Kemal METİNER, Serkan İKİZ, Seyyal AK, Atilla ILGAZ, Funda BAĞCIGİL, N. Yakut ÖZGÜR

Domuzlarda Actinobacillus pleuropneumoniae Varlığının PCR ile Araştırılması

Türkiye’nin farklı bölgelerinde yetiştirilerek kesime gönderilen 1 ay - 3 yaş arasındaki 312 domuz’a ait tonsil örneğinde Actinobacillus pleuropneumoniae spesifik DNA’sı, dsbE like geninden derive primerler (5’GATAAACCTTTTCCGGAATT-3’ ve 5’-TACCACACCGTGTTTATCAA-3’) kullanılarak PCR ile araştırıldı. PCR sonucunda tonsil örneklerinin hiç birinde 342 bp’lik bantlar görülmedi. Örneklerin PCR negatif olarak saptanması, domuz pleuropnömonisinin incelenen işletmelerde bulunmadığını ortaya koyması yönünden sevindiricidir. Bu ve diğer infeksiyöz hastalıklar açısından yeni çalışmaların yapılması gerekmektedir.

INVESTIGATION OF THE PRESENCE OF ACTINOBACILLUS PLEUROPNEUMONIAE BY PCR IN PIGS

A. pleuropneumoniae specific DNA in tonsil samples of 1 month to 3 years old pigs raised in different regions and send to slaughterhouses was investigated by PCR using primers (5-’GATAAACCTTTTCCGGAATT-3’ and 5’- TACCACACCGTGTTTATCAA-3’) derived from dsbE like gen. In neither tonsil sample, 342 bp bands were observed by PCR. Determining the samples as PCR negative is promising that the infection is not present in the investigated farms and further studies should be performed for this and other possible infectious diseases of pigs.

PDF

___

Chevallier, B., Blanchard, B., Keita, A., Pagot, E., Pommier, P., 2 0 0 0 . Detection of Actinobacillus pleuropneumoniae by PCR: field evaluation. Journées Rech. Porcine en France. 3 2 , 1 - 6 .
Chiers, K., Done, E., Overbeke, I.V., Ducatelle, R., Haesebouck, F., 2002a. Actinobacillus pleuropneumoniae infections in closed swine herds: infection patterns and serological profiles. Veterinary Microbiology 85,343-352.
Chiers, K., Done, E., Overbeke, I.V., Ducatelle, R., Haesebouck, F., 2002b. Evaluation of serology, bacteriological isolation and polymerase chain reaction for the detection of pigs carrying Actinobacillus pleuropneumoniae in the upper respiratory tract after experimental infection. Veterinary Microbiology 88, 385- 392.
Chiers, K., Overbeke, I.V., Done, E., Baele, M., Ducatella, R., Beare, De T., Haesebouck, F., 2001. Detection of Actinobacillus pleuropneumoniae in cultures from nasal and tonsillar swabs of pigs by a PCR assay based on the nucleotide sequence of a dsbE-like gene. Veterinary Microbiology 83, 147-159.
Da Costa ,M.M., Klein, S.C.,Balestrin, R., Shrank, A., Piffer I.A., da Silva, S.C., Schrank I.S., 2004. Evaluation of PCR based on gene apxIVA associated with 16S rDNA sequencing for the identification of Actinobacillus pleuropneumoniae and related species. Current Mikrobiology. 48, 189-195
Dubreuil, J.D., Jacques, M., Mittal, K.R. and Gottschalk, M., 2000. Actinobacillus pleuropneumoniae surface polysaccarides; their role in diagnosis and immunogenicity. Animal Health Research Reviews 1, 73-93.
Gottschalk, M., Broes, A., Mittal, K.R., Kobisch, M., Kuhnert, P. Lebrun, A., Frey, J., 2003. Non-Pathogenic Actinobacillus isolates antigenically and biochemically similar to Actinobacillus pleuropneumoniae: a novel species? Veterinary Microbiology 92, 87-101.
Gram, T., Ahrens, P., Nielsen, J.P., 1996. Evaluation of PCR for detection of Actinobacillus pleuropneumoniae in mixed bacterial cultures from tonsils. Veterinary Microbiology 51, 95- 104.
Holt, J.G., Krieg, N.R., Sneath, P.H.A., Staley, J.T., Williams, S.T., 1994. Bergey's Manual of Determinative Bacteriology. 9. ed. Williams & Wilkins, Baltimore USA, 194 -196, 275 – 277. 22
İnzana, T.J., Jianneng, M.A., Workman T., Gogolewski, R.P. and Anderson, P., 1988. Virulence properties and protective efficacy of the capsular polymer of Haemophilus (Actinobacillus) pleuropneumoniae serotype 5. Infection and Immunity 56,1880-1889.
Levonen, K., 2001. The dedection of respiratory diseases in swine herds by means of antibody assay on collostrum from sows. Academic Dissertation, Faculty of Veterinary Medicine, University of Helsinki, Hesinki.
MacInnes, J.I., Rosendal, S., 1988. Prevention and kontrolof Actinobacillus (Haemophilus) pleuropneumoniae infection in swine: a reviw. The Canadian Veterinary Journal 29, 572-574.
Metiner, K., Ak, S., 2007. Presence and seroprevalence of Actinobacillus pleuropneumoniae in pigs in Turkey. Acta Veterinaria Brno 76, 237-244.
Moral, C.H., Sosano, C., Salazar, M.S., Marcos, J.Y., Ramos, S.S., and Carrasco, G.N., 1999. Molecular cloning and sequencing of the aroA gene from Actinobacillus pleuropneumoniae and its use in a PCR assay for rapid identification. Journal of Clinical Microbiology , 37, 1575-1578.
Rycroft, A.N. and Garside, L.H., 2000. Actinobacillus species and their role in animal disease. The Veterinary Journal 159, 18-36.
Schaller, A., Djordjeviç, P.S., Eamenens, J.G., Forbes, A.W., Kuhn, R., Kuhnert, P., Gottschalk, M., Nicolet, J., Frey, J., 2001. Identification and detection of Actinobacillus pleuropneumoniae by PCR based on the gene apxIVA. Veterinary Microbiology 79, 47-62.
Sidibe, M., Messier, S., Lariviere, S., Gottschalk, M. and Mittal, K.R., 1993. Detection of Actinobacillus pleuropneumoniae in the porcine upper respiratory tract as a complement to serological tests.Canadian Journal of Veterinary Research 57, 204-208.
Tonpitak, W., Rohde, J., Gerlach, G.F, 2007. Prevalence of ‘‘Actinobacillus porcitonsillarum’’ in porcine tonsils and development of a diagnosis duplex PCR differentiating between ‘‘Actinobacillus porcitonsillarum’’ and Actinobacillus pleuropneumoniae. Veterinary Mikrobiology 122,157-165
TÜİK, 2010. Türkiye hayvansal üretim istatistikleri http://www.tuik.gov.tr (Erişim 10.11.2010)
XIAO Guo-sheng, CAO San- Jie, DUAN Lİ-li, WEN Xin-tian, MaXiao-ping and CHEN Hua mei., 2006. Identification and detection of Actinobacillus pleuropneumoniae in infected and subclinical infected pigs by multiplex PCR based on the genes ApxIVA and OmlA. Agricultural Sciences in China 5, 146-154