Andrews Robert E, Johnson Wesley S, Guard Abby R, Marvin Jonathan D
Department of Ecology, Evolution and Organismal Biology, Iowa State University, Ames, IA 50011, USA.
Can J Microbiol. 2004 Nov;50(11):957-66. doi: 10.1139/w04-090.
The persistence of Enterococcus faecalis, fecal enterococci from swine waste, and Tn916-like elements was determined following inoculation into autoclaved and native soil microcosms. When cells of E. faecalis CG110 (Tn916) were inoculated into native microcosms, enterococcal viability in the soil decreased approximately 5 orders of magnitude (4.8 x 10(5) CFU/g soil to < 10 CFU/g) after 5 weeks. In autoclaved microcosms, the viability of E. faecalis decreased by only 20% in 5 weeks. In contrast, the content of Tn916, based on PCR of DNA extracts from soil microcosms, decreased by about 20% in both native and autoclaved microcosms. Similar results were obtained when the source of fecal enterococci and Tn916-like elements was swine waste. Because the concentration of Tn916-independent E. faecalis DNA (the D-alanine D-alanine ligase gene), based on PCR, decreased to nearly undetectable levels (at least 3 orders of magnitude) after 5 weeks in the native microcosms, the evidence suggests Tn916 stability in the soil results from en masse transfer of the transposon to the normal soil microflora and not survival of E. faecalis DNA in the soil system. Results from denaturing gradient gel electrophoresis suggest that multiple forms of Tn916 occur in swine waste, but only forms most like Tn916 exhibit stability in the soil.
将粪肠球菌、猪粪便中的粪肠球菌和类Tn916元件接种到高压灭菌的天然土壤微观环境中后,测定了它们的持久性。当将粪肠球菌CG110(Tn916)的细胞接种到天然微观环境中时,5周后土壤中肠球菌的活力下降了约5个数量级(从4.8×10⁵CFU/g土壤降至<10CFU/g)。在高压灭菌的微观环境中,粪肠球菌的活力在5周内仅下降了20%。相比之下,基于土壤微观环境DNA提取物的PCR检测,Tn916的含量在天然和高压灭菌的微观环境中均下降了约20%。当粪肠球菌和类Tn916元件的来源为猪粪便时,也得到了类似的结果。由于基于PCR检测,在天然微观环境中5周后,不依赖Tn916的粪肠球菌DNA(D-丙氨酸-D-丙氨酸连接酶基因)浓度降至几乎无法检测的水平(至少3个数量级),因此有证据表明,土壤中Tn916的稳定性源于转座子整体转移至正常土壤微生物群落,而非粪肠球菌DNA在土壤系统中的存活。变性梯度凝胶电泳结果表明,猪粪便中存在多种形式的Tn916,但只有最类似Tn916的形式在土壤中表现出稳定性。