Swine hemoplasmosis detected in farms of Argentina by a new nested PCR assay

M. E. Pintos; D. M. Posik; E. Bonzo; C. J. Perfumo; M. S. Arauz

doi:10.30972/vet.3225742

Autores/as

M. E. Pintos Universidad Nacional del Nordeste
D. M. Posik Universidad Nacional de La Plata
E. Bonzo Universidad Nacional de La Plata
C. J. Perfumo Universidad Nacional de La Plata
M. S. Arauz Universidad Nacional de La Plata

DOI:

https://doi.org/10.30972/vet.3225742

Palabras clave:

Pig, hemoplasma, nested PCR, Argentina

Resumen

Swine hemoplasmosis or swine infectious anemia is a worldwide distribution disease caused by the hemotropic mycoplasmas Mycoplasma suis and Mycoplasma parvum. The aim of this study was to determine the presence of M. suis-M. parvum infection in subclinical pigs from herds of Buenos Aires province, Argentina, by means of new nested PCR. The PCR assay primers were designed based on the 16S ribosomal gene sequences of swine hemoplasmas available at GenBank. To standardize the assay, pig blood samples positive for hemoplasma by May Grünwald-Giemsa (MGG) stained blood smears were used. A total of 482 pig blood samples were analyzed. A 32% (154/482) of MGG stained blood smears were positive to M. suis o M. parvum. From these 154 samples, 47% (72/154) were positive by PCR. Sequences of PCR products amplified with these primers always showed identity with M. suisor M. parvum, validating their specificity and highlighting the unspecific amplification with hemoplasmas of other species. This is the first assay designed in Argentina to identify M. suis and M. parvum. However, considering the advances in the knowledge of the genome of hemoplasmas worldwide, further studies should be performed to standardize new assays for the diagnosis of swine hemoplasmosis in Argentina.

Descargas

Los datos de descargas todavía no están disponibles.

Citas

Anziani OS, Ford CA, Tarabla HD. 1986. Eperythrozoonosis porcina en la República Argentina. Rev Med Vet 67: 99-101.

Cornish-Bowden. Nomenclature for incompletely specified bases in nucleic acid sequences recommendations 1985. Nucleic Acids Res 13: 9, 3021-3030.

Dirienzo JA et al. Software estadístico. InfoStat versión 2018. Support@infostat.com.ar.

Donascimento NC et al. 2013. Genome sequence of Mycoplasma parvum (formerly Eperythrozoon parvum), a diminutive hemoplasma of the pig. Genome Announc. https://áoi.org/10.1128/genomeA.00986-13.

Donascimento NC et al. 2014. Microscopy and genomic analysis of Mycoplasma parvum strain Indiana. Vet Res 13: 45, 86.

Fu Y et al. 2017. Identification of a novel hemoplasma species from pigs in Zhejiang Province, China. J Vet Med Sci 79: 5, 864-870.

Groebel K, Hoelzle K, Wittenbrink MM, Ziegler U, Hoelzle LE. 2009. Mycoplasma suis invades porcine erythrocytes. InfectImm un 77: 2, 576-584.

Hall TA. 1999. BioEdit: a user-friendly biological sequence alignment editor and analysis program for Windows 95/98/NT. Nucl Acids Symp Ser 41: 95-98.

Hoelzle LE, Adelt D, Hoelzle K, Heinritzi K, Wittenb rin k MM. 2003. Development of a diagnostic PCR assay based on novel DNA sequences for the detection of M ycoplasma suis (Eperythrozoon suis) in porcine blood. Vet Microbiol 93: 185-196.

Hoelzle LE, Zeder M, Felder KM, Hoelzle K. 2014. Pathobiology of Mycoplasma suis. Vet J 202: 1, 20-25.

Kocher TD et al. 1989. Dynamics of mitochondrial DNA evolution in animals: amplification and sequencing with conserved primers. Proc Natl Acad Sci 86: 6, 196-200.

Kloster A et al. 1985. Eperythrozoonosis porcina: observaciones sobre la infección natural y experimental. Memorias VCong Arg Cs Vet Abs. p. 171.

Larkin Met al. 2007. Clustal Wand Clustal Xversion 2.0. Bioinformatics 23:

, 2947-2948. 14. Messick JB , Cooper SK, Huntley M. 1999. Development and evaluation of a polymerase chain reaction assay using the 16r RNA gene for detection of Eperythrozoon suis infection. J Vet Diagn Invest 11: 3, 229-236.

Messick J. 2004. Hemotrophic mycoplasmas (hemoplasmas): a review and new insights into pathogenic potential. Vet Clin Pathol 33: 1, 2-13.

Nonaka N, Thacker BJ, Schillhorn TW, Bull RW. 1996. In vitro maintenance of Eperythrozoon suis. Vet Parasitol 61: 3-4, 181-199.

Pereyra Net al. Prevalencia de la infección por el hemoplasma Mycoplasma suis en Argentina. Memorias del X IX Congreso Panamericano de Veterinaria.

Pintos ME. 2013. Estudio de las variaciones hematológicas, bioquímicas y de PCR, en cerdos esplenectomizados provenientes de una granja con antecedentes de Mycoplasma suis. X IV Jorn Divulg Técn Cient, Rosario, Abs. p. 144.

Pintos ME. 2016. Diagnóstico de Mycoplasma suis con técnicas convencionales y de biología molecular. Su relación con circovirus porcino tipo 2. Tesis Doctoral en Ciencias Veterinarias, FCV-UNLP, p. 56.

Rikihisa Y et al. 1997. Western immunoblot analysis of Haemobartonella muris and comparison of 16S rRNA gene sequences of H. muris, H. felis, and Eperythrozoon suis. J Clin Microbiol 35: 4, 823-829.

Schreiner SA et al. 2012. Nanotransformation of the haemotrophic Mycoplasma suis during in vitro cultivation attempts using modified cell free mycoplasma media. Vet Microbiol 160: 1, 227-232.

Seo MG, Kwon OD, Kwak D. 2019. Prevalence and phylogenetic analysis of hemoplasma species in domestic pigs in Korea. Parasit Vectors 12: 1, 378.

Stadler J et al. 2014. Clinical and haematological characterisation of Mycoplasma suis infections in splenectomised and non-splenectomised pigs. Vet Microbiol 172: 294-300.

Stadler J et al. 2019. Detection of Mycoplasma suis in presuckling piglets indicates a vertical transmission. BMC Vet Res 15: 252._

Strait EL, Hawkins PA, Wilson WD. 2012. Dysgalactia associated with Mycoplasma suis infection in a sow herd. J Am Vet Med Assoc 241: 12, 1666-1667.

Untergasser A et al. 2007. Primer3 Plus, an enhanced web interface to Primer3. Nucleic Acids Res 35: W71- W74.

Untergasser A et al. 2012. Primer3, new capabilities and interfaces. Nucleic Acids Res 40: 15, e115.

Watanabe Y, Fujihara M, Obara H, Nagai K, H arasawa R. 2011.Two genetic clusters in swine hemoplasmas revealed by analyses of the 16S rRNA and RNase P RNA genes. J Vet Med Sci 73: 12, 1657-1661.

Watanabe Y et al. 2012. Prevalence of swine hemoplasmas revealed by real-time PCR using 16S rRNA gene primers. J Vet Med Sci 74: 10, 1315-1318.

Weissenbacher LC et al. 2012. Porcine ear necrosis syndrome: a preliminary investigation of putative infectious agents in piglets and mycotoxins in feed. Vet J 194: 3, 392¬ 397.