Typing of Yersinia Enterocolitica Isolates by ITS Profiling,REP‐ and ERIC‐PCR

Thirty‐five Yersinia enterocolitica strains isolated from humans, pigs and foxes were analysed by genotyping including intergenic transcribed sequence (ITS) profiling, REP‐ and ERIC‐PCR. ERIC‐PCR revealed the presence of seven different genotypes. Amplification of the 16S‐23S rDNA spacer region by ITS profiling gave similar results with nine different genotypes. REP‐PCR was found to be more discriminatory for typing of Y. enterocolitica than ERIC‐PCR and ITS profiling. Fifteen different DNA patterns were obtained by this technique. Based on data obtained by three methods it was found that: (i) Y. enterocolitica strains belonging to the same serotype can represent different genotypes and vice versa; (ii) isolates recovered from humans, pigs and foxes exhibit limited heterogeneity and, independent of the origin, one or two prevailing genotypes were always observed; and (iii) many human Y. enterocolitica isolates shared common genotypes with porcine isolates.     

The anatomical distribution and antimicrobial susceptibility of yeast species isolated from healthy dogs

The aim of this work was to identify the predominant yeast species present at different anatomical sites in healthy dogs and to determine their in vitro antimicrobial susceptibility using a broth microdilution assay. Samples were collected from the preputial, vaginal, oral and perianal mucosae and the isolates cultured were identified according to their morphological characteristics and biochemical profile. Malassezia pachydermatis was the most commonly isolated yeast, followed by Candida parapsilosis, Candida tropicalis, Candida albicans, Saccharomyces cerevisiae and Rhodotorula spp.Minimum inhibitory concentrations of the azole derivatives ketoconazole, itraconazole and fluconazole against Candida spp. were 0.03–16 μg/mL, 0.06 to >16 μg/mL and 0.5–64 μg/mL, respectively and Candida isolates were sensitive to caspofungin and amphotericin B. Although all isolates of M. pachydermatis were sensitive to itraconazole, fluconazole, ketoconazole and amphotericin B, they were found to be resistant to caspofungin. The study has highlighted that Candida spp., M. pachydermatis, S. cerevisiae and Rhodotorula spp. are part of the normal canine surface microbiota and some of these organisms exhibit in vitro resistance to commonly used antimicrobials.     

Isolation of antifungal-resistant yeasts from bulk milk in Japan

We isolated ascomycetous yeasts including Candida species, that originally belonged to the genus Candida, from bulk milk in the Aichi area of Japan, and determined the minimum inhibitory concentrations (MICs) of antifungal drugs on these isolates by conducting E-tests. We isolated 7 human pathogenic species (14 isolates) from 14 bulk milk samples: 5 Candida species of yeasts, and 2 Candida-related species. Two isolates of C. albicans and C. inconspicua were resistant to fluconazole (MIC >32 mg/l). One isolate of C. krusei was resistant to both azoles (fluconazole: >256 mg/ml and itraconazole: 4 mg/l). One isolate of C. catenulata might be resistant to amphotericin B (>32 mg/l).     

Comparison of ITS Profiling,REP‐ and ERIC‐PCR of Salmonella Enteritidis Isolates from Poland

Thirty‐one Salmonella Enteritidis strains isolated from chickens, broilers and hens were analysed by genotypic typing including REP‐PCR, ERIC‐PCR and ITS profiling (PCR‐ribotyping). Analysis of DNA banding patterns generated by REP‐PCR revealed the presence of 22 different genotypes, which were grouped by dendrogram analysis into three distinct lineages (maximum similarity approx. 50%). Each isolate of S. Enteritidis analysed by ERIC‐PCR generated an individual DNA pattern. Again, these isolates could be divided into three distinct genomic groups (maximum similarity approx. 60%) by their ERIC‐PCR fingerprints. REP‐ and ERIC‐PCR were found to be more discriminatory for typing of S. Enteritidis than ITS profiling. Amplification of the 16S‐23S rDNA spacer region gave nine different profiles of DNA, subdivided into two closely related groups by dendrogram analysis. In summary, data obtained by genotyping methods for S. Enteritidis isolates from regions located in the south‐west and the central parts of Poland revealed an enormous heterogeneity among analysed samples, and proved that REP‐ and ERIC‐PCR are highly discriminatory techniques, which can be used, in addition to conventional methods, in epidemiological studies of S. Enteritidis infections.     

Prevalence,Molecular Characterization and Antimicrobial Resistance of Thermophilic <Emphasis Type="Italic">Campylobacter</Emphasis> Isolates from Cattle,Hens, Broilers and Broiler Meat in South-eastern Italy

Eleven cattle farms, 8 layer farms, 7 broiler farms and 30 broiler meat samples were investigated in south-eastern Italy throughout 2003 to evaluate the prevalence, the molecular type and antimicrobial resistance of thermophilic Campylobacters. A total of 398 samples were analysed. One Campylobacter isolate for each positive faecal swab and three isolates per positive broiler meat sample were selected for further analysis. Multiplex PCR was performed for species-level identification and PCR-RFLP of the flagellin A gene for genotyping. Resistance to 14 antimicrobials was studied in 188 Campylobacter isolates. Prevalence of campylobacters was high both on farms (100%) and in food samples (73%). On 4/11 cattle farms and on 10/15 poultry farms more than one species was isolated. The presence of more than one genotype was found on 8/11 cattle farms, on 10/15 poultry farms and in 8/22 Campylobacter-positive food samples. High rates of resistance to quinolone were observed: 9/31 (29%) C. jejuni bovine isolates, 4/22 (18%) C. jejuni poultry isolates, and 14/26 (54%) C. coli poultry isolates. Resistance to sulphamethoxazole-trimethoprim was also observed frequently: 18/26 (69%) of the avian C. coli strains, 25/31 (80%) of the C. jejuni strains isolated from poultry and 15/22 (68%) of those isolated from cattle were resistant. There was a significant difference between the rate of resistance to macrolides of C. coli and C. jejuni isolated in poultry, which amounted to 23% and 3%, respectively. This study provided data on the prevalence and antimicrobial resistance of thermophilic campylobacters in south-eastern Italy and confirmed that flaA-typing is an efficient tool to study the epidemiology of Campylobacter strains in short-term investigations.     

Detection of PRRSV circulation in herds without clinical signs of PRRS: comparison of five age groups to assess the preferred age group and sample size

The extraordinary ability of Cryptococcus species to cause disease has focused the attention of scientists on finding ways to improve their identification methods. In this study, PCR-REA, manual methods (morphological and biochemical characteristics), API 20C and VITEK 2 were used to test identify a total of 30 Cryptococcus spp. from human and veterinary sources. PCR-REA was performed using the capsular region as amplification target followed by restriction with the enzymes AgeI, BsmFI and HpaII. PCR-REA identified the strains as C. neoformans var. grubii (n=19) and C. gattii (n=8). There was no significant difference between the API 20C AUX and VITEK 2 when compared to manual methods for the identification of Cryptococcus spp. However, none of these non-manual methods were able to detect C. gattii samples. PCR-REA showed a greater level of concordance with the manual method, besides being faster and more sensitive than the other methods. Therefore, it is indicated for routine identification of Cryptococcus spp. strains.     

动物源性沙门氏菌的耐药性分析及氟苯尼考类耐药基因的鉴定

为了解动物沙门氏菌的流行情况和药物敏感性及氟苯尼考耐药株的耐药基因分布,本试验对临床上疑似患沙门氏菌病的病料进行病原分离和细菌的多重PCR鉴定;采用K-B法测定分离株对23种抗菌药物的敏感性;选择氟苯尼考耐药菌株扩增floR、fexA、fexB、cfr和pexA基因。结果显示,共鉴定出61株沙门氏菌,其中肠炎沙门氏菌10株,鸡白痢沙门氏菌12株,鼠伤寒沙门氏菌39株。所有菌株对青霉素、红霉素、万古霉素耐药,90.16%对6种及6种以上抗菌药耐药。floR基因广泛存在于鼠伤寒沙门氏菌氟苯尼考耐药菌株中(8/12,66.67%),未发现其他耐药基因。研究结果表明鼠伤寒沙门氏菌是鹅源分离株中的优势血清型;floR基因主要介导沙门氏菌对氟苯尼考耐药性,但可能还存在其他机制。     

Phenotypic and genotypic characteristics of Mycobacterium isolates from fighting fish Betta spp. in Malaysia

Mycobacteriosis due to mycobacteria is one of the most common bacterial diseases in ornamental fish. We describe here the phenotypic and genotypic characteristics of Mycobacterium isolates from fighting fish Betta spp. using ATCC Mycobacterium marinum, Mycobacterium fortuitum and Mycobacterium chelonae as references. A total of four isolates (M1, M2, M3, M4) were obtained from four out of 106 fish samples using selective agar, and identified to Mycobacterium genus using acid-fast staining and 16s rRNA gene-based genus specific polymerase chain reaction. DNA sequencing and NCBI-BLAST analysis further identified isolate M1 as M. marinum and isolates M2, M3, M4 as M. fortuitum. Morphological, physiological and biochemical tests were carried out for phenotypic characterizations. Universal M13 and wild-type phage M13 RAPD dendogram was generated to illustrate the genetic relationship of the isolates and reference strains.     

First isolation and identification of Elizabethkingia meningoseptica from cultured tiger frog, Rana tigerina rugulosa

Elizabethkingia meningoseptica has been recognised as an occasional but serious opportunistic bacterial pathogen to human beings. Recently, it was frequently isolated from tiger frog, Rana tigerina rugulosa, with cataract disease, which is the most common disease of unknown aetiology of frogs in Hainan, China. The purpose of this study was to identify and characterise the bacterial strains isolated from the recent outbreaks of cataract disease in farmed tiger frog in Hainan, China, and to evaluate their pathogenicity to the frog and their sensitivity to 20 chemotherapeutic agents.The 16S rRNA gene sequences of strains W0701 (1478 bp), W0702 (1477 bp) and W0703 (1478 bp) showed 98.6–98.7% similarity with the sequence of E. meningoseptica type strain (ATCC 13253) and 99.9–100% similarity with that of E. meningoseptica NTU 870424-IL. Six strains (W0701–W0706) were selected to represent 24 isolates retrieved from six moribund frogs. The morphological, physiological and biochemical characteristics of the six representative isolates were consistent with those of E. meningoseptica strains. The organisms were only susceptible to vancomycin and moderately susceptible to cefoperazone among the 20 investigated chemotherapeutic agents. Virulence test with strain W0702 was conducted and pathogenicity (by intramuscular injection) was demonstrated in the tiger frog. In conclusion, 24 isolates obtained from frogs with cataract disease were the E. meningoseptica strains highly pathogenic to tiger frog, and this is the first report of E. meningoseptica as a pathogen for tiger frog.     

Molecular Diversity of Campylobacter coli and C. jejuni Isolated from Pigs at Slaughter by flaA‐RFLP Analysis and Ribotyping

A population of porcine isolates of Camplobacter jejuni (n=11) and C. coli (n=17) were examined for genotypic relatedness employing ribotyping, as well as polymerase chain reaction–restriction fragment length polymorphism (PCR–RFLP) analysis of the flagellin (fla)A gene locus. PCR was employed to amplify a 533 bp fragment from the flaA gene, including the previously described short variable region (SVR), employing the novel primers, A2 and A1 and successfully generated this amplicon for all wild‐type strains examined (n=28) of both C. jejuni and C. coli, as well as with both type strains, i.e. C. jejuni NCTC 11351 and C. coli NCTC 11366. Individual genotypes were assigned to each isolate typed employing the four typing methods (flaA‐RFLP_{Hae III}, flaA‐RFLP_{Pst I} ribotyping_{Hae III} and ribotyping_{Pst I},) and were assigned an arbitrary genotype code in ascending alphabetical order in comparison with a database of established genotypes for each of the methods employed. This study showed that several flaA‐RFLP and ribopatterns existed within C. jejuni and C. coli, and demonstrated a heterogeneous diversity of strains occurring in the pigs examined. Ribotyping of strains with 16S and 23S rRNA with Pst I and Hae III digested chromosomal DNA allowed subdivision of strains into nine and eight groups, respectively. RFLP analyses with Pst I and Hae III digests probed with the flaA gene probe allowed subdivision of strains into eight and eleven subtypes, respectively. Employment of RFLP with the flaA nucleic acid probe and Hae III digests produced the greatest amount of variation of any genotyping scheme employed. Although there was a high degree of variability demonstrated by both typing methods, most isolates (>60%) clustered into four main genotypes, i.e. genotypes A–D. FlaA‐PCR–RFLP typing demonstrated that the majority of isolates, 67.9 and 60.7%, were included in these four main genotypes for Pst I and Hae III restriction digests, respectively, although there was a high prevalence (7/11; 63.6%) of fla_{Hae III} genotype A occurring within the C. jejuni isolates. Likewise, ribotyping studies demonstrated that most isolates were clustered into these four main genotypes, accounting for 81.5 and 60.7% of isolates for Pst I and Hae III restriction digests, respectively. This may indicate that the clonal population of campylobacters within this pig population is largely composed of persistent and dominant types, with a smaller number of hypervariable subtypes. Such data may useful in determining epidemiological routes of transmission of campylobacters from animal to animal, as well as helping to identify virulence determinants in persistent subtype populations.     

四川省部分地区山羊沙门氏菌健康带菌率及耐药性调查

为了解四川省山羊沙门氏菌带菌情况,本试验采集了四川省部分地区5个规模化山羊养殖场表观健康山羊粪便196份,经BPW预增菌、TTB选择性增菌后,采用靶向invA基因的PCR方法检测沙门氏菌的带菌率;对阳性样本进行细菌分离鉴定,随机选择25株沙门氏菌分离株测定其对15种抗菌药物的敏感性。结果显示,山羊沙门氏菌的平均带菌率为54.59%。药敏试验结果显示,分离株均对丁胺卡那敏感,而对其余14种抗菌药物呈不同程度的耐药,多重耐药菌株占88%,其中耐2~7种药物的占52%,耐9~13种药物的占36%。结果表明,四川省部分地区山羊的沙门氏菌健康带菌率较高,且多重耐药性普遍,其公共卫生意义值得进一步研究。     

Characterization of integrons in multiple antimicrobial resistant Escherichia coli isolates from bovine endometritis

To assess the prevalence of antimicrobial resistance and three classes of integrons in Escherichia coli (E. coli) strains (n = 57) isolated from bovine endometritis in Inner Mongolia of China, antimicrobial susceptibility and the presence of three types of integrons were characterized. Most isolates were susceptible to ceftiofur, furazolidone, ciprofloxacin and enrofloxacin, while 57 isolates were all resistant to sulfamethoxydiazine and trimethoprim. High resistant incidence rates were exhibited to sulfadiazine, tetracycline, oxytetracycline, cefazolin, chloramphenicol. Forty-six of 57 E. coli strains were resistant to above 10 antibiotics (80.70%). The integrase gene and gene cassettes of integrons were amplified by PCR. DNA sequencing and analysis were used to identify the genetic content of the integron-variable regions. Neither class II nor class III integron was detected, while 36.8% (n = 21) of the isolates were positive for the presence of intI1 gene. Analysis of gene cassettes revealed that six gene cassettes were found, which encoded resistance to trimethoprim (dhfr, dhfrI, dfrA17) and aminoglycosides (aadA1, aadA2, aadA5). Among them, the gene cassette array dfrA17–aadA5 was found most prevalent (66.7%). The resistance profile of positive-integron isolates was relatively broad and they were resistant to more than eight antimicrobials (n ? 8). The correlation analysis revealed the incidence of integrons among the isolates were related to the multiple antibiotic resistance profile, indicating integrons play an important role in the dissemination and spread of the antimicrobial resistant strains.     

Detection and characterisation of Shiga toxin-producing Escherichia coli other than Escherichia coli O157:H7 in wild ruminants

Shiga toxin-producing Escherichia coli (STEC) are an important group of emerging pathogens, with ruminants recognised as their main natural reservoir. The aim of this work was to establish the prevalence of non-O157 STEC in free-ranging wild ruminants in the Extremadura region of Spain and to characterise them phenogenotypically. Faecal samples were collected from 243 wild ruminants, including Cervus elaphus, Capreolus capreolus, Dama dama and Ovis musimon and were examined for STEC using both phenotypic (Vero cells) and genotypic (PCR and PFGE) methods.Shiga toxin-producing Escherichia coli were isolated from 58 (23.9%) of the samples and a total of 65 isolates were characterised. A PCR method indicated that 11 (16.9%) strains carried the stx₁ gene, 44 (67.7%) carried the stx₂ gene and 10 (15.4%) carried both these genes. The ehxA gene was detected in 37 (57%) of the isolates but none contained either the eae or saa genes. The isolates were from a total of 12 ‘O’ serogroups, although 80% were restricted to the O2, O8, O128, O146, O166 and O174 serogroups. The most commonly isolated STEC bacteria, which were from the O146 serogroup, exhibited a high degree of polymorphism as indicated by PFGE. Shiga toxin-producing Escherichia coli isolates of serogroups O20, O25, O166, O171, O174 and O176 had not previously been found in wild ruminants. This is the first study to confirm that wild ruminants in Spain are a reservoir of STEC and are thus a potential source of human infection.     

Candida peritonitis in dogs: report of 5 cases

Candida peritonitis is reported in people and is associated with significant morbidity and mortality compared with sterile or bacterial peritonitis. Recognized predisposing risk factors include peritoneal dialysis, hollow viscous organ perforation, abdominal surgery, inflamed intestinal mucosa, antimicrobial administration, and immunosuppression. In this report, we describe 5 cases of dogs with peritonitis complicated by Candida spp; 3 dogs with C albicans, one dog with C albicans and C glabrata, and one dog with C glabrata only. The 3 dogs with C albicans peritonitis presented with duodenal perforation due to NSAID therapy, intestinal resection and anastomosis following postspay‐surgery dehiscence, and intestinal foreign body removal. The 2 dogs with C glabrata peritonitis had undergone cholecystectomy due to gall bladder rupture and dehiscence of intestinal biopsy removal sites following exploratory laparatomy. In all cases, initial diagnosis of fungal peritonitis was made via cytologic examination of peritoneal effusions, which revealed marked pyogranulomatous inflammation with numerous 3–8 μm oval, deeply basophilic yeast organisms with thin clear capsules noted within phagocytes and extracellularly. In addition, germ tube formation, hyphae, and pseudohyphae were rarely seen in some of the cases with pure C albicans. Identity of the organisms was determined by culture in all cases and confirmed by PCR in 3 cases. Candida spp. are commensals normally inhabiting the alimentary, the upper respiratory, and the lower urogenital tracts of mammals. They are opportunistic pathogens that can invade and colonize tissue when a patient is immune‐compromised or there is disruption of the mucosal barrier. Candida peritonitis should be considered in patients with peritoneal contamination with gastrointestinal or biliary contents.     

Application of ERIC-PCR for the comparison of isolates of Haemophilus parasuis

Objective To validate a polymerase chain reaction (PCR) based method, Enterobacterial Repetitive Intergenic Consensus‐PCR (ERIC‐PCR), for the fingerprinting of Haemophilus parasuis strains and to use that method to differentiate isolates from apparently related outbreaks of Glässers disease on three pig farms. Design ERIC‐PCR was evaluated by comparing 15 different strains that represented all 15 recognised serovars in the Kielstein‐Rapp‐Gabrielson (KRG) scheme for serotyping H parasuis. Next, ERIC‐PCR was used to examine 14 Australian field isolates of H parasuis; 12 collected from three farms suffering apparently related outbreaks of Glässers disease and two from two other farms with no known connection. Results The 15 serovar reference strains all gave unique, reproducible ERIC‐PCR fingerprints. The 12 isolates from the three apparently related outbreaks all gave a single fingerprint, which was distinct from any seen in the 15 serovar reference strains and the two other Australian field isolates in the studied farms. The confirmation that all 12 isolates were the same strain allowed the development of a prevention and control program that has prevented the emergence of any further outbreaks of Glässer disease on the three farms. Conclusion ERIC‐PCR is a suitable technique for the differentiation of unrelated strains of H parasuis. The finding that the 12 field isolates of H parasuis all shared the same fingerprint is strong evidence that there was a common source of infection on all three farms. This study has shown, for the first time, that ERIC‐PCR is a suitable technique for the sub‐typing of H parasuis and useful for studying the epidemiology of outbreaks of Glässers disease.     

Multiplex-PCR-based differentiation and characterization of Candida-isolates derived from tortoises (Testudinidae)

Candida isolates (n=23) derived from Testudinidae were investigated by multiplex-polymerase chain reaction (PCR). The isolates comprised 13 Candida (C.) tropicalis, nine C. albicans and one C. parapsilosis. In addition, five reference strains C. albicans (ATCC 10231), C. tropicalis (DSM 4238), C. parapsilosis (DSM 4237), C. glabrata (ATCC 70614) and C. krusei (ATCC 70075) were investigated. PCR revealed easily distinguishable species-specific amplicons of the chs1-gene and resulted in a clear identification in all cases. No discrepancies between conventional identification techniques and the PCR-based system were seen. The described PCR offers a rapid alternative to conventional techniques for the identification of C. albicans, C. glabrata, C. tropicalis, C. parapsilosis and C. krusei.     

鸡源肺炎克雷伯菌的分离鉴定和耐药性分析

为探究鸡源肺炎克雷伯菌的流行性和耐药情况,本试验采集蛋鸡新鲜粪便60份(雏鸡36只/产蛋鸡24只),通过分离培养、VITEK2 Compact生化鉴定、特异性基因PCR扩增和微量肉汤稀释法对分离菌株进行了菌种鉴定、耐药表型、耐药基因以及毒力基因检测。结果显示,从粪便样本中共分离出48株肺炎克雷伯菌;分离菌株对氨苄西林、大观霉素、四环素、氟苯尼考、磺胺异噁唑和复方新诺明表现出高度耐药,耐药率范围为50.00%～100.00%,对奥格门丁、庆大霉素、头孢类和喹诺酮类药物耐药程度较低,耐药率范围为14.58%～27.08%,对黏菌素和美罗培南敏感;75.02%的菌株表现为多重耐药,最高表现为8重耐药,占10.42%。耐药基因和毒力基因检测结果显示,48株肺炎克雷伯菌共检出aadA1、tetA、oqxA、oqxB、bla_TEM和qnrB等6种耐药基因,以及entB、wabG、uge和kfuBC等4种毒力基因,本试验结果可为临床用药、动物源细菌耐药性监测和健康养殖提供数据支持。     

Virulence genes and plasmid profiles in Rhodococcus equi isolates from domestic pigs and wild boars (Sus scrofa) in Brazil

The virulence genes and plasmid profiles of 23 Rhodococcus equi isolates from 258 lymph nodes from domestic pigs (129 nodes with lesions and 129 without lesions) and 120 lymph nodes from slaughtered wild boars (60 nodes with lesions and 60 without) were characterized. R. equi was obtained from 19 lymph nodes of domestic pigs, 17 with, and two without lesions, and from four lymph nodes with lesions, from wild boars. The 23 isolates were tested for the presence of vapA and vapB genes, responsible for the 15–17 and 20 kDa virulence-associated proteins, respectively, by PCR in order to characterize as virulent (VapA), intermediately virulent (VapB) and avirulent. Plasmid DNAs were isolated and analyzed by digestion with restriction endonucleases to estimate size and compare their polymorphisms. Of the 19 domestic pigs strains, seven (36.8%) were avirulent and 12 (63.2%) were intermediately virulent, with the intermediately virulent isolates being plasmid types 8 (8 isolates), 10 (2 isolates), 1 (1 isolate) and 29 (1 isolate). The plasmid type of four strains isolated from wild boars was also intermediately virulent type 8. None of the domestic pigs and wild boar isolates showed the vapA gene. These findings demonstrate a high occurrence of plasmid type 8 in isolates from pigs and wild boars, and the similarity of plasmid types in the domestic pigs, wild boars and human isolates in Brazil.     

Detection and identification of food‐borne pathogens of the genera Campylobacter,Arcobacter and Helicobacter by multiplex PCR in poultry and poultry products

The objective of this study was to develop a multiplex polymerase chain reaction (PCR) to detect and differentiate food‐borne pathogens of the three genera Campylobacter, Arcobacter and Helicobacter in a single step procedure. One common reverse primer and three genus‐specific forward primers were designed by hybridizing to the 16S rRNA of selected reference strains. Besides the species with significance as food‐borne pathogens isolated from poultry meat –Campylobacter jejuni, Campylobacter coli, Arcobacter butzleri and Helicobacter pullorum– several other members of these genera were tested to determine the specificity of the designed multiplex PCR. In total, 20 ATCC and NCTC reference strains of Campyobacter, Arcobacter and Helicobacter were used to evaluate the PCR. Specific amplificates were obtained from all thermophilic species of Campylobacter as well as from species of Arcobacter and Helicobacter. No amplification product was obtained from the non‐thermophilic Campylobacter, C. hyointestinalis and C. fetus. Furthermore, a total of 43 field strains of the three genera isolated from poultry, pigs, cattle and humans were investigated using this PCR. To confirm the classification of 10 H. pullorum strains the 16S rRNAs were sequenced. The developed PCR is a helpful diagnostic tool to detect and differentiate Campylobacter, Arcobacter and Helicobacter isolated from poultry and poultry products.