Occurrence Of Arcobacter Research Articles

Occurrence and characterization of Arcobacter spp. from ready-to-eat vegetables produced in Southern Italy.

Given that the number of foodborne illness outbreaks linked to the consumption of ready-to-eat vegetables has been widely documented and considering that data on the occurrence of Arcobacter spp. in such foodstuffs are lacking, the aim of the present study was to evaluate the presence of Arcobacter spp. and the occurrence of virulence factors as well as to genotype Arcobacter spp. in ready-to-eat (RTE) vegetable samples, using cultural and biomolecular assays. Arcobacter spp. was detected in 16/110 (14.5%) samples, with A. butzleri being detected in 15/16 and A. cryaerophilus in 1/16 isolates. PCRs aimed at the nine putative virulence genes demonstrated widespread distribution of such genes among A. butzleri and A. cryaerophilus isolates. In addition, multilocus sequence type (MLST) analysis revealed a low genetic diversity within the arcobacters isolates. The results underline the need to develop an appropriate surveillance system based on biomolecular characterization for an integrated microbiological risk assessment of ready-toeat vegetables, and consequently of composite foods.

Detection of Arcobacter spp. in environmental and food samples collected in industrial and artisanal sheep's milk cheese-making plants

The genus Arcobacter include species considered as human foodborne enteropathogens. In recent years, Arcobacter spp. have been frequently isolated from various food of animal origin such as meat, fish and dairy products. However, there is still little information on the occurrence of Arcobacter spp. in the dairy sheep chain. The present study was aimed to assess the prevalence of Arcobacter spp. in the processing environment of 10 industrial and 10 artisanal cheese making plants using sheep milk. A total of 250 samples were collected including food contact and non-food contact surfaces, cheeses, water and wastewater samples. Species identification of isolates was conducted by multiplex PCR and PCR-RFLP. Arcobacter spp. were isolated in 11 out of 20 cheese-making plants (55.0%) with an overall prevalence of 19.2%. Of Arcobacter positive samples 32 (66.6%) were detected from environmental surfaces, 11 (22.9%) from food samples and 5 from wastewater (10.4%). Arcobacter was never recovered from water samples. No differences were observed between industrial and artisanal cheese-making plants. Of 48 isolated strains, 39 (81.3%) were A. butzleri, 5 (10.4%) A. cryaerophilus and 4 (8.3%) A. skirrowii. This is the first investigation reporting the detection of A. cryaerophilus and A. skirrowii from both environmental surfaces and cheeses in dairy plants using sheep milk. Contamination was observed respectively in 6 ricotta (20.0%) and in 3 fresh cheeses (23.1%). The present study indicates the potential risk for human health associated to cheese obtained with pasteurized milk due to cross contamination originating from the processing environment. To minimize the risk of transmission of Arcobacter spp. to consumers through the ingestion of contaminated foods it is essential the implementation of environmental monitoring programs to collect samples from food processing surfaces and to detect the presence of this potential human pathogen.

Occurrence and Antibiotic Resistance of Arcobacter Species Isolates from Poultry in Tunisia

Arcobacter is considered an emergent foodborne enteropathogen. Despite the high prevalence of this genus in poultry, the occurrence of Arcobacter spp. contamination in Tunisia remains unclear. The objectives of this study were (i) to isolate Arcobacter species (A. butzleri and A. cryaerophilus) by the culture method from different species of raw poultry meat, (ii) to verify the isolates by multiplex PCR (m-PCR) assay and matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS), and (iii) to determine the antibiotic resistance profiles of the isolates. A total of 250 poultry product samples (149 chicken and 101 turkey) were collected from various supermarkets in Sfax. The samples consisted of breasts, wings, legs, and neck skins. The overall isolation frequency of Arcobacter spp. was 10.4%. Arcobacter spp. were found in 13.42% of the chicken samples and in 5.49% of the turkey samples. All the acquired isolates were subject to detailed confirmation with subsequent species classification using m-PCR and MALDI-TOF MS. A. butzleri was found in 22 samples (84.61%) and A. cryaerophilus in 4 samples (15.38%). Thus, m-PCR and MALDI-TOF MS were able to detect A. butzleri significantly better than the conventional method (χ2 = 49.1 and P < 0.001). Arcobacter was isolated from poultry in every season, at contamination levels of 30.76, 23.07, 19.23, and 26.92% in summer, spring, autumn, and winter, respectively. The disk diffusion method was used to determine the susceptibility of Arcobacter isolates to six antimicrobial drugs. All A. butzleri isolates (n = 24) were significantly resistant to erythromycin (P = 0.0015), ampicillin (P = 0.001), and ciprofloxacin (P = 0.05). All tested A. cryaerophilus strains (n = 4) were susceptible to ampicillin, gentamicin, and amoxicillin-clavulanic acid. Multidrug resistance was observed in 83% of the Arcobacter spp. isolates. Our study detected Arcobacter spp. in Tunisian poultry; because of their multidrug resistance, these species may constitute a public health problem.

Detection of Arcobacter species in different intestinal compartments of broiler chicken during slaughter and processing.

Arcobacter spp. are commonly present on meat products. However, the source of contamination on chicken meat is under dispute. Since different studies reported contradictory results on the occurrence of Arcobacter spp. inside the intestinal tract of chicken, our study examined four intestinal compartments at four significant production steps during broiler slaughter and processing in the slaughterhouse. Altogether, 157 intestinal tracts from 19 flocks were examined qualitatively and semiquantitatively applying a selective enrichment. Further verification was performed by mPCR and rpoB sequencing. Arcobacter spp. were only detected sporadically in intestinal contents after bleeding (2/32) and in none after scalding (0/32). After defeathering, Arcobacter spp. were detected in 62% (18/29) of the intestinal contents with 28% (8/29) of the duodenal, 21% (6/29) of the jejunal, 3% (1/29) of the cecal, and 55% (16/29) of the colonic samples tested positive with loads up to 24,000 MPN/g in the colonic content. Further 88% (7/8) of colonic tissue samples were tested positive. After evisceration, the prevalences (58/64) and loads of Arcobacter spp. display comparable levels in the intestinal contents like after defeathering. In conclusion, our data point out that Arcobacter spp. are most likely detected in the colonic intestinal compartment of the chicken after defeathering and evisceration. Therefore, not only cross‐contamination originating from the environment inside the slaughterhouse may cause carcass contamination with Arcobacter spp. on broiler chicken carcasses. The detection of Arcobacter spp. in duodenal and jejunal contents as well as in the colonic tissue indicates that there possibly exists an Arcobacter reservoir inside the chicken.

Occurrence ofArcobacter spp. in Brazilian Minas frescal cheese and raw cow milk and its association with microbiological and physicochemical parameters

Some Arcobacter species are considered emerging food-borne pathogens. However, their prevalence in dairy products is not well known. Therefore, the aim of the current research is to know the occurrence of Arcobacter species in Minas frescal cheese and raw cow milk and its potential association with physicochemical and microbiological parameters. Two hundred and one food samples (cheese, n = 98 and milk, n = 103) collected between 2016 and 2017 in São Paulo State, Brazil, were analyzed. Arcobacter was isolated using an enrichment broth with further filtration over Arcobacter agar. Colonies were characterized by ERIC-PCR, m-PCR and PCR-RFLP and, when was necessary, sequencing of 16S rRNA and rpoB genes were also assessed. Additionally, cheese samples were evaluated in relation to total coliform (TC), thermotolerant coliforms (TtC), coagulase positive staphylococci (CoPS) count and moisture. Raw milk samples were assessed for aerobic mesophilic bacteria count, TC, TtC, Dornic acidity, proteins, fats, lactose, defatted dry extract, total solids and somatic cell counts. The occurrence of Arcobacter was 10.2% (10/98) and 16.5% (17/103) in cheese and milk samples, respectively. All isolates were identified as Arcobacter butzleri. On the other hand, Multiple Factorial Analysis showed correlation between the presence of this species and the cheese moisture, while this pathogen was correlated with high counts of mesophilic microorganisms and TC in raw milk. No other evaluated parameter in cheese (TC, CoPS) and milk (TtC, Dornic acidity, proteins, fats, lactose, defatted dry extract, total solids and somatic cell counts) showed significant correlation. To date, this is the first report on A. butzleri in Minas frescal cheese, therefore its presence in dairy products could be considered a potential public health concern.

Occurrence of Arcobacter spp. and correlation with the bacterial indicator of faecal contamination Escherichia coli in bivalve molluscs from the Central Adriatic, Italy

A total of 162 samples of bivalve molluscs (45 mussels and 117 clams) collected between December 2012 and 2014 from harvesting areas of the Central Adriatic were analysed by a culturing method for the presence of Arcobacter spp. Species identification was performed by PCR and sequencing analysis of a fragment of the rpoB gene. Overall, Arcobacter species were detected in 30% of samples, specifically 33% clams and 22% mussels. A. butzleri was the most common species (20% of the samples), followed by A. cryaerophilus (9%) and A. skirrowii (1%). A seasonal association of A. butzleri contamination was detected. A. butzleri was significantly more commonly recovered from samples collected during the winter-spring period (29%) than from those of the summer-autumn (8%). A. cryaerophilus was cultured from 6% to 11% of the samples collected in summer-autumn and winter-spring, respectively, but these differences were not statistically significant. A. skirrowii was recovered from a sample of mussels harvested in May 2014. To identify associations between the occurrence of Arcobacter spp. and E. coli levels, samples were divided into groups generating results with E. coli at >230MPN/100g and E. coli at ≤230MPN/100g, the latter corresponding to EU microbiological criteria allowed for live bivalve molluscs at retail level. A. butzleri was significantly more commonly detected in samples with higher E. coli levels (48%) than in those with lower levels of E. coli (10%), providing evidence for considering E. coli as an index organism for A. butzleri contamination in bivalve molluscs.

Modified isolation method of Arcobacter spp. from different environmental and food samples

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.

Occurrence of Arcobacter in Iranian poultry and slaughterhouse samples implicates contamination by processing equipment and procedures

1. The occurrence of Arcobacter spp. and three pathogenic species of Arcobacter from Iranian poultry carcasses was investigated at different steps of broiler processing to determine critical control points for reducing carcass contamination.2. Samples were collected from (a) cloaca immediately before processing, (b) different points during processing and (c) at different stations in a processing plant of a slaughterhouse in southern Iran.3. After enrichment steps in Arcobacter selective broth, DNA of the samples was extracted and three significant pathogen species of Arcobacter were identified based on polymerase chain reaction (PCR) detection of 16S rRNA and specific species PCR.4. Out of a total of 540 samples, 244 (45%) were positive for Arcobacter spp. Arcobacter butzleri was more frequently detected (73% ± 13.9%) than A. cryaeophilus (9% ± 13.9%) and A. skirrowii (4.1%). In addition, co-colonisation (A. butzleri and A. cryaerophilus) occurred in 13.9% of the positive samples.5. The results indicate a high prevalence of Arcobacter in the investigated slaughterhouse and broiler carcasses and that Arcobacter is not a normal flora of the broilers. Evidence for the presence of Arcobacter in the environment and water of processing plants suggests that these are sources of contamination of poultry carcasses. In addition, contamination of the poultry carcasses can spread between poultry meats in different parts and processes of the slaughterhouse (pre-scalding to after evisceration).

Niche differentiation among mat-forming, sulfide-oxidizing bacteria at cold seeps of the Nile Deep Sea Fan (Eastern Mediterranean Sea)

Sulfidic muds of cold seeps on the Nile Deep Sea Fan (NDSF) are populated by different types of mat-forming sulfide-oxidizing bacteria. The predominant sulfide oxidizers of three different mats were identified by microscopic and phylogenetic analyses as (i) Arcobacter species producing cotton-ball-like sulfur precipitates, (ii) large filamentous sulfur bacteria including Beggiatoa species, and (iii) single, spherical Thiomargarita species. High resolution in situ microprofiles revealed different geochemical settings selecting for the different mat types. Arcobacter mats occurred where oxygen and sulfide overlapped above the seafloor in the bottom water interface. Filamentous sulfide oxidizers were associated with steep gradients of oxygen and sulfide in the sediment. A dense population of Thiomargarita was favored by temporarily changing supplies of oxygen and sulfide in the bottom water. These results indicate that the decisive factors in selecting for different mat-forming bacteria within one deep-sea province are spatial or temporal variations in energy supply. Furthermore, the occurrence of Arcobacter spp.-related 16S rRNA genes in the sediments below all three types of mats, as well as on top of brine lakes of the NDSF, indicates that this group of sulfide oxidizers can switch between different life modes depending on the geobiochemical habitat setting.

Prevalence and Diversity of Arcobacter spp. in the Czech Republic

The aim of this study was to examine 634 samples of chicken, lamb, pork, beef, fish, samples from the intensive animal industry and from poultry for slaughter, as well as from the domestic breeding of poultry, horses, pigs, and lambs, from surface water, and from clinical samples for the presence of Arcobacter. All the samples were examined with a cultivation method, followed by confirmation by multiplex PCR. The method of multiplex PCR applied directly to a liquid medium after enrichment was applied only to the samples with the highest probability of the presence of arcobacters. Arcobacter spp. were detected in 11.8% of the samples, of which A. butzleri, A. cryaerophilus, and A. skirrowii were found in 6.6, 5.1, and 0.2% of the samples, respectively. The sources of the arcobacters were chicken meat from the retail market, intensive animal production facilities, domestic chicken breeding facilities, lamb raising environments, surface water and wastewater, and beef swabs taken in a meat processing factory. No occurrence of arcobacters was identified in the swabs from slaughter turkeys, ducks, and wild poultry. No arcobacters were found in horse and pig breeding environments, on pork, or on the swabs of fish. Forty-two rectal swabs taken from humans were also free of Arcobacter. Seventeen isolates of Arcobacter were further identified by sequencing the 16S rRNA gene. Varied genotypes were observed among A. butzleri from chicken meat and chicken breeds, and A. cryaerophilus from wastewater and chicken breeds. They were similar to the genotypes present in wastewater, porcine feces, human stool, and human blood obtained from databases. Our results revealed that the chicken meat from the retail market is an important source of arcobacters. Cross-contamination during handling of chicken carcass practices could play a key role in the spread of Arcobacter.

Characterization of the Arcobacter contamination on Belgian pork carcasses and raw retail pork

In the present study, the occurrence of Arcobacter was assessed at four sites on 169 porcine carcasses (foreleg, chest, pelvis and ham) at different stages of slaughter and 47 pork products at retail. Carcass swab samples were enriched in Arcobacter broth containing 5-fluorouracil, amphotericine B, cefoperazone, novobiocine and trimethoprim as selective supplement. After microaerobic incubation, arcobacters were isolated using Arcobacter selective agar plates, containing the selective supplement described above. Some carcass samples and all pork samples were also examined quantitatively. All 862 isolates were identified by a species-specific m-PCR-assay and 182 isolates were further characterized by ERIC-PCR. Arcobacters were isolated from one or more sampling places on 96.4% of the carcasses, with the foreleg and the chest area as the two most contaminated sites. Furthermore, A. cryaerophilus was the most common species. Chilling decreased the number of positive carcasses, but did not eliminate all arcobacters. Direct isolation revealed that only a few carcasses were contaminated with arcobacters on foreleg and/or chest at levels higher than 10 2 cfu/100 cm 2. Characterization demonstrated a large heterogeneity among the isolates, with ten genotypes present on more then one site per carcass. Fourteen genotypes were simultaneously present on carcasses from different herds slaughtered on the same day, which may indicate cross-contamination. Arcobacters were present in 21% of the pork samples taken at retail, but contamination levels did not exceed 100 cfu per gram. Characterization of the A. butzleri and A. cryaerophilus isolates indicated an additional contamination during processing at retail.

Discrepancy Between the Occurrence of Arcobacter in Chickens and Broiler Carcass Contamination

Both Campylobacter and Arcobacter are commonly present on broiler carcasses. For Campylobacter, the superficial contamination originates predominantly from fecal contamination during slaughter. In contrast with Campylobacter, the source of the Arcobacter contamination is not clear. In several studies, arcobacters have been isolated in poultry processing plants from the carcasses and slaughter equipment, but not from the intestinal content. In literature, contradictory reports about the Arcobacter colonization of the chicken gut have been published. In most of those studies, arcobacters were not isolated from cecal content nor from litter or the feathers, though some studies reported the isolation of arcobacters from cloacal swab samples. The present study assessed if arcobacters are part of the chicken intestine, skin, or feather flora. Because no isolation protocol has been validated for poultry intestinal content, a previously developed Arcobacter isolation procedure for feces from livestock animals was first validated. With this method, a good repeatability, in-lab reproducibility and sensitivity, and a good suppression of the chicken fecal accompanying flora were achieved when 125 mg/L of 5-fluorouracil, 10 mg/L of amphotericine B, 100 mg/L of cycloheximide, 16 mg/L of cefoperazone, 64 mg/L of novobiocine, and 64 mg/L of trimethoprim were applied. The validated method was used to examine the presence of arcobacters in and on living chickens of 4 flocks at slaughter age. Because arcobacters were not isolated from the intestinal tract nor from the skin or feathers of the birds, this study was not able to identify arcobacters as part of the intestinal or skin flora, nor could confirm the role of process water as reservoir. However, the results clearly demonstrated that the time period for processing the samples and the way of sample collection are crucial in the interpretation of epidemiological studies. As the reservoir of the carcass contamination remains unidentified, studies about the capacity of arcobacters to colonize the chicken intestinal tract may contribute in the assessment of the transmission routes of this emerging foodborn pathogen.

Isolation of Arcobacter spp from poultry carcasses, in Brazil

Fourty eight isolates of Arcobacter spp were obtained from 37 poultry carcasses, from abattoir, among 80 carcasses examined. Attempts for culturing were made from the skin and muscle, resulting on 25 positive cultures from muscle and 23 from skin. Classification was achieved by phenotypic characterization and PCR and multiplex PCR, resulting 41 samples of Arcobacter butzleri and 07 Arcobacter sp. This is the first report on the occurrence of Arcobacter in animal carcasses in Brazil.