Discrimination of Helicobacter pullorum and Campylobacter lari by analysis of whole cell fatty acid extracts

Steinbrueckner, B.

doi:10.1016/s0378-1097(98)00443-1

Cited by 11 publications

(20 citation statements)

References 9 publications

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“…Although H. pametensis and the unnamed taxa have been found in wild birds, they have not as yet been isolated from cases of human infection. Conversely, H. pullorum has been found in poultry and associated products, and its association with enteritis and hepatic disease in humans and production of a cytolethal distending toxin clearly suggest that it is a zoonotic pathogen (1,(17)(18)(19)21). H. canadensis appears highly related to H. pullorum in 16S rRNA gene sequence comparisons, and these species also share many biochemical properties (5; this study).…”

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confidence: 71%

“…Similarly, C. jejuni is extensively distributed in nature, and untreated drinking water has been found to be an important source of human infection (7). Finally, the possible natural presence of H. canadensis in common food animals must not be overlooked, since this species closely resembles H. pullorum, an organism that is naturally present in poultry (3,17) and that is difficult to differentiate from zoonotic campylobacters such as C. lari and Campylobacter coli (18). Inadequate isolation and identification methods may contribute to an underestimation of the true prevalence and significance of such emerging pathogens, as suggested previously (5,9).…”

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confidence: 99%

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Avian Reservoirs and Zoonotic Potential of theEmerging Human Pathogen Helicobactercanadensis

Waldenström

On²,

Ottvall

et al. 2003

Appl Environ Microbiol

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A polyphasic identification approach was used to investigate the taxonomic position of Campylobacter-like isolates recovered from barnacle geese (Branta leucopsis) and Canada geese (Branta candensis). Seven strains were selected from a collection of 21 isolates and analyzed by extensive phenotypic testing; four strains were characterized by 16S rRNA gene sequence analysis. The results clearly identified the bird isolates as Helicobacter canadensis, recently described as an emerging human pathogen. This is the first report of an animal reservoir for this organism and of its presence in Europe and confirms the zoonotic potential of H. canadensis.Helicobacter canadensis, a recently described species closely resembling the enterohepatic zoonotic agent Helicobacter pullorum (5), is one of many new enteropathogens isolated from humans (1, 16). The clinical importance of this bacterium is not fully established, but it has been isolated from fecal samples of patients with enteritis (5) and from a blood culture of a patient with bacteremia (19). It has been described as an emerging pathogen (5). No animal host has hitherto been identified for H. canadensis but its closest taxonomic relative, H. pullorum, is found in poultry and retail chicken products (3,17). Here, the presence of H. canadensis in wild birds is reported for the first time and the zoonotic potential of this bacterium is discussed.A study on the transmission, ecology, and epidemiology of Campylobacter spp. (in particular Campylobacter jejuni and Campylobacter lari) in cattle and wild birds on pastured meadows in Sweden has been conducted (J. Waldenström et al., unpublished data). Fieldwork was conducted at a shore meadow on southern Gotland, southeast Sweden, where a flock of approximately 1,000 barnacle geese (Branta leucopsis) settled for 4 weeks during spring migration in 2001. To obtain samples, the flock was disturbed and sterile cotton swabs were used to collect fecal material from fresh goose droppings. Care was taken not to touch the surrounding grass, and each of the 116 samples was placed in charcoal transport medium (Transwab; BioDisc, Solna, Sweden) and stored at 4 to 8°C in a refrigerator until cultivation. Samples were then plated onto Campylobacter selective blood-free medium (45.5 g of Campylobacter selective agar base LAB M/LAB 112/liter, 2 ampoules of cefoperazone-amphotericin supplement LAB M/X 112; IDG (UK) Limited, Bury, England) and incubated at 42°C in a microaerobic atmosphere (85% N 2, 10% CO 2 , 5% O 2 ) for 72 h, at which time the media were examined for bacterial growth. Presumptive identification as Campylobacter spp. was done by limited phenotypic characterization (cell morphology and oxidase, catalase, and hippurate hydrolysis).For detailed genetic identification, the 23 Campylobacterlike isolates obtained were characterized by amplified fragment length polymorphism (AFLP)-based profiling. Chromosomal DNA was purified from bacterial cultures with the Puregene DNA isolation kit (Gentra Systems, Minneapolis, Minn.) according to...

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confidence: 99%

Avian Reservoirs and Zoonotic Potential of theEmerging Human Pathogen Helicobactercanadensis

Waldenström

On²,

Ottvall

et al. 2003

Appl Environ Microbiol

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“…This is important, since these pathogens possess few phenotypic criteria which could serve as useful markers for their unambiguous identification. For instance, both Helicobacter pullorum and Arcobacter butzleri have habitats (e.g., pigs and chicken) and disease associations (e.g., gastroenteritis) similar to those of several campylobacters, contributing to their misidentification as campylobacters by conventional phenotypic tests (1,21,36,46,52).…”

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confidence: 99%

Species-Specific Identification of Campylobacters by Partial 16S rRNA Gene Sequencing

Gorkiewicz

Feierl²,

Schober

et al. 2003

J Clin Microbiol

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Species-specific identification of campylobacters is problematic, primarily due to the absence of suitable biochemical assays and the existence of atypical strains. 16S rRNA gene (16S rDNA)-based identification of bacteria offers a possible alternative when phenotypic tests fail. Therefore, we evaluated the reliability of 16S rDNA sequencing for the species-specific identification of campylobacters. Sequence analyses were performed by using almost 94% of the complete 16S rRNA genes of 135 phenotypically characterized Campylobacter strains, including all known taxa of this genus. It was shown that 16S rDNA analysis enables specific identification of most Campylobacter species. The exception was a lack of discrimination among the taxa Campylobacter jejuni and C. coli and atypical C. lari strains, which shared identical or nearly identical 16S rDNA sequences. Subsequently, it was investigated whether partial 16S rDNA sequences are sufficient to determine species identity. Sequence alignments led to the identification of four 16S rDNA regions with high degrees of interspecies variation but with highly conserved sequence patterns within the respective species. A simple protocol based on the analysis of these sequence patterns was developed, which enabled the unambiguous identification of the majority of Campylobacter species. We recommend 16S rDNA sequence analysis as an effective, rapid procedure for the specific identification of campylobacters.Campylobacter species are important pathogens that cause a variety of diseases in humans and animals (26, 43). The most prominent members of these proteobacteria are the species Campylobacter coli and C. jejuni, the latter of which is considered the most common cause of acute bacterial enteritis worldwide (3). To date, the genus Campylobacter comprises 16 species, and among them several species other than C. jejuni and C. coli are becoming increasingly recognized as significant human pathogens (26). However, recovery and identification of these species require specialized preparatory procedures for specimens, such as filtration steps and selective incubation methods (e.g., the use of a hydrogen-enriched atmosphere) (26). Since most routine laboratories do not use these techniques, infections caused by these taxa are likely to be underdiagnosed (13,27). In addition, phenotypic tests have only a limited discriminatory potential for the distinctive identification of Campylobacter species. These pathogens are slowly growing, fastidious organisms and are considered biochemically unreactive. As a result, extensive identification schemes comprising up to 67 phenotypic features are used to correctly identify the entire spectrum of campylobacteria (40). Moreover, the phenotypic tests used in most routine laboratories lack standardization, although it is known that even minor parameters such as the inoculum size affect the results (39). The existence of biochemically atypical strains, which exhibit unusual phenotypic profiles, represents an additional challenge (36). In these cases no...

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“…Helicobacter pullorum, an enterohepatic Helicobacter species, has been isolated from humans (2,3,(14)(15)(16) as well as avian species (1,5,14). Some of the human clinical isolates were from patients with enteritis, while others were from clinically normal individuals (3,14,16).…”

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Helicobacter pullorum Outbreak in C57BL/6NTac and C3H/HeNTac Barrier-Maintained Mice

et al. 2010

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Helicobacter pullorum is a bacterial pathogen in humans. By using microaerobic culture techniques, H. pullorum was isolated from the feces of barrier-maintained mice and identified, on the basis of biochemical, restriction fragment length polymorphism, and 16S rRNA gene sequence analyses. This finding presents an opportunity to study H. pullorum pathogenesis in mice.Helicobacter pullorum, an enterohepatic Helicobacter species, has been isolated from humans (2, 3, 14-16) as well as avian species (1,5,14). Some of the human clinical isolates were from patients with enteritis, while others were from clinically normal individuals (3,14,16). Commercial broiler and layer chickens have exhibited vibrionic hepatitis and enteritis attributed to H. pullorum (14). Chickens experimentally infected with two human and two avian isolates of H. pullorum did not develop clinical signs, but both gross and microscopic cecal lesions were noted in experimentally infected chickens (4).During routine microbiological testing, Helicobacter spp. were detected by PCR in the feces of C57BL/6NTac and C3H/ HeNTac mice housed within one isolated barrier unit. Mice maintained in the affected barrier at Taconic, Inc., in Germantown, NY, were shipped to the Massachusetts Institute of Technology for evaluation. Fresh fecal samples from mice as well as cecum and colon specimens were aseptically collected at necropsy and cultured under microaerobic conditions, and bacterial isolates were subjected to biochemical characterization conditions as previously described (9). After 72 h of incubation, individual colonies from 15 C57BL/6NTac mice and 2 C3H/HeNTac mice were observed on Columbia blood agar and CVA plates containing cefoperazone, vancomycin, and amphotericin (BBL Campy agar; BBL, Sparks, MD). Analysis of samples with phase microscopy revealed spiral, motile bacteria. The bacteria were Gram negative and curved to spiral in shape. The bacteria were oxidase positive, catalase negative, urease negative, alkaline phosphatase hydrolysis negative, indoxyl acetate hydrolysis negative, and gamma-glutamyl transpeptidase negative. The isolates grew at 37°C and 42°C, but not at 25°C. Isolates were resistant to cephalothin (30 g/disc) but sensitive to nalidixic acid (30 g/disc). These biochemical and antimicrobial characteristics are identical to those of H. pullorum human isolate MIT 98-5489.The sequences of the 16S rRNA genes of two isolates (MIT 09-6633 and 09-6634) cultured from mice feces were analyzed using previously published techniques (8). Phylogenetic trees were constructed by the neighbor-joining method (13). The total RNA sequences of the two isolates (accession numbers MIT 09-6634 and MIT 09-6635) were analyzed. Comparison of the 16S rRNA sequence with those of other bacteria in our database indicated that the H. pullorum isolate was most closely related to the H. pullorum isolate (NCTC 12826) of human origin. The next most closely related was an H. pullorum chicken isolate (NCTC 12824). The percent similarity values are indicated in the phyl...

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Discrimination of Helicobacter pullorum and Campylobacter lari by analysis of whole cell fatty acid extracts

Cited by 11 publications

References 9 publications

Avian Reservoirs and Zoonotic Potential of theEmerging Human Pathogen Helicobactercanadensis

Avian Reservoirs and Zoonotic Potential of theEmerging Human Pathogen Helicobactercanadensis

Species-Specific Identification of Campylobacters by Partial 16S rRNA Gene Sequencing

Helicobacter pullorum Outbreak in C57BL/6NTac and C3H/HeNTac Barrier-Maintained Mice

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