Background Various blood metabolites are known to be useful indicators of health status in dairy cattle,but their routine assessment is time-consuming,expensive,and stressful for the cows at the herd level.Thus,we evaluated the effectiveness of combining in-line near infrared(NIR) milk spectra with on-farm(days in milk [DIM] and parity)and genetic markers for predicting blood metabolites in Holstein cattle.Data were obtained from 388 Holstein cows from a farm with an AfiLab system.NIR spectra,on-farm information,and single nucleotide polymorphisms(SNP)markers were blended to develop calibration equations for blood metabolites using the elastic net(ENet) approach,considering 3 mod els:(1) Model 1(M1) including only NIR information,(2) Model 2(M2) with both NIR and on-farm information,and(3) Model 3(M3)combining NIR,on-farm and genomic information.Dimension reduction was considered for M3 by preselecting SNP markers from genome-wide association study(GWAS) results.Results Results indicate that M2 improved the predictive ability by an average of 19% for energy-related metabolites(glucose,cholesterol,NEFA,B H B,urea,and c reatinin e),20% for liver functio n/hepatic damage,7% for inflammation/innate immunity.24% for oxidative stress metabolites,and 23% for minerals compared to M1,Meanwhile, M3 further enhanced the predictive ability by 34% for energy-related metabolites,32% for liver function/hepatic damage,22%for inflammation/innate immunity,42.1% for oxidative stress metabolites,and 41% for mineralse compared to M1.We found improved predictive ability of M3 using selected SNP markers from GWAS results using a threshold of >2.0by 5% for energy-related metabolites,9% for liver function/hepatic damage,8% for inflammation/innate immunity,22% for oxidative stress metabolites,and 9% for minerals.Slight redu ctions were observed fo r phosphorus(2%),ferricreducing antioxidant power(1%),and glucose(3%).Furthermore,it was found that prediction accuracies are influenced by using more restrictive thresholds(-log10(P-value)> 2.5 and 3.0),with a lower increase in the predictive ability.Conclusion Our results highlighted the potential of combining several sources of information,such as genetic markers,on-farm information,and in-line NIR infrared data improves the predictive ability of blood metabolites in dairy cattle,representing an effective strategy for large-scale in-line health monitoring in commercial herds.