Multiple births or twinning in cattle is a naturally occurring reproductive phenomenon. For dairy cattle, twinning is considered a detrimental trait as it can be harmful to cow and calf as well as costly to the producer. The objective of this study was to examine recent US calving records for the Holstein breed to determine a current estimate of heritability for twinning rate along with effects of season and parity. Two models were used in this study: a linear sire model and a binary threshold-logit sire model. Both were mixed models considering fixed effects and random effects. Analyses were conducted using a restricted maximum likelihood method. Heritability estimates were 0.0192 ± 0.0009 and 0.1420 ± 0.0069 for the linear and threshold models, respectively. Repeatabilities from the linear and threshold-logit models were 0.0443 ± 0.0012 and 0.2310 ± 0.0072, respectively. The nonzero estimates of heritability indicate the potential to select against this trait for genetic improvement of Holstein cattle.

Paratuberculosis, or Johne's disease, is a chronic, granulomatous, gastrointestinal tract disease of cattle and other ruminants caused by the bacterium Mycobacterium avium subspecies paratuberculosis (MAP). Control of Johne's disease is based on programs of testing and culling animals positive for infection with MAP and concurrently modifying management to reduce the likelihood of infection. The current study was motivated by the hypothesis that genetic variation in host susceptibility to MAP infection can be dissected and quantifiable associations with genetic markers identified. Two separate GWAS analyses were conducted, the first using 897 genotyped Holstein artificial insemination sires with phenotypes derived from incidence of MAP infection among daughters based on milk ELISA testing records. The second GWAS analysis was a case-control design using US Holstein cows phenotyped for MAP infection by serum ELISA or fecal culture tests. Cases included cows positive for either serum ELISA, fecal culture, or both. Controls consisted of animals negative for all tests conducted. A total of 376 samples (70 cases and 306 controls) from a University of Minnesota Johne's management demonstration project and 184 samples (76 cases and 108 controls) from a Michigan State University study were used. Medium-density (sires) and high-density (cows) genotype data were imputed to full genome sequence for the analyses. Marker-trait associations were analyzed using the single-step (ss)GWAS procedure implemented in the BLUPF90 suite of programs. Evidence of significant genomic contributions for susceptibility to MAP infection were observed on multiple chromosomes. Results were combined across studies in a meta-analysis, and increased support for genomic regions on BTA7 and BTA21 were observed. Gene set enrichment analysis suggested pathways for antigen processing and presentation, antimicrobial peptides and natural killer cell-mediated cytotoxicity are relevant to variation in host susceptibility to MAP infection, among others. Genomic prediction was evaluated using a 5-fold cross-validation, and moderate correlations were observed between genomic breeding value predictions and daughter averages (~0.43 to 0.53) for MAP infection in testing data sets. These results suggest that genomic selection against susceptibility to MAP infection is feasible in Holstein cattle.

Johne's disease in cattle is the result of infection of the small intestine by Mycobacterium avium ssp. paratuberculosis (MAP), leading to an incurable inflammatory bowel disease (Johne's disease or paratuberculosis). The disease is a concern both for its direct cost to dairy producers and for its zoonotic potential. The objective of this study was to estimate the heritability for susceptibility to infection of cattle by MAP using Johne's testing records (ELISA test for presence of antibodies to MAP in milk or blood) from US Holstein cattle from 2009 to 2016. Data sets were edited to include records from herds with 100 or more total records and sires with 50 or more daughters. Data sets were further edited to include (1) only herds with at least 1 positive test, (2) herds with at least 2.5% positive test results, and (3) herds with at least 5% positive test results to examine the effect of data from herds with higher proportions of positive tests, and presumably higher pathogen exposure, on heritability estimates. Two models were used in this study, a linear sire model and a binary threshold-probit sire model. Both were mixed models considering fixed effects of herd and age at test, the latter as a covariate accounting for linear and quadratic effects; random effects included sire and residual. Analyses were conducted using a restricted maximum likelihood method. Heritability estimates (±standard error) from the linear model were 0.041 ± 0.004, 0.050 ± 0.004, and 0.062 ± 0.007 for data from herds with at least 1 positive test, ≥2.5% positive tests, and ≥5% positive tests, respectively. Heritability estimates from the threshold model were 0.157 ± 0.014, 0.174 ± 0.016, and 0.186 ± 0.021 for data from herds with at least 1 positive test, ≥2.5% positive tests, and ≥5% positive tests, respectively. Heritability estimates from the linear model were affected by population incidence for positive tests, in contrast to estimates from the threshold model, likely accounting for the difference in magnitude of heritability estimates between models and suggesting that the threshold model analysis is the better choice. Heritability estimates increased as data were restricted to herds with presumed higher MAP exposure for both linear model and threshold model analyses. These estimates are similar to previous estimates in other dairy cattle populations and suggest the potential for selection to lessen susceptibility to MAP infection.

Bovine twin birth is associated with detriments including increased embryo/fetal losses, malpresentation and dystocia. Incidence of these is lessened in bilateral compared to unilateral twin pregnancy. This study was undertaken to assess use of follicular ablation by aspiration to create bilateral twin pregnancies in females with genetic potential for ~3.5 ovulations per cycle (Trio allele carriers). In Experiment 1, carriers (n=30) and non-carriers (n=10) were synchronized for ovulation and timed artificial insemination (TAI). Follicles (>5 mm) in excess of one per ovary were aspirated ~16h preceding TAI. Follicle count for females with follicles on only one ovary was reduced to two. Blood was sampled 2 weeks post-TAI to assess progesterone (P4) concentrations; embryo count was determined by ultrasound 6 weeks post-TAI. Circulating P4 concentration post-TAI was significantly (p& 0.001) associated with both genotype and subsequent pregnancy status (pregnant non-carriers, 7.06±0.68ng/mL; pregnant carriers, 5.54±0.55ng/mL; non-pregnant non-carriers 5.22±1.05ng/mL; non-pregnant carriers, 3.13±0.42ng/mL). Experiment 2 was undertaken to offset negative effects of follicular aspiration on subsequent P4 concentration observed in Experiment 1. Carriers (n=38) and non-carriers (n=32) were submitted to TAI and follicle ablation as described for Experiment 1. Additionally, accessory corpora lutea (CL) were induced in carriers by administration of human chorionic gonadotropin (carriers) at d6 post-TAI. Consequently, P4 concentration post-TAI was significantly (p& 0.05) associated with subsequent pregnancy status (pregnant, 8.48±0.61ng/mL; non-pregnant, 6.70±0.63 ng/mL) but not with genotype (carrier, 8.01±0.59 ng/mL; non-carrier, 7.17±0.64 ng/mL). Embryo number was greater in carriers (Exp1, 1.64±0.81; Exp2, 1.45±0.09) vs non-carriers (1.00±0.00, both Experiments). Single, twin, and triplet pregnancies occurred in carriers in Experiment 1 whereas multiples in Experiment 2 were limited to twin pregnancies. Genotype effects on pregnancy rate were not significant (p >0.10) in either experiment. Results suggest that follicular ablation to create bilateral twin pregnancies in Trio carriers is feasible but requires induction of accessory CL to offset the negative effects of follicular aspiration on subsequent P4 concentration and associated fertility outcomes.

Multiple birth in dairy cattle is a detrimental trait both economically for producers and for animal health. Genetics of twinning is complex and has led to several quantitative trait loci regions being associated with increased twinning. To identify variants associated with this trait, calving records from 2 time periods were used to estimate daughter averages for twinning for Holstein bulls. Multiple analyses were conducted and compared including GWAS, genomic prediction, and gene set enrichment analysis for pathway detection. Although pathway analysis did not yield many congruent pathways of interest between data sets, it did indicate two of interest. Both pathways have ties to the strong candidate region on BTA11 from the genomewide association analysis across data sets. This region does not overlap with previously identified quantitative trait loci regions for twinning or ovulation rate in cattle. The strongest associated SNPs were upstream from 2 candidate genes LHCGR and FSHR, which are involved in folliculogenesis. Genomic prediction showed a moderate correlation accuracy (0.43) when predicting genomic breeding values for bulls with estimates from calving records from 2010 to 2016. Future analysis of the region on BTA11 and the relation of the candidate genes could improve this accuracy.