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Journal of Food Protection Sep 2013The aim of this study was to investigate the occurrence of Arcobacter species in raw milk in Finland. A total of 177 raw milk samples, each from a separate farm, were...
The aim of this study was to investigate the occurrence of Arcobacter species in raw milk in Finland. A total of 177 raw milk samples, each from a separate farm, were examined from June to August 2011. Arcobacter species were isolated using an enrichment and selective detection procedure. Overall, 26 (15 % ) of the 177 samples yielded Arcobacter spp. Samples from 25 farms were positive for Arcobacter butzleri and from 1 farm for Arcobacter cryaerophilus. Moreover, both Arcobacter butzleri and A. cryaerophilus were recovered from 1 positive sample. To evaluate a possible genetic variability, one strain of A. butzleri from each farm and the A. cryaerophilus sample were analyzed by pulsed-field gel electrophoresis. Genotyping revealed that Arcobacter spp. populations are heterogeneous, and no dominant clone has spread in the investigated samples. Our study is the first report on the isolation of both A. butzleri and A. cryaerophilus in raw milk in Finland. Based on our findings, the presence of Arcobacter species in raw milk may pose a potential hazard for human health, in particular for consumers who prefer drinking unpasteurized milk.
Topics: Animals; Arcobacter; Cattle; Consumer Product Safety; Electrophoresis, Gel, Pulsed-Field; Finland; Food Contamination; Food Microbiology; Genotype; Humans; Milk; Polymerase Chain Reaction; Prevalence
PubMed: 23992510
DOI: 10.4315/0362-028X.JFP-13-083 -
Applied and Environmental Microbiology May 2002In this study, enterobacterial repetitive intergenic consensus PCR (ERIC-PCR) and randomly amplified polymorphic DNA PCR (RAPD-PCR) were optimized for characterization...
In this study, enterobacterial repetitive intergenic consensus PCR (ERIC-PCR) and randomly amplified polymorphic DNA PCR (RAPD-PCR) were optimized for characterization of Arcobacter butzleri, Arcobacter cryaerophilus, and Arcobacter skirrowii. In addition, a simple and rapid DNA extraction method was tested for use in both typing procedures. Both methods had satisfactory typeability and discriminatory power, but the fingerprints generated with ERIC-PCR were more reproducible and complex than those obtained with RAPD-PCR. The use of nondiluted boiled cell suspensions as DNA templates was found to be very useful in ERIC-PCR. Characterization of large numbers of Arcobacter isolates is therefore preferably performed by the ERIC-PCR procedure. Isolates for which almost identical ERIC fingerprints are generated may subsequently be characterized by RAPD-PCR, although adjustment and standardization of the amount of the DNA template are necessary. In the second part of this study, the genotypic diversity of arcobacters present on broiler carcasses was assessed by using both typing methods. A total of 228 cultures from 24 samples were examined after direct isolation and enrichment. The isolates were identified by using a multiplex PCR as A. butzleri (n = 182) and A. cryaerophilus (n = 46). A total of 131 types (91 A. butzleri types and 40 A. cryaerophilus types) were discerned without discordance between the two typing techniques. The analysis of the poultry isolates showed that poultry products may harbor not only more than one species but also multiple genotypes. All genotypes were confined to one poultry sample, and only three genotypes were found after simultaneous enrichment and direct isolation. These results demonstrate that different outcomes can be obtained in epidemiological studies depending on the isolation procedure used and the number of isolates characterized.
Topics: Arcobacter; DNA, Bacterial; Genetic Variation; Polymerase Chain Reaction; Poultry Products; Random Amplified Polymorphic DNA Technique; Reproducibility of Results
PubMed: 11976086
DOI: 10.1128/AEM.68.5.2172-2178.2002 -
Veterinary World Jun 2017This study aimed to detect putative virulence genes in species of animal and human origin.
AIM
This study aimed to detect putative virulence genes in species of animal and human origin.
MATERIALS AND METHODS
A total of 41 isolates (16 , 13 , and 12 ) isolated from diverse sources such as fecal swabs of livestock (21), raw foods of animal origin (13), and human stool samples (7) were subjected to a set of six uniplex polymerase chain reaction assays targeting putative virulence genes (, , , , , and ).
RESULTS
All the six virulence genes were detected among all the 16 isolates. Among the 13 isolates, , , , and genes were detected in 61.5, 84.6, 76.9, 76.9, 61.5, and 61.5% of isolates, respectively. Among the 12 isolates, , , , and genes were detected in 50.0, 91.6, 83.3, 66.6, 50, and 50% of isolates, respectively.
CONCLUSION
Putative virulence genes were detected in majority of the isolates examined. The results signify the potential of species as an emerging foodborne pathogen.
PubMed: 28717327
DOI: 10.14202/vetworld.2017.716-720 -
Journal of Clinical Microbiology Feb 1997Arcobacter cryaerophilus group 1B, a gram-negative, curved or helical bacillus primarily known as a bovine and porcine pathogen, was isolated from the blood of a uremic...
Arcobacter cryaerophilus group 1B, a gram-negative, curved or helical bacillus primarily known as a bovine and porcine pathogen, was isolated from the blood of a uremic patient with hematogenous pneumonia. The patient was treated successfully with ceftizoxime and tobramycin.
Topics: Aged; Bacteremia; Ceftizoxime; Drug Therapy, Combination; Female; Gram-Negative Bacteria; Gram-Negative Bacterial Infections; Humans; Pneumonia, Bacterial; Tobramycin; Uremia
PubMed: 9003624
DOI: 10.1128/jcm.35.2.489-491.1997 -
Journal of Veterinary Diagnostic... Jan 2019We applied 7 culture methods to 50 working farm dog fecal samples and 6 methods to 50 frozen home-killed raw meat diet samples to optimize recovery of a wide range of...
We applied 7 culture methods to 50 working farm dog fecal samples and 6 methods to 50 frozen home-killed raw meat diet samples to optimize recovery of a wide range of Campylobacter spp. Culture methods combined filtration, enrichment broths, and agars at 37°C and 42°C in conventional and hydrogen-enriched microaerobic atmospheres. Overall, a prevalence of 62% (31 of 50) and 6% (3 of 50) was detected in dog and meat samples, respectively, based on Campylobacter genus PCR. A total of 356 Campylobacter spp. isolates were recovered from dogs, with successful isolation by individual methods ranging from 2 to 25 dogs. The species detected most commonly were C. upsaliensis and C. jejuni, and less commonly C. coli and C. lari. Species isolated that are rarely reported from dogs included C. rectus, C. lari subsp. concheus, C. volucris, and Helicobacter winghamensis. Six isolates from dogs positive by Campylobacter genus PCR were confirmed, using 16S rRNA sequencing, as Arcobacter cryaerophilus (1) and Arcobacter butzleri (5). C. jejuni multi-locus sequence typing results revealed a diversity of sequence types in working dogs, with several uncommonly reported from other C. jejuni sources in New Zealand. Overall, 20 isolates from 3 meat samples were positive by Campylobacter genus PCR; 1 meat sample was positive for C. jejuni, 1 for C. rectus, and 1 isolate was subsequently identified as A. butzleri. The method using Campylobacter enrichment broth in a hydrogen-enriched environment on nonselective agar resulted in significantly reduced recovery of Campylobacter spp. from both sample types.
Topics: Animal Feed; Animals; Campylobacter; Campylobacter Infections; Cross-Sectional Studies; Dog Diseases; Dogs; Feces; Microbiological Techniques; Multilocus Sequence Typing; New Zealand; Polymerase Chain Reaction; Prevalence; Prospective Studies; RNA, Bacterial; RNA, Ribosomal, 16S; Sequence Analysis, RNA
PubMed: 30574836
DOI: 10.1177/1040638718820082 -
BMC Microbiology Oct 2013Bacteria belonging to the Arcobacter genus are emerging enteropathogens and potential zoonotic agents. Their taxonomy has evolved very rapidly, and there are presently... (Comparative Study)
Comparative Study
BACKGROUND
Bacteria belonging to the Arcobacter genus are emerging enteropathogens and potential zoonotic agents. Their taxonomy has evolved very rapidly, and there are presently 18 recorded species. The prevalence of species belonging to Arcobacter is underestimated because of the limitations of currently available methods for species identification.The aim of this study was to compare the performance of five PCR based methods that target regions of 16S rRNA, 23S rRNA or gyrA genes to identify Arcobacter species, and to review previous results reported in the literature using these methods.
RESULTS
The five tested methods were found not to be reliable. They misidentified between 16.8% and 67.4% of the studied strains; this was dependent upon the target regions of the tested genes. The worst results obtained were for the identification of Arcobacter cryaerophilus and Arcobacter butzleri when the 23S rRNA gene was used as the target. These species were confused with many non-targeted species.
CONCLUSION
Our results suggest that the known diversity of Arcobacter spp. in different environments could be expanded if reliable identification methods are applied in future studies.
Topics: Arcobacter; Bacteriological Techniques; DNA Gyrase; DNA, Bacterial; Molecular Diagnostic Techniques; Polymerase Chain Reaction; RNA, Ribosomal, 16S; RNA, Ribosomal, 23S
PubMed: 24090042
DOI: 10.1186/1471-2180-13-220 -
Journal of Clinical Microbiology Jan 2000The aims of this study were to investigate the prevalence of campylobacteria including Campylobacter jejuni subsp. jejuni (C. jejuni) and Campylobacter coli in human...
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.
Topics: Adolescent; Adult; Arcobacter; Bacterial Typing Techniques; Campylobacter; Campylobacter Infections; Campylobacter coli; Campylobacter jejuni; Cefoperazone; Child; Child, Preschool; Culture Media; Denmark; Deoxycholic Acid; Epsilonproteobacteria; Feces; Gram-Negative Bacteria; Helicobacter; Humans; Infant; Infant, Newborn; Middle Aged; Prevalence
PubMed: 10618103
DOI: 10.1128/JCM.38.1.286-291.2000 -
Journal of Clinical Microbiology Sep 2007A real-time PCR targeting the gyrase A subunit gene outside the quinolone resistance-determining region has been developed to detect Arcobacter species. The species...
A real-time PCR targeting the gyrase A subunit gene outside the quinolone resistance-determining region has been developed to detect Arcobacter species. The species identification was done by probe hybridization and melting curve analysis, using fluorescence resonance energy transfer technology. Discrimination between Arcobacter species was straightforward, as the corresponding melting points showed significant differences with the characteristic melting temperatures of 63.5 degrees C, 58.4 degrees C, 60.6 degrees C, and 51.8 degrees C for the Arcobacter butzleri, Arcobacter cryaerophilus, Arcobacter cibarius, and Arcobacter nitrofigilis type strains, respectively. The specificity of this assay was confirmed with pure cultures of 106 Arcobacter isolates from human clinical and veterinary specimens identified by phenotypic methods and 16S rRNA gene sequencing. The assay was then used to screen 345 clinical stool samples obtained from patients with diarrhea. The assay detected A. butzleri in four of these clinical samples (1.2%). These results were confirmed by a conventional PCR method targeting the 16S rRNA gene with subsequent sequencing of the PCR product. In conclusion, this real-time assay detects and differentiates Arcobacter species in pure culture as well as in the competing microbiota of the stool matrix. The assay is economical since only one biprobe is used and multiple Arcobacter species are identified in a single test.
Topics: Adult; Aged; Arcobacter; Bacterial Proteins; Cupressaceae; DNA Gyrase; DNA, Bacterial; DNA, Ribosomal; Diarrhea; Feces; Female; Fluorescence Resonance Energy Transfer; Gram-Negative Bacterial Infections; Humans; Male; Molecular Sequence Data; Nucleic Acid Hybridization; Polymerase Chain Reaction; RNA, Ribosomal, 16S; Sensitivity and Specificity; Sequence Analysis, DNA; Transition Temperature
PubMed: 17652482
DOI: 10.1128/JCM.00256-07 -
Journal of Food Protection Nov 2002Arcobacter, an aerotolerant Campylobacter-like organism, has been designated an emerging pathogen because of its newly recognized ability to cause diarrheal illness in... (Comparative Study)
Comparative Study
Arcobacter, an aerotolerant Campylobacter-like organism, has been designated an emerging pathogen because of its newly recognized ability to cause diarrheal illness in both humans and animals and its presence in the human food supply. Because there is no standard isolation method for its detection, the true occurrence of this pathogen is largely unknown. In addition, the lack of a standardized isolation protocol limits the ability of investigators to compare field data. Arcobacter has been detected in whole muscle and ground pork at various levels by two different isolation methods (those of deBoer and Collins). In this study, these methods were tested along with the Johnson-Murano (JM) method, developed in our laboratory. The sensitivity of each method was tested for ground pork inoculated with Arcobacter butzleri and Arcobacter cryaerophilus IA at levels of 10(4), 10(3), 10(2), and 10(1) CFU/g. Controls included tubes with uninoculated pork and broth tubes without pork. All samples that were morphologically similar to Arcobacter were analyzed by Gram staining and by catalase and oxidasereactions. Presumptive positive samples were confirmed by the polymerase chain reaction. The JM method was determined to be the most sensitive, detecting A. butzleri down to a level of 10(1) CFU/g in 100% of the samples and detecting A. cryaerophilus IA at a level of 10(1) CFU/g in 75% of samples. In a pure buffer system, the Collins method was as effective as the JM method in isolating both organisms to levels of 10(1) cells per g.
Topics: Animals; Arcobacter; Catalase; Colony Count, Microbial; Culture Media; Food Contamination; Food Microbiology; Gentian Violet; Meat Products; Oxidoreductases; Phenazines; Polymerase Chain Reaction; Sensitivity and Specificity; Swine
PubMed: 12430704
DOI: 10.4315/0362-028x-65.11.1784 -
Microbiology and Immunology 2007We have recently developed a cytolethal distending toxin (cdt) gene-based species-specific multiplex PCR assay for identifying Campylobacter jejuni, C. coli and C....
Evaluation of a cytolethal distending toxin (cdt) gene-based species-specific multiplex PCR assay for the identification of Campylobacter strains isolated from poultry in Thailand.
We have recently developed a cytolethal distending toxin (cdt) gene-based species-specific multiplex PCR assay for identifying Campylobacter jejuni, C. coli and C. fetus. In the present study, the applicability of this assay was evaluated with 34 Campylobacter-like organisms isolated from poultry in Thailand for species identification and was compared with other assays including API Campy, 16S rRNA gene sequence, and hippuricase (hipO) gene detection. Of the 34 strains analyzed, 20, 10 and 1 were identified as C. jejuni, C. coli, and Arcobacter cryaerophilus, respectively, and 3 could not be identified by API Campy. However, 16S rRNA gene analysis, showed that all 34 strains are C. jejuni/coli. To discriminate between these 2 species, the hipO gene, which is specifically present in C. jejuni, was examined by PCR and was detected in 20 strains, which were identified as C. jejuni by API Campy but not in the remaining 14 strains. Collective results indicated that 20 strains were C. jejuni whereas the 14 strains were C. coli. When the cdt gene-based multiplex PCR was employed, however, 19, 20 and 19 strains were identified as C. jejuni while 13, 14 and 13 were identified as C. coli by the cdtA, cdtB and cdtC gene-based multiplex PCR, respectively. Pulsed-field gel electrophoresis revealed that C. jejuni and C. coli strains analyzed are genetically diverse. Taken together, these data suggest that the cdt gene-based multiplex PCR, particularly cdtB gene-based multiplex PCR, is a simple, rapid and reliable method for identifying the species of Campylobacter strains.
Topics: Animals; Bacterial Toxins; Bacterial Typing Techniques; Campylobacter; Campylobacter Infections; DNA Primers; Electrophoresis, Gel, Pulsed-Field; Genes, Bacterial; Genetic Variation; Polymerase Chain Reaction; Poultry; Poultry Diseases; Thailand
PubMed: 17895609
DOI: 10.1111/j.1348-0421.2007.tb03974.x