RESUMO
AIM: The occurrence of virulence markers, serotypes and invasive ability were investigated in Shiga toxin-producing Escherichia coli (STEC) isolated from faecal samples of healthy dairy cattle at Rio de Janeiro State, Brazil. METHODS AND RESULTS: From 1562 stx-positive faecal samples, 105 STEC strains were isolated by immuno-magnetic separation (IMS) or plating onto MacConkey agar (MC) followed by colony hybridisation. Fifty (47·6%) strains belonged to nine serotypes (O8:H19, O22:H8, O22:H16, O74:H42, O113:H21, O141:H21, O157:H7, O171:H2 and ONT:H21). The prevalent serotypes were O157:H7 (12·4%), O113:H21 (6·7%) and O8:H19 (5·7%). Virulence genes were identified by polymerase chain reaction (PCR). E-hlyA (77·1%) was the more prevalent virulence marker, followed by espP (64·8%), saa (39%), eae (24·8%) and astA (21·9%). All O157:H7 strains carried the γ (gamma) variant of the locus of enterocyte effacement (LEE) genes and the stx2c gene, while the stx1/stx2 genotype prevailed among the eae-negative strains. None of the eae-positive STEC produced the localized adherence (LA) phenotype in HEp-2 or Caco-2 cells. However, intimate attachment (judged by the fluorescent actin staining test) was detected in some eae-positive strains, both in HEp-2 (23·1%) and in Caco-2 cells (11·5%). Most strains (87·5%) showed 'peripheral association' (PA) adherence phenotype to undifferentiated Caco-2 cells. Twenty-five (92·6%) of 27 strains invaded Caco-2 cells. The highest average value of invasion (9·6%) was observed among the eae-negative bovine strains from serotypes described in human disease. CONCLUSION: Healthy dairy cattle is a reservoir of STEC carrying virulence genes and properties associated with human disease. SIGNIFICANCE AND IMPACT OF THE STUDY: Although reports of human disease associated with STEC are scarce in Brazil, the colonization of the animal reservoir by potentially pathogenic strains offers a significant risk to our population.
Assuntos
Bovinos/microbiologia , Reservatórios de Doenças/microbiologia , Escherichia coli Shiga Toxigênica/isolamento & purificação , Animais , Brasil , Células CACO-2 , Proteínas de Escherichia coli/genética , Proteínas de Escherichia coli/metabolismo , Fezes/microbiologia , Humanos , Reação em Cadeia da Polimerase , Sorotipagem , Toxina Shiga/metabolismo , Escherichia coli Shiga Toxigênica/classificação , Escherichia coli Shiga Toxigênica/genética , Escherichia coli Shiga Toxigênica/fisiologia , Virulência/genética , Fatores de Virulência/genética , Fatores de Virulência/metabolismoRESUMO
Variation in annual rainfall is considered the most important factor influencing population dynamics in dry environments. However, different factors may control population dynamics in different microhabitats. This study recognizes that microhabitat variation may attenuate the influence of climatic seasonality on the population dynamics of herbaceous species in dry forest (Caatinga) areas of Brazil. We evaluated the influence of three microhabitats (flat, rocky and riparian) on the population dynamics of four herbaceous species (Delilia biflora, Commelina obliqua, Phaseolus peduncularis and Euphorbia heterophylla) in a Caatinga (dry forest) fragment at the Experimental Station of the Agronomic Research Institute of Pernambuco in Brazil, over a period of three years. D. biflora, C. obliqua and P. peduncularis were found in all microhabitats, but they were present at low densities in the riparian microhabitat. There was no record of E. heterophylla in the riparian microhabitat. Population size, mortality rates and natality rates varied over time in each microhabitat. This study indicates that different establishment conditions influenced the population size and occurrence of the four species, and it confirms that microhabitat can attenuate the effect of drought stress on mortality during the dry season, but the strength of this attenuator role may vary with time and species.
Assuntos
Clima Desértico , Florestas , Magnoliopsida/fisiologia , Brasil , Densidade Demográfica , Estações do Ano , Especificidade da EspécieRESUMO
Abstract Variation in annual rainfall is considered the most important factor influencing population dynamics in dry environments. However, different factors may control population dynamics in different microhabitats. This study recognizes that microhabitat variation may attenuate the influence of climatic seasonality on the population dynamics of herbaceous species in dry forest (Caatinga) areas of Brazil. We evaluated the influence of three microhabitats (flat, rocky and riparian) on the population dynamics of four herbaceous species (Delilia biflora, Commelina obliqua, Phaseolus peduncularis and Euphorbia heterophylla) in a Caatinga (dry forest) fragment at the Experimental Station of the Agronomic Research Institute of Pernambuco in Brazil, over a period of three years. D. biflora, C. obliqua and P. peduncularis were found in all microhabitats, but they were present at low densities in the riparian microhabitat. There was no record of E. heterophylla in the riparian microhabitat. Population size, mortality rates and natality rates varied over time in each microhabitat. This study indicates that different establishment conditions influenced the population size and occurrence of the four species, and it confirms that microhabitat can attenuate the effect of drought stress on mortality during the dry season, but the strength of this attenuator role may vary with time and species.(AU)
Resumo A variação anual na precipitação é considerada o fator mais importante que influencia a dinâmica populacional em ambientes secos. No entanto, diferentes fatores podem controlar a dinâmica populacional em diferentes microhabitats. A hipótese deste estudo é que as variações de microhabitats podem atenuar a influência da sazonalidade climática sobre a dinâmica populacional de espécies herbáceas em uma áreas floresta seca (Caatinga) no Brasil. Então, objetivou-se avaliar a influência de três microhabitats (plano, rochoso e ciliar) sobre a dinâmica das populações de quatro espécies herbáceas (Delilia biflora, Commelina obliqua, Phaseolus peduncularis e Euphorbia heterophylla) em um fragmento de Caatinga da Estação Experimental do Instituto de Pesquisas Agronômicas de Pernambuco, no Brasil, durante um período de três anos. D. biflora, C. obliqua e P. peduncularis foram encontrados em todos os microhabitats, mas elas estavam presentes em baixas densidades no microhabitat ciliar. Não houve registro de E. heterophylla no microhabitat ciliar. O tamanho das populações, as taxas de mortalidade e de natalidade variaram ao longo do tempo em cada microhabitat. Este estudo indica que diferentes as condições de estabelecimento influenciaram o tamanho das populações e a ocorrência das quatro espécies e confirma que microhabitat pode atenuar o efeito do estresse hídrico sobre a mortalidade durante a estação seca, mas a força desse papel atenuador pode variar com o tempo e a espécie considerada.(AU)
Assuntos
Ecossistema , Análise Espacial , Zona Semiárida , DemografiaRESUMO
Abstract Variation in annual rainfall is considered the most important factor influencing population dynamics in dry environments. However, different factors may control population dynamics in different microhabitats. This study recognizes that microhabitat variation may attenuate the influence of climatic seasonality on the population dynamics of herbaceous species in dry forest (Caatinga) areas of Brazil. We evaluated the influence of three microhabitats (flat, rocky and riparian) on the population dynamics of four herbaceous species (Delilia biflora, Commelina obliqua, Phaseolus peduncularis and Euphorbia heterophylla) in a Caatinga (dry forest) fragment at the Experimental Station of the Agronomic Research Institute of Pernambuco in Brazil, over a period of three years. D. biflora, C. obliqua and P. peduncularis were found in all microhabitats, but they were present at low densities in the riparian microhabitat. There was no record of E. heterophylla in the riparian microhabitat. Population size, mortality rates and natality rates varied over time in each microhabitat. This study indicates that different establishment conditions influenced the population size and occurrence of the four species, and it confirms that microhabitat can attenuate the effect of drought stress on mortality during the dry season, but the strength of this attenuator role may vary with time and species.
Resumo A variação anual na precipitação é considerada o fator mais importante que influencia a dinâmica populacional em ambientes secos. No entanto, diferentes fatores podem controlar a dinâmica populacional em diferentes microhabitats. A hipótese deste estudo é que as variações de microhabitats podem atenuar a influência da sazonalidade climática sobre a dinâmica populacional de espécies herbáceas em uma áreas floresta seca (Caatinga) no Brasil. Então, objetivou-se avaliar a influência de três microhabitats (plano, rochoso e ciliar) sobre a dinâmica das populações de quatro espécies herbáceas (Delilia biflora, Commelina obliqua, Phaseolus peduncularis e Euphorbia heterophylla) em um fragmento de Caatinga da Estação Experimental do Instituto de Pesquisas Agronômicas de Pernambuco, no Brasil, durante um período de três anos. D. biflora, C. obliqua e P. peduncularis foram encontrados em todos os microhabitats, mas elas estavam presentes em baixas densidades no microhabitat ciliar. Não houve registro de E. heterophylla no microhabitat ciliar. O tamanho das populações, as taxas de mortalidade e de natalidade variaram ao longo do tempo em cada microhabitat. Este estudo indica que diferentes as condições de estabelecimento influenciaram o tamanho das populações e a ocorrência das quatro espécies e confirma que microhabitat pode atenuar o efeito do estresse hídrico sobre a mortalidade durante a estação seca, mas a força desse papel atenuador pode variar com o tempo e a espécie considerada.
Assuntos
Magnoliopsida/fisiologia , Clima Desértico , Florestas , Brasil , Densidade Demográfica , Estações do Ano , Especificidade da EspécieRESUMO
Abstract Variation in annual rainfall is considered the most important factor influencing population dynamics in dry environments. However, different factors may control population dynamics in different microhabitats. This study recognizes that microhabitat variation may attenuate the influence of climatic seasonality on the population dynamics of herbaceous species in dry forest (Caatinga) areas of Brazil. We evaluated the influence of three microhabitats (flat, rocky and riparian) on the population dynamics of four herbaceous species (Delilia biflora, Commelina obliqua, Phaseolus peduncularis and Euphorbia heterophylla) in a Caatinga (dry forest) fragment at the Experimental Station of the Agronomic Research Institute of Pernambuco in Brazil, over a period of three years. D. biflora, C. obliqua and P. peduncularis were found in all microhabitats, but they were present at low densities in the riparian microhabitat. There was no record of E. heterophylla in the riparian microhabitat. Population size, mortality rates and natality rates varied over time in each microhabitat. This study indicates that different establishment conditions influenced the population size and occurrence of the four species, and it confirms that microhabitat can attenuate the effect of drought stress on mortality during the dry season, but the strength of this attenuator role may vary with time and species.
Resumo A variação anual na precipitação é considerada o fator mais importante que influencia a dinâmica populacional em ambientes secos. No entanto, diferentes fatores podem controlar a dinâmica populacional em diferentes microhabitats. A hipótese deste estudo é que as variações de microhabitats podem atenuar a influência da sazonalidade climática sobre a dinâmica populacional de espécies herbáceas em uma áreas floresta seca (Caatinga) no Brasil. Então, objetivou-se avaliar a influência de três microhabitats (plano, rochoso e ciliar) sobre a dinâmica das populações de quatro espécies herbáceas (Delilia biflora, Commelina obliqua, Phaseolus peduncularis e Euphorbia heterophylla) em um fragmento de Caatinga da Estação Experimental do Instituto de Pesquisas Agronômicas de Pernambuco, no Brasil, durante um período de três anos. D. biflora, C. obliqua e P. peduncularis foram encontrados em todos os microhabitats, mas elas estavam presentes em baixas densidades no microhabitat ciliar. Não houve registro de E. heterophylla no microhabitat ciliar. O tamanho das populações, as taxas de mortalidade e de natalidade variaram ao longo do tempo em cada microhabitat. Este estudo indica que diferentes as condições de estabelecimento influenciaram o tamanho das populações e a ocorrência das quatro espécies e confirma que microhabitat pode atenuar o efeito do estresse hídrico sobre a mortalidade durante a estação seca, mas a força desse papel atenuador pode variar com o tempo e a espécie considerada.
RESUMO
BACKGROUND: Chronic hepatitis B (CHB) and schistosomiasis are prevalent in several countries, but the impact of this association is unknown. We aimed to investigate the prevalence and morbidity of this co-infection in Minas Gerais, an endemic area of schistosomiasis in Brazil. METHODS: In total, 406 adults with CHB (HBsAg positive >6 months) were included in a cross-sectional study. CHB was classified as replicative (HBV DNA ≥ 2.000 IU/ml), and low replicative or inactive hepatitis B carriers (HBV DNA <2.000 IU/ml). Schistosomiasis was confirmed by epidemiological and clinical records. Liver biopsies were scored by METAVIR. The risk of severe fibrosis was estimated by multivariate analysis. RESULTS: Of the 406 patients, 64.8% (263) were male, and the median age was 45 years (IQR 35-54). In total, 57.9% (235) had replicative CHB, and 31.5% (128) had cirrhosis. Schistosoma mansoni was confirmed in 30.5% (124) patients, 81.5% (101) of which were male with a median age of 47 years (IQR 39.5-54). Of the co-infected patients, 61.3% (76) and 38.7% (48) had replicative and inactive CHB, respectively. Schistosomal portal fibrosis (PF) was detected in 69.4% (86/124) patients. Patients with replicative CHB and schistosomal PF had more advanced fibrosis and severe inflammation compared with patients without schistosomal PF (80.8% vs 43.6% for METAVIR F3-F4, p<0.01; 64.0% vs 39.8% for METAVIR A2-A3, p < 0.01). Age >50 years (OR = 1.10; 95% CI 1.06-1.14, p<0.001), male gender (OR = 2.61, 95% CI 1.12-6.09, p = 0.03), schistosomal PF (OR = 4.56, 95% CI 2.10-9.91, p<0.001) and alcoholism (OR = 2.46, 95% CI 1.16-5.19, p = 0.02) were independently associated with cirrhosis. CONCLUSIONS: The association between replicative CHB and schistosomal PF can be a risk factor for more severe liver disease, which can result in deleterious outcomes for patients from endemic areas.
Assuntos
Hepatite B Crônica/epidemiologia , Esquistossomose mansoni/epidemiologia , Adulto , Distribuição por Idade , Brasil/epidemiologia , Comorbidade , Estudos Transversais , Feminino , Humanos , Fígado/parasitologia , Cirrose Hepática/epidemiologia , Cirrose Hepática/etiologia , Masculino , Pessoa de Meia-Idade , Análise Multivariada , Prevalência , Distribuição por SexoRESUMO
The study was undertaken to determine the clonal relationship and the genetic diversity among Escherichia coli isolates by comparing a non-motile O157 variant with three O157:H7 EHEC isolates and one O55:H7 enteropathogenic E. coli (EPEC) strain. E. coli strains were characterized by sorbitol phenotype, multilocus enzyme electrophoresis, pulsed-field gel electrophoresis, random amplification polymorphic DNA, and the presence of specific virulence genes (stx, E-hly and LEE genes). Sorbitol fermentation was observed in O157:H- (strain 116I), O55:H7 and O157:H7 (strain GC148) serotypes. stx1 or stx2 and E-hly genes were only detected among O157:H7 isolates. LEE typing revealed specific allele distribution: eaegamma, tirgamma, espAgamma, espBgamma associated with EPEC O55:H7 and EHEC O157:H7 strains (B1/1 and EDL 933), eaealpha, tiralpha, espAalpha, espBalpha related to the 116I O157:H- strain and the GC148 strain presented non-typable LEE sequences. Multilocus enzyme profiles revealed two main clusters associated with specific LEE pathotypes. E. coli strains were discriminated by random amplification of polymorphic DNA-polymerase chain reaction and pulsed-field gel electrophoresis methodologies. The molecular approaches used in this study allowed the determination of the genetic relatedness among E. coli strains as well as the detection of lineage specific group markers.
Assuntos
Escherichia coli Enteropatogênica/classificação , Escherichia coli Enteropatogênica/genética , Escherichia coli O157/classificação , Escherichia coli O157/genética , Animais , Técnicas de Tipagem Bacteriana , Bovinos , Linhagem Celular , Impressões Digitais de DNA , DNA Bacteriano/genética , Eletroforese em Gel de Campo Pulsado , Escherichia coli Enteropatogênica/enzimologia , Infecções por Escherichia coli/microbiologia , Infecções por Escherichia coli/veterinária , Escherichia coli O157/enzimologia , Fermentação , Humanos , Técnica de Amplificação ao Acaso de DNA Polimórfico , Sorbitol/metabolismo , Fatores de Virulência/genéticaRESUMO
Fifty-six Escherichia coli strains, serogrouped as EPEC, isolated from three different brands of pasteurised milk commercialised in Rio de Janeiro, Brazil, were tested for enteropathogenicity markers. Most of the strains (71.4%) were adherent to HEp-2 cells. The adherent strains were distributed among 7 EPEC serogroups (O26, O55, O111, O114, O125, O127, O128, O158). Although almost half of these strains (33.9%) presented unrecognisable adherence phenotypes, classical adherence patterns (localised-like, aggregative and diffuse adherence) described for E. coli and epidemiologically associated with diarrheagenic strains were observed. None of the strains showed typical localised adherence, usually associated with EPEC strains, but 4 of them displayed a localised-like adherence (LAL) phenotype, characterised by fewer and less compact microcolonies but that is still associated with diarrheagenic strains as well as strains of non-human origin. Indeed, 3 of these 4 strains were able to elicit the attaching-effacing lesion (FAS-positive), the central feature of EPEC pathogenesis, and hybridised with bfpA and eae DNA probes. The other LAL-positive strain hybridised with the bfpA probe but gave negative results for the eae probe and FAS assays. Interestingly, all LAL-positive strains produced amplicons of 200 bp in the PCR for bfpA, instead of the expected 326 bp fragment. PCR reactions for stx1 and stx2, two shiga-toxin-encoding genes, gave negative results. Typing of LEE-associated genes by PCR showed the profile eae (beta), tir (beta), espA (alpha) and espB (alpha) for one of the LAL-positive strain. The most prevalent adherence phenotype was the aggregative pattern which is observed in strains epidemiologically associated with persistent diarrhea. Additionally, one strain promoted complete detachment of the Hep-2 cell monolayer after 3 h of infection which might be related to the production of citotoxins, a feature that has been increasingly observed in clinical strains. The possession of EPEC-related O and H antigens is no longer deemed an essential characteristic of true pathogenic EPEC strains, emphasising the importance of routinely screen for virulence markers in E. coli strains isolated from foods. Our results are in accordance with data from the literature that demonstrate that environmental strains display atypical features but yet are capable of eliciting the classical A/E lesion and thus must be considered as potentially pathogenic. Further, our results demonstrate the potential of pasteurised milk as a vehicle for transmission of diarrheagenic E. coli in Brazil.
Assuntos
Proteínas de Escherichia coli , Escherichia coli/classificação , Escherichia coli/patogenicidade , Microbiologia de Alimentos , Leite/microbiologia , Toxinas Shiga/biossíntese , Animais , Aderência Bacteriana , Sequência de Bases , Linhagem Celular , Qualidade de Produtos para o Consumidor , Sondas de DNA , Escherichia coli/fisiologia , Genótipo , Humanos , Fenótipo , Filogenia , Sorotipagem , VirulênciaRESUMO
No período de março 2001 a julho 2002 foram estudadas 2823 fêmeas bovinas, em idade de reprodução, mestiças das raças Holandesa e Gir, pertencentes a 34 propriedades da bacia leiteira de Goiânia. A taxa de prenhez foi 47,8%. Entre os animais não gestantes (n= 1473), 17% apresentaram alterações inflamatórias do útero. Outras anormalidades encontradas foram: aplasia parcial do sistema genital (0,04%), feto macerado (0,01%), ovário aderido (0,04%). Casos de natimortalidade (0,04%), retenção placentária (0,01%) e cervicite (0,6%) e taxas de abortos (0,88%) também foram registrados. Cocos Gram positivos (41,3%) e bastonetes Gram negativos (52,6%) foram os microrganismos mais isolados, sendo Staphylococcus aureus e Escherichia coli os principais patógenos encontrados nas infeccções uterinas. Testes in vitro mostraram que esses microrganismos apresentaram maior susceptibilidade frente ao cloranfenicol, à gentamicina e à neomicina.(AU)
An epidemiological study was carried out on 2823 cows from 34 dairy herds from Goiania in the State of Goias-Brazil during 2001 to 2002. The pregnancy rate was 47.8%. In 1473 non-pregnant cows, causes of reproductive failure problems were sought. The most prevalent uterine infection was endometritis (17.0%). Uterine disorders such as partial hypoplasia of the genital system (0.04%), macerate fetus (0.01%), adhesion of ovaries (0.04%), stillbirth (0.04%), retained placenta (0.01%), cervix inflammation (0.6%) and abortion (0.88%) also were found. Uterine swabs were collected aseptically for microbiological culture. Gram positives cocci (41.3%) and Gram negatives rods (52.6%) were found, and Staphylococcus aureus and Escherichia coli were the most prevalent pathogens. Susceptibility patterns of microorganisms suggested the use of chloramphenicol, gentamicin and neomycin for antimicrobial therapy.(AU)
Assuntos
Endometrite/epidemiologia , Endometrite/etiologia , Staphylococcus aureus/isolamento & purificação , Escherichia coli/isolamento & purificação , Testes de Sensibilidade Microbiana/métodos , BovinosRESUMO
No período de março 2001 a julho 2002 foram estudadas 2823 fêmeas bovinas, em idade de reprodução, mestiças das raças Holandesa e Gir, pertencentes a 34 propriedades da bacia leiteira de Goiânia. A taxa de prenhez foi 47,8%. Entre os animais não gestantes (n= 1473), 17% apresentaram alterações inflamatórias do útero. Outras anormalidades encontradas foram: aplasia parcial do sistema genital (0,04%), feto macerado (0,01%), ovário aderido (0,04%). Casos de natimortalidade (0,04%), retenção placentária (0,01 %) e cervicite (0,6%) e taxas de abortos (0,88%) também foram registrados. Cocos Gram positivos (41,3%) e bastonetes Gram negativos (52,6%) foram os microrganismos mais isolados sendo Staphylococcus aureus e Escherichia colli os principais patógenos encontrados nas infeccções uterinas. Testes in vitro mostraram que esses microrganismos apresentaram maior susceptibilidade frente ao c1oranfenicol, à gentamicina e à neomicina.