Abstract
Ornithobacterium rhinotracheale is the etiological agent of chickens and turkeys’ respiratory diseases, reduction of eggs, growth retardation, and death. Present research aimed to conduct the isolation, recognition, and molecular investigations of the bacterium in commercial broiler chicken flocks in East Azerbaijan province, Northwest of Iran, by the partial sequencing of 16S rRNA gene, ERIC‑PCR, and RAPD‑PCR with the OPG11 and M13 primers. We obtained 330 specimens from tracheal swabs of 33 slaughtered broiler flocks, of which we found 14 isolates (4.24%) of five flocks (15.15%) to be O. rhinotracheale. Typing by RAPD assay with the OPG11 primer, and ERIC‑PCR, classified the isolates in two types of 1 and 2 molecular patterns, most of which belonged to type 1. However, the M13 primer‑based RAPD technique was inappropriate for distinguishing and categorizing the isolates and generating all of them in the same pattern. In a phylogenetic analysis of O. rhinotracheale based on 16S rRNA sequences, the strains generated three clusters (I‑III), in which all of the studied isolates fell in one cluster (cluster I). Based on the results obtained from the RAPD and ERIC‑PCR assays, the genetic patterns of broiler‑chicken‑isolated O. rhinotracheale strains in Northwestern Iran had no significant differences.
References
Allymehr M. 2006. Seroprevalence of Ornithobacterium rhinotracheale infection in broiler and broiler breeder chickens in West Azerbaijan province, Iran. J Vet Med A Physiol Pathol Clin Med, 53(1), 40-42.
Amonsin A., Wellehan J.F., Li L.L., Vandamme P., Lindemann C., Edman M., Robinson R. & Kapur V. 1997. Molecular epidemiology of Ornithobacterium rhinotracheale. J Clin Microbiol, 35, 2894-2898.
Asadpour Y., Banani M. & Pourbakhsh S.A. 2011. Isolation, identification and antibiotic sensitivity determination of Ornithobacterium rhinotracheale in slaughtering broiler chicken flocks of Guilan province. Iranian J Vet Res, 12, 345-349.
Banani M., Khaki P., Goodarzi H., Vandyousefi J. & Pourbakhsh S.A. 2000. Isolation and identification of Ornithobacterium rhinotracheale from a broiler and a pullet flock. Pajouhesh and Sazandegi, 46, 106-109. [In Persian]
Canal C.W., Leão J.A., Ferreira D.J., Macagnan M., Pippi Salle C.T. & Back A. 2003. Prevalence of antibodies against Ornithobacterium rhinotracheale in broilers and breeders in southern Brazil. Avian Dis, 47, 731-737.
Canal C.W., Leao J.A., Rocha S.L., Macagnan M., Lima-Rosa C.A., Oliveira S.D. & Back A. 2005. Isolation and characterization of Ornithobacterium rhinotracheale from chickens in Brazil. Res Vet Sci, 78, 225-230.
Chansiripornchai N., Wanasawaeng W. & Sasipreeyajan J. 2007. Seroprevalence and identification of Ornithobacterium rhinotracheale from broiler and broiler breeder flocks in Thailand. Avian Dis, 51, 777-780.
Chaslus-Dancla E., Lesage-Descauses M.C., Leroy-Setrin S., Martel J.L., Coudert P. & Lafont J.P. 1996. Validation of random amplified polymorphic DNA assays by ribotyping as tools for epidemiological surveys of Pasteurella from animals. Vet Microbiol, 52, 91-102.
Chou C.H., Lin S.Y., Chen C.L. & Tsai H.J. 2009. Use of random amplified polymorphic DNA analysis and single-enzyme amplified fragment length polymorphism in molecular typing of Ornithobacterium rhinotracheale strains. Avian Dis, 53, 108-114.
Erganis O., Hadimli H.H., Kav K., Çorlu M. & Ozturk D. 2002. A comparative study on detection of Ornithobacterium rhinotracheale antibodies in meat type turkeys by dot immunobinding assay, rapid agglutination test and serum agglutination test. Avian Pathol, 31, 201-204.
Erganis O., Hadimli H.H., Kav K. & Sayin Z. 2013. Phenotypic and molecular characterization of strains of Ornithobacterium rhinotracheale isolated from poultry in Turkey. African J Microbiol Res, 7, 82-88.
Hafez H. 2002. Diagnosis of Ornithobacterium rhinotracheale. Int J Poultry Sci, 1, 114-118.
Hall T.A. 1999. BioEdit: a user-friendly biological sequence alignment editor and analysis program for windows 95/98/NT. Nucleic Acids Symposium Series, 41, 95-98.
Hassanzadeh M., Karimi V., Fallah N. & Ashrafi I. 2010. Molecular characterization of Ornithobacterium rhinotracheale isolated from broiler chicken flocks of Iran. Turkish J Vet Animal Sci, 34, 373-378.
Joubert P., Higgins R., Laperle A., Mikaelian I., Venne D. & Silim A. 1999. Isolation of Ornithobacterium rhinotracheale from turkeys in Quebec, Canada. Avian Dis, 43, 622-626.
Koga Y. & Zavaleta A.I. 2005. Intraspecies genetic variability of Ornithobacterium rhinotracheale in commercial birds in Peru. Avian Dis, 49, 108-111.
Leroy-Setrin S., Lesage M.C., Chaslus-Dancla E. & Lafont J.P. 1995. Clonal diffusion of EPEC-like Escherichia coli from rabbits as detected by ribotyping and random amplified polymorphic DNA assays. Epidemiol Infect, 114, 113-121.
Leroy-Sétrin S., Flaujac G., Thénaisy K. & Chaslus-Dancla E. 1998. Genetic diversity of Ornithobacterium rhinotracheale strains isolated from poultry in France. Letters Appl Microbiol, 26, 189-193.
Markey B.K., Leonard F.C., Archambault M., Cullinane A. & Maguire D. 2013. Clinical Veterinary Microbiology. 2nd ed. The United Kingdom, CV Mosby Company, London, 404-405.
Mirzaie S. & Hassanzadeh M. 2013. Molecular and phylogenetic characterization of Ornithobacterium rhinotracheale isolates from turkey, quail, partridge and domestic pigeon in Iran. J Anim Poultry Sci, 2, 79-84.
Numee S., Hauck R. & Hafez H.M. 2012. Detection and typing of Ornithobacterium rhinotracheale from German poultry flocks. Avian Dis, 56, 654-658.
Olive D.M. & Bean P. 1999. Principles and applications of methods for DNA-based typing of microbial organisms. J Clin Microbiol, 37, 1661-1669.
Ozbey G., Ongor H., Balik D.T., Celik V., Kilic A. & Muz A. 2004. Investigations on Ornithobacterium rhinotracheale in broiler flocks in Elazig province located in the East of Turkey. Veterinární medicína, 49, 305-311.
Ozbey G., Ertas H.B. & Muz A. 2005. Random amplified polymorphic DNA (RAPD) analysis of Ornithobacterium rhinotracheale strains isolated from chickens in Turkey. Vet Med, 50, 526-530.
Rohlf F.J. 1997. NTSYS-pc: Numerical Taxonomy and Multivariate Analysis System version 2.02i. Applied Biostatistics Inc. Exeter Software, Setauket, New York.
Ryll M., Hinz K.H., Salisch H. & Kruse W. 1996. Pathogenicity of Ornithobacterium rhinotracheale for turkey poults under experimental conditions. Vet Rec, 139, 19.
Sakai E., Tokuyama Y., Nonaka F., Ohishi S., Ishikawa Y., Tanaka M. & Taneno A. 2000. Ornithobacterium rhinotracheale infection in Japan: Preliminary investigations. Vet Rec, 146, 502-503.
Sander A., Ruess M., Bereswill S., Schuppler M. & Steinbrueckner B. 1998. Comparison of different DNA fingerprinting techniques for molecular typing of Bartonella henselae isolates. J Clin Microbiol, 36, 2973-2981.
Schuijffel D.F., Van Empel P.C., Pennings A.M., Van Putten J.P. & Nuijten P.J. 2005. Successful selection of cross-protective vaccine candidates for Ornithobacterium rhinotracheale infection. Infect Immun, 73, 6812-6821.
Szabó R., Wehmann E., Makrai L., Nemes C., Gyuris E., Thuma A. & Magyar T. 2017. Characterization of Ornithobacterium rhinotracheale field isolates from Hungary. Avian Pathol, 46, 506–514.
Tamura K., Stecher G., Peterson D., Filipski A. & Kumar S. 2013. MEGA6: molecular evolutionary genetics analysis version 6.0. Mol Biol Evol, 30, 2725-2729.
Thachil A.J., Velayudhan B.T., Lopes-Berkas V.C., Halvorson D.A. & Nagaraja K.V. 2007. Application of polymerase chain reaction fingerprinting to differentiate Ornithobacterium rhinotracheale isolates. J Vet Diagnost Invest, 19, 417-420.
Thieme S., Mühldorfer K., Lüschow D., Hafez M.H. & Woo P.C.Y. 2016. Molecular characterization of the recently emerged poultry pathogen Ornithobacterium rhinotracheale by multilocus sequence typing. PLoS ONE, 11, e0148158.
Tsai H.J. & Huang C.W. 2006. Phenotypic and molecular characterization of isolates of Ornithobacterium rhinotracheale from chickens and pigeons in Taiwan. Avian Dis, 50, 502-507.
Turan N. & Ak S. 2002. Investigation of the presence of Ornithobacterium rhinotracheale in chickens in Turkey and determination of the seroprevalence of the infection using the enzyme-linked immunosorbent assay. Avian Dis, 46, 442-446.
Van Empel P., Van den Bosch H., Loeffen P. & Storm P. 1997. Identification and serotyping of Ornithobacterium rhinotracheale. J Clin Microbiol, 35, 418-421.
Van Empel P. & Hafez M.H. 1999. Ornithobacterium rhinotracheale: a review. Avian Pathol, 28, 217-227.