Campylobacter Bacteria Research Articles

Protein–protein interaction of HP137 with histidine kinase HP244 does not contribute to flagellar regulation in Helicobacter pylori

Flagellar motility is essential for the ability of Helicobacter pylori to colonize the gastric mucosa. Expression of the flagella is controlled by a complex regulatory cascade involving the two-component system FlgR-HP244, the sigma factors sigma54 and sigma28 and the anti-sigma28 factor FlgM. The protein-protein interaction map of H. pylori, which is based on a high-throughput two-hybrid screen (Rain et al., 2001. Nature 409, 211-215) indicated a protein-protein interaction between the gene product of ORF hp137 and both the histidine kinase HP244 and the flagellar hook protein HP908. We hypothesized that HP137 might be involved in a feedback regulatory mechanism controlling the activity of histidine kinase HP244. Here we demonstrate that HP137 does not participate in the regulation of flagellar gene expression, neither in H. pylori nor in the closely related bacterium Campylobacter jejuni.

The N-X-S/T consensus sequence is required but not sufficient for bacterial N-linked protein glycosylation

In the Gram-negative bacterium Campylobacter jejuni there is a pgl (protein glycosylation) locus-dependent general N-glycosylation system of proteins. One of the proteins encoded by pgl locus, PglB, a homolog of the eukaryotic oligosaccharyltransferase component Stt3p, is proposed to function as an oligosaccharyltransferase in this prokaryotic system. The sequence requirements of the acceptor polypeptide for N-glycosylation were analyzed by reverse genetics using the reconstituted glycosylation of the model protein AcrA in Escherichia coli. As in eukaryotes, the N-X-S/T sequon is an essential but not a sufficient determinant for N-linked protein glycosylation. This conclusion was supported by the analysis of a novel C. jejuni glycoprotein, HisJ. Export of the polypeptide to the periplasm was required for glycosylation. Our data support the hypothesis that eukaryotic and bacterial N-linked protein glycosylation are homologous processes.

Elimination of viruses, bacteria and protozoan oocysts by slow sand filtration

The decimal elimination capacity (DEC) of slow sand filters (SSF) for viruses, bacteria and oocysts of Cryptosporidium has been assessed from full-scale data and pilot plant and laboratory experiments. DEC for viruses calculated from experimental data with MS2-bacteriophages in the pilot plant filters was 1.5-2 log10. E. coli and thermotolerant coliforms (Coli44) were removed at full-scale and in the pilot plant with 2-3 log10. At full-scale, Campylobacter bacteria removal was 1 log10 more than removal of Coli44, which indicated that Coli44 was a conservative surrogate for these pathogenic bacteria. Laboratory experiments with sand columns showed 2-3 and >5-6 log10 removal of spiked spores of sulphite-reducing clostridia (SSRC; C. perfringens) and oocysts of Cryptosporidium respectively. Consequently, SSRC was not a good surrogate to quantify oocyst removal by SSF. Removal of indigenous SSRC by full-scale filters was less efficient than observed in the laboratory columns, probably due to continuous loading of these filter beds with spores, accumulation and retarded transport. It remains to be investigated if this also applies to oocyst removal by SSF. The results additionally showed that the schmutzdecke and accumulation of (in)organic charged compounds in the sand increased the elimination of microorganisms. Removal of the schmutzdecke reduced DEC for bacteria by +/-2 log10, but did not affect removal of phages. This clearly indicated that, besides biological activity, both straining and adsorption were important removal mechanisms in the filter bed for microorganisms larger than viruses.

Subcommittee on the taxonomy of Campylobacter and related bacteria:Minutes of the meeting, 31 July 2002, Paris, France

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Detection of campylobacter and Shigella species in food samples using an array biosensor.

Campylobacter and Shigella bacteria are common causes of food- and water-borne illness worldwide. There is a current need in food, medical, environmental, and military markets for a rapid and user-friendly method of detecting such pathogens. The array biosensor developed at the NRL encompasses these qualities. In this study, 25-min, sandwich immunoassays were developed for the detection of Campylobacter and Shigella species in both buffer and a variety of food and beverage samples. The limit of detection for Shigella dysenteriae in buffer and chicken carcass wash was 4.9 x 10(4) cfu mL(-)(1), whereas Campylobacter jejuni could be measured at concentrations as low as 9.7 x 10(2) cfu mL(-)(1). The limits of detection and dynamic range were found to vary depending on the sample matrix, but could be improved by running the sample over the waveguide surface for longer periods of time. Samples were run with no preconcentration or enrichment steps and little-to-no sample pretreatment prior to analysis.

Presence of Campylobacter inthe respiratory tract of broiler carcasses before and after commercial scalding

Campylobacter could be detected in the thoraco-abdominal cavity of broiler carcasses even if they were carefully eviscerated by hand with no evidence of intestinal rupture or leakage. If Campylobacter is present in the air sacs, which are unavoidably torn during evisceration, it could contaminate the thoraco-abdominal cavity of the eviscerated carcass. This study was done to determine if Campylobacter contamination is present in the respiratory tract of broilers prior to evisceration. Whole carcass rinses and respiratory tract washes were done on broiler carcasses collected at a commercial processing plant just before and just after scalding. Samples were cultured for presence and numbers of Campylobacter, Escherichia coli, coliforms, and total aerobic bacteria. Campylobacter isolates were subtyped by sequencing the short variable region of the flaA gene. The same subtypes of Campylobacter were detected in whole carcass rinse samples as in respiratory tract wash samples from individual broilers. Furthermore, the same numbers and subtypes of Campylobacter were recovered from respiratory tracts of carcasses collected before scalding and those collected after scalding. However, respiratory tracts of carcasses after scalding had higher numbers of E. coli, coliforms, and total aerobic bacteria than those tested before scalding. Although some bacterial counts were higher in the respiratory tracts of carcasses after scalding, Campylobacter counts were not. It appears that Campylobacter is present in the respiratory tracts of broilers as they enter processing, and contamination may be due to airborne bacteria during production or transport.

The genome of Campylobacter jejuni: codon and amino acid usage.

The genes from the genome of the AT-rich bacterium Campylobacter jejuni were analysed and characterised with respect to usage and amino acid usage. Codon usage is generally biased for all amino acids having synonymous codons, so that AT-rich synonyms are most frequently used. Markov chain analysis showed that codon bias and over- or underrepresentation of the corresponding tri-letter words are not related. Predicted secondary structure, lipophilicity, codon position within the gene, strand, and position on the (+)-strand were all shown to be determinants of codon usage, and these effects were in part directly explained by compositional phenomena. Codon context and the GC-content at the wobble position of the fourfold degenerate sites exert indirect effects on codon usage. The factors that affect codon usage seem to affect all amino acids, rather than selected amino acids. The usage of amino acids correlates well with the GC-content of genes, i.e. usage of amino acids encoded by GC-rich codons increases with GC-content and vice versa.

International Committee on Systematics of Prokaryotes: Subcommittee on the taxonomy of Campylobacter and related bacteria

International Committee on Systematics of Prokaryotes: Subcommittee on the taxonomy of Campylobacter and related bacteria

SAFETY OF IRISH BEEF – CONCERNS, AWARENESS AND KNOWLEDGE OF IRISH CONSUMERS

ABSTRACT A survey of Irish consumers examined their concerns for the safety of food, their level of knowledge of safe food practices and their awareness of institutional structures to ensure the safety of Irish beef. Sixty‐four percent of the sample expressed concern for the safety of food. Concerns cited include freshness of food, BSE/CJD, antibiotic residue, hygiene standards and bacteria. They perceive many causes of food poisoning and their level of awareness of pathogens varies, depending on the pathogen. The level of knowledge of safe food practice in cooking was good. However, it was lower for food storage practices. Males and those with lower levels of education were less likely to be aware of adequate food safety procedures and need to be the focus of educational programs on safe food handling. As less than half the sample (49%) was aware of institutional structures and 32% of schemes that ensure the safety of Irish beef, it is concluded that it is imperative to more clearly communicate to the public the assurances of any scheme set up. There is also a need to increase awareness level of the bacteria Campylobacter.

Microbiological Consequences of Skin Removal Prior to Evisceration of Broiler Carcasses

The objective of this project was to determine if removal of skin prior to evisceration lowers the number of bacteria that can be recovered by whole carcass rinse or sponge sampling. Four experiments were conducted, two with each type of sampling (rinse or sponge). New York dressed carcasses obtained from a commercial broiler processing plant were aseptically skinned or left with skin intact. The carcasses were then aseptically eviscerated by hand. Carcasses were rinsed in 100 mL sterile water or sampled by moist sponge. When sampled by rinse, significantly fewer Campylobacter and total aerobic bacteria were recovered from carcasses that had been skinned prior to evisceration. When sampled by sponge, significantly fewer Campylobacter, Escherichia coli, coliform and total aerobic bacteria were recovered from the outer surface of carcasses without skin. No differences were noted for bacterial counts recovered from internal surfaces by sponge sampling. Similar trends were observed when carcasses were subjected to an inside and outside washing step after evisceration. Removal of skin and washing the carcass led to significantly less Campylobacter being recovered by whole carcass rinse compared to carcasses that were washed with the skin on. When sampled by sponge, incidence of Campylobacter and level of total aerobic bacterial counts were lower on the outer surface of skinned and washed carcasses than on washed carcasses with intact skin. Like the unwashed carcasses, no differences were noted for bacterial counts recovered from internal surfaces by sponge sampling. Although not commercially practical, it is possible to lower the level of Campylobacter on the outside of broiler carcasses by removal of the skin prior to evisceration.

Trichuris suis: Detection of Antibacterial Activity in Excretory-Secretory Products from Adults

Abner, S. R., Parthasarathy, G., Hill, D. E., and Mansfield, L. S. 2001. Trichuris suis: Detection of antibacterial activity in excretory-secretory products from adults. Experimental Parasitology99, 26–36. Antibacterial activity was detected in excretory-secretory products (ESP) of adult Trichuris suis cultured in vitro in serum-free media. Gram-negative bacteria (Campylobacter jejuni, Campylobacter coli, and Escherichia coli) and Gram-positive bacteria (Staphylococcus aureus) were sensitive to ESP. Susceptibility was dependent on the concentration of ESP but not on the inoculum size. Preliminary assessment of the mode of action suggests a bacteriocidal mechanism. This antibacterial activity was heat stable and resistant to digestion with pronase E and trypsin. Based on ultrafiltration experiments, the activity is less than 10,000 MW. This excreted/secreted antibacterial activity from T. suis is likely a component of a humoral defense system for this helminth.

Taxonomy of Campylobacter, Arcobacter, Helicobacter and related bacteria: current status, future prospects and immediate concerns.

The taxonomy of the genus Campylobacter has changed dramatically since its inception in 1963. At that time the genus comprised just two species. At present, taxa that were once assigned to Campylobacter may belong to one of over 50 species distributed among six genera. Most of these taxa belong to a phylogenetically distinct group referred to as either ribosomal RNA (rRNA) superfamily VI or the epsilon division of the class Proteobacteria. The taxonomic diversity of the group is matched by the diverse habitats in which they may be found, and by the wide range of diseases that they are associated with. Recognition of their clinical and economic importance has resulted in intense interest in the group, and the application of increasingly sophisticated isolation, detection and chemotaxonomic methods continues to elucidate new aspects of their biodiversity. However, despite the advances in new bacterial systematics, there remain a number of important issues concerning the classification of various campylobacterial taxa that require careful consideration. Ultimately, these issues are relevant to many working in the field of applied microbiology, including clinicians, veterinarians, epidemiologists and taxonomists. The purpose of this article is briefly to review the major developments in the taxonomy of Campylobacter from its inception to the present day; summarize the most recent changes in the field; analyse current topical issues of special relevance to applied microbiologists, including identification of the bacteria; and speculate on future prospects for campylobacterial taxonomy.

Presence and Level of Campylobacter, Coliforms, Escherichia coli, and Total Aerobic Bacteria Recovered from Broiler Parts with and without Skin

This study was undertaken to determine if broiler chicken parts without skin are less contaminated with Campylobacter than those with skin. Samples were taken in a commercial plant from defeathered carcasses before evisceration. Bacterial counts from rinse of aseptically removed meat samples were lower than those from stomached skin samples. No Campylobacter were recovered from meat collected from the breasts or thighs, and only 2 of 10 drumstick meat samples had detectable levels of Campylobacter. However, 9 of 10 breast skin, 10 of 10 thigh skin, and 8 of 10 drumstick skin samples were positive for Campylobacter, with between 2 and 3 log10 CFU/g of Campylobacter. Breasts, thighs, and drumsticks were removed from broiler carcasses following evisceration before entering the chill tank. There was a significant difference (50 to 90%) in the levels of Campylobacter on breasts, thighs, and drumsticks with and without skin. Similar trends were noted for coliform, Escherichia coli, and total aerobic bacterial counts from samples collected in the plant. Broiler part samples were also collected at retail outlets. These samples were either skin on and skinned in the laboratory or skin off at purchase. Aseptic removal of skin from broiler breasts, thighs, and drumsticks did not cause change in Campylobacter, coliform, E. coli, or total aerobic counts recovered from the skinned part. Likewise, parts purchased without skin did not have different bacterial counts than paired parts purchased with the skin on. Consumers should not expect to significantly lower the number of bacteria present on a chicken breast, thigh, or drumstick by removing the skin.

Frequent molecular identification ofCampylobacter but notHelicobacter genus in bile and biliary epithelium in hepatolithiasis

Bacterial infection of the biliary tree and bile stasis may be causally related to hepatolithiasis, but which bacterial species are involved and their roles in the pathogenesis of hepatolithiasis have not been ascertained. Recently, the Helicobacter genus was detected in human bile and biliary mucosal samples by molecular techniques, and its association with several biliary diseases has been suggested. The Campylobacter genus, which is closely related to the Helicobacter genus, has also recently been identified as causative of human gastrointestinal diseases. This study attempted to elucidate whether Helicobacter and/or Campylobacter bacteria are present in bile samples and biliary mucosal specimens from hepatolithiasis patients and whether they are involved in the pathogenesis of hepatolithiasis. The 16S rRNA gene of the Helicobacter and of the Campylobacter genus was examined by polymerase chain reaction in DNA samples extracted from bile and/or microdissected biliary epithelium from 69 patients with hepatolithiasis and control patients with choledocholithiasis, cholecystolithiasis, and normal gall bladders. The Helicobacter genus was detected in 1 of 8 (13%) biliary epithelial samples in hepatolithiasis and 1 of 10 (10%) bile samples in choledocholithiasis. The Campylobacter genus was detected in 3 of 14 (21%) bile samples and 5 of 8 (63%) epithelial samples in hepatolithiasis, and in 2 of 15 (13%) bile samples and 1 of 8 (13%) epithelial samples in cholecystolithiasis. The detection rate for Campylobacter in biliary epithelium of hepatolithiasis was significantly higher than in the bile or biliary epithelium of control groups (p<0.05). By a phylogenetic analysis based on nucleotide sequences, the Campylobacter genuses detected in hepatolithiasis were clustered with C. rectus or C. showae. The frequent detection of the Campylobacter 16S rRNA gene in bile, and especially in biliary epithelium of hepatolithiasis, suggests a pathogenetic relationship with Campylobacter infection.

Effects of hot water application after defeathering on the levels of Campylobacter, coliform bacteria, and Escherichia coli on broiler carcasses

Scalding has been found to lower the levels of Campylobacter on broiler carcasses. However, the numbers recovered from whole-carcass rinse samples increase following defeathering. This study was undertaken to examine the effect of a second scald applied after defeathering on microbial levels recovered from carcass rinses. Four treatments were evaluated: 1) immersion at 60 C for 28 s 30 min after defeathering, 2) immersion at 60 C for 28 s immediately after defeathering, 3) spray at 73 C for 20 s 30 min after defeathering, and 4) spray at 71 C for 20 s immediately after defeathering. As reported earlier, a significant increase in Campylobacter counts per mL whole carcass rinse was noted after carcasses were defeathered. However, when applied 30 min after defeathering, neither the immersion nor the spray second scald treatments lowered the Campylobacter counts. Likewise, neither treatment had any affect on Escherichia coli or coliform bacteria counts, even though total counts were slightly reduced by the treatments. When the second scald treatment immediately followed defeathering, the same trends were observed. Campylobacter counts after the second scald remained at the postpick levels, as did counts for E. coli and coliform bacteria, but total plate counts were slightly reduced. Overall, it would appear that a postscald treatment gentle enough not to alter the carcass appearance or meat quality would not effectively lower Campylobacter, E. coli, or coliform bacteria counts.

Recommended minimal standards for describing new species of the genus Helicobacter.

The International Committee of Systematic Bacteriology Subcommittee on the taxonomy of Campylobacter and related bacteria has agreed in principle on minimum requirements for the description of new species of the genus Helicobacter. These requirements include the recommendation that the description of a putative new species or subspecies be based on examination of at least five strains in order to provide a measure of phenotypic and genotypic variation. Recommendations for required phenotypic and molecular data are presented. The requirements are consistent with the polyphasic approach to bacterial systematics.

International Committee on Systematic Bacteriology Subcommittee on the taxonomy of Campylobacter and related bacteria. Minutes of the meetings, 13 and 16 September 1999, Baltimore, USA.

International Committee on Systematic Bacteriology Subcommittee on the taxonomy of Campylobacter and related bacteria. Minutes of the meetings, 13 and 16 September 1999, Baltimore, USA.

International Committee on Systematic Bacteriology Subcommittee on the taxonomy of Campylobacter and related bacteria. Minutes of the meeting, 2 September 1998, Budapest, Hungary.

International Committee on Systematic Bacteriology Subcommittee on the taxonomy of Campylobacter and related bacteria. Minutes of the meeting, 2 September 1998, Budapest, Hungary.

Flow cytometric detection of host cell apoptosis induced by bacterial infection.

We detail two methods for detection of cell death induced by infection of a human monocytic cell line with invasive Campylobacter bacteria. Staining with a natural ligand for exposed phosphatidylserine residues coupled with propidum iodide discriminated between apoptosis and necrosis. Additionally, cells infected with a bacterial strain expressing green fluorescent protein stained with dye sensitive to mitochondrial membrane potential demonstrated a direct association of bacteria with dying cells. Analyses of cells stained by these methods employing flow cytometry enumerated proportions of cell populations undergoing either apoptosis or necrosis after bacterial infection in vitro.

High-level resistance to trimethoprim in clinical isolates of Campylobacter jejuni by acquisition of foreign genes (dfr1 and dfr9) expressing drug-insensitive dihydrofolate reductases.

The pathogenic bacterium Campylobacter jejuni has been regarded as endogenously resistant to trimethoprim. The genetic basis of this resistance was characterized in two collections of clinical isolates of C. jejuni obtained from two different parts of Sweden. The majority of these isolates were found to carry foreign dfr genes coding for resistant variants of the dihydrofolate reductase enzyme, the target of trimethoprim. The resistance genes, found on the chromosome, were dfr1 and dfr9. In about 10% of the strains, the dfr1 and dfr9 genes occurred simultaneously. About 10% of the examined isolates were found to be negative for these dfr genes and showed a markedly lower trimethoprim resistance level than the other isolates. The dfr9 and dfr1 genes were located in the context of remnants of a transposon and an integron, respectively. Two different surroundings for the dfr9 gene were characterized. One was identical to the right-hand end of the transposon Tn5393, and in the other, the dfr9 gene was flanked by only a few nucleotides of a Tn5393 sequence. The insertion of the dfr9 gene into the C. jejuni chromosome could have been mediated by Tn5393. The frequent occurrence of high-level trimethoprim resistance in clinical isolates of C. jejuni could be related to the heavy exposure of food animals to antibacterial drugs, which could lead to the acquisition of foreign resistance genes in naturally transformable strains of C. jejuni.