Association of DNA polymorphisms of the growth hormone and prolactin genes with milk productivity in Yaroslavl and black-and-white cattle

Polymorphisms of the prolactin (bPRL) and growth hormone (bGH) genes were studied comparatively in the Russian and German Black-and-White and Yaroslavl cattle breeds. Two polymorphisms were studied for each gene. In the case of the bPRL gene, the polymorphism of the 5'-untranslated region was examined by microsatellite analysis and the RsaI polymorphism of exon 3, by RFLP analysis. In the case of the bGH gene, the MspI polymorphism of intron III and the AluI polymorphism of exon 5 were assessed by RFLP analysis. Differences in allele and genotype frequencies were observed both between and within breeds. The heterozygosity at the RsaI marker was low (9.4%) in the Russian Black-and-White breed; that at the microsatellite of the bPRL gene was low (3.2-24%) in all breeds examined. Homozygotes BB at the bPRL gene, which had not been reported earlier for European cattle breeds, were detected in the German Black-and-White and Yaroslavl breeds (at frequencies 0.16 and 0.13, respectively). The frequency of allele MspI(-) of the bGH gene in the Yaroslavl breed was extremely low (0.02), comparable only with that of the Holstein cattle (0.02). The heterozygosity at the AluI polymorphism was higher than at the MspI polymorphism of the bGH gene and reached 55% in the Yaroslavl breed. Genotype BB of the RsaI polymorphism of the bPRL gene tended to show a negative association with the fat content in milk. The genotypes of the AluI polymorphism of the bGH gene were associated with the fat content in milk in the Yaroslavl (F = 4.56, P = 0.013) and German Black-and-White (F = 4.1, P = 0.014) breeds: the highest fat content in milk was observed in the subsample of cows with heterozygous genotype VL.     

Association of DNA polymorphisms of the growth hormone and prolactin genes with milk productivity in Yaroslavl and Black-and-White cattle

Polymorphisms of the prolactin (bPRL) and growth hormone (bGH) genes were studied comparatively in the Russian and German Black-and-White and Yaroslavl cattle breeds. Two polymorphisms were studied for each gene. In the case of the bPRL gene, the polymorphism of the 5-untranslated region was examined by microsatellite analysis and the RsaI polymorphism of exon 3, by RFLP analysis. In the case of the bGH gene, the MspI polymorphism of intron III and the AluI polymorphism of exon 5 were assessed by RFLP analysis. Differences in allele and genotype frequencies were observed both between and within breeds. The heterozygosity at the RsaI marker was low (9.4%) in the Russian Black-and-White breed; that at the microsatellite of the bPRL gene was low (3.2–24%) in all breeds examined. Homozygotes BB at the bPRL gene, which had not been reported earlier for European cattle breeds, were detected in the German Black-and-White and Yaroslavl breeds (at frequencies 0.16 and 0.13, respectively). The frequency of allele MspI(–) of the bGH gene in the Yaroslavl breed was extremely low (0.02), comparable only with that of the Holstein cattle (0.02). The heterozygosity at the AluI polymorphism was higher than at the MspI polymorphism of the bGH gene and reached 55% in the Yaroslavl breed. Genotype BB of the RsaI polymorphism of the bPRL gene tended to show a negative association with the fat content in milk. The genotypes of the AluI polymorphism of the bGH gene were associated with the fat content in milk in the Yaroslavl (F=4.56, P=0.013) and German Black-and-White (F=4.1, P=0.041) breeds: the highest fat content in milk was observed in the subsample of cows with heterozygous genotype VL.Translated from Genetika, Vol. 41, No. 2, 2005, pp. 229–236.Original Russian Text Copyright © 2005 by Khatami, Lazebny, Maksimenko, Sulimova.     

Leukocyte acid phosphatase and metabolic efficiency of phagocytes in the first lactation trimester of cows from a leukaemic herd

In Black-and-White cattle, polymorphism of acid phosphatase (AcP) of blood leukocytes is determined by a pair of autosomal alleles. The aim of the study was to determine the relationship between AcP polymorphism and the metabolic efficiency of phagocytes in the first months after calving of cows naturally infected with the bovine leukaemia virus. The studied population consisted of 91 Black-and-White cows aged 3-6 years, from one herd. Enzootic bovine leukaemia (EBL) was diagnosed with the immuno-enzymatic ELISA method and a PCR molecular test. Additionally, agarose gel electrophoresis and the cytochemical method were used to determine the AcP polymorphism and activity in leukocytes. The metabolic activity of phagocytes was determined by the nitroblue tetrazolium (NBT) reduction test. Significant differences in metabolic efficiency of granulocytes were observed between cows representing different AcP phenotypes. No significant differences in levels of the analysed indices were observed between the EBL-positive and EBL-negative cows and between the three subsequent months after calving.     

Study of the association between alleles of the growth hormone receptor and prolactin receptor genes of bulls and the milk productivity of their daughters

A substitution of thyrosine for phenylalanine (F297Y) in the transmembrane domain of the growth hormone receptor (GHR) was tested for significance for breeding evaluation of bulls of the holstenized Black-and-White breed. The breeding value was estimated by the method of daughter yield deviation to contemporaries with modification. The frequency of genotype FF in the bulls examined was 0.37, lower than in Holstein bulls (0.67). The F297Y substitution exerted the greatest effect on the milk fat content (1.5??) and milk yield (0.8 ??) and a lower effect on the milk fat yield (0.6 ??), milk protein yield (0.5 ??), and milk protein content (0.6 ??). The GHR4.2 single nucleotide substitution (SNP) in the promoter of the GHR gene did not affect the milk production traits. A substitution of asparagine for serine (S18N) in the transmembrane domain of the prolactin receptor (PRLR) was also examined, but it did not significantly affect the milk production traits. The results are discussed in the context of the hypothesis that multiplicity of causal mutations of a particular gene is common and should be taken into account in the genetics of quantitative traits.     

Restriction fragment length polymorphism of bovine lysozyme genes

Genetic polymorphism of bovine lysozyme (LYZ) genes was investigated by analysing restriction fragment length polymorphism (RFLP). The analysis revealed three RFLP loci designated LYZ1, LYZ2 and LYZ2. Each system included two or three allelic variants. Evidence for close genetic linkage of the three loci was found. There was also a significant linkage disequilibrium among the three loci in a sample of about 200 breeding bulls from one breed. No statistically significant association was found between LYZ RFLPs and breeding values of bulls for disease or milk production traits.     

The genetic polymorphism of the blood proteins in RID-positive reacting cows]

Cattle herd of Black-and-White, Red and Simmental breeds having positive or negative RID-reaction to leucosis was studied as to the polymorphism of serum blood proteins in three loci: Tf, Am and Cp. One system of polymorphic proteins has been determined as having a higher concentration of homozygotes (Tf) and another one as having a higher concentration of heterozygotes (Am) within one and the same herd among animals with the positive RID-reaction.     

Two-dimensional electrophoresis of horse serum proteins: genetic polymorphism of ceruloplasmin and two other serum proteins

Two-dimensional agarose gel (pH 8.6)-horizontal polyacrylamide gel (pH 9.0) electrophoresis of horse serum proteins revealed genetic polymorphism of ceruloplasmin (Cp) and two unidentified serum proteins tentatively designated serum protein 1 (SP1) and serum protein 2 (SP2). Family data were consistent with the hypothesis that the observed Cp and SP1 phenotypes were each controlled by two co-dominant, autosornal alleles. The three common SP2 phenotypes were shown to be controlled by two codominant, autosomal alleles. Population data and limited family data indicated the occurrence of two additional SP2 alleles. Altogether more than 600 horses representing 13 different breeds were typed for Cp, SP1 and SP2, and allele frequency estimates were calculated. SP2 was highly polymorphic in all breeds studied whereas SP1 and Cp showed quite low degrees of polymorphism. SP1 polymorphism was observed in seven breeds while Cp polymorphism was observed only in the Icelandic toelter horse breed.     

Gelsolin expression in sheep milk somatic cells during lactation

The identification of genes involved in phenotypes related to milk quality is important for both economic and health aspects in livestock production. The aim of this study was to assess the level of gelsolin gene expression in two breeds of dairy sheep – Sarda and Gentile – with pronounced differences in quantitative and qualitative milk traits. Gelsolin, a type of actin-modulating proteins is involved in the processes of actin remodeling during cell growth and apoptosis; therefore a role of this protein in mammary changes during lactation was here hypothesized. Individual milk samples were collected three times during lactation from 26 ewes of the two breeds. The differential gene expression of gelsolin in the two breeds and the three lactation times was estimated by quantitative PCR on RNA extracted from milk somatic cells. Correlations of gelsolin gene expression with milk yield and quality and days of lactation were also estimated. The results showed that gelsolin gene expression was significantly higher in the Sarda compared to the Gentile at each lactation stage, in agreement with the longer lactation duration and the higher daily milk yield of the first breed. Significant correlations of gelsolin gene expression were found with milk fat content in Sarda breed (−0.46, P<0.05). Gelsolin expression analysis confirmed the link between gelsolin gene function and milk fat content of sheep.     

Nucleotide sequence polymorphism within exon 4 of the bovine prolactin gene and its associations with milk performance traits

Prolactin plays an important regulatory function in mammary gland development, milk secretion, and expression of milk protein genes. Hence the PRL gene is a potential quantitative trait locus and genetic marker of production traits in dairy cattle. We analysed the sequence of the PRL gene to investigate whether mutations in this sequence might be responsible for quantitative variations in milk yield and composition. Using SSCP and direct sequencing, we detected six single-nucleotide polymorphisms within a 294-bp prolactin gene fragment involving exon 4. All detected mutations were silent with respect to the amino acid sequence of the protein. PCR-RFLP genotyping of SNP 8398 R (RsaI) was used to assess allele frequencies in 186 Black-and-White cows (0.113 and 0.887 for A and G, respectively) and in 138 Jersey cows (0.706 and 0.294 for A and G, respectively). Black-and-White cows with genotype AG showed the highest milk yield, while cows with genotype GG showed the highest fat content.     

Polymorphism of alkaline ribonuclease in the leucocytes of Black-and-White cattle

Studies were carried out on the polymorphism of alkaline ribonuclease in Black-and-White cattle in the north-eastern region of Poland. Leucocytes isolated from peripheral blood were used. Three phenotypes were defined, consisting of the combination of two alleles. The gene frequency was: q⁴= 0,7082, q^B= 0,2918. In calves the frequency of allele RNA-ase^B and number of RNA-ase^A/ RNA -ase^B heterozygotes were higher than in adult animals.     

Frequencies of alleles and genotypes of the PRNP gene in Polish Red, Czech Pied and Czech Black-and-White cattle

Frequencies of alleles and genotypes of the PRNP gene in 65 Polish Red cows kept under a conservation programme and 52 randomly chosen cows of two Czech breeds: Czech Pied (42) and Black-and-White (10) were studied. All cows were at the age of 4 years or older. It was found that the frequency of allele 5 (5 copies of the octapeptide repeat) ranged from 0.11 (Polish Red) to 0.00 (Czech Black-and-White) whereas the frequency of allele 6 (6 copies of the octapeptide repeat) ranged from 1.00 (Czech Black and White) to 0.89 (Polish Red). The highest frequency was found for the homozygous genotype 6/6 (1.00 in Czech Black-and-White) and the lowest frequency was detected for the heterozygous genotype 6/5 (0.143). In the studied cows the genotype 5/5 was not found. The higher frequency of allele 5 in the native breeds and its lower frequency or lack of this allele in the populations intensively selected for high milk production may suggest that alleles of the PRNP gene may be associated with milk production.     

犬MC1R基因T105A基因座多态性及 与毛色性状相关性的研究The

为了检测犬MC1R基因T105A基因座的多态性,并分析该多态性与犬毛色表型的相关性,抽取111只外科手术学实验用杂种犬血液并提取DNA,记录毛色表型。采用PCR-RFLP技术,对MC1R基因T105A基因座进行基因多态性分析,并对该基因座DNA进行克隆测序;用二元变量相关分析的统计学方法分析基因座多态性与毛色性状之间的相关性。经PCR-RFLP分析结果表明,T105A基因座序列具有多态性,表现为A、B二个等位基因和AA、AB及BB 3种基因型。A、B等位基因频率分别为72.97%和27.03%,基因杂合度（H）为0.39。基因型AA频率为55.86%,BB为9.91%,AB为34.23%。对T105A多态性片段DNA克隆测序后发现,MC1R基因在编码第105位氨基酸的密码子第一个碱基存在由G到A的单碱基突变,该突变导致第105位氨基酸发生由丙氨酸向苏氨酸的改变。统计分析结果表明MC1R基因T105A基因座的多态性与毛色性状不存在显著的相关性,这可能是由于外科手术学实验用犬是杂种犬,其遗传背景不同所致,尚须在纯种犬群体中进一步研究MC1R基因对毛色的影响。 Abstract: In order to detect the polymorphism of T105A in MC1R gene in dogs and to analyze the relationship between the genetic polymorphisms and phenotypes of dog coat color, the blood samples of 111 cross-breed dogs were taken and their genomic DNAs were extracted. The phenotypes of dog coat color were recorded. The T105A locus of MC1R gene in the canine was detected through the technology of PCR-RFLP. Furthermore, the polymorphic fragments at T105A were sequenced. The relationships between the polymorphism of T105A and coat color trait were analyzed by the statistical methods of bivarate correlation analysis. By the method of PCR-RFLP, the T105A polymorphism was found with two alleles A and B and three genotypes AA, AB and BB. The frequencies of two alleles were 72.97% and 27.03%, respectively. The heterozygosity of T105A locus was 0.39. The frequencies of three genotypes were 55.86%, 34.23% and 9.91%, respectively. According to the results of sequencing, one base change from G to A at the position 105 was found at T105A locus and it altered amino acid at the position 105 from alanine to threonine. According to the statistical analysis, no significant association between the polymorphism of MC1R gene and the coat color was found and the result may be due to the differences of genetic background. Further research on MC1R gene should be done in pure breed dogs.     

Genetic diversity of protein markers in sheep population from Ukraine

Based on polymorphism of genes for antigen factors of six blood-group systems and four blood protein loci, genetic structure and the main variation parameters were studied in three sheep breeds and three sheep breed types constituting the basis of purebred sheep resources in Ukraine. Specific features of the distribution of genotypes and alleles of polymorphic loci were determined in each of the studied sheep groups depending on their origin and production type. The molecular-genetic markers used in the analysis of the genetic relationships between the sheep breeds and breed types were shown to objectively reflect their breeding history and evolution. Integrally, each of the studied gene pools had a specific profile of gene frequencies reflecting breeding specificity, breed history, and genetic differentiation of breeds.     

Genetic Diversity of Protein Markers in Sheep Populations from Ukraine

Based on polymorphism of genes for antigen factors of six blood-group systems and four blood protein loci, genetic structure and the main variation parameters were studied in three sheep breeds and three sheep breed types constituting the basis of purebred sheep resources in Ukraine. Specific features of the distribution of genotypes and alleles of polymorphic loci were determined in each of the studied sheep groups depending on their origin and production type. The molecular–genetic markers used in the analysis of the genetic relationships between the sheep breeds and breed types were shown to objectively reflect their breeding history and evolution. Integrally, each of the studied gene pools had a specific profile of gene frequencies reflecting breeding specificity, breed history, and genetic differentiation of breeds.     

Associations between blood groups, blood protein polymorphisms and breeding values for production traits in Swedish Red and White Dairy bulls

The relationships of nine blood group systems and two blood protein polymorphisms with breeding values for several production traits were examined in dairy cattle of the Swedish Red and White (SRB) breed. The material consisted of 2212 bulls; the bulls were performance tested for growth rate and their breeding values for milk yield, fat and protein content in milk were estimated from progeny tests. The direct effect of marker alleles or marker phenotypes was analysed in a multiple regression model. Several significant associations were found; many supported earlier findings. However, the contribution of the markers to the total variation of the breeding values was very small. Linkage between marker loci and production loci was studied in offspring from heterozygous sires by estimating the interactions between sire and marker alle using a model eliminating the direct effects of sire and marker alle. There were strong indications of linkage between some marker loci (e.g. B, J and Am-1 loci) and loci with large effects on production traits.     

Suitability of cross-bred cows for organic farms based on cross-breeding effects on production and functional traits

Data from 113 Dutch organic farms were analysed to determine the effect of cross-breeding on production and functional traits. In total, data on 33 788 lactations between January 2003 and February 2009 from 15 015 cows were available. Holstein–Friesian pure-bred cows produced most kg of milk in 305 days, but with the lowest percentages of fat and protein of all pure-bred cows in the data set. Cross-breeding Holstein dairy cows with other breeds (Brown Swiss, Dutch Friesian, Groningen White Headed, Jersey, Meuse Rhine Yssel, Montbéliarde or Fleckvieh) decreased milk production, but improved fertility and udder health in most cross-bred animals. In most breeds, heterosis had a significant effect (P < 0.05) on milk (kg in 305 days), fat and protein-corrected milk production (kg in 305 days) and calving interval (CI) in the favourable direction (i.e. more milk, shorter CI), but unfavourably for somatic cell count (higher cell count). Recombination was unfavourable for the milk production traits, but favourable for the functional traits (fertility and udder health). Farm characteristics, like soil type or housing system, affected the regression coefficients on breed components significantly. The effect of the Holstein breed on milk yield was twice as large in cubicle housing as in other housing systems. Jerseys had a negative effect on fertility only on farms on sandy soils. Hence, breed effects differ across farming systems in the organic farming and farmers can use such information to dovetail their farming system with the type of cow they use.     

Analysis of genetic variability at codon 42 within caprine <Emphasis Type="Italic">prion protein</Emphasis> gene in relation to production traits in Chinese domestic breeds

In this study, genetic variability at codon 42 within prion protein (PRNP) gene and its associations with production traits were investigated in 2002 goats from four Chinese domestic breeds. The frequencies of allele “A” ranged from 0.353 to 0.562 in analyzed goat breeds with Hardy–Weinberg equilibrium (P > 0.05) except Xinong Sannen (XNSN) dairy breed. The establishment of relationships between different genotypes and growth traits was performed in Inner Mongolia white Cashmere (IMWC) breed and revealed an association of the polymorphism with body weight at 7-year-old goats (P = 0.033). The individuals with genotype GG showed heavier body weight than those with genotype AA. Moreover, association analysis detected two significant associations between different genotypes and cashmere yield and fiber length in IMWC breed (P = 0.009, P = 0.048, respectively). In addition, three significant associations of different genotypes with density of milk (a.m. and p.m.), solids-not-fat of milk (P = 0.013, P = 0.009 and P = 0.002), respectively, were found in XNSN breed. Genotype GG had better milk quality than others. These findings suggested that the polymorphism of codon 42 within PRNP was a useful DNA marker for eliminating or selecting excellent individuals in relation to production traits in marker-assist selection breeding of goat.     

Variation in blood serum proteins and association with somatic cell count in dairy cattle from multi-breed herds

Blood serum proteins are significant indicators of animal health. Nevertheless, several factors should be considered to appropriately interpret their concentrations in blood. Therefore, the objectives of this study were (1) to assess the effect of herd productivity, breed, age and stage of lactation on serum proteins and (2) to investigate association between serum proteins and somatic cell count (SCC) in dairy cattle. Milk and blood samples were collected from 1508 cows of six different breeds (Holstein Friesian, Brown Swiss, Jersey, Simmental, Rendena and Alpine Grey) that were housed in 41 multi-breed herds. Milk samples were analyzed for composition and SCC, while blood samples were analyzed for serum proteins (i.e. total protein, albumin, globulin and albumin-to-globulin ratio (A : G)). Herds were classified as low or high production, according to the cow’s average daily milk energy yield adjusted for breed, days in milk (DIM) and parity. Data were analyzed using a linear mixed model that included the fixed effects of DIM, parity, SCS, breed, herd productivity and the random effect of the Herd-test date within productivity level. Cows in high producing herds (characterized also by greater use of concentrates in the diet) had greater serum albumin concentrations. Breed differences were reported for all traits, highlighting a possible genetic mechanism. The specialized breed Jersey and the two dual-purpose local breeds (Alpine Grey and Rendena) had the lowest globulin concentration and greatest A : G. Changes in serum proteins were observed through lactation. Total protein reached the highest concentration during the 4th month of lactation. Blood albumin increased with DIM following a quadratic pattern, while globulin decreased linearly. As a consequence, A : G increased linearly during lactation. Older cows had greater total protein and globulin concentrations, while albumin concentration seemed to be not particularly affected by age. A linear relationship between serum proteins and SCS was observed. High milk SCS was associated with greater total protein and globulin concentrations in blood. The rise in globulin concentration, together with a decrease in albumin concentrations, resulted in a decline in A : G as SCS of milk increased. In conclusion, such non-genetic factors must be considered to appropriately interpret serum proteins as potential animal welfare indicator and their evaluation represents an important first-step for future analysis based on the integration of metabolomics, genetic and genomic information for improving the robustness of dairy cows.