Grzegorz Grzybowski

Deletion/insertion polymorphism of the prion protein gene (PRNP) in Polish Holstein-Friesian cattle

The aim of the present study was to identify the deletion/insertion polymorphism of the bovine prion protein gene (PRNP) within the promoter sequence (23 bp), intron 1 (12 bp) and 3’ untranslated region (14 bp). DNA was isolated from blood of 234 randomly tested Polish Holstein-Friesian cows and from semen of 47 sires used for artificial insemination (AI) in 2004. No statistically significant differences were found in the frequency of genotypes and alleles between cows and breeding bulls in the 3 analysed polymorphic sites within thePRNP gene. Only 3 haplotypes were identified in sires and 4 haplotypes in cows.

Deletion/insertion polymorphism of the prion protein gene (PRNP) in Polish Red cattle, Polish White-backed cattle and European bison (Bison bonasus L., 1758)

The aim of the present study was to identify deletion/insertion polymorphism of the bovine prion protein (PRNP) gene within the promoter sequence (23 bp indel), intron 1 (12 bp indel) and the 3′ end untranslated region (14 bp indel). The experiment was performed on three groups of animals protected under a genetic resources conservation program: 139 Polish Red (PR) cows, 79 Polish White-backed cows and 50 European bison (Bison bonasus L., 1758). White-backed cattle were characterized by a higher frequency of ins/del heterozygotes and a relatively lower frequency of ins/ins homozygotes within the promoter sequence region (23 bp indel), compared to Polish Red cattle. At the polymorphic locus of intron 1 (12 bp indel) the genetic structure of both cattle populations was similar. Monomorphism, expressed by the occurrence of one genotype variant in each of the analyzed sequence regions, was observed in European bison. Five haplotypes were found in Polish White-backed cows, four haplotypes in Polish Red cows and only one in analyzed group of bison. Differences between the observed and expected number of PRNP haplotypes were recorded in Polish Red cattle.

Effectiveness of a program aimed at the elimination of BLAD-carrier bulls from Polish Holstein-Friesian cattle

The molecular basis of BLAD is the D128G mutation of the gene coding for the CD 18 subunit of beta—2 integrin. This mutation is lethal, since homozygous (BL/BL) animals die before they reach sexual maturity. In the 1990s, BLAD was the most widespread genetic disease in HF cattle worldwide. The aim of the present study was to determine the frequency of BLAD carriers among 4645 young breeding bulls in Poland in 1995–2006. The frequency of carriers of the mutated allele showed a clear decreasing trend. The highest frequency (7.9%) was recorded while implementing the BLAD control program (1995–1997). Regular monitoring has enabled a great reduction of this threat to the tested population. Today only sporadic cases of BL/TL heterozygotes are reported (ca. 0.8% in 2004–2006).

Distribution of mitochondrial haplotypes (cytb) in Polish populations of Emys orbicularis (L., 1758)

The European pond turtle, Emys orbicularis, inhabits a wide distribution area in the western Palaearctic. Polish populations of pond turtle represent the nominotypical subspecies Emys orbicularis orbicularis. The mitochondrial DNA haplotype (cytb gene) variation among 131 turtles from 26 locations in five regions of Poland was investigated. Five haplotypes belonging to three distinct lineages were identified. Two clades (I and II) were represented by two haplotypes each, while the other clade (IV) was represented by one haplotype. Three haplotypes were reported for the first time in E. orbicularis. The eastern part of Poland is inhabited exclusively by turtles bearing haplotype Ia. The remaining four sequence variants were recorded in western Poland where only the IIb haplotype is considered endemic. The distribution of the other haplotypes in western Poland could thus reflect past introductions or accidental releases. The authors regarded the two locations (Drzeczkowo and Karpicko) that were first included in the western Poland populations as autochthonous catchment areas of haplotype Ia.

Population differentiation of the European pond turtle (Emys orbicularis) in Poland inferred by the analysis of mitochondrial and microsatellite DNA: implications for conservation

We investigated the genetic diversity of Polish populations of the European pond turtle (Emys orbicularis) using complete sequences of the mitochondrial cytochrome b gene and allelic variation at thirteen microsatellite loci. We collected data from 146 turtles from 28 locations covering most of the species’ range in Poland. Our results showed a low haplotype diversity and high levels of microsatellite diversity in all populations. We applied two Bayesian approaches using the multilocus data and determined relationships of mtDNA haplotypes by constructing a parsimony network. We observed relatively consistent results of the two Bayesian clustering methods and largely concordant differentiation patterns between mitochondrial and microsatellite markers. All eastern Polish populations were assigned in a highly supported eastern group, while the majority of turtles from western Poland were joined in a distinct western group. The most probable number of population clusters corresponded to the number of native mtDNA lineages. Additionally, we detected a hierarchical structure within the eastern lineage splitting the populations into two subgroups. Polish populations generally constitute genetically homogenous groups, characterized by a low population admixture, with only negligible impact from allochthonous turtles.