ABSTRACT
The aim of this study was to identify with a high level of confidence metabolites previously identified as predictors of lameness and understand their biological relevance by carrying out pathway analyses. For the dairy cattle sector, lameness is a major challenge with a large impact on animal welfare and farm economics. Understanding metabolic alterations during the transition period associated with lameness before the appearance of clinical signs may allow its early detection and risk prevention. The annotation with high confidence of metabolite predictors of lameness and the understanding of interactions between metabolism and immunity are crucial for a better understanding of this condition. Using liquid chromatography-tandem mass spectrometry (LC-MS/MS) with authentic standards to increase confidence in the putative annotations of metabolites previously determined as predictive for lameness in transition dairy cows, it was possible to identify cresol, valproic acid, and gluconolactone as L1, L2, and L1, respectively which are the highest levels of confidence in identification. The metabolite set enrichment analysis of biological pathways in which predictors of lameness are involved identified six significant pathways (p < 0.05). In comparison, over-representation analysis and topology analysis identified two significant pathways (p < 0.05). Overall, our LC-MS/MS analysis proved to be adequate to confidently identify metabolites in urine samples previously found to be predictive of lameness, and understand their potential biological relevance, despite the challenges of metabolite identification and pathway analysis when performing untargeted metabolomics. This approach shows potential as a reliable method to identify biomarkers that can be used in the future to predict the risk of lameness before calving. Validation with a larger cohort is required to assess the generalization of these findings.
ABSTRACT
The transition period is a pivotal time in the production cycle of the dairy cow. It is estimated that between 30% and 50% of all cows experience metabolic or infectious disease during this time. One of the most common and economically consequential effects of disease during the transition period is a reduction in early lactation milk production. This has led to the utilisation of deviation from expected milk yield in early lactation as a proxy measure for transition health. However, to date, this analysis has been used exclusively for the retrospective assessment of transition cow health. Statistical models capable of predicting deviations from expected milk yield may allow producers to proactively manage animals predicted to suffer negative deviations in early lactation milk production. The objective of this retrospective cohort study was first, to explore the accuracy with which cow-level production and behaviour data collected on automatic milking systems (AMS) from 1-3 days in milk (DIM) can predict deviation from expected 30-day cumulative milk yield in multiparous cows. And second, to assess the accuracy with which predicted yield deviations can classify cows into groups which may facilitate improved transition management. Production, rumination, and physical activity data from 31 commercial AMS were accessed. A 3-step analytical procedure was then conducted. In Step 1, expected cumulative yield for 1-30 DIM for each individual cow-lactation was calculated using a mixed effect linear model. In Step 2, 30-Day Yield Deviation (YD) was calculated as the difference between observed and expected cumulative yield. Lactations were then assigned to one of three groups based on their YD, RED Group (= -15% YD), AMBER Group (-14% ̶ 0% YD), GREEN Group (>0% YD). In Step 3, yield, rumination, and physical activity data from days 1-3 in lactation were used to predict YD using machine learning models. Following external validation, YD was predicted across the test data set with a mean absolute error of 9%. Categorisation of animals suffering large negative deviations (RED group) was achieved with a specificity of 99%, sensitivity of 35%, and balanced accuracy of 67%. Our results suggest that milk yield, rumination and physical activity patterns expressed by dairy cows from 1-3 DIM have utility in the prediction of deviation from expected 30-day cumulative yield. However, these predictions currently lack the sensitivity required to classify cows reliably and completely into groups which may facilitate improved transition cow management.