Rahimabadi, E., Asadpour, Y., Pourbakhsh, S., Sayehban, P. (2017). Isolation and Identification of Mycoplasma agalactiae by Culture and Polymerase Chain Reaction Methods in the Sheep Herds in Guilan Province, Iran. Archives of Razi Institute, 72(4), 219-223. doi: 10.22092/ari.2017.113298
E. Rahimabadi; Y. Asadpour; S.A. Pourbakhsh; P. Sayehban. "Isolation and Identification of Mycoplasma agalactiae by Culture and Polymerase Chain Reaction Methods in the Sheep Herds in Guilan Province, Iran". Archives of Razi Institute, 72, 4, 2017, 219-223. doi: 10.22092/ari.2017.113298
Rahimabadi, E., Asadpour, Y., Pourbakhsh, S., Sayehban, P. (2017). 'Isolation and Identification of Mycoplasma agalactiae by Culture and Polymerase Chain Reaction Methods in the Sheep Herds in Guilan Province, Iran', Archives of Razi Institute, 72(4), pp. 219-223. doi: 10.22092/ari.2017.113298
Rahimabadi, E., Asadpour, Y., Pourbakhsh, S., Sayehban, P. Isolation and Identification of Mycoplasma agalactiae by Culture and Polymerase Chain Reaction Methods in the Sheep Herds in Guilan Province, Iran. Archives of Razi Institute, 2017; 72(4): 219-223. doi: 10.22092/ari.2017.113298
Isolation and Identification of Mycoplasma agalactiae by Culture and Polymerase Chain Reaction Methods in the Sheep Herds in Guilan Province, Iran
1Department of Veterinary, Guilan Agricultural and Natural Resources Research and Education Center, Rasht, Iran
2Mycoplasma Reference Laboratory, Razi Vaccine and Serum Research Institute, Agricultural Research, Education, and Extension Organization, Karaj, Iran
Abstract
Contagious agalactia is an infectious syndrome of sheep that is characterized by mastitis with reduction of milk production, arthritis, abortion, and keratoconjunctivitis. The disease is rapidly spread by the contact of the infected animals with the healthy ones. Domestic sheep and goats of both sexes can be infected at an equivalent frequency. Most of the researchers use culture and molecular methods for the isolation and identification of Mycoplasma. Mycoplasma agalactiae is the main cause of the disease in sheep. The aim of this study was to isolate and identify M. agalactiae by using culture and polymerase chain reaction (PCR) assay in the sheep herds in Guilan province, Iran. A total of 71 specimens were collected from seven sheep herds with clinical signs of agalactia disease. All of the seven sheep herds (100%) were positive either in PPLO agar or Mycoplasma PCR test. Out of the 71 specimens, 50 (70.4%) cases were positive; however, 21 (29.6%) samples were negative. Furthermore, 40 (80%) cases of the positive samples were detected for the presence of Mycoplasma by the PCR method; nonetheless, 34 (68%) samples were positive in culture. Additionally, out of the 40 positive samples in Mycoplasma PCR, 11 (27.5%) samples were detected in M. agalactiae-specific PCR. The samples that were positive for Mycoplasma were mostly detected in the ear/vaginal, milk, and ear swab samples, respectively, by culture and PCR methods. The most positive samples of Mycoplasma / M. agalactiae were obtained from the ear and vaginal samples. Our findings demonstrated that Mycoplasma was one of the main etiological agents of the contagious agalactia in Guilan province. In addition, PCR was found to be more successful than the culture method in the detection of Mycoplasma.
Isolement et identification de Mycoplasma agalactiae provenant des troupeaux de brebis de la province de Guilan en utilisant les méthodes de culture et de réaction en chaîne de la polymérase
Abstract [French]
Les infections à Mycoplasmes chez les petits ruminants dans les pays de la Méditerranée et les pays africains ont été identifiées ; ces infections engendrent de nombreux dégâts économiques. L'agalactie contagieuse est un syndrome infectieux du mouton qui est diagnostiqué par le gonflement du sein, l’interruption soudaine de la production de lait, l’enflure des articulations, l’avortement et l’inflammation de la cornée et de la conjonctive. Mycoplasma agalactiae est une cause majeure de la maladie chez les brebis. L’objectif de la présente étude est l’isolement et l’identification de Mycoplasma agalactiae provenant des troupeaux de brebis de la province de Guilan en utilisant les méthodes de culture et de réaction en chaîne de la polymérase. Pour ce faire, 71 échantillons provenant de 7 troupeaux de brebis avec des signes cliniques de la maladie d’Agalactie ont été recueillis. Tous les sept troupeaux (100%) échantillonnés étaient positifs à un des deux expériences de culture d'Agar ou la réaction en chaîne par polymérase. 50 échantillons sur 71 (70,4%) étaient positifs et 21 (29,6%) étaient négatifs. 40 échantillons positifs (80%) ont été détectés, en termes de Mycoplasma, dans le test de réaction en chaîne, mais 34 (68%) des échantillons étaient positifs en culture. 11 échantillons (27,5%) sur 40 échantillons identifiés de Mycoplasma, étaient de genre mycoplasma agalactiae. Les échantillons positifs de mycoplasma étaient plus fréquents dans les échantillons auriculaires et vaginaux, puis dans les échantillons d'oreille, du lait en culture et réaction en chaîne par polymérase. Mycoplasma / Mycoplasma Agalactiae étaient plus fréquemment détectés dans les échantillons provenant de l'oreille et du vagin. Mycoplasma agalactiae est l'une des principales causes d'Agalaxie contagieuse, qui a été identifiée pour la première fois dans les troupeaux de brebis dans la province de Guilan. Les résultats ont démontré que la méthode de la chaîne polymérase pourrait être une bonne alternative à la méthode de culture en raison d'un temps de diagnostic plus court.
Keywords [French]
Mycoplasma agalactiae, chaîne polymérase, Brebis, province de Guilan
References
Amores, J., Corrales, J.C., Martín, Á.G., Sánchez, A., Contreras, A., de la Fe, C., 2010. Comparison of culture and PCR to detect Mycoplasma agalactiae and Mycoplasma mycoides subsp. capri in ear swabs taken from goats. Veterinary Microbiol 140, 105-108.
Azevedo, E.O.d., Alcântara, M.D.B.d., Nascimento, E.R.d., Tabosa, I.M., Barreto, M.L., Almeida, J.F.d., et al., 2006. Contagious agalactia by Mycoplasma agalactiae in small ruminants in Brazil: first report. Brazilian Journal of Microbiology 37, 576-581.
Dedieu, L., Mady, V., Lefevre, P.-C., 1995. Development of two PCR assays for the identification of mycoplasmas causing contagious agalactia. FEMS microbiology letters 129, 243-249.
Greco, G., Corrente, M., Martella, V., Pratelli, A., Buonavoglia, D., 2001. A multiplex-PCR for the diagnosis of contagious agalactia of sheep and goats. Molecular and cellular Probes 15, 21-25.
Hajizadeh, A., Moazeni, J., Akhlagi, F., Naserirad, A., Motarez, B., Soleimani, S., et al., 2012. Isolation and identification of Mycoplasmas causing contagious
Agalactia syndrome in sheep & goats by using culture method and PCR in Ardebil province. Journal of Veterinary Laboratory Research 4, 271-281.
Kheirabadi, K., Ebrahimi, A., 2007. Investigation of Mycoplasma agalactiae in milk and conjunctival swab samples from sheep flocks in west central, Iran. Pakistan journal of biological sciences: PJBS 10, 1346-1348.
Kheirkhah, B., Pourbakhsh, S.A., Nadalian, M.-g., Banani, M., Ashtari, A., 2011. Detection of Mycoplasma agalactiae by culture and polymerase chain reaction (PCR) methods from Iranian goats. African Journal of Microbiology Research 5, 1668-1972.
Madanat, A., Zendulkova, D., Pospíšil, Z., 2001. Contagious agalactia of sheep and goats. A review. Acta Veterinaria Brno 70, 403-412.
Nicholas, R., Ayling, R., McAuliffe, L., 2008. Mycoplasma diseases of ruminants, CABI.
Ruffin, D.C., 2001. Mycoplasma infections in small ruminants. Veterinary Clinics of North America: Food Animal Practice 17, 315-332.
Subramaniam, S., Bergonier, D., Poumarat, F., Capaul, S., Schlatter, Y., Nicolet, J., et al., 1998. Species identification ofMycoplasma bovisandMycoplasma agalactiaebased on theuvrC genes by PCR. Molecular and cellular probes 12, 161-169.
Tola, S., Angioi, A., Rocchigiani, A., Idini, G., Manunta, D., Galleri, G., et al., 1997. Detection of Mycoplasma agalactiae in sheep milk samples by polymerase chain reaction. Veterinary microbiology 54, 17-22.
Zendulkova, D., Madanat, A., Lany, P., Rosenbergova, K., Pospíšil, Z., 2007. Detection of Mycoplasma agalactiae by Polymerase Chain Reaction in Jordanian sheep and goat herds. Acta Veterinaria Brno 76, 71-77.
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