A. Lundén

Linkage relationships in the bovine MHC region. High recombination frequency between class II subregions.

Class II genes of the bovine major histocompatibility complex (MHC) have been investigated by Southern blot analysis using human DNA probes. Previous studies revealed the presence of bovine DOβ, DQα, DQβ, DRαand DRβgenes, and restriction fragment length polymorphisms for each of these genes were documented. In the present study, the presence of three additional class II genes, designated DZα, DYα, and DYβ, are reported. DZαwas assumed to correspond to the human DZαgene while the other two were designated DY because their relationship to human class II genes could not be firmly established. The linkage relationships among bovine class II genes and two additional loci, TCP1B and C4, were investigated by family segregation analysis and analysis of linkage disequilibrium. The results clearly indicated that all these loci belong to the same linkage group. This linkage group is divided into two subregions separated by a fairly high recombination frequency. One region includes the C4, DQα, DQβ, DRαand DRβloci and the other one is composed of the DOβDYα, DYβ, and TCPIB loci. No recombinant was observed within any of these subregions and there was a strong or fairly strong linkage disequilibrium between loci within groups. In contrast, as many as five recombinants among three different families were detected in the interval between these subregions giving a recombination frequency estimate of 0.17 ± 0.07. The fairly high recombination frequency observed between class 11 genes in cattle is strikingly different from the corresponding recombination estimates in man and mouse. The finding implies either a much larger molecular distance between some of the bovine class II genes or alternatively the presence of a recombinational “hot spot” in the bovine class II region.

Effect of β-casein, κ-casein and β-lactoglobulin genotypes on concentration of milk protein variants

Individual mid-lactation milk samples were collected from 116 cows of the Swedish Red and Swedish Holstein breeds with known genotypes of beta-casein, kappa-casein and beta-lactoglobulin. Detailed milk protein composition and allele-specific expression of beta-casein, kappa-casein and beta-lactoglobulin proteins in milk were analysed using reversed-phase high-performance liquid chromatography. Aggregate beta-/kappa-casein genotype was significantly associated with the kappa-casein concentration in milk. The lowest kappa-casein concentration was found in milk of cows with genotypes including kappa-casein E (A(1)A(2)/AE, A(1)A(1)/AE) and also A(2)A(2)/AA milk, whereas highest levels were associated with genotypes including kappa-casein B. Casein number was positively and concentration of beta-lactoglobulin negatively associated with the beta-lactoglobulin BB genotype. In heterozygote cows, beta-casein A(1) and beta-lactoglobulin A proteins were found at higher concentrations in milk compared with the protein variant encoded by the alternative allele at these loci, whereas kappa-casein A and B variants were found at similar concentrations in heterozygote AB cows.

Association of polymorphisms at DGAT1, leptin, SCD1, CAPN1 and CAST genes with color, marbling and water holding capacity in meat from beef cattle populations in Sweden

The objective of this study was to investigate the associations of single nucleotide polymorphisms (SNPs) at five candidate genes with meat pH, color, marbling and water holding capacity (WHC) in young bulls from five beef breeds (n=243) in Sweden. The UASMS2 polymorphism at the leptin gene and the SNPs at the Stearoyl-CoA desaturase 1 gene (SCD1.878) and μ-calpain gene (CAPN1:c.947) were associated with variation in meat color traits after 6days of exposure to air. The K232A polymorphism at the diacylglycerol O-acyltransferase 1 (DGAT1) gene and the CAPN1:c.947 SNP were associated with level of beef marbling. There was no association between the SNP at the calpastatin gene (CAST:c.155) and meat quality traits, nor was there any association of the tested SNPs with WHC traits and pH value.

Casein retention in curd and loss of casein into whey at chymosin-induced coagulation of milk

Impact of milk protein composition on casein (CN) retention in curd during the milk coagulation process was studied using a model cheese making system. Individual milk samples from 110 cows in mid lactation of the Swedish Red and Swedish Holstein breeds with known genotypes of beta-casein, kappa-casein and beta-lactoglobulin were defatted, coagulated with chymosin, subjected to syneresis and subsequent pressing simulated by centrifugation. The results indicated that kappa-casein concentration of milk plays an important role in the curd formation process and initial syneresis (whey after cutting), whereas an increased CN ratio was associated with less casein in whey after simulated pressing. Increased kappa-casein concentration of milk also characterized the milk samples with no measurable loss of casein in whey, compared with milk samples with casein lost in whey, both after cutting and after simulated pressing. Concentrations of alphas1-casein, beta-casein, and total casein in milk were positively associated with fresh curd yield, which showed a strong correlation with amount of casein retained in curd. No effect of protein genotype on fresh curd yield or casein in whey was found. The beta-lactoglobulin BB genotype was associated with increased casein retention in curd, most likely due to the association of this genotype with CN ratio.

Composition of poorly and non-coagulating bovine milk and effect of calcium addition

Ninety-nine individual milk samples from 37 cows in lactation week 10-35, selected for producing well or poorly/non-coagulating milk, were compared regarding protein composition, total calcium content, casein micelle size, pH, and coagulating properties after addition of 0·05% CaCl₂. The results showed that a low κ-casein concentration in milk was a risk factor for non-coagulation. CaCl₂ addition improved coagulating properties (coagulation time, curd firmness) of nearly all samples and eliminated differences between poorly/non-coagulating and well-coagulating milk, particularly regarding curd firmness. A second, independent data set with 18 non-coagulating or well-coagulating milk samples were analysed for protein composition, where indications of a similar association with κ-casein was observed.

Relative impact of α-tocopherol, copper and fatty acid composition on the occurrence of oxidized milk flavour

The oxidative process in milk fat, resulting in spontaneous oxidized off-flavour (SOF), is commonly assumed to depend on contents of pro- and antioxidants in milk and availability of fatty acids acting as their substrate. An important antioxidant in milk is alpha-tocopherol whereas the most potent prooxidant is the metal ion copper. The separate effects of alpha-tocopherol, copper, and milk fatty acid profile, and their combined effect on SOF development were examined in milk from 44 multiparous cows fed different roughage types and different amounts of dietary, unsaturated fat. A clear association between concentrations of copper and poly-unsaturated fatty acids in milk and the risk for developing SOF was found. Heritability estimates suggest that occurrence of SOF is partly under genetic control which indicates that milk quality may be compromised if breeding bulls are selected that carry genotypes predisposing for milk prone to develop SOF.

Genetic analyses of pathogen-specific mastitis.

The aims of this study were to investigate the presence of genetic variation for susceptibility to pathogen-specific mastitis and to examine whether haplotypes of an identified quantitative trait locus with effect on unspecific mastitis resistance had different effects on specific mastitis pathogens. Bacteriological data on mastitis pathogens were obtained from the diagnostic laboratory at the Swedish National Veterinary Institute. The data were mainly from subclinical cases of mastitis but also clinical cases were included. Variance components were estimated for incidence of the six most frequent pathogens using Markov Chain Monte Carlo methodology via Gibbs sampling. Genetic variation for susceptibility to pathogen-specific mastitis was higher compared to estimates of general resistance to clinical mastitis in most other studies. However, because of the non-random nature of data collection, comparisons to other studies should be made by caution. The effect of haplotype on the risk of being infected by a given mastitis pathogen, relative to other pathogens, was studied using an allele substitution model. Although there were no significant haplotype substitution effects on the resistance to any of the six mastitis pathogens, there was a significant difference between the effects of two of the haplotypes regarding the risk of acquiring a Streptococcus dysgalactiae infection.

Association of DGAT1 genotype, fatty acid composition, and concentration of copper in milk with spontaneous oxidized flavor

In 136 cows with altogether 969 milk samples, we investigated the effect of the acyl-coenzyme A:diacylglycerol acyltransferase 1 (DGAT1) K232A polymorphism on milk fatty acid (FA) composition and how, in combination with copper concentration in milk, this influences the occurrence of spontaneous oxidized flavor. All milk samples were analyzed for concentrations of copper and individual FA and subjected to sensory analysis by trained judges. We found an effect of DGAT1 genotype on FA composition where mainly the long-chain FA were affected. The 232A allele was associated with larger proportions of the C18:2 cis-9,trans-11 conjugated linoleic acid and lower proportions of C16:0 FA. Milk concentrations of unsaturated FA and copper showed strong and unfavorable associations with spontaneous oxidized flavor (SOF) development. The interaction between FA and copper indicates that SOF will not develop as easily in milk with high copper content unless the substrate is available (i.e., in addition to the previously shown effect of copper in milk, unsaturated FA are required for the process of oxidation to progress). We observed a marked effect of the DGAT1 genotype on SOF development where the A allele was associated with a higher risk of SOF. Moreover, our results suggest that the effects of the FA C18:3 n-3 and of the polyunsaturated index on SOF development are beyond the effect of the DGAT1 genotype. Breed had an effect on FA composition but not on SOF development. Our results imply that copper, FA composition, and DGAT1 genotype are risk factors for SOF and considerations to these factors might be necessary in future breeding decisions.

Associations between candidate SNPs in the calpain 1, calpastatin and leptin genes and meat tenderness among Swedish beef populations

Abstract The contribution of three candidate genes to the variation in meat tenderness was tested in muscle samples from 243 pure-bred, young, beef bulls of Angus, Charolais, Hereford, Limousin and Simmental breeds, raised in Swedish commercial herds. The animals were genotyped for single nucleotide polymorphisms (SNPs) in the calpain 1 (CAPN1), calpastatin (CAST) and leptin genes. The frequent calpain 1 CAPN1:c.947G>C G allele showed an unfavourable association with tenderness. The calpastatin CAST:c.155C>T T allele, which was the most common allele, showed a favourable association with Warner–Bratzler shear force (WBSF) and compression tests. An association was observed between the leptin UASMS2C>T SNP and compression tests.