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1.
Clin Infect Dis ; 58(11): 1579-86, 2014 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-24550377

RESUMO

Campylobacter fetus can cause intestinal illness and, occasionally, severe systemic infections. Infections mainly affect persons at higher risk, including elderly and immunocompromised individuals and those with occupational exposure to infected animals. Outbreaks are infrequent but have provided insight into sources. Source attribution of sporadic cases through case-control interviews has not been reported. The reservoirs for C. fetus are mainly cattle and sheep. Products from these animals are suspected as sources for human infections. Campylobacter fetus is rarely isolated from food, albeit selective isolation methods used in food microbiology are not suited for its detection. We hypothesize that the general population is regularly exposed to C. fetus through foods of animal origin, cross-contaminated foodstuffs, and perhaps other, as yet unidentified, routes. Campylobacter fetus infection should be suspected particularly in patients with nonspecific febrile illness who are immunocompromised or who may have been occupationally exposed to ruminants.


Assuntos
Infecções por Campylobacter/epidemiologia , Infecções por Campylobacter/patologia , Campylobacter fetus/isolamento & purificação , Doenças Transmitidas por Alimentos/epidemiologia , Doenças Transmitidas por Alimentos/patologia , Animais , Infecções por Campylobacter/microbiologia , Bovinos , Doenças dos Bovinos/microbiologia , Doenças dos Bovinos/transmissão , Doenças Transmitidas por Alimentos/microbiologia , Humanos , Hospedeiro Imunocomprometido , Exposição Ocupacional , Ovinos , Doenças dos Ovinos/microbiologia , Doenças dos Ovinos/transmissão , Zoonoses/epidemiologia , Zoonoses/transmissão
2.
Infect Immun ; 80(7): 2361-70, 2012 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-22508861

RESUMO

Campylobacter jejuni is a major cause of bacterial food-borne enteritis worldwide, and invasion into intestinal epithelial cells is an important virulence mechanism. Recently we reported the identification of hyperinvasive C. jejuni strains and created a number of transposon mutants of one of these strains, some of which exhibited reduced invasion into INT-407 and Caco-2 cells. In one such mutant the transposon had inserted into a homologue of cj1136, which encodes a putative galactosyltransferase according to the annotation of the C. jejuni NCTC11168 genome. In the current study, we investigated the role of cj1136 in C. jejuni virulence, lipooligosaccharide (LOS) biosynthesis, and host colonization by targeted mutagenesis and complementation of the mutation. The cj1136 mutant showed a significant reduction in invasion into human intestinal epithelial cells compared to the wild-type strain 01/51. Invasion levels were partially restored on complementing the mutation. The inactivation of cj1136 resulted in the production of truncated LOS, while biosynthesis of a full-length LOS molecule was restored in the complemented strain. The cj1136 mutant showed an increase in sensitivity to the bile salts sodium taurocholate and sodium deoxycholate and significantly increased sensitivity to polymyxin B compared to the parental strain. Importantly, the ability of the mutant to colonize 1-day-old chicks was also significantly impaired. This study confirms that a putative galactosyltransferase encoded by cj1136 is involved in LOS biosynthesis and is important for C. jejuni virulence, as disruption of this gene and the resultant truncation of LOS affect both colonization in vivo and invasiveness in vitro.


Assuntos
Campylobacter jejuni/enzimologia , Campylobacter jejuni/patogenicidade , Galactosiltransferases/metabolismo , Lipopolissacarídeos/biossíntese , Fatores de Virulência/metabolismo , Animais , Campylobacter jejuni/genética , Campylobacter jejuni/crescimento & desenvolvimento , Linhagem Celular , Galinhas/microbiologia , Elementos de DNA Transponíveis , Modelos Animais de Doenças , Células Epiteliais/microbiologia , Galactosiltransferases/genética , Deleção de Genes , Teste de Complementação Genética , Humanos , Mutagênese Insercional , Fatores de Virulência/genética
3.
Appl Environ Microbiol ; 77(16): 5722-9, 2011 Aug 15.
Artigo em Inglês | MEDLINE | ID: mdl-21705532

RESUMO

Many of the poultry flocks produced in the United Kingdom are colonized with Campylobacter, and the intensive nature of poultry processing usually results in contaminated carcasses. In this study, a previously reported molecular oligonucleotide probe method was used to track a specific flock-colonizing strain(s) on broiler carcasses during processing in two United Kingdom commercial poultry processing plants. Five Campylobacter-positive flocks were sampled at four points along the processing line, postbleed, postpluck, prechill, and postchill, and two Campylobacter-negative flocks processed immediately after positive flocks were sampled prechill. flaA was sequenced from Campylobacter strains isolated from these flocks, and strain-specific probes were synthesized. Skin and cecal samples were plated onto selective agar to give individual colonies, which were transferred onto membranes. These were then hybridized with the strain- and genus-specific probes. For all the 5 positive flocks, there was a significant reduction in campylobacters postbleed compared to postpluck but no subsequent fall on sampling pre- and postchill, and the strain(s) predominating on the carcasses throughout processing came from the flock being processed. This indicates that strains from the abattoir environment were not a significant cause of carcass contamination in flocks with well-established campylobacter colonization. However, negative flocks that were preceded by positive flocks were contaminated by strains that did not generally originate from the predominating strains recovered from the ceca of the previous positive flocks. This suggests that the abattoir environment has a significant role in the contamination of carcasses from negative but not fully colonized flocks.


Assuntos
Infecções por Campylobacter/veterinária , Campylobacter/isolamento & purificação , Galinhas/microbiologia , Flagelina/genética , Contaminação de Alimentos/análise , Manipulação de Alimentos , Matadouros , Análise de Variância , Animais , Campylobacter/genética , Campylobacter/crescimento & desenvolvimento , Infecções por Campylobacter/microbiologia , Contagem de Colônia Microbiana/métodos , Sondas de DNA/genética , Doenças das Aves Domésticas/microbiologia , Reino Unido
4.
Microbes Infect ; 13(4): 383-93, 2011 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-21256241

RESUMO

Mouse models have been extensively used to investigate the mechanisms of salmonellosis. However, the role of the hosts' local intestinal responses during early stages of infection remain unclear. In this study, transcript array analysis was employed to investigate regulation of gene expression in the murine intestine following oral challenge with Salmonella enterica serovar Enteritidis. Salmonella resistant C3H/HeN mice elicited only weak transcription responses in the ileum even in the presence of bacterial replication and systemic infection. This poor response was surprising given previously published results using in vitro models. Susceptible TLR4-deficient C3H/HeJ mice displayed a stronger response, suggesting a role for TLR4 in dampening the response to Salmonella. Responses of susceptible BALB/c mice were also unremarkable. In contrast, in vitro infection of murine rectal epithelial cells induced a strong transcription response consistent with previous in vitro studies. Although the pattern of genes expressed by the ileal tissue upon in vivo infection were similar in all three mouse lines, the genes up-regulated during in vitro infection were different, indicating that the responses seen in vitro do not mimic those seen in vivo. Taken together these data indicate that in vivo responses to Salmonella, at the level of the intestine, are tightly regulated by the host.


Assuntos
Perfilação da Expressão Gênica , Regulação da Expressão Gênica/imunologia , Imunidade Inata/genética , Salmonelose Animal/genética , Imunidade Adaptativa , Animais , Análise por Conglomerados , Modelos Animais de Doenças , Feminino , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Endogâmicos C3H , Mucosa Bucal/imunologia , Mucosa Bucal/microbiologia , Distribuição Aleatória , Salmonelose Animal/imunologia , Salmonella enteritidis/imunologia
5.
Appl Environ Microbiol ; 77(1): 98-107, 2011 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-21037294

RESUMO

Improved understanding of the ecology and epidemiology of Campylobacter in the poultry farm environment is key to developing appropriate farm-based strategies for preventing flock colonization. The sources of Campylobacter causing broiler flock colonization were investigated on one poultry farm and its environment, from which samples were obtained on three occasions during each of 15 crop cycles. The farm was adjacent to a dairy farm, with which there was a shared concreted area and secondary entrance. There was considerable variation in the Campylobacter status of flocks at the various sampling times, at median ages of 20, 26, and 35 days, with 3 of the 15 flocks remaining negative at slaughter. Campylobacters were recoverable from various locations around the farm, even while the flock was Campylobacter negative, but the degree of environmental contamination increased substantially once the flock was positive. Molecular typing showed that strains from house surroundings and the dairy farm were similar to those subsequently detected in the flock and that several strains intermittently persisted through multiple crop cycles. The longitudinal nature of the study suggested that bovine fecal Campylobacter strains, initially recovered from the dairy yard, may subsequently colonize poultry. One such strain, despite being repeatedly recovered from the dairy areas, failed to colonize the concomitant flock during later crop cycles. The possibility of host adaptation of this strain was investigated with 16-day-old chickens experimentally exposed to this strain naturally present in, or spiked into, bovine feces. Although the birds became colonized by this infection model, the strain may preferentially infect cattle. The presence of Campylobacter genotypes in the external environment of the poultry farm, prior to their detection in broiler chickens, confirms the horizontal transmission of these bacteria into the flock and highlights the risk from multispecies farms.


Assuntos
Infecções por Campylobacter/veterinária , Campylobacter/classificação , Campylobacter/isolamento & purificação , Portador Sadio/veterinária , Animais , Infecções por Campylobacter/epidemiologia , Portador Sadio/epidemiologia , Bovinos , Galinhas , Análise por Conglomerados , Microbiologia Ambiental , Estudos Longitudinais , Epidemiologia Molecular , Tipagem Molecular
6.
PLoS One ; 5(8): e12253, 2010 Aug 26.
Artigo em Inglês | MEDLINE | ID: mdl-20865039

RESUMO

Campylobacter jejuni strain M1 (laboratory designation 99/308) is a rarely documented case of direct transmission of C. jejuni from chicken to a person, resulting in enteritis. We have sequenced the genome of C. jejuni strain M1, and compared this to 12 other C. jejuni sequenced genomes currently publicly available. Compared to these, M1 is closest to strain 81116. Based on the 13 genome sequences, we have identified the C. jejuni pan-genome, as well as the core genome, the auxiliary genes, and genes unique between strains M1 and 81116. The pan-genome contains 2,427 gene families, whilst the core genome comprised 1,295 gene families, or about two-thirds of the gene content of the average of the sequenced C. jejuni genomes. Various comparison and visualization tools were applied to the 13 C. jejuni genome sequences, including a species pan- and core genome plot, a BLAST Matrix and a BLAST Atlas. Trees based on 16S rRNA sequences and on the total gene families in each genome are presented. The findings are discussed in the background of the proven virulence potential of M1.


Assuntos
Infecções por Campylobacter/microbiologia , Infecções por Campylobacter/veterinária , Campylobacter jejuni/genética , Genoma Bacteriano , Doenças das Aves Domésticas/microbiologia , Animais , Proteínas de Bactérias/genética , Infecções por Campylobacter/transmissão , Campylobacter jejuni/classificação , Campylobacter jejuni/isolamento & purificação , Galinhas , Mapeamento Cromossômico , Humanos , Dados de Sequência Molecular , Filogenia
7.
Int J Food Microbiol ; 139 Suppl 1: S3-15, 2010 May 30.
Artigo em Inglês | MEDLINE | ID: mdl-20153070

RESUMO

The burden of diseases caused by food-borne pathogens remains largely unknown. Importantly data indicating trends in food-borne infectious intestinal disease is limited to a few industrialised countries, and even fewer pathogens. It has been predicted that the importance of diarrhoeal disease, mainly due to contaminated food and water, as a cause of death will decline worldwide. Evidence for such a downward trend is limited. This prediction presumes that improvements in the production and retail of microbiologically safe food will be sustained in the developed world and, moreover, will be rolled out to those countries of the developing world increasingly producing food for a global market. In this review evidence is presented to indicate that the microbiological safety of food remains a dynamic situation heavily influenced by multiple factors along the food chain from farm to fork. Sustaining food safety standards will depend on constant vigilance maintained by monitoring and surveillance but, with the rising importance of other food-related issues, such as food security, obesity and climate change, competition for resources in the future to enable this may be fierce. In addition the pathogen populations relevant to food safety are not static. Food is an excellent vehicle by which many pathogens (bacteria, viruses/prions and parasites) can reach an appropriate colonisation site in a new host. Although food production practices change, the well-recognised food-borne pathogens, such as Salmonella spp. and Escherichia coli, seem able to evolve to exploit novel opportunities, for example fresh produce, and even generate new public health challenges, for example antimicrobial resistance. In addition, previously unknown food-borne pathogens, many of which are zoonotic, are constantly emerging. Current understanding of the trends in food-borne diseases for bacterial, viral and parasitic pathogens has been reviewed. The bacterial pathogens are exemplified by those well-recognized by policy makers; i.e. Salmonella, Campylobacter, E. coli and Listeria monocytogenes. Antimicrobial resistance in several bacterial food-borne pathogens (Salmonella, Campylobacter, Shigella and Vibrio spp., methicillin resistant Staphylcoccus aureas, E. coli and Enterococci) has been discussed as a separate topic because of its relative importance to policy issues. Awareness and surveillance of viral food-borne pathogens is generally poor but emphasis is placed on Norovirus, Hepatitis A, rotaviruses and newly emerging viruses such as SARS. Many food-borne parasitic pathogens are known (for example Ascaris, Cryptosporidia and Trichinella) but few of these are effectively monitored in foods, livestock and wildlife and their epidemiology through the food-chain is poorly understood. The lessons learned and future challenges in each topic are debated. It is clear that one overall challenge is the generation and maintenance of constructive dialogue and collaboration between public health, veterinary and food safety experts, bringing together multidisciplinary skills and multi-pathogen expertise. Such collaboration is essential to monitor changing trends in the well-recognised diseases and detect emerging pathogens. It will also be necessary understand the multiple interactions these pathogens have with their environments during transmission along the food chain in order to develop effective prevention and control strategies.


Assuntos
Microbiologia de Alimentos , Parasitologia de Alimentos , Doenças Transmitidas por Alimentos , Qualidade de Produtos para o Consumidor , Doenças Transmitidas por Alimentos/microbiologia , Doenças Transmitidas por Alimentos/parasitologia , Doenças Transmitidas por Alimentos/virologia , Humanos
8.
Foodborne Pathog Dis ; 7(5): 523-35, 2010 May.
Artigo em Inglês | MEDLINE | ID: mdl-20039795

RESUMO

The combination of virulence gene and antimicrobial resistance gene typing using DNA arrays is a recently developed genomics-based approach to bacterial molecular epidemiology. We have now applied this technology to 523 Salmonella enterica subsp. enterica strains collected from various host sources and public health and veterinary institutes across nine European countries. The strain set included the five predominant Salmonella serovars isolated in Europe (Enteritidis, Typhimurium, Infantis, Virchow, and Hadar). Initially, these strains were screened for 10 potential virulence factors (avrA, ssaQ, mgtC, siiD, sopB, gipA, sodC1, sopE1, spvC, and bcfC) by polymerase chain reaction. The results indicated that only 14 profiles comprising these genes (virulotypes) were observed throughout Europe. Moreover, most of these virulotypes were restricted to only one (n = 9) or two (n = 4) serovars. The data also indicated that the virulotype did not vary significantly with host source or geographical location. Subsequently, a representative subset of 77 strains was investigated using a microarray designed to detect 102 virulence and 49 resistance determinants. The results confirmed and extended the previous observations using the virulo-polymerase chain reaction screen. Strains belonging to the same serovar grouped together, indicating that the broader virulence-associated gene complement corresponded with the serovar. There were, however, some differences in the virulence gene profiles between strains belonging to an individual serovar. This variation occurred primarily within those virulence genes that were prophage encoded, in fimbrial clusters or in the virulence plasmid. It seems likely that such changes enable Salmonella to adapt to different environmental conditions, which might be reflected in serovar-specific ecology. In this strain subset a number of resistance genes were detected and were serovar restricted to a varying degree. Once again the profiles of those genes encoding resistance were similar or the same for each serovar in all hosts and countries investigated.


Assuntos
Farmacorresistência Bacteriana/genética , Infecções por Salmonella/microbiologia , Salmonella enterica/efeitos dos fármacos , Salmonella enterica/genética , Fatores de Virulência/genética , Animais , Animais Domésticos/microbiologia , Tipagem de Bacteriófagos , Europa (Continente) , Proteínas de Fímbrias/genética , Microbiologia de Alimentos , Ilhas Genômicas/genética , Genótipo , Humanos , Testes de Sensibilidade Microbiana , Análise de Sequência com Séries de Oligonucleotídeos , Reação em Cadeia da Polimerase , Prófagos/genética , Salmonelose Animal/microbiologia , Salmonella enterica/isolamento & purificação , Salmonella enterica/patogenicidade , Sorotipagem , Especificidade da Espécie
9.
Microbiology (Reading) ; 156(Pt 4): 1134-1143, 2010 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-20035004

RESUMO

Transposon mutagenesis has been applied to a hyper-invasive clinical isolate of Campylobacter jejuni, 01/51. A random transposon mutant library was screened in an in vitro assay of invasion and 26 mutants with a significant reduction in invasion were identified. Given that the invasion potential of C. jejuni is relatively poor compared to other enteric pathogens, the use of a hyper-invasive strain was advantageous as it greatly facilitated the identification of mutants with reduced invasion. The location of the transposon insertion in 23 of these mutants has been determined; all but three of the insertions are in genes also present in the genome-sequenced strain NCTC 11168. Eight of the mutants contain transposon insertions in one region of the genome (approximately 14 kb), which when compared with the genome of NCTC 11168 overlaps with one of the previously reported plasticity regions and is likely to be involved in genomic variation between strains. Further characterization of one of the mutants within this region has identified a gene that might be involved in adhesion to host cells.


Assuntos
Proteínas de Bactérias/genética , Infecções por Campylobacter/microbiologia , Campylobacter jejuni/genética , Campylobacter jejuni/patogenicidade , Proteínas de Bactérias/metabolismo , Sequência de Bases , Campylobacter jejuni/metabolismo , Elementos de DNA Transponíveis , Humanos , Dados de Sequência Molecular , Mutagênese Insercional , Mutação , Virulência
10.
Crit Rev Microbiol ; 35(1): 1-22, 2009.
Artigo em Inglês | MEDLINE | ID: mdl-19514906

RESUMO

Acquired immunity is an important factor in the epidemiology of campylobacteriosis in the developing world, apparently limiting symptomatic infection to children of less than two years. However, also in developed countries the highest incidence is observed in children under five years and the majority of Campylobacter infections are asymptomatic, which may be related to the effects of immunity and/or the ingested doses. Not accounting for immunity in epidemiological studies may lead to biased results due to the misclassification of Campylobacter-exposed but apparently healthy persons as unexposed. In risk assessment studies, health risks may be overestimated when immunity is neglected.


Assuntos
Infecções por Campylobacter/epidemiologia , Infecções por Campylobacter/prevenção & controle , Campylobacter/imunologia , Fatores Etários , Pesquisa Biomédica/métodos , Países em Desenvolvimento , Projetos de Pesquisa Epidemiológica , Humanos , Incidência , Lactente , Recém-Nascido , Medição de Risco
11.
J Neurosci Res ; 86(15): 3359-74, 2008 Nov 15.
Artigo em Inglês | MEDLINE | ID: mdl-18627035

RESUMO

An infecting strain VLA2/18 of Campylobacter jejuni was obtained from an individual with campylobacteriosis and used to prepare chicken sera by experimental infection to investigate the role of serum anti-ganglioside antibodies in Guillain-Barré syndrome. Both sera of the patient and chicken contained anti-ganglioside antibodies and anti-Lipid A (anti-Kdo2-Lipid A) antibodies directed against the lipid A portion of the bacterial lipooligosaccharide. The anti-Kdo2-Lipid A activities inhibited voltage-gated Na (Nav) channel of NSC-34 cells in culture. We hypothesized that anti-Kdo2-Lipid A antibody acts on the functional inhibition of Nav1.4. To test this possibility, a rabbit peptide antibody (anti-Nav1.4 pAb) against a 19-mer peptide (KELKDNHILNHVGLTDGPR) on the alpha subunit of Nav1.4 was produced. Anti-Nav1.4 pAb was cross-reactive to Kdo2-Lipid A. Anti-Kdo2-lipid A antibody activity in the chicken serum was tested for the Na(+) current inhibition in NSC-34 cells in combination with mu-Conotoxin and tetrodotoxin. Contrary to our expectations, the anti-Kdo2-Lipid A antibody activity was extended to Nav channels other than Nav1.4. By overlapping structural analysis, it was found that there might be multiple peptide epitopes containing certain dipeptides showing a structural similarity with v-Lipid A. Thus, our study suggests the possibility that there are multiple epitopic peptides on the extracellular domains of Nav1.1 to 1.9, and some of them may represent target sites for anti-Kdo2-Lipid A antibody, to induce neurophysiological changes in GBS by disrupting the normal function of the Nav channels.


Assuntos
Autoanticorpos/imunologia , Autoantígenos/imunologia , Infecções por Campylobacter/imunologia , Campylobacter jejuni/imunologia , Lipídeo A/imunologia , Canais de Sódio/imunologia , Sequência de Aminoácidos , Animais , Especificidade de Anticorpos , Western Blotting , Galinhas , Reações Cruzadas , Ensaio de Imunoadsorção Enzimática , Epitopos de Linfócito B/imunologia , Gangliosídeos/imunologia , Humanos , Imuno-Histoquímica , Camundongos , Dados de Sequência Molecular , Técnicas de Patch-Clamp , Peptídeos/imunologia , Isoformas de Proteínas/imunologia , Canais de Sódio/química
12.
J Med Microbiol ; 57(Pt 5): 570-580, 2008 May.
Artigo em Inglês | MEDLINE | ID: mdl-18436589

RESUMO

Campylobacter jejuni causes gastroenteritis with a variety of symptoms in humans. In the absence of a suitable animal model, in vitro models have been used to study virulence traits such as invasion and toxin production. In this study, 113 C. jejuni isolates from poultry and poultry-related (n=74) environments as well as isolates from human cases (n=39) of campylobacteriosis and bacteraemia were tested for invasiveness using INT 407 cells. The method was sufficiently reproducible to observe a spectrum of invasiveness amongst strains. As a result, strains were classified as low, high and hyper-invasive. The majority of strains (poultry and human) were low invaders (82 % and 88 %, respectively). High invasion was found for 5 % of human strains and 11 % of poultry-related isolates. However, only 1 % of poultry strains were classified as hyperinvasive compared to 13 % of human isolates (P=0.0182). Of those isolates derived from the blood of bacteraemic patients, 20 % were hyperinvasive, though this correlation was not statistically significant. An attempt was made to correlate invasiveness with the presence of seven genes previously reported to be associated with virulence. Most of these genes did not correlate with invasiveness, but gene cj0486 was weakly over-represented, and a negative correlation was observed for the gene ciaB. This trend was stronger when the two genes were analysed together, thus ciaB(-) cj0486(+) was over-represented in high and hyperinvasive strains, with low invaders more commonly found to lack these genes (P=0.0064).


Assuntos
Infecções por Campylobacter/microbiologia , Campylobacter jejuni/classificação , Galinhas/microbiologia , Doenças das Aves Domésticas/microbiologia , Animais , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Campylobacter jejuni/genética , Campylobacter jejuni/patogenicidade , Linhagem Celular Tumoral , Cloaca/microbiologia , Regulação Bacteriana da Expressão Gênica/fisiologia , Genes Bacterianos , Abrigo para Animais , Humanos , Virulência
13.
Mol Microbiol ; 68(2): 474-91, 2008 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-18284594

RESUMO

Campylobacter jejuni is a gastrointestinal pathogen of humans but can asymptomatically colonize the avian gut. C. jejuni therefore grows at both 37 degrees C and 42 degrees C, the internal temperatures of humans and birds respectively. Microarray and proteomic studies on temperature regulation in C. jejuni strain 81-176 revealed the upregulation at 42 degrees C of two proteins, Cj0414 and Cj0415, orthologous to gluconate dehydrogenase (GADH) from Pectobacterium cypripedii. 81-176 demonstrated GADH activity, converting d-gluconate to 2-keto-d-gluconate, that was higher at 42 degrees C than at 37 degrees C. In contrast, cj0414 and cj0415 mutants lacked GADH activity. Wild-type but not cj0415 mutant bacteria exhibited gluconate-dependent respiration. Neither strain grew in defined media with d-gluconate or 2-keto-d-gluconate as a sole carbon source, revealing that gluconate was used as an electron donor rather than as a carbon source. When administered to chicks individually or in competition with wild-type, the cj0415 mutant was impaired in establishing colonization. In contrast, there were few significant differences in colonization of BALB/c-ByJ mice in single or mixed infections. These results suggest that the ability of C. jejuni to use gluconate as an electron donor via GADH activity is an important metabolic characteristic that is required for full colonization of avian but not mammalian hosts.


Assuntos
Proteínas de Bactérias/metabolismo , Campylobacter jejuni/enzimologia , Gluconatos/metabolismo , Oxirredutases/metabolismo , Animais , Proteínas de Bactérias/genética , Infecções por Campylobacter/microbiologia , Campylobacter jejuni/química , Campylobacter jejuni/crescimento & desenvolvimento , Campylobacter jejuni/metabolismo , Ceco/microbiologia , Galinhas , Contagem de Colônia Microbiana , Eletroforese em Gel Bidimensional , Deleção de Genes , Perfilação da Expressão Gênica , Camundongos , Camundongos Endogâmicos BALB C , Análise de Sequência com Séries de Oligonucleotídeos , Oxirredutases/genética , Oxigênio/metabolismo , Pectobacterium/enzimologia , Pectobacterium/genética , Proteoma/análise , Homologia de Sequência de Aminoácidos , Temperatura , Fatores de Virulência/genética , Fatores de Virulência/metabolismo
14.
Appl Environ Microbiol ; 74(8): 2321-31, 2008 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-18281428

RESUMO

The identification of sites resulting in cross-contamination of poultry flocks in the abattoir and determination of the survival and persistence of campylobacters at these sites are essential for the development of intervention strategies aimed at reducing the microbial burden on poultry at retail. A novel molecule-based method, using strain- and genus-specific oligonucleotide probes, was developed to detect and enumerate specific campylobacter strains in mixed populations. Strain-specific oligonucleotide probes were designed for the short variable regions (SVR) of the flaA gene in individual Campylobacter jejuni strains. A 16S rRNA Campylobacter genus-specific probe was also used. Both types of probes were used to investigate populations of campylobacters by colony lift hybridization. The specificity and proof of principle of the method were tested using strains with closely related SVR sequences and mixtures of these strains. Colony lifts of campylobacters were hybridized sequentially with up to two labeled strain-specific probes, followed by the generic 16S rRNA probe. SVR probes were highly specific, differentiating down to 1 nucleotide in the target sequence, and were sufficiently sensitive to detect colonies of a single strain in a mixed population. The 16S rRNA probe detected all Campylobacter spp. tested but not closely related species, such as Arcobacter skirrowi and Helicobacter pullorum. Preliminary field studies demonstrated the application of this technique to target strains isolated from poultry transport crate wash tank water. This method is quantitative, sensitive, and highly specific and allows the identification and enumeration of selected strains among all of the campylobacters in environmental samples.


Assuntos
Campylobacter jejuni/isolamento & purificação , Contagem de Colônia Microbiana/métodos , Hibridização de Ácido Nucleico/métodos , Sequência de Bases , Campylobacter jejuni/genética , Microbiologia Ambiental , Flagelina/genética , Microbiologia de Alimentos , Dados de Sequência Molecular , Sondas de Oligonucleotídeos/genética , RNA Bacteriano/genética , RNA Ribossômico 16S/genética , Sensibilidade e Especificidade , Alinhamento de Sequência
15.
Appl Environ Microbiol ; 74(8): 2492-504, 2008 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-18203857

RESUMO

Reducing colonization of poultry flocks by Campylobacter spp. is a key strategy in the control and prevention human campylobacteriosis. Horizontal transmission of campylobacters, from in and around the farm, is the presumed route of flock colonization. However, the identification and prioritization of sources are confounded by the ubiquitous nature of these organisms in the environment, their poor rates of recovery by standard culture methods, and the need for cost-effective and timely methods for strain-specific comparison. A real-time PCR screening test for the strain-specific detection of campylobacters in environmental samples has been developed to address this issue. To enable this approach, fluorescently labeled PCR oligonucleotide probes suitable for a LightCycler-based assay were designed to match a highly variable DNA segment within the flaA short variable region (SVR) of Campylobacter jejuni or C. coli. The capacity of such probes to provide strain-specific tools was investigated by using bacterial cultures and spiked and naturally contaminated poultry fecal and environmental samples. The sensitivity of two representative probes was estimated, by using two different C. jejuni strains, to be 1.3 x 10(2) to 3.7 x 10(2) CFU/ml in bacterial cultures and 6.6 x 10(2) CFU/ml in spiked fecal samples. The specificity of the SVR for C. jejuni and C. coli was confirmed by using a panel of strains comprising other Campylobacter species and naturally contaminated samples. The approach was field tested by sampling the environment and feces of chickens of two adjacently located poultry houses on a conventional broiler farm throughout the life of one flock. All environmental samples were enriched for 2 days, and then DNA was prepared and stored. Where feasible, campylobacter isolates were also recovered and stored for subsequent testing. A strain-specific probe based on the SVR of the strain isolated from the first positive chicken fecal sample was developed. This probe was then used to screen the stored environmental samples by real-time PCR. Pulsed-field gel electrophoresis was used to compare recovered environmental and fecal isolates to assess the specificity of the method. The results established the proof of principle that strain-specific probes, based on the SVR of flaA, can identify a flock-colonizing strain in DNA preparations from enriched environmental cultures. Such a novel strategy provides the opportunity to investigate the epidemiology of campylobacters in poultry flocks and allows targeted biosecurity interventions to be developed. The strategy may also have wider applications for the tracking of specific campylobacter strains in heavily contaminated environments.


Assuntos
Infecções por Campylobacter/microbiologia , Campylobacter coli/isolamento & purificação , Campylobacter jejuni/isolamento & purificação , Microbiologia Ambiental , Reação em Cadeia da Polimerase/métodos , Doenças das Aves Domésticas/microbiologia , Animais , Sequência de Bases , Campylobacter coli/genética , Campylobacter jejuni/genética , Galinhas , Análise por Conglomerados , Impressões Digitais de DNA , Primers do DNA/genética , DNA Bacteriano/genética , Eletroforese em Gel de Campo Pulsado , Fezes/microbiologia , Flagelina/genética , Genótipo , Epidemiologia Molecular , Dados de Sequência Molecular , Sensibilidade e Especificidade , Análise de Sequência de DNA , Homologia de Sequência , Temperatura de Transição
16.
Microb Pathog ; 43(5-6): 198-207, 2007.
Artigo em Inglês | MEDLINE | ID: mdl-17600669

RESUMO

The contribution of gamma-glutamyl transpeptidase (GGT) to Campylobacter jejuni virulence and colonization of the avian gut has been investigated. The presence of the ggt gene in C. jejuni strains directly correlated with the expression of GGT activity as measured by cleavage and transfer of the gamma-glutamyl moiety. Inactivation of the monocistronic ggt gene in C. jejuni strain 81116 resulted in isogenic mutants with undetectable GGT activity; nevertheless, these mutants grew normally in vitro. However, the mutants had increased motility, a 5.4-fold higher invasion efficiency into INT407 cells in vitro and increased resistance to hydrogen peroxide stress. Moreover, the apoptosis-inducing activity of the ggt mutant was significantly lower than that of the parental strain. In vivo studies showed that, although GGT activity was not required for initial colonization of 1-day-old chicks, the enzyme was required for persistent colonization of the avian gut.


Assuntos
Infecções por Campylobacter/veterinária , Campylobacter jejuni/fisiologia , Trato Gastrointestinal/microbiologia , gama-Glutamiltransferase/fisiologia , Adaptação Biológica/imunologia , Animais , Aves , Infecções por Campylobacter/microbiologia , Campylobacter jejuni/enzimologia , Campylobacter jejuni/genética , Campylobacter jejuni/patogenicidade , Galinhas/microbiologia , gama-Glutamiltransferase/genética
17.
Microbiology (Reading) ; 153(Pt 5): 1339-1349, 2007 May.
Artigo em Inglês | MEDLINE | ID: mdl-17464048

RESUMO

Despite being classically defined as non-pathogenic, there is growing evidence that biotype 1A Yersinia enterocolitica isolates may be aetiological agents of disease in humans. In previous studies, a potential link between motility and the ability of biotype 1A strains to invade cultured epithelial cells was observed. In an attempt to further investigate this finding, a flagella mutant was constructed in a human faecal Y. enterocolitica biotype 1A isolate. The flagella mutation abolished the ability of the strain to invade cultured human epithelial cells, although adherence was not affected. The aflagellate mutant was also attenuated in its ability to survive within cultured macrophages, being cleared after 3 h, whilst the wild-type persisted for 24 h after infection. Examination of cytokine secretion by infected macrophages also suggested that the flagella of biotype 1A strains act as anti-inflammatory agents, decreasing production of tumour necrosis factor (TNF)-alpha whilst increasing secretion of interleukin (IL)-10. Preliminary studies using porcine in vitro organ culture (IVOC) tissue suggested that the flagella mutant was also attenuated in its ability to colonize intestinal tissue.


Assuntos
Citocinas/biossíntese , Células Epiteliais/microbiologia , Flagelos/fisiologia , Macrófagos/microbiologia , Virulência/genética , Yersiniose/microbiologia , Yersinia enterocolitica/crescimento & desenvolvimento , Animais , Aderência Bacteriana , Linhagem Celular , Colo/microbiologia , Contagem de Colônia Microbiana , Fezes/microbiologia , Flagelos/genética , Humanos , Íleo/microbiologia , Microscopia Eletrônica de Transmissão , Técnicas de Cultura de Órgãos , Suínos , Yersinia enterocolitica/genética , Yersinia enterocolitica/imunologia , Yersinia enterocolitica/isolamento & purificação
18.
J Med Microbiol ; 55(Pt 12): 1725-1734, 2006 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-17108278

RESUMO

Previous epidemiological studies have demonstrated a potential link between the serotypes of Yersinia enterocolitica recovered from cattle, sheep and pigs and those isolated from human disease cases. Further studies utilizing amplified fragment length polymorphisms have shown a relationship at the genetic level between strains of biotypes 3 and 4 from humans and livestock, and also suggested that some biotype 1A isolates, classically defined as non-pathogenic, are closely related to biotype 3 and 4 isolates. This study sought to understand further the pathogenic potential of Y. enterocolitica isolates from livestock in Great Britain. A range of surrogate in vitro models, such as invasion of epithelial tissue cultures, survival in cultured macrophages and cytokine secretion response, was employed to assess the pathogenicity of 88 strains. The results suggested that all isolates examined were capable of adhering to and invading epithelial cells and of surviving within macrophages. However, the inflammatory response of the infected macrophages differed with the infecting Y. enterocolitica subtype, with the response to pathogenic biotype 3 and 4 isolates different to that observed with biotype 1A isolates, and with the biotype 3 O : 5,27 isolates recovered exclusively from animals. Infections of porcine tissue also suggested the possibility of host-tissue tropism within Y. enterocolitica subtypes.


Assuntos
Citocinas/biossíntese , Yersiniose/microbiologia , Yersinia enterocolitica/fisiologia , Animais , Aderência Bacteriana , Bovinos , Linhagem Celular , Células Epiteliais/imunologia , Células Epiteliais/microbiologia , Humanos , Macrófagos/imunologia , Macrófagos/microbiologia , Ovinos , Especificidade da Espécie , Suínos , Reino Unido , Yersinia enterocolitica/crescimento & desenvolvimento , Yersinia enterocolitica/imunologia
19.
Avian Dis ; 50(1): 1-9, 2006 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-16617973

RESUMO

Epidemiological studies indicate that Campylobacter species may be responsible for the majority of cases of sporadic gastroenteritis in humans. These studies also suggest that poultry may be one of the most common sources of the bacteria for humans. Campylobacter and related genera in the family Campylobacteraceae are oral and intestinal commensals of vertebrates and some nonvertebrates, a characteristic that complicates rational approaches to controlling Campylobacter contamination of poultry. This review will discuss the phylogeny, genomics, and physiology of campylobacters with the intention of revealing how these organisms have evolved to fill their intestinal ecological niche in poultry and how their physiology must be understood in order to enact effective control strategies.


Assuntos
Infecções por Campylobacter/veterinária , Doenças das Aves Domésticas/microbiologia , Animais , Campylobacter/genética , Campylobacter/isolamento & purificação , Campylobacter/fisiologia , Infecções por Campylobacter/epidemiologia , Infecções por Campylobacter/microbiologia , Infecções por Campylobacter/prevenção & controle , Galinhas/microbiologia , Microbiologia de Alimentos , Humanos , Carne/microbiologia , Doenças das Aves Domésticas/epidemiologia , Doenças das Aves Domésticas/prevenção & controle
20.
Exp Neurol ; 200(1): 50-5, 2006 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-16500643

RESUMO

Campylobacteriosis is frequently associated with Guillain-Barré syndrome. Poultry are frequently highly colonized with Campylobacter jejuni and are a major foodborne vehicle for campylobacteriosis. In this study, high titer anti-GM1 antibodies were found in the serum of a laboratory worker who developed campylobacteriosis. The microbiologically confirmed strain VLA2/18 (non-serotyped) was isolated from the worker and subsequently inoculated into chickens, resulting in high titers of serum antibodies to GM1. However, none of the immunized chickens in our study showed any noticeable neurological symptoms, such as paralysis or cramping. High titer anti-GM1 antibodies in chicken and human sera strongly inhibited spontaneous muscle action potential in an in vitro system of spinal cord and muscle cell co-culture. In addition, infection of chickens with C. jejuni strains 81116 (HS6) and 99/419 (HS21) or immunization with purified GM1, GM2, and GM3 resulted in elevation of serum anti-ganglioside antibodies with an inhibitory effect on spontaneous muscle action potential. Immunoabsorption studies demonstrated that this inhibitory activity is due to anti-ganglioside antibodies. On the other hand, anti-GM1 is the only specific human serum antibody to induce an inhibitory effect on neuromuscular junctions. Chicken anti-GM1 antibodies showed a strong inhibitory effect, but anti-GM2 and -GM3 had weaker activities. Taken together, our data suggest that campylobacteriosis in chickens may provide a strong link between infection and the development of anti-ganglioside antibody-mediated peripheral nerve dysfunctions.


Assuntos
Anticorpos Antibacterianos/sangue , Infecções por Campylobacter/imunologia , Campylobacter jejuni/imunologia , Gangliosídeos/imunologia , Doenças das Aves Domésticas/imunologia , Potenciais de Ação/imunologia , Animais , Anticorpos Antibacterianos/biossíntese , Infecções por Campylobacter/prevenção & controle , Campylobacter jejuni/isolamento & purificação , Galinhas , Técnicas de Cocultura , Humanos , Doenças das Aves Domésticas/prevenção & controle , Ratos , Especificidade da Espécie
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