Study of BuLA-DRB3 polymorphism in Khuzestan river buffaloes

Authors

1 Department of Microbiology and Immunology, Faculty of Veterinary Medicine, University of Tehran, Tehran-Iran

2 Department of Clinical Sciences, Faculty of Veterinary Medicine, University of Chamran, Ahvaz-Iran

3 Faculty of veterinary medicine, Shahid Chamran University of Ahvaz, Ahvaz, Iran

4 Graduated PhD student at Department of Microbiology and Immunology, Faculty of Veterinary Medicine, University of Tehran, Tehran-Iran

Abstract

BACKGROUND: Major histocompatibility complex (MHC) comprises a group of genes, which plays a central role in immune response. The exon 2 of BuLA-DRB3 is part of the MHC class II in buffalo that highly polymorphic, found to be associated with resistance/susceptibility to infections and also with production parameters. OBJECTIVES: The purpose of the present study is to identify BuLA-DRB3 polymorphism in Khuzestan buffaloes and compare this population with other Iranian and world buffalo populations. METHODS: Blood samples were taken from 136 unrelated Khuzestan river buffaloes. After DNA extraction, second exon of BuLA-DRB3 was amplified by the seminested PCR method. Then, the fragments produced by amplifying second exon were cut by RsaI restriction enzyme according to van Eijik method. In the following, allelic frequencies, genotype frequencies, expected and observed homozygosty and heterozygosity were calculated. RESULTS: In restriction fragment analysis 13 and 24 different allelic and genotype patterns were identified for RsaI restriction enzyme, respectively. 10 out of 13 alleles were previously reported. The most frequent genotype was oo)0.1691) and then followed by hh (0.1544) ,ll(0.1103), lw (0.0955), lh (0,0808), ha (0.0661) and lo (0.0514). Also four most frequent alleles were o (0.2721), h (0.2316),l (0.2316)and w (0.1176), respectively. These seven genotypes and four alleles form 72.76% and74.29% overall genotype and allele frequency of population.  In addition, estimation of heterozygosity/homozygosty and deviation from Hardy-Weinberg equilibriumofcorresponding population revealed observed homozygosty is more than heterozygosity and departure of population from Hardy-Weinberg equilibrium. CONCLUSIONS: The results indicated that exon 2 of the BuLA-DRB3 gene is highly polymorphic among Khuzestan buffaloes and although, there is differences between buffalo’s genetic polymorphism of distinct world regions, Khuzestan buffaloes’ population is similar to Egyptian buffaloes’ population.

Keywords


Acharya, C.P., Pipalia, D.L., Rank, D.N., Joshi, C.G., Brahmkshtri, B.P., Solanki, J.V., Shah, R.R. (2002) BOLA-DRB3 gene polymorphism in Jafarabadi and Mehsani buffaloes as revealed by PCR-RFLP. Ind  Vet J. 79: 652-656.##
Behl, J.D., Verma, N.K., Behl, R., Mukesh, M., Ahlawat, S.P.S. (2007) Characterization of genetic polymorphism of the bovine lymphocyte antigen DRB3.2 locus in Kankrej cattle (Bos indicus). J Dairy Sci. 90: 2997-3001.##
De, S., Singh, R.K., Butchaiah, G. (2002) MHC-DRB exon 2 allele polymorphism in Indian river buffalo (Bubalus bubalis). Anim Genet. 33: 215-219.##
Kumar, S., Sangwan, M.L., Rupender, A. (2008) Polymorphismin DRB3 exon 2 by PCR-RFLP and its association with mastitis in Nili-Ravi breed. Indian J Biotech. 7: 398-400.##
Kumar, S., Sangwan, M.L., Ahlawat, S., Barwar, A. (2011) Polymorphismin DRB3 exon 2 by PCR-RFLP and its association with mastitis in Murrah buffaloes. Indian J Biotech. 10: 232-234.##
Levene, H. (1949) On a matching problem arising in genetics. Ann Math Stat. 20: 91-94.##
Mosafer, J., .Heydarpour, M., Manshad, E., Russell, G., Sulimova, G.E. (2012) Distribution of BoLA-DRB3 Allelic Frequencies and Identification of Two New Alleles in Iranian Buffalo Breed. Sci World J. 12: 22-28.##
Nanda, A.S., Nakao, T. (2003) Role of buffalo in the socioeconomic development of rural Asia: Current status and future prospectus. Anim Sci J. 74: 443-455.##
Nei, M. (1973) Analysis of gene diversity in subdivided populations. Proc Natl Acad Sci. 70: 3321-3323.##
Othman, O.E., Ahmed, S.S. (2010) Genetic polymorphism of BoLA-DRB3 exon 2 in egyptian buffalo. Genes Genomes Genomics. 4: 70-73.##
Pasha, T.N., Hayat, Z. (2012) Present situation and future perspective of buffalo production in Asia. J Anim and Plant Sci. 22: 250-256.##
Pipalia, D.L., Joshi, C.G., Rank, D.N., Brahmshktri, B.P., Solanki, J.V. (2004) PCR-SSCP typing of MHC in cattle and buffaloes. Ind J Anim Sci. 74: 637-639.##
Rahimnahal, S., Fayazi, J., Mirzadeh, K., Nassiry, M.T., Roshanfekr, H. (2010a) Detection of allelic polymorphism in a gene of the major histocompatibility complex of Iranian buffalo. J Anim Vet Adv. 9: 1902-1904.##
Rahimnahal, S., Fayazi, J., Mirzadeh, K., Nassiry, M.T., Roshanfekr, H. (2010b) Determining BoLA-DRB3 exon 2 polymorphism in buffalo population in Khouzestan province by PCR-RFLP method. Gen Eng Biosafety J. 2: 121-128.##
Rupp, R., Hernandez, A., Mallard, B.A. (2007) Association of  bovine leukocyte antigen (BoLA)DRB3.2 with immune responce, mastitis and production and type traits. Dairy Sci. 90: 1029-1039.##
Sena, L., Schneider, M.P.C., Brenig, B., Honeycutt, R.L., Womack, J.E., Skow L.C. (2003) Polymorphisms in MHC-DRA and -DRB alleles of water buffalo (Bubalus bubalis) reveal diferent features from cattle DR alleles. Anim Genet. 34: 1-10.##
Sheikhmohammadi, R., Hashemi, A., Mardani, K. (2010) Analysis of polymorphism of MHC class II BuLA DRB3 exon 2 gene in North West Iranian populations of the Water buffalo (Bubalusbubalis) through PCR-SSCP. Int J Vet Res. 4: 265-268.##
Takeshima, S.N., Ikegami, M., Morita, M., Nakaim, Y., Aida, Y. (2001) Identification of new cattle BoLA-DRB3 alleles by sequence-based typing. J Immunogenet. 53: 74-81.##
Tizard, I.R. (2009) Acquired immunity: Antigen-presenting receptors. In: Veterinary Immunology., Saunders (7th ed.) Philadelphia. USA. p. 67-76.##
Van Eijk, M.J., Stewart-Haynes, J.A., Lewin, H.A. (1992) Extensive polymorphism of the BoLA-DRB3 gene distinguished by PCR-RFLP. Anim Genet. 23: 483-496.##
Wenink, P.W., Groen, A.F., Roelke-Parker, M.E., Prins, H.H. (1998) African buffalo maintain high genetic diversity in the major histocompatibility complex in spite of historically known population bottlenecks. Mol Ecol. 7: 1315-132.##