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Abstract：
BACKGROUND: Ornithobacterium rhinotracheal (ORT) infects numerous birds, particularly chickens and turkeys. ORT is an emerging bacterial pathogen of global concern in the poultry industry. As ORT is rapidly spreading throughout commercial poultry, it requires intensive studies of its epidemiology, diagnostic procedures, molecular typing, virulence genes and antimicrobial resistance.
OBJECTIVE: The present study was conducted in isolation and identification of ORT from slaughtered turkeys.
METHODS: Cleft palate swabs of 200 were collected from slaughtered turkeys and cultured on blood agar. ORT was characterized using biochemical tests and PCR targeting the ORT 16S rRNA gene. Virulence genes of isolates were determined targeting adenylate kinase (adk), copA and virulence-associated protein D (vapD) genes. Additionally, diversity of ORT isolates was performed by enterobacterial repetitive intergenic consensus (ERIC) and RAPD PCR. Disk diffusion was used to determine the antibiotic sensitivity of the isolates.
RESULTS: ORT was identified in 23 (11.5%) samples using both the biochemical tests and PCR. The result of detecting virulence genes showed that all the isolates (23: 100%) had the adk gene, whereas two (8.7%) isolates had the copA gene, and seven (30.43%) isolates had the vapD gene. Molecular typing of isolates revealed 21 different patterns by RAPD PCR assay using M13 primer and 20 distinct patterns by ERIC PCR test. Both ERIC and RAPD PCR were distinctive methods for investigating the genetic diversity of ORT isolates. The antibiotic resistance test showed that 18 (78.26%) isolates were resistant to gentamicin, amikacin, cefazolin, streptomycin and penicillin. All isolates (100%) were resistant to cloxacillin and fosfomycin.
CONCLUSIONS: This study showed the prevalence of ORT in turkey and high resistance of this bacterium to many common veterinary antibiotics. Moreover, both ERIC and RAPD PCR are distinctive methods for investigating the genetic diversity of ORT isolates. These data may help monitor antibiotic resistance and typing of ORT in epidemiological studies and serve as the foundation for designing region-specific vaccines for future use.
OBJECTIVE: The present study was conducted in isolation and identification of ORT from slaughtered turkeys.
METHODS: Cleft palate swabs of 200 were collected from slaughtered turkeys and cultured on blood agar. ORT was characterized using biochemical tests and PCR targeting the ORT 16S rRNA gene. Virulence genes of isolates were determined targeting adenylate kinase (adk), copA and virulence-associated protein D (vapD) genes. Additionally, diversity of ORT isolates was performed by enterobacterial repetitive intergenic consensus (ERIC) and RAPD PCR. Disk diffusion was used to determine the antibiotic sensitivity of the isolates.
RESULTS: ORT was identified in 23 (11.5%) samples using both the biochemical tests and PCR. The result of detecting virulence genes showed that all the isolates (23: 100%) had the adk gene, whereas two (8.7%) isolates had the copA gene, and seven (30.43%) isolates had the vapD gene. Molecular typing of isolates revealed 21 different patterns by RAPD PCR assay using M13 primer and 20 distinct patterns by ERIC PCR test. Both ERIC and RAPD PCR were distinctive methods for investigating the genetic diversity of ORT isolates. The antibiotic resistance test showed that 18 (78.26%) isolates were resistant to gentamicin, amikacin, cefazolin, streptomycin and penicillin. All isolates (100%) were resistant to cloxacillin and fosfomycin.
CONCLUSIONS: This study showed the prevalence of ORT in turkey and high resistance of this bacterium to many common veterinary antibiotics. Moreover, both ERIC and RAPD PCR are distinctive methods for investigating the genetic diversity of ORT isolates. These data may help monitor antibiotic resistance and typing of ORT in epidemiological studies and serve as the foundation for designing region-specific vaccines for future use.
摘要：
背景：鼻气管（ORT）感染许多鸟类，尤其是鸡和火鸡。ORT是家禽业中全球关注的新兴细菌病原体。由于ORT在商业家禽中迅速传播，它需要对其流行病学进行深入研究，诊断程序,分子分型,毒力基因和抗菌素耐药性。
目的：本研究从屠宰火鸡中分离鉴定ORT。
方法：从屠宰的火鸡中收集200只腭裂拭子，并在血琼脂上培养。使用生化测试和针对ORT16SrRNA基因的PCR对ORT进行表征。确定分离株的毒力基因靶向腺苷酸激酶（adk），copA和毒力相关蛋白D(vapD)基因。此外,ORT分离株的多样性是通过肠细菌重复基因间共识（ERIC）和RAPDPCR进行的。使用圆盘扩散来确定分离物的抗生素敏感性。
结果：使用生化测试和PCR在23个（11.5％）样品中鉴定出ORT。毒力基因检测结果表明,所有分离株(23:100%)均具有adk基因,而两个（8.7％）分离株具有copA基因，和7个（30.43％）分离株具有vapD基因。分离株的分子分型通过使用M13引物的RAPDPCR分析揭示了21种不同的模式，通过ERICPCR测试揭示了20种不同的模式。ERIC和RAPDPCR都是研究ORT分离株遗传多样性的独特方法。抗生素耐药性试验显示18株(78.26%)对庆大霉素耐药,阿米卡星，头孢唑啉,链霉素和青霉素。所有分离株（100％）对氯唑西林和磷霉素具有抗性。
结论：这项研究表明ORT在火鸡中的流行，并且该菌对许多常用兽用抗生素具有较高的耐药性。此外，ERIC和RAPDPCR都是研究ORT分离株遗传多样性的独特方法。这些数据可能有助于在流行病学研究中监测抗生素耐药性和ORT分型，并作为设计未来使用的区域特异性疫苗的基础。
目的：本研究从屠宰火鸡中分离鉴定ORT。
方法：从屠宰的火鸡中收集200只腭裂拭子，并在血琼脂上培养。使用生化测试和针对ORT16SrRNA基因的PCR对ORT进行表征。确定分离株的毒力基因靶向腺苷酸激酶（adk），copA和毒力相关蛋白D(vapD)基因。此外,ORT分离株的多样性是通过肠细菌重复基因间共识（ERIC）和RAPDPCR进行的。使用圆盘扩散来确定分离物的抗生素敏感性。
结果：使用生化测试和PCR在23个（11.5％）样品中鉴定出ORT。毒力基因检测结果表明,所有分离株(23:100%)均具有adk基因,而两个（8.7％）分离株具有copA基因，和7个（30.43％）分离株具有vapD基因。分离株的分子分型通过使用M13引物的RAPDPCR分析揭示了21种不同的模式，通过ERICPCR测试揭示了20种不同的模式。ERIC和RAPDPCR都是研究ORT分离株遗传多样性的独特方法。抗生素耐药性试验显示18株(78.26%)对庆大霉素耐药,阿米卡星，头孢唑啉,链霉素和青霉素。所有分离株（100％）对氯唑西林和磷霉素具有抗性。
结论：这项研究表明ORT在火鸡中的流行，并且该菌对许多常用兽用抗生素具有较高的耐药性。此外，ERIC和RAPDPCR都是研究ORT分离株遗传多样性的独特方法。这些数据可能有助于在流行病学研究中监测抗生素耐药性和ORT分型，并作为设计未来使用的区域特异性疫苗的基础。
