Period In Dairy Cows Research Articles

Adipose Tissue Oxylipin Profile Changes with Subclinical Ketosis and Depot in Postpartum Dairy Cows

Lipolysis of adipose tissue is a natural occurrence during the periparturient period in dairy cows. However, when lipolysis rates exceed the capacity of other tissues to utilize nonesterified fatty acids (NEFA), it may lead to the development of ketosis and other diseases. Additionally, polyunsaturated NEFA (PUFA) can become oxidized into oxylipins, which modulate inflammation and metabolism. The objective of this work was to identify depot-specific differences on adipose tissue oxylipin profile in dairy cows with and without subclinical ketosis and assess the effects of oxylipins on adipocyte function in vitro. Subcutaneous adipose tissue from the flank (SAT) and visceral adipose tissue from the omentum (VAT) were collected through laparotomy from multiparous dairy cows (5-14 d in milk) and grouped according to blood β-hydroxybutyrate (BHB) into non-ketotic (NK; n = 5; BHB ≤ 0.8 mmol/L) and subclinical ketotic (SCK; n = 5; BHB 1.4 and ≤ 2.6 mmol/L). A targeted lipidome capable of detecting a 154 oxylipins was performed in paired SAT and VAT samples from all animals. Data were analyzed using the PROC GLIMMIX procedure in SAS (v9.4, SAS Institute Inc., Cary, NC, USA) for the effect of depot (SAT, VAT), ketosis status (SCK, NK), and their interaction (depot × ketosis status) on oxylipin abundance. The oxylipins thromboxane-B2 (TXB2), prostaglandin-A2 (PGA2), and 5-hydroxeicostretanoic acid (5-HETE) were selected from lipidomic data based on effects of ketosis status and depot-specificity to further investigate their effects on SAT and VAT adipocyte function. Lipidomic data revealed 50 oxylipins across both adipose tissue depots. SCK was associated with a decreased abundance of TXB2 and tended to associate with an increase in prostaglandinA2 (PGA2) and prostaglandinE1 (PGE1). Additionally, PGE1, 15-keto-prostaglandin-E2 (15-Keto-PGE2), 13,14-dihydro-15-keto-prostaglandin-E2 (13-14dh-15k-PGE2), 5-HETE, and 15-HETE were increased in SAT. While VAT had a greater abundance of 9,10-dihydroxy-12Z-octadecenoic acid (9,10-DiHOME), 12,13-dihydroxy-9Z-octadecenoic acid (12,13-DiHOME), 9-oxo-10E,12Z,15Z-octadecatrienoic acid (9-OxoOTrE) and 13S-hydroxy-9Z,11E,15Z-octadecatrienoic acid (13(s)-HOTrE). In vitro, AVG dose of 5-HETE on VAT cells tended to increase proliferation at d 7 compared with the control, HIGH dose of TXB2 tended to decrease lipid accumulation in SAT compared with control, and AVG dose of PGA2 on VAT cells tended to lower ROS compared with the control. Overall, postpartum dairy cows have depot-specific adipose tissue lipidomic profiles which are associated with changes in ketosis status.

Update on ketosis in dairy cattle with major emphasis on subclinical ketosis and abdominal adiposity.

The metabolic changes that occur during the early post-partum period in dairy cows can indeed lead to an imbalance in energy utilization, resulting in the production of excessive ketone bodies. This can have detrimental effects on the cow's health and milk production, leading to economic losses for dairy producers. The release of non-esterified fatty acids into the blood due to increased lipolysis is a key factor in the development of ketosis. Abdominal adiposity is a key factor on these outcomes in modern dairy cows. The redirection of energy and glucose towards lactose synthesis and milk yield leaves a deficit of gluconeogenic precursors, leading to the conversion of acetyl-CoA into ketone bodies instead of entering the Krebs cycle. These ketone bodies, including acetone, acetoacetate and β-hydroxybutyrate, accumulate in the blood and can be detected in various bodily fluids, such as urine, blood and milk, allowing for diagnostic testing. Prevention is indeed crucial in managing ketosis in dairy cattle. Supplementation of propylene glycol in the diet or the use of monensin, either in the diet or in the form of a slow-release bolus, can help prevent the occurrence of ketosis. However, avoiding high body condition (subcutaneous fat) and excessive abdominal adiposity during the dry period and parturition plus an adequate cow comfort are fundamental tasks to avoid ketosis and related disorders. These interventions aim to provide additional energy sources or enhance the cow's ability to utilize energy efficiently, thus reducing the reliance on excessive lipolysis and ketone body production.

Transition Period in Dairy Cows: Monitoring, Challenges, and Future Perspectives

Transition Period in Dairy Cows: Monitoring, Challenges, and Future Perspectives

Effect of Postbiotic Supplementation on Nutrient Digestibility and Milk Yield during the Transition Period in Dairy Cows.

The metabolites secreted by probiotics or released after their lysis are called postbiotics. They provide physiological benefits to the host, preventing the colonisation of pathogens by improving the intestinal environment for beneficial commensal bacteria, which reduces the incidence of digestive disorders and improves the immune system. The aim of this work was to evaluate the addition of postbiotics to dairy cow rations during the transition period on nutrient digestibility, composition, and milk yield. The effects of two postbiotics were evaluated in twelve Friesian cows from 30 days before calving to two months of lactation. The animals were randomly allocated to two treatment groups: control (CT) and supplemented with postbiotics (PC and PR). Feeding was ad libitum with 60/40 of forage/concentrate ratio on dry matter basis. Daily feed intake and milk production were recorded individually throughout the study. Two digestibility balances were performed, one before parturition and one after parturition. Colostrum was sampled at first milking and milk was sampled weekly. Data were analysed using a mixed mode in R software 4.4.1. The results suggest that postbiotic supplementation in late gestation and early lactation increase the voluntary intake of dry matter, especially in the PR treatment, with higher apparent total tract digestibility of dry matter, organic matter and neutral detergent fibre. Both treatments including postbiotics induced an increase in colostral immunoglobulin concentration. Milk production of cows receiving the PC treatment was the highest, with high fat and protein yields and a higher persistence of the production curve throughout the lactation.

Time profiles of energy balance in dairy cows in association with metabolic status, inflammatory status, and disease

The early lactation period in dairy cows is characterized by complex interactions among energy balance (EB), disease, and alterations in metabolic and inflammatory status. The objective of this study was to cluster cows based on EB time profiles in early lactation and investigate the association between EB clusters and inflammatory status, metabolic status, oxidative stress, and disease. Holstein-Friesian dairy cows (n = 153) were selected and monitored for disease treatments during wk 1 to 6 in lactation. Weekly EB was calculated based on energy intake and energy requirements for maintenance and milk yield in wk 1 to 6 in lactation. Weekly plasma samples were analyzed for metabolic variables in wk 1 to 6, and inflammatory and oxidative stress variables in wk 1, 2, and 4 in lactation. Liver activity index (LAI) was computed from plasma albumin, cholesterol, and retino-binding protein concentration. First, cows were clustered based on time profiles of EB, resulting in 4 clusters (SP: stable positive; MN: mild negative; IN: intermediate negative; SN: severe negative). Cows in the SN cluster had higher plasma nonesterified fatty acids and β-hydroxybutyrate concentrations, compared with cows in the SP cluster, with the MN and IN cluster being intermediate. Cows in the SN cluster had a higher milk yield, lower dry matter intake in wk 1, lower insulin concentration compared with cows in the SP cluster, and lower glucose and IGF-1 concentration compared with cows in the SP and MN clusters. Energy balance clusters were not related with plasma haptoglobin, cholesterol, albumin, paraoxonase, and liver activity index (LAI). Second, cows were grouped based on health status [IHP: cows with treatment for inflammatory health problem (endometritis, fever, clinical mastitis, vaginal discharge or retained placenta); OHP: cows with no IHP but treatment for other health problem (milk fever, cystic ovaries, claw, and leg problems, rumen and intestine problems or other diseases); NHP: cows with no treatments, in the first 6 weeks after calving]. Energy balance was not different among health status groups. The IHP cows had lower nonesterified fatty acids and greater insulin concentration in plasma compared with OHP. The IHP cows had lower plasma albumin concentration, lower LAI and higher haptoglobin concentration compared with OHP and NHP. Overall, EB time profiles were associated with the metabolic status of dairy cows in early lactation, but were only limitedly related with markers of inflammation and oxidative stress status. Inflammatory and metabolic status were related to disease events in early lactation and caused prolonged effects on liver metabolism.

Efficacy of internal and external teat sealants on cure and new infection risk in dry-off protocols for Holstein cows

Internal teat sealant products have been used alone or in combination with antibiotic dry cow treatment to prevent new IMI over the dry period in dairy cows. Conversely, knowledge about the efficacy of external teat sealants in the prevention of IMI is scarce. The objective of our study was to investigate the effect of 2 different teat sealants, one internal teat sealant and one external teat sealant, on the (1) risk of new IMI during the dry period, (2) risk of IMI cure, (3) first test day linear somatic cell score, (4) first test day milk yield, (5) incidence of farm-diagnosed clinical mastitis ≤ 30 DIM, and (6) incidence of culling ≤ 30 DIM. In a randomized clinical trial, Holstein cows (n = 1,500) from one commercial dairy were assigned to treatment and control groups. At dry off, cows in the treatment groups received an antibiotic dry cow treatment in combination with either an internal teat sealant (INT) or a single application of an external teat sealant (EXT). Control (CON) cows received the antibiotic dry cow treatment alone. Data on linear somatic cell score from the last DHI test day before dry off and the first test after calving, first test day milk yield, and the occurrence of farm-diagnosed clinical mastitis and culling ≤ 30 DIM were obtained from the farm management program. New IMI and cure of IMI during the dry period were calculated. Linear somatic cell score (mean ± SD) at first test day after calving differed among groups and was 3.2 ± 2.2 in CON, 2.8 ± 2.0 in INT, and 3.0 ± 2.1 in EXT groups. The risk of new IMI differed among groups and was 30.2% for CON cows, 18.2% for INT cows, and 22.6% for EXT cows. A Poisson regression analysis revealed that, compared with CON cows, the risks of new IMI were 40% lower for INT cows (risk ratio (RR) = 0.60, 95% CI = 0.41 to 0.88) and 25% lower for EXT cows (RR = 0.75, 95% CI = 0.52 to 1.07). By contrast, no meaningful differences were documented for cure risk, clinical mastitis risk during the first 30 DIM, or culling risk within the first 30 DIM. In summary, cows dried off with an INT in combination with antibiotic dry cow treatment had a lower linear somatic cell score at first test day after calving and reduced risk of new IMI than cows dried off with an EXT in addition to antibiotic dry cow treatment or cows dried off using antibiotic dry cow treatment alone. Further, we found supporting evidence that cows dried off with an EXT in addition to antibiotic dry cow treatment might have an advantage in reduced new IMI over cows dried off with an antibiotic dry cow treatment alone.

Periparturient blood T-lymphocyte PD-1 and CTLA-4 expression as potential predictors of new intramammary infections in dairy cows during early lactation (short communication)

BackgroundThe periparturient period in dairy cows is marked by immunosuppression which increases the likelihood of infectious disorders, particularly also mastitis. An in-depth understanding of peripartum leukocyte biology is vital for the implementation of highly successful post-partum disease prevention measures. Immune checkpoint molecules, such as programmed death 1 (PD-1) and cytotoxic T-lymphocyte antigen 4 (CTLA-4), are critical inhibitory receptors expressed on immune cells, particularly T cells, that drive immunosuppressive signaling pathways. However, the potential role of immune checkpoint molecules expression in T-cells on udder health has never been explored. Thus, the association between the occurrence of new postpartum intramammary infections (IMIs) and the expression of programmed cell death protein 1 (PD-1) and cytotoxic T-lymphocyte-associated antigen-4 (CTLA-4) on blood T-cells during the peripartum period was investigated.ResultsIn this study, the incidence of IMIs by any pathogen in early lactation was not associated with a higher expression of PD-1 and CTLA-4 in the periparturient period. However, the incidence of IMIs by major pathogens throughout the first month of lactation was significantly associated with higher expression of PD-1 at 14 days before calving (P = 0.03) and CTLA-4 at parturition (P = 0.03) by blood T-cells. Also, the expression of CTLA-4 at D0 (P = 0.012) by T-cells was associated with the occurrence of persistent IMIs during the first month of lactation.ConclusionsTo our knowledge, this is the first report to investigate the expression of PD-1 and CTLA-4 by blood T-lymphocytes during the periparturient period in dairy cows and to explore their relationship with the incidence of new IMIs in the postpartum period. Thus, a comprehensive understanding of leukocyte biology during peripartum would appear to be a prerequisite for the identification of resilient dairy cows or targets innovative (immunological) non-antibiotic approaches in the transition period.

The Effect of Intrauterine Administration of Zeolite on the Puerperal Period in Dairy Cows

Background: The development of new protocols has become a priority today in the context of the adverse effects of conventional treatments. Zeolite falls into this category due to its numerous applications reported in the specialized literature. In this study, the main expected objectives were to demonstrate that intrauterine administration of zeolite can contribute to improving the indicators underlying the development of pathologies during the postpartum period. Methods: To determine the dosage and potential toxic effects on the uterine mucosa, we conducted a study on rats, to which we administered different doses on the second day after delivery. Ultrasound measurements of the uterus, collection of bacteriological samples and uterine tissue were performed at 7, 14 and 21 days. The results were satisfactory in terms of tissue effects, with the 20% dosage being the most suitable. Following the completion of the preliminary study, we grouped the animals into two equal groups (n=32). Intrauterine administration of zeolite was performed on the second day after delivery and monitoring of uterine involution was carried out on days 7, 14 and 21 of the puerperium. At 14 days, we collected samples to determine the number of bacteria and uterine tissue. Result: The data obtained indicate that the group receiving intrauterine zeolite (n=16) experienced a decrease in the number of bacteria, significantly faster uterine involution on days 14 and 28, a more favourable uterine score and a significantly shorter service period. However, no significance was established regarding the number of doses of seminal material used.

Heart rate variability and behavioral alterations during prepartum period in dairy cows as predictors of calving: a preliminary study.

Parturition is crucial for dams, their calves, and cow managers. The prediction of calving time, which assists cow managers to decide on the relocation of cows to maternity pens and necessity of human supervision, is a pivotal aspect of livestock farming. However, existing methods of predicting calving time in dairy cows based on hormonal changes and clinical symptoms are time-consuming and yield unreliable predictions. Accordingly, we investigated whether heart rate variability (HRV) which is a non-invasive assessment of autonomic nervous system (ANS) activity and behavior during the prepartum period would be useful for predicting calving time in dairy cows. Eight pregnant cows were surveilled under electrocardiogram and video recordings for HRV and behavioral analyses, respectively. HRV parameters in time and frequency domains were evaluated. A 24-h time budget was calculated for each of six types of behavior (standing and lying with or without rumination, sleeping, and eating). Heart rate on calving day is considerably higher than those recorded on the days preceding calving. Low frequency power declined, whereas high frequency power escalated on the calving day compared to the period between 24 and 48 h before calving. The time budget for ruminating while lying decreased and that while standing increased markedly on the calving day compared to those allocated on the preceding days; nonetheless, the total time budget for ruminating did not differ during the prepartum period. We elucidated the ANS activity and behavioral profiles during prepartum period. Our results confirm that HRV parameters and behavior are useful for predicting calving time, and interestingly indicate that the time budget for ruminating while standing (or lying) may serve as a valuable predictor of calving. Collectively, our findings lay the foundation for future investigations to determine other potential predictors and formulate an algorithm for predicting calving time.

Haemato-biochemical changes associated with peri-parturient period in dairy cows

Haemato-biochemical changes associated with peri-parturient period in dairy cows

Dynamic fecal microenvironment properties enable predictions and understanding of peripartum blood oxidative status and nonesterified fatty acids in dairy cows

The transition period in dairy cows is a critical stage and peripartum oxidative status, negative energy balance (NEB), and inflammation are highly prevalent. Fecal microbial metabolism is closely associated with blood oxidative status and nonesterified fatty acids (NEFA) levels. Here, we investigated dynamic changes in total oxidative status markers and NEFA in blood, fecal microbiome, and metabolome of 30 dairy cows during transition (-21, -7, +7, +21 d relative to calving). Then the Bayesian network and 9 machine-learning algorithms were applied to dismantle their relationship. Our results show that the oxidative status indicator (OSI) of -21, -7, +7 d was higher than +21 d. The plasma concentration of NEFA peaked on +7 d. For fecal microenvironment, a decline in bacterial α diversity was observed at postpartum and in bacterial interactions at +7 d. Conversely, microbial metabolites involved in carbohydrate, lipid, and energy metabolism increased on +7 d. A correlation analysis revealed that 11 and 10 microbial metabolites contributed to OSI and NEFA variations, respectively (arc strength >0.5). The support vector machine (SVM) radial model showed the highest average predictive accuracy (100% and 88.9% in the test and external data sets) for OSI using 1 metabolite and 3 microbiota. The SVM radial model also showed the highest average diagnostic accuracy (100% and 91% in the test and external data sets) for NEFA with 2 metabolites and 3 microbiota. Our results reveal a relationship between variation in the fecal microenvironment and indicators of oxidative status, NEB, and inflammation, which provide a theoretical basis for the prevention and precise regulation of peripartum oxidative status and NEB.

Evaluating variations in metabolic profiles during the dry period related to the time of hyperketonemia onset in dairy cows.

Hyperketonemia (HYK) in early lactation can have a different impact on health and productivity depending on the timing of HYK onset. While specific metabolites measured during the dry period may serve as biomarkers of HYK, the correlations between metabolites represent a challenge for the use of metabolic profiles dataset, and little has been explored on HYK. This exploratory cohort study aimed a) to characterize the correlations among metabolites measured during the late dry period in dairy cows, and b) to identify biomarkers in the late dry period associated with the onset of HYK at the first (wk1) and second (wk2) week of lactation. Individual blood samples from 440 Holstein dairy cows were collected at 21 ± 3 days before expected parturition. From each sample, 36 different metabolites were measured in serum and plasma. Hyperketonemia was diagnosed in wk1 and wk2 of lactation based on the blood concentration of beta-hydroxybutyrate (BHB > 1.2 mmol/L). Principal component analysis (PCA) was performed to reduce metabolites to a smaller number of uncorrelated components. Multivariable logistic regression models were applied to assess the associations between principal components (PC) and HYK at wk1 only (HYK+ wk1), wk2 only (HYK+ wk2), or both weeks (HYK+ wk1-2). The incidence of HYK was 16.2% in the first week, 13.0% in the second week, and 21.2% within the first two weeks of lactation. The results of PCA highlighted 10 PCs from which two were associated with HYK+ wk1 as compared with cows without HYK during the first two weeks of lactation (non-HYK); the PC a2 led by bilirubin and non-esterified fatty acids (OR = 1.29; 95%CI: 1.02-1.68), and the PC a5 led by alkaline phosphatase (ALP) and gamma-glutamyl transferase (GGT) (OR = 2.77; 95%CI: 1.61-4.97). There was no evidence of an association between any PC and HYK+ wk2 (vs. non-HYK cows). Cows with elevated PC a5 (led by ALP and GGT) in the dry period were 3.18 times more likely to be HYK+ wk1 than HYK+ wk2 (OR: 3.18, 95%CI: 1.34-8.73; P = 0.013). Overall, the main hypothesis generated by our exploratory study suggests that cows with biomarkers of liver dysfunction (ALP, GGT, bilirubin) assessed by PCA at 3 weeks before calving are more likely to develop HYK during the first week of lactation compared to the second week. In addition, results suggest that cows with HYK in both of the first two weeks of lactation had an overall metabolic disbalance during the onset of the late dry period, which based on PCs, encompass biomarkers related to glucogenic and ketogenic metabolic pathways as well as liver dysfunction and fatty liver. Further research is needed to determine the underlying mechanisms associated with the different adaptations between cows that develop HYK during the first and second week of lactation.

Resveratrol reverses Palmitic Acid-induced cow neutrophils apoptosis through shifting glucose metabolism into lipid metabolism via Cav-1/ CPT 1-mediated FAO enhancement

Elevated plasma nonesterified fatty acids (NEFAs) affect neutrophils function and longevity during the periparturient period in dairy cows. Previous research has shown that resveratrol (RSV) may protect cell viability from NEFA-induced damage by regulating energy metabolism. However, it is unclear whether RSV has a protective effect on palmitic acid (PA)-treated neutrophils. The aim of this study was to investigate the molecular regulatory mechanism of the protective effect of RSV on neutrophils. The results showed that treatment with high concentrations of RSV (50 μM, 100 μM) maintained neutrophils activity by inhibiting neutrophils apoptosis (P < 0.05). Further analysis showed that high concentrations of RSV enhanced fatty acid oxidation (FAO) to produce ATP by promoting the expression of CAV1, ACSL-1 and CPT1 (P < 0. 05) while inhibiting glycolysis by suppressing PFK1 activity (P < 0. 05) and reducing glucose transport-related protein (GLUT1/GLUT4) expression by inhibiting glucose uptake (P < 0.05). These results suggest that RSV protects neutrophils from PA-induced apoptosis by regulating energy metabolism. Our results revealed that RSV protects neutrophils from PA-induced apoptosis by shifting glucose metabolism to lipid metabolism. This study tenders to a meaningful understanding of the effects of RSV on neutrophils function in periparturient cows suffering from negative energy balance (NEB).

Feeding Flaxseed and Lupins during the Transition Period in Dairy Cows: Effects on Production Performance, Fertility and Biochemical Blood Indices

Flaxseed and lupin seed were offered as an alternative dietary approach in dairy cows, through the partial substitution of soybean meal. Milk production and fertility traits were investigated. A total of 330 animals were allocated into two groups, treated (n = 176) and control (n = 154). From each group, 30 animals were selected for hematological and cytological studies. The experimental feeding period lasted for 81 days (25 days prepartum and 56 days postpartum). The control ration (group C) contained corn, barley, soybean meal, rapeseed cake, corn silage and lucerne hay; whereas, in the treatment group (group T), 50% of the soybean meal was replaced by an equal mixture of flaxseed and lupins. The two rations were formulated to be isonitrogenous and isoenergetic. Milk samples were analyzed for chemical composition, somatic cell count (SCC) content and total colony forming units (CFU). Blood samples were collected, and serum was analyzed for non-esterified fatty acids (NEFA), acute phase proteins (haptoglobin and serum amyloid) and lipid oxidation indices, namely thiobarbituric-acid-reactive substances (TBARS) and catalase activity. To assess polymorphonuclear neutrophils (PMN) numbers, endometrial samples from each cow were collected on days 21 and 42. No difference was recorded between groups in milk yield (p &gt; 0.05). In multiparous cows, NEFA (mMol/L) concentrations were significantly lower in group T than in group C on day 14 (p &gt; 0.009) and on day 42 (p = 0.05), while no difference was detected in the group of primiparous cows. At all time points, serum TBARS and catalase values were similar in both groups (p &gt; 0.05). Multiparous cows in group T expressed the first postpartum estrus and conceived earlier than cows in group C (p ≤ 0.05). Between days 21 to 42 postpartum, the PMN reduction rate was higher in group T animals (p ≤ 0.05). Acute phase protein levels were in general lower in group T animals, and at specific time points differed significantly from group C (p ≤ 0.05). It was concluded that the partial replacement of soybean meal by flaxseed and lupins had no negative effect on milk yield or milk composition, and improved cow fertility; which, along with the lower cost of flaxseed and lupins mixture, may increase milk production profitability.

Gene co-expression network and differential expression analyses of subcutaneous white adipose tissue reveal novel insights into the pathological mechanisms underlying ketosis in dairy cows

Ketosis is a common nutritional metabolic disease during the perinatal period in dairy cows. Although various risk factors have been identified, the molecular mechanism underlying ketosis remains elusive. In this study, subcutaneous white adipose tissue (sWAT) was biopsied for transcriptome sequencing on 10 Holstein cows with type II ketosis [blood β-hydroxybutyric acid (BHB) >1.4 mmol/L; Ket group] and another 10 cows without type II ketosis (BHB ≤1.4 mmol/L; Nket group) at d 10 after calving. Serum concentrations of nonesterified fatty acids (NEFA) and BHB, as indicators of excessive fat mobilization and circulating ketone bodies, respectively, were significantly higher in the Ket group than in the Nket group. Aspartate transaminase (AST) and total bilirubin (TBIL), as indicators of liver damage, were higher in the Ket group than in the Nket group. Weighted gene co-expression network analysis (WGCNA) of the sWAT transcriptome revealed modules significantly correlated with serum BHB, NEFA, AST, TBIL, and total cholesterol. The genes in these modules were enriched in the regulation of the lipid biosynthesis process. Neurotrophic tyrosine kinase receptor type 2 (NTRK2) was identified as the key hub gene by intramodular connectivity, gene significance, and module membership. Quantitative reverse transcription PCR analyses for these samples, as well as a set of independent samples, validated the downregulation of NTRK2 expression in the sWAT of dairy cows with type II ketosis. NTRK2 encodes tyrosine protein kinase receptor B (TrkB), which is a high-affinity receptor for brain-derived neurotrophic factor, suggesting that abnormal lipid mobilization in cows with type II ketosis might be associated with impaired central nervous system regulation of adipose tissue metabolism, providing a novel insight into the pathogenesis underlying type II ketosis in dairy cows.

AdipoRon alleviates fatty acid–induced lipid accumulation and mitochondrial dysfunction in bovine hepatocytes by promoting autophagy

During the transition period in dairy cows, high circulating concentrations of nonesterified fatty acids (NEFA) increase hepatic lipid deposits and are considered a major pathological factor for liver damage. We investigated whether AdipoRon, a synthetic small-molecule agonist of adiponectin receptors 1 and 2 shown to prevent liver lipid accumulation in nonruminants, could alleviate NEFA-induced lipid accumulation and mitochondrial dysfunction. Bovine hepatocytes were isolated from 5 healthy Holstein female newborn calves (1 d of age, 30-40 kg, fasting), and independently isolated hepatocytes from at least 3 different calves were used for each subsequent experiment. The composition and concentration of NEFA used in this study were selected according to hematological criteria of dairy cows with fatty liver or ketosis. First, hepatocytes were cultured with various concentrations of NEFA (0, 0.6, 1.2, or 2.4 mM) for 12 h. In a second experiment, hepatocytes were treated with AdipoRon at different concentrations (0, 5, 25, or 50 μM for 12 h) and times (25 μM for 0, 6, 12, or 24 h) with or without NEFA (1.2 mM) treatment. In the last experiment, hepatocytes were treated with AdipoRon (25 μM), NEFA (1.2 mM), or both for 12 h after treatment with or without the autophagy inhibitor chloroquine. Hepatocytes treated with NEFA had increased protein abundance of sterol regulatory element-binding protein 1c (SREBP-1c) and mRNA abundance of acetyl-CoA carboxylase 1 (ACACA), and decreased protein abundance of peroxisome proliferator-activated receptor α (PPARA), proliferator-activated receptor gamma coactivator-1 α (PGC-1α), mitofusin 2 (MFN2), cytochrome c oxidase subunit IV (COX IV), and mRNA abundance of carnitine palmitoyltransferase 1A (CPT1A), along with lower ATP concentrations. AdipoRon treatment reversed these effects, suggesting this compound had a positive effect on lipid metabolism and mitochondrial dysfunction during the NEFA challenge. In addition, upregulated expression of microtubule-associated protein 1 light chain 3-II (LC3-II, encoded by MAP1LC3) and downregulated expression of sequestosome-1 (SQSTM1, also called p62) indicated that AdipoRon enhanced autophagic activity in hepatocytes. The fact that chloroquine impeded the beneficial effects of AdipoRon on lipid accumulation and mitochondrial dysfunction suggested a direct role for autophagy during NEFA challenge. Our results suggest that autophagy is an important cellular mechanism to prevent NEFA-induced lipid accumulation and mitochondrial dysfunction in bovine hepatocytes, which is consistent with other studies. Overall, AdipoRon may represent a promising therapeutic agent to maintain hepatic lipid homeostasis and mitochondrial function in dairy cows during the transition period.

Alterations in skeletal muscle abundance of protein turnover, stress, and antioxidant proteins during the periparturient period in dairy cows fed ethyl-cellulose rumen-protected methionine

Skeletal muscle turnover helps support the physiological needs of dairy cows during the transition into lactation. We evaluated effects of feeding ethyl-cellulose rumen-protected methionine (RPM) during the periparturient period on abundance of proteins associated with transport AA and glucose, protein turnover, metabolism, and antioxidant pathways in skeletal muscle. Sixty multiparous Holstein cows were used in a block design and assigned to a control or RPM diet from -28 to 60 d in milk. The RPM was fed at a rate of 0.09% or 0.10% of dry matter intake (DMI) during the prepartal and postpartal periods to achieve a target Lys:Met ratio in the metabolizable protein of ∼2.8:1. Muscle biopsies from the hind leg of 10 clinically healthy cows per diet collected at -21, 1, and 21 d relative to calving were used for western blotting of 38 target proteins. Statistical analysis was performed using the PROC MIXED statement of SAS version 9.4 (SAS Institute Inc.) with cow as random effect, whereas diet, time, and diet × time were the fixed effects. Diet × time tended to affect prepartum DMI, with RPM cows consuming 15.2 kg/d and controls 14.6 kg/d. However, diet had no effect on postpartum DMI (17.2 and 17.1 ± 0.4 kg/d for control and RPM, respectively). Milk yield during the first 30 d in milk was also not affected by diet (38.1 and 37.5 ± 1.9 kg/d for control and RPM, respectively). Diet or time did not affect the abundance of several AA transporters or the insulin-induced glucose transporter (SLC2A4). Among evaluated proteins, feeding RPM led to lower overall abundance of proteins associated with protein synthesis (phosphorylated EEF2, phosphorylated RPS6KB1), mTOR activation (RRAGA), proteasome degradation (UBA1), cellular stress responses (HSP70, phosphorylated MAPK3, phosphorylated EIF2A, ERK1/2), antioxidant response (GPX3), and de novo synthesis of phospholipids (PEMT). Regardless of diet, there was an increase in the abundance of the active form of the master regulator of protein synthesis phosphorylated MTOR and the growth-factor-induced serine/threonine kinase phosphorylated AKT1 and PIK3C3, whereas the abundance of a negative regulator of translation (phosphorylated EEF2K) decreased over time. Compared with d 1 after calving and regardless of diet, the abundance of proteins associated with endoplasmic reticulum stress (XBP1 spliced), cell growth and survival (phosphorylated MAPK3), inflammation (transcription factor p65), antioxidant responses (KEAP1), and circadian regulation (CLOCK, PER2) of oxidative metabolism was upregulated at d 21 relative to parturition. These responses coupled with the upregulation of transporters for Lys, Arg, and His (SLC7A1) and glutamate/aspartate (SLC1A3) over time were suggestive of dynamic adaptations in cellular functions. Overall, management approaches that could take advantage of this physiological plasticity may help cows make a smoother transition into lactation.

Effect of Anti-Mullerian Hormone, Metabolic Profile and Mineral Levels at Transition Period On The Calving – Conception Interval in Cows

The transition period in dairy cows is generally accepted as the period covering 3 weeks before calving and 3 weeks after calving. The aim of the study was to compare the profiles of β-hydroxybutyrate, calcium, magnesium, phosphorus, total cholesterol, total protein, triacylglycerol, free glycerol, serum lipid and serum protein with Anti-Mullerian Hormone (AMH) in dairy cows in the transition period, and to determine whether AMH to examine whether it can be used as a marker in the next insemination period. The cows whose blood samples were taken were followed up and it was determined that they became pregnant at the insemination and the study was terminated. According to the results obtained; It was determined that BHB and free glycerol, which are important markers of negative energy balance (NEB), have an effect on AMH concentration. However, it was determined that the concentration of magnesium and the ratio of cholesterol ester in serum total fat did not change much during the transition period. Our results suggest that AMH is a good biomarker of decreased follicular activity due to NEB in the transition period and that AMH can be used for herd weeding in reinsemination.

The effect of co-supplementation of fish oil and vitamin E on reproductive performance and metabolic profile during the transitional period in dairy cows

The objective of the present study was to determine the effect of oral supplementation of dairy cows with a combination of fish oil and vitamin E (Vit E) from approximately 7 days (d) before the expected calving date up to 21 days postpartum, on their reproductive performance and metabolic profile. Eighty Holstein multiparous cows were randomly assigned into one of two ration groups during the transitional period. The treatment group cows (n=40) received a transitional diet (pre-and post-partum based on the NRC 2001) supplemented with fish oil (FO, 100 g/ once per day +Vit E (8000IU/kg/d); and the control cows (n=40) received the same concentrate without FO. Blood samples were collected at 1 week before the expected calving date, and 1, 2 and 3 weeks postpartum. After a voluntary waiting period, all cows received timed artificial insemination (TAI) at 76-81 days in milk (DIM) following the PresynchOvSynch protocol. Pregnancy diagnosis was performed 30-40 days after TAI using a transrectal ultrasonography. Our results showed that the cows fed the FO+Vit E diet had a statistically higher first service pregnancy rate (46.3 % vs. 39.6%, P&lt;0.001) at 30-40 days after TAI than those fed the control diet. Also, cows that consumed the FO+Vit E treatment diet had lower late embryo loss at 40-70 days post insemination than the control. Plasma concentrations of triglycerides (d7, 14, and 21), cholesterol (d7 and 14), glucose (d14), insulin (d-7) progesterone (d14 and 21) in the treatment group were higher (P&lt;0.05) compared to the cows fed control diets. Non-esterified fatty acid (NEFA) levels were not significantly affected by the dietary treatments pre-partum, while β –hydroxybutirate acids (BHBA) levels on day 21 (P&lt;0.05) were higher in the control group. The results indicated that supplementation of the diet with fatty acid and high doses of vitamin E could improve reproductive performance in dairy cattle.

Is adenosine deaminase (ADA) activity in saliva and serum a more accurate disease detection tool than traditional redox balance parameters in early-lactating dairy cows?

Enzyme adenosine deaminase (ADA) is a marker of inflammation in domestic animals, but it is unclear whether it is a reliable marker of oxidative stress, especially in the transition period in dairy cows. This study aims to assess if ADA and redox status measurements in saliva provide the same utility to detect disease condition as that obtained from serum. Sixty-eight multiparous Holstein cows, between 2 and 3 weeks postpartum were selected. Five study groups were established: control (healthy), and cows with ketosis, mastitis, laminitis, and metritis. The parameters measured were ADA activity, total oxidants (TOS), antioxidants (TAC), and OSi ratio.Regarding redox status, no significant differences arise in both saliva and serum being the correlations negative and not significant. In saliva, ADA activity in healthy cows differs from those with pathological processes, having the lowest activities. In serum, ADA activity is similar in the healthy and ketosis cows, showing the lowest activities meanwhile animals with mastitis, laminitis, or metritis have significantly higher activities. In conclusion, the measurement of ADA activities and redox status in saliva does not give consistent results, being preferable to measure them in serum during the transition period.