Identifikasi Molekuler Pasteurella multocida Penyebab Pasteurellosis Pada Babi Di Yogyakarta
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Victor Lenda(1*)
Program Studi Kesehatan Hewan Jurusan Peternakan Politeknik Pertanian Negeri Kupang
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Hermilinda Parera(2)
Program Studi Kesehatan Hewan Jurusan Peternakan Politeknik Pertanian Negeri Kupang
(*) Corresponding Author
Abstract
The purpose of this study was to determine the genetic variation of P.multocida in Yogyakarta with other overseas P. multocida strains. Samples were obtained from necropted intensive pig farming with clinically pneumonia symptoms, specific pulmonary lessions and then followed by histopathological examination and b iochemical characterization of the isolates. Then followed by histopathological examination and biochemical characterization of the isolates. Molecular
test performed by DNA extraction using the QIAGEN QIAamp DNA minikit, amplification of the 16S rRNA gene using forward primer 5' GGA GTG AAC TGC AGC TAA TAC C 3', and reverse primer 5' GTA GGT AAG CTT CGC GTT GTT G 3', electrophoresis, purification and sequencing. Macroscopic and histopathological examination results were analyzed descriptively. Sequencing r esults were analyzed by multiple alignment with other Pasteurella spp. taken from GenBank using the Clustal W software, subsequently analyzed using Neighbour Joining and Maximum Parsimony method that exist in program MEGA version 5.1 The results showed gen etic distance based on 765 nucleotides of 16S rRNA gene of P. multocida isolates from lungs of bronchopneumonia of swine in Yogyakarta at 0%. Filogram based on the nucleotide sequence showed a high similarity between P. multocida isolates from Yogyakarta a nd other isolates from USA, Germany, China, Europe, and Hungaria.
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References
Cameron, R.D., O’Boyle, D., Frost A.J., Gordon A.N., and Fegan, N., 1996. An outbreak of haemorrhagic septicaemia associated with Pasteurella multocida subsp gallicida in large pig herd. Aust. Vet. J., 73:27–29.
Davies, R. L., MacCorquodale, R., Baillie, S. and Caffrey B., 2003. Characterization and comparison of Pasteurella multocida strains associated with porcine pneumonia and atrophic rhinitis. J. Med. Microbiol. 52:59–67.
Davies, R. L., 2004. Genetic diversity among Pasteurella multocida strains of avian, bovine, ovine and porcine origin from England and Wales by comparative sequence analysis of the 16S rRNA gen. Microbiology. 150: 4199–421.
Dey, S., Singh, V.P., Kumar, A.A., Sharma, B., Srivastava, S.K., Singh, N., 2007. Comparative sequence analysis of 16S rRNAgene of Pasteurella multocida serogroup B isolates from different animal spesies. Veterinary Science 83:1-4.
Higgins D., Thompson J., Gibson T., Thompson J. D., Higgins D. G., Gibson T. J., 1994. CLUSTAL W: improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choice. Nucleic Acids Res. 22:4673-4680.
Jamaludin, R., Blackall, P.J., Hansen, M.F., Humphrey, S., Styles, M., 2005. Phenotypic and genotypic characterisation of Pasteurella multocida isolated from pigs at slaughter in New Zealand. N. Z. Vet. J. 53, 203-20.
Kumar, H., Mahajan, V., Sharma, S., Alka, Singh R., Arora, A. K., Banga, H. S., Verma, S., Kaur, K., Kaur, P., Meenakshi, Sandhu, K. S., 2007. Concurrent pasteurellosis and classical swine fever in Indian pigs. Journal of Swine Health and Production. 15(5):279-283.
Lopez, A. 2001. Respiratory System, Thoracic Cavity and Pleura. In: Thomson’s Special Veterinary Pathology, 3rd Ed. McGavin, M. D., Carlton W. W. & Zachary, J., (eds.), Mosby-Year Book Inc., Pp. 125-195.
Mackie, J.T., Barton M., and Kettlewell, J., 1992. Pasteurella multocida septicaemia in pigs. Aust. Vet. J. (69) Pp. 227–228.
MacFaddin, J. F. 1980. Biochemical Tests for Identification of Medical Bacteria, 2nd ed. The Williams & Wilkins Co.,Baltimore. p. 527.
Pijoan, C., 2006. Pneumonic pasteurellosis. In: Diseases of Swine, 9th ed. (Eds.) Straw B. et al. Ames, IA: Iowa State University Press. Blackwell Publishing Australia. Pp. 719-725.
Pors, S. E., Hansen M. S. , Bisgaard, M., , Jensen, H. E. , 2011. Occurrence and associatedlesions of Pasteurella multocida in porcine bronchopneumonia. Vet. Microbiol. 150 (1-2):160-166.
Quinn, P.J.,Markey, B.K., Leonard, F. C., Hartigan, P., Fanning, S., Fitzpatrick, E.S., 2003. Veterinary Microbiology and Microbial Disease. 2nd Ed. John Wiley & Sons, Iowa. Pp. 137-143.
Saitou, N. and Nei, M., 1987. The neighbor-joining method: A new method for reconstructing phylogenetic trees. Molecular Biology and Evolutio. 4:406-425.
Tamura, K., Peterson, D., Peterson, N., Stecher, G., Nei, M., and Kumar, S., 2011. MEGA5: Molecular Evolutionary Genetics Analysis using Maximum Likelihood, Evolutionary Distance, and Maximum Parsimony Methods. Molecular Biology and Evolution. 28:2731-2739.
Townsend, K.M., Frost, A.J., Lee, C.W., Papadimitriou, J.M., Dawkins, H.J., 1998a. Development of PCR assays for species- and type-specific identification of Pasteurella multocida isolates. J. Clin. Microbiol. 36: 1096-1100.
Townsend, K.M., O'Boyle, D., Phan, T.T., Hanh, T.X., Wijewardana, T.G., Wilkie, I., Trung, N.T., Frost, A.J., 1998b. Acute septicaemic pasteurellosis in Vietnamese pigs. Vet. Microbiol. 63:205-215.
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