Centro Nacional Patagónico, CENPAT-CONICET, Puerto Madryn, Chubut, Argentina.
Arch Microbiol. 2010 Apr;192(4):237-45. doi: 10.1007/s00203-010-0552-1. Epub 2010 Feb 6.
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.
富集奥多尼斯肠内容物后,分离出 53 株乳酸菌(LAB)。从对乳球菌 03/8460 表现出抑制活性的 4 个分离株中,选择了 TW34 菌株,因为它的抑制作用最强。它还抑制其他革兰氏阳性菌,但不抑制革兰氏阴性鱼类病原体。表型和 16S rDNA 系统发育分析表明,TW34 属于乳球菌 lactis。此外,TW34 对不同的抗生素表现出敏感性。对 L. garvieae 的抑制因子的产生与生长相关,并且受孵育温度的显著影响。抗菌生产的最佳温度低至 15 摄氏度。酸化和过氧化氢的产生都不是抑制的来源。相比之下,用蛋白酶处理完全丧失了抗菌活性,这证实了抑制物质是一种细菌素。细菌素具有高度的热稳定性(121°C 下 15 分钟),在 pH 3 到 11 之间具有活性。在 4°C 下储存长达 2 个月,在-20°C 下储存长达 6 个月时保持稳定。我们的结果表明,L. lactis TW34 菌株可提供控制乳球菌病的替代方法,因此可考虑用于未来的鱼类挑战实验。