Food, Nutrition and Health Program, Faculty of Land and Food Systems, University of British Columbia, Vancouver, BC, Canada.
Appl Environ Microbiol. 2013 Mar;79(6):1915-22. doi: 10.1128/AEM.03341-12. Epub 2013 Jan 11.
Listeria monocytogenes strains belonging to serotypes 1/2a and 4b are frequently linked to listeriosis. While inlA mutations leading to premature stop codons (PMSCs) and attenuated virulence are common in 1/2a, they are rare in serotype 4b. We observed PMSCs in 35% of L. monocytogenes isolates (n = 54) recovered from the British Columbia food supply, including serotypes 1/2a (30%), 1/2c (100%), and 3a (100%), and a 3-codon deletion (amino acid positions 738 to 740) seen in 57% of 4b isolates from fish-processing facilities. Caco-2 invasion assays showed that two isolates with the deletion were significantly more invasive than EGD-SmR (P < 0.0001) and were either as (FF19-1) or more (FE13-1) invasive than a clinical control strain (08-5578) (P = 0.006). To examine whether serotype 1/2a was more likely to acquire mutations than other serotypes, strains were plated on agar with rifampin, revealing 4b isolates to be significantly more mutable than 1/2a, 1/2c, and 3a serotypes (P = 0.0002). We also examined the ability of 33 strains to adapt to cold temperature following a downshift from 37°C to 4°C. Overall, three distinct cold-adapting groups (CAG) were observed: 46% were fast (<70 h), 39% were intermediate (70 to 200 h), and 15% were slow (>200 h) adaptors. Intermediate CAG strains (70%) more frequently possessed inlA PMSCs than did fast (20%) and slow (10%) CAGs; in contrast, 87% of fast adaptors lacked inlA PMSCs. In conclusion, we report food chain-derived 1/2a and 4b serotypes with a 3-codon deletion possessing invasive behavior and the novel association of inlA genotypes encoding a full-length InlA with fast cold-adaptation phenotypes.
李斯特菌单核细胞增生李斯特菌血清型 1/2a 和 4b 菌株常与李斯特菌病有关。虽然 1/2a 中常见导致提前终止密码子(PMSCs)和毒力减弱的 inlA 突变,但在血清型 4b 中很少见。我们观察到不列颠哥伦比亚省食品供应中回收的 54 株李斯特菌分离株中有 35%(n=54)存在 PMSCs,包括血清型 1/2a(30%)、1/2c(100%)和 3a(100%),以及在鱼类加工设施中 4b 分离株中 57%存在的 3 个密码子缺失(氨基酸位置 738 至 740)。Caco-2 侵袭实验表明,两个缺失的分离株的侵袭性明显高于 EGD-SmR(P<0.0001),与临床对照株(08-5578)相比,侵袭性分别为(FF19-1)或更高(FE13-1)(P=0.006)。为了研究血清型 1/2a 是否比其他血清型更易发生突变,我们将菌株接种在含有利福平的琼脂上,结果发现 4b 分离株比 1/2a、1/2c 和 3a 血清型更容易发生突变(P=0.0002)。我们还研究了 33 株菌株在从 37°C 下降到 4°C 后适应低温的能力。总的来说,观察到三个不同的低温适应群(CAG):46%是快速(<70 h),39%是中间(70 至 200 h),15%是缓慢(>200 h)适应者。中间 CAG 菌株(70%)比快速(20%)和缓慢(10%)CAG 菌株更频繁地携带 inlA PMSCs;相比之下,87%的快速适应者缺乏 inlA PMSCs。总之,我们报告了食物链衍生的 1/2a 和 4b 血清型具有 3 个密码子缺失的侵袭性行为,以及 inlA 基因型编码全长 InlA 与快速低温适应表型的新关联。