Banner
Current IssueEditorial BoardOnline First ArticlesArchive
Current IssueEditorial BoardOnline First ArticlesArchive

Research Article

volume 52 issue 12 (december 2018) : 1715-1720,   Doi: 10.18805/ijar.B-3442

Sequencing and Phylogenetic analysis of vlhA gene from Mycoplasma synoviae field isolates associated with Eggshell apex abnormality

P
P. Sumitha
K
K. Sukumar
S
S. Srivignesh
1Department of Veterinary Microbiology, Veterinary College and Research Institute, Namakkal-637 002, Tamil Nadu, India
Cite article:- Sumitha P., Sukumar K., Srivignesh S. (2018). Sequencing and Phylogenetic analysis of vlhA gene from Mycoplasma synoviae field isolates associated with Eggshell apex abnormality. Indian Journal of Animal Research. 52(12): 1715-1720. doi: 10.18805/ijar.B-3442.

ABSTRACT

Eggshell apex abnormality (EAA) associated with Mycoplasma synoviae is occurring regularly in many commercial layer chicken farms of Tamil Nadu. Choanal cleft swabs were collected from EAA affected poultry farms, subjected for isolation and molecular detection of M. synoviae. Among 16 farms investigated M. synoviae could be isolated in six farms. All the farms were positive for M. synoviae in PCR. Among six isolates three isolates (VC&RI-NKL-SV3, VC&RI-NKL- PV6, VC&RI-NKL-KV2) from severe clinical outbreak were sequenced for vlhA gene and compared with the available sequences from the Gene Bank (NCBI) using multiple sequence alignments software - Clustal Omega and Bio edit programs. Analysis showed that the VC&RI,NKL-SV3 and VC&RI-NKL-KV2 were more similar (94.3%) to each other. Point mutation percentage of isolates VC&RI-NKL-PV6, VC&RI-NKL-SV3 and VC&RI-NKL-KV2 isolates in comparison with reference strain KC506806 were 12, 5.1 and 1.8% respectively. The phylogenetic analysis indicated that the VC&RI-NKL-SV3 and VC&RI-NKL-PV6 were closely evolved with VC&RI-NKL-KV2. VC&RI-NKL-KV2 was closely related to sequence of Mycoplasma synoviae from Brazil.

REFERENCES

  1. Bencina D, Drobnic-Valic M, Horvat S, Narat M, Kleven S H and Dovc P. (2001). Molecular basis of the length variation in the N-    terminal part of Mycoplasma synoviae hemagglutinin. FEMS Microbiol. Lett., 203: 115–123.
  2. Buim M, Buzinhani M, Yamaguti M, Oliveira R, Mettifogo E, Timenetsky J and Ferreira A. (2010). Intraspecific variation in 16S rRNA gene of Mycoplasma synoviae determined by DNA sequencing. Comp. Immunol. Microbiol. Infect. Dis., 33 (1): 15-23.
  3. Catania S, Bilato D, Gobbo F, Granato A, Terregino C, Iob L and Nicholas R A J. (2010). Treatment of eggshell abnormalities and reduced egg production caused by Mycoplasma synoviae infection. Avian Dis., 54 (2): 961- 964.
  4. Feberwee A, Wit J J D and Landman W J M. (2009). Induction of eggshell apex abnormalities by Mycoplasma synoviae: field and experimental studies. Avian Pathol., 38: 77-85.
  5. Harada K, Kijima-Tanaka M, Uchiyama M, Yamamoto T, Oishi K, Areo M and Takahashi T. (2009). Molecular Typing of Japanese Field Isolates and live commercial vaccine strain of Mycoplasma synoviae using improved pulsed-field gel electrophoresis and vlhA gene sequencing. Avian Dis., 53: 538–543. 
  6. Hong Y, García M, Leiting L, Bencina D, Dufour-zavala L, Zavala G and Kleven S H. (2004). Specific detection and typing of Mycoplasma synoviae strains in poultry with PCR and DNA sequence analysis targeting the hemagglutinin encoding gene vlhA. Avian Dis., 48: 606–616.
  7. Jain N C, Chandiramani N K and Singh I P. (1971). Studies on avian pleuropneumonia like organisms. Isolation and identification of Mycoplasma gallisepticum. J. Res. Ludhiana., 8: 118-124.
  8. Jeffery N, Gasser R B, Steer P A and Noormohammadi A H. (2007). Classification of Mycoplasma synoviae strains using single-strand conformation polymorphism and high-resolution melting-curve analysis of the vlhA gene single-copy region. Microbiol., 153 (8): 2679–2688.
  9. Kleven S H. (2003). Mycoplasma synoviae infection. In: Y. M. Saif, H. J. Barnes, J. R. Glisson, A. M. Fadly, L. R. McDougald and D. E. Swayne, Diseases of Poultry(eds), 11th edn, Iowa State Press, Ames. Pp: 756-766.
  10. Liu T, Garcia M, Levisohn S, Yogev D and Kleven S H. (2001). Molecular variability of the adhesin-encoding gene pvpA among Mycoplasma gallisepticum strains and its application in diagnosis. J. Clini. Microbiol., 39 (5):1882-1888.
  11. Pourbakhsh S A, Shokri G R, Banani M, Elhamnia M and Ashtari A. (2010). Detection of Mycoplasma synoviae infection in broiler breeder farms of Tehran province using PCR and culture methods. Arch. Razi. Inst., 65 (2): 75-81.
  12. Pourbakhsh S A, Maghami M, Ashtari A, Bayatzadesh M A and Ahangaran S. (2013). The vlhA gene sequencing of Iranian Mycoplasma synoviae isolates. Arch. Razi. Inst., 68 (2): 117-124.
  13. Ramadass, P, Ananthi R, Senthilkumar T M A, Venkatesh G and Ramaswamy V. (2006). Isolation and characterization of Mycoplasma gallisepticum and Mycoplasma synoviae from poultry. Indian J. Anim. Sci., 76 (10): 796-798.
  14. Razin S, Yogev D and Naot Y. (1998). Molecular biology and pathogenicity of mycoplasmas. Microbiol. Mol. Biol. rev., 62: 1094-156.
  15. Strugnell B W, Mcmullin P, Wood A M, Nicholas R A J, Ayling R and Irvine R M. (2011). Unusual eggshell defects in a free-range layer flock in Great Britain. Vet. Rec., 169: 237-238.
  16. Sumitha P, Moorthy M, Sukumar K, Mani K and Arthanari Eswaran M. (2015). Prevalence of Ornithobacterium rhinotracheale, Mycoplasma gallisepticum and Mycoplasma synoviae Antibodies in Layers. Indian Vet. J., 92 (8) : 78 - 79. 
Published In
Indian Journal of Animal Research
Article Metrics
Metrics Icon
0
Views
0
Citations
Reviewed By
Share Article
Download Article
In this Article
    Contact us
    Follow us

    Research Article

    volume 52 issue 12 (december 2018) : 1715-1720,   Doi: 10.18805/ijar.B-3442

    Sequencing and Phylogenetic analysis of vlhA gene from Mycoplasma synoviae field isolates associated with Eggshell apex abnormality

    P
    P. Sumitha
    K
    K. Sukumar
    S
    S. Srivignesh
    1Department of Veterinary Microbiology, Veterinary College and Research Institute, Namakkal-637 002, Tamil Nadu, India
    Cite article:- Sumitha P., Sukumar K., Srivignesh S. (2018). Sequencing and Phylogenetic analysis of vlhA gene from Mycoplasma synoviae field isolates associated with Eggshell apex abnormality. Indian Journal of Animal Research. 52(12): 1715-1720. doi: 10.18805/ijar.B-3442.

    ABSTRACT

    Eggshell apex abnormality (EAA) associated with Mycoplasma synoviae is occurring regularly in many commercial layer chicken farms of Tamil Nadu. Choanal cleft swabs were collected from EAA affected poultry farms, subjected for isolation and molecular detection of M. synoviae. Among 16 farms investigated M. synoviae could be isolated in six farms. All the farms were positive for M. synoviae in PCR. Among six isolates three isolates (VC&RI-NKL-SV3, VC&RI-NKL- PV6, VC&RI-NKL-KV2) from severe clinical outbreak were sequenced for vlhA gene and compared with the available sequences from the Gene Bank (NCBI) using multiple sequence alignments software - Clustal Omega and Bio edit programs. Analysis showed that the VC&RI,NKL-SV3 and VC&RI-NKL-KV2 were more similar (94.3%) to each other. Point mutation percentage of isolates VC&RI-NKL-PV6, VC&RI-NKL-SV3 and VC&RI-NKL-KV2 isolates in comparison with reference strain KC506806 were 12, 5.1 and 1.8% respectively. The phylogenetic analysis indicated that the VC&RI-NKL-SV3 and VC&RI-NKL-PV6 were closely evolved with VC&RI-NKL-KV2. VC&RI-NKL-KV2 was closely related to sequence of Mycoplasma synoviae from Brazil.

    REFERENCES

    1. Bencina D, Drobnic-Valic M, Horvat S, Narat M, Kleven S H and Dovc P. (2001). Molecular basis of the length variation in the N-    terminal part of Mycoplasma synoviae hemagglutinin. FEMS Microbiol. Lett., 203: 115–123.
    2. Buim M, Buzinhani M, Yamaguti M, Oliveira R, Mettifogo E, Timenetsky J and Ferreira A. (2010). Intraspecific variation in 16S rRNA gene of Mycoplasma synoviae determined by DNA sequencing. Comp. Immunol. Microbiol. Infect. Dis., 33 (1): 15-23.
    3. Catania S, Bilato D, Gobbo F, Granato A, Terregino C, Iob L and Nicholas R A J. (2010). Treatment of eggshell abnormalities and reduced egg production caused by Mycoplasma synoviae infection. Avian Dis., 54 (2): 961- 964.
    4. Feberwee A, Wit J J D and Landman W J M. (2009). Induction of eggshell apex abnormalities by Mycoplasma synoviae: field and experimental studies. Avian Pathol., 38: 77-85.
    5. Harada K, Kijima-Tanaka M, Uchiyama M, Yamamoto T, Oishi K, Areo M and Takahashi T. (2009). Molecular Typing of Japanese Field Isolates and live commercial vaccine strain of Mycoplasma synoviae using improved pulsed-field gel electrophoresis and vlhA gene sequencing. Avian Dis., 53: 538–543. 
    6. Hong Y, García M, Leiting L, Bencina D, Dufour-zavala L, Zavala G and Kleven S H. (2004). Specific detection and typing of Mycoplasma synoviae strains in poultry with PCR and DNA sequence analysis targeting the hemagglutinin encoding gene vlhA. Avian Dis., 48: 606–616.
    7. Jain N C, Chandiramani N K and Singh I P. (1971). Studies on avian pleuropneumonia like organisms. Isolation and identification of Mycoplasma gallisepticum. J. Res. Ludhiana., 8: 118-124.
    8. Jeffery N, Gasser R B, Steer P A and Noormohammadi A H. (2007). Classification of Mycoplasma synoviae strains using single-strand conformation polymorphism and high-resolution melting-curve analysis of the vlhA gene single-copy region. Microbiol., 153 (8): 2679–2688.
    9. Kleven S H. (2003). Mycoplasma synoviae infection. In: Y. M. Saif, H. J. Barnes, J. R. Glisson, A. M. Fadly, L. R. McDougald and D. E. Swayne, Diseases of Poultry(eds), 11th edn, Iowa State Press, Ames. Pp: 756-766.
    10. Liu T, Garcia M, Levisohn S, Yogev D and Kleven S H. (2001). Molecular variability of the adhesin-encoding gene pvpA among Mycoplasma gallisepticum strains and its application in diagnosis. J. Clini. Microbiol., 39 (5):1882-1888.
    11. Pourbakhsh S A, Shokri G R, Banani M, Elhamnia M and Ashtari A. (2010). Detection of Mycoplasma synoviae infection in broiler breeder farms of Tehran province using PCR and culture methods. Arch. Razi. Inst., 65 (2): 75-81.
    12. Pourbakhsh S A, Maghami M, Ashtari A, Bayatzadesh M A and Ahangaran S. (2013). The vlhA gene sequencing of Iranian Mycoplasma synoviae isolates. Arch. Razi. Inst., 68 (2): 117-124.
    13. Ramadass, P, Ananthi R, Senthilkumar T M A, Venkatesh G and Ramaswamy V. (2006). Isolation and characterization of Mycoplasma gallisepticum and Mycoplasma synoviae from poultry. Indian J. Anim. Sci., 76 (10): 796-798.
    14. Razin S, Yogev D and Naot Y. (1998). Molecular biology and pathogenicity of mycoplasmas. Microbiol. Mol. Biol. rev., 62: 1094-156.
    15. Strugnell B W, Mcmullin P, Wood A M, Nicholas R A J, Ayling R and Irvine R M. (2011). Unusual eggshell defects in a free-range layer flock in Great Britain. Vet. Rec., 169: 237-238.
    16. Sumitha P, Moorthy M, Sukumar K, Mani K and Arthanari Eswaran M. (2015). Prevalence of Ornithobacterium rhinotracheale, Mycoplasma gallisepticum and Mycoplasma synoviae Antibodies in Layers. Indian Vet. J., 92 (8) : 78 - 79. 
    In this Article
      Contact us
      Follow us
      Published In
      Indian Journal of Animal Research

      Editorial Board

      View all (0)