RESUMO
ABSTRACT The herein presented assay provided a bacteriological and molecular characterization of 100 samples of L. monocytogenes isolated from human (43) and food (57) sources, from several regions of Brazil, and collected between 1975 and 2013. Antigenic characterization defined 49% of serotype 4b samples, followed by 28% of serotype 1/2b, 14% of serotype 1/2c, 8% of serotype 1/2a, and 1% of serotype 3b. Both type of samples from human and food origin express the same serotype distribution. Multiplex PCR analysis showed 13 strains of type 4b with the amplification profile 4b-VI (Variant I). Virulence genes hly, inlA, inlB, inlC, inlJ, actA, plcA, and prfA were detected in all samples, highlighting a deletion of 105pb on the actA gene in 23% of serotype 4b samples. Macrorestriction profile with ApaI at PFGE showed 55 pulsotypes, with the occurrence of the same pulsotype in hospitalized patients in São Paulo in 1992 and 1997, and two other highly related pulsotypes in patients hospitalized in Rio de Janeiro in 2008. Recognized pulsotypes in listeriosis cases have also been detected in food. Thus, the prevalence of a serotype and the persistence of certain pulsotypes herald future problems.
Assuntos
Humanos , Fatores de Virulência/genética , Microbiologia de Alimentos , Listeria monocytogenes/genética , Brasil , Sorotipagem , Eletroforese em Gel de Campo Pulsado , Tipagem Molecular , Reação em Cadeia da Polimerase Multiplex , Genes Bacterianos/genética , Listeria monocytogenes/isolamento & purificação , Listeria monocytogenes/patogenicidadeRESUMO
The herein presented assay provided a bacteriological and molecular characterization of 100 samples of L. monocytogenes isolated from human (43) and food (57) sources, from several regions of Brazil, and collected between 1975 and 2013. Antigenic characterization defined 49% of serotype 4b samples, followed by 28% of serotype 1/2b, 14% of serotype 1/2c, 8% of serotype 1/2a, and 1% of serotype 3b. Both type of samples from human and food origin express the same serotype distribution. Multiplex PCR analysis showed 13 strains of type 4b with the amplification profile 4b-VI (Variant I). Virulence genes hly, inlA, inlB, inlC, inlJ, actA, plcA, and prfA were detected in all samples, highlighting a deletion of 105pb on the actA gene in 23% of serotype 4b samples. Macrorestriction profile with ApaI at PFGE showed 55 pulsotypes, with the occurrence of the same pulsotype in hospitalized patients in São Paulo in 1992 and 1997, and two other highly related pulsotypes in patients hospitalized in Rio de Janeiro in 2008. Recognized pulsotypes in listeriosis cases have also been detected in food. Thus, the prevalence of a serotype and the persistence of certain pulsotypes herald future problems.
Assuntos
Microbiologia de Alimentos , Listeria monocytogenes/genética , Fatores de Virulência/genética , Brasil , Eletroforese em Gel de Campo Pulsado , Genes Bacterianos/genética , Humanos , Listeria monocytogenes/isolamento & purificação , Listeria monocytogenes/patogenicidade , Tipagem Molecular , Reação em Cadeia da Polimerase Multiplex , SorotipagemRESUMO
We sought to characterize pneumococcal isolates associated with bacteremia, pneumonia and meningitis in cancer patients and to estimate the coverage of the available pneumococcal vaccines. Fifty isolates recovered from 49 patients attending a cancer reference center over a 1-year period were analyzed. The prevalent serotypes were: 23F (12%), 6A (8%), 3, 4, 20, and 23A (6% each). All isolates were susceptible to chloramphenicol, levofloxacin, rifampicin, and vancomycin. Resistance or reduced susceptibility to penicillin made up 14%, and one isolate was also intermediately resistant to ceftriaxone. The three (6%) erythromycin-resistant isolates presented the M or cMLSB phenotypes and harbored the mef(A/E) gene exclusively or along with the erm(B) gene. Twenty-two (44%) isolates were closely related to 11 international clones, being strongly associated with penicillin non-susceptibility. Combined immunization with the 13-valent conjugate and the 23-valent polysaccharide vaccines might contribute to reduce (76%) the burden of the pneumococcal infections in the population investigated.
Assuntos
Antibacterianos/uso terapêutico , Farmacorresistência Bacteriana Múltipla/genética , Infecções Pneumocócicas/tratamento farmacológico , Vacinas Pneumocócicas/imunologia , Streptococcus pneumoniae/imunologia , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Bacteriemia/tratamento farmacológico , Bacteriemia/microbiologia , Brasil , Criança , Pré-Escolar , Feminino , Humanos , Lactente , Masculino , Meningites Bacterianas/tratamento farmacológico , Meningites Bacterianas/microbiologia , Testes de Sensibilidade Microbiana , Pessoa de Meia-Idade , Neoplasias/microbiologia , Pneumonia/tratamento farmacológico , Pneumonia/microbiologia , Sorogrupo , Streptococcus pneumoniae/classificação , Streptococcus pneumoniae/genética , Streptococcus pneumoniae/isolamento & purificação , Adulto JovemRESUMO
Listeria spp. isolated from different food products and collected from 12 Brazilian states were sent to the Laboratory of Bacterial Zoonoses (Oswaldo Cruz Institute, Brazil) for identification. The aims of this study were to characterize these isolates, from 1990 to 2012, by using biochemical, morphological, and serotyping tests, and to analyze the distribution of L. monocytogenes serotypes on different food products and geographical locations. Serotyping was performed using polyclonal somatic and flagellar antisera. Of 5953 isolates, 5770 were identified as Listeria spp., from which 3429 (59.4%) were L. innocua, 2248 (38.9%) were L. monocytogenes, and 93 (1.6%) were other Listeria spp. L. innocua was predominantly isolated from 1990 to 2000, while L. monocytogenes was from 2001 to 2012. Regarding the serotype distribution in the foods, serotypes 1/2a and 4b were most common in processed meat and ready-to-eat products, respectively; serotypes 1/2a, 1/2b, and 4b were the most common in nonprocessed meat. The results above confirm the presence of the main serotypes of L. monocytogenes in different parts of the food chain from three regions of the country and emphasize the importance of improving the control measures, as tolerance zero policy and microbiological surveillance in Brazil.
Assuntos
Microbiologia de Alimentos , Listeria monocytogenes/isolamento & purificação , Listeriose/genética , Sorogrupo , Brasil , Contagem de Colônia Microbiana , Humanos , Listeria monocytogenes/genética , Listeria monocytogenes/patogenicidade , Listeriose/epidemiologia , Listeriose/microbiologia , Carne/microbiologia , Sorotipagem/métodosRESUMO
Poultry are considered to be the main reservoir of Campylobacterspp. bacteria, an important pathogen for humans. Many studies have reported a rapid selection of fluoroquinolone-resistant strains following the widespread use of these antimicrobials in poultry production and human medicine. The main mechanism of fluoroquinolone resistance in Campylobacteris a mutation in the Quinolone Resistance Determinant Region (QRDR) in the gyrA gene, which codes for the subunit of the enzyme DNA gyrase, the target for fluoroquinolone. The aim of this study was to investigate the mutation in QRDR in the gyrA gene of Campylobacterstrains previously isolated from broiler carcasses and feces of laying hens. Thirty-eight strains of C. jejuniand 19 C. colistrains (n=57), previously characterized as resistant to ciprofloxacin and enrofloxacin by the disk diffusion method and minimum inhibitory concentration (MIC), were selected. For detection of the mutation, a fragment of 454pb QRDR in the gyrA gene was used for direct sequencing. All strains presented the QRDR mutation in the gyrA gene at codon 86 (Thr-86-Ile), which confers resistance to fluoroquinolones. Other known silent mutations were observed. This genotypic characterization of fluoroquinolone resistance inCampylobacterstrains has confirmed the prior phenotypic detection of the resistance. The Thr-86-Ile mutation was observed in all samples confirming that this is the predominant mutation in enrofloxacin and ciprofloxacin resistant strains of C. jejuniand C. coli.
As aves são consideradas o principal reservatório deCampylobacterspp., um importante patógeno para humanos e muitos estudos têm relatado uma rápida seleção de cepas resistentes às fluoroquinolonas após o uso destes antimicrobianos na produção avícola e na medicina humana. O principal mecanismo de resistência às fluoroquinolonas em Campylobacterconsiste na mutação na Região Determinantes de Resistência às Quinolonas (RDRQ) do gene gyrA, que codifica para a subunidade A da enzima DNA girase, alvo das fluoroquinolonas. O objetivo deste estudo foi investigar a mutação na RDRQ do gene gyrA em cepas de Campylobacterpreviamente isolados de carcaças de frangos de corte e fezes de galinhas poedeiras. Foram selecionadas 38 cepas de C. jejunie 19 cepas de C. coli(n=57), previamente caracterizadas como resistentes à ciprofloxacina e enrofloxacina, pelo método da difusão em disco e pela determinação da concentração inibitória mínima. Para detecção da mutação, foi utilizado sequenciamento direto de um fragmento de 454pb da RDRQ do gene gyrA gerado por PCR. Todas as cepas apresentaram a mutação na RDRQ do gene gyrA no códon 86 (Tre-86-Ile), que confere resistência às fluoroquinolonas e outras mutações silenciosas foram observadas. A caracterização genotípica da resistência às fluoroquinolonas em Campylobacterconfirmou a prévia detecção fenotípica dessa resistência e a mutação Tre-86-Ile foi observada na totalidade das amostras, comprovando ser esta a mutação predominante em cepas de C. jejunie C. coliresistentes à enrofloxacina e ciprofloxacina.
RESUMO
As aves são consideradas o principal reservatório de Campylobacter spp., um importante patógeno para humanos e muitos estudos têm relatado uma rápida seleção de cepas resistentes às fluoroquinolonas após o uso destes antimicrobianos na produção avícola e na medicina humana. O principal mecanismo de resistência às fluoroquinolonas em Campylobacter consiste na mutação na Região Determinantes de Resistência às Quinolonas (RDRQ) do gene gyrA, que codifica para a subunidade A da enzima DNA girase, alvo das fluoroquinolonas. O objetivo deste estudo foi investigar a mutação na RDRQ do gene gyrA em cepas de Campylobacter previamente isolados de carcaças de frangos de corte e fezes de galinhas poedeiras. Foram selecionadas 38 cepas de C. jejuni e 19 cepas de C. coli (n=57), previamente caracterizadas como resistentes à ciprofloxacina e enrofloxacina, pelo método da difusão em disco e pela determinação da concentração inibitória mínima. Para detecção da mutação, foi utilizado sequenciamento direto de um fragmento de 454pb da RDRQ do gene gyrA gerado por PCR. Todas as cepas apresentaram a mutação na RDRQ do gene gyrA no códon 86 (Tre-86-Ile), que confere resistência às fluoroquinolonas e outras mutações silenciosas foram observadas. A caracterização genotípica da resistência às fluoroquinolonas em Campylobacter confirmou a prévia detecção fenotípica dessa resistência e a mutação Tre-86-Ile foi observada na totalidade das amostras, comprovando ser esta a mutação predominante em cepas de C. jejuni e C. coli resistentes à enrofloxacina e ciprofloxacina.(AU)
Poultry are considered to be the main reservoir of Campylobacter spp. bacteria, an important pathogen for humans. Many studies have reported a rapid selection of fluoroquinolone-resistant strains following the widespread use of these antimicrobials in poultry production and human medicine. The main mechanism of fluoroquinolone resistance in Campylobacter is a mutation in the Quinolone Resistance Determinant Region (QRDR) in the gyrA gene, which codes for the subunit of the enzyme DNA gyrase, the target for fluoroquinolone. The aim of this study was to investigate the mutation in QRDR in the gyrA gene of Campylobacter strains previously isolated from broiler carcasses and feces of laying hens. Thirty-eight strains of C. jejuni and 19 C. coli strains (n=57), previously characterized as resistant to ciprofloxacin and enrofloxacin by the disk diffusion method and minimum inhibitory concentration (MIC), were selected. For detection of the mutation, a fragment of 454pb QRDR in the gyrA gene was used for direct sequencing. All strains presented the QRDR mutation in the gyrA gene at codon 86 (Thr-86-Ile), which confers resistance to fluoroquinolones. Other known silent mutations were observed. This genotypic characterization of fluoroquinolone resistance in Campylobacter strains has confirmed the prior phenotypic detection of the resistance. The Thr-86-Ile mutation was observed in all samples confirming that this is the predominant mutation in enrofloxacin and ciprofloxacin resistant strains of C. jejuni and C. coli.(AU)
Assuntos
Fluoroquinolonas , DNA Girase , Campylobacter/isolamento & purificação , Aves Domésticas , MutaçãoRESUMO
INTRODUCTION: Listeria monocytogenes is an important foodborne pathogen and the 4b serotype is responsible for many cases of human listeriosis reported in Brazil. Several listeriosis outbreaks worldwide have involved a small number of well-defined clonal groups, designated as epidemic clones (ECs). METHODOLOGY: We studied 71 strains of serotype 4b, including 25 isolates from human cases of listeriosis and 46 from meat-based foods, collected in Brazil between 1977 and 2010. The presence of ECs (I and II) markers and virulence genes (inlA, inlB, ilnC, inlJ and actA) were evaluated by PCR assay. The genetic relationship of ECs-positive strains was assessed by pulsed field gel electrophoresis. RESULTS: ECI and ECII markers were found both in human and food strains, with 19.7% positive for the ECI marker and 40.8% for ECII. Most strains (97.2%) were positive for the virulence genes that were studied. Nevertheless, the actA gene amplicons showed two distinct sizes, with all ECI positive strains exhibiting a 105bp deletion. Pulsed field gel electrophoresis (PFGE) analysis allowed the recognition of highly related strains, particularly from two outbreaks of neonatal listeriosis in São Paulo State occurred in 1992 and 1997, both ECII-positive; and two ECI strains from a human case (1982) and from bovine meat (2009). CONCLUSIONS: The presence of ECs among clinical samples and beef isolates of serotype 4b from some regions of Brazil highlights the need for rigorous control of production procedures. Furthermore, the association of ECII with two nosocomial outbreaks suggests its ability to spread in these settings.
Assuntos
Genótipo , Listeria monocytogenes/classificação , Listeria monocytogenes/isolamento & purificação , Listeriose/microbiologia , Produtos da Carne/microbiologia , Sorogrupo , Animais , Brasil/epidemiologia , Bovinos , Eletroforese em Gel de Campo Pulsado , Marcadores Genéticos , Humanos , Listeria monocytogenes/genética , Listeriose/epidemiologia , Tipagem Molecular , Reação em Cadeia da Polimerase , Fatores de Virulência/genéticaRESUMO
Neste estudo, são apresentados os resultados obtidos em uma pesquisa para bactérias entéricas conduzida entre os Xavantes, grupo indígena de Mato Grosso, Brasil, a partir de fezes conservadas em meio de transporte. Os resultados mostraram, por meio datécnica clássica de isolamento, bioquímica e sorologia, a presença de duas espécies bacterianas importantes, causadoras de diarreia: Salmonella enterica e Escherichia coli enteropatogênica (EPEC). Essas mesmas amostras, pesquisadas para outros agentes não bacterianos, indicaram também, em 40% dos casos, associação com parasitos, sugerindo uma relação direta com a baixa salubridade da comunidade e a necessidade da implementação de saneamento básico...
Assuntos
Humanos , Diarreia , Escherichia coli Enteropatogênica , Povos Indígenas , Salmonella entericaRESUMO
INTRODUCTION: Yersinia enterocolitica is a well-known foodborne pathogen widely distributed in nature with high public health relevance, especially in Europe. METHODOLOGY: This study aimed to analyze the pathogenic potential of Y. enterocolitica isolated strains from human, animal, food, and environmental sources and from different regions of Brazil by detecting virulence genes inv, ail, ystA, and virF through polymerase chain reaction (PCR), phenotypic tests, and antimicrobial susceptibility analysis. Pulsed-field gel electrophoresis (PFGE) was used for the assessment of phylogenetic diversity. RESULTS: All virulence genes were detected in 11/60 (18%) strains of serotype O:3, biotype 4 isolated from human and animal sources. Ten human strains (4/O:3) presented three chromosomal virulence genes, and nine strains of biotype 1A presented the inv gene. Six (10%) strains were resistant to sulfamethoxazole-trimethoprim, seven (12%) to tetracycline, and one (2%) to amikacin, all of which are used to treat yersiniosis. AMP-CEF-SXT was the predominant resistance profile. PFGE analysis revealed 36 unique pulsotypes, grouped into nine clusters (A to I) with similarity ≥ 85%, generating a diversity discriminatory index of 0.957. Cluster A comprised all bio-serotype 4/O:3 strains isolated from animal and humans sources. CONCLUSIONS: This study shows the existence of strains with the same genotypic profiles, bearing all virulence genes, from human and animal sources, circulating among several Brazilian states. This supports the hypothesis that swine is likely to serve as a main element in Y. enterocolitica transmission to humans in Brazil, and it could become a potential threat to public health as in Europe.
Assuntos
Yersinia enterocolitica/classificação , Yersinia enterocolitica/isolamento & purificação , Animais , Antibacterianos/farmacologia , Técnicas de Tipagem Bacteriana , Brasil/epidemiologia , Análise por Conglomerados , DNA Bacteriano/genética , Eletroforese em Gel de Campo Pulsado , Microbiologia Ambiental , Microbiologia de Alimentos , Genes Bacterianos , Variação Genética , Genótipo , Humanos , Testes de Sensibilidade Microbiana , Reação em Cadeia da Polimerase , Sorogrupo , Suínos , Fatores de Virulência/genética , Yersiniose/epidemiologia , Yersiniose/microbiologia , Yersiniose/veterinária , Yersinia enterocolitica/genética , Yersinia enterocolitica/fisiologia , Zoonoses/epidemiologia , Zoonoses/microbiologiaRESUMO
Cantagalo virus is a strain of vaccinia virus (genus Orthopoxvirus) and the etiological agent of an important vesicopustular disease that affects dairy cows and milkers in Brazil. The reservoirs involved in the maintenance of this virus in nature are unknown. In the present work, the detection of neutralizing antibodies to Orthopoxvirus in capybaras collected in São Paulo state is reported. Capybaras are the largest rodent species native to South America and have already been reported as putative reservoirs of other pathogenic microorganisms. Thirteen out of thirty-three serum samples were found positive in plaque-reduction neutralization tests, some of them showing high titers compared to positive controls. These results suggest that capybaras may play a role in the infection cycle of vaccinia virus in Brazil.