K.A. Thompson-Crispi

Bovine Mastitis: Frontiers in Immunogenetics

Mastitis is one of the most prevalent and costly diseases in the dairy industry with losses attributable to reduced milk production, discarded milk, early culling, veterinary services, and labor costs. Typically, mastitis is an inflammation of the mammary gland most often, but not limited to, bacterial infection, and is characterized by the movement of leukocytes and serum proteins from the blood to the site of infection. It contributes to compromised milk quality and the potential spread of antimicrobial resistance if antibiotic treatment is not astutely applied. Despite the implementation of management practises and genetic selection approaches, bovine mastitis control continues to be inadequate. However, some novel genetic strategies have recently been demonstrated to reduce mastitis incidence by taking advantage of a cow’s natural ability to make appropriate immune responses against invading pathogens. Specifically, dairy cattle with enhanced and balanced immune responses have a lower occurrence of disease, including mastitis, and they can be identified and selected for using the high immune response (HIR) technology. Enhanced immune responsiveness is also associated with improved response to vaccination, increased milk, and colostrum quality. Since immunity is an important fitness trait, beneficial associations with longevity and reproduction are also often noted. This review highlights the genetic regulation of the bovine immune system and its vital contributions to disease resistance. Genetic selection approaches currently used in the dairy industry to reduce the incidence of disease are reviewed, including the HIR technology, genomics to improve disease resistance or immune response, as well as the Immunity+™ sire line. Improving the overall immune responsiveness of cattle is expected to provide superior disease resistance, increasing animal welfare and food quality while maintaining favorable production levels to feed a growing population.

Genetic parameters of adaptive immune response traits in Canadian Holsteins

The objectives of this study were to estimate genetic parameters of cell-mediated (CMIR) and antibody-mediated (AMIR) immune response (IR) traits of Holstein cattle on a national scale and to associate estimated breeding values of CMIR, AMIR, and overall IR with routinely evaluated traits in Canada. In collaboration with the Canadian Bovine Mastitis Research Network, 445 Holstein cows from 42 herds across Canada were immunized to measure delayed-type hypersensitivity as an indicator of CMIR and serum antibody for AMIR to putative type 1 and type 2 test antigens, respectively. Primary (d 14) and secondary (d 21) AMIR were measured for both IgG1 and IgG2. A series of uni- and bivariate linear animal models were used to estimate genetic parameters and breeding values for CMIR and the 4 AMIR traits. The models included the fixed effects of parity and stage of lactation and the random effects of herd-technician, animal, and residual. Heritability of CMIR was 0.19 (SE=0.10) and for AMIR traits ranged from 0.16 to 0.41 (SE=0.09-0.11) depending on time and antibody isotype. The genetic correlations between CMIR and AMIR were negative and ranged from -0.13 to -0.45 (SE=0.32-0.46). The results indicate adaptive immune response traits are moderately heritable and provide a potential for genetic selection. The negative genetic correlations between CMIR and AMIR indicate the importance of considering both traits in breeding for overall disease resistance. Significant beneficial associations between the reproductive traits number of services and first service to conception were found, which may indicate these reproductive traits would improve with genetic selection for enhanced immune response.

Short communication: Association of disease incidence and adaptive immune response in Holstein dairy cows

The objective of this study was to use previously calculated estimated breeding values for cell- (CMIR) and antibody-mediated immune responses (AMIR) to determine associations between immune response (IR) and economically important diseases of dairy cattle. In total, 699 Holsteins were classified as high, average, or low for CMIR, AMIR, and overall IR (combined CMIR and AMIR), and associations with mastitis, metritis, ketosis, displaced abomasums, and retained fetal membranes were determined. The incidence of mastitis was higher among average cows as compared with cows classified as high AMIR [odds ratio (OR)=2.5], high CMIR (OR=1.8), or high IR (OR=1.8). Low-CMIR cows had a higher incidence of metritis (OR=11.3) and low-IR cows had a higher incidence of displaced abomasum (OR=4.1) and retained fetal membrane (OR=2.8) than did average responders. Results of this study show that cows classified as high immune responders have lower occurrence of disease, suggesting that breeding cattle for enhanced IR may be a feasible approach to decrease the incidence of infectious and metabolic diseases in the dairy industry.

Incidence Rates of Clinical Mastitis among Canadian Holsteins Classified as High, Average, or Low Immune Responders

ABSTRACT The objective of this study was to compare the incidence rate of clinical mastitis (IRCM) between cows classified as high, average, or low for antibody-mediated immune responses (AMIR) and cell-mediated immune responses (CMIR). In collaboration with the Canadian Bovine Mastitis Research Network, 458 lactating Holsteins from 41 herds were immunized with a type 1 and a type 2 test antigen to stimulate adaptive immune responses. A delayed-type hypersensitivity test to the type 1 test antigen was used as an indicator of CMIR, and serum antibody of the IgG1 isotype to the type 2 test antigen was used for AMIR determination. By using estimated breeding values for these traits, cows were classified as high, average, or low responders. The IRCM was calculated as the number of cases of mastitis experienced over the total time at risk throughout the 2-year study period. High-AMIR cows had an IRCM of 17.1 cases per 100 cow-years, which was significantly lower than average and low responders, with 27.9 and 30.7 cases per 100 cow-years, respectively. Low-AMIR cows tended to have the most severe mastitis. No differences in the IRCM were noted when cows were classified based on CMIR, likely due to the extracellular nature of mastitis-causing pathogens. The results of this study demonstrate the desirability of breeding dairy cattle for enhanced immune responses to decrease the incidence and severity of mastitis in the Canadian dairy industry.

A genome-wide association study of immune response traits in Canadian Holstein cattle

BackgroundBreeding for enhanced immune response (IR) has been suggested as a tool to improve inherent animal health. Dairy cows with superior antibody-mediated (AMIR) and cell-mediated immune responses (CMIR) have been demonstrated to have a lower occurrence of many diseases including mastitis. Adaptive immune response traits are heritable, and it is, therefore, possible to breed for improved IR, decreasing the occurrence of disease. The objective of this study was to perform genome-wide association studies to determine differences in genetic profiles among Holstein cows classified as High or Low for AMIR and CMIR. From a total of 680 cows with immune response phenotypes, 163 cows for AMIR (81 High and 82 Low) and 140 for CMIR (75 High and 65 Low) were selectively genotyped using the Illumina Bovine SNP50 BeadChip. Results were validated using an unrelated population of 164 Holstein bulls IR phenotyped for AMIR and 146 for CMIR.ResultsA generalized quasi likelihood score method was used to determine single nucleotide polymorphisms (SNP) and chromosomal regions associated with immune response. After applying a 5% chromosomal false discovery rate, 186 SNPs were significantly associated with AMIR. The majority (93%) of significant markers were on chromosome 23, with a similar peak found in the bull population. For CMIR, 21 SNP markers remained significant. Candidate genes within 250,000 base pairs of significant SNPs were identified to determine biological pathways associated with AMIR and CMIR. Various pathways were identified, including the antigen processing and presentation pathway, important in host defense. Candidate genes included those within the bovine Major Histocompatability Complex such as BoLA-DQ, BoLA-DR and the non-classical BoLA-NC1 for AMIR and BoLA-DQ for CMIR, the complement system including C2 and C4 for AMIR and C1q for CMIR, and cytokines including IL-17A, IL17F for AMIR and IL-17RA for CMIR and tumor necrosis factor for both AMIR and CMIR. Additional genes associated with CMIR included galectins 1, 2 and 3, BCL2 and β-defensin.ConclusionsThe significant genetic variation associated with AMIR and CMIR in this study may imply feasibility to include immune response in genomic breeding indices as an approach to improve inherent animal health.

Genetic parameters for natural antibodies and associations with specific antibody and mastitis in Canadian Holsteins.

The objectives of this study were to evaluate (1) natural antibodies (NAb) in Canadian Holstein cows, (2) genetic parameters and associations between NAb and specific antibody (SpAb), and (3) the association of NAb with clinical mastitis and differences in incidence rates of clinical mastitis (IRCM) among cows classified as high, average, or low responders for NAb. Natural antibodies (IgG and IgM) to keyhole limpet hemocyanin and SpAb to a type 2 test antigen were measured on 451 Holsteins from 41 herds across Canada. A series of uni- and tri-variate linear animal models were used to estimate genetic parameters and breeding values for NAb and SpAb. The models included the fixed effects of parity and stage of lactation and the random effects of herd-technician, animal, and residual. Using estimated breeding values for NAb, cows were classified as high, average, or low responders and phenotypic associations with the IRCM were investigated and a logistic regression performed. The estimated heritability was 0.27 for SpAb, and was 0.32 and 0.18 for NAb of the IgG and IgM isotypes, respectively. No significant genetic correlations were found between SpAb and NAb. Although no significant differences in the IRCM were found when cows were classified based on NAb IgG, cows classified as high responders for NAb IgM tended to have a lower IRCM compared with other cows. Immunoglobulin-M was associated with a decreased risk of clinical mastitis (odds ratio=0.958). Results of this study suggest the potential to use NAb IgM as an additional tool to select for disease resistance in cattle, but results need to be validated with a larger sample size.

Short communication: Variation of total immunoglobulin G and β-lactoglobulin concentrations in colostrum and milk from Canadian Holsteins classified as high, average, or low immune responders

The objective of this study was to evaluate IgG and β-lactoglobulin (β-LG) concentrations in colostrum and milk of Canadian Holsteins (n=108) classified as high (H), average (A), or low (L) for antibody-mediated (AMIR) or cell-mediated immune responses (CMIR) based on estimated breeding values. It was hypothesized that H-AMIR and H-CMIR cows produce colostrum (first milking) and milk (d 5 postcalving) with higher concentrations of IgG and β-LG. Data for IgG and β-LG in colostrum and milk were analyzed independently using mixed linear models. Least squares means were compared using Tukeys test. Cows classified as H-AMIR had higher IgG and β-LG concentrations in colostrum compared with A- and L-AMIR cows; 84% of H-AMIR, 69% of A-AMIR, and 68% of L-AMIR cows had over 5,000 mg/dL IgG in colostrum. No differences in IgG and β-LG concentrations in colostrum were noted among cows ranked on CMIR or in milk of cows ranked on AMIR. β-Lactoglobulin and IgG concentrations were positively correlated in colostrum. Breeding cows for H-AMIR status may reduce failure of passive transfer of IgG in their calves; β-LG may play a role in bovine immune defenses. Colostrum from H-AMIR cows may serve as a more economical feedstock source for manufacturing natural health products.

Short communication: Cytokine profiles from blood mononuclear cells of dairy cows classified with divergent immune response phenotypes

Genetic selection for enhanced immune response has been shown to decrease disease occurrence in dairy cattle. Cows can be classified as high (H), average, or low responders based on antibody-mediated immune response (AMIR), predominated by type-2 cytokine production, and cell-mediated immune response (CMIR) through estimated breeding values for these traits. The purpose of this study was to identify in vitro tests that correlate with in vivo immune response phenotyping in dairy cattle. Blood mononuclear cells (BMC) isolated from cows classified as H-AMIR and H-CMIR through estimated breeding values for immune response traits were stimulated with concanavalin A (ConA; Sigma Aldrich, St. Louis, MO) and gene expression, cytokine production, and cell proliferation was determined at multiple time points. A repeated measures model, which included the effects of immune response group, parity, and stage of lactation, was used to compare differences between immune response phenotype groups. The H-AMIR cows produced more IL-4 protein than H-CMIR cows at 48 h; however, no difference in gene expression of type-2 transcription factor GATA3 or IL4 was noted. The BMC from H-CMIR cows had increased production of IFN-γ protein at 48, 72, and 96 h compared with H-AMIR animals. Further, H-CMIR cows had increased expression of the IFNG gene at 16, 24, and 48 h post-treatment with ConA, although expression of the type-1 transcription factor gene TBX21 did not differ between immune response groups. Although proliferation of BMC increased from 24 to 72 h after ConA stimulation, no differences were found between the immune response groups. Overall, stimulation of H-AMIR and H-CMIR bovine BMC with ConA resulted in distinct cytokine production profiles according to genetically defined groups. These distinct cytokine profiles could be used to define disease resistance phenotypes in dairy cows according to stimulation in vitro; however, other immune response phenotypes should be assessed.

Short communication: Variation in production parameters among Canadian Holstein cows classified as high, average, and low immune responders

Dairy cattle evaluated for immune responses and identified as high responders are known to have a lower occurrence of economically important diseases, including mastitis, metritis, ketosis, and retained placenta. These high immune responders have also been shown to make more antibody following vaccination and to have improved milk and colostrum quality. Therefore, breeding for improved immune response is expected to have several benefits in the dairy industry. However, a concern of such an approach to improve animal health is the potential cost of lost production due to an allocation of host resources to mount a robust immune response. The objective of this study was to evaluate early- and late-lactation production parameters in cattle classified as having high, average, or low estimated breeding values (EBV) for cell-mediated (CMIR), antibody-mediated (AMIR), and overall immune responses. A total of 561 cows from 6 herds were phenotyped for immune response and ranked based on EBV for CMIR and AMIR. A linear animal model was used to evaluate differences in milk, fat, and protein yields among immune response groups, and a regression analysis was conducted based on immune response EBV. Overall, no difference in production parameters was found based on immune response rank; however, some positive relationships with immune response EBV were found, suggesting that breeding for enhanced immune responsiveness as a prophylactic approach to improve animal health would not come at the cost of lost production.

A genome-wide association study for natural antibodies measured in blood of Canadian Holstein cows

BackgroundNatural antibodies (NAb) are an important component of the innate immune system, and fight infections as a part of the first line defence. NAb are poly-reactive and can respond non-specifically to antigens. Therefore, NAb may be a key trait when evaluating an animal’s potential natural disease resistance. Variation in NAb is caused by both genetic and environmental factors. In this study genetic parameters of NAb were estimated and a genome-wide association study (GWAS) was performed to gain further understanding on the genes that are responsible for the observed genetic variation of NAb in Canadian Holsteins.ResultsIn total, blood samples of 1327 cows from 64 farms were studied. NAb binding to keyhole limpet hemocyanin (KLH) were determined via indirect ELISA. Immunoglobulin (Ig) isotypes, IgG and IgM, were evaluated. From the sample population, 925 cows were genotyped for 45,187 markers and each individual marker was tested to detect genetic variation in NAb levels. The relationships among animals was accounted for with genomic relationship. Results show heritabilities of 0.27 ± 0.064 (IgG) and 0.31 ± 0.065 (IgM). In total, 23 SNPs were found to be associated with IgG, but no SNPs were associated with IgM (FDR p-value < 0.05). The significant SNPs were located on autosomal chromosomes 1, 20 and 21 of the cow genome. Functional annotation analysis of the positional candidate genes revealed two sets of genes with biologically relevant functions related to NAb. In one set, seven genes with crucial roles in the production of antibody in B cells were associated with the trafficking of vesicles inside the cells between organelles. In the second set, two genes among positional candidate genes were associated with isotype class-switching and somatic hypermutation of B cells.ConclusionsThis study demonstrated the possibility of increasing NAb through selective breeding. In addition, the effects of two candidate pathways are proposed for further investigation of NAb production in Holsteins.