In dairy cows, periparturient disease states, such as metritis, mastitis, and laminitis, are leading to increasingly significant economic losses for the dairy industry. Treatments for these pathologies are often expensive, ineffective, or not cost-efficient, leading to production losses, high veterinary bills, or early culling of the cows. Early diagnosis or detection of these conditions before they manifest themselves could lower their incidence, level of morbidity, and the associated economic losses. In an effort to identify predictive biomarkers for postpartum or periparturient disease states in dairy cows, we undertook a cross-sectional and longitudinal metabolomics study to look at plasma metabolite levels of dairy cows during the transition period, before and after becoming ill with postpartum diseases. Specifically we employed a targeted quantitative metabolomics approach that uses direct flow injection mass spectrometry to track the metabolite changes in 120 different plasma metabolites. Blood plasma samples were collected from 12 dairy cows at 4 time points during the transition period (-4 and -1 wk before and 1 and 4 wk after parturition). Out of the 12 cows studied, 6 developed multiple periparturient disorders in the postcalving period, whereas the other 6 remained healthy during the entire experimental period. Multivariate data analysis (principal component analysis and partial least squares discriminant analysis) revealed a clear separation between healthy controls and diseased cows at all 4 time points. This analysis allowed us to identify several metabolites most responsible for separating the 2 groups, especially before parturition and the start of any postpartum disease. Three metabolites, carnitine, propionyl carnitine, and lysophosphatidylcholine acyl C14:0, were significantly elevated in diseased cows as compared with healthy controls as early as 4 wk before parturition, whereas 2 metabolites, phosphatidylcholine acyl-alkyl C42:4 and phosphatidylcholine diacyl C42:6, could be used to discriminate healthy controls from diseased cows 1 wk before parturition. A 3-metabolite plasma biomarker profile was developed that could predict which cows would develop periparturient diseases, up to 4 wk before clinical symptoms appearing, with a sensitivity of 87% and a specificity of 85%. This is the first report showing that periparturient diseases can be predicted in dairy cattle before their development using a multimetabolite biomarker model. Further research is warranted to validate these potential predictive biomarkers.

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Simple SummaryLameness is prevalent in dairy cows and early diagnosis and timely treatment of the disease can lower animal suffering, improve recovery rate, increase longevity, and minimize cow loss. However, there are no indications of disease until it appears clinically, and presently the only approach to deal with the sick cow is intensive treatment or culling. The results suggest that lameness affected serum concentrations of the several parameters related to innate immunity and carbohydrate metabolism that might be used to monitor health status of transition dairy cows in the near future.AbstractThe objectives of this study were to evaluate metabolic and innate immunity alterations in the blood of transition dairy cows before, during, and after diagnosis of lameness during periparturient period. Blood samples were collected from the coccygeal vain once per week before morning feeding from 100 multiparous Holstein dairy cows during −8, −4, disease diagnosis, and +4 weeks (wks) relative to parturition. Six healthy cows (CON) and six cows that showed clinical signs of lameness were selected for intensive serum analyses. Concentrations of interleukin-1 (IL-1), interleukin-6 (IL-6), tumor necrosis factor (TNF), haptoglobin (Hp), serum amyloid A (SAA), lipopolysaccharide binding protein (LBP), lactate, non-esterified fatty acids (NEFA), and β-hydroxybutyrate (BHBA) were measured in serum by ELISA or colorimetric methods. Health status, DMI, rectal temperature, milk yield, and milk composition also were monitored for each cow during the whole experimental period. Results showed that cows affected by lameness had greater concentrations of lactate, IL-6, and SAA in the serum vs. CON cows. Concentrations of TNF tended to be greater in cows with lameness compared with CON. In addition, there was a health status (Hs) by time (week) interaction for IL-1, TNF, and Hp in lameness cows vs. CON ones. Enhanced serum concentrations of lactate, IL-6, and SAA at −8 and −4 wks before parturition were different in cows with lameness as compared with those of the CON group. The disease was also associated with lowered overall milk production and DMI as well as milk fat and fat-to-protein ratio. In conclusion, cows affected postpartum by lameness had alterations in several serum variables related to innate immunity and carbohydrate metabolism that give insights into the etiopathogenesis of the disease and might serve to monitor health status of transition dairy cows in the near future.

BackgroundFailure to expel fetal membranes within 24 h of calving is a pathological condition defined as retained placenta (RP). The objective of this investigation was to evaluate whether there are alterations in several selected serum variables related to innate immunity and carbohydrate and lipid metabolism that precede occurrence of RP in transition Holstein dairy cows.MethodsOne hundred multiparous Holstein dairy cows were involved in the study. Blood samples were collected from the coccygeal vein during the −8 to +4 wks around parturition, once per week before the morning feeding. Six healthy control cows (CON) and 6 cows with RP were selected and serum samples at −8, −4, time of diagnosis of disease, and +4 wks relative to parturition were used for analyses. All samples were analyzed for lactate, non-esterified fatty acids (NEFA), β-hydroxybutyrate (BHBA), interleukin-1 (IL-1), interleukin-6 (IL-6), tumor necrosis factor (TNF), haptoglobin (Hp), and serum amyloid A (SAA).ResultsCows with RP had greater concentrations of serum lactate, IL-1, IL-6, TNF, and SAA in comparison with CON cows. Intriguingly, elevated concentrations of all five variables were observed at −8 and −4 wks before the occurrence of RP compared to healthy cows. Cows with RP also had lower DMI and milk production vs CON animals; however milk composition was not affected by RP.ConclusionsCows with RP showed an activated innate immunity 8 wks prior to diagnosis of disease. Overall results suggest that serum IL-1, IL- 6, and TNF, and lactate can be used as screening biomarkers to indicate cows that might have health issues during the transition period.

BackgroundThis study examined whether activation of innate immunity and alterations of carbohydrate and lipid metabolism precede development of subclinical mastitis (SCM).MethodsBlood samples were collected from the coccygeal vein from 100 Holstein dairy cows at -8, -4, disease diagnosis week, and +4 weeks postpartum. Six healthy cows (controls – CON) and six cows that showed clinical signs of SCM were selected for serum analyses. All serum samples were analyzed for acute phase proteins (APP) haptoglobin (Hp) and serum amyloid A (SAA); proinflammatory cytokines including interleukin 1 (IL-1), IL-6, and tumor necrosis factor (TNF) and serum lactate, BHBA, and NEFA concentration. Data of DMI, milk production, and milk composition were recorded and analyzed.ResultsThe results showed that cows with SCM had greater concentrations of SAA, TNF (P < 0.01), and lactate before expected day of parturition (P < 0.05) compared to CON cows. Cows with SCM showed greater concentrations of lactate starting at -8 weeks (P < 0.05) and TNF starting at -4 weeks prior to the expected day of parturition (P < 0.01). Interestingly, at -4 weeks, concentrations of IL-1 and Hp were lower in cows with SCM compared to healthy cows (P < 0.01) followed by an increase during the week of disease diagnosis (P < 0.05). Subclinical mastitis was associated with lower DMI, at -4 weeks before calving, milk production (P < 0.05) and increased somatic cell counts (SCC) (P < 0.01).ConclusionsResults of this study suggest that SCM is preceded by activated innate immunity and altered carbohydrate metabolism in transition dairy cows. Moreover the results support the idea that Hp, lactate, and SAA, at -8 weeks, and TNF and IL-1 at -4 weeks can be used as early indicators to screen cows during dry off for disease state.