Comparison of cell invasion, macrophage survival and inflammatory cytokines profiles between Salmonella enterica serovars Enteritidis and Dublin from Brazil.

AIMS: This study compared the capacity of strains of Salmonella enterica serovars Enteritidis and Dublin isolated in Brazil to invade epithelial cells, to be internalized by and survive within macrophages, and to stimulate cytokine release in vitro. METHODS AND RESULTS: Both serovars infected 75 and 73% Caco-2 (human) and MDBK (bovine) epithelial cells respectively. Salmonella Dublin and S. Enteritidis (i) were internalized at the respective rates of 79·6 and 65·0% (P ≤ 0·05) by U937 (human) macrophages, and 70·4 and 66·9% by HD11 (chicken) macrophages; and (ii) multiplied at the respective rates of 3·2- and 2·7-fold within U937 cells, and 1·9- and 1·1-fold (P ≤ 0·05) within HD11 cells respectively. Seventy per cent of 10 S. Dublin strains stimulated IL-8 production, while 70% of S. Enteritidis strains enhanced production of IL-1ß, IL-6, IL-8, IL-10, IL-12p70 and TNF in Caco-2 cells. CONCLUSIONS: Compared with S. Enteritidis, S. Dublin had stronger ability to survive within macrophages and induced weak cytokine production, which may explain the higher incidence of invasive diseases caused by S. Dublin in humans. SIGNIFICANCE AND IMPACT OF THE STUDY: This study compared S. enterica serovars Enteritidis and Dublin to provide comparative data about the profile of the two serovars in cells from humans, the common host and their respective natural animal hosts and vice versa in order to check the differences between these two phylogenetically closely related serovars that share antigenic properties but present different phenotypic behaviours.

Genetic diversity, anti-microbial resistance, plasmid profile and frequency of the Vi antigen in Salmonella Dublin strains isolated in Brazil.

Salmonella Dublin is strongly adapted to cattle causing enteritis and/or systemic disease with high rates of mortality. However, it can be sporadically isolated from humans, usually causing serious disease, especially in patients with underlying chronic diseases. The aim of this study was to molecularly type S. Dublin strains isolated from humans and animals in Brazil to verify the diversity of these strains as well as to ascertain possible differences between strains isolated from humans and animals. Moreover, the presence of the capsular antigen Vi and the plasmid profile was characterized in addition to the anti-microbial resistance against 15 drugs. For this reason, 113 S. Dublin strains isolated between 1983 and 2016 from humans (83) and animals (30) in Brazil were typed by PFGE and MLVA. The presence of the capsular antigen Vi was verified by PCR, and the phenotypic expression of the capsular antigen was determined serologically. Also, a plasmid analysis for each strain was carried out. The strains studied were divided into 35 different PFGE types and 89 MLVA-types with a similarity of ≥80% and ≥17.5%, respectively. The plasmid sizes found ranged from 2 to >150 kb and none of the strains studied presented the capsular antigen Vi. Resistance or intermediate resistance was found in 23 strains (20.3%) that were resistant to ampicillin, ciprofloxacin, chloramphenicol, imipenem, nalidixic acid, piperacillin, streptomycin and/or tetracycline. The majority of the S. Dublin strains studied and isolated over a 33-year period may descend from a common subtype that has been contaminating humans and animals in Brazil and able to cause invasive disease even in the absence of the capsular antigen. The higher diversity of resistance phenotypes in human isolates, as compared with animal strains, may be a reflection of the different anti-microbial treatments used to control S. Dublin infections in humans in Brazil.

Virulence-associated genes, antimicrobial resistance and molecular typing of Salmonella Typhimurium strains isolated from swine from 2000 to 2012 in Brazil.

AIMS: The aims of this study were to assess the pathogenic potential, antimicrobial resistance and genotypic diversity of Salmonella Typhimurium strains isolated in Brazil from swine (22) and the surrounding swine environment (5) from 2000 to 2012 and compare them to the profiles of 43 human strains isolated from 1983 to 2010, which had been previously studied. METHODS AND RESULTS: The presence of 12 SPI-1, SPI-2 and plasmid genes was assessed by PCR, the antimicrobial susceptibility to 13 antimicrobials was determined by the disc diffusion assay and genotyping was performed using pulsed-field gel electrophoresis (PFGE), multiple-locus variable-number of tandem repeats analysis (MLVA) and ERIC-PCR. More than 77·8% of the swine strains carried 10 or more of the virulence markers. Ten (37%) strains isolated from swine were multi-drug resistant (MDR). All the molecular typing techniques grouped the strains in two main clusters. Some strains isolated from swine and humans were allocated together in the PFGE-B2, MLVA-A1, MLVA-B and ERIC-A1 clusters. CONCLUSIONS: The genotyping results suggest that some strains isolated from swine and humans may descend from a common subtype and may indicate a possible risk of MDR S. Typhimurium with high frequency of virulence genes isolated from swine to contaminate humans in Brazil. SIGNIFICANCE AND IMPACT OF THE STUDY: This study provided new information about the pathogenic potential, antimicrobial resistance and genotypic diversity of S. Typhimurium isolates from swine origin in Brazil, the fourth largest producer of pigs worldwide.

Genotypic diversity and pathogenic potential of Yersinia enterocolitica biotype 2 strains isolated in Brazil.

AIMS: To investigate the pathogenic potential and genotypic diversity of Yersinia enterocolitica biotype 2 strains isolated in Brazil and to compare these strains with other Y. enterocolitica biotypes using ERIC-PCR and PFGE. METHODS AND RESULTS: Forty strains of Y. enterocolitica biotype 2 (B2) isolated from humans (5), the environment (34) and animal (1), in Brazil over 19 years were studied. In addition to these isolates, we also analysed 26 Y. enterocolitica strains belonging to the biotypes 1A, 1B, and 3-5. All of the B2 strains contained the genes inv, ail, ystA, hreP, tccC and myfA. The genes fepD and fes were detected in 39 (97·5%) strains, virF was found in three (7·5%) strains, and ystB and fepA were not detected in any strains. The B2 strains showed genotypic similarities of more than 84·8% by ERIC-PCR and of more than 69·0% by PFGE. CONCLUSIONS: The pathogenic potential of the B2 strains examined in this study was highlighted by the occurrence of the majority of the virulence markers searched. The results of the ERIC-PCR and PFGE showed that the B2 strains evaluated in this study had a high genotypic similarity, suggesting that these strains differed little over the 19 year study period and that the environment was a possible source of contamination of humans and animals in Brazil. Furthermore, the ERIC-PCR technique grouped the strains belonging to Y. enterocolitica biotypes 1A, 1B, 2, 3, 4 and 5 according to their pathogenicity. SIGNIFICANCE AND IMPACT OF THE STUDY: This study provided new information about the pathogenic potential and genotypic similarity of Y. enterocolitica B2 isolated from diverse sources in Brazil. Furthermore, ERIC-PCR showed to be a valuable tool for grouping Y. enterocolitica of different biotypes according their pathogenicity.

Characterization of Salmonella Enteritidis strains isolated from poultry and farm environments in Brazil.

Salmonella Enteritidis is a major causative agent of foodborne outbreaks worldwide. Using enterobacterial repetitive intergenic consensus-polymerase chain reaction (ERIC-PCR) and pulsed-field gel electrophoresis (PFGE), this study assessed the genetic relatedness, the pathogenic potential, and antimicrobial resistance in 60 strains isolated from chickens and the farm environment in Brazil between 2004 and 2010. The resulting concatenated dendrogram of the two methodologies distinguished the strains into two clusters. Some strains isolated from the two sources were indistinguishable. All the strains contained the 13 virulence markers investigated. Forty-four strains were resistant to nalidixic acid. Quinolone resistance presented by many strains suggests that quinolones may have been used to treat chickens. The high prevalence of virulence markers highlights the importance of poultry as vehicles of S. Enteritidis strains that have the potential to cause disease.

Molecular typing of Yersinia pseudotuberculosis strains isolated from livestock in Brazil.

Yersinia pseudotuberculosis can infect a broad range of animals. In Brazil, this bacterium has been isolated from healthy and sick animals from sporadic cases and outbreaks of hemorrhagic gastroenteritis among livestock. However, the molecular diversity of these isolates is little understood. In this study, we used multilocus sequence typing, enterobacterial repetitive intergenic consensus PCR and pulsed-field gel electrophoresis to genotype 40 Y. pseudotuberculosis strains belonging to bio-serogroups 1/O:1a and 2/O:3 isolated between 1982 and 1990 in the southern region of Brazil. All three methodologies clustered the strains into two main clusters according to their bio-serogroups. Good correlations were observed between the clusters and the pathogenic potential of the strains. No correlation among the strains was observed according to geographical origin, host, place, or year of isolation. The grouping of the Y. pseudotuberculosis isolated in Brazil determined by these assays leads us to suggest that Brazilian livestock harbor two subpopulations of Y. pseudotuberculosis.

Panoramic view of the occurrence of Yersinia species other than Y. pestis in Brazil

Data on the occurrence of Yersinia species. other than Y. pestis in Brazil are presented. Over the past 40 years, 767 Yersinia strains have been identified and typed by the National Reference Center on Yersinia spp. other than Y. pestis, using the classical biochemical tests for species characterization. The strains were further classified into biotypes, serotypes and phagetypes when pertinent. These tests led to the identification of Yersinia cultures belonging to the species Y. enterocolitica, Y.pseudotuberculosis, Y. intermedia, Y. frederiksenii and Y. kristensenii. Six isolates could not be classified in any of the known Yersinia species and for this reason were defined as Non-typable (NT). The bio-sero-phagetypes of these strains were diverse. The following species of Yersinia were not identified among the Brazilian strains by the classical phenotypic or biochemical tests: Y. aldovae, Y. rhodei, Y. mollaretti, Y. bercovieri and Y.ruckeri. The Yersinia strains were isolated from clinical material taken from sick and/or healthy humans and animals, from various types of food and from the environment, by investigators of various Institutions localized in different cities and regions of Brazil.

Plesiomonas shigelloides: um enteropatógeno emergente? / Plesiomonas shigelloides: an emergent enteropathogen?

Plesiomonas shigelloides é um bacilo Gram-negativo, pertencente à família Enterobacteriaceae, isolado de água doce e salgada, de peixes de água doce, mariscos e de inúmeros tipos de animais. Suspeita-se que a maioria das infecções humanas causadas por P. shigelloides, seja veiculada pela água, pois a bactéria está presente em águas não tratadas que são usadas para beber, águas recreacionais ou água para lavar alimentos que são consumidos sem cozimento ou aquecimento. A ingestão de P. shigelloides não causa sempre doença no animal hospedeiro, mas o microrganismo pode permanecer temporariamente como membro transitório não infeccioso da microbiota intestinal. A bactéria é isoladade fezes de pacientes com diarréia, mas algumas vezes também de fezes de indivíduos sem sintomas. A doença causada por P. shigelloides é a gastrenterite, que normalmente é auto-limitante, com febre, calafrio, dor abdominal, náusea, diarréia ou vômito. Em casos graves, as fezes diarréicas podem ser verde-amareladas, espumosas e com presença de sangue. A bactéria pode também causar infecções extra-intestinais. Ademais, pode produzir toxinas e ser invasora. As características utilizadas para considerar P. shigelloides como um enteropatógeno não são totalmente convincentes. Embora seja isolada de pacientes com diarréia e incriminada em vários surtos epidêmicos envolvendo água e alimentos contaminados, não foi possível identificar em muitas amostras de P. shigelloides, associadas com infecções gastrintestinais, um mecanismo de virulência definitivo.

Importância de Yersinia enterocolitica em microbilogia médica / Importance of Yersinia enterocolitica in medical microbiology

Y.enterocolitica é um enteropatógeno invasivo de humanos que provoca uma série de sintomas clínicos intestinais e extra-intestinais que variam desde uma gastrenterite branda a uma linfadenite mesentérica que mimetiza apendicite e em casos raros pode evoluir para uma septicemia.A infecção causada por Y.enterocolitica pode levar a seqüelas imunológicas, incluindo artrite, eritema nodoso e glomerulonefrite.Amostras patogênicas de Y.enterocolitica são associadas a determinados sorogrupos e biotipos e a uma variedade de características fenotípicas relacionadas a virulência.Estudos de genética molecular demonstraram a importância do plasmídio pYV que codifica vários genes de virulência, bem como a importância de vários genes de virulência cromossomais na patogênese dessa bactéria.As infecções intestinais causadas por Y.enterocolitica são normalmente auto-limitadas não havendo usualmente a necessidade de antibioticoterapia.A ocorrência de infecções por Y.enterocolitica no Brasil não é tão freqüente como em países europeus, Japão e Estados Unidos.Essa revisão enfoca as características gerais, a patogênese, os sintomas clínicos, mecanismos de virulência, tratamento e susceptibilidade a antibióticos de amostras de Y.enterocolitica isoladas no Brasil e ao redor do mundo.

Virulence characteristics and epidemiology of Yersinia enterocolitica and Yersiniae other than Y. pseudotuberculosis and Y. pestis isolated from water and sewage.

AIMS: To determine the species, bio-sero-phagetypes, antimicrobial drug resistance and also the pathogenic potential of 144 strains of Yersinia spp. isolated from water sources and sewage in Brazil. METHODS AND RESULTS: The 144 Yersinia strains were characterized biochemically, serologically and had their antibiotic resistance and phenotypic virulence markers determined by microbiological and serological standard techniques. The Y. enterocolitica strains related to human diseases were also tested for the presence of virulence genes, by the PCR technique. The isolates were classified as Y. enterocolitica, Y. intermedia, Y. frederiksenii, Y. kristensenii and Yersinia biochemically atypical. The 144 isolates belonged to various bio-serogroups. Half of the strains showed resistance to three or more drugs. The Y. enterocolitica strains related to human diseases exhibited phenotypic virulence characteristics and virulence genes. CONCLUSIONS: Water from various sources and sewage are contaminated with Yersinia spp. in Brasil. Among these bacteria, virulent strains of Y. enterocolitica were found, with biotypes and serogroups related to human diseases. SIGNIFICANCE AND IMPACT OF THE STUDY: This is the first documented description of the occurrence of pathogenic Y. enterocolitica in water sources and sewage in Brazil. The occurrence of virulence strains of Y. enterocolitica shows that the environment is a potential source of human infection by this species in this country.