Inhibitory activity against the fish pathogen Lactococcus garvieae produced by Lactococcus lactis TW34, a lactic acid bacterium isolated from the intestinal tract of a Patagonian fish.

After enrichment of Odontesthes platensis intestinal contents, 53 lactic acid bacteria (LAB) were isolated. From the four isolates that showed inhibitory activity against Lactococcus garvieae 03/8460, strain TW34 was selected because it exerted the strongest inhibition. It also inhibited other Gram-positive bacteria, but not Gram-negative fish pathogens. Phenotypic and 16S rDNA phylogenetic analyses showed that TW34 belongs to Lactococcus lactis. In addition, TW34 showed to be sensitive to different antibiotics. The production of the inhibitory agent against L. garvieae was growth associated, and it was significantly influenced by the incubation temperature. The optimal temperature for the antimicrobial production was as low as 15 degrees C. Both acidification and hydrogen peroxide production were ruled out as the source of inhibition. In contrast, the antimicrobial activity was completely lost by treatment with proteolytic enzymes, which confirmed that the inhibitory substance was a bacteriocin. The bacteriocin was highly thermostable (121 degrees C for 15 min) and active between pH 3 and 11. It remained stable for up to 2 months when stored at 4 degrees C and up to 6 months at -20 degrees C. Our results suggest that the strain L. lactis TW34 could provide an alternative for lactococcosis control and therefore be considered for future challenge experiments with fish.

Factores de virulencia de cepas de Enterococcus aisladas de quesos ovinos / Virulence factors of Enterococcus strains isolated from ovine cheese

Los enterococos son utilizados en la industria alimenticia como cultivos iniciadores o probióticos y constituyen contaminantes naturales de los alimentos. Sin embargo, el género Enterococcus ha cobrado relevancia como causal de infecciones nosocomiales, tendencia exacerbada por el desarrollo de resistencia antibiótica. Con el objetivo de estudiar la virulencia potencial de ocho cepas de Enterococcus faecium y de dos cepas de Enterococcus faecalis aislados de quesos ovinos se investigó la resistencia a vancomicina, la actividad hemolítica y la actividad de gelatinasa. En forma adicional se llevó a cabo la reacción en cadena de la polimerasa (PCR) para determinar la presencia de los genes cylB de la citolisina, gelE de la gelatinasa, cpd de la feromona sexual y agg de la proteína de agregación. Ninguna de las cepas mostró resistencia a la vancomicina o actividad hemolítica. El gen cylB no pudo ser identificado mediante amplificación por PCR en ninguna de las cepas estudiadas. La presencia del gen gelE fue detectada en siete cepas de E. faecium y en una cepa de E. faecalis, sin embargo en ningún caso se detectó actividad de la enzima. El gen cpd fue detectado en E. faecium ETw7 y E. faecalis ETw23, mientras que el gen agg fue hallado en las cepas de E. faecium ETw7 y E. faecalis ETw27. Estos resultados sugieren que la introducción de productos alimenticios o probióticos basados en el uso de cepas de enterococos requiere una cuidadosa evaluación sobre su seguridad.

Enterococci are used as starter and probiotic cultures in the food industry, and they occur as natural food contaminants. However, the genus Enterococcus is of increased significance as a cause of nosocomial infections, exacerbated by the development of antibiotic resistance. In order to study the potential virulence of eight Enterococcus faecium strains and two Enterococcus faecalis strains isolated from ovine cheese, vancomicine resistance, hemolytic activity and gelatinase activity were investigated. In addition, polymerase chain reaction (PCR) tests were carried out in order to determine the presence of cytolicyn gene cylB, gelatinase gene gelE, sex pheromone gene cpd and aggregation protein gene agg. None of the strains showed vancomicine resistance or hemolitic activity. Gene cylB could not be identified by PCR amplification in any of the strains studied. The presence of gene gelE was found in seven E. faecium strains and in one E. faecalis strain, however in no case was gelatinase activity detected. Gene cpd was detected in E. faecium ETw7 and E. faecalis ETw23, while gene agg was found in E. faecium ETw7 and E. faecalis ETw27. These results suggest that the introduction of food products or probiotics based on the use of enterococal strains requires careful safety evaluations.