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.

Invasin of Yersinia pseudotuberculosis is not a polyclonal activator of mouse B lymphocytes

It is known that the invasin molecule of Yersinia pseudotuberculosis stimulates human peripheral B cells in vitro. In this work we evaluated the in vivo role ofinvasin as polyclonal activator of B lymphocytes in the mouse experimental model, by comparing strains of Y.pseudotuberculosis expressing invasin and isogenic inv mutants. Swiss mice were infected intravenously with two strains expressing invasin (YpIII pIB1 and an isogenic virulence plasmid-cured strain, YpIII) and with two invasin mutant strains (Yp100 pIB1 and Yp100, plasmid-cured). Spleen cells were sampled on days 7, 14, 21 and 28 after infection. Immunoglobulin (Ig)-secreting spleen cells were detected by protein A plaque assay and specific antibodies were detected in sera by ELISA. The virulent strain YPIII pIB1 (wild type) did not provoke polyclonal activation of B lymphocytes in vivo. In general, fewer Ig-secreting spleen cells of all isotypes were foundin the infected animals than in the control animals. Specific IgG antibodies were detected in the sera of animals infected with all strains. The peak response occurred on the 21st day post-infection, and the Yp100 strain provoked the highest level of these antibodies. We concluded that invasin is not a polyclonal activator of murine B cells.

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.

Prevalence and virulence properties of Vibrio cholerae non-O1, Aeromonas spp. and Plesiomonas shigelloides isolated from Cambé Stream (State of Paraná, Brazil).

The incidence of Vibrio cholerae, Aeromonas spp. and Plesiomonas shigelloides was determined in water samples from Cambé Stream. The samples were collected from seven different sites. The serogroups, virulence markers and drug resistance profiles were also evaluated. Twelve Aer. hydrophila, 12Aer. caviae, eight Aer. sobria, seven Ple. shigelloides and two V. cholerae non-O1 were isolated. They belonged to different serogroups and all produced haemolysis in different assays. Five of the Aeromonas strains and one of V cholerae non-O1 were positive for enterotoxin activity. Haemagglutination and its inhibition, using erythrocytes of different origins, was variable for Aeromonas spp. and V. cholerae, while none of the Ple. shigelloides haemagglutinated in association with any type of erythrocyte. All isolates exhibited multiple drug resistance. These results indicate that the occurrence of V. cholerae non-O1, Aeromonas spp. and Ple. shigelloides, in water used for vegetable irrigation, human recreation and animal consumption, among others, represents a potential risk for humans.

Characteristics of Yersinia pseudotuberculosis isolated from animals in Brazil.

Strains (105) of Yersinia pseudotuberculosis isolated in Brazil between 1982 and 1990 were bio-serotyped. They were also studied for plasmid profile, autoagglutination and calcium dependence at 37 degrees C, Congo red uptake, pyrazinamidase activity, esculin hydrolysis, salicin fermentation and drug sensitivity: 95.24% were biotype 2, serogroup O:3; 2.86% were biotype 1, serogroup O:1; and 1.90% were biotype 2, non-agglutinable. Plasmids were found in 77.14% of the strains (one in each strain). There was total correlation between the presence of the virulence plasmid and autoagglutination, calcium dependence at 37 degrees C and Congo red uptake. The esculin, salicin and pyrazinamidase tests were not efficient in differentiating pathogenic from non-pathogenic Y. pseudotuberculosis isolates. All strains were highly sensitive to the drugs used. These results indicate that Y. pseudotuberculosis is a potential pathogen for humans in Brazil, especially because the bio-serogroups detected among animals are those most frequently associated with human diseases.

Association between polyclonal B cell activation and the presence of autoantibodies in mice infected with Yersinia enterocolitica O:3.

Eight-week old conventional female Swiss mice were inoculated intravenously with Yersinia enterocolitica O:3. A second group of normal mice was used as control. Five mice from each group were bled by heart puncture and their spleens were removed for spleen cell collection on the 3rd, 5th, 7th, 10th, 14th and 21st day after infection. Immunoglobulin-secreting spleen cells were detected by the isotype-specific protein A plaque assay. Total immunoglobulin levels were determined in mouse serum by single radial immunodiffusion and the presence of autoantibodies was determined by ELISA. We observed a marked increase in the total number of cells secreting immunoglobulins of all isotypes as early as on the 3rd day post-infection and the peak of secretion occurred on the 7th day. At the peak of the immunoglobulin response, the total number of secreting cells was 19 times higher than that of control mice and most immunoglobulin-secreting cells were of the IgG2a isotype. On the 10th day post-infection, total serum immunoglobulin values were 2 times higher in infected animals when compared to the control group, and continued at this level up to the 21st day post-infection. Serum absorption with viable Y. enterocolitica cells had little effect on antibody levels detected by single radial immunodiffusion. Analysis of serum autoantibody levels revealed that Y. enterocolitica infection induced an increase of anti-myosin and anti-myelin immunoglobulins. The sera did not react with collagen. The present study demonstrates that Y. enterocolitica O:3 infection induces polyclonal activation of murine B cells which is correlated with the activation of some autoreactive lymphocyte clones.

Association between polyclonal B cell activation and the presence of autoantibodies in mice infected with Yersina enterocolitica O:3

Eight-week old conventional female Swiss mice were inoculated intravenously with Yersinia enterocolitica O:3. A second group of normal mice was used as control. Five mice from eaeh group were bled by heart puncture and their spleens were removed for spleen cell collection on the 3rd, 5th, 7th, 1Oth, l4th and 21st day after infection. Immunoglobulin-secreting spleen cells were detected by the isotypespecific protein A plaque assay. Total immunoglobulin levels were determined in mouse serum by single radial immunodiffusion and the presence of autoantibodies was determined by ELISA. We observed a marked increase in the total number of cells secreting immunoglobulins of all isotypes as early as on the 3rd day post-infection and the peak of secretion occurred on the 7th day. At the peak of the immunoglobulin response, the total number of secreting cells was 19 times higher than that of control mice and most immunoglobulin-secreting cells were of the IgG2a isotype. On the 10th day post-infection, total serum immunoglobulin values were 2 times higher in infected animals when compared to the control group, and continued at this level up to the 2lst day post-infection. Serum absorption with viable Y. enterocolitica cells had little effect on antibody levels detected by single radial immunodiffusion. Analysis of serum autoantibody levels revealed that Y. enterocolitica infection induced an increase of antimyosin and anti-myelin immunoglobulins. The sera did not react with collagen. The present study demonstrates that Y. enterocolitica O:3 infection induces polyclonal activation of murine B cells which is correlated with the activation of some autoreactive lymphocyte clones.

Microbiological quality of recreational waters in Araraquara, SP, Brazil.

The microbiological flora of 108 water samples was explored to evaluate the role of recreational waters as a possible source of human diseases in Araraquara, S.P., Brazil. These waters included six swimming pools and three lakes with beaches. The number of total and fecal coliforms, Escherichia coli, fecal streptococci, Pseudomonas aeruginosa, Staphylococcus aureus, Candida albicans, and heterotrophic organisms was determined. As was the occurrence of Salmonella, Shigella, yersinia, enteropathogenic E. coli (EPEC), enteroinvasive E. coli (EIEC), Mycobacteria, yeasts in general and dermatophytes. Shigella, Yersinia, EIEC and dermatophytes were not isolated. Other organisms or groups of microorganisms were found in variable proportions. From this study it is concluded that recreational waters used by the population of Araraquara, may be contaminated with potentially pathogenic microorganism and this may serve as a source of human diseases.

Lack of virulence factors in Escherichia coli strains of enteropathogenic serogroups isolated from water.

Thirty-eight Escherichia coli strains belonging to 14 human enteropathogenic serogroups were isolated from 33 of 208 water samples studied. No virulence factor or virulence-related gene sequences were found in any of the 38 strains analyzed. The results point out the importance of detecting specific virulence factors before incriminating water as a source of human diarrhea.

Occurrence of Yersinia spp. in foods in Brazil.

Over the past 9 years, 468 bacterial strains isolated from raw and pasteurized milk, beef and pork, bovine and chicken liver, chicken heart, gizzards and lung sausage, hamburger, cheese and lettuce in different regions of the State of São Paulo and in the city of Rio de Janeiro were received by the Reference Laboratory for Yersinia in Brazil. All were confirmed to be Yersinia spp. The 468 Yersinia isolates were grouped as 184 strains because some of the bacteria isolated from the same food sample belonged to the same species, and were considered to be a single strain. The Yersinia food strains were classified as Y. enterocolitica (46), Y. intermedia (67), Y. frederiksenii (20), Y. kristensenii (8) and 43 of them were biochemically atypical. Pathogenic types were not detected.