Presence Of Arcobacter Research Articles

Antibiotic-Resistant Arcobacter spp. in commercial and smallholder farm animals in Asante Akim North Municipality, Ghana and Korogwe Town Council, Tanzania: a cross-sectional study

BackgroundArcobacter species are considered emerging foodborne pathogens that can potentially cause serious infections in animals and humans. This cross-sectional study determined the frequency of potentially pathogenic Arcobacter spp. in both commercial and smallholder farm animals in Ghana and Tanzania. A total of 1585 and 1047 (poultry and livestock) samples were collected in Ghana and Tanzania, respectively. Selective enrichment media, along with oxidase and Gram testing, were employed for isolation of suspected Arcobacter spp. and confirmation was done using MALDI-TOF MS. Antibiotic susceptibility was assessed through disk diffusion method and ECOFFs were generated, for interpretation, based on resulting inhibition zone diameters.ResultsThe overall Arcobacter frequency was higher in Ghana (7.0%, n = 111) than in Tanzania (2.0%, n = 21). The frequency of Arcobacter in commercial farms in Ghana was 10.3% (n/N = 83/805), while in Tanzania, it was 2.8% (n/N = 12/430). Arcobacter was detected in only 3.6% (n/N = 28/780) of the samples from smallholder farms in Ghana and 1.5% (n/N = 9/617) of the samples from Tanzania. For commercial farms, in Ghana, the presence of Arcobacter was more abundant in pigs (45.1%, n/N = 37/82), followed by ducks (38.5%, n/N = 10/26) and quails (35.7%, n/N = 10/28). According to MALDI-TOF-based species identification, Arcobacter butzleri (91.6%, n/N = 121/132), Arcobacter lanthieri (6.1%, n/N = 8/132), and Arcobacter cryaerophilus (2.3%, n/N = 3/132) were the only three Arcobacter species detected at both study sites. Almost all of the Arcobacter from Ghana (98.2%, n/N = 109/111) were isolated during the rainy season. The inhibition zone diameters recorded for penicillin, ampicillin, and chloramphenicol allowed no determination of an epidemiological cut-off value. However, the results indicated a general resistance to these three antimicrobials. Multidrug resistance was noted in 57.1% (n/N = 12/21) of the Arcobacter isolates from Tanzania and 45.0% (n/N = 50/111) of those from Ghana. The type of farm (commercial or smallholder) and source of the sample (poultry or livestock) were found to be associated with multi-drug resistance.ConclusionsThe high levels of MDR Arcobacter detected from farms in both countries call for urgent attention and comprehensive strategies to mitigate the spread of antimicrobial resistance in these pathogens.

Survival capacity of Arcobacter butzleri at different incubation temperatures in drinking water

Arcobacter represents a zoonotic emerging pathogen with increasing importance for public health and drinking water has been cited as a major risk factor for its dissemination. The aim of this work was to evaluate the survival capacity of Arcobacter in different water matrixes stored at different temperatures. Three different water matrixes were used, including potable water with a chlorine concentration of 0,5 mg/mL, non-chlorinated water and non-chlorinated water added with an 11% of organic matter. Each matrix was inoculated in a 1/10 proportion with 103 and 105 Arcobacter pools, divided into 4 different subsamples, in order to be incubated at 0°C, 5°C, 12 °C and 25°C by up to 15 days. The presence of Arcobacter in each matrix was determined on days 1, 3, 5, 7, 9, 11, 13 and 15. Results obtained show that this bacterium can survive in all the water matrixes evaluated, regardless of the presence or not of residual disinfecting agent. Also, the amount of CFU/mL inoculated in water correlates with the number of bacteria that can survive on it, and that incubation temperature has a significant effect over the bacterial survival.

A Review on the Prevalence of Arcobacter in Aquatic Environments

Arcobacter is an emerging pathogen that is associated with human and animal diseases. Since its first introduction in 1991, 33 Arcobacter species have been identified. Studies have reported that with the presence of Arcobacter in environmental water bodies, animals, and humans, a possibility of its transmission via water and food makes it a potential waterborne and foodborne pathogen. Therefore, this review article focuses on the general characteristics of Arcobacter, including its pathogenicity, antimicrobial resistance, methods of detection by cultivation and molecular techniques, and its presence in water, fecal samples, and animal products worldwide. These detection methods include conventional culture methods, and rapid and accurate Arcobacter identification at the species level, using quantitative polymerase chain reaction (qPCR) and multiplex PCR. Arcobacter has been identified worldwide from feces of various hosts, such as humans, cattle, pigs, sheep, horses, dogs, poultry, and swine, and also from meat, dairy products, carcasses, buccal cavity, and cloacal swabs. Furthermore, Arcobacter has been detected in groundwater, river water, wastewater (influent and effluent), canals, treated drinking water, spring water, and seawater. Hence, we propose that understanding the prevalence of Arcobacter in environmental water and fecal-source samples and its infection of humans and animals will contribute to a better strategy to control and prevent the survival and growth of the bacteria.

Microbial diversity in water and animal faeces: a metagenomic analysis to assess public health risk

ABSTRACT Environmental DNA (eDNA) offers a new avenue for investigating changes in the water microbial community associated with faecal contamination. Faeces in drinking water might include pathogens, which result in serious waterborne diseases in humans. Therefore, drinking water requires comprehensive information about microbial diversity that comes from faecal contamination of different sources to reduce the risk of gastrointestinal diseases. Here, we investigated the microbial diversity of water and faecal samples at 15 recreational campgrounds in New Zealand. In total, 42 faecal (two rabbits, seven ducks, seven ruminants, seven passerines, nine possum and ten Pukeko) and 75 water (37 intakes and 38 taps) samples were analysed using 16S rRNA metabarcoding. Our results suggested that water samples harbour a higher microbial diversity than faeces. Canonical correspondence analysis of bacterial communities and NeighborNet tree of recognised pathogens showed clustering of samples from similar sources. Phylogenetic analyses showed evidence for the presence of Arcobacter and Sulfurospirillum and indicator organisms Escherichia and enterococci in water, while Campylobacter was mainly found in faeces. These findings provide novel insights toward understanding the quality of drinking water and allow future use for the identification of faecal contamination in water.

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.

Identification of 16S rRNA and Virulence-Associated Genes of Arcobacter in Water Samples in the Kathmandu Valley, Nepal.

This study aimed to determine the prevalence of Arcobacter and five associated virulence genes (cadF, ciaB, mviN, pldA, and tlyA) in water samples in the Kathmandu Valley, Nepal. A total of 286 samples were collected from deep tube wells (n = 30), rivers (n = 14), a pond (n = 1), shallow dug wells (n = 166), shallow tube wells (n = 33), springs (n = 21), and stone spouts (n = 21) in February and March (dry season) and August (wet season), 2016. Bacterial DNA was extracted from the water samples and subjected to SYBR Green-based quantitative PCR for 16S rRNA and virulence genes of Arcobacter. The 16S rRNA gene of Arcobacter was detected in 36% (40/112) of samples collected in the dry season, at concentrations ranging from 5.7 to 10.2 log copies/100 mL, and 34% (59/174) of samples collected in the wet season, at concentrations of 5.4–10.8 log copies/100 mL. No significant difference in Arcobacter 16S rRNA gene-positive results was observed between samples collected in the two seasons (p > 0.05). Seventeen (17%), 84 (84%), 19 (19%), 23 (23%), and 17 (17%) of the 99 Arcobacter 16S rRNA gene-positive samples were also positive for cadF, ciaB, mviN, pldA, and tlyA, respectively. At least one virulence gene was detected in 87 (88%) of the 99 Arcobacter 16S rRNA gene-positive samples. The presence of Arcobacter and the virulence genes in these samples illustrates the persistence of pathogenic bacteria in the environment and highlights the importance of regular monitoring of water for pathogens.

The Use of a DNA-Intercalating Dye for Quantitative Detection of Viable Arcobacter spp. Cells (v-qPCR) in Shellfish.

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.

Detection of Arcobacter spp. in food products collected from Sicilia region: A preliminary study.

The aim of the study was to evaluate the occurrence of Arcobacter spp. in food samples collected from Sicilia region. A total of 91 food products of animal origin (41 meat, 17 fresh milk, 18 shellfish) and 15 samples of fresh vegetables, were examined by cultural method and confirmed by biochemical analysis and PCR methods. The detection of Arcobacter spp. was performed, after selective enrichment, on two selective agar plates: Arcobacter agar and mCCD (modified charcoal cefoperazone deoxycholate) agar supplemented with CAT (Cefoperazone, Amphotericin B and Teicoplanin). Arcobacter species were isolated using the membrane filtration technique. In 13 (14.3%) out of the 91 tested samples, the presence of Arcobacter spp. was found: the isolates were confirmed by multiplex PCR and identified as belonging to the species A. butzleri and A. cryaerophilus. The highest prevalence rate was observed in chicken meat (8.8%) followed by shellfish (3.3%). Negative results have been obtained for raw milks and vegetables samples. The preliminary study highlights the importance of this emerging pathogen and the need for further studies on its prevalence and distribution in different types of food for human consumption.

Isolation and determination of antimicrobial resistance of Arcobacter species isolated from animal faeces in the Diyarbakir region of Turkey using the 16S rDNA-RFLP method

In this study, the presence of Arcobacter spp. was investigated in the faeces of cattle, sheep, goats, dogs and cloacal swab samples of chickens using the 16S rDNA-RFLP method. The prevalence of Arcobacter in these species was found to be 13%, 12%, 16%, 4% and 33%, respectively. On the other hand, Arcobacter spp. could not be isolated from rabbit faeces. A total of 78 (13%) Arcobacter spp. isolates were obtained from the 500 faecal samples and 100 cloacal swab samples examined in this study. From these 78 Arcobacter isolates, 24 (30.8%), 20 (25.6%), 11 (14.1%), 8 (10.7%), 4 (5.1%), 3 (3.9%) and 2 (2.6%) were identified by 16S rDNA-RLFP as A. cryaerophilus, A. butz- leri, A. skirrowii, A. cloacae, A. cibarius, A. halophilus, and A. nitrofigilis, respectively. All A. cryaerophilus (n = 24) isolates were found to be resistant to cloxacillin; all A. butzleri (n = 20) and A. skirrowii isolates were found to be resistant to penicillin/novobiocin, cefoperazone, tetracycline and cloxacillin. It was determined in this study that clinically healthy cattle, sheep, goats, dogs and chickens are reservoirs of Arcobacter spp.

Isolation, molecular identification and quinolone-susceptibility testing of Arcobacter spp. isolated from fresh vegetables in Spain

Some species of the Arcobacter genus are considered emerging foodborne and waterborne enteropathogens. However, the presence of Arcobacter spp. in vegetables very little is known, because most studies have focused on foods of animal origin. On the other hand, quinolones are considered as first-line drugs for the treatment of infection by campylobacteria in human patients, but few data are currently available about the resistance levels to these antibiotics among Arcobacter species. Therefore, the aim of this study was to investigate the presence and diversity of arcobacters isolated from fresh vegetables such as lettuces, spinaches, chards and cabbages. Resistance to quinolones of the isolates was also investigated.One hundred fresh vegetables samples purchased from seven local retail markets in Valencia (Spain) during eight months were analysed. The study included 41 lettuces, 21 spinaches, 34 chards and 4 cabbages. Samples were analysed by culture and by molecular methods before and after enrichment. By culture, 17 out of 100 analysed samples were Arcobacter positive and twenty-five isolates were obtained from them. Direct detection by PCR was low, with only 4% Arcobacter spp. positive samples. This percentage increased considerably, up 20%, after 48 h enrichment. By polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP), 17 out of the 25 isolates were identified as A. butzleri and 8 as A. cryaerophilus. Only two A. butzleri isolates showed resistance to levofloxacin and ciprofloxacin. The sequencing of a fragment of the QRDR region of the gyrA gene from the quinolones-resistant isolates revealed the presence of a mutation in position 254 of this gene (C-T transition).This study is the first report about the presence of pathogenic species of Arcobacter spp. in chards and cabbages and confirms that fresh vegetables can act as transmission vehicle to humans. Moreover, the presence of A. butzleri quinolone resistant in vegetables could pose a potential public health risk.

Incidence of Arcobacter spp. in fresh seafood from retail markets in Mumbai, India

The occurrence of Campylobacter-like bacteria in fresh fish sold in retail markets of Mumbai was studied. Microaerophilic bacteria isolated following enrichment in Bolton broth and selective plating on Preston blood agar were subjected to biochemical identification followed by polymerase chain reaction to detect Campylobacter, Arcobacter and Helicobacter genera. Arcobacter spp. was isolated from 10 (25 %) of the 40 seafood samples analysed. Based on the comparison of partial 16S rDNA gene sequence, the Arcobacter isolates of this study were identified as Arcobacter butzleri, A. skirrowii and A. cibarius, with A. butzleri being the predominant species. The presence of arcobacter represents a health risk and requires further study to determine the sources of contamination of seafood and the pathogenic potential of Arcobacter spp.

First detection of Arcobacter sp. in Eurasian collared doves (Streptopelia decaocto).

Data about the presence of Arcobacter in wild birds are currently lacking. In this study cloacal swabs from 95 collared doves (Streptopelia decaocto), submitted to the Department of Veterinary Medical Sciences (Bologna, Italy) between 2011 and 2013 from various urban and suburban areas of the Emilia-Romagna region (Northern Italy), were tested for the presence of Arcobacter sp. by a rRNA 23S nested Polymerase Chain Reaction (PCR). Eighteen out of 95 (19%) samples showed the expected PCR product. Further cultural and molecular studies are needed to assess the Arcobacter prevalence in wild birds and elucidate their potential epidemiological role as source of animal and human infections.

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).

A Comparison of Media for the Isolation of Arcobacter spp. from Retail Packs of Beef

In order to determine the most effective protocol for the isolation of Arcobacter spp. from retail packs of beef, three published methods (A, B, and C) were selected. In addition, a modified version of method B was studied (method D). The ability of the four methods to isolate Arcobacter from standardized beef samples (n = 80) was compared with presumptive Arcobacter isolates being identified to genus and species level, using multiplex PCR methods. The presence of Arcobacter in enrichment broths was also investigated using PCR techniques. Overall, the modified enrichment and selection media of Johnson and Murano (method D) gave the highest recovery of Arcobacter. Recovery using these media was enhanced by incubating the enrichment and selection media in a microaerobic cabinet rather than air, and the inclusion of streaking the enrichment broth onto selective agar after 24 h in addition to 48 h. Method D yielded significantly more Arcobacter-positive samples of beef (P < 0.01) than did the three other methods investigated.

Isolation of various Arcobacter species from domestic geese ( Anser anser)

In this study, the prevalence and distribution of various Arcobacter spp. were investigated in samples taken from the cloacae of healthy domestic geese raised in Turkey. A membrane filtration technique with a non-selective blood agar was employed after enrichment in Arcobacter enrichment broth (AEB) to isolate a wide range of Arcobacter spp. In addition, the isolates were characterized phenotypically and identified at species level using a multiplex-PCR assay. A total of 90 cloacal swab samples taken from geese, collected on three farms (18, 25, 47 samples, respectively), were examined. Of the samples examined, 16 (18%) were found positive for Arcobacter. One Arcobacter species was isolated from each bird. Of the 16 Arcobacter isolates, 7 (44%), 7 (44%) and 2 (12.5%) were identified by m-PCR as A. cryaerophilus, A. skirrowii and A. butzleri, respectively. The present study indicates that domestic geese can harbour a variety of Arcobacter spp. in their cloacae. The presence of Arcobacter in geese may be of significance as reservoirs in their dissemination. Detailed research is needed for better understanding of the epidemiology and zoonotic potential of this emerging pathogen.

Detection and diversity of various Arcobacter species in Danish poultry

The prevalence and diversity of different Arcobacter spp. in various poultry species in Denmark were investigated using cultural and multiplex PCR methods. A pool of three fresh droppings obtained at the production site from 70 broiler chicken flocks aged 4–5 weeks was examined. In addition, pools of 10 cloacal swabs taken at the abattoir prior to stunning from each of 15, and 37 duck and turkey flocks, respectively, were analyzed. Thirty fresh broiler chicken carcasses and 29 cloacal swabs from the respective viscera were also examined at the abattoir. Finally, 10 caecal and 10 cloacal swabs from ducks at the abattoir were analyzed individually. In total, 85 Arcobacter isolates were obtained. Of these 45, 20 and 7 were identified as Arcobacter butzleri, Arcobacter cryaerophilus and Arcobacter skirrowii, respectively, using a multiplex PCR. Interestingly, some chicken isolates of A. butzleri showed urease activity, and 6 out of seven A. skirrowi isolates were unable to hydrolyse indoxyl acetate. All chicken carcasses examined were found positive for A. butzleri and/or A. cryaerophilus, whereas 21 (72%) of the 29 chicken cloacal swabs were positive for either A. butzleri (13) or A. cryaerophilus (9). Three (4.3%) out of 70 chicken flocks analyzed were positive only for A. cryaerophilus. Of the ten ducks examined individually, 7 carried A. skirrowii and/or A. cryaerophilus in their cloacae. None of the respective caecal samples were positive. Of the remaining 15 duck flocks, 11 (73%) were positive for A. cryaerophilus (7), A. butzleri (2) or A. skirrowii (2). Four (11%) of the 37 turkey flocks analyzed harboured either A. butzleri or A. cryaerophilus. The carriage rate of Arcobacter was higher in live ducks than those of live broiler chickens and turkeys in the present study. In addition, chicken carcasses slaughtered in Denmark were found to be contaminated with Arcobacter. The presence of Arcobacter spp. both on chicken carcasses and in poultry intestine may be of significance to human health.