Medical & Bio-Materials Research Center and Division of Biomaterials Engineering, Kangwon National University, Chucheon, Gangwon 200-701, Republic of Korea.
Appl Environ Microbiol. 2010 Dec;76(24):7910-7. doi: 10.1128/AEM.01508-10. Epub 2010 Oct 22.
This study was designed to characterize the viability and potential virulence of bofilm-forming Salmonella enterica serovar Typhimurium under different pH levels, ranging from 5 to 7. The plate count method and real-time reverse transcription-PCR (RT-PCR) were used to evaluate the survival of S. Typhimurium grown in Trypticase soy broth (TSB) adjusted to pH 5, 6, and 7 (TSB-5, TSB-6, and TSB-7, respectively) at 37°C for 10 days. In TSB-5 and TSB-6, the numbers of viable cells estimated by using the real-time RT-PCR were greater than the culturable counts enumerated by the plate count method. Reflectance micro-Fourier transform infrared (micro-FTIR) spectroscopy was used to evaluate the biochemical changes in biofilm cells. Considerable changes in chemical components were observed in the biofilm cells grown in TSB-5 and TSB-6 when compared to the cells grown in TSB-7. The enterotoxin production and invasive ability of planktonic and biofilm S. Typhimurium cells were inferred by the relative levels of expression of stn and invA. The levels of expression of stn and invA were significantly increased in biofilm S. Typhimurium cells grown in TSB-5 (1.9-fold and 3.2-fold) and TSB-6 (2.1-fold and 22.3-fold) after 10 days of incubation. These results suggest that the biofilm-forming S. Typhimurium under different pH levels might change the virulence production and stress response mechanisms.
本研究旨在表征不同 pH 值(5 至 7)条件下形成生物膜的鼠伤寒沙门氏菌血清型 Typhimurium 的生存能力和潜在毒力。采用平板计数法和实时逆转录聚合酶链反应(RT-PCR)评估了在 37°C 下于 pH 值分别为 5、6 和 7 的胰蛋白胨大豆肉汤(TSB)(分别为 TSB-5、TSB-6 和 TSB-7)中生长 10 天的鼠伤寒沙门氏菌的存活情况。在 TSB-5 和 TSB-6 中,使用实时 RT-PCR 估计的活菌数大于平板计数法枚举的可培养数。反射微傅里叶变换红外(micro-FTIR)光谱用于评估生物膜细胞的生化变化。与在 TSB-7 中生长的细胞相比,在 TSB-5 和 TSB-6 中生长的生物膜细胞的化学成分发生了相当大的变化。通过 stn 和 invA 的相对表达水平推断出浮游和生物膜鼠伤寒沙门氏菌细胞的肠毒素产生和侵袭能力。在 TSB-5(1.9 倍和 3.2 倍)和 TSB-6(2.1 倍和 22.3 倍)中培养 10 天后,生物膜鼠伤寒沙门氏菌细胞中 stn 和 invA 的表达水平显著增加。这些结果表明,不同 pH 值下形成生物膜的鼠伤寒沙门氏菌可能会改变其毒力产生和应激反应机制。