PubMed Health⌕ Search

Biomedical subjects

S L On

Publications and source records attributed to S L On.

At least 19 recordsLinked to original sources
PubMed2001

Serotype and genotype diversity and hatchery transmission of Campylobacter jejuni in commercial poultry flocks.

We investigated the genotype and serotype diversity of Campylobacter coli and C. jejuni in two parent flocks of adult hens and their offspring over two rotations in order to evaluate the role of hatchery mediated transmission and/or vertical transmission of campylobacters in broiler flocks. In total, 314 C. jejuni and 32 C. coli isolates from parent and broiler flocks and from the surroundings of broiler houses were typed by flagellin gene PCR/RFLP (fla-typing), and selected isolates were also typed by serotyping and macrorestriction profiling using PFGE (MRP/PFGE). The combined typing results showed that the broiler flocks could be colonised by 1-3 different Campylobacter clones and parent flocks could be colonised by 2-6 different clones. C. coli was isolated from up to 36% of birds in one parent flock, whereas only C. jejuni was isolated from broiler flocks. C. jejuni clones from different flocks were clearly discriminated by fla-typing as well as by MRP/PFGE, except for a few cases where individual isolates belonging to two different clones were found to have altered fla-types. Similarly, one C. coli clone showed pronounced fla-type variation. The present results lead to the conclusion that vertical transmission or horizontal transmission via the hatchery are not significant transmission routes of C. jejuni to broiler chickens under Danish conditions. In the cases where more than one Campylobacter clone simultaneously colonised flocks, we found that the different clones coexisted in flocks rather than excluding each other.

Animals↗
PubMed2001

Evaluation of numerical analysis of PFGE-DNA profiles for differentiating Campylobacter fetus subspecies by comparison with phenotypic, PCR and 16S rDNA sequencing methods.

AIMS: To assess the efficacy of numerical analysis of PFGE-DNA profiles for identification and differentiation of Campylobacter fetus subspecies. METHODS AND RESULTS: 31 Camp. fetus strains were examined by phenotypic, PCR- and PFGE-based methods, and the 16S rDNA sequences of 18 strains compared. Numerical analysis of PFGE-DNA profiles divided strains into two clusters at the 86% similarity level. One cluster contained 19 strains clearly identified as Camp. fetus subsp. venerealis. The other cluster comprised 12 strains, of which 10 were unambiguously identified as Camp. fetus subsp. fetus. The remaining two strains were identified as Camp. fetus subsp. venerealis by either phenotypic or PCR methods, but not both. At higher similarity levels, clusters containing isolates from each of two countries were identified, suggesting that certain clones predominate in certain geographical regions. CONCLUSION: Numerical analysis of PFGE-DNA profiles is an effective method for differentiating Camp. fetus subspecies. SIGNIFICANCE AND IMPACT OF THE STUDY: Critical comparison of PFGE, PCR, 16S rDNA sequencing and phenotypic methods for differentiation of Camp. fetus subspecies was attained. Novel phenotypic markers for distinguishing subspecies were identified. Evidence for dominant clones of each subspecies in certain countries was provided.

Animals↗
PubMed2001

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

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% G+C 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 13146T (= LMG 19568T).

Bacterial Typing Techniques↗
PubMed2001

Recommendations of the subcommittee on the taxonomy of Campylobacter and related bacteria.

The ICSB Subcommittee on the taxonomy of Campylobacter and related bacteria has discussed several contemporaneous issues and makes the following recommendations. (i) The reported synonymy between Campylobacter coli and Campylobacter hyoilei was based on valid taxonomic arguments. The subcommittee therefore discourages the use of the name C. hyoilei. (ii) The revised infrasubspecific nomenclature of Campylobacter sputorum is endorsed. C sputorum is subdivided into C. sputorum biovar sputorum (characterized by the absence of catalase and urease activity); C. sputorum biovar faecalis (characterized by catalase but not urease activity); and C. sputorum biovar paraureolyticus (characterized by urease, but not catalase activity). (iii) The subcommittee points out that 'Flexispira rappini' is a taxon that is circumscribed by means of morphological criteria. It encompasses multiple Helicobacter species, including Helicobacter billis and Helicobacter trogontum. (iv) Finally, the subcommittee wishes to point out that the etymology of several specific or subspecific epithets of Campylobacter taxa has been corrected and that the spelling of the epithet 'fecalis' was corrected to 'faecalis'.

Campylobacter↗
PubMed2001

Helicobacter ganmani sp. nov., a urease-negative anaerobe isolated from the intestines of laboratory mice.

Spiral bacteria were isolated from the intestines of laboratory mice during a study examining the presence of Helicobacter species and other spiral organisms naturally infecting mice maintained at four different animal facilities in Sydney, Australia. One group of 17 isolates, cultured from mice from three of the four facilities, were found to be helicobacters but did not fall within any of the 18 currently recognized species. These isolates were unusual in that they only grew anaerobically at 37 degrees C and were incapable of growth under microaerobic conditions. Like Helicobacter rodentium, isolates possessed single, bipolar, unsheathed flagella and were urease-negative. They were positive for oxidase and reduced nitrate to nitrite but did not hydrolyse hippurate or indoxyl acetate, grew on charcoal agar and were resistant to cephalothin. 16S rDNA sequences from four strains were determined and found to be identical to one another. H. rodentium was the most closely related species in terms of 16S rDNA sequence similarity (98.2%). Numerical analysis of whole-cell proteins by SDS-PAGE for nine isolates was carried out with a comparison to all known Helicobacter species, including newly determined profiles from three H. rodentium strains. The new isolates were clearly differentiated from H. rodentium and other Helicobacter spp. On the basis of this data, including genetic, biochemical and protein analysis, it is proposed that these isolates belong to Helicobacter ganmani sp. nov. (type strain CMRI H02T = CCUG 43526T = CIP 106846T).

Anaerobiosis↗
PubMed2001

Comparison of genotypes and serotypes of Campylobacter jejuni isolated from Danish wild mammals and birds and from broiler flocks and humans.

The incidence of human infection with Campylobacter jejuni is increasing in most developed countries and the reason for this is largely unknown. Although poultry meat is considered to be a major source, it is evident that other reservoirs exist, possibly common to humans and poultry. Environmental sources are believed to be important reservoirs of Campylobacter infection in broiler chicken flocks. We investigated the potential importance of wildlife as a source of infection in commercial poultry flocks and in humans by comparing the serotype distributions, fla types, and macrorestriction profiles (MRPs) of C. jejuni isolates from different sources. The serotype distribution in wildlife was significantly different from the known distributions in broilers and humans. Considerable sero- and genotype diversity was found within the wildlife collection, although two major groups of isolates within serotype O:12 and the O:4 complex were found. Common clonal lines among wildlife, chicken, and/or human isolates were identified within serotype O:2 and the O:4 complex. However, MRPs of O:12 and O:38 strains isolated from wildlife and other sources indicated that some clonal lines propagated in a wide selection of animal species but were not detected in humans or broilers in this study. The applied typing methods successfully identified different clonal groups within a strain collection showing large genomic diversity. However, the relatively low number of wildlife strains with an inferred clonal relationship to human and chicken strains suggests that the importance of wildlife as a reservoir of infection is limited.

Animals↗
PubMed2000

Identification of taxonomic and epidemiological relationships among Campylobacter species by numerical analysis of AFLP profiles.

Amplified fragment length polymorphism (AFLP)-based profiling was performed on 138 strains representing all named Campylobacter species and subspecies. Profiles of 15/16 species comprised 6 to greater than 100 fragments and were subjected to numerical analysis. The mean similarity of 48 duplicate, outbreak and/or 'identical' strain profiles exceeded 94%. Species were clearly distinguished at the 17.90% similarity (S-) level in the dendrogram. Subspecies of Campylobacter jejuni and Campylobacter hyointestinalis, and biovars of Campylobacter lari and Campylobacter sputorum were distinguished at higher S-levels. All outbreak or 'genetically identical' strains of C. jejuni subsp. jejuni, Campylobacter coli, C. hyointestinalis and C. sputorum clustered at S-levels >92% and were distinguished from unrelated strains. Numerical analysis of AFLP profiles is useful for concurrent identification of taxonomic and epidemiological relationships among most Campylobacter species.

Bacterial Typing Techniques↗
PubMed2000

Efficacy of flagellin gene typing for epidemiological studies of Campylobacter jejuni in poultry estimated by comparison with macrorestriction profiling.

Thirty isolates of Campylobacter jejuni isolated from 29 different Danish broiler flocks were chosen for the evaluation of PCR-Fla typing as a genotyping tool. Except for two isolates that originated from the same broiler flock, the isolates were clearly distinguishable on basis of their macrorestriction profiles using the restriction endonucleases SmaI and KpnI. PCR-Fla typing of the 30 isolates yielded 16 distinct genotypes, whereas one isolate was untypeable by this method. The dominant PCR-Fla type (1/1) was shared by eight isolates, and five additional Fla groups containing two or three isolates were obtained. The PCR-Fla type of one isolate changed spontaneously after five subcultures, illustrating the relative plasticity of the gene locus. Comparison of MRPs within and between Fla-types support the view that some PCR-Fla types may be conserved within clonal lines. It is concluded that PCR-Fla typing is useful as a genotyping tool in large-scale epidemiological studies but that additional analyses with other methods are required to properly define interstrain relationships.

Animals↗
PubMed2000

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.

Animals↗
PubMed2000

Isolation and characterization of Campylobacter jejuni subsp. jejuni from macaroni penguins (Eudyptes chrysolophus) in the subantarctic region.

On Bird Island, South Georgia, albatrosses (n = 140), penguins (n = 100), and fur seals (n = 206) were sampled for Campylobacter jejuni. C. jejuni subsp. jejuni was recovered from three macaroni penguins (Eudyptes chrysolophus). These isolates, the first reported for the subantarctic region, showed low genetic diversity and high similarity to Northern Hemisphere C. jejuni isolates, possibly suggesting recent introduction to the area.

Animals↗
PubMed2000

Prevalence of Campylobacter, Arcobacter, Helicobacter, and Sutterella spp. in human fecal samples as estimated by a reevaluation of isolation methods for Campylobacters.

The aims of this study were to investigate the prevalence of campylobacteria including Campylobacter jejuni subsp. jejuni (C. jejuni) and Campylobacter coli in human clinical samples and in samples from healthy individuals and to reevaluate the efficacies of conventional selective methods for isolation of Campylobacter spp. Two charcoal-based selective media, modified charcoal cefoperazone deoxycholate agar (mCCDA) and cefoperazone-amphotericin-teicoplanin (CAT) agar, were compared with Skirrow's blood-based medium and with a filter method (filter) applied to a yeast-enriched blood agar. A total of 1,376 specimens were tested on all four media, and the percentages of thermophilic Campylobacter-positive specimens isolated on Skirrow's medium, filters, CAT agar, and mCCDA were 82, 83, 85, and 95%, respectively. When additional samples were processed with the three selective media, mCCDA recovered significantly more thermophilic Campylobacter spp. than Skirrow's medium (P = 0.0034). No significant difference between Skirrow's medium and CAT agar was observed in this study. Another six taxa were identified, namely, Campylobacter concisus, Campylobacter curvus-like bacteria, Arcobacter butzleri, Arcobacter cryaerophilus, Helicobacter cinaedi, and Sutterella wadsworthensis. Most of these strains were isolated after 5 to 6 days of incubation by use of the filter technique. This paper provides evidence for the existence of S. wadsworthensis in human feces from clinical cases of gastrointestinal disorders and in feces from a healthy individual. Furthermore, C. concisus was isolated from a large number of diarrheal cases, particularly those at the extremes of age, but was additionally isolated from the feces of healthy people. Further investigations to establish the role of C. concisus and S. wadsworthensis in enteric disease is needed. We conclude that a range of campylobacteria may cause infections in Denmark.

Adolescent↗
PubMed2000

Evaluation of phenotypic and genotypic methods for subtyping Campylobacter jejuni isolates from humans, poultry, and cattle.

Six methods for subtyping of Campylobacter jejuni were compared and evaluated with a collection of 90 isolates from poultry, cattle, and sporadic human clinical cases as well as from a waterborne outbreak. The applied methods were Penner heat-stable serotyping; automated ribotyping (RiboPrinting); random amplified polymorphic DNA typing (RAPD); pulsed-field gel electrophoresis (PFGE); restriction fragment length polymorphisms of the flagellin gene, flaA (fla-RFLP); and denaturing gradient gel electrophoresis of flaA (fla-DGGE). The methods were evaluated and compared on the basis of their abilities to identify isolates from one outbreak and discriminate between unrelated isolates and the agreement between methods in identifying clonal lines. All methods identified the outbreak strain. For a collection of 80 supposedly unrelated isolates, RAPD and PFGE were the most discriminatory methods, followed by fla-RFLP and RiboPrinting. fla-DGGE and serotyping were the least discriminative. All isolates included in this study were found to be typeable by each of the methods. Thirteen groups of potentially related isolates could be identified using a criterion that at least four of the methods agreed on clustering of isolates. None of the subtypes could be related to only one source; rather, these groups represented isolates from different sources. Furthermore, in two cases isolates from cattle and human patients were found to be identical according to all six methods.

Animals↗
PubMed2000

Misidentifying helicobacters: the Helicobacter cinaedi example.

Whole-cell protein electrophoresis and biochemical examination by means of a panel of 64 tests were used to identify 14 putative helicobacters to the species level. The results were confirmed by means of DNA-DNA hybridization experiments and were used to discuss misidentification of helicobacters based on 16S rRNA gene sequence data. The data indicated that comparison of near-complete 16S ribosomal DNA sequences does not always provide conclusive evidence for species level identification and may prove highly misleading. The data also indicated that "Helicobacter westmeadii" is a junior synonym of Helicobacter cinaedi and that Helicobacter sp. strain Mainz belongs to the same species. H. cinaedi occurs in various animal reservoirs, including hamsters, dogs, cats, rats, and foxes. Appropriate growth conditions and identification strategies will be required to establish the genuine significance of this widely distributed Helicobacter species.

Animals↗
PubMed1999

High-resolution genomic fingerprinting of Campylobacter jejuni and Campylobacter coli by analysis of amplified fragment length polymorphisms.

A method for high-resolution genomic fingerprinting of the enteric pathogens Campylobacter jejuni and Campylobacter coli, based on the determination of amplified fragment length polymorphism, is described. The potential of this method for molecular epidemiological studies of these species is evaluated with 50 type, reference, and well-characterised field strains. Amplified fragment length polymorphism fingerprints comprised over 60 bands detected in the size range 35-500 bp. Groups of outbreak strains, replicate subcultures, and 'genetically identical' strains from humans, poultry and cattle, proved indistinguishable by amplified fragment length polymorphism fingerprinting, but were differentiated from unrelated isolates. Previously unknown relationships between three hippurate-negative C. jejuni strains, and two C. coli var. hyoilei strains, were identified. These relationships corresponded to available epidemiological data. We conclude that this amplified fragment length polymorphism fingerprinting method may be a highly effective tool for molecular epidemiological studies of Campylobacter spp.

Animals↗
PubMed1999

Clonality of Campylobacter sputorum bv. paraureolyticus determined by macrorestriction profiling and biotyping, and evidence for long-term persistent infection in cattle.

Eighteen strains of Campylobacter sputorum bv. paraureolyticus (isolated over a 12-month period from seven dairy cows contained in a single herd) were examined by resistotyping, and macrorestriction profiling using pulsed field gel electrophoresis (PFGE). The resistotypes of these strains were identical, although repeat testing indicated resistance to metronidazole was not a reliable trait for typing purposes. Five SmaI-derived genotypes were identified among the 18 strains. In 5 of 7 cows, isolates obtained from the same animal, but from different time periods, were genotypically indistinguishable, indicating persistence of infection. Macrorestriction profiles of 5 strains representing the 5 SmaI genotypes and 8 other strains of C. sputorum from various sources, were prepared using 4 endonucleases (SmaI, SalI, BamHI and KpnI). The only other strain of C. sputorum bv. paraureolyticus examined (a Canadian isolate from human faeces), was found to have a SmaI macrorestriction profile identical with one of the five clones isolated from the cattle. Moreover, SalI and BamHI profiles of all bv. paraureolyticus strains were similar, while digestion with KpnI was not observed. By contrast, the seven strains of C. sputorum bv. sputorum yielded various macrorestriction profiles with all the enzymes used, and features distinguishing the two biovars studied could be identified. This study indicates that C. sputorum can persist in cattle for at least 12 months and exhibits a clonal population genetic structure.

Animals↗