Genome-wide associations for heat stress response suggest potential candidate genes underlying milk fatty acid composition in dairy cattle.

Contents of milk fatty acids (FA) display remarkable alterations along climatic gradients. Detecting candidate genes underlying such alterations might be beneficial for the exploration of climate sensitivity in dairy cattle. Consequently, we aimed on the definition of FA heat stress indicators, considering FA breeding values in response to temperature-humidity index (THI) alterations. Indicators were used in GWAS, in ongoing gene annotations and for the estimation of chromosome-wide variance components. The phenotypic data set consisted of 39,600 test-day milk FA records from 5,757 first-lactation Holstein dairy cows kept in 16 large-scale German cooperator herds. The FA traits were C18:0, polyunsaturated fatty acids (PUFA), saturated fatty acids (SFA), and unsaturated fatty acids (UFA). After genotype quality control, 40,523 SNP markers from 3,266 cows and 930 sires were considered. Meteorological data from the weather station in closest herd distance were used for the calculation of maximum hourly daily THI, which were allocated to 10 different THI classes. The same FA from 3 stages of lactation were considered as different, but genetically correlated traits. Consequently, a 3-trait reaction norm model was used to estimate genetic parameters and breeding values for FA along THI classes, considering either pedigree (A) or genomic (G) relationship matrices. De-regressed proofs and genomic estimated breeding values at the intermediate THI class 5 and at the extreme THI class 10 were used as pseudophenotypes in ongoing genomic analyses for thermoneutral (TNC) and heat stress conditions (HSC), respectively. The differences in de-regressed proofs and in genomic estimated breeding values from both THI classes were pseudophenotypes for heat stress response (HSR). Genetic correlations between the same FA under TNC and HSC were smallest in the first lactation stage and ranged from 0.20 for PUFA to 0.87 for SFA when modeling with the A matrix, and from 0.35 for UFA to 0.86 for SFA when modeling with the G matrix. In the first lactation stage, larger additive genetic variances under HSC compared with TNC indicate climate sensitivity for C18:0, PUFA, and UFA. Climate sensitivity was also reflected by pronounced chromosome-wide genetic variances for HSR of PUFA and UFA in the first stage of lactation. For all FA under TNC, HSC, and HSR, quite large genetic variance proportions were explained by BTA14. In GWAS, 30 SNP (within or close to 38 potential candidate genes) overlapped for HSR of the different FA. One unique potential candidate gene (AMFR) was detected for HSR of PUFA, 15 for HSR of SFA (ADGRB1, DENND3, DUSP16, EFR3A, EMP1, ENSBTAG00000003838, EPS8, MGP, PIK3C2G, STYK1, TMEM71, GSG1, SMARCE1, CCDC57, and FASN) and 3 for HSR of UFA (ENSBTAG00000048091, PAEP, and EPPK1). The identified unique genes play key roles in milk FA synthesis and are associated with disease resistance in dairy cattle. The results suggest consideration of FA in combination with climatic responses when inferring genetic mechanisms of heat stress in dairy cows.

Climate sensitivity of milk production traits and milk fatty acids in genotyped Holstein dairy cows.

The aim of this study was the evaluation of climate sensitivity via genomic reaction norm models [i.e., to infer cow milk production and milk fatty acid (FA) responses on temperature-humidity index (THI) alterations]. Test-day milk traits were recorded between 2010 and 2016 from 5,257 first-lactation genotyped Holstein dairy cows. The cows were kept in 16 large-scale cooperator herds, being daughters of 344 genotyped sires. The longitudinal data consisted of 47,789 test-day records for the production traits milk yield (MY), fat yield (FY), and protein yield (PY), and of 20,742 test-day records for 6 FA including C16:0, C18:0, saturated fatty acids (SFA), unsaturated fatty acids (UFA), monounsaturated fatty acids (MUFA), and polyunsaturated fatty acids (PUFA). After quality control of the genotypic data, 41,057 SNP markers remained for genomic analyses. Meteorological data from the weather station in closest herd distance were used for the calculation of maximum hourly daily THI. Genomic reaction norm models were applied to estimate genetic parameters in a single-step approach for production traits and FA in dependency of THI at different lactation stages, and to evaluate the model stability. In a first evaluation strategy (New_sire), all phenotypic records from daughters of genotyped sires born after 2010 were masked, to mimic a validation population. In the second strategy (New_env), only daughter records of the new sires recorded in the most extreme THI classes were masked, aiming at predicting sire genomic estimated breeding values (GEBV) under heat stress conditions. Model stability was the correlation between GEBV of the new sires in the reduced data set with respective GEBV estimated from all phenotypic data. Among all test-day production traits, PY responded as the most sensitive to heat stress. As observed for the remaining production traits, genetic variances were quite stable across THI, but genetic correlations between PY from temperate climates with PY from extreme THI classes dropped to 0.68. Genetic variances in dependency of THI were very similar for C16:0 and SFA, indicating marginal climatic sensitivity. In the early lactation stage, genetic variances for C18:0, MUFA, PUFA, and UFA were significantly larger in the extreme THI classes compared with the estimates under thermoneutral conditions. For C18:0 and MUFA, PUFA, and UFA in the middle THI classes, genetic correlations in same traits from the early and the later lactation stages were lower than 0.50, indicating strong days in milk influence. Interestingly, within lactation stages, genetic correlations for C18:0 and UFA recorded at low and high THI were quite large, indicating similar genetic mechanisms under stress conditions. The model stability was improved when applying the New_env instead of New_sire strategy, especially for FA in the first stage of lactation. Results indicate moderately accurate genomic predictions for milk traits in extreme THI classes when considering phenotypic data from a broad range of remaining THI. Phenotypically, thermal stress conditions contributed to an increase of UFA, suggesting value as a heat stress biomarker. Furthermore, the quite large genetic variances for UFA at high THI suggest the consideration of UFA in selection strategies for improved heat stress resistance.

Prediction accuracies and genetic parameters for test-day traits from genomic and pedigree-based random regression models with or without heat stress interactions.

The aim of this study was to compare genetic (co)variance components and prediction accuracies of breeding values from genomic random regression models (gRRM) and pedigree-based random regression models (pRRM), both defined with or without an additional environmental gradient. The used gradient was a temperature-humidity index (THI), considered in statistical models to investigate possible genotype by environment (G×E) interactions. Data included 106,505 test-day records for milk yield (MY) and 106,274 test-day records for somatic cell score (SCS) from 12,331 genotyped Holstein Friesian daughters of 522 genotyped sires. After single nucleotide polymorphism quality control, all genotyped animals had 40,468 single nucleotide polymorphism markers. Test-day traits from recording years 2010 to 2015 were merged with temperature and humidity data from the nearest weather station. In this regard, 58 large-scale farms from the German federal states of Berlin-Brandenburg and Mecklenburg-West Pomerania were allocated to 31 weather stations. For models with a THI gradient, additive genetic variances and heritabilities for MY showed larger fluctuations in dependency of DIM and THI than for SCS. For both traits, heritabilities were smaller from the gRRM compared with estimates from the pRRM. Milk yield showed considerably larger G×E interactions than SCS. In genomic models including a THI gradient, genetic correlations between different DIM × THI combinations ranged from 0.26 to 0.94 for MY. For SCS, the lowest genetic correlation was 0.78, estimated between SCS from the last DIM class and the highest THI class. In addition, for THI × THI combinations, genetic correlations were smaller for MY compared with SCS. A 5-fold cross-validation was used to assess prediction accuracies from 4 different models. The 4 different models were gRRM and pRRM, both modeled with or without G×E interactions. Prediction accuracy was the correlation between breeding values for the prediction data set (i.e., excluding the phenotypic records from this data set) with respective breeding values considering all phenotypic information. Prediction accuracies for sires and for their daughters were largest for the gRRM considering G×E interactions. Such modeling with 2 covariates, DIM and THI, also allowed accurate predictions of genetic values at specific DIM. In comparison with a pRRM, the effect of a gRRM with G×E interactions on gain in prediction accuracies was stronger for daughters than for sires. In conclusion, we found stronger effect of THI alterations on genetic parameter estimates for MY than for SCS. Hence, gRRM considering THI especially contributed to gain in prediction accuracies for MY.

Genomic breeding values, SNP effects and gene identification for disease traits in cow training sets.

Holstein Friesian cow training sets were created according to disease incidences. The different datasets were used to investigate the impact of random forest (RF) and genomic BLUP (GBLUP) methodology on genomic prediction accuracies. In addition, for further verifications of some specific scenarios, single-step genomic BLUP was applied. Disease traits included the overall trait categories of (i) claw disorders, (ii) clinical mastitis and (iii) infertility from 80 741 first lactation Holstein cows kept in 58 large-scale herds. A subset of 6744 cows was genotyped (50K SNP panel). Response variables for all scenarios were de-regressed proofs (DRPs) and pre-corrected phenotypes (PCPs). Initially, all sick cows were allocated to the testing set, and healthy cows represented the training set. For the ongoing cow allocation schemes, the number of sick cows in the training set increased stepwise by moving 10% of the sick cows from the testing to the training set in each step. The size of training and testing sets was kept constant by replacing the same number of cows in the testing set with (randomly selected) healthy cows from the training set. For both the RF and GBLUP methods, prediction accuracies were larger for DRPs compared to PCPs. For PCPs as a response variable, the largest prediction accuracies were observed when the disease incidences in training sets reflected the disease incidence in the whole population. A further increase in prediction accuracies for some selected cow allocation schemes (i.e. larger prediction accuracies compared to corresponding scenarios with RF or GBLUB) was achieved via single-step GBLUP applications. Correlations between genome-wide association study SNP effects and RF importance criteria for single SNPs were in a moderate range, from 0.42 to 0.57, when considering SNPs from all chromosomes or from specific chromosome segments. RF identified significant SNPs close to potential positional candidate genes: GAS1, GPAT3 and CYP2R1 for clinical mastitis; SPINK5 and SLC26A2 for laminitis; and FGF12 for endometritis.