Multiple β-defensin genes are upregulated by the vitamin D pathway in cattle

Extensive metabolic and physiologic changes occur during the peripartum, concurrent with a high incidence of infectious disease. Immune dysfunction is a likely contributor to the increased risk of disease at this time. Studies using high-yielding, total mixed ration-fed cows have indicated that neutrophil function is perturbed over the transition period; however, this reported dysfunction has yet to be investigated in moderate-yielding, grazing dairy cows. Therefore, we investigated changes in the expression of genes involved in neutrophil function. Blood was collected from cows at 5 time points over the transition period: precalving (-1wk; n=46), day of calving (d 0; n=46), and postcalving at wk 1 (n=46), wk 2 (n=45), and wk 4 (n=43). Neutrophils were isolated by differential centrifugation and gene expression was investigated. Quantitative reverse transcriptase PCR with custom-designed primer pairs and Roche Universal Probe Library (Roche, Basel, Switzerland) chemistry, combined with microfluidics integrated fluidic circuit chips (96.96 Dynamic Array, San Francisco, CA) were used to investigate the expression of 78 genes involved in neutrophil function and 18 endogenous control genes. Statistical significance between time points was determined using a repeated measures ANOVA. Genes that were differentially expressed over the transition period included those involved in neutrophil adhesion (SELL, ITGB2, and ITGBX), mediation of the immune response (TLR4, HLA-DRA, and CXCR2), maturation, cell cycle progression, apoptosis (MCL1, BCL2, FASLG, and RIPK1), and control of gene expression (PPARG, PPARD, and STAT3). We noted reduced gene expression of proinflammatory cytokines (IFNG, TNF, IL12, and CCL2) on the day of calving, whereas anti-inflammatory cytokine gene expression (IL10) was upregulated. Increased gene expression of antimicrobial peptides (BNBD4, DEFB10, and DEFB1) occurred on the day of calving. Collectively, transcription profiles are indicative of functional changes in neutrophils of grazing dairy cows over the transition period and align with studies in cows of conventional total mixed ration systems. This altered function may predispose cows to disease over the transition period and is likely to be a natural change in function due to parturition.

mentioning

confidence: 99%

Parturition in dairy cows temporarily alters the expression of genes in circulating neutrophils

Crookenden

Heiser

Murray

et al. 2016

“…Calcidiol has beneficial effects on innate host defenses of cattle (Nelson et al, 2010(Nelson et al, , 2012Merriman et al, 2015). Exposure of monocytes to pathogen- (Nelson et al, 2010).…”

Section: Discussionmentioning

confidence: 99%

“…Activation of toll-like receptors results in production of peptides with potent antimicrobial activity such as cathelicidin and β defensins, which can disrupt bacterial cell membrane and cause bacterial death (Liu et al, 2006). Recently, Merriman et al (2015) showed that culture of bovine monocytes treated with calcitriol showed increased expression of a cluster of antimicrobial peptides and the increase in antimicrobial peptide expression was dose-dependent. Moreover, the authors showed that intramammary infusion of calcitriol increased expression of β-defensin gene 7 in macrophages isolated from the mammary gland (Merriman et al, 2015), suggesting that calcitriol stimulates defensive mechanisms in bovine immune cells.…”

Section: Discussionmentioning

confidence: 99%

“…Recently, Merriman et al (2015) showed that culture of bovine monocytes treated with calcitriol showed increased expression of a cluster of antimicrobial peptides and the increase in antimicrobial peptide expression was dose-dependent. Moreover, the authors showed that intramammary infusion of calcitriol increased expression of β-defensin gene 7 in macrophages isolated from the mammary gland (Merriman et al, 2015), suggesting that calcitriol stimulates defensive mechanisms in bovine immune cells. Cows fed CA in our experiment had increased concentrations of 1,25-dihydroxyvitamin D 3 in the last 9 d of gestation compared with cows fed CH (Rodney et al, 2018).…”

Section: Discussionmentioning

confidence: 99%

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Effects of prepartum dietary cation-anion difference and source of vitamin D in dairy cows: Health and reproductive responses

Martínez

Rodney

Block³

et al. 2018

The objectives of the experiment were to evaluate the effects of feeding diets with distinct dietary cation-anion difference (DCAD) levels and supplemented with 2 sources of vitamin D during the prepartum transition period on postpartum health and reproduction in dairy cows. The hypotheses were that feeding acidogenic diets prepartum would reduce the risk of hypocalcemia and other diseases, and the benefits of a negative DCAD treatment on health would be potentiated by supplementing calcidiol compared with cholecalciferol. Cows at 252 d of gestation were blocked by parity (28 nulliparous and 52 parous cows) and milk yield within parous cows, and randomly assigned to 1 of 4 treatments arranged as a 2 × 2 factorial, with 2 levels of DCAD, positive (+130 mEq/kg) or negative (-130 mEq/kg), and 2 sources of vitamin D, cholecalciferol or calcidiol, fed at 3 mg for each 11 kg of diet dry matter. The resulting treatment combinations were positive DCAD with cholecalciferol (PCH), positive DCAD with calcidiol (PCA), negative DCAD with cholecalciferol (NCH), and negative DCAD with calcidiol (NCA), which were fed from 252 d of gestation to calving. After calving, cows were fed the same lactation diet supplemented with cholecalciferol at 0.70 mg for every 20 kg of dry matter. Blood was sampled 7 d before parturition, and at 2 and 7 d postpartum to evaluate cell counts and measures of neutrophil function. Postpartum clinical and subclinical diseases and reproductive responses were evaluated. Feeding a diet with negative DCAD eliminated clinical hypocalcemia (23.1 vs. 0%) and drastically reduced the incidence and daily risk of subclinical hypocalcemia, and these effects were observed in the first 48 to 72 h after calving. The diet with negative DCAD tended to improve the intensity of oxidative burst activity of neutrophils in all cows prepartum and increased the intensity of phagocytosis in parous cows prepartum and the proportion of neutrophils with killing activity in parous cows postpartum (58.5 vs. 67.6%). Feeding calcidiol improved the proportion of neutrophils with oxidative burst activity (60.0 vs. 68.7%), reduced the incidences of retained placenta (30.8 vs. 2.5%) and metritis (46.2 vs. 23.1%), and reduced the proportion of cows with multiple diseases in early lactation. Combining the negative DCAD diet with calcidiol reduced morbidity by at least 60% compared with any of the other treatments. Cows with morbidity had lower blood ionized Ca and serum total Ca concentrations than healthy cows. Treatments did not affect the daily risk of hyperketonemia in the first 30 d of lactation. Despite the changes in cow health, manipulating the prepartum DCAD did not influence reproduction, but feeding calcidiol tended to increase the rate of pregnancy by 55%, which reduced the median days open by 19. In conclusion, feeding prepartum cows with a diet containing a negative DCAD combined with 3 mg of calcidiol benefited health in early lactation.

“…Recent reports have documented positive effects of vitamin D in mitigation of mastitis (Lippolis et al, 2011) and metritis (Martinez et al, 2018a). In addition to the major role of vitamin D in Ca and P homeostasis, a direct immunomodulatory role of vitamin D is documented in innate immune cells (Nelson et al, 2010b;Merriman et al, 2015) and the mammary gland (Merriman et al, 2017) of cattle.…”

Section: Introductionmentioning

confidence: 99%

Feeding supplemental 25-hydroxyvitamin D3 increases serum mineral concentrations and alters mammary immunity of lactating dairy cows

Poindexter

Kweh

Zimpel

et al. 2020

Self Cite

Objectives were to determine the effects of feeding supplemental 25-hydroxyvitamin D 3 [25(OH)D 3 ] on concentrations of vitamin D metabolites and minerals in serum, mammary immune status, and responses to intramammary bacterial infection in dairy cows. Sixty multiparous, pregnant lactating Holstein cows with somatic cell count <200,000/mL were blocked by days in milk and milk yield and randomly assigned to receive a daily top-dressed dietary supplement containing 1 or 3 mg of vitamin D 3 (1mgD or 3mgD), or 1 or 3 mg 25(OH) D 3 (1mg25D or 3mg25D) for 28 d (n = 15/treatment). Cows were kept in a freestall barn and fed a total mixed ration in individual feeding gates. Individual dry matter intake (DMI) and milk yield were recorded daily, and milk and blood samples were collected at 0, 7, 14, and 21 d relative to the start of treatment. At 21 d, cows fed 1mgD and 3mg25D received an intramammary challenge with Streptococcus uberis. Cows were observed for severity of mastitis, and blood and milk samples were collected every 12 h to measure inflammation. The 1mg25D and 3mg25D cows had greater serum 25(OH)D 3 concentrations at 21 d compared with 1mgD and 3mgD cows (62 ± 7, 66 ± 8, 135 ± 15, and 232 ± 26 ng/mL for 1mgD, 3mgD, 1mg25D, and 3mg25D, respectively). The 3mg25D cows had greater concentrations of Ca and P in serum at 21 d compared with other treatments (Ca = 2.38, 2.4, 2.37, and 2.48 ± 0.02 mM, 1.87, 1.88, and 2.10 ± 0.08 mM for 1mgD, 3mgD, 1mg25D, and 3mg25D, respectively). Yields of milk and milk components, DMI, body weight, and concentrations of 1,25-dihydroxyvitamin D and Mg in serum did not differ among treatments. Abundance of mRNA transcripts for interleukin-1β (IL1B) and inducible ni-tric oxide synthase (iNOS) in milk somatic cells before S. uberis challenge were increased in cows fed 25(OH) D 3 compared with cows fed vitamin D 3. Furthermore, IL1B, iNOS, β-defensin 7, and β-defensin 10 in milk somatic cells increased as concentrations of 25(OH)D 3 increased in serum. Cows fed 3mg25D had less severe mastitis at 60 and 72 h after challenge with S. uberis compared with cows fed 1mgD. Concentrations of bacteria, somatic cells, and serum albumin in milk after challenge did not differ between treatments; however, an interaction between treatment and day was detected for lactate dehydrogenase in milk. Expression of adhesion protein CD11b on milk neutrophils after the S. uberis challenge was greater among 3mg25D cows compared with 1mgD cows. Transcripts of CYP24A1 and iNOS in milk somatic cells during mastitis also were greater in 3mg25D cows compared with 1mgD cows. Feeding 25(OH)D 3 increased serum 25(OH)D 3 more effectively than supplemental vitamin D 3 , resulting in increased serum mineral concentrations, increased expression of vitamin D-responsive genes, and altered immune responses to intramammary bacterial challenge.