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Kurt Houf - One of the best experts on this subject based on the ideXlab platform.
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Occurrence of putative virulence genes in Arcobacter species isolated from humans and animals
Journal of clinical microbiology, 2011Co-Authors: Laid Douidah, Peter Vandamme, Lieven De Zutter, Olivier Vandenberg, Anne-marie Van Den Abeele, Julie Baré, Paul De Vos, Kurt HoufAbstract:Interest in Arcobacters in veterinary and human public health has increased since the first report of the isolation of Arcobacters from food of animal origin. Since then, studies worldwide have reported the occurrence of Arcobacters on food and in food production animals and have highlighted possible transmission, especially of Arcobacter butzleri, to the human population. In humans, Arcobacters are associated with enteritis and septicemia. To assess their clinical relevance for humans and animals, evaluation of potential virulence factors is required. However, up to now, little has been known about the mechanisms of pathogenicity. Because of their close phylogenetic affiliation to the food-borne pathogen Campylobacter and their similar clinical manifestations, the presence of nine putative Campylobacter virulence genes (cadF, ciaB, cj1349, hecA, hecB, irgA, mviN, pldA, and tlyA) previously identified in the recent Arcobacter butzleri ATCC 49616 genome sequence was determined in a large set of human and animal Arcobacter butzleri, Arcobacter cryaerophilus, and Arcobacter skirrowii strains after the development of rapid and accurate PCR assays and confirmed by sequencing and dot blot hybridization.
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Arcobacter Population Dynamics in Pigs on Farrow-to-Finish Farms
Applied and environmental microbiology, 2011Co-Authors: Sarah De Smet, Peter Vandamme, Lieven De Zutter, Lies Debruyne, Frédéric Vangroenweghe, Kurt HoufAbstract:Healthy pigs are an important reservoir for the emerging human pathogen Arcobacter which can result in contamination of porcine carcasses and pork and the spread of Arcobacters into the environment. Up to now, the excretion of Arcobacters by pigs has been studied, but information about the transmission routes in fattening pigs is lacking. The present study aimed to elucidate the Arcobacter population dynamics in pigs during the fattening period on four farrow-to-finish farms. On each farm, 30 clinically healthy, 12-week-old piglets were selected. Fecal samples were collected on 10 sampling occasions until a slaughter age of 30 weeks was reached. Arcobacter spp. were isolated by a selective method and identified by multiplex PCR. The genetic diversity was examined by amplified fragment length polymorphism and enterobacterial repetitive intergenic consensus PCR. The Arcobacter presence in the fecal samples on the four farms ranged from 11.3 to 50.0%, with excretion levels of up to 10(4) CFU/g feces. The ratio in which Arcobacter species were isolated varied between the farms and over time. Characterization revealed a high degree of genotypic diversity among the isolates. Arcobacter strains persisted and spread within the finishing unit during the fattening period. The occurrence of both unique and shared genotypes in pigs in adjacent and nonadjacent pens demonstrates that transmission routes other than fecal-oral transmission occur.
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Arcobacter thereius sp. nov., isolated from pigs and ducks.
INTERNATIONAL JOURNAL OF SYSTEMATIC AND EVOLUTIONARY MICROBIOLOGY, 2009Co-Authors: Kurt Houf, Sarah De Smet, Lies Debruyne, Tom Coenye, Peter VandammeAbstract:During a Danish study on the prevalence of campylobacteria in pig abortions and food of animal origin, eight Gram-negative, slightly curved, rod-shaped, non-spore-forming bacteria were clustered by using amplified fragment length polymorphism analysis in a distinct phenon within the genus Arcobacter. In the present study, numerical analysis of whole-cell protein profiles also showed that all isolates clustered in a single group distinct from other recognized Arcobacter species. DNA-DNA hybridization among two representative strains exhibited a mean DNA-DNA relatedness value of 79 %. DNA-DNA hybridization with the type strains of recognized Arcobacter species revealed levels of DNA-DNA relatedness of 41 % or less. The DNA G+C content of the type strain was 28.5 mol%. Pairwise comparison of the 16S rRNA gene sequences with those of the type strains of established species identified Arcobacter cryaerophilus (97.9 %), Arcobacter cibarius (97.5 %) and Arcobacter skirrowii (97.2 %) as the nearest phylogenetic neighbours. The isolates could be distinguished from other Arcobacter species by means of the following biochemical tests: activities of catalase and urease, reduction of nitrate and growth on minimal medium, lack of growth at 37 degrees C under standardized aerobic and microaerobic conditions, in 4 % NaCl and 1 % glycine media. Finally, DNA fingerprints obtained by using enterobacterial repetitive intergenic consenus-PCR showed that the eight isolates represent eight strains of a single novel Arcobacter species, for which the name Arcobacter thereius sp. nov. is proposed. The type strain is LMG 24486(T) (=CCUG 56902(T)).
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Arcobacter in a food safety perspective
Archiv Fur Lebensmittelhygiene, 2009Co-Authors: Kurt HoufAbstract:Arcobacters are increasingly Isolated from food products and have been classified as emerging food pathogens In the past, they were predominantly associated with reproductive problems, mastitis and gastric ulcers in livestock, though recently they are frequently isolated from clinically healthy farm animals. In humans, Arcobacter is associated with enteritis with in some cases septicaemia. The routes of infection are Still unclear but included, person-to-person contact and consumption of contaminated water and food. At present, there is no unequivocal evidence that Arcobacters are hazards for human health, but neither are they part of the normal commensal flora. The current status Of the pathogen and the importance of food contamination is reviewed.
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survival capacity in water of Arcobacter species under different temperature conditions
Journal of Applied Microbiology, 2008Co-Authors: E Van Driessche, Kurt HoufAbstract:Aims: To assess the survival capacity in vitro of Arcobacters in water at temperatures applied in the food industry. Methods and Results: Four strains of each Arcobacter species were inoculated in potable water and water with 1% organic material and stored at 4, 7, 20, 52, 56 and 60°C. Samples were taken at known time points and the numbers of bacteria were determined on Arcobacter-selective medium. All Arcobacter species remained viable for a temperature-dependent period of time, although Arcobacter butzleri displayed a significant longer survival and heat resistance. No significant intraspecies differences were detected, resulting in no definite identification of origin or strain dependency. The survival period for all species was prolonged in the presence of the organic material only for the low temperatures. Conclusions: The present study demonstrates that water can act as a reservoir and as a potential source of Arcobacter contamination to humans and animals. Significance and Impact of the Study: This study assessed for the first time the survival of all human-related Arcobacter species in water. Particularly A. butzleri showed to be the most robust species with regard to temperature which is interesting as that species is often found in human clinical specimens.
Peter Vandamme - One of the best experts on this subject based on the ideXlab platform.
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a critical rebuttal of the proposed division of the genus Arcobacter into six genera using comparative genomic phylogenetic and phenotypic criteria
Systematic and Applied Microbiology, 2020Co-Authors: William G. Miller, Patrick J Biggs, Angela J Cornelius, Peter VandammeAbstract:Abstract The proposal to restructure the genus Arcobacter into six distinct genera was critically examined using: comparative analyses of up to 80 Epsilonproteobacterial genome sequences (including 26 Arcobacters); phylogenetic analyses of three housekeeping genes and also 342 core genes; and phenotypic criteria. Genome sequences were analysed with tools to calculate Percentage of Conserved Proteins, Average Amino-acid Identity, BLAST-based Average Nucleotide Identity, in silico DNA–DNA hybridisation values, genome-wide Average Nucleotide Identity, Alignment Fractions and G + C percentages. Genome analyses revealed the genus Arcobacter sensu lato to be relatively homogenous, and phylogenetic analyses clearly distinguished the group from other Epsilonproteobacteria. Genomic distinction of the genera proposed by Perez-Cataluna et al. [2018] was not supported by any of the measures used and a subsequent risk of strain misidentification clearly identified. Similarly, phenotypic analyses supported the delineation of Arcobacter sensu lato but did not justify the position of the proposed novel genera. The present polyphasic taxonomic study strongly supports the continuance of the classification of “aerotolerant campylobacters” as Arcobacter and refutes the proposed genus-level subdivision of Perez-Cataluna et al. [2018].
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Occurrence of putative virulence genes in Arcobacter species isolated from humans and animals
Journal of clinical microbiology, 2011Co-Authors: Laid Douidah, Peter Vandamme, Lieven De Zutter, Olivier Vandenberg, Anne-marie Van Den Abeele, Julie Baré, Paul De Vos, Kurt HoufAbstract:Interest in Arcobacters in veterinary and human public health has increased since the first report of the isolation of Arcobacters from food of animal origin. Since then, studies worldwide have reported the occurrence of Arcobacters on food and in food production animals and have highlighted possible transmission, especially of Arcobacter butzleri, to the human population. In humans, Arcobacters are associated with enteritis and septicemia. To assess their clinical relevance for humans and animals, evaluation of potential virulence factors is required. However, up to now, little has been known about the mechanisms of pathogenicity. Because of their close phylogenetic affiliation to the food-borne pathogen Campylobacter and their similar clinical manifestations, the presence of nine putative Campylobacter virulence genes (cadF, ciaB, cj1349, hecA, hecB, irgA, mviN, pldA, and tlyA) previously identified in the recent Arcobacter butzleri ATCC 49616 genome sequence was determined in a large set of human and animal Arcobacter butzleri, Arcobacter cryaerophilus, and Arcobacter skirrowii strains after the development of rapid and accurate PCR assays and confirmed by sequencing and dot blot hybridization.
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Arcobacter Population Dynamics in Pigs on Farrow-to-Finish Farms
Applied and environmental microbiology, 2011Co-Authors: Sarah De Smet, Peter Vandamme, Lieven De Zutter, Lies Debruyne, Frédéric Vangroenweghe, Kurt HoufAbstract:Healthy pigs are an important reservoir for the emerging human pathogen Arcobacter which can result in contamination of porcine carcasses and pork and the spread of Arcobacters into the environment. Up to now, the excretion of Arcobacters by pigs has been studied, but information about the transmission routes in fattening pigs is lacking. The present study aimed to elucidate the Arcobacter population dynamics in pigs during the fattening period on four farrow-to-finish farms. On each farm, 30 clinically healthy, 12-week-old piglets were selected. Fecal samples were collected on 10 sampling occasions until a slaughter age of 30 weeks was reached. Arcobacter spp. were isolated by a selective method and identified by multiplex PCR. The genetic diversity was examined by amplified fragment length polymorphism and enterobacterial repetitive intergenic consensus PCR. The Arcobacter presence in the fecal samples on the four farms ranged from 11.3 to 50.0%, with excretion levels of up to 10(4) CFU/g feces. The ratio in which Arcobacter species were isolated varied between the farms and over time. Characterization revealed a high degree of genotypic diversity among the isolates. Arcobacter strains persisted and spread within the finishing unit during the fattening period. The occurrence of both unique and shared genotypes in pigs in adjacent and nonadjacent pens demonstrates that transmission routes other than fecal-oral transmission occur.
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Arcobacter thereius sp. nov., isolated from pigs and ducks.
INTERNATIONAL JOURNAL OF SYSTEMATIC AND EVOLUTIONARY MICROBIOLOGY, 2009Co-Authors: Kurt Houf, Sarah De Smet, Lies Debruyne, Tom Coenye, Peter VandammeAbstract:During a Danish study on the prevalence of campylobacteria in pig abortions and food of animal origin, eight Gram-negative, slightly curved, rod-shaped, non-spore-forming bacteria were clustered by using amplified fragment length polymorphism analysis in a distinct phenon within the genus Arcobacter. In the present study, numerical analysis of whole-cell protein profiles also showed that all isolates clustered in a single group distinct from other recognized Arcobacter species. DNA-DNA hybridization among two representative strains exhibited a mean DNA-DNA relatedness value of 79 %. DNA-DNA hybridization with the type strains of recognized Arcobacter species revealed levels of DNA-DNA relatedness of 41 % or less. The DNA G+C content of the type strain was 28.5 mol%. Pairwise comparison of the 16S rRNA gene sequences with those of the type strains of established species identified Arcobacter cryaerophilus (97.9 %), Arcobacter cibarius (97.5 %) and Arcobacter skirrowii (97.2 %) as the nearest phylogenetic neighbours. The isolates could be distinguished from other Arcobacter species by means of the following biochemical tests: activities of catalase and urease, reduction of nitrate and growth on minimal medium, lack of growth at 37 degrees C under standardized aerobic and microaerobic conditions, in 4 % NaCl and 1 % glycine media. Finally, DNA fingerprints obtained by using enterobacterial repetitive intergenic consenus-PCR showed that the eight isolates represent eight strains of a single novel Arcobacter species, for which the name Arcobacter thereius sp. nov. is proposed. The type strain is LMG 24486(T) (=CCUG 56902(T)).
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Arcobacter cibarius sp. nov., isolated from broiler carcasses.
International Journal of Systematic and Evolutionary Microbiology, 2005Co-Authors: Kurt Houf, Jan Van Hoof, Tom Coenye, Jan Mast, Peter VandammeAbstract:Twenty Gram-negative, rod-shaped, slightly curved, non-spore-forming bacteria that gave a negative result in Arcobacter species-specific PCR tests but that yielded an amplicon in an Arcobacter genus-specific PCR test were isolated from 13 unrelated broiler carcasses. Numerical analysis of the profiles obtained by SDS-PAGE of whole-cell proteins clustered all isolates in a single group distinct from the other Arcobacter species. DNA–DNA hybridization among four representative strains exhibited DNA binding values above 91 %. DNA–DNA hybridization with reference strains of the current four Arcobacter species revealed binding levels below 47 %. The G+C contents ranged between 26·8 and 27·3 mol%. Pairwise comparison of 16S rRNA gene sequences revealed the mean values for similarity to the type strain of Arcobacter cryaerophilus (97·5 %), Arcobacter butzleri (96·5 %), Arcobacter skirrowii (96·0 %) and Arcobacter nitrofigilis (95·0 %). The levels of similarity to Campylobacter and Helicobacter species were below 88 and 87 %, respectively. The isolates could be distinguished from other Arcobacter species by the following biochemical tests: catalase, oxidase and urease activities; reduction of nitrate; growth at 25 and 37 °C under aerobic conditions; growth on 2–4 % (w/v) NaCl media; and susceptibility to cephalothin. These data demonstrate that the 20 isolates represent a single novel Arcobacter species, for which the name Arcobacter cibarius sp. nov. is proposed, with LMG 21996T (=CCUG 48482T) as the type strain.
Maria José Figueras - One of the best experts on this subject based on the ideXlab platform.
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Table_1_The Use of a DNA-Intercalating Dye for Quantitative Detection of Viable Arcobacter spp. Cells (v-qPCR) in Shellfish.DOCX
2019Co-Authors: Nuria Salas-massó, Maria José Figueras, Quyen Than Linh, Wai Hoe Chin, Anders Wolff, Karl B. Andree, Dolors M. Furones, Dang Duong BangAbstract:The genus Arcobacter (Vandamme et al., 1991), comprised of Campylobacter-related species, are considered zoonotic emergent pathogens. The presence of Arcobacter in food products like shellfish, has an elevated incidence worldwide. In this study, we developed a specific viable quantitative PCR (v-qPCR), using the dye propidium monoazide (PMA), for quantification of the viable Arcobacter spp. cells in raw oysters and mussels. The high selectivity of primers was demonstrated by using purified DNA from 38 different species, 20 of them from the genus Arcobacter. The optimization of PMA concentration showed that 20 μM was considered as an optimal concentration that inhibits the signal from dead cells at different concentrations (OD550 from 0.2 to 0.8) and at different ratios of live: dead cells (50:50 and 90:10). The v-qPCR results from shellfish samples were compared with those obtained in parallel using several culture isolation approaches (i.e., direct plating on marine and blood agar and by post-enrichment culturing in both media). The enrichment was performed in parallel in Arcobacter-CAT broth with and without adding NaCl. Additionally, the v-qPCR results were compared to those obtained with traditional quantitative (qPCR). The v-qPCR and the qPCR resulted in c.a. 94% of positive detection of Arcobacter vs. 41% obtained by culture approaches. When examining the reduction effect resulting from the use of v-qPCR, samples pre-enriched in Arcobacter-CAT broth supplemented with 2.5% NaCl showed a higher reduction (3.27 log copies) than that of samples obtained directly and those pre-enriched in Arcobacter-CAT broth isolation (1.05 and 1.04). When the v-qPCR was applied to detect Arcobacter from real shellfish samples, 15/17 samples tested positive for viable Arcobacter with 3.41 to 8.70 log copies 1g-1. This study offers a new tool for Arcobacter surveillance in seafood.
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The Use of a DNA-Intercalating Dye for Quantitative Detection of Viable Arcobacter spp. Cells (v-qPCR) in Shellfish
Frontiers Media S.A., 2019Co-Authors: Maria José Figueras, Nuria Salas-massó, Quyen Than Linh, Wai Hoe Chin, Anders Wolff, Karl B. Andree, Dolors M. Furones, Dang Duong BangAbstract:The genus Arcobacter (Vandamme et al., 1991), comprised of Campylobacter-related species, are considered zoonotic emergent pathogens. The presence of Arcobacter in food products like shellfish, has an elevated incidence worldwide. In this study, we developed a specific viable quantitative PCR (v-qPCR), using the dye propidium monoazide (PMA), for quantification of the viable Arcobacter spp. cells in raw oysters and mussels. The high selectivity of primers was demonstrated by using purified DNA from 38 different species, 20 of them from the genus Arcobacter. The optimization of PMA concentration showed that 20 μM was considered as an optimal concentration that inhibits the signal from dead cells at different concentrations (OD550 from 0.2 to 0.8) and at different ratios of live: dead cells (50:50 and 90:10). The v-qPCR results from shellfish samples were compared with those obtained in parallel using several culture isolation approaches (i.e., direct plating on marine and blood agar and by post-enrichment culturing in both media). The enrichment was performed in parallel in Arcobacter-CAT broth with and without adding NaCl. Additionally, the v-qPCR results were compared to those obtained with traditional quantitative (qPCR). The v-qPCR and the qPCR resulted in c.a. 94% of positive detection of Arcobacter vs. 41% obtained by culture approaches. When examining the reduction effect resulting from the use of v-qPCR, samples pre-enriched in Arcobacter-CAT broth supplemented with 2.5% NaCl showed a higher reduction (3.27 log copies) than that of samples obtained directly and those pre-enriched in Arcobacter-CAT broth isolation (1.05 and 1.04). When the v-qPCR was applied to detect Arcobacter from real shellfish samples, 15/17 samples tested positive for viable Arcobacter with 3.41 to 8.70 log copies 1g-1. This study offers a new tool for Arcobacter surveillance in seafood
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The Use of Two Culturing Methods in Parallel Reveals a High Prevalence and Diversity of Arcobacter spp. in a Wastewater Treatment Plant.
BioMed research international, 2016Co-Authors: Arturo Levican, Luis Collado, Maria José FiguerasAbstract:The genus Arcobacter includes species considered emerging food and waterborne pathogens. Despite Arcobacter has been linked to the presence of faecal pollution, few studies have investigated its prevalence in wastewater, and the only isolated species were Arcobacter butzleri and Arcobacter cryaerophilus. This study aimed to establish the prevalence of Arcobacter 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 Arcobacter 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. butzleri, A. cryaerophilus, Arcobacter thereius, Arcobacter defluvii, Arcobacter skirrowii, Arcobacter ellisii, Arcobacter cloacae, and Arcobacter nitrofigilis, most of them isolated for the first time from wastewater. The predominant species was A. butzleri, however, by direct plating predominated A. cryaerophilus. Therefore, the overall predominance of A. butzleri was a bias associated with the use of enrichment.
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Occurrence of potentially pathogenic Arcobacters in shellfish.
Food microbiology, 2016Co-Authors: Anna Mottola, Alba Pérez-cataluña, Maria José Figueras, Andrea Serraino, Elisabetta Bonerba, Patrizia Marchetti, Giancarlo Bozzo, Valentina Terio, Giuseppina Tantillo, Angela Di PintoAbstract:Considering that several recent cases of human gastroenteritis have been associated with species from the Arcobacter genus, and that few data are currently available about the occurrence of this genus in Italian shellfish, the aim of the present study was to evaluate the occurrence of Arcobacter spp. and the presence of virulence-associated genes. The approach consisted of cultural and biomolecular (multiplex-PCR and 16S-RFLP) methods identifying isolates, followed by PCR assays aimed at the cadF, ciaB, cjl349, irgA, hecA putative virulence genes. Arcobacter spp. was detected in 16/70 (22.8%) shellfish samples. Specifically, Arcobacter spp. was highlighted in 10/42 (23.8%) mussel and in 6/28 (21.4%) clam samples. Subsequently, biomolecular assays revealed Arcobacter butzleri in 12/16 (75%) and Arcobacter cryaerophilus 1B in 4/16 (25%) isolates. PCRs aimed at the five putative virulence genes demonstrated widespread distribution of these genes among Arcobacter isolates and some differences from the results published by other authors. Our research provides more information regarding the health risks associated with the consumption of raw bivalve molluscs and underlines the need to implement an adequate control plan by performing intensive and continuous monitoring in order to guarantee human health.
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Arcobacter ebronensis sp. nov. and Arcobacter aquimarinus sp. nov., two new species isolated from marine environment.
Systematic and applied microbiology, 2014Co-Authors: Arturo Levican, Luis Collado, Sara Rubio-arcos, Antonio J. Martínez-murcia, Maria José FiguerasAbstract:Abstract Two strains recovered from mussels (F128-2 T ) and sea water (W63 T ) were characterized as Arcobacter sp., but they could not be assigned to any known species using the molecular identification methods specific for this genus (16S rDNA-RFLP and m-PCR) and rpoB gene analysis. The 16S rRNA gene sequence similarity to the type strains of all Arcobacter species ranged from 92.2% to 96.7% with strain F128-2 T , and from 94.1% to 99.4% with strain W63 T , the most similar being A. bivalviorum (CECT 7835 T ) and A. defluvii (CECT 7697 T ), respectively. The phylogenetic analyses of 16S rRNA, and the concatenated sequences of gyrB, gyrA, rpoB, atpA and hsp60 genes confirmed that strains F128-2 T and W63 T belonged to two new lineages within the genus Arcobacter . Moreover, both strains showed differential phenotypic characteristics and MALDI-TOF mass spectra from all other Arcobacter species. Therefore, it has been demonstrated the existence of two new Arcobacter species and the proposed names are Arcobacter ebronensis (type strain F128-2 T = CECT 8441 T = LMG 27922 T ), and Arcobacter aquimarinus (type strain W63 T = CECT 8442 T = LMG 27923 T ).
Luis Collado - One of the best experts on this subject based on the ideXlab platform.
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The Use of Two Culturing Methods in Parallel Reveals a High Prevalence and Diversity of Arcobacter spp. in a Wastewater Treatment Plant.
BioMed research international, 2016Co-Authors: Arturo Levican, Luis Collado, Maria José FiguerasAbstract:The genus Arcobacter includes species considered emerging food and waterborne pathogens. Despite Arcobacter has been linked to the presence of faecal pollution, few studies have investigated its prevalence in wastewater, and the only isolated species were Arcobacter butzleri and Arcobacter cryaerophilus. This study aimed to establish the prevalence of Arcobacter 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 Arcobacter 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. butzleri, A. cryaerophilus, Arcobacter thereius, Arcobacter defluvii, Arcobacter skirrowii, Arcobacter ellisii, Arcobacter cloacae, and Arcobacter nitrofigilis, most of them isolated for the first time from wastewater. The predominant species was A. butzleri, however, by direct plating predominated A. cryaerophilus. Therefore, the overall predominance of A. butzleri was a bias associated with the use of enrichment.
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Arcobacter ebronensis sp. nov. and Arcobacter aquimarinus sp. nov., two new species isolated from marine environment.
Systematic and applied microbiology, 2014Co-Authors: Arturo Levican, Luis Collado, Sara Rubio-arcos, Antonio J. Martínez-murcia, Maria José FiguerasAbstract:Abstract Two strains recovered from mussels (F128-2 T ) and sea water (W63 T ) were characterized as Arcobacter sp., but they could not be assigned to any known species using the molecular identification methods specific for this genus (16S rDNA-RFLP and m-PCR) and rpoB gene analysis. The 16S rRNA gene sequence similarity to the type strains of all Arcobacter species ranged from 92.2% to 96.7% with strain F128-2 T , and from 94.1% to 99.4% with strain W63 T , the most similar being A. bivalviorum (CECT 7835 T ) and A. defluvii (CECT 7697 T ), respectively. The phylogenetic analyses of 16S rRNA, and the concatenated sequences of gyrB, gyrA, rpoB, atpA and hsp60 genes confirmed that strains F128-2 T and W63 T belonged to two new lineages within the genus Arcobacter . Moreover, both strains showed differential phenotypic characteristics and MALDI-TOF mass spectra from all other Arcobacter species. Therefore, it has been demonstrated the existence of two new Arcobacter species and the proposed names are Arcobacter ebronensis (type strain F128-2 T = CECT 8441 T = LMG 27922 T ), and Arcobacter aquimarinus (type strain W63 T = CECT 8442 T = LMG 27923 T ).
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Antimicrobial resistance and virulence genes of Arcobacter isolates recovered from edible bivalve molluscs
Food Control, 2014Co-Authors: Luis Collado, Ronald Jara, Néstor Vásquez, Charles TelsaintAbstract:The goals of this study were to assess the prevalence, antimicrobial susceptibility and the presence of virulence genes of Arcobacters recovered from edible bivalve molluscs. A total of 106 samples (21 clams, 18 mussels, 20 oysters, 20 razor clams, 11 scallops and 16 surf clams) were analysed by culture between 2010 and 2013. The obtained colonies were identified by multiplex PCR and PCR-RFLP and genotyped by ERIC-PCR. Furthermore, nine putative virulence genes (cadF, ciaB, cjl349, irgA, hecA, hecB, mviN, pldA and tlyA) were assessed by PCR and the antimicrobial resistance was tested by the dilution agar method. The global prevalence was 40.5%, with the highest value in surf clams (87.5%) followed by razor clams (65.0%), mussels (33.3%), clams (23.8%), scallops (18.0%) and oysters (15.0%). The most commonly found species was Arcobacter butzleri (62%) followed by Arcobacter cryaerophilus (21%), Arcobacter skirrowii (16%) and Arcobacter defluvii (1%). A high resistance was found to nalidixic acid and ampicillin, while the predominant detected virulence genes were mviN (83.8%), ciaB (82.8%) and tlyA (72.7%). Our results indicate a high prevalence of Arcobacters in shellfish and the pathogenic potential of the recovered isolates suggests that this type of food could be a plausible transmission route of virulent strains to humans.
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higher water temperature and incubation under aerobic and microaerobic conditions increase the recovery and diversity of Arcobacter spp from shellfish
Applied and Environmental Microbiology, 2014Co-Authors: Arturo Levican, Luis Collado, Clara Yustes, Carme Aguilar, M J FiguerasAbstract:Some Arcobacter species are considered emerging food-borne and waterborne pathogens, and shellfish have been suggested as one of their reservoirs. However, only a few studies have investigated the presence of Arcobacter in this kind of food. This study assesses the prevalence and diversity of Arcobacter spp. in shellfish by multiplex PCR (m-PCR) and culturing methods (under different atmospheric conditions) and evaluates the possible influence of environmental parameters (temperature, salinity, and harvesting bay). Arcobacter was detected by m-PCR and/or culturing in 61 (29.9%) of 204 shellfish samples. Of the positive samples by culturing, 41.1% were obtained under only aerobic incubation conditions, while 23.2% were obtained under only microaerobic conditions. Of 476 investigated isolates, 118 belonged to different enterobacterial repetitive intergenic consensus (ERIC)-PCR genotypes (strains) and to 11 different species. This study shows the highest diversity of Arcobacter species ever observed in samples from any origin. The most prevalent species was Arcobacter butzleri (60.2%), followed by Arcobacter molluscorum (21.2%). The prevalence of Arcobacter was significantly higher during the summer than in other seasons, being associated with an increase in water temperature. Results confirm that shellfish are a reservoir for a remarkable diversity of Arcobacter spp.
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Arcobacter bivalviorum sp. nov. and Arcobacter venerupis sp. nov., new species isolated from shellfish
Systematic and applied microbiology, 2012Co-Authors: Arturo Levican, Luis Collado, Clara Yustes, Carmen Aguilar, Ana L. Diéguez, Jesús L. Romalde, Maria José FiguerasAbstract:Abstract A group of ten Arcobacter isolates (Gram negative, slightly curved motile rods, oxidase positive) was recovered from mussels (nine) and from clams (one). These isolates could not be assigned to any known species using the molecular identification methods specific for this genus (16S rDNA-RFLP and m-PCR). The aim of this study is to establish the taxonomic position of these isolates. The 16S rRNA gene sequence similarity of mussel strain F4 T to the type strains of all other Arcobacter species ranged from 91.1% to 94.8%. The species most similar to the clams’ strain F67-11 T were Arcobacter defluvii (CECT 7697 T , 97.1%) and Arcobacter ellisii (CECT 7837 T , 97.0%). On the basis of phylogenetic analyses with 16S rRNA, rpoB , gyrB and hsp60 genes, the mussel and clam strains formed two different, new lineages within the genus Arcobacter . These data, together with their different phenotypic characteristics and MALDI-TOF mass spectra, revealed that these strains represent two new species, for which the names Arcobacter bivalviorum (type strain F4 T = CECT 7835 T = LMG 26154 T ) and Arcobacter venerupis (type strain F67-11 T = CECT 7836 T = LMG 26156 T ) are proposed.
Irene V. Wesley - One of the best experts on this subject based on the ideXlab platform.
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Arcobacter an opportunistic human food borne pathogen
2010Co-Authors: Irene V. Wesley, William G. MillerAbstract:Two recent European surveys of patient stool samples ranked Arcobacter as the fourth most frequently recovered Campylobacter-like microbe. Aerotolerant campylobacteria (Arcobacter spp.) were first described as occurring in aborted porcine and bovine fetuses. Handling and consuming raw or contaminated poultry meat are acknowledged potential sources of human Arcobacter infection. Due to the phylogenetic relationship between Campylobacter and Arcobacter, it is logical to assume that animal models, virulence factors (including adherence), invasion, cytotoxicity, and toxin production of the two organisms would be similar. Classification of Arcobacters as free-living, environmental organisms should be reflected in the gene content of the Arcobacter genomes. The genomes of some Arcobacter species would be predicted to contain niche-related genes, e.g., osmoprotectant genes in A. halophilus and nitrogen fixation genes in A. nitrofigilis. An multilocus sequence typing (MLST) method that could be used to type five Arcobacter species was recently designed. Future MLST and DNA microarray analyses will provide further insights into Arcobacter divergence. MLST analysis has identified putative horizontal gene transfer (HGT) events in Arcobacter. Members of the genus Arcobacter can be generalized as free-living organisms found predominantly in aqueous environments, occasionally associated with food animals, and infrequently associated with humans. The publication of the complete 2.341-Mb Arcobacter genome sequence has facilitated identification of unique virulence factors and genetic markers to monitor its transmission through the food chain and which underlie its distinctive epidemiology.
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Development of a Real-Time Fluorescence Resonance Energy Transfer PCR To Detect Arcobacter Species
Journal of clinical microbiology, 2007Co-Authors: Khalil Abdelbaqi, Irene V. Wesley, Valérie Prouzet-mauleon, Francis Mégraud, Alice Buissonnière, Jessica Gresser, Armelle MénardAbstract: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°C, 58.4°C, 60.6°C, and 51.8°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.
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Prevalence of Arcobacter species in market-weight commercial turkeys.
Antonie van Leeuwenhoek, 2007Co-Authors: Michelle M. E. Andersen, Irene V. Wesley, Emily Nestor, Darrell W. TrampelAbstract:The prevalence of Arcobacter in live market weight turkeys was determined for six Midwestern commercial flocks at three intervals. Samples (n = 987) were collected from cloaca, feathers, ceca, crop, drinkers and environmental samples on farms and from carcasses at slaughter. Initially, EMJH-P80 and CVA isolated Arcobacter from 7.1% (40 of 564) of samples, while Arcobacter enrichment broth and selective agar recovered the microbe in 4.7% of samples (23 of 489 samples). Although EMJH-P80 coupled with CVA yielded Arcobacter more frequently, the selectivity of the modified Arcobacter agar enhanced the recognition of Arcobacter colonies. A multiplex PCR was used to identify all Arcobacter species and to differentiate Arcobacter butzleri. The low prevalence of Arcobacter detected in cloacal swab (2.0%, 6 of 298 samples) and cecal contents (2.1%, 3 of 145 samples) suggests that Arcobacter infrequently colonizes the intestinal tract. Despite its low prevalence in live turkeys, Arcobacter spp. were identified in 93% of carcass swabs (139 of 150 samples). The overall prevalence of Arcobacter in drinker water decreased from 67% (31 of 46 samples) in the summer of 2003 to 24.7% (18 of 73 samples) during resampling in the spring of 2004 and was inversely related to the chlorination level.
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Arcobacter: An Overview
1998Co-Authors: Irene V. WesleyAbstract:Introduction The genus Arcobacter (Latin, arc-shaped bacterium) includes bacteria formerly designated Campylobacter cryaerophila (Latin; loving cold and air). They are fastidious, microaerophilic, gram-negative, spiral-shapedbacteria that are motile by means of polar flagella. Arcobacter was first isolated by Ellis et al (6) in 1977 from aborted bovine and later from porcine fetuses. Unlike other Campylobacter species, Arcobacter grows in the presence of atmospheric oxygen (aerotolerant) and at 15°/C, which is lower than temperatures used for incubation of Campylobacter (21,22). Arcobacter spp. have been associated with cases of human enteritis and septicemia (12, 13, 27, 28, 29,30); enteritis and abortion occur in livestock (6,21,22,23,24). Arcobacter spp. have been found in water, cattle, swine, poultry, and in ground pork and turkey products. Campylobacter jejuni is a major cause of human bacterial enteritis. Because of their phylogenetic similarity, the pathogenesis, distribution and routes of transmission that have been described for C. jejuni may be applicable to Arcobacter. Transmission of C. jejuni to humans occurs via consumption of contaminated undercooked poultry, water, raw milk, milk that has been contaminated after pasteurization, shellfish, and meat. Three species of Arcobacter have been recovered from man and animals: A. butzleri, A. cryaerophilus, and A. skirrowii (31,32). Of these, A. butzleri is regarded as the primary human pathogen (12). Herein we provide a review of Arcobacter and address the possibility of considering Arcobacter spp., especially A. butzleri, as emerging foodborne pathogens.
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Multiplex PCR for the identification of Arcobacter and differentiation of Arcobacter butzleri from other Arcobacters
Veterinary microbiology, 1997Co-Authors: Karen M. Harmon, Irene V. WesleyAbstract:Abstract A multiplex polymerase chain reaction (PCR) assay to identify Arcobacter isolates and to distinguish A. butzleri from other Arcobacters is described. The test uses two primer sets. Set I targets a section of the 16S rRNA genes of Arcobacter spp. Set II amplifies a portion of the 23S rRNA genes unique to A. butzleri . Specificity of the primer sets was evaluated using ATCC reference strains of A. butzleri, A. cryaerophilus, A. skirrowii, Bacteroides spp., Campylobacter spp., Helicobacter spp. and Wolinella succinogenes . Upon PCR amplification, all of the Arcobacter isolates yielded a 1223 bp product, wherease A. butzleri ATCC 49616 exhibited both a 1223 bp and a 686 bp product. No PCR product was observed for other closely related ATCC strains ( n = 37). We next analyzed by multiplex PCR field strains of Arcobacter spp. ( n = 108) which had been previously characterized to the species level by either DNA-DNA hybridization, dot blot hybridization, ribotyping or by serology. The 1223 bp multiplex PCR product identified all of the isolates as Arcobacter . The presence of both the 1223 and 686 bp amplicons identified 66 strains as A. butzleri . Speciation by multiplex PCR agreed with results obtained by the other methods. The multiplex PCR assay is specific, rapid and easy to interpret and, thus, will aid in elucidating the prevalence, epidemiology and zoonotic potential of Arcobacter .