Uso y desempeño de muestras de diferentes sitios del tracto respiratorio de los cerdos para la detección molecular y tipificación genética de Mycoplasma hyopneumoniae

  • Pablo Jesus Tamiozzo Departamento Patología Animal, Facultad de Agronomía y Veterinaria, Universidad Nacional de Río Cuarto. http://orcid.org/0000-0002-4501-9026
  • Mariné Tavella Departamento Patología Animal, Facultad de Agronomía y Veterinaria, Universidad Nacional de Río Cuarto.

Resumen

Mycoplasma hyopneumoniae es el agente causal de la Neumonía Enzoótica Porcina (NEP), una enfermedad que ocasiona grandes pérdidas económicas en las granjas porcinas de todo el mundo. Para el diagnóstico de la NEP, la presencia de lesiones pulmonares, de tos nos productiva, la detección de anticuerpos usando en enzimoinmunoensayo (ELISA) y la reacción en cadena de la polimerasa (PCR), son las herramientas más utilizadas.  Debido a la dificultad en el cultivo y aislamiento del agente, la PCR constituye una alternativa diagnóstica sensible, específica y rápida para determinar la infección causada por M. hyopneumoniae. Para la detección del agente, Varios formatos de PCR de primera y segunda generación a partir de diferentes muestras clínicas han sido descriptos en la literatura. Dado los abundantes antecedentes respecto a la detección del agente en algunas muestras clínicas más tradicionales, a lo novedoso del uso de otras muestras para tal fin y a la falta de información concisa que guíe a los veterinarios de campo, el objetivo del presente trabajo es realizar una reseña, resaltando los principales hallazgos, ventajas y desventajas, de los distintos tipos de muestras clínicas utilizadas para la detección molecular y el genotipado de M. hyopneumoniae.


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Citas

Abiven, P. & Pommier, P. (1993). Tracheobronchial lavage technique using the transnasal route for the detection of Mycoplasma hyopneumoniae in living non-anesthetized swine. Veterinary Research 24 (6):515-22.

Andrada, M. & Ambrogi, A. (1994). Estudio en matadero de pulmones de cerdos criados en sistemas al aire libre: Prevalencia y tipo de patologías. III Congreso Nacional de Producción Porcina. Rosario, Argentina 8 -10 Setiembre, 1994.

Baumeister, A., Runge, M., Ganter, M., Feenstra, A., Delbeck, F., Kirchhoff, H. (1998). Detection of Mycoplasma hyopneumoniaein Bronchoalveolar Lavage Fluids of Pigs by PCR. Journal of Clinical Microbiology. 36 (7): 1984–88.

Blanchard, B., Kobisch, M., Bové, J., Saillard, C. (1996). Polymerase chain reaction for Mycoplasma hyopneumoniae detection in tracheobronchiolar washings from pigs. Molecular and Cellular Probes. 10, 15–22.

Cai, Y., Van Dreumel, T., McEwen, B., Hornby, G., Bell-Rogers, P., Mc Raild, P., Josephson, G., Maxie, G. (2007). Application and field validation of a PCR assay for the detection of Mycoplasma hyopneumoniae from swine lung tissue samples. Journal of Veterinary Diagnostic Investigation.  19 (1): 91-5.

Calsamiglia, M., Pijoan, C., Trigo, A. (1999a). Application of a nested polymerase chain reaction assay to detect Mycoplasma hyopneumoniae from nasal swabs. Journal of Veterinary Diagnostic Investigation.  11(3):246-51.

Calsamiglia, M., Pijoan, C., Bosch, G. (1999b). Profiling Mycoplasma hyopneumoniae in farm using serology and a nested PCR technique. Journal of Swine Health and Production. 6: 263-8.

Calsamiglia, M., Collins, J.; Pijoan, C. (2000). Correlation between the presence of enzootic pneumonia lesions and detection of Mycoplasma hyopneumoniae in bronchial swabs by PCR. Veterinary Microbiology. 76: 299-303.

Caron, J., Ouardani, M, Dea, S. (2000). Diagnosis and differentiation of Mycoplasma hyopneumoniae and Mycoplasma hyorhinis infections in pigs by PCR amplification of the p36 and p46 genes. Journal of Clinical Microbiology. 38(4):1390-6.

Cheong, Y., Oh, C., L, Kee., Cho, K. (2017). Survey of porcine respiratory disease complex-associated pathogens among commercial pig farms in Korea via oral fluid method. Journal of Veterinary Science. 18 (3): 283-9.

Damte, D., Yohanes, S., Hossain, M., Lee, S., Rhee, M., KIM, Y., Park, S. (2014). Detection of naturally aerosolized Mycoplasma hyopneumoniae from the air of selected swine farms. Aerobiology 30: 205-9.

Dee, S., Otake, S., Oliveira, S., Deen, J. (2009). Evidence of long-distance airborne transport of porcine reproductive and respiratory syndrome virus and Mycoplasma hyopneumoniae. Veterinary Research. 40(4):39.

Dolso, I., Pelliza, B., Vissio, C., Carranza, A.,Ambrogi, A., Busso, J. Lesiones neumónicas halladas en matadero y su asociación con sistemas de crianza de cerdos al aire libre y confinados. Congreso MERCOSUR de Producción Porcina. Ciudad Autónoma de Buenos Aires, Argentina. 2000.

Dos santos, L., Sreevatsan, S., Torremorell, M., Moreira, M., Sibila, M., Pieters, M. (2015). Genotype distribution of Mycoplasma hyopneumoniae in swine herds from different geographical regions. Veterinary Microbiology. 175 (2-4):374-81. 

Dubosson, C., Conzelmann, C., Miserez, R., Boerlin, P., Frey, J., Zimmermann, W., Häni, H., Kuhnert, P. (2004). Development of two real-time PCR assays for the detection of Mycoplasma hyopneumoniaein clinical samples. Veterinary Microbiology. 102:55–65.

Fablet, C., Marois, C., Kobisch, M., Madec, F., Rose, N. (2010). Estimation of the sensitivity of four sampling methods for Mycoplasma hyopneumoniae detection in live pigs using a Bayesian approach. Veterinary Microbiology. 143(2-4):238-45.

Fano, E., Pijoan, C., Dee, S. (2005). Dynamics and persistence of Mycoplasma hyopneumoniae infection in pigs. Canadian Journal of Veterinary Research. 69(3): 223–8.

Fano, E., Pijoan, C., Dee, S., Deen, J. (2007). Effect of Mycoplasma hyopneumoniae colonization at weaning on disease severity in growing pigs. Canadian Journaal of Veterinary Research, 71(3):195-200.

Fourour, S., Fablet, C., Tocqueville, V., Dorenlor, V., Eono, F., Eveno, E., Kempf, I., Marois-Créhan, C. (2018). A New Multiplex Real-Time TaqMan ® PCR for Quantification of Mycoplasma hyopneumoniae, M. hyorhinis and M. flocculare: Exploratory
Epidemiological Investigations to Research Mycoplasmal Association in Enzootic Pneumonia-Like Lesions in Slaughtered Pigs. Journal of Applied Microbiology. 125 (2): 345-55.

Garza Moreno, L., Segalés, J., Aragón, V., Correa-Fiz, F., Pieters, M., Carmona, M., Krejci, R., Sibila, M. (2019). Characterization of Mycoplasma hyopneumoniae strains in vaccinated and non-vaccinated pigs from Spanish slaughterhouses. Veterinary Microbiology. 231:18-23.

Giacomini, E., Ferrari, N., Pitozzi, A., Remistani, M., Giardiello, D., Maes, D., Alborali, G. (2016). Dynamics of Mycoplasma hyopneumoniae seroconversion and infection in pigs in the three main production systems. Veterinary Research Communications. 40 (2):81-8.

Harasawa, R., Koshimizu, K., Takeda, O., Uemori, T., Asada, K. Y Kato, I. (1991). Detection of Mycoplasma hyopneumoniae DNA by the Polymerase Chain Reaction. Mol Cell Probes 5 (2), 103-9.

Hermann, J., Brockmeier, S., Yoon, K., Zimmermann, J. (2008). Detection of respiratory pathogens in air samples from acutely infected pigs. Canadian Journal of Veterinary Research. 72(4):367-70.

Hernandez-Garcia, J., Robben, N., Magnée, D., Dennis, I., Kayes, S., Thomson, J., Tucker, A. (2017). The use of oral fluids to monitor key pathogens in porcine respiratory disease complex. Porcine Health Management. 3:7

Johansson, K., Mattsson, J., Jacobsson, K., Fernandez, C., Bergström, K., Bölske, G., Wallgren, P., Göbel, U. (1992). Specificity of oligonucleotide probes complementary to evolutionarily variable regions of 16S rRNA from Mycoplasma hyopneumoniaeand
Mycoplasma hyorhinis. Research in Veterinary Science 52: 195-204.

Kuhnert, P., Overesch, G., Belloy, L. (2011). Genotyping of Mycoplasma hyopneumoniaein wild boar lung samples. Veterinary Microbiology 152: 191–5.

Kuhnert, P. & Overesch, G. (2014). Molecular epidemiology of Mycoplasma hyopneumoniae from outbreaks of enzootic pneumonia in domestic pig and the role of wild boar. Veterinary Microbiology. 174 (1-2): 261-6. 

Kurth, K., Hsu, T., Snook, E., Thacker, B., Minion, F. (2002). Use of a Mycoplasma hyopneumoniae nested polymerase chain reaction test to determine the optimal sampling sites in swine. Journal of Veterinary Diagnostic Investigation 14(6):463-9.

Makhanon, M., Tummaruk, P., Thongkamkoon, P., Thanawongnuwech,R., Prapasarakul, N. (2012). Comparison of detection procedures of Mycoplasma hyopneumoniae, Mycoplasma hyosynoviae, and Mycoplasma hyorhinis in lungs, tonsils, and
synovial fluid of slaughtered pigs and their distributions in Thailand. Tropical Animal Health and Production 44: 313–18.

Marois, C., Dory, D., Fablet, C., Madec, F., Kobisch, M. (2010). Development of a quantitative Real-Time TaqMan PCR assay for determination of the minimal dose of Mycoplasma hyopneumoniaestrain 116 required to induce pneumonia in SPF pigs. Journal of Applied Microbiology 108 (5): 1523-33. 

Mattsson, J., Bergström, K., Wallgren, P., Johansson, K. (1995). Detection of Mycoplasma hyopneumoniae in Nose Swabs from Pigs by In Vitro Amplification of the 16S rRNA Gene. Journal of Clinical Microbiology 893–897.

Mayor, D., Zeeh, F., Frey, J.; Kuhnert, P. (2007). Diversity of Mycoplasma hyopneumoniaein pig farms revealed by direct molecular typingof clinical material. Veterinary Research (38): 391–8.

Mayor, D., Jores, J., Korczak, B., Kuhnert, P. (2008). Multilocus sequence typing (MLST) of Mycoplasma hyopneumoniae: A diverse pathogen with limited clonality. Veterinary Microbiology 127: 63–72.

Meyns, T., Maes, D., Dewulf, J., Vicca, J., Haesebrouck, F., Kruif, A. (2004). Quantification of the spread of Mycoplasma hyopneumoniae in nursery pigs using transmission experiments. Preventive Veterinary Medicine (66): 265–75.

Meyns, T., Dewulf, J., Kruif, A., Calus, D., Haesebrouck, F., Maes, D. (2006). Comparison of transmission of Mycoplasma hyopneumoniaein vaccinated and non-vaccinated populations. Vaccine (24):7081–6.

Michiels, A., Vranckx, K., Piepers, S., Del Pozo Sacristan, R., Arsenakis, I., Boyen, F., Haesebrouck, F., Maes, D. (2017). Impact of diversity of Mycoplasma hyopneumoniae strains on lung lesions in slaughter pigs. Veterinary Research. 48(1):2.

Moiso, N., Degano, F., Pieters, M., Camacho, P., Estanguet, A., Parada, J., Tamiozzo, P. (2020). Detection of Mycoplasma hyopneumoniae in nasal and laryngeal swab specimens in endemically infected herds. The Veterinary Record 186 (1):27.

Otagiri, Y., Asai, T, Okada, M., Uto, T., Yazawa, S., Hirai, H., Shibata, I., Sato, S. (2005). Detection of Mycoplasma hyopneumoniae in lung and nasal swab samples from pigs by nested PCR and culture methods. Journal of Veterinary Medicine Science. 67(8): 801-5.

Otake, S., Dee, S., Corso, C., Oliviera, S., Deen, J. (2010). Long-distance airborne transport of infectious PRRSV and Mycoplasma hyopneumoniae from a swine population infected with multiple viral variants. Veterinary Microbiology 145 (3-4):198-208.

Overesch, G. & Kuhnert, P. (2017). Persistence of Mycoplasma hyopneumoniae sequence types in spite of a control program for enzootic pneumonia in pigs. Preventive Veterinary Medicine. 145:67-72. 

Pantoja, L., Pettit, K., Dos Santos, L., Tubbs, R., Pieters, M. (2016). Mycoplasma hyopneumoniaegenetic variability within a swine operation. Journal of Veterinary Diagnostic Investigation 28 (2): 175–9.

Pelliza, B., Miguez, M., Parsi, J. Impacto productivo de Mycoplasma hyopneumoniae en cerdos criados en sistemas al aire libre. VI Congreso Nacional de Producción Porcina. Rosario, Argentina 5-7 noviembre 1998.

Perfumo, C., Sanguinetti, R., Risso, M., Aguirre, J., Armocida A. Prevalencia en frigorífico de lesiones compatibles con Neumonía Enzoótica Porcina en animales provenientes de establecimientos de cría intensivo. III Congreso Nacional de Producción
Porcina. Rosario, Argentina 8 -10 Setiembre, 1994.

Pieters, M., Pijoan, C., Fano, E.; Dee, S. (2009). An assessment of the duration of Mycoplasma hyopneumoniae infection in an experimentally infected population of pigs. Veterinary Microbiology (134) 261–6.

Pieters, M., Daniels, J., Rovira, A. (2017). Comparison of sample types and diagnostic methods for in vivo detection of Mycoplasma hyopneumoniae during early stages of infection. Veterinary Microbiology (203) :103-9.

Rebaque, F., Camacho, P., Parada, J., Lucchesi, P., Ambrogi, A., Tamiozzo, P. (2018). Persistence of the same genetic type of Mycoplasma hyopneumoniae in a closed herd for at least two years. Revista Argentina de Microbiología 50 (2):147-50. 

Roos, L., Fano, E., Homwong, N., Payne, B., Pieters, M. (2016). A model to investigate the optimal seeder-to-naïve ratio for successful natural Mycoplasma hyopneumoniae gilt exposure prior to entering the breeding herd. Veterinary Microbiology 184:51-8.

Ruiz, A., Galina, L., Pijoan, C. (2002). Mycoplasma hyopneumoniae colonization of pigs sired by different boars. Canadian Journal of Veterinary Research 66 (2):79-85.

Savic, B., Ivetic, V., Milicevic, V., Pavlovic, I., Zutic, M., Gagrcin, M. (2010). Genetic diversity of Mycoplasma hyopneumoniae isolates from conventional farrow-to-finish pig farms in Serbia. Acta Veterinaria Hungarica. 58(3): 297-308. 

Sibila, M., Calsamiglia, M., Segalés, J., Rosell, C. (2004a). Association between Mycoplasma hyopneumoniae at different respiratory sites and presence of histopathological lung lesions. The Veterinary Record 155(2):57-8.

Sibila, M., Calsamiglia, M., Vidal, D., Badiella, L., Aldaz, A., Jensen, J. (2004b). Dynamics of Mycoplasma hyopneumoniae infection in 12 farms with different production systems. Canadian Journal of Veterinary Research.  68:12-8.

Sibila, M., Nofrarías, M., Lopez-Soria, S., Segalés, J., Riera, P., Llopart, D., Calsamiglia, M. (2007a). Exploratory Field Study on Mycoplasma hyopneumoniae Infection in Suckling Pigs. Veterinary Microbiology 121:352-6.

Sibila, M.; Nofrarías, M.; Lopez-Soria, S.; Segalés, J.; Valero, O.; Espinal, A.; Calsamiglia, M. (2007b). Chronological study of Mycoplasma hyopneumoniae infection, seroconversion and associated lung lesions in vaccinated and non-vaccinated pigs.
Veterinary Microbiology 122: 97–107.

Sibila, M., Bernal, R., Torrents, D., Riera, P., Llopart, D., Calsamiglia, M., Segalés, J. (2008). Effect of sow vaccination against Mycoplasma hyopneumoniae on sow and piglet colonization and seroconversion, and pig lung lesions at slaughter. Veterinary Microbiology 127: 165–170.

Sosa, C., Blois, A., Ibáñez, F., Tamiozzo, P. (2019). Genetic diversity of Mycoplasma hyopneumoniae in Mendoza province. Revista Argentina de Microbiología. 51(3):229-33.

Sponheim, A., Álvarez, J., Fano, E., Schmaling, E., Dee, S., Hanson, D., et al…(2020). Comparison of the Sensitivity of Laryngeal Swabs and Deep Tracheal Catheters for Detection of Mycoplasma hyopneumoniae in Experimentally and Naturally Infected
Pigs Early and Late After Infection. Veterinary Microbiology  241:108500. 

Stakenborg, T., Vicca, J., Maes, D. (2006). Comparison of molecular techniques for the typing of Mycoplasma hyopneumoniaeisolates. Journal of Microbiology Methods. 66 (2): 263-275.

Stärk, K., Nicolet, J., Frey, J. (1998). Detection of Mycoplasma hyopneumoniae by air sampling with a nested PCR assay. Applied Environmental Microbiology 64 (2):543-8.

Strait, E., Madsen, M., Minion, F., Christopher-Hennings, J., Dammen, M., Jones, K., Thacker, E. (2008). Real-Time PCR Assays to Address Genetic Diversity among Strains of Mycoplasma hyopneumoniae.. Journal Of Clinical Microbiology 46 (8): 2491–8.

Takeuti, K., De Barcellos, D., De Andrade, C., De Almeida, L., Pieters, M. (2017a). Infection dynamics and genetic variability of Mycoplasma hyopneumoniae in self-replacement gilts. Veterinary Microbiology (208) 18–24.

Takeuti, K., De Barcellos, D., De Lara, A., Kunrath, C., Pieters, M. (2017b). Detection of Mycoplasma hyopneumoniae in naturally infected gilts over time. Veterinary Microbiology 203: 215–220.

Takeuti, K., De Barcellos, D., Pieters, M. (2017c). Mycoplasma hyopneumoniae detection in nylon-flocked and rayon-bud swabs. Journal of Microbiological Methods 141: 118-20.

Tamiozzo, P., Pelliza, B., Carranza, A., Ambrogi, A. (2011a). Monitoramento da presença de Mycoplasma hyopneumoniae em granjas de suínos durante a implementação de programas de erradicação. Ciência Rural 41 (4): 699-705.

Tamiozzo, P, Carranza, A, Parada, J, Pelliza, B, Ambrogi, A. (2011b). Algunos aspectos sobre el estado de colonización y estado serológico en cerdas vacunadas contra Mycoplasma hyopneumoniae y de sus lechones según el número ordinal de partos. Archivos de Medicina Veterinaria, 43 (3): 251-8.

Tamiozzo, P., Lucchesi, P., Ambrogi, A. 2011c. Genetic diversity of Mycoplasma hyopneumoniae in pig farms from Argentina. InVet 13 (1): 27-35.

Tamiozzo P, Zamora R, Lucchesi PM, Estanguet A, Parada J, Carranza A, Camacho P, Ambrogi A. (2015). MLVA typing of Mycoplasma hyopneumoniae bacterins and field strains. The Veterinary Record Open, 2 (2): e000117.

Vangroenweghe, F., Karriker, L., Main, R., Christianson, E., Marsteller, T., Hammen, K., et al …2015. Assessment of litter prevalence of Mycoplasma hyopneumoniae in preweaned piglets utilizing an antemortem tracheobronchial mucus collection technique and a real-time polymerase chain reaction assay. Journal of Veterinary Diagnostic Investigation 27 (5):606-10.

Vangroenweghe, F., Willems, E., Malášek, J., Thas, O., Maes, D. (2018a). Use of trachea-bronchial swab qPCR testing to confirm Mycoplasma hyopneumoniae seropositivity in an SPF breeding herd. Porcine Health Management 4:12.

Vangroenweghe, F., Willems, E., Thas, O., Maes, D. (2018b). Confirmation of Mycoplasma hyopneumoniaein a breeding herd through tracheobronchial swab sampling and PCR. The Veterinary Record 183 (10): 325.

Verdin, E., Saillard, C., Labbé, A., Bové, J., Kobisch, M. (2000a). A nested PCR assay for the detection of Mycoplasma hyopneumoniae in tracheobronchiolar washings from pigs. Veterinary Microbiology 76: 31-40.

Verdin, E., Kobisch, M., Bové, J., Garnier, M., Saillard, C. (2000b). Use of an internal control in a nested-PCR assay for Mycoplasma hyopneumoniaedetection and quantification in tracheobronchiolar washings from pigs. Molecular and Cellular Probes 14: 365–72.

Vilalta, C., Sanhueza, J., Murray, D., Johnson, L., Pieters, M. (2019). Detection of Mycoplasma hyopneumoniae in piglet processing fluids. The Veterinary Record 185 (16): 510.

Vranckx, K., Maes, D., Calus, D., Villarreal, I.., Pasmans, F., Haesebrouck, F. (2011). Multiple-Locus Variable-Number Tandem-Repeat Analysis Is a Suitable Tool for Differentiation of Mycoplasma hyopneumoniae Strains without Cultivation. Journal Of Clinical Microbiology 49 (5):2020–3.

Vranckx, K., Maes, D., Marchioro, S., Villarreal, I., Chiers, K., Pasmans, F., Haesebrouck, F. (2012). Vaccination reduces macrophage infiltration in bronchus-associated lymphoid tissue in pigs infected with a highly virulent Mycoplasma hyopneumoniae strain. BMC Veterinary Research 8:24.

Wolley, L., Fell, S., Gonsalves, J., Walker, M., Djordjevic, S., Jenkins, C., Eamens, G. (2012). Evaluation of clinical, histological and immunological changes and qPCR detection of Mycoplasma hyopneumoniae in tissues during the early stages of mycoplasmal pneumonia in pigs after experimental challenge with two field isolates. Veterinary Microbiology 161: 186–95.

Zeeh, F., Kuhnert, P., Miserez, R., Doherr, MG., Zimmermann, W. (2005). Field validation of a real-time PCR test for the detection of Mycoplasma hyopneumoniae in nasal swabs of live pigs. Schweiz Arch Tierheilkd. 147(9):373-9.

Zielinski, G. & Ross, R F. (1992). Morphologic Features and Hydrophobicity of the Cell Surface of Mycoplasma hyopneumoniae. American Journal of Veterinary Research 53 (7): 1119-24.

Zielinski, G. Evaluación de parámetros productivos en cerdos criados al aire libre vacunados contra neumonía enzoótica. V Congreso Nacional de Producción Porcina. Río Cuarto, Argentina 5- 8 octubre 1996.
Publicado
2021-08-06
##submission.howToCite##
TAMIOZZO, Pablo Jesus; TAVELLA, Mariné. Uso y desempeño de muestras de diferentes sitios del tracto respiratorio de los cerdos para la detección molecular y tipificación genética de Mycoplasma hyopneumoniae. Ab Intus, [S.l.], n. 7, p. 60-72, aug. 2021. ISSN 2618-2734. Disponible en: <http://www.ayv.unrc.edu.ar/ojs/index.php/Ab_Intus/article/view/165>. Fecha de acceso: 21 sep. 2021
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