Molecular epidemiology of Mycoplasma hyorhinis porcine field isolates in the United States

Autoři: Maria J. Clavijo aff001;  Srinand Sreevatsan aff001;  Timothy J. Johnson aff002;  Albert Rovira aff001
Působiště autorů: Department of Veterinary Population Medicine, College of Veterinary Medicine, University of Minnesota, St Paul, Minnesota, United States of America aff001;  Department of Veterinary and Biomedical Sciences, University of Minnesota, St Paul, Minnesota, United States of America aff002
Vyšlo v časopise: PLoS ONE 14(10)
Kategorie: Research Article
doi: 10.1371/journal.pone.0223653


Mycoplasma hyorhinis is one of the causative agents of polyserositis and arthritis in post-weaning pigs. Here we describe the development of a multi-locus sequence typing (MLST) protocol for the characterization of M. hyorhinis field isolates. A total of 104 field isolates from different geographical locations, swine production systems, and clinical backgrounds, were analyzed. Twenty-seven genes, including housekeeping and those encoding surface proteins, were evaluated to index diversity. Genes encoding surface proteins were included to increase the discriminatory power of the MLST. Four target gene fragments were selected to be included in the final MLST-s (surface) protocol: pdhB, p95, mtlD and ung. Within each locus the nucleotide variation ranged from 1.4% to 20%. The 104 field isolates were classified into 39 distinct sequence types (STs). Multiple STs were found within the same production system and within the same pig. The majority of STs grouped strains from the same production system; however, cases existed where multiple systems shared a ST, indicating potential relationships between pig flows. The majority of the nucleotide changes observed in these genes generated synonymous changes, while non-synonymous changes were exclusively in the mtlD gene fragment, suggesting that this protein is undergoing selection. Molecular typing of M. hyorhinis will primarily aid swine practitioners with pig flow management and identifying sources of infection during outbreaks.

Klíčová slova:

Bronchi – Genetic loci – Genetic polymorphism – Nucleotide sequencing – Pleurae – Swine – Pericardium – Nasal cavity


1. Rovira A, Clavijo MJ, Oliveira S. Mycoplasma hyorhinis infection of pigs. Acta Sci Vet. 2010;38(Suppl 1): s9–15.

2. Clavijo MJ, Murray D, Oliveira S, Rovira A. Infection dynamics of Mycoplasma hyorhinis in three commercial pig populations. Vet Rec. 2017 Jul 15;18(3): 68. doi: 10.1136/vr.104064

3. Kobisch M, Friis NF. Swine mycoplasmoses. Rev Sci Tech. 1996 Dec;15(4): 1569–605. doi: 10.20506/rst.15.4.983 9190026

4. Neto JG, Gauger PC, Strait EL, Boyes N, Madson DM, Schwartz KJ. Mycoplasma-associated arthritis: critical points for diagnosis. J Swine Health Prod. 2012 Mar 1;20(2): 82–6.

5. Goiš M, Pospíšil Z, Černy M, Mrva V. Production of pneumonia after intranasal inoculation of gnotobiotic piglets with three strains of Mycoplasma hyorhinis. J Comp Pathol. 1971 Jul 1;81(3): 401–10. doi: 10.1016/0021-9975(71)90028-4 4935555

6. Friis NF. A serologic variant of Mycoplasma hyorhinis recovered from the conjunctiva of swine. Acta Veterinaria Scandinavica. 1976;17(3): 343–53. 983893

7. Friis NF. Mycoplasmas cultivated from the respiratory tract of Danish pigs. Acta Veterinaria Scandinavica. 1971;12(1): 69–79. 5575144

8. Rosengarten R, Wise KS. Phenotypic switching in mycoplasmas: phase variation of diverse surface lipoproteins. Science. 1990 Jan 19;247(4940): 315–8. doi: 10.1126/science.1688663 1688663

9. Schulman A, Estola T, Garry-Andersson AS. On the occurrence of Mycoplasma hyorhinis in the respiratory organs of pigs, with special reference to enzootic pneumonia. Zentralbl Veterinarmed B. 1970 May;17(5): 549–53. doi: 10.1111/j.1439-0450.1970.tb01565.x 5512059

10. Goiš M, Kuksa F. Intranasal infection of gnotobiotic piglets with Mycoplasma hyorhinis: differences in virulence of the strains and influence of age on the development of infection. Zoonoses Public Health. 1974 Jun 1;21(5): 352–61.

11. Kinne J, Johannsen U, Neumann R, Mehlhorn G, Pfützner H. The pathology and pathogenesis of experimental Mycoplasma hyorhinis infection of piglets with and without thermomotoric stress. 1. Pathologic-anatomic, histologic and immunomorphologic study results. Zentralbl Veterinarmed A. 1991 May;38(4): 306–20. 1907790

12. Clavijo MJ, Davies P, Morrison R, Bruner L, Olson S, Rosey E, et al. Temporal patterns of colonization and infection with Mycoplasma hyorhinis in two swine production systems in the US. Vet Microbiol. 2019 July 234 p 110–118. doi: 10.1016/j.vetmic.2019.05.021 31213266

13. Zadoks RN, Schukken YH. Use of molecular epidemiology in veterinary practice. Vet Clin North Am Food Anim Pract. 2006 Mar 1;22(1): 229–61. doi: 10.1016/j.cvfa.2005.11.005 16517304

14. Tankouo-Sandjong B, Sessitsch A, Liebana E, Kornschober C, Allerberger F, Hächler H, et al. MLST-v, multilocus sequence typing based on virulence genes, for molecular typing of Salmonella enterica subsp. enterica serovars. J Microbiol Methods. 2007 Apr 30;69(1): 23–36. doi: 10.1016/j.mimet.2006.11.013 17208323

15. Barlev NA, Borchsenius SN. Continuous distribution of Mycoplasma genome 443 sizes. Biomed. Sci. 1991;2: 641–645. 1841633

16. Yamaguti M, Oliveira RC, Marques LM, Buzinhani M, Buim MR, Neto RL, et al. Molecular characterization of Mycoplasma hyorhinis isolated from pigs using pulsed-field gel electrophoresis and 16S rRNA sequencing. Vet Record Open. 2015 Nov 30;2(2): e000093. doi: 10.1136/vetreco-2014-000093

17. Tenover FC, Arbeit RD, Goering RV, Mickelsen PA, Murray BE, Persing DH, et al. Interpreting chromosomal DNA restriction patterns produced by pulsed-field gel electrophoresis: criteria for bacterial strain typing. J Clin Microbiol. 1995 Sep;33(9): 2233–9. 7494007

18. King SJ, Leigh JA, Heath PJ, Luque I, Tarradas C, Dowson CG, et al. Development of a multilocus sequence typing scheme for the pig pathogen Streptococcus suis: identification of virulent clones and potential capsular serotype exchange. J Clin Microbiol. 2002 Oct 1;40(10): 3671–80. doi: 10.1128/JCM.40.10.3671-3680.2002 12354864

19. Olvera A, Cerda-Cuellar M, Aragon V. Study of the population structure of Haemophilus parasuis by multilocus sequence typing. Microbiology. 2006 Dec 1;152(Pt 12): 3683–90. doi: 10.1099/mic.0.29254-0 17159221

20. Mayor D, Jores J, Korczak BM, Kuhnert P. Multilocus sequence typing (MLST) of Mycoplasma hyopneumoniae: a diverse pathogen with limited clonality. Vet Microbiol. 2008 Feb 5;127(1–2): 63–72. doi: 10.1016/j.vetmic.2007.08.010 17884308

21. Tocqueville V, Ferré S, Nguyen NH, Kempf I, Marois-Créhan C. Multilocus sequence typing of Mycoplasma hyorhinis strains identified by a real-time TaqMan PCR assay. J Clin Microbiol. 2014 May 1;52(5): 1664–71. doi: 10.1128/JCM.03437-13 24622092

22. Trüeb B, Catelli E, Luehrs A, Nathues H, Kuhnert P. Genetic variability and limited clonality of Mycoplasma hyorhinis in pig herds. Vet Microbiol. 2016 Aug 15;191: 9–14. doi: 10.1016/j.vetmic.2016.05.015 27374901

23. Maiden MC, Bygraves JA, Feil E, Morelli G, Russell JE, Urwin R, et al. Multilocus sequence typing: a portable approach to the identification of clones within populations of pathogenic microorganisms. Proc Natl Acad Sci U S A. 1998 Mar 17;95(6): 3140–5. doi: 10.1073/pnas.95.6.3140 9501229

24. Cooper JE, Feil EJ. Multilocus sequence typing–what is resolved? Trends Microbiol. 2004;12(8): 373–377. doi: 10.1016/j.tim.2004.06.003 15276613

25. Clavijo MJ, Oliveira S, Zimmerman J, Rendahl A, Rovira A. Field evaluation of a quantitative polymerase chain reaction assay for Mycoplasma hyorhinis. J Vet Diagn Invest. 2014 Nov;26(6): 755–60. doi: 10.1177/1040638714555175 25319032

26. Lefèvre PC, Jones GE, Ojo MO. Pulmonary mycoplasmoses of small ruminants. Rev Sci Tech Off int Epiz. 1987;6(3): 713–757.

27. Liu W, Fang L, Li S, Li Q, Zhou Z, Feng Z, et al. Complete genome sequence of Mycoplasma hyorhinis strain HUB-1. J Bacteriol. 2010 Nov 1;192(21): 5844–5. doi: 10.1128/JB.00946-10 20802032

28. Calcutt MJ, Foecking MF, Rosales RS, Ellis RJ, Nicholas RA. Genome sequence of Mycoplasma hyorhinis strain GDL-1. J Bacteriol. 2012 Apr;194(7): 1848. doi: 10.1128/JB.00033-12 22408248

29. Kornspan JD, Lysnyansky I, Kahan T, Herrmann R, Rottem S, Nir-Paz R. Genome analysis of a Mycoplasma hyorhinis strain derived from a primary human melanoma cell line. J Bacteriol. 2011 Sep 1;193(17): 4543–4. doi: 10.1128/JB.05505-11 21705582

30. Goodison S, Urquidi V, Kumar D, Reyes L, Rosser CJ. Complete genome sequence of Mycoplasma hyorhinis strain SK76. Genome Announc. 2013 Feb 28;1(1). pii: e00101–12. doi: 10.1128/genomeA.00101-12 23405353

31. Darling AE, Mau B, Perna NT. progressive Mauve: multiple genome alignment with gene gain, loss and rearrangement. PLoS One. 2010 Jun 25;5(6): e11147. doi: 10.1371/journal.pone.0011147 20593022

32. Siqueira FM, Thompson CE, Virginio VG, Gonchoroski T, Reolon L, Almeida LG, et al. New insights on the biology of swine respiratory tract mycoplasmas from a comparative genome analysis. BMC genomics. 2013 Mar 14;14(1): 175.

33. Citti C, Wise KS. Mycoplasma hyorhinis vlp gene transcription: critical role in phase variation and expression of surface lipoproteins. Mol Microbiol. 1995 Nov;18(4): 649–660. doi: 10.1111/j.1365-2958.1995.mmi_18040649.x 8817488

34. Citti C, Blanchard A. Mycoplasmas and their host: emerging and re-emerging minimal pathogens. Trends Microbiol. 2013 Apr;21(4): 196–203. doi: 10.1016/j.tim.2013.01.003 23419218

35. McAuliffe L, Gosney F, Hlusek M, De Garnica ML, Spergser J, Kargl M, et al. Multilocus sequence typing of Mycoplasma agalactiae. J Medical Microbiol. 2011 Jun 1;60(Pt 6): 803–11. doi: 10.1099/jmm.0.028159–0

36. Manso-Silván L, Dupuy V, Lysnyansky I, Ozdemir U, Thiaucourt F. Phylogeny and molecular typing of Mycoplasma agalactiae and Mycoplasma bovis by multilocus sequencing. Vet Microbiol. 2012 Dec 28;161(1–2): 104–12. doi: 10.1016/j.vetmic.2012.07.015 22841405

37. Razin S. Peculiar properties of mycoplasmas: the smallest self-replicating prokaryotes. FEMS microbiology letters. 1992 Dec 15;100(1–3): 423–31. doi: 10.1111/j.1574-6968.1992.tb14072.x 1478475

38. Liu W, Xiao S, Li M, Guo S, Li S, Luo R, et al. Comparative genomic analyses of Mycoplasma hyopneumoniae pathogenic 168 strain and its high-passaged attenuated strain. BMC genomics. 2013 Dec;14(1):80.

39. Manso-Silván L, Dupuy V, Chu Y, Thiaucourt F. Multi-locus sequence analysis of Mycoplasma capricolum subsp. capripneumoniae for the molecular epidemiology of contagious caprine pleuropneumonia. Vet Res. 2011 Jul 14;42(1): 86. doi: 10.1186/1297-9716-42-86

40. van Loo IH, Heuvelman KJ, King AJ, Mooi FR. Multilocus sequence typing of Bordetella pertussis based on surface protein genes. J Clin Microbiol. 2002 Jun 1;40(6): 1994–2001. doi: 10.1128/JCM.40.6.1994-2001.2002 12037054

41. dos Santos LF, Clavijo MJ, Sreevatsan S, Rovira A, Moreira MA, Pieters M. Genotyping of Mycoplasma hyorhinis using multiple-locus variable number tandem repeat analysis. J Microbiol Methods. 2015 Apr 30;111: 87–92. doi: 10.1016/j.mimet.2015.02.003 25661497

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