Skip to content
1887

Access Microbiology open research platform logo Access Microbiology

Volume 7, Issue 2

A comparative analysis of the gene regions of and isolates from commercial hen farms in Mexico Open Access

Abstract

Avian mycoplasmosis, caused by and , poses significant economic challenges due to respiratory issues, reduced egg production and soft eggshells. The variable lipoprotein haemagglutinin (VlhA) protein, crucial for pathogenicity, comprises conserved (MSPB) and variable (MSPA) regions. The aim of this study was to identify the conserved region of gene sequences in field strain. We examined sequences from field strains collected in central Mexico (Jalisco and Mexico City). Specifically, we analysed 124 deformed eggs and 10 laying hens from 9 farms with Hy-line and Bovans breeds. Using PCR targeting the and 16S rRNA genes, we characterized 24 field strains, 4 of which were and 20 of which were . We analysed the regions, based on the AF035624.1 reference sequence, with American Type Culture Collection strains as positive controls. Additionally, we validated the PCR with 20 negative samples from isolation without the need for cultivation. We identified two amplification regions: MSPB and MSPA. Bioanalysis revealed relationships between our field samples and avian sequences in GenBank, alongside similarities with lipoproteins present in PG8 and . Given the significance of the VlhA protein in pathogenicity and immune evasion, the identified conserved sequences hold potential as therapeutic targets and for phylogenetic studies.

  • Received:
  • Accepted:
  • Published Online:
Keyword(s): avian , lipoprotein and Mycoplasma
Funding
This study was supported by the:
  • CONAHCyT. BECA
    • Principal Award Recipient: MayaLinda
  • DGAPA-UNAM (Award PAPIIT IN 219621)
    • Principal Award Recipient: J. Trigo-TaveraFrancisco
  • DGAPA-UNAM (Award PAPITT-IN221818)
    • Principal Award Recipient: J. Trigo-TaveraFrancisco
© 2025 The Authors
  • This is an open-access article distributed under the terms of the Creative Commons Attribution License. This article was made open access via a Publish and Read agreement between the Microbiology Society and the corresponding author’s institution.
Loading

Article metrics loading...

/content/journal/acmi/10.1099/acmi.0.000760.v4
2025-02-21
2026-03-16

Metrics

Loading full text...

Full text loading...

/deliver/fulltext/acmi/7/2/acmi000760.v4.html?itemId=/content/journal/acmi/10.1099/acmi.0.000760.v4&mimeType=html&fmt=ahah

References

  1. Ferguson‐Noel N, Armour NK, Noormohammadi AH, El‐Gazzar M, Mycoplasmosis BJM. Diseases of Poultry, Wiley, 14th edn Hoboken, NJ: John Wiley & Sons; 2020 pp 907–965
     [Google Scholar]
  2. Yadav JP, Tomar P, Singh Y, Khurana SK. Insights on Mycoplasma gallisepticum and Mycoplasma synoviae infection in poultry: a systematic review. Anim Biotechnol 2022; 33:1711–1720 [View Article]
     [Google Scholar]
  3. Benoina D, Tadina T, Dorrer D. Natural infection of geese with Mycoplasma gallisepticum and Mycoplasma synoviae and egg transmission of the mycoplasmas. Avian Pathol 1988; 17:925–928 [View Article] [PubMed]
     [Google Scholar]
  4. Jeon E-O, Kim J-N, Lee H-R, Koo B-S, Min K-C et al. Eggshell apex abnormalities associated with Mycoplasma synoviae infection in layers. J Vet Sci 2014; 15:579–582 [View Article] [PubMed]
     [Google Scholar]
  5. Kleven S, Brabury J. Avian mycoplasmosis (Mycoplasma gallisepticum, M. synoviae). In Manual of Diagnostic Tests and Vaccines for Terrestrial Animals, 6th edn. Paris: World Organization for Animal Health (OIE); 2008 pp 482–496
     [Google Scholar]
  6. Stipkovits L, Kempf I. Mycoplasmoses in poultry. Rev Sci Tech 1996; 15:1495–1525 [View Article] [PubMed]
     [Google Scholar]
  7. Kursa O, Pakuła A, Tomczyk G, Paśko S, Sawicka A. Eggshell apex abnormalities caused by two different Mycoplasma synoviae genotypes and evaluation of eggshell anomalies by full-field optical coherence tomography. BMC Vet Res 2019; 15:1 [View Article] [PubMed]
     [Google Scholar]
  8. Al-Baqir A, Hassanin O, Al-Rasheed M, Ahmed MS, Mohamed MHA et al. Mycoplasmosis in poultry: an evaluation of diagnostic schemes and molecular analysis of Egyptian Mycoplasma gallisepticum strains. Pathogens 2023; 12:1131 [View Article] [PubMed]
     [Google Scholar]
  9. Noormohammadi AH, Markham PF, Whithear KG, Walker ID, Gurevich VA et al. Mycoplasma synoviae has two distinct phase-variable major membrane antigens, one of which is a putative hemagglutinin. Infect Immun 1997; 65:2542–2547 [View Article] [PubMed]
     [Google Scholar]
  10. Hong Y, García M, Leiting V, Bencina D, Dufour-Zavala L et al. Specific detection and typing of Mycoplasma synoviae strains in poultry with PCR and DNA sequence analysis targeting the hemagglutinin encoding gene vlhA. Avian Dis 2004; 48:606–616 [View Article] [PubMed]
     [Google Scholar]
  11. Bencina D, Drobnic-Valic M, Horvat S, Narat M, Kleven SH et al. Molecular basis of the length variation in the N-terminal part of Mycoplasma synoviae hemagglutinin. FEMS Microbiol Lett 2001; 203:115–123 [View Article] [PubMed]
     [Google Scholar]
  12. Wetzel AN, Lefevre KM, Raviv Z. Revised Mycoplasma synoviae vlhA PCRs. Avian Dis 2010; 54:1292–1297 [View Article] [PubMed]
     [Google Scholar]
  13. El-Gazzar MM, Wetzel AN, Raviv Z. The genotyping potential of the Mycoplasma synoviae vlhA gene. Avian Dis 2012; 56:711–719 [View Article] [PubMed]
     [Google Scholar]
  14. Bencina D. Haemagglutinins of pathogenic avian mycoplasmas. Avian Pathol 2002; 31:535–547 [View Article]
     [Google Scholar]
  15. May M, Dunne DW, Brown DR. A sialoreceptor binding motif in the Mycoplasma synoviae adhesin VlhA. PLoS One 2014; 9:e110360 [View Article] [PubMed]
     [Google Scholar]
  16. Eissa S. Preparation and evaluation of Mycoplasma gallisepticum and Mycoplasma synoviae recombinant vaccine. ZVJ 20191–10 [View Article]
     [Google Scholar]
  17. Xu B, Chen X, Lu F, Sun Y, Sun H et al. Comparative genomics of Mycoplasma synoviae and new targets for molecular diagnostics. Front Vet Sci 2021; 8:640067 [View Article] [PubMed]
     [Google Scholar]
  18. Khiari AB, Mardassi BBA. Characterization of the antigenic and functional domains of a Mycoplasma synoviae variant vlhA gene. Vet Microbiol 2012; 156:322–329 [View Article] [PubMed]
     [Google Scholar]
  19. Vasconcelos ATR, Ferreira HB, Bizarro CV, Bonatto SL, Carvalho MO et al. Swine and poultry pathogens: the complete genome sequences of two strains of Mycoplasma hyopneumoniae and a strain of Mycoplasma synoviae. J Bacteriol 2005; 187:5568–5577 [View Article]
     [Google Scholar]
  20. Pflaum K, Tulman ER, Canter J, Dhondt KV, Reinoso-Perez MT et al. The influence of host tissue on M. gallisepticum vlhA gene expression. Vet Microbiol 2020; 251:108891 [View Article] [PubMed]
     [Google Scholar]
  21. Rosengarten R, Citti C, Glew M, Lischewski A, Droesse M et al. Host-pathogen interactions in Mycoplasma pathogenesis: virulence and survival strategies of minimalist prokaryotes. Int J Med Microbiol 2000; 290:15–25 [View Article] [PubMed]
     [Google Scholar]
  22. Lorenc Z, Paśko S, Kursa O, Pakuła A, Sałbut L. Spectral technique for detection of changes in eggshells caused by Mycoplasma synoviae. Poult Sci 2019; 98:3481–3487 [View Article] [PubMed]
     [Google Scholar]
  23. Whitford H, Rosenbusch R, Lauerman L. Mycoplasmosis in Animals: Laboratory Diagnosis Ames: Iowa State University; 1994
     [Google Scholar]
  24. Gardella RS, DelGiudice RA. Hemagglutination, hemadsorption, and hemolysis. In Methods in Mycoplasmology New York: Academic Press; 1983 pp 379–384
     [Google Scholar]
  25. Lockaby SB, Hoerr FJ, Lauerman LH, Smith BF, Samoylov AM et al. Factors associated with virulence of Mycoplasma synoviae. Avian Dis 1999; 43:251–261 [View Article] [PubMed]
     [Google Scholar]
  26. Sambrook J, Russell W. Molecular Cloning: A Laboratory Manual, Vol. 1, 3rd edn New York: Cold Spring Harbor Press; 2001
     [Google Scholar]
  27. García M, Ikuta N, Levisohn S, Kleven SH. Evaluation and comparison of various PCR methods for detection of Mycoplasma gallisepticum infection in chickens. Avian Dis 2005; 49:125–132 [View Article] [PubMed]
     [Google Scholar]
  28. Kumar S, Stecher G, Li M, Knyaz C, Tamura K. MEGA X: molecular evolutionary genetics analysis across computing platforms. Mol Biol Evol 2018; 35:1547–1549 [View Article] [PubMed]
     [Google Scholar]
  29. Ojha KK, Mishra S, Singh VK. Computational molecular phylogeny: concepts and applications. In Singh DB, Pathak RK. eds Bioinformatics New York: Academic Press; 2022 pp 67–89
     [Google Scholar]
  30. Giram P, Bhutada P, Prajapati C, Koratkar SS, Patil S et al. Percent positivity and phylogenetic analysis of Mycoplasma gallisepticum and Mycoplasma synoviae in commercial poultry from the different States of India. Vet World 2022; 15:1843–1851 [View Article] [PubMed]
     [Google Scholar]
  31. Cisneros-Tamayo M, Kempf I, Coton J, Michel V, Bougeard S et al. Investigation on eggshell apex abnormality (EAA) syndrome in France: isolation of Mycoplasma synoviae is frequently associated with Mycoplasma pullorum. BMC Vet Res 2020; 16:271 [View Article] [PubMed]
     [Google Scholar]
  32. Pflaum K, Tulman ER, Beaudet J, Canter J, Geary SJ. Variable lipoprotein hemagglutinin A gene (vlhA) expression in variant Mycoplasma gallisepticum strains in vivo. Infect Immun 2018; 86:86 [View Article] [PubMed]
     [Google Scholar]
  33. Hammond PP, Ramírez AS, Morrow CJ, Bradbury JM. Development and evaluation of an improved diagnostic PCR for Mycoplasma synoviae using primers located in the haemagglutinin encoding gene vlhA and its value for strain typing. Vet Microbiol 2009; 136:61–68 [View Article] [PubMed]
     [Google Scholar]
  34. May M, Brown DR. Diversity of expressed vlhA adhesin sequences and intermediate hemagglutination phenotypes in Mycoplasma synoviae. J Bacteriol 2011; 193:2116–2121 [View Article] [PubMed]
     [Google Scholar]
  35. Kang HM, Choi JG, Kim KI, Park HY, Park CK et al. Genetic and antigenic characteristics of H4 subtype avian influenza viruses in Korea and their pathogenicity in quails, domestic ducks and mice. J Gen Virol 2013; 94:30–39 [View Article] [PubMed]
     [Google Scholar]
  36. Lysnyansky I, Ron Y, Sachse K, Yogev D. Intrachromosomal recombination within the vsp locus of Mycoplasma bovis generates a chimeric variable surface lipoprotein antigen. Infect Immun 2001; 69:3703–3712 [View Article] [PubMed]
     [Google Scholar]
  37. Orlov M, Garanina I, Fisunov GY, Sorokin A. Comparative analysis of Mycoplasma gallisepticum vlhA promoters. Front Genet 2018; 9:569 [View Article] [PubMed]
     [Google Scholar]
  38. Urano-Tashiro Y, Takahashi Y, Oguchi R, Konishi K. Two arginine residues of Streptococcus gordonii sialic acid-binding adhesin Hsa are essential for interaction to host cell receptors. PLoS One 2016; 11:e0154098 [View Article] [PubMed]
     [Google Scholar]
  39. Serebryakova MV, Demina IA, Galyamina MA, Kondratov IG, Ladygina VG et al. The acylation state of surface lipoproteins of mollicute Acholeplasma laidlawii. J Biol Chem 2011; 286:22769–22776 [View Article] [PubMed]
     [Google Scholar]
  40. Dawood A, Algharib SA, Zhao G, Zhu T, Qi M et al. Mycoplasmas as host pantropic and specific pathogens: clinical implications, gene transfer, virulence factors, and future perspectives. Front Cell Infect Microbiol 2022; 12:855731 [View Article] [PubMed]
     [Google Scholar]
  41. Diard M, Hardt WD. Evolution of bacterial virulence. FEMS Microbiol Rev 2017; 41:679–697 [View Article] [PubMed]
     [Google Scholar]
  42. Rivera I, Molina R, Lee M, Mobashery S, Hermoso JA. Orthologous and paralogous AmpD peptidoglycan amidases from gram-negative bacteria. Microb Drug Resist 2016; 22:470–476 [View Article] [PubMed]
     [Google Scholar]
  43. Ghosh S, O’Connor TJ. Beyond paralogs: the multiple layers of redundancy in bacterial pathogenesis. Front Cell Infect Microbiol 2017; 7:467 [View Article]
     [Google Scholar]
  44. Lo Presti A, Carannante A, Fazio C, Neri A, Vacca P et al. Stefanelli P. FHbp variants among meningococci of serogroup B in Italy: evolution and selective pressure, 2014-2017. PLoS One 2023; 18:e0277976 [View Article] [PubMed]
     [Google Scholar]
  45. Yiwen C, Yueyue W, Lianmei Q, Cuiming Z, Xiaoxing Y. Infection strategies of mycoplasmas: unraveling the panoply of virulence factors. Virulence 2021; 12:788–817 [View Article] [PubMed]
     [Google Scholar]
  46. Zhang G, Han L, Li Z, Chen Y, Li Q et al. Screening of immunogenic proteins and evaluation of vaccine candidates against Mycoplasma synoviae. NPJ Vaccines 2023; 8:121 [View Article] [PubMed]
     [Google Scholar]
  47. Koonin EV. Orthologs, paralogs, and evolutionary genomics. Annu Rev Genet 2005; 39:309–338 [View Article] [PubMed]
     [Google Scholar]
  48. Altenhoff AM, Glover NM, Dessimoz C. Inferring orthology and paralogy. Methods Mol Biol 2019; 1910:149–175 [View Article] [PubMed]
     [Google Scholar]
  49. Lipman CB. Orthogenesis in Bacteria. Am Nat 1922; 56:105–115 [View Article]
     [Google Scholar]
  50. Deng H, Zhang GQ, Lin M, Wang Y, Liu ZJ. Mining from transcriptomes: 315 single-copy orthologous genes concatenated for the phylogenetic analyses of Orchidaceae. Ecol Evol 2015; 5:3800–3807 [View Article] [PubMed]
     [Google Scholar]
  51. Cohen JE, Wang R, Shen RF, Wu WW, Keller JE. Comparative pathogenomics of Clostridium tetani. PLoS One 2017; 12:e0182909 [View Article] [PubMed]
     [Google Scholar]
/content/journal/acmi/10.1099/acmi.0.000760.v4
Loading
A comparative in silico analysis of the vlhA gene regions of Mycoplasma gallisepticum and Mycoplasma synoviae isolates from commercial hen farms in Mexico
Access Microbiology 7, 000760.v4 (2025); https://doi.org/10.1099/acmi.0.000760.v4
/content/journal/acmi/10.1099/acmi.0.000760.v4
/content/journal/acmi/10.1099/acmi.0.000760.v4
Loading

Data & Media loading...

Supplements

Supplementary material 1

PDF

Most cited Most Cited RSS feed

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An error occurred
Approval was partially successful, following selected items could not be processed due to error