Mehdi VASFI MARANDI, Mansour BANANI, Naghmeh KABIRI, Fereshteh SABOURI, Mohsen BASHASHATI

Genetic analysis of Iranian Mycoplasma gallisepticum isolates using gene-targeted sequencing of complete pvpA gene

This study aims to evaluate Mycoplasma gallisepticum (MG) genetic diversity through the complete sequencing of the pvpAgene among six MG isolates present in industrial poultry from 2015 to 2016. The genetic comparison of these six isolates showed similaritiesof over 94% and 92% in terms of nucleotide and amino acid sequences, respectively. The size of the PvpA protein in the studiedisolates included three different sizes with 361, 364, and 366 amino acids. The carboxy-terminus of the studied PvpA proteins contained11 tetrapeptides of P-R-P-M/Q/N motifs with truncation in the proline-rich region. Based on the BLAST analysis, the pvpA genes of sixMGs were closely related to isolates from house finch in the USA and the S6 strain. According to the phylogenetic analysis, two isolateswere located along with the S6 strain that showed an outgroup association with MG isolates from Germany, China, and the USA. Otherisolates of this study were placed in a separate cluster with MGs isolated from Iran during 2006–2013. The results of this study are usefulto recognize the circulating genotypes in Iran and their differences from other MGs from different parts of the world. In addition,continuous monitoring of poultry flocks could be significant for obtaining valuable information related to different MG strains anddesigning effective control strategies.

PDF

___

1. Raviv Z, Ley DH. Mycoplasma gallisepticum infection. In: Swayne DE (editor). Diseases of Poultry. 13th ed. Ames, IA, USA: Iowa State University Press; 2013. pp. 877-893.
2. Kleven SH. Control of avian mycoplasma infections in commercial poultry. Avian Diseases 2008; 52 (3): 367-374.
3. Levisohn S, Kleven SH. Avian mycoplasmosis (Mycoplasma gallisepticum). Revue Scientifique et Technique 2000; 19 (2): 425-442.
4. Umar S, Munir MT, Ur-Rehman Z, Subhan S, Azam T et al. Mycoplasmosis in poultry: update on diagnosis and preventive measures. World’s Poultry Science Journal 2016; 73 (1): 17-28.
5. Khan MI, Lam KM, Yamamoto R. Mycoplasma gallisepticum strain variations detected by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Avian Diseases 1987; 31 (2): 315-320.
6. Kleven SH, Browning GF, Bulach DM, Ghiocas E, Morrow CJ et al. Examination of Mycoplasma gallisepticum strains using restriction endonuclease DNA analysis and DNA-DNA hybridisation. Avian Patholology 1988; 17 (3): 559-570.
7. Santha M, Lukacs K, Burg K, Bernath S, Rasko I et al. Intraspecies genotypic heterogeneity among Mycoplasma gallisepticum strains. Applied and Environmental Microbiology 1988; 54 (2): 607-609.
8. Adler HE, da Silva JM. Immunization against Mycoplasma gallisepticum. Avian Diseases 1970; 14 (4): 763-769.
9. Khan MI, Kirkpatrick BC, Yamamoto R. A Mycoplasma gallisepticum strain-specific DNA probe. Avian Diseases 1987; 31 (4): 907-909.
10. Nagai S, Kazama S, Yagihashi T. Ribotyping of Mycoplasma gallisepticum strains with a 16S ribosomal RNA gene probe. Avian Pathology 1995; 24 (4): 633-642.
11. Tigges E, Minion FC. Physical map of Mycoplasma gallisepticum. Journal of Bacteriology 1994; 176 (13): 4157-4159.
12. Barbour EK, Shaib HA, Jaber LS, Talhouk SN. Standardization and evaluation of random application of polymorphic DNA polymerase chain reaction in subspecies typing of Mycoplasma gallisepticum. International Journal of Applied Research in Veterinary Medicine 2005; 3 (2): 138-149.
13. Charlton BR, Bickford AA, Chin RP, Walker RL. Randomly amplified polymorphic DNA (RAPD) analysis of Mycoplasma gallisepticum isolates from turkeys from the central valley of California. Journal of Veterinary Diagnostic Investigation 1999; 11 (5): 408-415.
14. Charlton BR, Bickford AA, Walker RL, Yamamoto R. Complementary randomly amplified polymorphic DNA (RAPD) analysis patterns and primer sets to differentiate Mycoplasma gallisepticum strains. Journal of Veterinary Diagnostic Investigation 1999; 11 (2): 158-161.
15. Cherry JJ, Ley DH, Altizer S. Genotypic analyses of Mycoplasma gallisepticum isolates from songbirds by random amplification of polymorphic DNA and amplified-fragment length polymorphism. Journal of Wildlife Diseases 2006; 42 (2): 421- 428.
16. Fan HH, Kleven SH, Jackwood MW. Application of polymerase chain reaction with arbitrary primers to strain identification of Mycoplasma gallisepticum. Avian Diseases 1995; 39 (4): 729- 735.
17. Ferguson NM, Hepp D, Sun S, Ikuta N, Levisohn S, et al. Use of molecular diversity of Mycoplasma gallisepticum by gene-targeted sequencing (GTS) and random amplified polymorphic DNA (RAPD) analysis for epidemiological studies. Microbiology 2005; 151 (Pt. 6): 1883-1893.
18. Raviv Z, Callison S, Ferguson-Noel N, Laibinis V, Wooten R et al. The Mycoplasma gallisepticum 16S-23S rRNA intergenic spacer region sequence as a novel tool for epizootiological studies. Avian Diseases 2007; 51 (2): 555-560.
19. Razin S, Yogev D, Naot Y. Molecular biology and pathogenicity of mycoplasmas. Microbiology and Molecular Biology Reviews 1998; 62 (4): 1094-1156.
20. Boguslavsky S, Menaker D, Lysnyansky I, Liu T, Levisohn S et al. Molecular characterization of the Mycoplasma gallisepticum pvpA gene which encodes a putative variable cytadhesin protein. Infection and Immunity 2000; 68 (7): 3956-3964.
21. Noormohammadi AH. Role of phenotypic diversity in pathogenesis of avian mycoplasmosis. Avian Pathology 2007; 36 (6): 439-444.
22. Levisohn S, Rosengarten R, Yogev D. In vivo variation of Mycoplasma gallisepticum antigen expression in experimentally infected chickens. Veterinary Microbiology 1995; 45 (2-3): 219-231.
23. Alavinia SJ, Vasfi Marandi M, Bahonar A, Ghafouri SA, Alemohammad HS et al. Serological survey of Mycoplasma gallisepticum infection in broiler breeder farms in Mazandaran province by using RSPA and ELISA (through 2013). Journal of Veterinary Research 2016; 71 (3): 351-357 (in Persian with an abstract in English).
24. Rasoulinezhad S, Bozorgmehrifard MH, Hosseini H, Sheikhi N, Charkhkar S. Molecular detection and phylogenetic analysis of Mycoplasma gallisepticum from backyard and commercial turkey flocks in Iran. Veterinary Research Forum 2017; 8 (4): 293-298.
25. Valadan M, Pourbakhsh SA, Ghalenoei MR, Shokri G, Ahmadi AR et al. Serolagic and microbiologic identification of avian mycoplasmas from slaughtered poultry in Tehran. Veterinary Researches and Biological Products 2010; 88: 19-26 (in Persian with an abstract in English).
26. Frey ML, Hanson RP, Andrson DP. A medium for the isolation of avian mycoplasmas. American Journal of Veterinary Research. 1968; 29 (11): 2163-2171.
27. OIE. Avian Mycoplasmosis (Mycoplasma gallisepticum, M. synoviae). Manual of Diagnostic Tests and Vaccines for Terrestrial Animals. Paris, France: World Organisation for Animal Health; 2008.
28. Markham PF, Glew MD, Whithear KG, Walker ID. Molecular cloning of a member of the gene family that encodes pMGA, a hemagglutinin of Mycoplasma gallisepticum. Infection and Immunity 1993; 61 (3): 903-909.
29. Hall TA. BioEdit: A user-friendly biological sequence alignment editor and analysis program for Windows 95/98/NT. Nucleic Acids Symposium Series 1999; 41: 95-98.
30. Kumar S, Stecher G, Tamura K. MEGA7: Molecular Evolutionary Genetics Analysis Version 7.0 for bigger datasets. Molecular Biology and Evolution 2016; 33 (7): 1870-1874.
31. Tulman ER, Liao X, Szczepanek SM, Ley DH, Kutish GF et al. Extensive variation in surface lipoprotein gene content and genomic changes associated with virulence during evolution of a novel North American house finch epizootic strain of Mycoplasma gallisepticum. Microbiology 2012; 158 (Pt. 8): 2073- 2088.
32. Sohrab V, Baharsefat M. Mycoplasmosis in poultry in Iran. Annual Reports of Razi Institute 1955: 1955-1956.
33. Hernandez-Divers SM, Villegas P, Jimenez C, Hernandez-Divers SJ, Garcia M et al. Backyard chicken flocks pose a disease risk for neotropic birds in Costa Rica. Avian Diseases 2008; 52 (4): 558-566.
34. Kelly PJ, Chitauro D, Rohde C, Rukwava J, Majok A et al. Diseases and management of backyard chicken flocks in Chitungwiza, Zimbabwe. Avian Diseases 1994; 38 (3): 626-629.
35. MacArthur MW, Thornton JM. Influence of proline residues on protein conformation. Journal of Molecular Biology 1991; 218 (2): 397-412.
36. Williamson MP. The structure and function of proline-rich regions in proteins. Biochemical Journal 1994; 297 (Pt. 2): 249- 260.
37. Keeler CL Jr, Hnatow LL, Whetzel PL, Dohms JE. Cloning and characterization of a putative cytadhesin gene (mgc1) from Mycoplasma gallisepticum. Infection and Immunity 1996; 64 (5): 1541-1547.
38. Adler HE, Yamamoto R, Berg J. Strain differences of pleuropneumonialike organisms of avian origin. Avian Diseases 1957; 1 (1): 19-27.
39. Nunoya T, Kanai K, Yagihashi T, Hoshi S, Shibuya K et al. Natural case of salpingitis apparently caused by Mycoplasma gallisepticum in chickens. Avian Pathology 1997; 26 (2): 391-398.
40. Fischer JR, Stallknecht DE, Luttrell P, Dhondt AA, Converse KA. Mycoplasmal conjunctivitis in wild songbirds: the spread of a new contagious disease in a mobile host population. Emerging Infectious Diseases 1997; 3 (1): 69-72.
41. Staley M, Bonneaud C, McGraw KJ, Vleck CM, Hill GE. Detection of Mycoplasma gallisepticum in house finches (Haemorhous mexicanus) from Arizona. Avian Diseases 2018; 62 (1): 14-17.
42. Ley DH, Berkhoff JE, Levisohn S. Molecular epidemiologic investigations of Mycoplasma gallisepticum conjunctivitis in songbirds by random amplified polymorphic DNA analyses. Emerging Infectious Diseases 1997; 3 (3): 375-380.
43. Sprygin AV, Andreychuk DB, Elatkin NP, Zinyakov NG, Kolosov SN et al. Genetic diversity of Mycoplasma gallisepticum field isolates using partial sequencing of the pvpA gene fragment in Russia. Avian Diseases 2010; 54 (2): 899-904.