Validation and long term performance characteristics of a quantitative enzyme linked immunosorbent assay (ELISA) for human anti-PA IgG.

Accurate, reliable and standardized quantification of anti-protective antigen (PA) IgG antibody levels is essential for comparative analyses of anti-toxin immune responses in anthrax cases, recipients of PA-based anthrax vaccines and for evaluation of anti-PA based immunotherapies. We have previously reported the early performance characteristics and application of a quantitative anti-PA IgG enzyme linked immunosorbent assay. The principal application of this assay was in a Phase 4 human clinical trial of anthrax vaccine adsorbed (AVA, BioThrax), the central component of the CDC Anthrax Vaccine Research Program (AVRP) and in humans following bioterrorism associated Bacillus anthracis infection (Quinn et al., 2002; Quinn et al., 2004; Marano et al., 2008). The objective of the AVRP was to determine the feasibility of reducing the number of priming series and booster doses of the licensed Anthrax Vaccine Adsorbed (AVA) (BioThrax®; Emergent BioSolutions, Lansing, MI) and changing the route of administration from subcutaneous (SC) to intramuscular (IM) (Marano et al., 2008). In this paper we report the validation and long term performance characteristics of the assay during its six year application in the AVRP (2002-2008). The critical features are 1) extensive validation of the assay using two standard reference sera; 2) long term stability and 3) consistency of the data for quantitative analysis of human long term anti-PA IgG responses. The reportable value (RV) of the assay was expressed as anti-PA IgG concentration (µg/ml). Accuracy of the assay was high with a percent error (%ER) range of 1.6-11.4%. Overall intra-operator and intermediate precision were high with Coefficients of Variation (%CVs) of 2.5-15.4% and 6.3-13.2%, respectively. The assay demonstrated excellent dilutional linearity for human sera using log(10) transformed data with the slope=0.95 to 0.99, intercept=0.02 to 0.06 and r(2)=0.980-0.987. The assay was robust, tolerating changes in serum incubation temperatures from 35 to 39°C, serum incubation times from 55 to 65min and changes in key reagents. The long-term assay stability over 6years using consecutive reference sera AVR414 and AVR801 demonstrated sustained high accuracy and precision for the assay, confirming its suitability for long term studies of PA protein-based anthrax vaccines.

Short communication: estimates of genetic variation of milk fatty acids in US Holstein cows.

Interest in changing the milk fatty acid profile is growing. However, little is known about the genetic variability of milk fatty acids in the US Holstein population. Therefore, genetic parameters for milk fatty acids were estimated using a single-trait, mixed, linear animal model on 592 individual milk samples from 233 daughters of 53 sires in a cow herd genetically representative of the US Holstein population. Heritability (h(2)) and repeatability (r) estimates +/- standard errors for yields of individual fatty acids ranged from 0.00 +/- 0.08 (C4:0) to 0.43 +/- 0.13 (C12:0) for heritabilities and from 0.21 +/- 0.05 (C18:1) to 0.43 +/- 0.05 (C12:0) for repeatabilities. Saturated (h(2) = 0.23 +/- 0.12; r = 0.36 +/- 0.05) and de novo synthesized fatty acids (C6:0 to C14:0; h(2) = 0.30 +/- 0.13; r = 0.40 +/- 0.05) had numerically higher estimates than did monounsaturated (h(2) = 0.09 +/- 0.09; r = 0.22 +/- 0.05) and polyunsaturated fatty acids (h(2) = 0.08 +/- 0.09; r = 0.27 +/- 0.05). For relative proportions of individual fatty acids, the greatest heritability and repeatability estimates were obtained for C8:0 (h(2) = 0.18 +/- 0.12; r = 0.36 +/- 0.05), C10:0 (h(2) = 0.22 +/- 0.13; r = 0.46 +/- 0.05), C12:0 (h(2) = 0.18 +/- 0.12; r = 0.46 +/- 0.05), C16:0 (h(2) = 0.09 +/- 0.12; r = 0.48 +/- 0.05), C16:1 (h(2) = 0.49 +/- 0.13; r = 0.49 +/- 0.05), and C18:0 (h(2) = 0.24 +/- 0.11; r = 0.39 +/- 0.05). Our results suggest the existence of genetic variability of milk fatty acids, in particular of medium-and long-chain fatty acids (C8:0 to C18:0), which could be used to improve the nutritional and textural properties of milk fat by selective breeding.

Short communication: Composition of milk protein and milk fatty acids is stable for cows differing in genetic merit for milk production.

Changing the composition of milk protein and of milk fatty acids alters nutritional and physical properties of dairy products and their consumer appeal. Genetic selection for milk yield decreases concentrations of milk protein and of milk fat. Little is known, however, about how the decrease affects composition of milk protein and milk fatty acids. The objective of this study was to quantify changes in composition of milk protein and of milk fatty acids in cows differing in genetic merit for milk production. Three measures of genetic merit for milk production were used for each cow: genetic line, parent average predicted transmitting ability (PTA) for milk, and cow milk PTA. Composition of milk protein and milk fatty acids were compared in 448 milk samples from 178 cows representing 2 divergent lines of Holsteins that were bred for high or average PTA for milk and combined milk protein and fat yield. High-line cows (n = 97) produced more milk that contained less fat and had higher proportions of alphaS1-casein in milk protein than did average-line cows (n = 81). We additionally obtained from 233 cows (178 cows representing the 2 genetic lines and 55 cows with ancestors from both genetic lines) the parent average milk PTA and cow milk PTA and compared composition of milk protein and of milk fatty acids in 592 milk samples. Cows whose parent average milk PTA was above or equal to the median of the 233 cows produced more milk that contained less protein and less fat and that tended to have greater proportions of alphaS1-casein in milk protein than cows whose average milk PTA was below the median. Similarly, cows with above or equal median milk PTA of the 233 cows produced more milk that contained less protein and less fat and had greater proportions of alphaS1-casein in milk protein than did cows with below-median milk PTA. Milk fatty acid composition was not consistently different between groups. Therefore, selection for milk yield decreased concentrations of milk protein and milk fat but had little effect on composition of milk protein and milk fatty acids.

Butter composition and texture from cows with different milk fatty acid compositions fed fish oil or roasted soybeans.

Changing the milk fatty acid composition can improve the nutritional and physical properties of dairy products and their acceptability to consumers. A more healthful milk fatty acid composition can be achieved by altering the cow's diet, for example, by feeding supplemental fish oil (FO) or roasted soybeans (RSB), or by selecting cows with a more unsaturated milk fatty acid composition. We examined whether feeding supplemental FO or RSB to cows that had a more unsaturated milk fatty acid composition acted additively to produce butter with improved fatty acid composition and texture. Using a 3 x 3 Latin square design with 2 replications, we fed diets to multiparous Holstein cows (60 to 200 DIM) chosen for producing either more or less unsaturated milk fatty acid composition (n = 6 for each group) for three 3-wk periods. The control diet contained 3.7% crude fat and the 2 experimental diets contained, on a dry matter basis, 0.8% of additional lipids in the form of 0.9% of FO or 5% of RSB. The milk, collected in the third week of feeding, was used to make butter, which was analyzed for its fatty acid composition and physical properties. Dry matter intake, milk yield, and milk composition were not significantly affected by cow diet or by cow selection. Cows that produced a more unsaturated and healthful milk fat prior to the feeding study, according to a "health-promoting index" [HPI = (sum of % of unsaturated fatty acids)/ (%12:0 + 4 x %14:0 + %16:0)], maintained a higher HPI in their butter during the feeding study than did cows with a low HPI. Milk from cows fed supplemental FO or RSB yielded more unsaturated butters with a higher HPI. This butter also was softer when the cows were fed RSB. Feeding RSB to cows chosen for their high milk HPI yielded the most unsaturated butter with the highest HPI and softest texture. Thus, selecting cows with a more health-promoting milk fatty acid composition and feeding supplemental RSB can be used in combination to produce butter that has a consumer-friendly texture and a healthful fatty acid profile.

Genetic basis and risk factors for infectious and noninfectious diseases in US Holsteins. I. Estimation of genetic parameters for single diseases and general health.

Health data collected from 1996 to 1999 from 177 herds in Minnesota and Wisconsin were analyzed to establish genetic basis for infectious and noninfectious diseases. Three types of health traits were targeted. First, available infectious conditions were used to identify animals that are superior in their general immunity (including innate immunity) for infectious diseases. Generalized immunity may be thought of as a combination of immune responses to a variety of immune system challenges. Second, single infectious and noninfectious diseases were analyzed separately. Third, infectious reproductive diseases as one category of related conditions, and cystic ovary disease as one category of 3 related noninfectious ovary disorders were studied. Data were analyzed using a threshold model that included herd, calving year, season of calving, and parity as cross-classified fixed factors; and sire and cow within sires as random effects. Days at risk and days in milk at the beginning of a record were included by fitting the days as continuous covariates in the model. A heritability value of 0.202 +/- 0.083 was estimated for generalized immunity. Heritability values of 0.141 and 0.161 were estimated for uterine infection and mastitis, respectively. Heritability of single noninfectious disorders ranged from 0.087 to 0.349. The amount of additive genetic variance recovered in the underlying scale of noninfectious disorders tended to zero when combining multiple conditions. The study supports combining infectious diseases into categories of interest but we do not recommend the same approach for noninfectious disorders.

Genetic variation in bovine mononuclear leukocyte responses to dexamethasone.

The objective of this study was to determine whether bovine mononuclear leukocytes exhibit genetic variability prior to and after a glucocorticoid hormone challenge in vivo. Test animals included 60 pedigreed Holstein bulls treated on 3 consecutive days with dexamethasone and 5 untreated control bulls. Eight indicator traits of leukocyte responsiveness to dexamethasone included the percentages of circulating B cells, T cells (CD4, CD8, and workshop cluster 1 molecule expressed by bovine gammadelta T cell), major histocompatibility complex (MHC) I and II expressing cells, and mean expressions of surface MHC I and MHC II on circulating cells. Blood for this work was collected from each test bull 10 times before, during, and after dexamethasone administration, with corresponding samples taken for control bulls. Random regression models with treatment-specific serial correlation were applied to the leukocyte data sets to estimate genetic and nongenetic sources of variation in baseline and recovery aspects of the traits. All traits responded predictably to glucocorticoid challenge. Genetic variation was observed in baseline measurements of all traits, with heritability estimates ranging from 0.21 +/- 0.03 to 0.60 +/- 0.06. Genetic variation in linear recovery from nadir values following dexamethasone administration was significant only for percentage CD4, percentage CD8, and for surface expression of MHC II. The genetic covariance between basal and linear recovery was positive and significant for percentage CD4, percentage CD8, and MHC II expression. The bovine lymphocyte antigen DRB3.2 locus accounted for significant proportions of total variation in percentage MHC II cells and MHC I expression. These results suggest that genetic variability exists for important basal and glucocorticoid-modified phenotypes of bovine mononuclear leukocytes, implying that immunocompetence traits impacted by this stress hormone may be enhanced by genetic selection.

Correlations among body condition scores from various sources, dairy form, and cow health from the United States and Denmark.

The objectives of this study were to estimate genetic correlations among body condition scores (BCS) from various sources, dairy form, and measures of cow health. Body condition score and dairy form evaluated during routine type appraisal was obtained from the Holstein Association USA, Inc. A second set of BCS was obtained from Dairy Records Managements Systems (DRMS) and was recorded by producers that use PCDART dairy management software. Disease observations were obtained from recorded veterinarian treatments in several dairy herds in the United States. Estimated breeding values for diseases in Denmark were also obtained. Genetic correlations among BCS, dairy form, and cow health traits in the United States were generated with sire models. Models included fixed effects for age, DIM, and contemporary group. Random effects included sire, permanent environment, herd-year season for health traits, and error. Predicted transmitting abilities (PTA) for BCS and dairy form were correlated with estimated breeding values for disease in Denmark. The genetic correlation estimate between BCS from DRMS and BCS from the Holstein Association USA, Inc., was 0.85. The genetic correlation estimate between BCS and a composite of all diseases in the United States was -0.79, and PTA for BCS was favorably correlated with an index of resistance to disease other than mastitis in Denmark (0.27). Dairy form was positively correlated with a composite of all diseases in the United States (0.85) and was unfavorably correlated with an index for resistance to disease other than mastitis in Denmark (-0.29). Adjustment for protein yield PTA had a minimal affect on correlations between PTA for BCS or dairy form and disease in Denmark. Selection for higher body condition or lower dairy form with continued selection for yield may slow deterioration in cow health as a correlated response to selection for increased yield.

Texture of butter from cows with different milk fatty acid compositions.

Milk fatty acid composition and textural properties of butter are known to be affected by the cows' diets. We examined the phenotypic variation in milk fatty acid composition among cows fed the same diet to see if the variation was sufficient to produce butter with different textural properties. Ten cows were selected that tested higher (n = 5) or lower (n = 5) in their proportion of milk unsaturated fatty acids. Milk samples were collected a week after testing, and butter was prepared from the individual samples. Milk and butter samples were again analyzed for fatty acid composition. Butter at 5 degrees C was evaluated by a sensory panel for spreadability and by a texture analyzer at both 5 and 23 degrees C for hardness and adhesiveness. Milk and butter samples from cows with a more unsaturated milk fatty acid composition had a lower atherogenic index, and the butter samples were more spreadable, softer, and less adhesive. Thus, phenotypic variation in milk fatty acid composition among cows fed the same diet is sufficient to produce butter with different textural properties.

Effects of including a quantitative trait locus in selection under different waiting plans of young bulls.

The advantage of using the genotype of a quantitative trait locus (QTL) in selection schemes of dairy cattle was quantified using stochastic simulation. Three selection plans were studied. In the first plan, young bulls waited for 3 yr until their sisters completed a lactation and then were evaluated and selected based on an animal model. In a second plan, young bulls waited for 5 yr until their daughters completed a lactation. An intermediate 4-yr waiting plan was also studied. Simulation was for 16 yr with overlapping generations. Population and model parameters were proportional to the U.S. Holstein population. The advantage of using a QTL was quantified as the percentage of superiority of QTL-assisted over QTL-free selection using cumulative genetic response. Percentage of superiority was reported for four selection pathways: active sires, young bulls, bull dams, and first lactation cows. A general trend was observed: low superiority in early years of selection that increased to a plateau in later years and then decreased. The superiority of the QTL information was greatest in the 3-yr waiting plan and least in the 4-yr waiting plan. Superiority at plateau for selection pathways ranged from 16 to 26% for the 3-yr waiting plan, from 3 to 12% for the 4-yr waiting plan, and from 5 to 13% for the 5-yr waiting plan. The contribution to selection response attributed to the QTL and the polygenes was quantified. The rate at which the favorable allele approached fixation and the accuracy of predicting breeding values on the percentage of superiority were studied.

Superiority of QTL-assisted selection in dairy cattle breeding schemes.

The superiority of selection schemes employing information about a known quantitative trait locus (QTL) over conventional schemes is examined for dairy cattle breeding schemes. Stochastic simulation of a dairy cattle population with selection practices, structures, and parameters similar to the US Holstein population was implemented. Additive genetic effects were estimated by an animal model. Two schemes were compared: a QTL-assisted selection scheme in which the genotype of a known QTL was accounted for in the animal model as a fixed factor, and a QTL-free selection scheme in which the QTL was simulated but was not fit separately in the animal model. Under the QTL-assisted selection scheme, all animals in the mixed model were assumed to be genotyped for the QTL. The effect of using QTL information on the genetic response, the frequency of the favorable QTL allele, and the accuracy of evaluation were examined. Moreover, the effect was studied in four distinct paths of selection: active sires, proven young bulls, bull dams, and first-lactation cows. Average superiority values of 4.6, 7.6, 11.7, and 1.1% for genetic response were observed over 16 yr of selection for active sires, young bulls, bull dams, and first-lactation cows, respectively. Frequency of the favorable QTL allele changed faster in bull dams than males, and was the slowest in first-lactation cows. Finally, accuracy of evaluation under the QTL-assisted selection scheme was higher than under the QTL-free selection scheme. Young bulls ofthe QTL-assisted selection scheme on average had 0.049 higher accuracy, and first-lactation cows had on average 0.185 higher accuracy than corresponding animals of the QTL-free selection scheme.

Genetic control of disease resistance and immunoresponsiveness.

A great deal of evidence points to substantial genetic control over at least some of the immune responses, although genetic parameters for clinical disease have been less favorable. The past two decades have illustrated that single genes with a large impact on food animal health do exist and can be used to improve the health of domestic populations. The current focus on molecular genetics within food animal species will likely unveil numerous other examples of single genes with large effects, although the use of animals possessing favorable genotypes for disease resistance may represent a compromise in selection for increased production of raw product. Moreover, it is also clear that genetic control over the immune system is not limited to a few genes but is more likely influenced by many genes, each with small effects. The use of this information in animal improvement programs is not straightforward because of factors complicating the identification of superior individuals within the population. The scarcity of information dealing with phenotypic and genetic relationships between measures of disease resistance and aspects of immune response complicates the situation even further. Despite these potential hurdles, the potential for permanent improvement of disease resistance within food animal species in the future is tantalizing and merits intensified future study.

A rapid method for computing the inverse of the gametic covariance matrix between relatives for a marked quantitative trait locus.

The inverse of the gametic covariance matrix between relatives, G(-1), for a marked quantitative trait locus (QTL) is required in best linear unbiased prediction (BLUP) of breeding values if marker data are available on a QTL. A rapid method for computing the inverse of a gametic relationship matrix for a marked QTL without building G itself is presented. The algorithm is particularly useful due to the approach taken in computing inbreeding coefficients by having to compute only few elements of G. Numerical techniques for determining, storing, and computing the required elements of G and the nonzero elements of the inverse are discussed. We show that the subset of G required for computing the inbreeding coefficients and hence the inverse is a tiny proportion of the whole matrix and can be easily stored in computer memory using sparse matrix storage techniques. We also introduce an algorithm to determine the maximum set of nonzero elements that can be found in G(-1) and a strategy to efficiently store and access them. Finally, we demonstrate that the inverse can be efficiently built using the present techniques for very large and inbred populations.

Heritability of teat-end shape and the relationship of teat-end shape with somatic cell score for an experimental herd of cows.

Teat-end shapes were categorized for 1443 Holstein cows with 3582 lactations, in the Iowa State University herd at Ankeny, approximately 40 d postpartum between 1970 and 1995. Frequencies of teat-end shapes were as follows: round, 52.2%; prolapsed, 0.8%; flat, 14.2%; plate, 3.4%; funnel, 8.9%; and mixed, 20.5%. Cows were coded as mixed if all four teat-end shapes were not the same. Heritability estimates were obtained with an animal model with pedigrees traced back to registration numbers roughly conforming to birth year 1955. The heritability estimates for first, second, and third and later lactations were 34, 21, and 13%, respectively. Lactation averages for linear somatic cell scores were adjusted for days in milk, and month and age at calving, and were available for 255 cows with 431 lactations categorized from 1992 to 1995. Frequencies of teat-end shape on the 255 cows were as follows: round, 58%; prolapsed, 3%; flat, 11%; plate, 2%; funnel, 6%; and mixed, 20%. Least-squares means of somatic cell score for categories of teat-end shape were computed from a mixed model that included year, parity, and teat-end shape as fixed effects and cow as a random effect. Teat-end shape did not significantly affect somatic cell score.

Sequence heteroplasmy of D-loop and rRNA coding regions in mitochondrial DNA from Holstein cows of independent maternal lineages.

A mitochondrial DNA (mtDNA) fragment containing the D-loop, phenylalanine tRNA, valine tRNA, and 12S and 16 rRNA genes was cloned and sequenced from 36 cows of 18 maternal lineages to identify the polymorphic sites within those regions and to detect the existence of heteroplasmic mtDNA in cows. Seventeen variable sites were observed within the D-loop and rRNA coding regions of bovine mtDNA within a 2.5-kb span. The hypervariable sites in the D-loop and rRNA coding regions were identified at nucleotide positions 169, 216, and 1594. Heteroplasmic mtDNA (variable mtDNA within a tissue) existed extensively in cows and was detected within the above regions in 11 of 36 cows sequenced. The insertion, deletion, and nucleotide transversion polymorphisms were found only in homopolymer regions. Heteroplasmy was observed frequently and seemingly is persistent in cattle. Though heteroplasmy was demonstrated, most lineages and mtDNA sites showed no heteroplasmy.

Direct and correlated responses to selection for milk yield: results and conclusions of regional project NC-2, "improvement of dairy cattle through breeding, with emphasis on selection". NC-2 Technical Committee.

Measurement of direct and correlated responses to single-trait selection for milk yield was the major objective of regional project NC-2. The NC-2 Technical Committee included representatives from Alaska, Illinois, Indiana, Iowa, Kansas, Michigan, Minnesota, Nebraska, South Dakota, Wisconsin, and the USDA. All representatives, except Illinois, Kansas and Nebraska, maintained a selection line formed by using AI sires selected for high estimated transmitting abilities for milk and a second line that served as some type of a control. Stations varied in criteria for selection of bulls for control lines. Farms were managed similarly, including feeding and management of selection and control lines as one herd, random mating within line, and restricted culling policies. Selection for milk yield effectively increased milk production. All selection lines increased milk and net income per lactation more than control lines. Realized gains matched or exceeded gains expected from estimates of breeding values. Yields of milk components increased, but component percentages decreased appreciably for selection lines. Reproduction of nulliparous animals was not affected, but days open for lactating selection cows increased in some of the individual projects. Selected cows tended to have larger health costs, specifically for mammary treatment. Udder and conformation traits did not deteriorate for selection lines, although control lines with selection of sires on genetic evaluations for type received higher type scores. There should be few reservations about undesirable responses correlated with selection for milk yield.

Associations among individual proteins and fatty acids in bovine milk as determined by correlations and factor analyses.

Associations among quantities and concentrations of individual milk proteins and fatty acids were determined in individual milk samples from 233 Holstein cows. Correlation coefficients among the six major proteins and the eleven major fatty acids in bovine milk were grouped hierarchically. Factor analyses grouped the milk components into seven families: fatty acids 4:0-6:0, 6:0-16:0, 16:0, 18:0, 16:1 plus 18:1 plus 18:2, all milk proteins and beta-lactoglobulin alone. Correlation coefficients and groupings by factor analyses coincided with shared pathways of synthesis or genetic origins of milk proteins and fatty acids because they are the basis of the correlation coefficients. Hence, the results from correlations and factor analyses could be used to develop hypotheses for the synthesis of milk components and other coordinately regulated physiological processes.

Association of growth hormone loci with milk yield traits in Holstein bulls.

A pedigree analysis was used to investigate the association of bovine growth hormone loci with milk production traits of Holstein cattle. Holstein bulls were typed for three bovine growth hormone loci located in exon V, intron C, and the 3' region of the gene. Phenotypic data were daughter yield deviations for milk, fat, and protein yields and for fat and protein percentages. Analysis of linkage across families was applied to the data using one or two bovine growth hormone loci as markers linked to a putative biallelic quantitative trait locus. Estimated parameters were allele frequency, genotypic means, within-genotype standard deviation of a putative quantitative trait locus, and recombination fraction between the markers and the quantitative trait locus. Parameters were estimated by maximum likelihood techniques. The estimated frequency of the quantitative trait locus allele that decreased the value of the phenotype ranged from 0.1 for milk yield to 0.6 for protein yield. The estimated effect of an allele substitution at the quantitative trait locus, given in phenotypic standard deviation units, ranged from 0.75 for fat percentage to 1.6 for milk yield. The standard deviation within genotype ranged from 0.67 for fat yield to 0.87 for milk yield. The estimated recombination fraction was close to zero for protein percentage, indicating physical linkage between a quantitative trait locus affecting the trait and the bovine growth hormone loci.

Pregnancy-associated glycoprotein and decreased polymorphonuclear leukocyte function in early post-partum dairy cows.

Phagocytosis and oxidative burst activity of polymorphonuclear neutrophil leukocytes (PMN) isolated from blood and pregnancy-associated glycoprotein (bPAG) concentrations in plasma were evaluated in two longitudinal studies in dairy cows from 3 weeks before until 5 weeks after calving, carried out in the United States and in Europe. Ingestion of Staphylococcus aureus by blood PMN increased during the first week after calving and normalised 3 weeks post-partum. Phagocytosis of Escherichia coli did not change in the early post-partum period. In both studies, a significant decrease in oxidative burst activity of PMN was observed between 1 and 3 weeks after calving. In all cows, a very significant increase in plasma bPAG concentration was found between 1 week before and 2 weeks after calving. The peak of bPAG concentration in plasma immediately preceded the alterations of blood PMN functions. These results suggest that bPAG may be associated with inhibition of PMN function of dairy cows during the early post-partum period.

Effect of milk protein genotypes on milk protein composition and its genetic parameter estimates.

The effects of kappa-casein (CN) and beta-lactoglobulin (LG) genotypes on milk protein concentration and composition were estimated for the US Holstein-Friesian population using a single-trait, mixed, linear animal model on 592 individual milk samples from 233 cows. Both milk protein genotypes had no statistically significant effect on the total milk protein concentration; however, substitution of the kappa-CN A allele additively increased the proportion of kappa-CN, and substitution of the beta-LG B allele additively increased the proportion of beta-LG in total milk protein. In response, proportions of the other milk proteins, mainly alpha S1-CN, were decreased. For proportions of alpha S1-CN, kappa-CN, and beta-LG in total milk protein, kappa-CN and beta-LG genotypes explain more than 50 and 25% of the heritability and repeatability estimates, respectively. We concluded that kappa-CN and beta-LG genotypes affect the phenotypic and genetic variation of milk protein composition but do not significantly affect milk protein concentration. A possible explanation for our conclusion is that altered gene sequences in the promoter region of kappa-CN B and beta-LG A, linked closely to the respective genotypes, favor the transcription or translation of their own protein at the expense of the synthesis of other milk proteins, in particular of alpha S1-CN.

Genetic association between parameters of inmate immunity and measures of mastitis in periparturient Holstein cattle.

Relationships between genetic measures of mastitis (somatic cell score, score for clinical mastitis, and scores for IMI with major or minor pathogens) and immunological parameters (physiological and molecular markers) were examined for periparturient Holstein cows. Physiological markers included 11 in vitro immunological assays. Molecular markers included the second exon of the DRB3 locus of the bovine major histocompatibility complex, the IgG2 isotype genotype, and the CD18 genotype (the locus responsible for bovine leukocyte adhesion deficiency). A gene substitution model was used to estimate the additive genetic effects of alleles of the three molecular markers on estimated breeding value (EBV) for mastitis measures. Pearson correlation coefficients between EBV for immunological assays and EBV for mastitis measures were computed. Molecular markers explained up to 40% of the variation in EBV for measures of mastitis. The presence of allele DRB3.2*16 was associated with higher EBV for SCS. Allele DRB3.2*8 was associated with increased EBV for clinical mastitis, as was the IgG2b allele and the normal CD18 allele. Alleles DRB3.2*11, *23, IgG2a, and the recessive allele for bovine leukocyte adhesion deficiency were associated with decreased clinical mastitis. A positive genetic association was found between allele DRB3.2*24 and EBV for IMI by major pathogens and between DRB3.2*3 and IMI by minor pathogens. Several correlations between EBV for immunological assays and EBV for mastitis measures were significantly different from 0. Cows with low EBV for SCS tended to have neutrophils that had greater functional ability at maximal immunosuppression, low serum IgG1, and high numbers of circulating mononuclear cells. Immunological parameters, including physiological and molecular markers, are useful aids to understand the genetics of resistance to mastitis.