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1.
PLoS One ; 10(10): e0139636, 2015.
Artigo em Inglês | MEDLINE | ID: mdl-26431341

RESUMO

BACKGROUND: Campylobacteriosis is the most frequently reported food- and waterborne infection in Norway. We investigated the risk factors for sporadic Campylobacter infections in Norway in order to identify areas where control and prevention measures could be improved. METHODS: A national prospective case-control study of factors associated with Campylobacter infection was conducted from July 2010 to September 2011. Cases were recruited from the Norwegian Surveillance System of Communicable Diseases (MSIS). Controls were randomly selected from the Norwegian Population Registry. Cases and controls were mailed a paper questionnaire with a prepaid return envelope. Univariable analyses using logistic regression were conducted for all exposures. A final parsimonious multivariable model was developed using regularized/penalized logistic regression, and adjusted odds ratios were calculated. RESULTS: A total of 995 cases and 1501 controls were included in the study (response proportion 55% and 30%, respectively). Exposures that had significant increases in odds of Campylobacter infection in multivariable analysis were drinking water directly from river, stream, or lake (OR: 2.96), drinking purchased bottled water (OR: 1.78), eating chicken (1.69), eating meat that was undercooked (OR: 1.77), eating food made on a barbecue (OR: 1.55), living on a farm with livestock (OR: 1.74), having a dog in the household (OR: 1.39), and having household water supply serving fewer than 20 houses (OR: 1.92). CONCLUSIONS: Consumption of poultry and untreated water remain important sources of Campylobacter infection in Norway, despite ongoing control efforts. The results justify the need for strengthening education for consumers and food handlers about the risks of cross-contamination when preparing poultry and with consuming raw or undercooked chicken. The public should also be reminded to take precautions when drinking untreated water in nature and ensure continued vigilance in order to protect and maintain the quality of water from small-scale water supply systems.


Assuntos
Infecções por Campylobacter/epidemiologia , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Estudos de Casos e Controles , Criança , Pré-Escolar , Feminino , Humanos , Lactente , Recém-Nascido , Masculino , Pessoa de Meia-Idade , Noruega , Estudos Prospectivos , Fatores de Risco , Adulto Jovem
2.
BMC Infect Dis ; 15: 324, 2015 Aug 11.
Artigo em Inglês | MEDLINE | ID: mdl-26259588

RESUMO

BACKGROUND: Shiga toxin-producing E. coli (STEC) infection is associated with haemolytic uremic syndrome (HUS). Therefore Norway has implemented strict guidelines for prevention and control of STEC infection. However, only a subgroup of STEC leads to HUS. Thus, identification of determinants differentiating high risk STEC (HUS STEC) from low risk STEC (non-HUS STEC) is needed to enable implementation of graded infectious disease response. METHODS: A national study of 333 STEC infections in Norway, including one STEC from each patient or outbreak over two decades (1992-2012), was conducted. Serotype, virulence profile, and genotype of each STEC were determined by phenotypic or PCR based methods. The association between microbiological properties and demographic and clinical data was assessed by univariable analyses and multiple logistic regression models. RESULTS: From 1992 through 2012, an increased number of STEC cases including more domestically acquired infections were notified in Norway. O157 was the most frequent serogroup (33.6 %), although a decrease of this serogroup was seen over the last decade. All 25 HUS patients yielded STEC with stx2, eae, and ehxA. In a multiple logistic regression model, age ≤5 years (OR = 16.7) and stx2a (OR = 30.1) were independently related to increased risk of HUS. eae and hospitalization could not be modelled since all HUS patients showed these traits. The combination of low age (≤5 years) and the presence of stx2a, and eae gave a positive predictive value (PPV) for HUS of 67.5 % and a negative predictive value (NPV) of 99.0 %. SF O157:[H7] and O145:H?, although associated with HUS in the univariable analyses, were not independent risk factors. stx1 (OR = 0.1) was the sole factor independently associated with a reduced risk of HUS (NPV: 79.7 %); stx2c was not so. CONCLUSIONS: Our results indicate that virulence gene profile and patients' age are the major determinants of HUS development.


Assuntos
Infecções por Escherichia coli/epidemiologia , Síndrome Hemolítico-Urêmica/epidemiologia , Escherichia coli Shiga Toxigênica/isolamento & purificação , Adolescente , Adulto , Criança , Pré-Escolar , Surtos de Doenças , Infecções por Escherichia coli/microbiologia , Infecções por Escherichia coli/prevenção & controle , Proteínas de Escherichia coli/genética , Feminino , Genótipo , Síndrome Hemolítico-Urêmica/microbiologia , Síndrome Hemolítico-Urêmica/prevenção & controle , Humanos , Lactente , Masculino , Pessoa de Meia-Idade , Noruega/epidemiologia , Reação em Cadeia da Polimerase/métodos , Fatores de Risco , Sorogrupo , Virulência/genética , Adulto Jovem
3.
Appl Environ Microbiol ; 78(12): 4083-91, 2012 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-22492457

RESUMO

A previous national survey of Escherichia coli in Norwegian sheep detected eae-positive (eae(+)) E. coli O26:H11 isolates in 16.3% (80/491) of the flocks. The purpose of the present study was to evaluate the human-pathogenic potential of these ovine isolates by comparing them with E. coli O26 isolates from humans infected in Norway. All human E. coli O26 isolates studied carried the eae gene and shared flagellar type H11. Two-thirds of the sheep flocks and 95.1% of the patients harbored isolates containing arcA allele type 2 and espK and were classified as enterohemorrhagic E. coli (EHEC) (stx positive) or EHEC-like (stx negative). These isolates were further divided into group A (EspK2 positive), associated with stx(2-EDL933) and stcE(O103), and group B (EspK1 positive), associated with stx(1a). Although the stx genes were more frequently present in isolates from patients (46.3%) than in those from sheep flocks (5%), more than half of the ovine isolates in the EHEC/EHEC-like group had multiple-locus variable number of tandem repeat analysis (MLVA) profiles that were identical to those seen in stx-positive human O26:H11 isolates. This indicates that EHEC-like ovine isolates may be able to acquire stx-carrying bacteriophages and thereby have the possibility to cause serious illness in humans. The remaining one-third of the sheep flocks and two of the patients had isolates fulfilling the criteria for atypical enteropathogenic E. coli (aEPEC): arcA allele type 1 and espK negative (group C). The majority of these ovine isolates showed MLVA profiles not previously seen in E. coli O26:H11 isolates from humans. However, according to their virulence gene profile, the aEPEC ovine isolates should be considered potentially pathogenic for humans. In conclusion, sheep are an important reservoir of human-pathogenic E. coli O26:H11 isolates in Norway.


Assuntos
Reservatórios de Doenças , Escherichia coli/isolamento & purificação , Escherichia coli/patogenicidade , Ovinos/microbiologia , Animais , Análise por Conglomerados , Escherichia coli/genética , Proteínas de Escherichia coli/genética , Humanos , Tipagem Molecular , Noruega , Fatores de Virulência/genética
4.
FEMS Microbiol Lett ; 329(2): 102-10, 2012 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-22268961

RESUMO

Sorbitol-fermenting Escherichia coli O157:NM (SF O157) is an emerging pathogen suggested to be more virulent than nonsorbitol-fermenting Escherichia coli O157:H7 (NSF O157). Important virulence factors are the Shiga toxins (stx), encoded by stx1 and/or stx2 located within prophages integrated in the bacterial genome. The stx genes are expressed from p(R) (') as a late protein, and anti-terminator activity from the Q protein is necessary for read through of the late terminator t(R) (') and activation of p(R) (') . We investigated the regulation of stx2(EDL933) expression at the genomic level in 17 Norwegian SF O157. Sequencing of three selected SF O157 strains revealed that the anti-terminator q gene and genes upstream of stx2(EDL933) were identical or similar to the ones observed in the E. coli O111:H- strain AP010960, but different from the ones observed in the NSF O157 strain EDL933 (AE005174). This suggested divergent stx2(EDL933) -encoding bacteriophages between NSF O157 and the SF O157 strains (FR874039-41). Furthermore, different DNA structures were detected in the SF O157 strains, suggesting diversity among bacteriophages also within the SF O157 group. Further investigations are needed to elucidate whether the q(O111:H) (-) gene observed in all our SF O157 contributes to the increased virulence seen in SF O157 compared to NSF O157. An assay for detecting q(O111:H) (-) was developed.


Assuntos
Escherichia coli O157/genética , Proteínas de Escherichia coli/genética , Regiões Promotoras Genéticas/genética , Toxina Shiga II/genética , Sorbitol/metabolismo , Sequência de Bases , Infecções por Escherichia coli/microbiologia , Escherichia coli O157/classificação , Escherichia coli O157/metabolismo , Escherichia coli O157/patogenicidade , Proteínas de Escherichia coli/metabolismo , Fermentação , Regulação Bacteriana da Expressão Gênica , Humanos , Dados de Sequência Molecular , Prófagos/genética , Toxina Shiga II/metabolismo
5.
BMC Infect Dis ; 8: 41, 2008 Apr 03.
Artigo em Inglês | MEDLINE | ID: mdl-18387178

RESUMO

BACKGROUND: On 20-21 February 2006, six cases of diarrhoea-associated haemolytic uraemic syndrome (HUS) were reported by paediatricians to the Norwegian Institute of Public Health. We initiated an investigation to identify the etiologic agent and determine the source of the outbreak in order to implement control measures. METHODS: A case was defined as a child with diarrhoea-associated HUS or any person with an infection with the outbreak strain of E. coli O103 (defined by the multi-locus variable number tandem repeats analysis (MLVA) profile) both with illness onset after January 1st 2006 in Norway. After initial hypotheses-generating interviews, we performed a case-control study with the first fifteen cases and three controls for each case matched by age, sex and municipality. Suspected food items were sampled, and any E. coli O103 strains were typed by MLVA. RESULTS: Between 20 February and 6 April 2006, 17 cases were identified, of which 10 children developed HUS, including one fatal case. After pilot interviews, a matched case-control study was performed indicating an association between a traditional cured sausage (odds ratio 19.4 (95% CI: 2.4-156)) and STEC infection. E. coli O103:H25 identical to the outbreak strain defined by MLVA profile was found in the product and traced back to contaminated mutton. CONCLUSION: We report an outbreak caused by a rare STEC variant (O103:H25, stx2-positive). More than half of the diagnosed patients developed HUS, indicating that the causative organism is particularly virulent. Small ruminants continue to be important reservoirs for human-pathogen STEC. Improved slaughtering hygiene and good manufacturing practices for cured sausage products are needed to minimise the possibility of STEC surviving through the entire sausage production process.


Assuntos
Surtos de Doenças , Infecções por Escherichia coli/epidemiologia , Síndrome Hemolítico-Urêmica/epidemiologia , Produtos da Carne/microbiologia , Escherichia coli Shiga Toxigênica/isolamento & purificação , Adolescente , Animais , Estudos de Casos e Controles , Criança , Pré-Escolar , Infecções por Escherichia coli/microbiologia , Fezes/microbiologia , Abastecimento de Alimentos , Síndrome Hemolítico-Urêmica/microbiologia , Humanos , Lactente , Entrevistas como Assunto , Repetições Minissatélites , Noruega/epidemiologia , Sorotipagem , Soro/microbiologia , Ovinos , Escherichia coli Shiga Toxigênica/classificação , Escherichia coli Shiga Toxigênica/genética
6.
Foodborne Pathog Dis ; 5(2): 165-73, 2008 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-18361685

RESUMO

On November 15, 2004, a cluster of three cases of Salmonella Thompson infection was registered by the Norwegian reference laboratory. In the following days further cases occurred, prompting a case-control study among the first 13 cases and 26 matched controls. By December 31, 21 cases had been reported, with the first onset on October 24. Consumption of rucola lettuce (Eruca sativa, also known as rocket salad or arugula) (OR 8,8 [1,2-infinity]) and mixed salad (OR 5,0 [1,0-infinity]) was associated with illness. On November 26, Swedish authorities notified the finding of Salmonella Thompson in rucola lettuce through the EU Rapid Alert System for Food and Feed. Later, several countries reported finding this and other Salmonella serovars and Campylobacter in rucola produced in Italy. In response to our alert through the international Enter-net surveillance network, Sweden and England also reported an increase of cases. Salmonella Thompson isolates from products and patients from several countries showed high similarity by pulsed-field gel electrophoresis, but some isolates showed significant differences. We think that the outbreak in Norway reflected a larger international outbreak caused by rucola imported from one Italian producer. Findings of other pathogens indicate a massive contamination, possibly caused by irrigation with nonpotable water. Rapid international information exchange is invaluable when investigating outbreaks caused by internationally marketed products.


Assuntos
Contaminação de Alimentos/análise , Lactuca/microbiologia , Intoxicação Alimentar por Salmonella/epidemiologia , Salmonella/isolamento & purificação , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Estudos de Casos e Controles , Criança , Pré-Escolar , Surtos de Doenças , Feminino , Microbiologia de Alimentos , Humanos , Lactente , Masculino , Pessoa de Meia-Idade , Noruega/epidemiologia , Vigilância da População , Salmonella/classificação
7.
J Microbiol Methods ; 72(2): 141-8, 2008 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-18096258

RESUMO

The multiple-locus variable-number tandem-repeats analysis (MLVA) method for genotyping has proven to be a fast and reliable typing tool in several bacterial species. MLVA is in our laboratory the routine typing method for Salmonella enterica subsp. enterica serovar Typhimurium and Escherichia coli O157. The gram-positive bacteria Listeria monocytogenes, while not isolated as frequent as S. Typhimurium and E. coli, causes severe illness with an overall mortality rate of 30%. Thus, it is important that any outbreak of this pathogen is detected early and a fast trace to the source can be performed. In view of this, we have used the information provided by two fully sequenced L. monocytogenes strains to develop a MLVA assay coupled with high-resolution capillary electrophoresis and compared it to pulsed-field gel electrophoresis (PFGE) in two sets of isolates, one Norwegian (79 isolates) and one Swedish (61 isolates) set. The MLVA assay could resolve all of the L. monocytogenes serotypes tested, and was slightly more discriminatory than PFGE for the Norwegian isolates (28 MLVA profiles and 24 PFGE profiles) and opposite for the Swedish isolates (42 MLVA profiles and 43 PFGE profiles).


Assuntos
Técnicas de Tipagem Bacteriana/métodos , Eletroforese Capilar/métodos , Eletroforese em Gel de Campo Pulsado/métodos , Listeria monocytogenes/classificação , Repetições Minissatélites , DNA Bacteriano/genética , Humanos , Listeria monocytogenes/genética , Listeria monocytogenes/isolamento & purificação , Dados de Sequência Molecular , Noruega , Reação em Cadeia da Polimerase , Suécia
8.
Emerg Infect Dis ; 13(5): 754-6, 2007 May.
Artigo em Inglês | MEDLINE | ID: mdl-17553258

RESUMO

An outbreak involving 11 persons infected with Yersinia enterocolitica O:9 was investigated in Norway in February 2006. A case-control study and microbiologic investigation indicated a ready-to-eat pork product as the probable source. Appropriate control measures are needed to address consumer risk associated with this product.


Assuntos
Surtos de Doenças , Manipulação de Alimentos , Produtos da Carne/microbiologia , Yersiniose/epidemiologia , Yersinia enterocolitica/patogenicidade , Adulto , Idoso , Idoso de 80 Anos ou mais , Animais , Estudos de Casos e Controles , Criança , Enterite/epidemiologia , Enterite/microbiologia , Doenças Transmitidas por Alimentos/microbiologia , Humanos , Pessoa de Meia-Idade , Noruega/epidemiologia , Suínos , Yersiniose/etiologia , Yersinia enterocolitica/classificação
9.
J Microbiol Methods ; 69(1): 197-205, 2007 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-17291612

RESUMO

The Escherichia coli (E. coli) reference collection, ECOR, consists of 72 strains that are representative of the genotypic diversity, as indexed by multilocus enzyme electrophoresis (MLEE), in the species as a whole. MLEE revealed 4 main phylogenetic groups designated A, B1, B2 and D. We present a study of the relationship between the ECOR strains as determined by polymorphisms in seven variable-number of tandem repeats (VNTR) loci. Seven tandem repeats that were present in more than one of the fully sequenced E. coli strains were selected, and primers were constructed in order to amplify the targets in all species where the loci were present. The combined result for all VNTR loci was adapted as a multiple-locus variable-number tandem repeats analysis (MLVA) and showed that the ECOR collection was divided into 63 distinct genotypes. The ECOR phylogenetic groups defined by MLEE were not well conserved by MLVA. A set of 61 pathogenic isolates of both E. coli and Shigella spp. was then tested with the same set of VNTR loci, and revealed 54 distinct genotypes. In addition, the MLVA method was used to genotype isolates from patients and suspected sources in a recent outbreak of E. coli O103 in Norway. The pathogenic E. coli isolates contained the diarrhea causing categories EIEC, EAEC, STEC, ETEC and EPEC. Shigella isolates were of species S. flexneri, S. boydii, S. sonnei and S. dysenteriae. The MLVA method rapidly genotyped all isolates in the study at a Simpson's index of diversity of D=0.98.


Assuntos
Técnicas de Tipagem Bacteriana/métodos , Escherichia coli/classificação , Repetições Minissatélites , Shigella/classificação , Escherichia coli/genética , Escherichia coli/patogenicidade , Genótipo , Noruega , Filogenia , Polimorfismo Genético , Shigella/genética , Shigella/patogenicidade
10.
J Microbiol Methods ; 68(2): 331-41, 2007 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-17079041

RESUMO

A multiplex PCR assay, amplifying seven specific virulence genes and one internal control gene in a single reaction, was developed to identify the five main pathotypes of diarrheagenic Escherichia coli and Shigella spp. The virulence genes selected for each category were Stx1, Stx2, and eaeA for enterohemorrhagic E. coli (EHEC), eaeA for enteropathogenic E. coli (EPEC), STIb and LTI for enterotoxigenic E. coli (ETEC), ipaH for enteroinvasive E. coli (EIEC) and Shigella spp., and aggR for enteroaggregative E. coli (EAEC). Each forward primer was labelled with a fluorochrome and the PCR products were separated by multicolour capillary electrophoresis on an ABI PRISM310 Genetic Analyzer (Applied Biosystems). If present, several gene variants of each virulence gene were identified. The internal control gene rrs, encoding 16S rRNA, was amplified in all 110 clinical strains analyzed. Virulence genes were demonstrated in 103 (94%) of these strains. In the majority of the cases (98/103, 95%), classification obtained by the novel multiplex PCR assay was in agreement with that previously determined by phenotypic assays combined with other molecular genetic approaches. Numerous multiplex PCR assays have been published, but only a few of them detect all five E. coli pathotypes within a single reaction, and none of them has used multicolour capillary electrophoresis to separate the PCR products. The octaplex PCR assay followed by capillary electrophoresis presented in the present paper provides a simple, reliable, and rapid procedure that in a single reaction identifies the five main pathotypes of E. coli, and Shigella spp. This assay will replace the previous molecular genetic methods used in our laboratory and work as an important supplement to the more time-consuming phenotypic assays.


Assuntos
Diarreia/microbiologia , Disenteria Bacilar/microbiologia , Eletroforese Capilar/métodos , Infecções por Escherichia coli/microbiologia , Escherichia coli/genética , Reação em Cadeia da Polimerase/métodos , Shigella/genética , Adesinas Bacterianas/química , Adesinas Bacterianas/genética , Antígenos de Bactérias/química , Antígenos de Bactérias/genética , Proteínas de Bactérias/química , Proteínas de Bactérias/genética , DNA Bacteriano/química , DNA Bacteriano/genética , Escherichia coli/isolamento & purificação , Escherichia coli/patogenicidade , Proteínas de Escherichia coli/química , Proteínas de Escherichia coli/genética , Fluorescência , Humanos , Estudos Retrospectivos , Toxina Shiga I/química , Toxina Shiga I/genética , Toxina Shiga II/química , Toxina Shiga II/genética , Shigella/isolamento & purificação , Shigella/patogenicidade , Transativadores/química , Transativadores/genética , Virulência
12.
Int J Food Microbiol ; 96(1): 85-96, 2004 Oct 01.
Artigo em Inglês | MEDLINE | ID: mdl-15358509

RESUMO

We have investigated the molecular epidemiology of Listeria monocytogenes from the meat processing industry producing cold cuts and from cases of human listeriosis by discriminative pulsed-field gel electrophoresis (PFGE). A subset of the isolates was also investigated for susceptibility to a disinfectant based on quaternary ammonium compounds (QAC) frequently used in the meat processing industry. The purpose of this investigation was to obtain knowledge of sources, routes of contamination and genetic types of L. monocytogenes present along the production line in the meat processing industry, and to compare meat industry isolates and human isolates. Of the 222 isolates from four meat-processing plants, 200 were from two plants responsible for nearly 50% of the production of cold cuts in the Norwegian market. The strain collection included historical routinely sampled isolates (1989-2002) and isolates systematically sampled through a one year period (November 2001 to November 2002) from fresh meat and production environments in three plants. No isolates were obtained in samples from employees (throat, faeces). Human strains included all available reported isolates from Norwegian patients in selected time periods. The L. monocytogenes PFGE data showed a large genetic heterogeneity, with isolates separated into two genetic lineages and further subdivided into 56 different PFGE profiles. Certain profiles were observed on both sides of production (before and after heat treatment) indicating contamination of end products by fresh meat or fresh meat environments. While fresh meat isolates almost exclusively grouped within lineage I, isolates from end products showed a more balanced distribution between lineages I and II. Ten profiles were common among isolates from human and meat industry. Typing of human isolates identified a previously unrecognised outbreak. Generally, a higher QAC resistance incidence was observed among isolates from the meat processing industry than among human isolates although large plant to plant differences were indicated. No correlation between resistance and PFGE profile or resistance and persistence was observed.


Assuntos
Desinfetantes/farmacologia , Manipulação de Alimentos/métodos , Indústria de Processamento de Alimentos/normas , Listeria monocytogenes , Produtos da Carne/microbiologia , Compostos de Amônio Quaternário/farmacologia , Animais , Farmacorresistência Bacteriana , Eletroforese em Gel de Campo Pulsado , Contaminação de Alimentos/análise , Humanos , Listeria monocytogenes/efeitos dos fármacos , Listeria monocytogenes/genética , Listeria monocytogenes/isolamento & purificação , Testes de Sensibilidade Microbiana
13.
J Microbiol Methods ; 59(2): 163-72, 2004 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-15369852

RESUMO

The multiple-locus variable-number tandem-repeats analysis (MLVA) method is currently being used as the primary typing tool for Salmonella enterica subsp. enterica serovar Typhimurium (S. Typhimurium) isolates in our laboratory. Our published initial MLVA was performed using a single fluorescent dye and the different patterns were assigned from gel images. Here we present a new and significantly improved assay using multiple dye colors, enhanced PCR multiplexing and the introduction of two new loci for better adaptation to capillary electrophoresis with increased speed. The different MLVA patterns are now based on allele sizes entered as character values, thus removing the uncertainties introduced when analyzing band patterns from gel images. We additionally propose an easy numbering scheme for the identification of separate isolates that will facilitate exchange of typing data. A total of 106 human, bird, animal and food isolates of S. Typhimurium, including 16 with definite type (DT) 104, were used for the development of the improved MLVA. The method is based on capillary separation of multiplexed PCR products from five VNTR loci in the S. Typhimurium genome labeled with multiple fluorescent dyes. The different alleles at each locus were then assigned allele numbers, which were used for strain comparison.


Assuntos
Reação em Cadeia da Polimerase/métodos , Salmonella typhimurium/classificação , Salmonella typhimurium/genética , Alelos , Animais , Impressões Digitais de DNA/métodos , DNA Bacteriano/química , DNA Bacteriano/genética , Eletroforese Capilar/métodos , Corantes Fluorescentes/química , Humanos , Repetições Minissatélites/genética , Filogenia
14.
Ann Clin Microbiol Antimicrob ; 3: 12, 2004 Jul 08.
Artigo em Inglês | MEDLINE | ID: mdl-15242512

RESUMO

BACKGROUND: Class 1 integrons contain genetic elements for site-specific recombination, capture and mobilization of resistance genes. Studies investigating the prevalence, distribution and types of integron located resistance genes are important for surveillance of antimicrobial resistance and to understand resistance development at the molecular level. METHODS: We determined the prevalence and genetic content of class 1 integrons in Enterobacteriaceae (strain collection 1, n = 192) and E. coli (strain collection 2, n = 53) from bloodstream infections in patients from six Norwegian hospitals by molecular techniques. Class 1 integrons were also characterized in 54 randomly selected multiresistant E. coli isolates from gastrointestinal human infections (strain collection 3). RESULTS: Class 1 integrons were present in 10.9% of the Enterobacteriaceae blood culture isolates of collection 1, all but one (S. Typhi) being E. coli. Data indicated variations in class 1 integron prevalence between hospitals. Class 1 integrons were present in 37% and 34% of the resistant blood culture isolates (collection 1 and 2, respectively) and in 42% of the resistant gastrointestinal E. coli. We detected a total of 10 distinct integron cassette PCR amplicons that varied in size between 0.15 kb and 2.2 kb and contained between zero and three resistance genes. Cassettes encoding resistance to trimethoprim and aminoglycosides were most common. We identified and characterized a novel plasmid-located integron with a cassette-bound novel gene (linF) located downstream of an aadA2 gene cassette. The linF gene encoded a putative 273 aa lincosamide nucleotidyltransferase resistance protein and conferred resistance to lincomycin and clindamycin. The deduced LinF amino acid sequence displayed approximately 35% identity to the Enterococcus faecium and Enterococcus faecalis nucleotidyl transferases encoded by linB and linB' CONCLUSIONS: The present study demonstrated an overall low and stable prevalence of class 1 integron gene cassettes in clinical Enterobacteriaceae and E. coli isolates in Norway. Characterization of the novel lincosamide resistance gene extends the growing list of class 1 integron gene cassettes that confer resistance to an increasing number of antibiotics.

15.
J Microbiol Methods ; 58(2): 213-22, 2004 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-15234519

RESUMO

The Multiple-Locus Variable-Number Tandem-Repeats Analysis (MLVA) method is currently being used as the primary typing tool for Shiga-toxin-producing Escherichia coli (STEC) O157 isolates in our laboratory. The initial assay was performed using a single fluorescent dye and the different patterns were assigned using a gel image. Here, we present a significantly improved assay using multiple dye colors and enhanced PCR multiplexing to increase speed, and ease the interpretation of the results. The different MLVA patterns are now based on allele sizes entered as character values, thus removing the uncertainties introduced when analyzing band patterns from the gel image. We additionally propose an easy numbering scheme for the identification of separate isolates that will facilitate exchange of typing data. Seventy-two human and animal strains of Shiga-toxin-producing E. coli O157 were used for the development of the improved MLVA assay. The method is based on capillary separation of multiplexed PCR products of VNTR loci in the E. coli O157 genome labeled with multiple fluorescent dyes. The different alleles at each locus were then assigned to allele numbers, which were used for strain comparison.


Assuntos
Técnicas Bacteriológicas , Escherichia coli O157/genética , Técnicas Genéticas , Repetições Minissatélites , Animais , Sequência de Bases , Primers do DNA/genética , DNA Bacteriano/genética , Eletroforese Capilar/métodos , Escherichia coli O157/classificação , Escherichia coli O157/isolamento & purificação , Corantes Fluorescentes , Humanos , Reação em Cadeia da Polimerase/métodos
16.
Ann Clin Microbiol Antimicrob ; 2: 12, 2003 Dec 10.
Artigo em Inglês | MEDLINE | ID: mdl-14664722

RESUMO

BACKGROUND: The ability to react early to possible outbreaks of Escherichia coli O157:H7 and to trace possible sources relies on the availability of highly discriminatory and reliable techniques. The development of methods that are fast and has the potential for complete automation is needed for this important pathogen. METHODS: In all 73 isolates of shiga-toxin producing E. coli O157 (STEC) were used in this study. The two available fully sequenced STEC genomes were scanned for tandem repeated stretches of DNA, which were evaluated as polymorphic markers for isolate identification. RESULTS: The 73 E. coli isolates displayed 47 distinct patterns and the MLVA assay was capable of high discrimination between the E. coli O157 strains. The assay was fast and all the steps can be automated. CONCLUSION: The findings demonstrate a novel high discriminatory molecular typing method for the important pathogen E. coli O157 that is fast, robust and offers many advantages compared to current methods.

17.
Am J Epidemiol ; 158(3): 234-42, 2003 Aug 01.
Artigo em Inglês | MEDLINE | ID: mdl-12882945

RESUMO

In 1999-2000, a prospective case-control study of sporadic, domestically acquired campylobacteriosis was conducted in three counties in Norway to identify preventable risk factors and potentially protective factors. A total of 212 cases and 422 population controls matched by age, sex, and geographic area were enrolled. In conditional logistic regression analysis, the following factors were found to be independently associated with an increased risk of Campylobacter infection: drinking undisinfected water, eating at barbecues, eating poultry bought raw, having occupational exposure to animals, and eating undercooked pork. The following factors were independently related to a decreased risk: eating mutton, eating raw fruits or berries, and swimming. Results indicated that infection is more likely to occur as a result of cross-contamination from raw poultry products than because of poultry consumption per se. Drinking undisinfected water, reported by 53% of cases, was a leading risk factor in this study. Drinking water may constitute the common reservoir linking infection in humans and animals, including poultry and wild birds. Insight into the ecology of Campylobacter in freshwater ecosystems may be required to understand the epidemiology of campylobacteriosis. The possibility that certain foods confer protection against campylobacteriosis deserves exploration.


Assuntos
Infecções por Campylobacter/epidemiologia , Infecções por Campylobacter/etiologia , Contaminação de Alimentos , Abastecimento de Água , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Estudos de Casos e Controles , Criança , Pré-Escolar , Feminino , Frutas , Geografia , Humanos , Lactente , Masculino , Carne , Pessoa de Meia-Idade , Noruega/epidemiologia , Estudos Prospectivos , Análise de Regressão , Fatores de Risco , Natação
18.
J Clin Microbiol ; 41(4): 1469-79, 2003 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-12682132

RESUMO

Seventy-eight human and environmental strains of Salmonella enterica subsp. enterica serovar Typhimurium, as well as 18 isolates of other Salmonella serovars and 6 isolates of Escherichia coli, were subjected to a novel variable number of tandem repeats (VNTR)-based fingerprinting method that showed high discrimination and reproducibility for typing serovar Typhimurium isolates. The method is based on capillary separation of PCR products from fluorescence-labeled VNTR in the serovar Typhimurium genome. The serovar Typhimurium isolates displayed 54 VNTR patterns, and the VNTR assay correctly identified strains from a well-characterized outbreak. Among 37 serovar Typhimurium phage type DT104 isolates, 28 distinct VNTR patterns were found. This VNTR-based method is fast and suitable for complete automation. Our VNTR-based method was capable of high discrimination within the homogeneous serovar Typhimurium DT104 phage type and can be used to trace outbreaks and to monitor DT104 as well as other phage types. The VNTR assay was compared to XbaI pulsed-field gel electrophoresis, amplified fragment length polymorphism analysis, integron-cassette profiles and gene PCR of intI1, qacEDelta1, sulI1, and floR. The VNTR assay showed greatly improved resolution compared to all other tested methods in this study.


Assuntos
Impressões Digitais de DNA/métodos , Repetições Minissatélites/genética , Salmonella typhimurium/classificação , Animais , Técnicas de Tipagem Bacteriana , Eletroforese em Gel de Campo Pulsado , Microbiologia de Alimentos , Humanos , Reação em Cadeia da Polimerase , Polimorfismo de Fragmento de Restrição , Infecções por Salmonella/microbiologia , Salmonelose Animal/microbiologia , Salmonella typhimurium/virologia , Sorotipagem
19.
J Wildl Dis ; 39(1): 64-72, 2003 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-12685069

RESUMO

Septicemic salmonellosis caused by Salmonella Typhimurium 4, 12: i:1, 2 was diagnosed in 94 (64.8%) of 145 small passerines comprising nine species, examined in Norway during 1999-2000. The birds were found dead at private feeding places throughout the country. The bullfinch (Pyrrhula pyrrhula), Eurasian siskin (Carduelis spinus), common redpoll (Carduelis flammea), and Eurasian greenfinch (Carduelis chloris) were the most frequently affected species. Pathologic findings in 94 carcasses included poor body condition (84%), enlarged spleen (73%), and necrosis of crop/esophagus (78%), liver (53%), spleen (46%), proventriculus (13%), and intestine (5.3%). Histologically, necrosis consisted of debris, fibrin, inflammatory cells, and aggregates of Gram-negative bacteria and occasionally giant cells. Based on information from questionnaires sick and dead birds were observed at feeding places from December to June, with a distinct peak during February and March. The duration of recorded outbreaks varied from less than 1 wk to 4 mo. In a separate study, 1,990 apparently healthy passerines caught at feeding places established for bird-ringing purposes were surveyed for cloacal carriage of Salmonella spp. Forty (2.0%) of the birds examined, representing sampling sites both in southern and northern parts of the country, harbored S. Typhimurium 4, 12: i:1, 2 in their intestines. The carrier species largely reflected the species most often suffering from fatal infection.


Assuntos
Doenças das Aves/mortalidade , Doenças das Aves/patologia , Salmonelose Animal/mortalidade , Salmonelose Animal/patologia , Salmonella typhimurium , Aves Canoras , Animais , Portador Sadio/epidemiologia , Portador Sadio/patologia , Portador Sadio/veterinária , Surtos de Doenças/veterinária , Feminino , Masculino , Noruega/epidemiologia , Salmonella typhimurium/isolamento & purificação , Salmonella typhimurium/patogenicidade , Aves Canoras/microbiologia , Especificidade da Espécie , Baço/microbiologia , Baço/patologia , Inquéritos e Questionários
20.
J Med Microbiol ; 52(Pt 2): 141-9, 2003 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-12543920

RESUMO

The characterization of integrons and their promoters in 156 antibiotic-resistant clinical isolates of the important zoonotic pathogens Salmonella enterica subsp. enterica serovars Typhimurium (S. Typhimurium) and Enteritidis (S. Enteritidis) from Norwegian hospitals were performed. Integrons were found in 64 of 66 S. Typhimurium isolates (97 %) and in 20 of 90 S. Enteritidis isolates (22.2 %) with the following sizes; 650, 1000, 1200, 1500, 1600, 1700, 2000 and 2100 bp. The integrons were further sequenced and the aadA1, aadA2, aadA5, aadB, pse-1, catB3, oxa1, dfrA1, dfrA12 and dfrA17 genes, as well as a fragment of the sat1 gene, were found embedded in cassettes. An internal fragment of the purG gene was additionally found as an artefact PCR amplicon.


Assuntos
Integrons , Infecções por Salmonella/microbiologia , Salmonella enteritidis/genética , Salmonella typhimurium/genética , Antibacterianos/farmacologia , Tipagem de Bacteriófagos , Sequência de Bases , DNA Bacteriano , Farmacorresistência Bacteriana/genética , Farmacorresistência Bacteriana Múltipla/genética , Humanos , Testes de Sensibilidade Microbiana , Dados de Sequência Molecular , Peso Molecular , Noruega , Plasmídeos , Reação em Cadeia da Polimerase/métodos , Regiões Promotoras Genéticas/genética , Salmonella enteritidis/efeitos dos fármacos , Salmonella typhimurium/efeitos dos fármacos , Homologia de Sequência do Ácido Nucleico
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