Cold growth behaviour and genetic comparison of Canadian and Swiss Listeria monocytogenes strains associated with the food supply chain and human listeriosis cases.

Sixty-two strains of Listeria monocytogenes isolated in Canada and Switzerland were investigated. Comparison based on molecular genotypes confirmed that strains in these two countries are genetically diverse. Interestingly strains from both countries displayed similar range of cold growth phenotypic profiles. Based on cold growth lag phase duration periods displayed in BHI at 4 °C, the strains were similarly divided into groups of fast, intermediate and slow cold adaptors. Overall Swiss strains had faster exponential cold growth rates compared to Canadian strains. However gene expression analysis revealed no significant differences between fast and slow cold adapting strains in the ability to induce nine cold adaptation genes (lmo0501, cspA, cspD, gbuA, lmo0688, pgpH, sigB, sigH and sigL) in response to cold stress exposure. Neither was the presence of Stress survival islet 1 (SSI-1) analysed by PCR associated with enhanced cold adaptation. Phylogeny based on the sigL gene subdivided strains from these two countries into two major and one minor cluster. Fast cold adaptors were more frequently in one of the major clusters (cluster A), whereas slow cold adaptors were mainly in the other (cluster B). Genetic differences between these two major clusters are associated with various amino acid substitutions in the predicted SigL proteins. Compared to the EGDe type strain and most slow cold adaptors, most fast cold adaptors exhibited five identical amino acid substitutions (M90L, S203A/S203T, S304N, S315N, and I383T) in their SigL proteins. We hypothesize that these amino acid changes might be associated with SigL protein structural and functional changes that may promote differences in cold growth behaviour between L. monocytogenes strains.

Complete Genome Sequence of Listeria monocytogenes LL195, a Serotype 4b Strain from the 1983-1987 Listeriosis Epidemic in Switzerland.

The complete genome sequence of Listeria monocytogenes LL195, a serotype 4b clinical strain isolated during the 1983-1987 listeriosis epidemic in Switzerland, is presented.

Examination of food chain-derived Listeria monocytogenes strains of different serotypes reveals considerable diversity in inlA genotypes, mutability, and adaptation to cold temperatures.

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.

Microcistina en plantas de tratamiento de agua para consume humano en un ambiente tropical: el Area Metropolitana de Costa Rica / Microcystin in plants that treat water for human consumption in a tropical environment: the Metropolitan Area of Costa Rica

We measured microcystin levels in water of the Metropolitan Area of Costa Rica by competitive inhibition ELISA and we quantified total coliforms, fecal coliforms, Escherichia coli (by a Most Probable Number method) and aerobic count. We wanted to identify any cyanotoxin correlation with these parameters, as a public health risk. We sampled in the rainy season of 2003 (April-October) and in the dry season of 2004 (February-March) (30 samples/season). We sampled pre-treated, semi-treated and treated water. Microcystin levels < 0.5 ppb were found in the rainy season (and > 0.5 ppb in the dry season). Dry season levels exceeded World Health Organization limits (1.0 ppb). Cyanotoxins occurred in the Tres Rios plant. We did not find a correlation between these microbiologic parameters of water quality and microcystin levels in water.

Se midió la presencia de microcistina en el Área Metropolitana de Costa Rica por la técnica de ELISA de inhibición competitiva y se cuantificó coliformes totales, coliformes termotolerantes, Escherichia coli (por medio de la técnica Numero Más Probable) y recuento total aerobio. Se realizaron dos etapas de muestreo, una durante la estación lluviosa del 2003 (abril-octubre) y otra durante la estación seca del 2004 (febrero-marzo), cada una con 30 muestras de agua. Se muestreó agua pretratada, semitratada y tratada. Se determinaron niveles de microcistina < 0.5 ppb durante la estación lluviosa del 2003, mientras que durante la estación seca, se detectaron concentraciones de microcistina > 0.5 ppb. Se informó la presencia de cianotoxinas en la Planta de Tratamiento de Tres Ríos. No se establece correlación entre los parámetros microbiológicos de calidad del agua y las concentraciones de microcistina en el agua. Los estudios deberían considerar la diversidad y toxicidad de cianobacterias en estas plantas, los efectos del tratamiento, y presencia de otros microorganismos y sustancias (dióxido de carbono, fósforo, Nitrógeno), sobre la presencia, estructura y efecto de estas toxinas.