Molecular detection and phylogenetic analysis of Mycoplasma gallisepticum from backyard and commercial turkey flocks in Iran

Document Type: Original Article

Authors

1 Department of Poultry and Obstetrics, Science and Research Branch, Islamic Azad University, Tehran, Iran

2 Department of Clinical Science, Faculty of Veterinary Medicine, Karaj Branch, Islamic Azad University, Alborz, Iran

Abstract

Mycoplasma gallisepticum (MG) is economically important pathogen of poultry causes airsacculitis and frequently infraorbital sinusitis in turkeys. Infections may remain without clinical signs, but they can make birds susceptible to secondary infections.This study was carried out for molecular detection and phylogenetic analysis of MG infections in commercial and backyard turkey flocks in some parts of Iran. A total number of 600 swab samples were collected from 18 commercial and 31 backyard turkey flocks. The PCR technique was performed for detecting 16S rRNA gene in the samples. Positive sample were subjected for sequencing of mgc2 gene. The results showed that 48.38% of backyard and 16.66% of commercial farms were positive for MG. These findings suggested the presence of MG in the commercial and backyard turkeys’ farms of Iran. The molecular analysis indicated high sequence similarity between some Iranian turkeys isolates with Indian and Pakistanian MG isolates. Furthermore, substitutions of MG nucleic acids and correlated amino acids sequences may lead to some antigenic modifications.

Keywords

Main Subjects

  1. Ferguson-Noel N, Noormohammadi AH. Mycoplasma synoviae Infection. In: Swayne DE (Ed). Diseases of poultry. 13th ed. Hoboken, USA: Wiley-Blackwell 2013; 875-907.
  2. Feberwee A, De Wit JJ, Landman WJ. Induction of eggshell apex abnormalities by Mycoplasma synoviae: field and experimental studies. Avian Pathol 2009; 38(1):77-85.
  3. Levisohn S, Kleven SH. Avian mycoplasmosis (Mycoplasma gallisepticum). Rev Sci Tech 2000; 19(2): 425-442.
  4. Ley DH, Berkhoff JE, Levisohn S. Molecular epidemiologic investigations of Mycoplasma gallisepticum conjunctivitis in songbirds by random amplified polymorphic DNA analyses. Emerging Infect Dis 1997; 3(3):375-380.
  5. Wyrzykowski B, Albaric O, Moreau S, et al. Retrospective study of Mycoplasma gallisepticum meningoencephalitis in six turkey flocks in western France. J Comp Pathol 2013; 148(2): 173-177.
  6. Gross WB. Factors affecting the development of respiratory disease complex in chickens. Avian Dis 1990; 1:607-610.
  7. Kleven SH. Mycoplasmas in the etiology of multifactorial respiratory disease. Poult Sci 1998; 77(8): 1146-1149.
  8. Bradley LD, Snyder DB, Van Deusen RA. Identification of species-specific and interspecies-specific poly-peptides of Mycoplasma gallisepticum and Mycoplasma synoviae. Am J Vet Res 1988; 49(4): 511-515.
  9. Ferguson-Noel N, Laibinis VA, Farrar M. Influence of swab material on the detection of Mycoplasma gallisepticum and Mycoplasma synoviae by real-time PCR. Avian Dis 2012; 56(2): 310-314.
  10. Zhao S, Yamamoto R. Detection of Mycoplasma synoviae by polymerase chain reaction. Avian Pathol 1993; 22(3): 533-542.
  11. Khan MI. Multiplex PCR of avian pathogenic mycoplasmas. In: Saches K, Frey J (Eds). PCR detect of microbial pathogens. New York, USA: Humana Press 2003; 223-230.
  12. Santha MI, Burg KO, Rasko IS, et al. A species-specific DNA probe for the detection of Mycoplasma gallisepticum. Infect Immun 1987; 55(11): 2857-2859.
  13. Hyman HC, Levisohn S, Yogev D, et al. DNA probes for Mycoplasma gallisepticum and Mycoplasma synoviae: Application in experimentally infected chickens. Vet Mic 1989; 20(4): 323-337.
  14. Nascimento ER, Pereira VL, Nascimento MG, et al. Avian mycoplasmosis update. Rev Bras Cienc Avic 2005; 7(1): 1-9.
  15. Kleven SH, Fulton RM, Garcia M, et al. Molecular characterization of Mycoplasma gallisepticum isolates from turkeys. Avian Dis. 2004; 48(3): 562-269.
  16. Lysnyansky I, Garcia M, Levisohn S. Use of mgc2-polymerase chain reaction–restriction fragment length polymorphism for rapid differentiation between field isolates and vaccine strains of Mycoplasma gallisepticum in Israel. Avian Dis 2005; 49(2): 238-245.
  17. Garcia M, Ikuta N, Levisohn S, et al. Evaluation and comparison of various PCR methods for detection of Mycoplasma gallisepticum infection in chickens. Avian Dis 2005; 49(1): 125-132.
  18. Altschul SF, Madden TL, Schäffer AA, et al. Gapped BLAST and PSI-BLAST: A new generation of protein database search programs. Nucleic Acids Res 1997; 25(17): 3389-3402.
  19. Larkin MA, Blackshields G, Brown NP, et al. Clustal W and Clustal X version 2.0. Bioinformatics 2007; 23(21): 2947-2948.
  20. Burland TG. DNASTAR’s Lasergene sequence analysis software. Bioinformatics methods and protocols. Madison, USA: Humana Press 1999; 71-91.
  21. Tamura K, Stecher G, Peterson D, et al. MEGA 6: Molecular evolutionary genetics analysis version 6.0. Mol Biol Evol 2013; 30(12): 2725-2729.
  22. Swayne DE, Glisson RJ, McDougald LR, et al. Mycoplasma gallisepticum infection. In: Swayne DE (Ed). Diseases of poultry. 13th ed. Ames, USA: Blackwell Publishing Professional 2013; 877-893.
  23. Rezaie, M. Isolation and molecular identification of mycoplasma pathogen from turkeys of Eastern Azarbaijan provinces. DVSc thesis. Science and Research Branch, Islamic Azad University. Tehran, Iran: 2008.
  24. Michiels T, Welby S, Vanrobaeys M, et al. Prevalence of Mycoplasma gallisepticum and Mycoplasma synoviae in commercial poultry, racing pigeons and wild birds in Belgium. Avian Pathol 2016; 45(2): 244-252.
  25. Feberwee A, Landman WJ. The successful implementation of Mycoplasma gallisepticum monitoring and control programmes in Dutch commercial poultry: A declining sero-incidence during an eleven year period. In proceedings: 19th Congress of the international organization for mycoplasmology. Toulouse, France. 2012; 153.
  26. Chandiramani NK, Roekel H, Olesiuk OM. Viability studies with Mycoplasma gallisepticum under different environmental conditions. Poult Sci 1966; 45(5): 1029-1044.
  27. McBride MD, Hird DW, Carpenter TE, et al. Health survey of backyard poultry and other avian species located within one mile of commercial California meat-turkey flocks. Avian Dis 1991; 1: 403-407.
  28. Nagai S, Kazama S, Yagihashi T. Ribotyping of Mycoplasma gallisepticum strains with a 16S ribosomal RNA gene probe. Avian Pathol 1995; 24(4): 633-642.
  29. Ewing ML, Kleven SH, Brown MB. Comparison of enzyme-linked immunosorbent assay and hemagglutination-inhibition for detection of antibody to Mycoplasma gallisepticum in commercial broiler, fair and exhibition, and experimentally infected birds. Avian Dis 1996; 40 (1): 13-22.
  30. Seifi S, Shirzad MR. Risk factors and seroprevalence of Mycoplasma gallisepticum infection in broiler breeder farms in Mazandaran province, north of Iran. Revue Med Vet 2012; 163(5): 215-218.
  31. Haghighi-Khoshkhoo P, Akbariazad G, Roohi M, et al. Seroprevalence of Mycoplasma gallisepticum and Mycoplasma synoviae infection in the commercial layer flocks of the centernorth of Iran. Afr J Microbiol Res 2011; 5(18): 2834-2837.
  32. Bradbury JM, Levisohn S. Experimental infections in poultry. In: Tully JG (Ed). Molecular and diagnostic procedures in mycoplasmology. San Diego, USA: Academic Press 1996; 361-370.
  33. Roussan DA, Haddad R, Khawaldeh G. Molecular survey of avian respiratory pathogens in commercial broiler chicken flocks with respiratory diseases in Jordan. Poult Sci 2008; 87(3): 444-448.
  34. Hnatow LL, Keeler CL, Tessmer LL, et al. Characterization of MGC2, a Mycoplasma gallisepticum cytadhesin with homology to the Mycoplasma pneumoniae 30-kilodalton protein P30 and Mycoplasma genitalium P32. Infect Immun 1998; 66(7): 3436-3442.
  35. Winner F, Rosengarten R, Citti C. In vitro cell invasion of Mycoplasma gallisepticum. Infect Immun 2000; 68(7): 4238-4244.
  36. Delaney NF, Balenger S, Bonneaud C, et al. Ultrafast evolution and loss of CRISPRs following a host shift in a novel wildlife pathogen, Mycoplasma gallisepticum. PLoS Gen 2012; 8(2): e1002511.