Arcobacter Skirrowii Research Articles

Meta-analysis on the global prevalence of Arcobacter in food-producing animals and humans

The genus Arcobacter has been associated with illnesses in both animals and humans, where Arcobacter butzleri, Arcobacter cryaerophilus, and Arcobacter skirrowii have been linked to numerous cases of gastrointestinal diseases in humans. While isolated instances of Arcobacter infection have been reported in certain areas, comprehensive data reflecting the global impact of Arcobacter infection are lacking. This meta-analysis was conducted with the objective of assessing the aggregated prevalence of Arcobacter across diverse sources on a global scale. We conducted a thorough literature search of the Scopus, PubMed, and ScienceDirect databases to identify studies published from 1992 to 2022 on Arcobacter prevalence in humans and food-producing animals. We utilized multilevel random effects meta-analysis models to gauge the average occurrence of Arcobacter and to examine various factors that could influence incidence outcomes. Seventy-five articles were included in the meta-analysis. The pooled prevalence of Arcobacter spp. from different sources was 21.9% (95% CI: 18.0%–26.1%), and the mean prevalence of A. butzleri, A. cryaerophilus, and A. skirrowii was 15.1%, 2.8%, and 0.1%, respectively. Arcobacter spp. had the lowest prevalence in humans (1.8%; 95% CI: 0.7%–3.3%) and the highest in broilers (38.8%; 95% CI: 28.0%–50.1%). Among animal-derived food products, carcasses or carcass parts exhibited the highest Arcobacter spp. prevalence of 28.6% (28.6%; 95% CI: 23.7%–33.7%). This meta-analysis revealed that A. butzleri is the most prevalent Arcobacter species worldwide, with broilers, as well as seafood, being the primary hosts of Arcobacter spp. We recommend developing appropriate prevention strategies and conducting further local in-depth studies to establish the actual epidemiological burden of Arcobacter.

Determination of the presence and antimicrobial resistance of Arcobacter species in broiler carcasses at different stages of slaughter line.

In this study, to investigate Arcobacter spp. contamination post-scalding and de-feathering, post-evisceration, post-chilling, and packaged products, which are the most essential contamination stages of broiler slaughter, a total of 108 samples were taken from three different broiler slaughterhouses at different times. Isolates obtained by cultural methods in 104 of 108 samples were analyzed by mPCR method to identify pathogen Arcobacter spp. Arcobacter butzleri, Arcobacter cryaerophilus, and mixed contamination of both Arcobacter species were detected in 51 samples. Of the 51 isolates, 27 (52.9%) were A. butzleri, 16 (31.4%) were A. cryaerophilus, and 8 (15.7%) were mixed contamination of A. butzleri and A. cryaerophilus, while Arcobacter skirrowii was not detected. A. butzleri and A. cryaerophilus contamination was 59.2% post-scalding and de-feathering, 43.4% post-evisceration, 44.4% and 48.1% post-chilling and in packaged products, respectively. All A. butzleri strains were found to be 100% resistant to cefoperazone and penicillin and sensitive to tetracycline. A. cryaerophilus strains were 100% resistant to cefoperazone, penicillin, and cloxacillin and susceptible to tetracycline and erythromycin. In the study, it was determined that Arcobacter spp. caused a very intense contamination (85.18%-100%) and also contamination rates of identified pathogen strains (A. butzleri and A. cryaerophilus) were very high (59.2% and 43.4%) in broiler slaughtering stages. Considering that each step in broiler slaughter could contaminate the next stage, developing a safe slaughter and minimizing the risk toward the final product, it was concluded that critical control points could not be well managed in broiler slaughterhouses, and broiler meat may pose a significant risk to public health.

Comparison of Two Culture-Based Detection Systems for the Isolation of Arcobacter butzleri, Arcobacter cryaerophilus, and Arcobacter skirrowii in Raw Ground Poultry

Arcobacters are emerging pathogens that have been underestimated due to a lack of a standardized isolation method. The aim of this research was to evaluate the ability to isolate Arcobacter butzleri, Arcobacter cryaerophilus, and Arcobacter skirrowii using two Arcobacter-specific culture detection systems: (i) the Houf broth and modified charcoal cefoperazone deoxycholate agar supplemented with cefoperazone, amphotericin B, and teicoplanin (HB/mCCDA+CAT), and (ii) the Nguyen-Restaino-Juárez Arcobacter enrichment broth and chromogenic agar (NRJ-B/M). Both detection systems were evaluated for productivity ratio, sensitivity, and specificity. As a result, the productivity ratio for both plating agars were >90%, which indicates that the selective agents used in the two plating agars did not inhibit Arcobacter growth. Moreover, sensitivity evaluations using artificially inoculated retail ground poultry (n = 780) determined that both detection systems were able to isolate A. butlzeri with >95% sensitivity at the 0.1 and 1.0–2.0 CFU/g detection level. The sensitivity in A. cryaerophilus isolation was higher for NRJ-B/M (78.0% at 0.1 CFU/g; 95.1% at 1.0–2.0 CFU/g) when compared with HB/mCCDA+CAT (34.1% at 0.1 CFU/g; 51.2% at 1.0–2.0 CFU/g). Both detection systems resulted in <50% sensitivity when isolating A. skirrowii at 0.1 and 1.0–2.0 CFU/g; however, the sensitivity for NRJ-B/M was significantly higher than HB/mCCDA+CAT. At the detection level of 5.0 CFU/g, both detection systems were able to isolate A. skirrowii with 100% sensitivity. Specificity comparisons using uninoculated ground poultry samples (n = 40) indicated the growth of background microbiota were significantly inhibited or could be easily differentiated on NRJ-B/M (90.0%, specificity) when compared with HB/mCCDA+CAT (30.0%, specificity). Overall, these results show that the NRJ-B/M detection system is a more sensitive and specific detection system when isolating Arcobacter spp. from ground chicken.

Genetic characteristics, antimicrobial resistance, and prevalence of Arcobacter spp. isolated from various sources in Shenzhen, China.

Arcobacter spp. is a globally emerging zoonotic and foodborne pathogen. However, little is known about its prevalence and antimicrobial resistance in China. To investigate the prevalence of Arcobacter spp. isolated from various sources, 396 samples were collected from human feces, chicken cecum, and food specimens including chicken meat, beef, pork, lettuce, and seafood. Arcobacter spp. was isolated by the membrane filtration method. For 92 strains, the agar dilution method and next-generation sequencing were used to investigate their antimicrobial resistance and to obtain whole genome data, respectively. The virulence factor database (VFDB) was queried to identify virulence genes. ResFinder and the Comprehensive Antibiotic Resistance Database (CARD) were used to predict resistance genes. A phylogenetic tree was constructed using the maximum likelihood (ML) method with core single-nucleotide polymorphisms (SNPs). We found that 27.5% of the samples (n = 109) were positive for Arcobacter spp., comprising Arcobacter butzleri (53.0%), Arcobacter cryaerophilus (39.6%), and Arcobacter skirrowii (7.4%). Chicken meat had the highest prevalence (81.2%), followed by seafood (51.9%), pork (43.3%), beef (36.7%), lettuce (35.5%), chicken cecum (8%), and human fecal samples (0%, 0/159). Antimicrobial susceptibility tests revealed that 51 A. butzleri and 40 A. cryaerophilus strains were resistant to streptomycin (98.1, 70%), clindamycin (94.1, 90%), tetracycline (64.7, 52.5%), azithromycin (43.1%, 15%), nalidixic acid (33.4, 35%), and ciprofloxacin (31.3, 35%) but were susceptible to erythromycin, gentamicin, chloramphenicol, telithromycin, and clindamycin (≤10%). A. skirrowii was sensitive to all experimental antibiotics. The virulence factors tlyA, mviN, cj1349, ciaB, and pldA were carried by all Arcobacter spp. strains at 100%, and the following percentages were cadF (95.7%), iroE (23.9%), hecB (2.2%), hecA, and irgA (1.1%). Only one A. butzleri strain (F061-2G) carried a macrolide resistance gene (ereA). One A. butzleri and one A. cryaerophilus harbored resistance island gene clusters, which were isolated from pork and chicken. Phylogenetic tree analysis revealed that A. butzleri, A. cryaerophilus, and A. skirrowii were separated from each other. To our knowledge, this is the first report of the isolation of Arcobacter spp. from vegetables and seafood in China. The resistance island gene cluster found in pork and chicken meat and the presence of virulence factors could be a potential risk to human health.

A New Method for Detection of Arcobacter butzleri, Arcobacter cryaerophilus, and Arcobacter skirrowii Using a Novel Chromogenic Agar

Arcobacter species are gram-negative rods that have been implicated in food- and waterborne illness. Although various cultural isolation methods have been proposed, the current procedures are unable to fully suppress the growth of background microbiota present in food samples, which inhibits Arcobacter isolation. The purpose of this study was to develop a selective enrichment broth and chromogenic plating medium to detect three Arcobacter species that have been recognized as emerging foodborne pathogens: Arcobacter butzleri, Arcobacter cryaerophilus, and Arcobacter skirrowii. The developed Nguyen-Restaino-Juárez (NRJ) Arcobacter detection system consists of a selective enrichment broth (NRJ-B) and a selective-differential plating medium (NRJ-M). The protocol of the detection method was determined by evaluating the growth of A. butzleri, A. cryaerophilus, and A. skirrowii under various temperatures (30, 35, and 42°C) and incubation (aerobic, microaerophilic, and anaerobic) conditions. Additionally, 47 Arcobacter strains and 39 non-Arcobacter strains were tested in inclusivity and exclusivity evaluations of NRJ-B and NRJ-M. Overall, the study determined that the optimal growth conditions of Arcobacter species using the NRJ Arcobacter detection system were aerobic incubation at 30°C. NRJ-B supported good growth of A. butzleri, A. cryaerophilus, and A. skirrowii while effectively suppressing the growth of non-Arcobacter strains after 48 h. Furthermore, NRJ-M yielded 97.8% inclusivity and 100.0% exclusivity using the tested strains and resulted in salmon-pigmented Arcobacter colonies (1.0 to 1.5 mm in diameter) after 72 h. The novel protocol is the first to develop a chromogenic plating medium for the isolation of Arcobacter species. This simple and accurate test method would greatly contribute to understanding the distribution of pathogenic Arcobacter species in food samples.

Antimicrobial resistance and virulence genes profiles of Arcobacter butzleri strains isolated from back yard chickens and retail poultry meat in Chile.

This research aims to investigate the presence and pathogenic potential of Arcobacter in poultry meat samples purchased in the retail market of Valdivia (South of Chile) as well as in faecal samples from backyard chickens from rural areas around this city. The isolates obtained were identified by molecular methods. Furthermore, putative virulence genes were assessed by PCR and the antimicrobial resistance was tested by phenotypic methods. Arcobacter was present in 41·6% of the samples, with the highest value in retail poultry meat (55·7%) followed by backyard production (28·0%). Arcobacter butzleri was the most prevalent species (75·6%) followed by Arcobacter skirrowii (14·8%) and Arcobacter cryaerophilus (9·6%). An 8·5% of A. butzleri strains from meat were resistant to both ciprofloxacin and tetracycline and 6·1% were resistant to erythromycin, while none was resistant to gentamycin, unlike strains from domestic chickens, which showed no resistance. Furthermore, A. butzleri strains from chicken meat presented a higher prevalence of virulence genes than strains from domestic chickens. In fact, in this last group, some genes (hecA, hecB and irgA) were completely absent. Therefore, this study provides insight on the epidemiology of Arcobacter in Chilean poultry and suggests that under traditional breeding conditions strains are, apparently, less pathogenic and drug resistant.

Isolation of Arcobacter spp. and identification of isolates by multiplex PCR from various domestic poultry and wild avian species

PurposeThe purpose of the present study was to determine the extent and seasonal prevalence of Arcobacter spp. in domestic poultry and wild birds in the Kars region of Turkey using multiplex polymerase chain reaction (m-PCR).MethodsIn this study, 1570 samples were collected from domestic poultry and wild avian species. The numbers of collected samples were as follows: 182 fecal samples from chickens, geese, and turkeys from family farms in the Kars region in Turkey; 1089 cloacal swab samples from chickens, geese, ducks, turkeys, and quails from family farms in this region; and 299 fecal samples from wild pigeons, crows, and owls in the same region.ResultsArcobacter spp. were isolated from 17.43%, 35.77%, 3.63%, 6.87%, and 3.33% of the cloacal swab samples obtained from geese, ducks, chickens, turkeys, and quails, respectively. In the stool samples, Arcobacter spp. were isolated from 9.62%, 13.33%, and 4% of chicken, goose, and turkey samples, respectively. In wild birds, the isolation rates of Arcobacter spp. were 6.6%, 12.15%, and 0% in pigeons, crows, and owls, respectively. Using m-PCR, among 171 Arcobacter spp. isolates obtained from poultry and wild birds, 67, 78, 24, and 2 were identified as Arcobacter cryaerophilus, Arcobacter butzleri, Arcobacter skirrowii, and Arcobacter cibarius, respectively.ConclusionsBoth poultry and wild avian species exhibited variable rates of Arcobacter species positivity. The presence of Arcobacter spp. in the digestive tracts of healthy poultry and wild birds may serve as a potential reservoir for the dissemination of these microbes in the environment and their transmission to other animals and humans.

Freshwater Sediment Microbial Communities Are Not Resilient to Disturbance From Agricultural Land Runoff.

Microorganisms are critically important for the function of surface water ecosystems but are frequently subjected to anthropogenic disturbances at either acute (pulse) or long-term (press) scales. Response and recovery of microbial community composition and function following pulse disturbance is well-studied in controlled, laboratory scale experiments but is less well-understood in natural environments undergoing continual press disturbance. The objectives of this study were to determine the drivers of sediment microbial compositional and functional changes in freshwaters receiving continual press disturbance from agricultural land runoff and to evaluate the ability of the native microbial community to resist disturbance related changes as a proxy for freshwater ecosystem health. Freshwater sediments were collected seasonally over 1 year in Kewaunee County, Wisconsin, a region impacted by concentrated dairy cattle farming, manure fertilization, and associated agricultural runoff which together serve as a press disturbance. Using 16S rRNA gene amplicon sequencing, we found that sediments in locations strongly impacted by intensive agriculture contain significantly higher abundances (p < 0.01) of the genera Thiobacillus, Methylotenera, Crenotrhix, Nitrospira, and Rhodoferax compared to reference sediments, and functions including nitrate reduction, nitrite reduction, and nitrogen respiration are significantly higher (p < 0.05) at locations in close proximity to large farms. Nine species-level potential human pathogens were identified in riverine sediments including Acinetobacer lwoffi and Arcobacter skirrowii, two pathogens associated with the cattle microbiome. Microbial community composition at locations in close proximity to intensive agriculture was not resistant nor resilient to agricultural runoff disturbance within 5 months post-disturbance but did reach a new, stable microbial composition. From this data, we conclude that sediment microbial community composition is sensitive and shifts in response to chemical and microbial pollution from intensive agriculture, has a low capacity to resist infiltration by non-native, harmful bacteria and, overall, the natural buffering capacity of freshwater ecosystems is unable to fully resist the impacts from agricultural press disturbance.

The Investigation of the Presence and Antimicrobial Profiles of Arcobacter Species in Sheep Carcasses and Feces

DOI : 10.26650/actavet.2019.18007 This study was designed to investigate the presence and the prevalence of Arcobacter butzleri, Arcobacter cryaerophilus, and Arcobacter skirrowii in sheep carcass swabs and feces from sheep with and without diarrhea raised in Istanbul. Also it was aimed to determine their resistance profiles against some antimicrobials. For this purpose, 50 fecal samples from sheep without diarrhea, 50 fecal samples from sheep with diarrhea, and 50 carcass swab samples from sheep were the material of this study. Arcobacter spp. were isolated from 49 (32.6%) of total 150 samples. 34 samples (68%) of 50 fecal samples from sheep with diarrhea, 5 samples (10%) from 50 fecal samples of sheep without diarrhea and 10 samples (20%) of 50 sheep carcass swabs were found to be positive according to isolation results. According to multiplex Polymerase Chain Reaction (mPCR) results, 31 of 49 were identified as A. skirrowii (63.3%), 9 of 49 were A. butzleri (18.3%), while 9 of 49 were A. cryaerophilus (18.3%). As a conclusion, Arcobacter species should be taken into consideration especially in sheep with diarrhea. Additionally, it should be considered that arcobacters have started to gain resistance against fluoroquinolones. Cite this article as : Celik, C., Ikiz, S., 2019. The Investigation of the Presence and Antimicrobial Profiles of Arcobacter Species in Sheep Carcasses and Feces. Acta Vet Eurasia 2019; DOI: 10.26650/actavet.2019.18007

Antimicrobial Susceptibility and Genomic Structure of Arcobacter skirrowii Isolates

Campylobacter spp. are considered the most common bacterial cause of foodborne gastroenteritis in the world. The family Campylobacteraceae includes the genus Arcobacter with the three species Arcobacter butzleri, Arcobacter cryaerophilus, and Arcobacter skirrowii as emergent enteropathogens and potential zoonotic agents. Here, we characterized genome sequences of Arcobacter that were isolated from water poultry on farms in Germany. Isolates were cultured, identified by MALDI-TOF MS and identification was verified with PCR assays. Antibiotic susceptibility testing of isolates was carried out with erythromycin, ciprofloxacin, doxycycline, tetracycline, gentamicin, and streptomycin using the gradient strip method (E-test). We also sequenced whole genomes and predicted antibiotic resistance determinants, virulence factors, performed a phylogenetic analysis to determine the genetic relatedness of these isolates and searched for plasmids.

Complete Genome Sequence of the Arcobacter skirrowii Type Strain LMG 6621.

Arcobacter skirrowii is a species of veterinary importance, originally recovered from the feces, aborted fetuses, and preputial fluids of livestock. We present here the whole-genome sequence of the A. skirrowii type strain LMG 6621 (= 449/80T = CCUG 10374T), isolated in the United Kingdom from a lamb diarrheal fecal sample.

Genotypic Characterization of Arcobacter spp. Isolated from Chicken Meat in Brazil.

Arcobacter butzleri, Arcobacter cryaerophilus, and Arcobacter skirrowii are Gram-negative pathogenic microorganisms that cause watery diarrhea and septicemia in humans. The aims of this study were to detect the presence of Arcobacter spp. in chicken meat from butcher shops in São Paulo (Brazil) and to verify their virulence genes and genotypic profiles. A total of 300 chicken cuts were analyzed. The results show the presence of Arcobacter spp. in 18.3% of samples, which were identified as A. butzleri (63.6%) and A. cryaerophilus (36.3%). All strains were positive for the virulence genes ciaB and mviN, followed by cj1349 (98%), pldA (94.4%), cadF (72.7%), tlyA (92.7%), hecA (49%), irgA (47.2%), and hecB (34.5%). These strains were subjected to single-enzyme amplified fragment length polymorphism. Nineteen genotypic profiles were obtained for A. butzleri, and 17 for A. cryaerophilus. These results confirm the presence of virulent strains of A. butzleri and A. cryaerophilus in the chicken meat in Brazil. The presence of potentially virulent strains of Arcobacter highlights a possible public health risk, particularly with respect to ingestion of undercooked foods and cross-contamination from uncooked foods during food preparation and contaminated utensils.

Occurrence of Arcobacter species in animal faeces, foods of animal origin and humans in Andhra Pradesh, India

A total of n=510 samples comprising of faecal swabs from livestock (n=245), foods of animal origin (n=180) and human stools (n=85) were examined to detect Arcobacter species employing cultural and PCR methods. Overall, 11.7% of samples were found positive for Arcobacter species by genus-specific PCR. On screening faecal samples of livestocks, higher positivity was observed in pigs (23.3%), followed by chicken (16.6%), turkey (15.0%), cattle (10.0%), duck (10.0%) and sheep (6.6%). With regard to foods of animal origin, higher positivity was observed in pork samples (15.0%), followed by chicken (12.5%), milk (10.0%) and mutton (7.5%). In humans, the stool samples of farm workers revealed high positivity (13.3%) followed by veterinary students (8.0%) and diarrhoeic humans (6.66%). Using cultural method, a total of 41 (8.03%) Arcobacter isolates were recovered. The multiplex-PCR assay enabled detection of Arcobacter butzleri (16/41), Arcobacter cryaerophilus (13/41) and Arcobacter skirrowii (12/41). The results signify the occurrence of Arcobacter species in animal faeces, foods of animal origin and humans in Andhra Pradesh, India.

Virulence gene profiles of Arcobacter species isolated from animals, foods of animal origin, and humans in Andhra Pradesh, India.

Aim::This study aimed to detect putative virulence genes in Arcobacter species of animal and human origin.Materials and Methods::A total of 41 Arcobacter isolates (16 Arcobacter butzleri, 13 Arcobacter cryaerophilus, and 12 Arcobacter skirrowii) 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 Arcobacter putative virulence genes (ciaB, pldA, tlyA, mviN, cadF, and cj1349).Results::All the six virulence genes were detected among all the 16 A. butzleri isolates. Among the 13 A. cryaerophilus isolates, cadF, ciaB, cj1349, mviN, pldA, and tlyA genes were detected in 61.5, 84.6, 76.9, 76.9, 61.5, and 61.5% of isolates, respectively. Among the 12 A. skirrowii isolates, cadF, ciaB, cj1349, mviN, pldA, and tlyA 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 Arcobacter isolates examined. The results signify the potential of Arcobacter species as an emerging foodborne pathogen.

Comparative isolation and genetic diversity of Arcobacter sp. from fish and the coastal environment.

Arcobacter sp. are emerging food- and water-borne human pathogens. In this study, comparison of two selective media suggested Arcobacter blood agar to be more efficient in yielding Arcobacter sp. from seafood. Furthermore, the isolation of Arcobacter sp. such as Arcobacter butzleri, A. skirrowii, A. mytili and A. defluvii from seafood suggests diverse sources of contamination of seafood by Arcobacter sp. Analysis of enterobacterial repetitive intergenic consensus sequence-PCR patterns of A. butzleri showed high genetic diversity and lack of clonality among the isolates. Arcobacter contamination of seafood is an emerging issue both from seafood safety and seafood trade point of view.

Occurence and antimicrobial resistance of Arcobacter species in food and slaughterhouse samples

The objectives of this study were i) to isolate Arcobacter species (Arcobacter butzleri, Arcobacter skirrowii, Arcobacter cryaerophilus) from different foods and sources, ii) to verify the isolates by multiplex PCR assay, iii) to detect the antibiotic resistance profiles of the isolates. In this study a total of 60 Arcobacter isolates were obtained. Arcobacter species were mostly isolated from swab samples (40%), followed by wastewater (29.1%), broiler wing meat (30%), raw milk (23.9%) and minced meat (6.6%). Regarding the seasonal distribution of Arcobacter from swab and wastewater samples, the bacterium was commonly isolated from wastewater in winter and spring, while it was frequently detected in swab samples during autumn and spring. All of the isolates were found to be resistant to nalidixic acid, ampicillin, rifampin, and erythromycin. The most effective antibiotic was tetracycline, because 96.66% of the isolates were susceptible against it. This is the first report of the isolation, seasonal distribution and antimicrobial susceptibility of Arcobacter species in cattle slaughterhouse samples in Turkey. These results indicate that foods of animal origin and cattle slaughterhouses are significant source of the antimicrobial resistant arcobacters.

Development and evaluation of multiplex PCR assays for rapid detection of virulence-associated genes in Arcobacter species

As the pathogenicity of Arcobacter species might be associated with various virulence factors, this study was aimed to develop and optimize three single-tube multiplex PCR (mPCR) assays that can efficiently detect multiple virulence-associated genes (VAGs) in Arcobacter spp. including the Arcobacter butzleri, Arcobacter cryaerophilus and Arcobacter skirrowii, respectively. The recognized target virulence factors used in the study were fibronectin binding protein (cj1349), filamentous hemagglutinin (hecA), hemolysin activation protein (hecB), hemolysin (tlyA), integral membrane protein virulence factor (mviN), invasin (ciaB), outer membrane protein (irgA) and phospholipase (pldA). Identical results were obtained between singleplex PCR and mPCR assays and no cross- and/or non-specific amplification products were obtained when tested against other closely related bacterial species. The sensitivities of these three mPCR assays were ranging from 1ngμL−1 to 100ngμL−1 DNA. The developed assays with combinations of duplex or triplex PCR primer pairs of VAGs were further evaluated and validated by applying them to isolates of the A. butzleri, A. cryaerophilus and A. skirrowii recovered from fecal samples of human and animal origins. The findings revealed that the distribution of the ciaB (90%), mviN (70%), tlyA (50%) and pldA (45%) genes among these target species was significantly higher than the hecA (16%), hecB (10%) and each of irgA and cj1349 (6%) genes, respectively. The newly developed mPCR assays can be used as rapid technique and useful markers for the detection, prevalence and profiling of VAGs in the Arcobacter spp. Moreover, these assays can easily be performed with a high throughput to give a presumptive identification of the causal pathogen in epidemiological investigation of human infections.

The Use of Two Culturing Methods in Parallel Reveals a High Prevalence and Diversity of Arcobacter spp. in a Wastewater Treatment Plant.

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.

Prevalence of ten putative virulence genes in the emerging foodborne pathogen Arcobacter isolated from food products

Arcobacter spp. are considered to be emerging food- and waterborne pathogens for both humans and animals. However, their virulence mechanisms are still poorly understood. In this study the presence of ten virulence genes (cadF, ciaB, cj1349, hecA, hecB, mviN, pldA, irgA, tlyA and iroE) was assessed in a set of 47 strains of Arcobacter butzleri, 10 of Arcobacter cryaerophilus and 1 Arcobacter skirrowii strain recovered from different food products (pork, chicken, beef, milk, clams and mussels). Overall, the genes cadF, ciaB, cj1349, mviN, pldA and tlyA were detected in all A. butzleri and A. skirrowii strains. Lower detection rates were observed for irgA, iroE, hecA and hecB. The genes hecB and iroE were detected neither in A. cryaerophilus nor in A. skirrowii. The genes hecA and irgA were not detected in A. skirrowii. It was noteworthy that the genes hecA and hecB were significantly (P < 0.05) highly detected in A. butzleri strains isolated from clams compared with strains isolated from milk and chicken. Therefore, our findings underline clams as a source of A. butzleri strains with high prevalence of putative virulence genes. This could be hazardous to human health, especially because these bivalves are usually consumed raw or undercooked.

Arcobacter butzleri, Arcobacter cryaerophilus, and Arcobacter skirrowii Circulation in a Dairy Farm and Sources of Milk Contamination.

Even though dairy cows are known carriers of Arcobacter species and raw or minimally processed foods are recognized as the main sources of human Arcobacter infections in industrialized countries, data on Arcobacter excretion patterns in cows and in milk are scant. This study aimed to identify potentially pathogenic Arcobacter species in a dairy herd and to investigate the routes of Arcobacter transmission among animals and the potential sources of cattle infection and milk contamination. A strategy of sampling the same 50 dairy animals, feed, water, and milk every month for a 10-month period, as well as the sampling of quarter milk, animal teats, the milking environment, and animals living on the farm (pigeons and cats), was used to evaluate, by pulsed-field gel electrophoresis (PFGE), the characteristic patterns in animals, their living environment, and the raw milk they produced. Of the 463 samples collected, 105 (22.6%) were positive for Arcobacter spp. by culture examination. All the matrices except quarter milk and pigeon gut samples were positive, with prevalences ranging from 15 to 83% depending on the sample. Only three Arcobacter species, Arcobacter cryaerophilus (54.2%), A. butzleri (34.2%), and A. skirrowii (32.3%), were detected. PFGE analysis of 370 isolates from positive samples provided strong evidence of Arcobacter circulation in the herd: cattle likely acquire the microorganisms by orofecal transmission, either by direct contact or from the environment, or both. Water appears to be a major source of animal infection. Raw milk produced by the farm and collected from a bulk tank was frequently contaminated (80%) by A. butzleri; our PFGE findings excluded primary contamination of milk, whereas teats and milking machine surfaces could be sources of Arcobacter milk contamination.