Serum Vitamin D Is Associated with Antioxidant Potential in Peri-Parturient Cows

Dairy cows experience increased oxidative stress during periods of transition such as at the cessation of lactation and around the periparturient period, thus increasing disease risk. Despite routine supplementation of transition cow diets with certain vitamins in an attempt to mitigate oxidative stress, there is no currently available data directly linking vitamin supplementation with antioxidant potential (AOP) in transition cows. The objective of this study was to determine the association between serum vitamins and biomarkers of oxidative stress in healthy cows. Blood samples were collected from 240 cows at dry off (DO), close up (CU), and 2–10 days post-calving (DIM2-10). Blood samples were analyzed for vitamins (A, D, E), β-carotene, reactive oxygen species (ROS), and AOP. Spearman correlations and mixed linear regression models were used to assess associations between vitamins and measures of oxidant status. Vitamin D concentrations were positively associated with AOP at the CU and DIM2-10. Based on the positive association with AOP, additional in-vitro studies were conducted that showed vitamin D mitigated barrier integrity loss in endothelial cells during oxidative stress. These results indicate for the first time that vitamin D may have a role in promoting antioxidant potential in transition dairy cows

A randomized control trial to test the effect of pegbovigrastim treatment at dry-off on plasma and milk oxylipid profiles during early mammary gland involution and the postparturient period

ABSTRACT: The early period of mammary gland involution is a critical juncture in the lactation cycle that can have significant effects on milk production and mammary gland health. Pegbovigrastim (PEG) administered 1 wk prior and on the day of parturition can enhance immune function and reduce the incidence of mastitis in the early postpartum period. Oxylipids are potent metabolites of polyunsaturated fatty acids (PUFA) and are important mediators of inflammation. The objective of this study was to evaluate effects of PEG given 1 wk before and at the day of dry-off (D0) on concentrations of oxylipids in plasma and milk from 7 d before D0 to 14 d after, as well as the effects during the first 14 d of the subsequent lactation. We hypothesized that both pro- and anti-inflammatory oxylipids would vary based on initiation of mammary gland involution and that pegbovigrastim would affect oxylipid concentrations, particularly those related to leukocytes. A complete randomized blocked design was used to enroll cows into either a PEG treatment group (n = 10) or control group (n = 10; CON). Blood samples were collected −7, −2, −1, 0, 1, 2, 4, 7, and 14 d relative to dry-off and 5, 10, and 14 d postcalving. Samples were analyzed for PUFA and oxylipids in milk and plasma by ultra-performance mass spectrometry and liquid chromatography tandem quadrupole mass spectrometry, respectively. Overall, 30 lipid mediators were measured in both milk and plasma. Repeated measures analyses revealed a significant interaction of treatment by time for milk 8-iso-keto-15-prostaglandin E2, prostaglandin F2α, plasma 8,12-iso-prostaglandin Fα-VI, 11-hydroxyeicosatetraenoic acid, and 12-hydroxyheptadecatienoic acid. The majority of milk PUFA and oxylipids differed significantly during early mammary gland involution and into the early postpartum period. This study demonstrated changes in oxylipids in milk secretions and plasma during early involution, and further investigation may illuminate multiple complex processes and reveal targets for optimization of mammary gland involution

Activity of sEH and Oxidant Status during Systemic Bovine Coliform Mastitis.

Bovine coliform mastitis presents treatment challenges because of systemic inflammation and oxidative stress. Soluble epoxide hydrolase (sEH) is a promising therapeutic target in conditions characterized by inflammation and oxidative stress but has not been evaluated in cattle. We compared sEH activity and oxidant status in healthy Holstein dairy cows to those with systemic coliform mastitis (n = 5/group) using complementary approaches. First, the activity of sEH on [3H]-trans-diphenyl-propene oxide (tDPPO) was assessed ex vivo using tissue homogenates (mammary, liver, and kidney). Second, the concentrations of sEH substrates and metabolites in plasma, milk, and urine were determined as an index of in vivo sEH activity. Oxidant status was assessed in serum and milk. Data were analyzed by non-parametric methods. Metabolism of tDPPO was greater in mammary tissues from cows with coliform mastitis compared to controls. In contrast, ratios of sEH substrates and metabolites predicted lower sEH activity in cows with coliform mastitis than controls. Milk oxidant status showed greater prooxidant levels in coliform mastitis cows. Cows with coliform mastitis exhibit increased sEH activity in mammary tissue; at the same time, milk oxidant status is increased. Future studies should characterize sEH activity and oxidant status patterns and explore therapies targeting sEH during coliform mastitis

Differences in the Oxylipid Profiles of Bovine Milk and Plasma at Different Stages of Lactation

Mastitis is caused by a bacterial infection of the mammary gland, which reduces both milk quality and quantity produced for human consumption. The incidence and severity of bovine mastitis are greatest during the periparturient period that results from dysfunctional inflammatory responses and causes damage to milk synthesizing tissues. Oxylipids are potent fatty acid-derived mediators that control the onset and resolution of the inflammatory response. The purpose of this study was to investigate how oxylipid profiles change in bovine milk at different stages of the lactation cycle. Results showed significantly lower concentrations of both milk polyunsaturated fatty acid content and total oxylipid biosynthesis during early lactation when compared to mid- or late-lactation. The only oxylipid that was higher during early lactation was 20-hydroxyeicosatetraenoic acid (HETE), which is often associated with inflammatory-based diseases. Milk oxylipid profiles during the different stages of lactation differed from plasma profiles. As such, plasma fatty acid and oxylipid concentrations are not a proxy for local changes in the mammary gland during the lactation cycle

Linoleic acid derived oxylipid contents in plasma and adipose tissues from dairy cows are modified during the periparturient period.

<p>Adipose and plasma samples were collected at -27±7 (G1) and -10±5 d (G2) prepartum and at 8±3 d postpartum (PP). Cows were grouped by lipolysis intensity (LI) based on FFA concentrations at PP in low (LL = FFA<1.0 mEq/L) and high (HL, FFA≥1.0 mEq/L) lipolysis rate.</p

Selection of Potent Inhibitors of Soluble Epoxide Hydrolase for Usage in Veterinary Medicine.

The veterinary pharmacopeia available to treat pain and inflammation is limited in number, target of action and efficacy. Inhibitors of soluble epoxide hydrolase (sEH) are a new class of anti-inflammatory, pro-resolving and analgesic drugs being tested in humans that have demonstrated efficacy in laboratory animals. They block the hydrolysis, and thus, increase endogenous concentrations of analgesic and anti-inflammatory signaling molecules called epoxy-fatty acids. Here, we screened a library of 2,300 inhibitors of the sEH human against partially purified feline, canine and equine hepatic sEH to identify inhibitors that are broadly potent among species. Six very potent sEH inhibitors (IC50 &lt; 1 nM for each enzyme tested) were identified. Their microsomal stability was then measured in hepatic extracts from cat, dog and horse, as well as their solubility in solvents suitable for the formulation of drugs. The trans-4-{4-[3-(4-trifluoromethoxy-phenyl)-ureido]-cyclohexyloxy}-benzoic acid (t-TUCB, 1,728) appears to be the best compromise between stability and potency across species. Thus, it was selected for further testing in veterinary clinical trials of pain and inflammation in animals

Dynamics of circulating and adipose tissue fatty acids in periparturient dairy cows at -27±7 (G1) and -10±5 d (G2) prepartum and at 8±3 d postpartum (PP).

<p>Cows were grouped by lipolysis intensity (LI) based on FFA concentrations at PP in low (LL = FFA<1.0 mEq/L) and high (HL, FFA≥1.0 mEq/L) lipolysis rate.</p

Periparturient lipolysis and oxylipid biosynthesis in bovine adipose tissues

<div><p>The periparturient period of dairy cows is characterized by intense lipolysis in adipose tissues (AT), which induces the release of free fatty acids (FFA) into circulation. Among FFA, polyunsaturated fatty acids are susceptible to oxidation and can modulate inflammatory responses during lipolysis within AT. Linoleic and arachidonic acid oxidized products (oxylipids) such as hydroxy-octadecadienoic acids (HODE) and hydroxy-eicosatetraenoic acids (HETE), were recently identified as products of lipolysis that could modulate AT inflammation during lipolysis. However, the effect of lipolysis intensity during the transition from gestation to lactation on fatty acid substrate availability and subsequent AT oxylipid biosynthesis is currently unknown. We hypothesized that in periparturient dairy cows, alterations in AT and plasma fatty acids and oxylipid profiles coincide with changes in lipolysis intensity and stage of lactation. Blood and subcutaneous AT samples were collected from periparturient cows at -27±7 (G1) and -10±5 (G2) d prepartum and at 8±3 d postpartum (PP). Targeted lipidomic analysis was performed on plasma and AT using HPLC-MS/MS. We report that FFA concentrations increased as parturition approached and were highest at PP. Cows exhibiting high lipolysis rate at PP (FFA>1.0 mEq/L) had higher body condition scores at G1 compared to cows with low lipolysis rate (FFA<1.0 mEq/L). Concentrations of plasma linoleic and arachidonic acids were increased at PP. In AT, 13-HODE, and 5-, 11- and 15-HETE were increased at PP compared to G1 and G2. Concentrations of beta hydroxybutyrate were positively correlated with those of 13-HODE and 15-HETE in AT. Plasma concentrations of 5- and 20-HETE were increased at PP. These data demonstrate that prepartum adiposity predisposes cows to intense lipolysis post-partum and may exacerbate AT inflammation because of increased production of pro-inflammatory oxylipids including 5- and 15-HETE and 13-HODE. These results support a role for certain linoleic and arachidonic acid-derived oxylipids as positive and negative modulators of AT inflammation during periparturient lipolysis.</p></div

Partial correlation coefficients for circulating concentrations of free fatty acids (FFA) and β hydoxybutyrate (BHB) and contents of specific plasma FA and adipose tissue oxylipids, and gene expression of oxylipid enzymes and inflammatory markers in periparturient cows.

<p>Partial correlation coefficients for circulating concentrations of free fatty acids (FFA) and β hydoxybutyrate (BHB) and contents of specific plasma FA and adipose tissue oxylipids, and gene expression of oxylipid enzymes and inflammatory markers in periparturient cows.</p

Arachidonic acid-derived oxylipid contents in plasma and adipose tissues from dairy cows are modified during the periparturient period.

<p>Adipose and plasma samples were collected at -27±7 (G1) and -10±5 d (G2) prepartum and at 8±3 d postpartum (PP). Cows were grouped by lipolysis intensity (LI) based on FFA concentrations at PP in low (LL = FFA<1.0 mEq/L) and high (HL, FFA≥1.0 mEq/L) lipolysis rate.</p