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Folia Microbiologica Nov 2015This study provides information on the occurrence of Arcobacter in several types of water and food products of animal origin in the Czech Republic. We processed 190...
This study provides information on the occurrence of Arcobacter in several types of water and food products of animal origin in the Czech Republic. We processed 190 samples using the modified method, and the occurrence of Arcobacter spp. was confirmed in 36.8 % of these. This total incidence consisted of Arcobacter butzleri (27.3 %), Arcobacter cryaerophilus (8.4 %) and Arcobacter skirrowii (1.1 %). We newly described the common presence of Arcobacter spp. in sewage water in the Czech Republic that is released into waterways after processing in water treatment plants (86.7 %). All the acquired isolates were subject to detailed confirmation with subsequent species classification using multiplex PCR and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS). In this study, we used a modification of a method using passive filtration of an enriched sample, which could be suitable for the isolation of Arcobacter, especially in combination with Campylobacter selective agar chromogenic medium. Our studies have shown this agar to be quite suited to the isolation of Arcobacter and that it can be an appropriate instrument for accelerating culture diagnostics.
Topics: Arcobacter; Colony Count, Microbial; Culture Media; Czech Republic; Food Microbiology; Sewage
PubMed: 25912846
DOI: 10.1007/s12223-015-0395-x -
BioMed Research International 2016The genus includes species considered emerging food and waterborne pathogens. Despite has been linked to the presence of faecal pollution, few studies have...
The genus includes species considered emerging food and waterborne pathogens. Despite has been linked to the presence of faecal pollution, few studies have investigated its prevalence in wastewater, and the only isolated species were and . This study aimed to establish the prevalence of spp. at a WWTP using in parallel two culturing methods (direct plating and culturing after enrichment) and a direct detection by m-PCR. In addition, the genetic diversity of the isolates was established using the ERIC-PCR genotyping method. Most of the wastewater samples (96.7%) were positive for and a high genetic diversity was observed among the 651 investigated isolates that belonged to 424 different ERIC genotypes. However, only few strains persisted at different dates or sampling points. The use of direct plating in parallel with culturing after enrichment allowed recovering the species A. , A. , , , , , , and , most of them isolated for the first time from wastewater. The predominant species was A. , however, by direct plating predominated A. . Therefore, the overall predominance of A. was a bias associated with the use of enrichment.
Topics: Aerobiosis; Arcobacter; Bacteriological Techniques; Biodiversity; Polymerase Chain Reaction; Species Specificity; Wastewater; Water Purification
PubMed: 27981053
DOI: 10.1155/2016/8132058 -
Journal of Food Protection Mar 2003In a poultry slaughterhouse, Arcobacter contamination was examined over a period of 1 week to establish possible routes of contamination. Samples were collected from the...
In a poultry slaughterhouse, Arcobacter contamination was examined over a period of 1 week to establish possible routes of contamination. Samples were collected from the slaughter equipment and from processing water before the onset of slaughter and from the first broiler flock slaughtered on each sampling day. Characterization of 1,079 isolates by enterobacterial repetitive intergenic consensus-polymerase chain reaction and a random amplified polymorphic DNA assay resulted in the delineation of 159 Arcobacter butzleri and 139 Arcobacter cryaerophilus types. From almost all 140 neck skin samples collected before and after evisceration, A. butzleri and A. cryaerophilus were isolated simultaneously at contamination levels ranging from 10(1) to 10(4) CFU/g. Only six A. butzleri types present in the slaughterhouse environment were also present on the broiler carcasses. None of the A. cryaerophilus genotypes were detected in both the neck skin and the environmental samples. All A. butzleri types isolated from the feather samples were also isolated from broiler neck skin samples. The slaughter equipment was contaminated with arcobacters before the onset of slaughter, but it appeared unlikely that contamination through the slaughter equipment alone explained the high contamination levels on poultry products. Arcobacters were also present in processing water, but types present in water and poultry products were different. Characterization of the Arcobacter isolates did not clarify the routes of transmission, probably because of the extreme heterogeneity among Arcobacter isolates. However, the results obtained in this study brought to light insufficient decontamination at the processing plant involved in the study and confirmed the survival capacity of certain A. butzleri strains.
Topics: Abattoirs; Animals; Arcobacter; Colony Count, Microbial; Equipment Contamination; Food Contamination; Food Handling; Food Microbiology; Food-Processing Industry; Genotype; Polymerase Chain Reaction; Poultry
PubMed: 12636286
DOI: 10.4315/0362-028x-66.3.364 -
Letters in Applied Microbiology Jan 2010To evaluate the reliability of culture-independent methods in comparison with culture-dependent ones for the detection of Arcobacter spp. in estuarine waters of Southern... (Comparative Study)
Comparative Study
AIM
To evaluate the reliability of culture-independent methods in comparison with culture-dependent ones for the detection of Arcobacter spp. in estuarine waters of Southern Italy.
METHODS AND RESULTS
PCR and fluorescent in situ hybridization (FISH) procedures were used to detect arcobacters directly in water samples and after enrichment cultures. The samples totally were positive by molecular methods (PCR and FISH) but only 75% were culture positive, confirming the limitation of these latter to detect Arcobacter spp. in natural samples. Culturable arcobacters were retrieved in all times except in July, and isolated species were ascribed only to Arcobacter cryaerophilus.
CONCLUSIONS
Culturable and nonculturable forms of Arcobacter in the estuarine environment were present. PCR assays were more sensitive than traditional culture in detecting Arcobacter butzleri and A. cryaerophilus. FISH comparatively to PCR technique may provide information about cell morphology and viability of single cells.
SIGNIFICANCE AND IMPACT OF THE STUDY
Our investigation indicates the existence of an environmental reservoir of potential pathogenic arcobacters in an estuarine Italian area, which may survive under a viable but not culturable state.
Topics: Arcobacter; Colony Count, Microbial; Culture Media; DNA, Bacterial; DNA, Ribosomal; In Situ Hybridization, Fluorescence; Italy; Polymerase Chain Reaction; RNA, Bacterial; RNA, Ribosomal, 16S; Rivers; Seawater
PubMed: 19929906
DOI: 10.1111/j.1472-765X.2009.02767.x -
Microbial Pathogenesis Jan 2014In humans, arcobacters are associated with watery diarrhea and septicemia. Although, recently, more cases of diarrhea have been caused by Arcobacter species, very little...
In humans, arcobacters are associated with watery diarrhea and septicemia. Although, recently, more cases of diarrhea have been caused by Arcobacter species, very little is known about its pathogenesis. Therefore, the aim of this study was to evaluate the presence of six putative Arcobacter virulence genes (cadF, ciaB, cj1349, mviN, pldA, and tlyA), in a set of 113 Arcobacter butzleri, 40 Arcobacter cryaerophilus, and 15 Arcobacter skirrowii isolates that were recovered from various origins. The isolates were confirmed on the basis of polymerase chain reaction (PCR) amplification of genus and species specific PCR for determining three species. For confirmed isolates, PCR was carried out for the presence of virulence genes using specific primers. All A. butzleri isolates carried all six genes. For A. cryaerophilus and A. skirrowii, the cadF gene was detected just in 55 and 53.3%, ciaB in 97.5 and 86.6%, cj1349 in 45 and 60%, mviN in 90 and 80%, pldA in 32.5 and 13.3%, and tlyA in 37.5 and 40%, respectively. For A. cryaerophilus and A. skirrowii, the genes ciaB and mviN were significantly more prevalent than other virulence markers (P ≤ 0.05). The findings revealed that many of the important Arcobacter strains (86%) have these putative virulence genes which can be potential pathogenic properties for humans.
Topics: Abattoirs; Animals; Arcobacter; DNA Primers; DNA, Bacterial; Food Contamination; Food Microbiology; Genes, Bacterial; Iran; Livestock; Meat; Polymerase Chain Reaction; Virulence Factors
PubMed: 24201143
DOI: 10.1016/j.micpath.2013.10.003 -
Applied and Environmental Microbiology Mar 2004The occurrence of Arcobacter spp. was studied in seawater and plankton samples collected from the Straits of Messina, Italy, during an annual period of observation by...
The occurrence of Arcobacter spp. was studied in seawater and plankton samples collected from the Straits of Messina, Italy, during an annual period of observation by using cultural and molecular techniques. A PCR assay with three pairs of primers targeting the 16S and 23S rRNA genes was used for detection and identification of Arcobacter butzleri, Arcobacter cryaerophilus, and Arcobacter skirrowii in cultures and environmental samples. Only one of the Arcobacter species, A. butzleri, was isolated from seawater and plankton samples. With some samples the A. butzleri PCR assay gave amplified products when cultures were negative. A. cryaerophilus and A. skirrowii were never detected by culture on selective agar plates; they were detected only by PCR performed directly with environmental samples. Collectively, our data suggest that culturable and nonculturable forms of Arcobacter are present in marine environments. The assay was useful for detecting Arcobacter spp. both as free forms and intimately associated with plankton. This is the first report showing both direct isolation of A. butzleri and the presence of nonculturable Arcobacter spp. in the coastal environment of the Mediterranean Sea.
Topics: Animals; Arcobacter; Base Sequence; DNA Primers; DNA, Bacterial; DNA, Ribosomal; Genes, Bacterial; Italy; Mediterranean Sea; Plankton; Polymerase Chain Reaction; RNA, Bacterial; RNA, Ribosomal, 16S; RNA, Ribosomal, 23S; Seawater
PubMed: 15006743
DOI: 10.1128/AEM.70.3.1271-1276.2004 -
Foodborne Pathogens and Disease Mar 2010An SYBR Green real-time polymerase chain reaction (PCR) assay was developed for Arcobacter detection in food and wastewater samples. The assay was applied to 36 chicken... (Comparative Study)
Comparative Study
An SYBR Green real-time polymerase chain reaction (PCR) assay was developed for Arcobacter detection in food and wastewater samples. The assay was applied to 36 chicken and 33 wastewater samples, and the results were compared with those obtained for conventional PCR, multiplex PCR, and culture isolation. Isolates were identified by multiplex PCR and restriction fragment length polymorphism analysis of PCR-amplified DNA fragment, and typed by randomly amplified polymorphic DNA. Arcobacter sp. was detected in 25 of the 26 chicken carcasses (96%) and in 4 of the 10 liver samples (40%) by real-time PCR. Twenty-five chicken samples were positive also by conventional PCR, but in most of them the detection was only possible after 48-h enrichment. Arcobacter butzleri was the most frequently detected species. Twenty-four Arcobacter isolates were obtained from chicken samples, where A. butzleri is the only identified species. All the wastewater samples (100%) were positive for Arcobacter sp. by real-time PCR without enrichment. A. butzleri and Arcobacter cryaerophilus were detected by multiplex PCR. Fifteen samples were found to be positive by culture. Thirty-six isolates were obtained; all of them were identified as A. butzleri by multiplex PCR. However, by PCR-restriction fragment length polymorphism, 34 were identified as A. butzleri, 1 as A. cryaerophilus, and another 1 as Arcobacter skirrowii. A great genetic heterogeneity was observed by randomly amplified polymorphic DNA-PCR profiling. The real-time PCR assay developed in this work showed better detection levels than conventional PCR, together with shorter times of testing samples. Therefore, it could be used as a rapid and accurate instrument for monitoring Arcobacter contamination levels in food and water samples.
Topics: Animals; Arcobacter; Chickens; DNA, Bacterial; Foodborne Diseases; Industrial Waste; Meat; Polymerase Chain Reaction; Polymorphism, Restriction Fragment Length; Random Amplified Polymorphic DNA Technique; Sensitivity and Specificity; Water Microbiology
PubMed: 19899959
DOI: 10.1089/fpd.2009.0368 -
Journal of Global Antimicrobial... Jun 2017Arcobacter spp. are considered to be potential foodborne pathogens, and consumption of contaminated food containing these bacteria could endanger human and animal...
OBJECTIVES
Arcobacter spp. are considered to be potential foodborne pathogens, and consumption of contaminated food containing these bacteria could endanger human and animal health. Arcobacter butzleri and Arcobacter cryaerophilus are the species most frequently isolated from food of animal origin and from other samples. The aim of this study was to evaluate the susceptibility of arcobacters isolated in the Czech Republic. No information about antibiotic susceptibility and multidrug resistance of arcobacters isolated in the Czech Republic is available in the literature before now.
METHODS
The antimicrobial resistance of A. butzleri (n=80) and A. cryaerophilus (n=20) isolated from meat of animal origin, water sources and clinical samples was examined by the disk diffusion method.
RESULTS
Arcobacters were resistant to one or more antimicrobial agents in 99% (99/100) of tested isolates. Most of the Arcobacter isolates were resistant to β-lactam antibiotics, i.e. ampicillin (81.0%), amoxicillin/clavulanic acid (28.0%), cefalotin (73.0%) and aztreonam (93.0%). Arcobacters were also frequently resistant to lincosamides, i.e. clindamycin (98.0%). Of the aminoglycosides, amikacin, gentamicin and tobramycin were evaluated to be the most effective antibiotics among those tested against arcobacters.
CONCLUSIONS
These results demonstrate substantial resistance in Arcobacter isolates to 18 antimicrobial agents commonly used in medical and veterinary medicine. Multidrug resistance was found in 93.8% (75/80) of A. butzleri isolates and 70.0% (14/20) of A. cryaerophilus isolates.
Topics: Anti-Bacterial Agents; Arcobacter; Czech Republic; Disk Diffusion Antimicrobial Tests; Drug Resistance, Multiple, Bacterial; Food Microbiology; Gram-Negative Bacterial Infections; Humans; Water Microbiology
PubMed: 28400212
DOI: 10.1016/j.jgar.2017.01.006 -
Journal of Dairy Science May 2013The objectives of this study were to investigate the presence of Campylobacter spp. and Arcobacter spp. in dairy herds authorized for the production and sale of raw milk...
The objectives of this study were to investigate the presence of Campylobacter spp. and Arcobacter spp. in dairy herds authorized for the production and sale of raw milk and in a water buffalo dairy farm, and to test the antimicrobial susceptibility of the isolates. A total of 196 in-line milk filters were collected from 14 dairy farms (13 bovine and 1 water buffalo) for detection of Campylobacter spp. and Arcobacter spp. by microbiological culture. For each farm investigated, 1 isolate for each Campylobacter and Arcobacter species isolated was tested using the Etest method (AB Biodisk, Solna, Sweden) to evaluate the susceptibility to ciprofloxacin, tetracycline, chloramphenicol, ampicillin, erythromycin, and gentamicin. A total of 52 isolates were detected in 49 milk filters in 12 farms (85.7%) out of 14 and the isolates were identified as Campylobacter jejuni (6), Campylobacter hyointestinalis ssp. hyointestinalis (8), Campylobacter concisus (1), Campylobacter fetus ssp. fetus (1), Arcobacter butzleri (22), and Arcobacter cryaerophilus (14). The small number of isolates tested for antimicrobial susceptibility precludes any epidemiological consideration but highlights that all Campylobacter isolates were susceptible to macrolides, which are the first-choice drugs for the treatment of campylobacteriosis, and that resistance to fluoroquinolones and tetracycline was detected; for Arcobacter isolates, resistance to ampicillin and chloramphenicol was detected. The sale of raw milk for human consumption by self-service automatic vending machines has been allowed in Italy since 2004 and the presence of C. jejuni in in-line milk filters confirms that raw milk consumption is a significant risk factor for human infection. The high occurrence of emerging Campylobacter spp. and Arcobacter spp. discovered in dairy farms authorized for production and sale of raw milk represents an emerging hazard for human health.
Topics: Animal Husbandry; Animals; Anti-Bacterial Agents; Arcobacter; Buffaloes; Campylobacter; Campylobacter fetus; Campylobacter hyointestinalis; Campylobacter jejuni; Female; Italy; Microbial Sensitivity Tests; Milk
PubMed: 23453517
DOI: 10.3168/jds.2012-6249 -
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