Roumanian Biotechnological Letters Vol. 11, No. 5, 2006, pp. 2881-2884 Copyright © 2006 Bucharest University Printed in Romania. All rights reserved Roumanian Society of Biological Sciences ORIGINAL PAPER 2881 Incidence of BLAD and DUMPS carriers in Romanian cattle breeds Received for publication, July 5, 2006 Accepted, August 1, 2006 VĂTĂŞESCU–BALCAN R.*, GEORGESCU S.E.*, MANEA MARIA ADINA*, REBEDEA MARIANA**, DINISCHIOTU ANCA*, COSTACHE MARIETA* *University of Bucharest, Faculty of Biology, Molecular Biology Center, 91-95 Spl. Independentei, 050095 Bucharest, Romania E-mail: georgescu_se@yahoo.com, costache@bio.bio.unibuc.ro **Institute of Biology and Animal Nutrition, Baloteşti, Bucharest, Romania Abstract Bovine leukocyte adhesion deficiency (BLAD) is a genetic disease that affects the hematopoetic system by greatly reduced expression of the heterodimeric β2 integrin, causing many defects in leukocyte function. It is known that BLAD deficiency is an autosomal recessive defect, caused by a point mutation in the gene encoding subunit CD18 for β2 integrin adhesion molecule, which is lethal in the homozygous form [1]. DUMPS (Deficiency of Uridine Monophosphate Synthase) is a hereditary recessive disorder in Holstein cattle causing early embryo mortality during its implantation in the uterus. The only way to avoid the economic losses is early detection of DUMPS carriers. A population from the Romanian Black Spotted cattle was used to investigate the incidence of these genetic diseases in the main dairy breed in Romania. The aim of our work was to optimize the PCR–RFLP analysis as a diagnostic test to identify BLAD and DUMPS carrier cattle. Keywords: Romanian Black Spotted, BLAD, DUMPS, PCR, RFLP, genotype detection Introduction Bovine Leukocyte Adhesion Deficiency (BLAD) is a lethal autosomal recessive disease in Holstein cattle characterized by a greatly reduced level of expression of the β2 heterodimeric integrin. Integrins are adhesion molecules that mediate the entry and passage of neutrophiles across membranes to destroy invading pathogens [6; 9; 10]. The molecular basis of BLAD is a single point mutation (A-G) at position 383 in the cDNA of the CD18 gene. This mutation results in a substitution of a glycine for an aspartic acid at position 128 in the D128G protein [2; 5; 8; 11; 15]. Viana [16] and Shuster [15] also described the existence of the silent point mutation (C-T) at position 775 in the cDNA without phenotypic manifestation. DNA studies with the use of restriction endonuclease TaqI or HaeIII have detected differences between healthy and affected calves. Gerardi [2] observed in regard to the method used that normal animals have fragments of 100, 200 and 300bp, and BLAD – calves have 200 and 400bp bands. Viana [16] described the length of fragments for normal animals (191, 152bp), for BLAD carriers (343, 191 and 152bp), and for affected animals (343bp). Deficiency of Uridine – 5´-Monophosphate Synthase (DUMPS) is a genetic disorder, which interferes with pyrimidine biosynthesis and is inherited as a single, two-allele, autosomal locus [7; 14]. The enzyme uridine – 5´-monophosphate (UMP) synthase catalyses the conversion of orotic acid to UMP, the precursor of all other pyrimidine nucleotides and a normal constituent in the milk of cows and other ruminants [13; 14].