Campylobacter helveticus sp. nov., a new thermophilic species from domestic animals: characterization, and cloning of a species-specific DNA probe

Stanley, John R.; Burnens, André P.; Linton, Dennis; On, Stephen L. W.; Costas, M.; Owen, Robert J.

doi:10.1099/00221287-138-11-2293

Cited by 71 publications

(40 citation statements)

References 31 publications

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“…The primers and the probe of this assay were constructed to detect at least the four thermophilic campylobacters C. jejuni, C. coli, C. lari, and C. upsaliensis, which are the Campylobacter species mainly found in poultry (4,53). The primers and probe also detected C. helveticus and C. hyointestinalis, which are found mostly in cats and dogs (50) and pigs (41), respectively, and are thus of neglible importance in poultry.…”

Section: Discussionmentioning

confidence: 99%

Detection of Campylobacter spp. in ChickenFecal Samples by Real-TimePCR

Lund¹,

Nordentoft²,

Pedersen³

et al. 2004

J Clin Microbiol

180

119

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A real-time PCR assay for detecting thermophilic Campylobacter spp. directly in chicken feces has been developed. DNA was isolated from fecal material by using magnetic beads followed by PCR with a prealiquoted PCR mixture, which had been stored at ؊18°C. Campylobacter could be detected in less than 4 h, with a detection limit of 100 to 150 CFU/ml, in a fecal suspension. A bacterial internal control was added before DNA extraction to control both DNA isolation and the presence of PCR inhibitors in the samples. The assay was performed on 111 swab samples from a Danish surveillance program and compared to conventional culturing using selective enrichment. There was no statistically significant difference in performance between real-time PCR and culture by selective enrichment, and the diagnostic specificity was 0.96 with an agreement of 0.92. Therefore, the assay should be useful for screening poultry flocks for the presence of Campylobacter.Thermophilic Campylobacter spp. are the major cause of bacterial enteric infection in humans in Denmark, and the incidence of infection has been increasing in all industrialized countries, particularly since 1990 (1). The reason for this dramatic rise in the number of human cases of campylobacteriosis is not known, but poultry is often implicated as a main source of human infections, due to the high prevalence of Campylobacter in broilers. Approximately 42% of all Danish broiler flocks harbored Campylobacter at the point of slaughter in 2002 (2).As food safety has become an increasing concern for consumers, there is a growing need for fast and sensitive methods for specific detection and identification of zoonotic microorganisms. The PCR technique has several advantages over classical bacteriology with respect to detection limit, speed, and the potential for automation and has succesfully been applied to the detection of Campylobacter spp. (27,28,32,37,52).However, the conventional PCR technique has certain drawbacks. In most analyses, the PCR products are separated on an agarose gel and vizualised on a UV board, with the size of the vizualised bands as the sole confirmation of specificity (12,28). Further specificity can be added by performing hybridization using blotting of PCR products (13, 51) or a PCR enzymelinked immunosorbent assay (ELISA) procedure (9, 15), but both are laborious and time-consuming processes.A disadvantage in the diagnostic application of conventional PCR is the production of false-positive results. Theese are attributable to contamination by nucleic acids, particularly from previously amplified material (carryover) (25). Any contaminant, even the smallest airborne remnant carried over from the previous PCR procedure, may be multiplied and produce a false-positive result. The real-time mode of amplification has abolished the need to open the PCR tubes following amplification and thereby has drastically reduced the risk of carryover contamination. In addition, the liquid hybridization assay (e.g., TaqMan probes) adds further specificity to the system, c...

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Section: Discussionmentioning

confidence: 99%

Detection of Campylobacter spp. in ChickenFecal Samples by Real-TimePCR

Lund¹,

Nordentoft²,

Pedersen³

et al. 2004

J Clin Microbiol

180

119

View full text Add to dashboard Cite

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“…§Terminal fragment sizes that could not be separated in the T-RFLP analysis. Urogenital and anal bacteria of the tammar wallaby was originally isolated from cat and dog faeces (Stanley et al, 1992), is not classified as a pathogen (Mandrell et al, 2005). C. helveticus is closely related to Campylobacter upsaliensis, which, although being associated with human diseases such as abscess, gastroenteritis and bacteraemia, has an ill-defined mode of pathogenicity (Moser et al, 2001).…”

Section: Discussionmentioning

confidence: 99%

The microbiome of the cloacal openings of the urogenital and anal tracts of the tammar wallaby, Macropus eugenii

et al. 2008

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The bacterial diversity of the openings of the urogenital and anal tracts of the adult female tammar wallaby, Macropus eugenii, was determined in order to ascertain whether the physical proximity of the openings of these tracts within the cloaca affected the two populations of bacteria. Terminal restriction fragment length polymorphism (T-RFLP) analyses of 42 wallabies identified 81 different terminal fragments, indicative of diverse and complex microbiomes at these anatomical locations. Subsequent amplified rDNA restriction analysis (ARDRA) identified 72 phylotypes from the urogenital tract and 50 from the anal tract. Twenty-two of these phylotypes were common to both tracts. Phylogenetic analysis of sequenced 16S rDNA showed that 83 % of the phylotypes were unidentified species based on the premise that any sequence possessing ,97 % homology to a known bacterial species or phylotype was novel. Thus, despite the close proximity of the openings of the urogenital and anal tracts within the cloaca, the two sites retained a diverse range of distinct bacteria, with only a small percentage of overlapping species. INTRODUCTIONMetatherian (marsupial) mammals offer a unique biological opportunity to investigate the relationship between the microflora of the urogenital and anal tracts. In contrast to eutherian (placental) mammals, which possess separate openings of the digestive and urogenital systems, most marsupials possess a common posterior chamber for these systems -the cloaca (Tyndale-Briscoe, 2005; Fig. 1). This structural feature might imply that bacteria from one tract have easy access to the other. To date, the bacteria associated with these two sites have not been studied in marsupials, although in humans it has been noted that the bacteria of the urogenital tract may vary and can be influenced by bacteria growing in the anal tract and thus occasionally can cause urogenital disease (Reid et al., 2001;Tannock, 1999).The tammar wallaby, Macropus eugenii, is regarded as a model marsupial (Tyndale-Briscoe, 2005) with more than 30 years of fundamental research into its basic biology. Despite this, little is known about the bacterial populations of this animal, all published studies having used culture-based methods that are known to be biased in favour of a small percentage of the total microbiome (Hume, 1999; Lentle et al., 2006;Old & Deane, 1998;Yadav et al., 1972).In this study we used molecular-based methods targeting the 16S rDNA gene to document the bacteria at the openings of the urogenital and anal tracts of 42 female tammar wallabies and undertook a comprehensive comparison of the major phylotypes and their diversity. This is believed to be the first extensive report of the bacterial species within the openings of these two tracts in any marsupial. The results showed that despite the close anatomical proximity of the openings of these tracts, each opening possessed a diverse and complex bacterial population comprising mainly unidentified species.Abbreviations: ARDRA, amplified rDNA restriction analysis; T-...

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“…C. concisus, C. curvus, C. gracilis, C. rectus and C. showae are detected in association with the oral cavity [82]. Alternatively, C. mucosalis, C. helveticus and C. sputorum biovar faecalis are isolated from animals [114,161].…”

Section: Atypical Campylobactersmentioning

confidence: 99%

Campylobacter

et al. 2005

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-Species within the genus, Campylobacter, have emerged over the last three decades as significant clinical pathogens, particularly of human public health concern, where the majority of acute bacterial enteritis in the Western world is due to these organisms. Of particular concern are the species, C. jejuni and C. coli, which are responsible for most of these gastrointestinal-related infections. Although these organisms have already emerged as causative agents of zoonoses, several aspects of their epidemiology and pathophysiology are only beginning to emerge. Trends in increasing antibiotic resistance are beginning to emerge with oral antibiotics, which may be the drug of choice for when it is necessary to intervene chemotherapeutically. This review wishes to examine (i) emerging clinical aspects of the disease, such as Guillain Barré syndrome (GBS), (ii) the association between these organisms and poultry as a natural host, (iii) environmental aspects of Campylobacter epidemiology, (iv) the emergence of atypical campylobacters (v) emerging trends in antibiotic resistance, (vi) adoption of modern methods for the detection of campylobacters.

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Campylobacter helveticus sp. nov., a new thermophilic species from domestic animals: characterization, and cloning of a species-specific DNA probe

Cited by 71 publications

References 31 publications

Detection of Campylobacter spp. in ChickenFecal Samples by Real-TimePCR

Detection of Campylobacter spp. in ChickenFecal Samples by Real-TimePCR

The microbiome of the cloacal openings of the urogenital and anal tracts of the tammar wallaby, Macropus eugenii

Campylobacter

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