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Blum S.E.,Kimron Veterinary Institute | Sela S.,Institute for Postharvest and Food science | Elad D.,Kimron Veterinary Institute | Edery N.,Kimron Veterinary Institute | Leitner G.,Kimron Veterinary Institute
PLoS ONE | Year: 2015

Escherichia coli is a major etiological agent of intra-mammary infections (IMI) in cows, leading to acute mastitis and causing great economic losses in dairy production worldwide. Particular strains cause persistent IMI, leading to recurrent mastitis. Virulence factors of mammary pathogenic E. coli (MPEC) involved pathogenesis of mastitis as well as those differentiating strains causing acute or persistent mastitis are largely unknown. This study aimed to identify virulence markers in MPEC through whole genome and phenome comparative analysis. MPEC strains causing acute (VL2874 and P4) or persistent (VL2732) mastitis were compared to an environmental strain (K71) and to the genomes of strains representing different E. coli pathotypes. Intra-mammary challenge in mice confirmed experimentally that the strains studied here have different pathogenic potential, and that the environmental strain K71 is non-pathogenic in the mammary gland. Analysis of whole genome sequences and predicted proteomes revealed high similarity among MPEC, whereas MPEC significantly differed from the non-mammary pathogenic strain K71, and from E. coli genomes from other pathotypes. Functional features identified in MPEC genomes and lacking in the non-mammary pathogenic strain were associated with synthesis of lipopolysaccharide and other membrane antigens, ferric-dicitrate iron acquisition and sugars metabolism. Features associated with cytotoxicity or intra-cellular survival were found specifically in the genomes of strains from severe and acute (VL2874) or persistent (VL2732) mastitis, respectively. MPEC genomes were relatively similar to strain K-12, which was subsequently shown here to be possibly pathogenic in the mammary gland. Phenome analysis showed that the persistent MPEC was the most versatile in terms of nutrients metabolized and acute MPEC the least. Among phenotypes unique to MPEC compared to the non-mammary pathogenic strain were uric acid and D-serine metabolism. This study reveals virulence factors and phenotypic characteristics of MPEC that may play a role in pathogenesis of E. coli mastitis. Copyright: © 2015 Blum et al.

Kroupitski Y.,Institute for Postharvest and Food science | Kroupitski Y.,Hebrew University | Brandl M.T.,Produce Safety and Microbiology Research Unit | Pinto R.,Institute for Postharvest and Food science | And 4 more authors.
Phytopathology | Year: 2013

Recurrent outbreaks of enteric illness linked to lettuce and a lack of efficacious strategies to decontaminate produce underscores the need for a better understanding of the molecular interactions of foodborne pathogens with plants. This study aimed at identifying Salmonella enterica genes involved in the persistence of this organism on postharvest lettuce during cold storage using recombinase-based in vivo expression technology (RIVET). In total, 37 potentially induced loci were identified in four distinct screenings. Knockout mutations in eight upregulated genes revealed that four of them have a role in persistence of the pathogen in this system. These genes included stfC, bcsA, misL, and yidR, encoding a fimbrial outer membrane usher, a cellulose synthase catalytic subunit, an adhesin of the autotransporter family expressed from the Salmonella pathogenicity island-3, and a putative ATP/GTP-binding protein, respectively. bcsA, misL, and yidR but not stfC mutants were impaired also in attachment and biofilm formation, suggesting that these functions are required for survival of S. enterica on post-harvest lettuce. This is the first report that MisL, which has a role in Salmonella binding to fibronectin in animal hosts, is involved also in adhesion to plant tissue. Hence, our study uncovered a new plant attachment factor in Salmonella and demonstrates that RIVET is an effective approach for investigating human pathogen-plant interactions in a post-harvest leafy vegetable. © 2013 The American Phytopathological Society.

Goudeau D.M.,U.S. Department of Agriculture | Parker C.T.,U.S. Department of Agriculture | Zhou Y.,U.S. Department of Agriculture | Sela S.,Institute for Postharvest and Food science | And 2 more authors.
Applied and Environmental Microbiology | Year: 2013

Fresh vegetables have been recurrently associated with salmonellosis outbreaks, and Salmonella contamination of retail produce has been correlated positively with the presence of soft rot disease. We observed that population sizes of Salmonella enterica serovar Typhimurium SL1344 increased 56-fold when inoculated alone onto cilantro leaves, versus 2,884-fold when coinoculated with Dickeya dadantii, a prevalent pathogen that macerates plant tissue. A similar trend in S. enterica populations was observed for soft-rotted lettuce leaves. Transcriptome analysis of S. enterica cells that colonized D. dadantii-infected lettuce and cilantro leaves revealed a clear shift toward anaerobic metabolism and catabolism of substrates that are available due to the degradation of plant cells by the pectinolytic pathogen. Twenty-nine percent of the genes that were upregulated in cilantro macerates were also previously observed to have increased expression levels in the chicken intestine. Furthermore, multiple genes induced in soft rot lesions are also involved in the colonization of mouse, pig, and bovine models of host infection. Among those genes, the operons for ethanolamine and propanediol utilization as well as for the synthesis of cobalamin, a cofactor in these pathways, were the most highly upregulated genes in lettuce and cilantro lesions. In S. Typhimurium strain LT2, population sizes of mutants deficient in propanediol utilization or cobalamin synthesis were 10- and 3-fold lower, respectively, than those of the wildtype strain in macerated cilantro (P<0.0002); in strain SL1344, such mutants behaved similarly to the parental strain. Anaerobic conditions and the utilization of nutrients in macerated plant tissue that are also present in the animal intestine indicate a niche overlap that may explain the high level of adaptation of S. enterica to soft rot lesions, a common postharvest plant disease. © 2013, American Society for Microbiology.

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