWC*NM*IIAAK) were found to be species-specific to Streptococcus dysgalactiae. This proteomic repository might be useful for further studies and research work, as well as for the development of new therapeutics for the mastitis-causing Streptococcus strainsThis work was supported by the Xunta de Galicia; the European Union Social Fund (ESF); the Spanish Ministry of Economy and Competitiveness (Project AGL 2.013-48.244-R); and by the European Regional Development Fund (ERDF) (2007-2013)S

Caracterización molecular de bacterias ácido-lácticas aisladas a partir de carne de ternera envasada al vacío de modo tradicional y mediante un sistema avanzado

A total of 91 lactic acid bacteria (LAB) were isolated from 50 beef samples, 25 packaged by traditional vacuum packaging and 25 packaged using advanced vacuum skin packaging. The isolated LAB were identified using 16S rRNA sequencing, whereas randomly amplified polymorphic DNA (RAPD-PCR) and cluster analysis were used for typing the LABs. Ten different species of LAB were identified and assigned to the following species: Enterococcus gilvus (22 isolates), Enterococcus faecium (9 isolates), Enterococcus casseliflavus (8 isolates), Enterococcus faecalis (4 isolates), Enterococcus malodoratus (3 isolates), Enterococcus devriessei (3 isolates), Lactobacillus sakei (15 isolates), Carnobacterium divergens (12 isolates), Carnobacterium maltaromaticum (5 isolates), and Leuconostoc mesenteroides (8 isolates). The RAPD profile bands observed were not significantly discriminatory, with exceptions of E. casseliflavus, E. faecalis, and E. faecium, which suggests that the type of packaging system used had no specific effect on the selection of most microbiota in the meat after packagingUn total de 91 bacterias ácido-lácticas (LAB) fueron aisladas a partir de 50 muestras de carne de ternera, de las cuales 25 fueron envasadas al vacío de modo tradicional y 25 fueron envasadas mediante un sistema de vacío avanzado. Los aislamientos fueron identificados a través de la secuenciación del ADN ribosomal 16S, mientras que la caracterización de las LAB se realizó mediante la amplificación al azar del ADN polimórfico (RAPD-PCR) y su posterior análisis cluster. Fueron identificadas cepas pertenecientes a diez especies bacterianas diferentes, en concreto: Enterococcus gilvus (22 aislamientos), E. faecium (9 aislamientos), E. casseliflavus (8 aislamientos), E. faecalis (4 aislamientos), E. malodoratus (3 aislamientos), E. devriessei (3 aislamientos), Lactobacillus sakei (15 aislamientos), Carnobacterium divergens (12 aislamientos), C. maltaromaticum (5 aislamientos) y Leuconostoc mesenteroides (8 aislamientos). Los perfiles de bandas de ADN obtenidas no revelaron diferencias significativas, con las excepciones de E. casseliflavus, E. faecalis y E. faecium, sugiriendo que el tipo de envasado no tiene un efecto específico en la selección de la mayor parte de la microbiota presente en la carne envasadaS