Postpartum Negative Energy Balance Research Articles

Possibility to Estimate Same Day Energy Status of Dairy Cows during First Half of Lactation by Non-Invasive Markers with Emphasis to Milk Fatty Acids.

Postpartum negative energy balance (NEB) is detrimental to cows and decreases profitability in dairy farming. The two origins of milk fatty acids (FA), de novo synthesized in the mammary gland and plasma lipids initially originating from feed, rumen microbes and the animal's adipose tissue, make milk FA candidates as possible NEB biomarkers. The aim of this study was to assess the possibility to predict EB in cows in the first 150 days of lactation with BCS, milk traits and selected individual milk FA and the ratios of blood-derived and de novo synthesized FA. The daily EB of Estonian Holstein cows (N = 30) was calculated based on body weights and BCS values. Milk FA were analyzed with gas chromatography. The variance partitioning analysis revealed that milk production traits, BCS at calving, FA ratios and days in milk accounted for 67.1% of the EB variance. Random forest analysis indicated the highest impact of the ratios C18:1cis9/C12:0+C14:0, C18:1cis9+C18:0/C12:0+C14:0, C18:1cis9/C14:0, C18:1cis9+C18:0/C14:0, C18:1cis9/sum C5:0 to C14:0, C18:1cis9+C18:0/sum C5:0 to C14:0 or C18:1cis9/C15:0. FA and their ratios alone explained 63.6% of the EB variance, indicating the possibility to use milk FA and their ratios as sole predictors for the energy status in dairy cows.

The Effects of Postpartum Yak Metabolism on Reproductive System Recovery.

The goal of this study was to determine the metabolism of multiparous female yaks during the late perinatal period and identify its effects on reproductive recovery in order to explain the low reproduction rate of yaks. Eight multiparous female yaks were randomly selected as the sample, and serum was collected from the yaks every 7 days from the day of delivery until 28 days after the delivery (five time points). The presence of serum metabolic profiles and reproductive hormones was identified using ELISA. The key metabolites were identified using liquid chromatography-mass spectrometry, and a dynamic metabolic network representation was created using bioinformatics analysis. A total of 117 different metabolites were identified by calculating the fold change of the metabolite expression at each time point. The dynamic metabolic network was created to represent the activities of the key metabolites, metabolic indexes and reproductive hormones. The initial efficiency of the glucose metabolism in the late perinatal period was found to be low, but it increased during the final period. The initial efficiencies of the lipid and amino acid metabolisms were high but decreased during the final period. We inferred that there was a postpartum negative energy balance in female yaks and that the synthesis and secretion of estrogen were blocked due to an excessive fatty acid mobilization. As a result, the reproductive hormone synthesis and secretion were maintained at a low level in the late perinatal period, and this was the main reason for the delayed recovery of the reproductive function postpartum. However, the specific mechanism needs to be further verified.

Longitudinal liver proteome profiling in dairy cows during the transition from gestation to lactation: Investigating metabolic adaptations and their interactions with fatty acids supplementation via repeated measurements ANOVA-simultaneous component analysis

Repeated measurements analysis of variance – simultaneous component analysis (ASCA) has been developed to handle complex longitudinal omics datasets and combine novel information with existing data. Herein, we aimed at applying ASCA to 64 liver proteomes collected at 4-time points (day −21, +1, +28, and + 63 relative to parturition) from 16 Holstein cows treated from 9 wk. antepartum to 9 wk. postpartum (PP) with coconut oil (CTRL) or a mixture of essential fatty acids (EFA) and conjugated linoleic acid (CLA) (EFA + CLA). The ASCA modeled 116, 43, and 97 differentially abundant proteins (DAP) during the transition to lactation, between CTRL and EFA + CLA, and their interaction, respectively. Time-dependent DAP were annotated to pathways related to the metabolism of carbohydrates, FA, and amino acid in the PP period. The DAP between FA and the interaction effect were annotated to the metabolism of xenobiotics by cytochrome P450, drug metabolism - cytochrome P450, retinol metabolism, and steroid hormone biosynthesis. Collectively, ASCA provided novel information on molecular markers of metabolic adaptations and their interactions with EFA + CLA supplementation. Bioinformatics analysis suggested that supplemental EFA + CLA amplified hepatic FA oxidation; cytochrome P450 was enriched to maintain metabolic homeostasis by oxidation/detoxification of endogenous compounds and xenobiotics. SignificanceThis report is among the first ones applying repeated measurement analysis of variance–simultaneous component analysis (ASCA) to deal with longitudinal proteomics results. ASCA separately identified differentially abundant proteins (DAP) in ‘transition time’, ‘between fatty acid treatments’, and ‘their interaction’. We first identified the molecular signature of hepatic metabolic adaptations during postpartum negative energy balance; the enriched pathways were well-known pathways related to mobilizing fatty acids (FA) and amino acids to support continuous energy production through fatty acid oxidation, TCA cycle, and gluconeogenesis. Some of the DAP were not previously reported in transition dairy cows. Secondly, we provide novel information on the mechanisms by which supplemented essential FA and conjugated linoleic acids interact with hepatic metabolism. In this regard, FA amplified hepatic detoxifying and oxidation capacity through ligand activation of nuclear receptors. Finally, we briefly compared the strengths and weaknesses of the ASCA model with PLS-DA and outlined why these methods are complementary.

Analysis of the transcriptome of bovine endometrial cells isolated by laser micro-dissection (2): impacts of post-partum negative energy balance on stromal, glandular and luminal epithelial cells

BackgroundIn post-partum dairy cows, the energy needs to satisfy high milk production induces a status of more or less pronounced Negative Energy Balance (NEB). NEB associated with fat mobilization impairs reproductive function. In a companion paper, we described constitutive gene expression in the three main endometrial cell types (stromal, glandular and luminal epithelial cells) isolated by laser capture micro-dissection (LCM) showing the specificities of their transcriptomic profiles. This study investigates the specific impact of NEB on gene expression in these cells around 80 days after parturition at day 15 of the oestrus cycle and describes their specific response to NEB.ResultsFollowing the description of their constitutive expression, the transcriptome profiles obtained by RNA sequencing of the three cells types revealed that differences related to the severity of NEB altered mainly specific patterns of expression related to individual cell types. Number of differentially expressed genes between severe NEB (SNEB) and mild NEB (MNEB) cows was higher in ST than in LE and GE, respectively. SNEB was associated with differential expression of genes coding for proteins involved in metabolic processes and embryo-maternal interactions in ST. Under-expression of genes encoding proteins with functions related to cell structure was found in GE whereas genes encoding proteins participating in pro-inflammatory pathways were over-expressed. Genes associated to adaptive immunity were under-expressed in LE.ConclusionThe severity of NEB after calving is associated with changes in gene expression around 80 days after parturition corresponding to the time of breeding. Specific alterations in GEs are associated with activation of pro-inflammatory mechanisms. Concomitantly, changes in the expression of genes encoding proteins involved in cell interactions and maternal recognition of pregnancy takes place in ST. The combination of these effects possibly altering the uterine environment and embryo maternal interactions may negatively influence the establishment of pregnancy.

Association of blood calcium concentration in the first 3 days after parturition and energy balance metabolites at day 3 in milk with disease and production outcomes in multiparous Jersey cows

Research exploring specific associations of markers of negative energy balance and Ca in postpartum Jersey cows with lactation performance is lacking. Our objectives were to evaluate the associations of total Ca concentration (tCa) measured at 1 through 3 d in milk (DIM) and free fatty acids (FFA), β-hydroxybutyrate (BHB), and glucose measured at 3 DIM with (1) the risk of multiparous Jersey cows being diagnosed with early-lactation diseases and culling, (2) milk production in the first 9 wk of lactation, and (3) the risk of pregnancy in the first 150 DIM. A cohort study was performed in 1 dairy herd in Texas. Multivariable Poisson regression models were built to evaluate the association of the analytes of interest with the risks of early-lactation diseases and culling in the first 60 DIM (i.e., binary outcomes). Linear mixed models were used to evaluate the association of the analytes of interest with milk production within the first 9 wk of lactation, and a Cox proportional hazard model was built to assess the risk of pregnancy within 150 DIM. A total of 380 cows were used in the final analyses. Total Ca measured at 1 through 3 DIM was not associated with the risk of metritis. Cows with increased FFA and BHB had an increased risk of being diagnosed with metritis and clinical mastitis, respectively. Increased concentrations of glucose and FFA and decreased tCa at 3 DIM were associated with an increased risk of culling. Reduced tCa concentrations at 1 DIM (≤1.84 mmol/L) and 2 DIM (≤2.04 mmol/L) were associated with increased milk production across the first 9 wk of lactation compared with tCa concentrations above those thresholds. Total Ca was not associated with milk production when assessed at 3 DIM, whereas increased FFA (≥0.37 mmol/L) and decreased glucose (≤2.96 mmol/L) at 3 DIM were associated with increased milk production. None of the metabolites measured were associated with the risk of pregnancy in the first 150 DIM. Our results demonstrate that tCa concentration assessed in the first 3 DIM show temporary associations with milk production and culling in multiparous Jersey cows. Although increased concentration of FFA assessed at 3 DIM was associated with greater milk yield, it was a detrimental factor for the risk of metritis. This study attempted to better elucidate the relationship of tCa, FFA, BHB, and glucose assessed in early postpartum with health and performance of Jersey cows. Based on this study, assessments performed at 3 DIM using tCa concentration ≤1.99 mmol/L for increased risk of early-lactation culling and FFA ≥0.43 mmol/L for increased risk of metritis could be used as starting points. More studies evaluating the dynamics of energy balance markers and tCa in postpartum Jersey cows using a greater number of herds are needed to better inform dairy consultants on critical levels for exacerbated postpartum negative energy balance and subclinical hypocalcemia for the Jersey breed.

Follicular fluid proteomic profiling of dairy cows with anestrus caused by negative energy balance

Negative energy balance (NEB) is an important risk factor for the dairy cow’s anoestrus. The purpose of the experiment is to screen out differentially expressed proteins (DEPs) in the follicular fluid (FF) of cows with anoestrus caused by NEB and oestrus from positive energy balance (PEB). NEB and PEB holstein cows were selected 14–21 days postpartum according to the serum concentrations of β-hydroxybutyric acid (BHB) and glucose (Glu). Based on the follicle diameter at 55–60 days postpartum, the two groups were assigned into the anoestrus group (NEB-A; n = 6; follicular diameter < 8 mm) and oestrus group (PEB-E; n = 6; follicular diameter: 15–20 mm). Their FF was aspirated in vivo to screen out DEPs, and analysed by bioinformatics. Five DEPs were verified by western blotting (WB). The study identified 367 proteins. Compared with PEB-E, NEB-A expressed 135 downregulated DEPs and 37 upregulated DEPs. RBP4 (retinol-binding protein 4), adiponectin B, SOD (superoxide dismutase), IGF2 (insulin-like growth factor 2), and CRP (C-reactive protein) were verified by WB, their may have confirmed that cellular lipid metabolism, oxidative stress, and cellular immune function are related to follicular development and anoestrus. Highlights 367 proteins identified in the Follicle fluid of dairy cows 55–60 days postpartum. In the Negative energy balance-anoestrus group, 172 differentially expressed proteins were identified—135 upregulated DEPs and 37 downregulated DEPs. Confirmed that postpartum Negative energy balance may affect cows’ oestrus behaviour or performance during early lactation by lipid metabolism, the IGF regulation system, and immune and complement systems.

Impacts of feeding a fish-oil based feed supplement through 160 days in milk on reproductive and productive performance, as well as the health, of multiparous early-lactation Holstein cows

A major cause of reproductive failure in contemporary lactating dairy cows is early post-partum negative energy balance (NEB). A strategy to alleviate NEB is to reduce milk energy density by feeding supplements to suppress milk fat synthesis, which could reduce body fat mobilization to favor earlier ovulation and improved pregnancy rates. As fish oils (FO) can cause milk fat depression, and are rich in n-3 PUFA which are beneficial to reproduction, our objective was to evaluate impacts of feeding the FO based feed supplement ‘Optomega Plus’ (OP+) on production, EB, reproduction and health of multiparous Holstein cows through ∼160 days in milk (DIM). Two pens, each of ∼310 multiparous early lactation cows, were fed rations formulated to deliver 0 (C) or 77 g/cow per day of FO (2.53 g/kg DM) as OP+, which contains 3.9 g/cow per day of eicosapentaenoic acid (EPA; C20:5 n-3) and 7.2 g/cow per day of docosahexaenoic acid (DHA; C22:6 n-3). Milk yields and components were recorded monthly for five months after cows were assigned to treatments from a common fresh cow pen at 13.3 ± 0.19 DIM, and body condition (BCS) was scored monthly on a subset of the cows. Dry matter intake (pen basis) was unaffected by treatment, while milk yield, milk components and BCS all had treatment*DIM interactions (P < 0.05), which suggested different responses to the diets before and after ∼130 days post treatment assignment. Feeding OP+ did not impact milk production, but moderately reduced (P < 0.05) milk fat output in the earlier period, which mitigated the NEB as judged by BCS change, but 1st service pregnancy rate was unaffected. There were no treatment impacts on indices of cow health, but feeding OP + improved the healthfulness of the milk FA profile to its human consumers. Results suggest that OP + as a source of FO modestly reduced milk fat output and the extent of early postpartum NEB, without negatively affecting DM intake, health or reproductive performance, while improving the nutritional value of milk fat to its human consumers.

Mid-infrared spectroscopic analysis of raw milk to predict the blood nonesterified fatty acid concentrations in dairy cows

In high-yielding dairy cattle, severe postpartum negative energy balance is often associated with metabolic and infectious disorders that negatively affect production, fertility, and welfare. Mobilization of adipose tissue associated with negative energy balance is reflected through an increased level of nonesterified fatty acids (NEFA) in the blood plasma. Earlier, identification of negative energy balance through detection of increased blood plasma NEFA concentration required laborious and stressful blood sampling. More recently, attempts have been made to predict blood NEFA concentration from milk samples. In this study, we aimed to develop and validate a model to predict blood plasma NEFA concentration using the milk mid-infrared (MIR) spectra that are routinely measured in the context of milk recording. To this end, blood plasma and milk samples were collected in wk 2, 3, and 20 postpartum for 192 lactations in 3 herds. The blood plasma samples were taken in the morning, and representative milk samples were collected during the morning and evening milk sessions on the same day. To predict plasma NEFA concentration from the milk MIR spectra, partial least squares regression models were trained on part of the observations from the first herd. The models were then thoroughly validated on all other observations of the first herd and on the observations of the 2 independent herds to explore their robustness and wide applicability. The final model could accurately predict blood plasma NEFA concentrations <0.6 mmol/L with a root mean square error of prediction of <0.143 mmol/L. However, for blood plasma with >1.2 mmol/L NEFA, the model clearly underestimated the true level. Additionally, we found that morning blood plasma NEFA levels were predicted with significantly higher accuracy using MIR spectra of evening milk samples compared with MIR spectra of morning samples, with root mean square error of prediction values of, respectively, 0.182 and 0.197 mmol/L, and R2 values of 0.613 and 0.502. These results suggest a time delay between variations in blood plasma NEFA and related milk biomarkers. Based on the MIR spectra of evening milk samples, cows at risk for negative energy status, indicated by detrimental morning blood plasma NEFA levels (>0.6 mmol/L), could be identified with a sensitivity and specificity of, respectively, 0.831 and 0.800. As this model can be applied to millions of historical and future milk MIR spectra, it opens an opportunity for regular metabolic screening and improved resilience phenotyping.

Milk yield at first lactation, parity, and season of calving affect the reproductive performance of water buffalo cows

Context Profitability of water buffalo systems depends on a calving interval (CI) &amp;lt;400 days. Several factors affect the achievement of this target. However, milk yield at first lactation has received little attention. Aims Determine the effect of milk yield at first lactation, parity, season of calving and farm, on the length of CI and the probability of a CI ≤ 400 days in water buffaloes. Methods A retrospective analysis of milk yield at first lactation and reproductive records of 1459 water buffaloes was carried out. Milk yield was categorised as Group 1 (≤1090 kg), Group 2 (1090–1377 kg), Group 3 (1377–1684 kg) and Group 4 (&amp;gt;1684 kg); parity was categorised as parity 1, 2 and ≥ 3; and month of calving was grouped into three seasons: December–March, April–July, and August–November. Data were analysed using linear and logistic mixed models. Key results CI increased from 425.3 days (95% CI: 418.8–431.8 days) in group 1 to 463.3 days (95% CI: 456–470.6 days) in group 4 (P &amp;lt; 0.05), while the probability of having a CI ≤ 400 days decreased from 0.5 (95% CI: 0.46–0.54) to 0.26 (95% CI: 0.22–0.29), respectively (P &amp;lt; 0.05). CI decreased from 466 days (95% CI: 460.8–471.3 days) in parity 1 to 410.5 days (95% CI: 405.2–415.8 days) in parity ≥3, whereas the probability of a CI ≤ 400 days increased from 0.26 (95% CI: 0.24–0.29) to 0.51 (95% CI: 0.47–0.54) respectively (P &amp;lt; 0.05). Water buffaloes calving in August–November showed significantly shorter CI and, along with those calving between December–March, showed the highest probability of a CI ≤ 400 days. An interaction between milk yield at first lactation and parity on both outcomes was observed. Conclusions Shorter CI and higher probability of a CI &amp;lt;400 days were associated with lower milk yields at first lactation, higher parity and calving between August–November. Higher milk yield at first lactation affected negatively the reproductive performance of water buffaloes, especially at parity 1 and 2. Implications These results highlight the importance of adequate nutritional management to allow water buffaloes to cope with the challenge of the postpartum negative energy balance and have a calving interval less than 400 days.

Extracellular vesicle-coupled miRNA profiles in follicular fluid of cows with divergent post-calving metabolic status

Most high-yielding dairy cows enter a state of negative energy balance (NEB) during early lactation. This, in turn, results in changes in the level of various metabolites in the blood and follicular fluid microenvironment which contributes to disturbed fertility. Extracellular vesicles (EVs) are evolutionarily conserved communicasomes that transport cargo of miRNA, proteins and lipids. EV-coupled miRNAs have been reported in follicular fluid. However, the association between postpartum NEB and EV-coupled miRNA signatures in follicular fluid is not yet known. Energy balance analysis in lactating cows shortly after post-calving revealed that the majority of the cows exhibited transiently negative energy balance levels, whereas the remaining cows exhibited either consistently negative or consistently positive energy levels. Metabolic status was associated with EV-coupled miRNA composition in the follicular fluid. Cows experiencing NEB showed reduced expression of a large number of miRNAs while cows with positive energy balances primarily exhibited elevated expression of EV-coupled miRNAs. The miRNAs that were suppressed under NEB were found to be involved in various metabolic pathways. This is the first study to reveal the presence of an association between EV-coupled miRNA in follicular fluid and metabolic stress in dairy cows. The involvement of differentially expressed miRNAs in various pathways associated with follicular growth and oocyte maturation suggest the potential involvement of specific follicular miRNAs in oocyte developmental competence, which may partially explain reduced fertility in cows due to post-calving metabolic stress.

Sensor based eating time variables of dairy cows in the transition period related to the time to first service

In dairy cattle, reproductive diseases and infertility are some of the most important reasons for culling, where postpartum negative energy balance (NEB) reduces reproductive performance. This single cohort observational study reports the association between eating time and the interval between calving and first service in 2036 dairy cows on 17 commercial farms in The Netherlands. Cows were equipped with a commercially available neck sensor (Nedap, Groenlo, The Netherlands), that measured the time cows spent eating, from 28 days (d) before until 28 d after parturition. Primiparous cows spent a mean of +45 minutes (min) eating time per day ante partum and +15 min eating time post partum more than multiparous cows. A Cox proportional hazard model was used to analyze eating time variables in relation to the interval between calving and first service. From 4 weeks before until 4 weeks after calving eating time variables per week were used. Weeks -4, -3 + 3 and +4 were used as weeks with stable eating time patterns and therefore the mean eating time per week and the standard deviation of the mean eating time per week were used. Weeks -2, -1, +1 and +2 were addressed as periods with unstable eating patterns and therefore the slope in eating time per week and the residual variance of the slope per week were modeled. Significant results were the mean eating time in week -4 and +3 where in both weeks higher eating time lead to a higher hazard for first service. Difference between primiparous and multiparous cows were also significant with a higher hazard for first service for primiparous cows. Week 4 post partum presented a significant difference between eating time of primiparous cows and multiparous cows. These results display how eating time variables in the transition period could be related to the interval between calving and first service, and that there is a relation between mean eating time in week -4, +3, +4 and the interval between calving and first insemination.

Buffalo liver transcriptome analysis suggests immune tolerance as its key adaptive mechanism during early postpartum negative energy balance.

Females undergo negative energy balance (NEB) during the early postpartum period to meet the lactation demands. The liver, being the key metabolic organ, plays a major role in handling NEB. Dairy animals handling high lactation demands are better models to understand the liver adaptive mechanisms during this phase. Therefore, we analyzed the liver transcriptome of dairy buffaloes during early postpartum. Liver biopsies were performed on three lactating buffaloes on the 15th and the 30th days of early postpartum and three heifers (controls) at the diestrous stage. Paired-end Next Generation Sequencing (NGS) identified 509 significantly differentially expressed genes (SDE) in the liver among the three groups. The SDE with log2 fold change > 3 and the unique SDE revealed the promotion of immune suppression (e.g., TCR), apoptosis (e.g., CCDC103), PGF2α synthesis, fat accumulation (e.g., BGLAP) and liver regeneration (e.g., FGF10) pathways, and the downregulation of antigen presentation (e.g., BOLA-DQA) on the 15th day of lactation. Consistently upregulated genes on the 15th and 30th days of early postpartum indicated the promotion of immune tolerance (e.g., IFITM3), medium and long-chain fatty acids' oxidation (e.g., ACSM3), and lipid accumulation (e.g., INSIG1). However, consistently downregulated genes during early postpartum showed immunosuppression, the downregulation of gluconeogenesis from amino acids (e.g., DDO), and the biosynthesis of taurine (e.g., CSAD) and unsaturated fatty acids (e.g., SCD). Functional annotation and network analyses also indicated the promotion of immune tolerance, fat accumulation and decreased gluconeogenesis from amino acids, and estrogen metabolism on the 15th day of lactation. Overall, the liver showed immune tolerance as an adaptive mechanism during early postpartum of buffaloes.

Comprehensive analysis of blood cells and plasma identifies tissue-specific miRNAs as potential novel circulating biomarkers in cattle

BackgroundThe potential of circulating miRNAs as biomarkers of tissue function, both in health and disease, has been extensively demonstrated in humans. In addition, circulating miRNA biomarkers offer significant potential towards improving the productivity of livestock species, however, such potential has been hampered by the absence of information on the nature and source of circulating miRNA populations in these species. In addition, many miRNAs originally proposed as robust biomarkers of a particular tissue or disease in humans have been later shown not to be tissue specific and thus to actually have limited biomarker utility. In this study, we comprehensively analysed miRNA profiles in plasma and cell fractions of blood from cattle with the aim to identify tissue-derived miRNAs which may be useful as biomarkers of tissue function in this important food animal species.ResultsUsing small RNA sequencing, we identified 92 miRNAs with significantly higher expression in plasma compared to paired blood cell samples (n = 4 cows). Differences in miRNA levels between plasma and cell fractions were validated for eight out of 10 miRNAs using RT-qPCR (n = 10 cows). Among miRNAs found to be enriched in plasma, we confirmed miR-122 (liver), miR-133a (muscle) and miR-215 (intestine) to be tissue-enriched, as reported for other species. Profiling of additional miRNAs across different tissues identified the human homologue, miR-802, as highly enriched specifically in liver.ConclusionsThese results provide novel information on the source of bovine circulating miRNAs and could significantly facilitate the identification of production-relevant tissue biomarkers in livestock. In particular, miR-802, a circulating miRNA not previously identified in cattle, can reportedly regulate insulin sensitivity and lipid metabolism, and thus could potentially provide a specific biomarker of liver function, a key parameter in the context of post-partum negative energy balance in dairy cows.

Thyroid Hormones, Insulin, Body Fat, and Blood Biochemistry Indices in Dairy Cows During the Reproduction/Production Cycle

AbstractThis study investigated the changes in: thyroid hormones, amount of subcutaneous fat, and selected indices of blood biochemistry in dairy cows in relation to the reproduction/production cycle. The blood samples were collected bothante- andpost-partumevery two weeks. When evaluating the mean values of the investigated indices, the major changes were recorded in dairy cows 3 to 14 days after calving. During this period, we observed a significant decrease in the mean serum levels of T3(P &lt; 0.05), T4(P &lt; 0.01), and triglycerides (P &lt; 0.01). An opposite trend was observed with a significant increase after calving in the: mean serum levels of β-hydroxybutyrate (P &lt; 0.05), urea (P &lt; 0.01), and mean AST activities (P &lt; 0.05). A significant increase over the normal range was recorded in the average levels of non-esterified fatty acids (P &lt; 0.01) and total bilirubin (P &lt; 0.01). From the next sampling (28 days after calving) onwards we recorded a significant increase in the blood serum levels of cholesterol (P &lt; 0.01), total lipids (P &lt; 0.01), total protein (P &lt; 0.01), as well as a significant decrease in the insulin levels (P &lt; 0.05) and a reduced layer of subcutaneous fat (P &lt; 0.01). The blood serum iodine concentration showed only slight significant changes (P &lt; 0.05) during the observation. Blood serum levels of glucose did not show any significant changes during the whole observation period. Within the whole observation period we found a negative correlation between T3levels and the layer of subcutaneous fat (r = −0.2606; P &lt; 0.05). This correlation was much more marked in cows 3 to 14 days after calving (r = −0.5077; P &lt; 0.05), which may indicate a possible relationships between the thyroid status, body condition, andpost partumnegative energy balance.

Investigation of negative energy balance and postpartum anoestrus in an intensive dairy farm from the Chinese province of Heilongjiang

The aim of this investigation was to determine the relationship between postpartum anoestrus and negative energy balance in an intensive dairy farm from the Heilongjiang Province, China. At 14 to 21 d after parturition, 100 cows were randomly selected and their plasma indices, including β-hydroxybutyric acid, non-esterified fatty acid, and glucose were measured. Cows were assigned to a positive energy balance group (n = 37) and a negative energy balance group (n = 36) based on their β-hydroxybutyric acid concentrations (&gt; 1.20 mmol/l). The two groups of cows were examined by B-mode ultrasonography and rectal examination from 60 to 90 d after parturition to identify the ovarian status of oestrous and anoestrous animals. The incidences of negative energy balance and positive energy balance were 49 and 57%, respectively, from14 to 21 d after parturition. From 60 to 90 d after parturition, 94.4% of the negative energy balance group were in anoestrus and 5.6% were in oestrus, while 62.2% of the positive energy balance group were in anoestrus and 37.8% were in oestrus. Furthermore, the proportion of inactive ovaries in the negative energy balance group was 61.8%. In conclusion, the negative energy balance is an important factor causing inactive ovaries in high-yielding dairy cows.

Effects of Dietary Energy Density in the Dry Period on the Production Performance and Metabolism of Dairy Cows

Thirty healthy Holstein dairy cows in the dry period were randomly divided into three groups and fed diets with different net energy for lactation (group A: 1.2 Mcal/kg DM, group B: 1.3 Mcal/kg DM, and group C: 1.4 Mcal/kg DM) for 8 weeks prepartum. Thereafter, dairy cows were fed a diet of the same formulation (1.66 Mcal/kg DM) for 12 weeks postpartum. The effects of different dietary energy densities in the dry period on postpartum performance and metabolic parameters of dairy cows were observed. Milk yield was reduced by 14.5% in the low-energy diet group; however, there were no differences in milk composition between the three groups. Postpartum plasma β-hydroxybutyrate, non-esterified fatty acid, growth hormone, and glucagon levels were significantly decreased whereas leptin and neuropeptide levels were elevated in the low-energy diet group. Moreover, body fat mobilization was attenuated, and the decline in postpartum body condition was reduced in the low-energy diet group, thus effectively reducing the postpartum negative energy balance.

Prepartum body condition score and plane of nutrition affect the hepatic transcriptome during the transition period in grazing dairy cows.

BackgroundA transcriptomic approach was used to evaluate potential interactions between prepartum body condition score (BCS) and feeding management in the weeks before calving on hepatic metabolism during the periparturient period.MethodsThirty-two mid-lactation grazing dairy cows of mixed age and breed were randomly allocated to one of four treatment groups in a 2 × 2 factorial arrangement: two prepartum BCS categories [4.0 (thin, BCS4) and 5.0 (optimal, BCS5); based on a 10-point scale], and two levels of energy intake during the 3 weeks preceding calving (75 and 125 % of estimated requirements). Liver samples were obtained at −7, 7, and 28 d relative to parturition and subsequent RNA was hybridized to the Agilent 44 K Bovine (V2) Microarray chip. The Dynamic Impact Approach was used for pathway analysis, and Ingenuity Pathway Analysis was used for gene network analysis.ResultsThe greater number of differentially expressed genes in BCS4 cows in response to prepartum feed allowance (1071 vs 310, over the entire transition period) indicates that these animals were more responsive to prepartum nutrition management than optimally-conditioned cows. However, independent of prepartum BCS, pathway analysis revealed that prepartal feeding level had a marked effect on carbohydrate, amino acid, lipid, and glycan metabolism. Altered carbohydrate and amino acid metabolism suggest a greater and more prolonged negative energy balance postpartum in BCS5 cows overfed prepartum. This is supported by opposite effects of prepartum feeding in BCS4 compared with BCS5 cows in pathways encompassing amino acid, vitamin, and co-factor metabolism. The prepartum feed restriction ameliorates the metabolic adaptation to the onset of lactation in BCS5 cows, while detrimentally affecting BCS4 cows, which seem to better adapt when overfed. Alterations in the glycosaminoglycans synthesis pathway support this idea, indicating better hepatic health status in feed-restricted BCS5 and overfed BCS4 cows. Furthermore, IPA network analysis suggests liver damage in feed-restricted thin cows, likely due to metabolic overload.ConclusionOverall, the data support the hypothesis that overfeeding in late-pregnancy should be limited to underconditioned cows, while cows with optimal degree of body condition should be maintained on an energy-restricted diet.Electronic supplementary materialThe online version of this article (doi:10.1186/s12864-016-3191-3) contains supplementary material, which is available to authorized users.

Insulin signaling, inflammation, and lipolysis in subcutaneous adipose tissue of transition dairy cows either overfed energy during the prepartum period or fed a controlled-energy diet

Adipose tissue mobilization is a hallmark of the transition period in dairy cows. Cows overfed energy during the dry period have higher concentrations of nonesterified fatty acids (NEFA) and β-hydroxybutyrate (BHB) compared with cows fed a controlled-energy diet prepartum. The reason for an increase in blood NEFA concentrations at the level of adipose tissue in cows overfed energy has not been fully elucidated. One hypothesis is that cows with high BHB concentrations suffer from adipose tissue-specific insulin resistance, leading to higher rates of adipose tissue mobilization in the postpartum period. To test this hypothesis, subcutaneous adipose tissue biopsies of cows overfed energy in excess of predicted requirements by 50% in the dry period, and that had high concentrations of blood BHB postpartum (group H; n=12), were used. Findings were compared with results of biopsies from cows fed a controlled-energy diet and with low BHB concentrations postpartum (group C; n=12) to create the biggest contrast in BHB concentrations. Subcutaneous adipose tissue biopsies were obtained before and 60 min after an intravenous glucose challenge (0.25 g/kg of glucose) at 28 and 10 d before expected calving as well as on d 4 and 21 postpartum. Phosphorylation of protein kinase B, extracellular signal-regulated kinase, and hormone-sensitive lipase was determined before and after glucose infusion by Western blot. Western blot was also used to assess the baseline protein abundance of peroxisome proliferator-activated receptor gamma and insulin receptor β-subunit. In addition, gene expression of fatty acid synthase, adiponectin, monocyte chemoattractant protein 1, and tumor necrosis factor α was determined by real-time quantitative reverse-transcription PCR. Backfat thickness was determined in the thurl area by ultrasonography. Cows in group H showed a greater degree of lipogenesis prepartum, but no differences were found in lipolytic enzyme activity postpartum compared with cows in group C. Baseline plasma insulin concentrations were decreased and serum NEFA concentrations increased postpartum in group H. Insulin signaling through protein kinase B, quantity of insulin receptor, markers of inflammation, and peroxisome proliferator-activated receptor gamma in adipose tissue were not different between the groups, but expression of adiponectin was increased in adipose tissue of cows in group H during the immediate peripartum period. In conclusion, differences in serum concentrations of NEFA between cows overfed energy prepartum and high blood concentrations of BHB are likely due to greater negative energy balance postpartum reflected in lower circulating concentrations of glucose and insulin and an increase in the total amount of mobilized adipose tissue mass rather than due to changes in adipose tissue insulin signaling.

Insulin signaling and skeletal muscle atrophy and autophagy in transition dairy cows either overfed energy or fed a controlled energy diet prepartum.

During periods of negative energy balance, mobilization of muscle is a physiologic process providing energy and amino acids. This is important in transition dairy cows experiencing negative energy and protein balance postpartum. Overconsumption of energy during late pregnancy affects resting glucose and insulin concentrations peripartum and increases the risk for hyperketonemia postpartum, but the effects on muscle tissue are not fully understood. Skeletal muscle accounts for the majority of insulin-dependent glucose utilization in ruminants. Our objective was to study peripartal skeletal muscle insulin signaling as well as muscle accretion and atrophy in cows with excess energy consumption prepartum. Skeletal muscle biopsies were obtained 28 and 10 days prepartum, as well as 4 and 21 days postpartum from 24 Holstein cows. Biopsies were taken immediately before and 60 min after intravenous glucose challenge causing endogenous release of insulin. Gene expression of IGF-1, myostatin, and atrogin-1, as well as immunoblot analysis of atrogin-1, muRF1, ubiquitinated proteins, LC3, and phosphorylation of AKT, ERK and mTORC1 substrate 4EBP1 was performed. Excess energy consumption in late pregnancy did not lead to changes in insulin-dependent molecular regulation of muscle accretion or atrophy compared with the controlled energy group. In both groups, phosphorylation of AKT and mTORC1 substrate was significantly decreased postpartum whereas proteasome activity and macroautopagy were upregulated. This study showed that in addition to the proteasome pathway of muscle atrophy, macroautophagy is upregulated in postpartum negative energy and protein balance regardless of dietary energy strategy prepartum and was higher in cows overfed energy throughout the study period.

Nutrition, Metabolic Status and Reproductive Efficiency in Dairy Herds

Nutrition of transition period in dairy cows dramatically increases requirements for energy, glucose, amino acids and other nutrients in dairy cows and, parallel to that, feed intake is depressed. An insufficient or imbalanced nutrient intake might lead to the malfunction of organs, causing reproductive failure. To minimize the metabolic/reproductive damage caused by negative energy balance (NEBAL), feeding of a glucogenic and lipogenic diet containing fat-enriched supplements is recommended. In heat-stressed dairy cows, the reduction of dry matter intake causes an energy deficit, thereby accentuating NEBAL. Heat stress may also have a direct impact on oocyte quality due to the high ovarian temperatures. The intrauterine environment is also compromised in heat-stressed cows, with reduced blood flow to the uterus and increased uterine temperature; this may impair embryonic development, increase early embryonic loss and reduce the proportion of successful inseminations. Postpartum NEBAL suppresses immune function and promotes metabolic disorders, potentially explaining relationships between infectious and noninfectious transition disorders. In postpartum dairy cows, pathologic ovarian phenomena including ovulatory (cystic corpora lutea) and non-ovulatory (follicular and luteal cysts) forms frequently occur causing decreased fertility. The main objective of this review was summarizing the most relevant information regarding nutrition and reproduction in dairy cows.