Investigation of Bovine Leukocyte Adhesion Deficiency (BLAD) and Complex Vertebral Malformation (CVM) in a Population of Iranian Holstein Cows

Document Type: Research Article

Authors

1 Department of Animal Science, Karaj Branch, Islamic Azad University, Karaj, Iran

2 Department of VeterinaryMedicine, Shahre-kord Branch, Islamic Azad University, Shahre-kord, Iran

3 Biotechnilogy Laboratory, Animal Breeding Center, Karaj, Iran

Abstract

In the present research, molecular detection of bovine leukocyte adhesion deficiency (BLAD) and complex vertebral malformation (CVM)in a population of Iranian Holstein cows has been carried outusing milk somatic cells by polymerase chain reaction-restriction fragment length polymorphism(PCR-RFLP). The BLAD and CVM are monogenic and autosomal recessive heredity lethal syndrome in Holstein-Friesian cattle. BLAD characterized by affecting the haematopoietic system via reduced expression of the adhesion molecules on neutrophils. CVM characterized by intra-uterine mortality with disorders such as short neck, curved legs, abnormality of ribs and some certain heart abnormalities. In the first step of our research program,tank milk samples from 50 herds were collected. PCR-RFLP was performed to detect a point mutation of both CVM and BLAD genes. After DNA extraction, PCR was amplified using specific primers for 136 bp DNA (CD18 gene) and233 bp DNA (SLC35A3 gene).TaqIand EcoT22I enzymes were used to identify both BLAD and CVM alleles of both genes by digestion of PCR products.In these herds, we did not find any affected herd with the mutant allele of BLAD comparing with a positive evidence but the mutation of SLC35A3 gene found in 17 different herds. In the next step of our study a herd with 120 cows was randomly selected for individual test using blood samples. We showed two cows out of 120 were identified as carriers of this gene. In this herd, the total number of dominant homozygote (AA), heterozygote (Aa) and recessive homozygote (aa) genotypes for CVM were 118, 2 and 0, respectively and the frequency of A and a alleles were 0.992 and 0.008, respectively. The other affected herds will be tested in the next step of our research program.

Keywords


Adamov N., Mitrov D., Esmerov I. and Dovc P. (2013). Detection of recessive mutations (BLAD and DUMPS) in Holstein-Frisian cattle population in Republic of Macedonia. Mac. Vet. Rev. 37(1), 61-68.
Agerholm J.S., Bendixen C., Andersen O. and Arnbjerg J. (2001). Complex vertebral malformations in Holstein calves. J. Vet. Diagn. Invest. 13, 333-336.
Agerholm J.S., Andersen O., Almskou M.B., Bendixen C., Arnbjerg J., Amand G.P., Nielsen U.S., Panitz F. and Petersen A.H. (2004). Evaluation of the inheritance of the complex vertebral malformation syndrome by breeding studies. Acta Vet. Scand. 45, 133-137.
Betka L., Tatjana K. and Vladimir M. (2008). Detection of recessive mutations (CVM, BLAD and Red factor) in Holstein bulls in Slovenia. J. Cent. Eur. Agric. 9, 101-106.
Boujenane I. and Ouhmama K. (2009). Prevalence of BLAD and CVM in Holstein dairy cattle introduced to Morocco. Egyptian J. Anim. Prod. 46(1), 19-26.
Chu Q., Sun D., Yu Y. Zhang Y. and Zhang Y. (2008). Identification of complex vertebral malformation carriers in Chinese Holstein. J. Vet. Diagn. Invest. 20, 228-230.
Ghanem M.E., Akita M., Suzuki T., Kasuga A. and Nishibori M. (2008). Complex vertebral malformation in Holstein cows in Japan and its inheritance to crossbred F1 generation. Anim. Reprod. Sci. 103, 348-354.
Kepenek E.S. (2007). Polymorphism of prolact in (PRL), diacylglycerol acyltransferase (DGAT-1) and bovine solute carrier family 35 member 3 (SLC35A3) genes in native cattle breeds and its implication for Turkish cattle breeding. MS Thesis. Middle East Technical Univ., Ankara.
Kotikalapudi R., Patel R.K., Sunkara P.S.S. and Roy A. (2013). Detection of silent homozygous polymorphism in exon 4 of SLC35A3gene in a Holstein cattle carrier for complex vertebral malformation. Int. J. Vet. Sci. 2(2), 61-64.
Nagahata H., Noda H., Takahashi K., Kurosawa T. and Sonoda M. (1987). Bovine granulocytopathy syndrome: neutrophil dysfunction in Holstein Friesian calves. J. Vet. Med. 34, 445-451.
Nagahata H., Oota H., Nitanai A., Oikawa S., Higuchi H., Nakade T., Kurosava T., Morita M. and Ogawa H. (2002). Complex vertebral malformation in a stillborn Holstein calf in Japan. J. Vet. Med. Sci. 64(12), 1107-1112.
Patel R.K. (2012). Complex vertebral malformation (CVM) in Holstein. Wayamba J. Anim. Sci. 4, 11-19.
Rezaei A.R., Nassiry M.R., Sadeghi B., Shafagh-Motlagh A., Tahmoorespour M. and Valizadeh R. (2009). Implication of complex vertebral malformation and deficiency of uridinemonophosphate synthase on molecular-based testing in the Iranian Holstein bulls population. African J. Biotechnol. 8(22), 6077-6081.
Thomsen B., Horn P., Panitz F., Bendixen E., Petersen A.H., Holm L.E., Nielsen V.H., Agerholm J.S., Arnbjerg J. and Bendixen C. (2006). A missense mutation in the bovine SLC35A3 gene, encoding a UDP-N-acetyl glucosamine transporter, causes complex vertebral malformation. Gen. Res. 16(1), 97-105.
Vatasescu-Balcan R.A., Manea M.A., Georgescu S.E., Dinischiotu A., Tesio C.D. and Costache M. (2007). Evidence of single point mutation inducing BLAD disease in Romanian Holstein-derived cattle breed. Biotechnol. Anim. Husb. 23, 375-381.