Identification of Holstein heifer’s carriers BLAD and Citrullinemia in the eastern region of Uruguay by PCR-RFLP and sequencing

Abstract

Deficiency in bovine leukocyte adhesion (BLAD) and arginosuccinate synthase enzyme (Citrullinemia) are autosomal recessive diseases that have been described worldwide in the Holstein breed. The objective of this study was to optimize and implement a methodology of genotyping for the identification of animals carrying the allele causing these diseases in a population cohort of calves Holstein of dairy farm in Uruguay basin. DNA samples from 190 Holstein calves were extracted from fresh blood following rules and protocols DNA Bank Unit Biotechnology INIA Las Brujas. Genotyping was performed by PCR-RFLP analysis with restriction enzymes TaqI for BLAD and Eco47I (AvaII) for Citrullinemia. Results of RFLP genotyping were confirmed by sequencing of the amplified products of both diseases. The presence of the BLAD mutant allele was detected in only one calf while no Citrullinemia carriers were found in the analyzed population. This study represented the first survey of the prevalence of BLAD and Citrullinemia diseases of dairy cattle in the eastern region of Uruguay.