Reassessment of selective agars and filtration techniques for isolation ofCampylobacter species from faeces

Bolton, F. J.; Hutchinson, D. N.; Parker, Gene

doi:10.1007/bf01963069

Cited by 53 publications

(46 citation statements)

References 17 publications

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“…In addition, the water samples collected from two different beaches showed a similar trend of significantly higher recovery of campylobacters at 37°C than at 42°C. Similar results were reported by Bolton et al (46), where the majority of campylobacters showed better recovery and culturability at 37°C than at 42°C.…”

Section: Discussionsupporting

confidence: 90%

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Effect of Incubation Temperature on the Detection of Thermophilic Campylobacter Species from Freshwater Beaches, Nearby Wastewater Effluents, and Bird Fecal Droppings

Khan

Hill

Nowak

et al. 2013

Appl Environ Microbiol

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b This large-scale study compared incubation temperatures (37°C versus 42°C) to study the detection of thermophilic Campylobacter species, including Campylobacter jejuni, C. coli, and C. lari, in various surface water samples and bird fecal droppings around Hamilton Harbor, Lake Ontario. The putative culture isolates obtained from incubation temperatures of 37 and 42°C were confirmed by Campylobacter genus-and species-specific triplex PCR assays targeting the 16S rRNA gene and the 16S-23S rRNA gene internal transcribed spacer (ITS) region. A total of 759 water, wastewater, and bird fecal dropping samples were tested. Positive amplification reactions for the genus Campylobacter were found for 454 (60%) samples incubated at 37°C, compared to 258 (34%) samples incubated at 42°C. C. jejuni (16%) and C. lari (12%) were detected significantly more frequently at the 42°C incubation temperature than at 37°C (8% and 5%, respectively). In contrast, significantly higher rates of C. coli (14%) and other Campylobacter spp. (36%) were detected at the 37°C incubation temperature than at 42°C (8% and 7%, respectively). These results were consistent across surface water, wastewater, and bird fecal dropping samples. At times, Campylobacter spp. were recovered and detected at 37°C (3% for C. jejuni, 10% for C. coli, and 3% for C. lari) when the same samples incubated at 42°C were negative. A significantly higher rate of other Campylobacter spp. was detected only at 37°C (32%) than only at 42°C (3%). These results indicate that incubation temperature can significantly influence the culturability and detection of thermophilic and other fastidious Campylobacter spp. and that a comprehensive characterization of the Campylobacter spp. in surface water, wastewaters, or bird fecal droppings will require incubation at both 37 and 42°C.

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Section: Discussionsupporting

confidence: 90%

“…jejuni. Conversely, Bolton et al (46) showed that C. fetus subsp. fetus grew better at 37°C than at 42°C.…”

Section: Discussionmentioning

confidence: 97%

Effect of Incubation Temperature on the Detection of Thermophilic Campylobacter Species from Freshwater Beaches, Nearby Wastewater Effluents, and Bird Fecal Droppings

Khan

Hill

Nowak

et al. 2013

Appl Environ Microbiol

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“…Faecal samples were examined for Campylobacter species by culture on modified charcoal cefoperazone deoxycholate agar (CCDA ; Oxoid) and by the membrane filter method (Bolton et al, 1988 ;Steele & McDermott, 1984 Campylobacter hominis sp. nov.…”

Section: Methodsmentioning

confidence: 99%

Campylobacter hominis sp. nov., from the human gastrointestinal tract.

Lawson

On²,

Logan³

et al. 2001

International Journal of Systematic and Evolutionary Microbiology

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Sequences of 16S rDNA of a novel campylobacter from faeces of healthy humans were previously shown to originate from a new taxon, ' Candidatus Campylobacter hominis ', which could not be cultured. Since phylogenetic analysis suggested that anaerobic conditions might be required for growth, an isolation strategy was developed employing initial non-selective membrane filtration onto fastidious anaerobe agar. Campylobacters were then isolated from the resulting mixed microbial flora by a dilution strategy and/or by immunomagnetic separation with genus-specific polyclonal antibody. Isolates were identified by a genus and taxon-specific PCR assay, and 16S rDNA nucleotide sequence analysis was carried out. All isolates exhibited the typical Campylobacter characteristics of being non-fermentative, oxidase-positive, catalase-negative and Gram-negative. Unusually, however, they were straight rods lacking flagella. The 16S rDNA nucleotide sequence analysis, DNA and mol % GMC were consistent with a new Campylobacter species whose nearest phylogenetic neighbours were Campylobacter gracilis and Campylobacter sputorum. The unique species status of the isolates was further confirmed by taxonomic analysis of 47 phenotypic characteristics. The name Campylobacter hominis sp. nov. is proposed for the new species, the type strain of which is NCTC 13146 T (l LMG 19568 T ).

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“…Atypical Campylobacter spp. from stools are dif®cult to culture on the selective media that are used for C. jejuni and C. coli [3,4] and multiple techniques have been combined to improve the isolation rate of species other than C. jejuni and C. coli [13,15,16]. The combined use of selective and nonselective media with a ®ltration technique has been shown to improve isolation rates [9,17].…”

Section: Resultsmentioning

confidence: 99%

Development of a combined filtration-enrichment culture followed by a one-step duplex PCR technique for the rapid detection of Campylobacter jejuni and C. coli in human faecal samples

Misawa

Kawashima

Kawamoto

et al. 2002

Journal of Medical Microbiology

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A new combined ®ltration-enrichment culture followed by a PCR technique for the rapid detection of Campylobacter jejuni and C. coli in human faeces has been developed. Only bacteria that passed through the membrane could multiply in the enrichment culture; target bacteria were detected by a one-step duplex PCR technique with combinations of primers that are speci®c for different Campylobacter spp., which should allow for the detection of a mixed infection in a single patient. A Falcon 1 cell-culture insert and 24-well tissue-culture plates were used. After 2 days, both C. jejuni and C. coli were reliably detected in diluted faeces that were seeded with as few as 10 cells which corresponds to a concentration of 10 3 cfu=g. Even allowing for the dilution of faecal samples, this represents an increase in sensitivity of two-to-three orders of magnitude over the conventional ®ltration method.

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Reassessment of selective agars and filtration techniques for isolation ofCampylobacter species from faeces

Cited by 53 publications

References 17 publications

Effect of Incubation Temperature on the Detection of Thermophilic Campylobacter Species from Freshwater Beaches, Nearby Wastewater Effluents, and Bird Fecal Droppings

Effect of Incubation Temperature on the Detection of Thermophilic Campylobacter Species from Freshwater Beaches, Nearby Wastewater Effluents, and Bird Fecal Droppings

Campylobacter hominis sp. nov., from the human gastrointestinal tract.

Development of a combined filtration-enrichment culture followed by a one-step duplex PCR technique for the rapid detection of Campylobacter jejuni and C. coli in human faecal samples

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