DETECTION OF BOVINE LEUKOCYTE ADHESION DEFICIENCY, DEFICIENCY OF URIDINE MONOPHOSPHATE SYNTHASE, AND COMPLEX VERTEBRAL MALFORMATION IN HOLSTEIN CATTLE

Abstract

This research aimed to evaluate the prevalence of the most common lethal diseases in the Lithuanian Holstein cattle population. Two hundred non-related (based on the documentation of origin) cattle (cows and heifers) were included in the study. DNA extraction from blood leukocytes was performed using the chloroform salt method. The cattle were tested for three inherited bovine disorders: bovine leukocyte adhesion deficiency (BLAD), deficiency of uridine monophosphate synthase (DUMPS), complex vertebral malformation (CVM). The PCR-RFLP test method was used to determine the polymorphism of the CD18 gene, which is responsible for BLAD inherited disorder development. A recessive allele with point mutation A→G (383), causing BLAD, was found in the Lithuanian cattle population with 0.0025 frequency. CVM disease is determined by the missense mutation, which has been found in the SLC35A3 gene. The study was performed using a sequencing method. A recessive allele with point mutation G→T (538), causing CVM, was found in the Lithuanian cattle population with 0.005 frequency. The PCR-RFLP test method was used to determine the polymorphism of the UMPS gene, responsible for DUMPS inherited disorder development. A recessive allele with point mutation C→T (1213), causing DUMPS, was not found in the Lithuanian Holstein cattle population. Because intensive selection programmes were performed over the previous decade, the number of heritable lethal diseases carriers has significantly decreased. Key words: cattle; bovine leukocyte adhesion deficiency; deficiency of uridine monophosphate synthase; complex vertebral malformation; BLAD; DUMPS; CVM UGOTAVLJANJE POMANJKLJIVE ADHEZIJE GOVEJIH LEVKOCITOV, POMANJKANJA ENCIMA URIDIN MONOPOSFAT SINTETAZE TER KOMPLEKSA MALFORMACIJE VRETENC PRI HOLŠTAJNSKEM GOVEDU Povzetek: Namen raziskave je bil oceniti razširjenost najpogostejših smrtnih bolezni v populaciji litvanskega holštajnskega goveda. V študijo je bilo vključenih dvesto nesorodnih (na podlagi dokumentacije o poreklu) krav in telic. Izolacijo DNK iz krvnih levkocitov smo izvedli z metodo izolacije s soljo in kloroformom. V vzorcih DNK krav in telic smo preverili prisotnost mutacij v treh genih, ki povzročajo naslednje bolezni: motnjo prilepljanja govejih levkocitov (BLAD), pomanjkanje encima uridin monofosfat sintaze (DUMPS) ter kompleksno malformacije vretenc(CVM). Metoda PCR-RFLP je bila uporabljena za določanje polimorfizma gena CD18, ki je odgovoren za razvoj dedne bolezni BLAD. Recesivni alel s točkovno mutacijo A → G (383), ki povzroča bolezen BLAD, je bil ugotovljen v populaciji litvanskega goveda s frekvenco pojavljanja 0,0025. Bolezen CVM je povzročena z drugačno-pomensko mutacijo v genu SLC35A3. Prisotnost mutacije v tem genu smo izvedli z metodo sekvenciranja. Recesivni alel s točkovno mutacijo G → T (538), ki povzroča CVM, je bil ugotovljen v populaciji litvanskega goveda s frekvenco pojavljanja 0,005. Testna metoda PCR-RFLP je bila uporabljena za določanje polimorfizma gena UMPS, ki je odgovoren za razvoj dedne bolezni DUMPS. Recesivni alel s točkovno mutacijo C→ T (1213), ki povzroča DUMPS, ni bil najden v litvanski populaciji holštajnskega goveda. Ker so se v preteklem desetletju izvajali intenzivni selekcijski programi, se je število prenašalcev dednih smrtnih bolezni znatno zmanjšalo. Ključne besede: govedo; moteno prilepljanje govejih levkocitov; pomanjkanje encima uridin monofosfat sintaze; kompleksna malformacije vretenc; BLAD; DUMPS; CVM