Over-conditioned Cows Research Articles

Prepartum anti-inflammatory therapies in Holstein dairy cows blocked by parity and body condition score group: Effects on metabolic stress, systemic inflammation, performance, and health

The objective of this study was to assess the effects of prepartum administration of anti-inflammatory therapies on body condition score (BCS), β-hydroxybutyrate (BHB) concentration, haptoglobin (HP) concentration, milk yield, milk components, rumination time, clinical health events and reproductive performance in Holstein dairy cows. At 14 d before the expected calving date, cows (PAR; n = 170) and heifers (nulliparous [NUL]; n = 63) were blocked by BCS group (optimal = 3-3.5 [OPT]; over-conditioned cows [OVERC; BCS ≥ 3.75 pts.]) and parity (NUL; PAR) and randomly allocated to one of 3 treatment groups: 1) ASA (n = 78): receive one oral administration of acetylsalicylic acid (4 boluses; 480 grain/bolus); 2) MEL (n = 76): receive one oral administration with meloxicam (1mg/kg of BW), or 3) PLC (n = 77): receive one oral treatment with gelatin capsules filled with water. Body condition score was assessed, and blood samples were collected, weekly starting one week before treatment until 3 weeks after calving. Daily milk yields and daily rumination times were collected from on-farm computer records. Dairy Herd Improvement Association (DHIA) monthly test data were collected to assess milk yield, somatic cell counts, and milk components. Furthermore, health events, culling rate, and reproductive performance data were collected from on-farm computer records. The data were analyzed using MIXED, GLIMMIX, and LIFETEST procedures of SAS as a randomized complete block design. On average, MEL-NUL cows produced 4.77 ± 0.93 kg/d and 4.81 ± 0.92 kg/d more milk from wk 6 to wk 21 of lactation compared with ASA-NUL and PLC-NUL cows, respectively. Similarly, there was a week by treatment by body condition group interaction (P = 0.01), where OVERC cows treated with MEL produced more milk from wk 10 to wk 15 of lactation compared with ASA- OVERC and PLC-OVERC cows. Parous cows treated with ASA had lower BCS compared with PAR cows treated with MEL or PLC. A lower percentage of OVERC cows treated with ASA became sick in the first 60 DIM compared with MEL- OVERC and PLC- OVERC cows (ASA = 23.88 ± 7.26%, MEL = 46.36 ± 8.57%; PLC = 46.74 ± 8.53%; P = 0.04). Parous cows treated with ASA had (P = 0.03) a higher hazard ratio to become pregnant by 300 DIM compared with PAR MEL cows. Although the study was not sized for finding treatment differences in blocking criteria groups, these results suggest that treatment with prepartum anti-inflammatory therapies may have positive effects on milk yield and postpartum health in specific groups of cows, such as NUL and OVERC cows, while it may not be recommended for other animal categories, such as parous cows and cows with optimal BCS. Larger studies are needed to strengthen the associations observed in this study.

Liver transcriptome profiles of dairy cows with different serum metabotypes

In a previously established animal model, 38 multiparous Holstein cows were assigned to 2 groups fed different diets to achieve either a normal (NBCS) or high (HBCS) body condition score (BCS) and backfat thickness (BFT) until dry-off at -49 d before calving (NBCS: BCS <3.5 [3.02 ± 0.24) and BFT <1.2 cm [0.92 ± 0.21]; HBCS: BCS >3.75 [3.82 ± 0.33] and BFT >1.4 cm [2.36 ± 0.35], mean ± SD). The groups were also stratified for comparable milk yields (NBCS: 10,361 ± 302 kg; HBCS: 10,315 ± 437 kg; mean ± SD). The cows were then fed the same diet during the dry period and subsequent lactation, maintaining the differences in BFT and BCS throughout the study. Using the serum metabolomics data, we created a classification model that identified different metabotypes. Machine learning classifiers revealed a distinct cluster labeled HBCS-PN (HBCS predicted normal BCS) among over-conditioned cows. These cows showed higher feed intake and better energy balance than the HBCS-PH (high BCS predicted high BCS) group, while milk yield was similar. The aim of this study was to investigate the changes in the hepatic transcriptome of cows differing in serum-metabotype postpartum. We performed hepatic transcriptome analysis in cows from 3 metabolic clusters: HBCS-PH (n = 8), HBCS-PN (n = 6), and normal BCS predicted normal BCS (NBCS-PN, n = 8) on d 21 (±2) postpartum. Liver tissue from cows expressed a total of 13,118 genes aligned with the bovine genome. A total of 48 differentially expressed genes (DEG; false discovery rate ≤0.1 and fold-change >1.5) were found between NBCS-PN and HBCS-PH cows, whereas 24 DEG (14 downregulated and 10 upregulated) were found between HBCS-PN and HBCS-PH cows. The downregulated DEG (n = 31) in NBCS-PN cows compared with HBCS-PH cows are involved in biosynthetic processes such as lipid, lipoprotein, and cholesterol synthesis (e.g., APOA1, MKX, RPL3L, CANT1, CHPF, FUT1, ZNF696), cell organization, biogenesis, and localization (e.g., SLC12A8, APOA1, BRME1, RPL3L, STAG3, FBXW5, TMEM120A, SLC16A5, FGF21), catabolic processes (e.g., BREH1, MIOX, APOBEC2, FBXW5, NUDT16), and response to external stimuli (e.g., APOA1, FGF21, TMEM120A, FNDC4), whereas upregulated DEG (n = 17) are related to signal transduction and cell motility (e.g., RASSF2, ASPN, SGK1, KIF7, ZEB2, MAOA, ACKR4, TCAF1), suggesting altered metabolic adaptations during lactation. Our results showed 24 DEG between HBCS-PN and HBCS-PH in the liver. The expression of SLC12A8, SLC16A5, FBXW5, OSGIN1, LAMA3, KDELR3, OR4X17, and INHBE, which are responsible for regulating cellular processes was downregulated in HBCS-PN cows compared with HBCS-PH cows. In particular, the downregulation of SLC12A8 and SLC16A5 expression in HBCS-PN cows indicates lower metabolic load and reduced need for NAD+ biosynthesis to support mitochondrial respiratory processes. The upregulation of MAOA, ACKR4, KIF27, SFRP1, and CAV2 in the liver of HBCS-PN cows may indicate adaptive mechanisms to maintain normal liver function in response to increased metabolic demands from over-conditioning. These molecular differences underscore the existence of distinct metabolic types in cows and provide evidence for the role of the liver in shaping different metabolic patterns.

The morphological and functional ultrastructure of cells in pre-implantation embryos.

Cells of pre-implantation embryos are equipped with many morphological and functional systems through which they can synthesize specific proteins and effectively ensure the protection of early embryonic development. Here we present evidence for the existence of these systems in morphologically normal and abnormal bovine blastocyst stage embryos in vivo at the ultrastructural and actin cytoskeleton levels. The appearance of organelles in the trophectoderm (TE) and inner cell mass (ICM) cells, responsible for their synthetic activities and their role in the development of early bovine embryos are described. We point out the importance of endocytic processes and the participation of extracellular vesicles in the formation of intercellular contacts and homeostasis of the embryo microenvironment. Several changes in the ultrastructural morphology of embryos produced by different methods (ICSI, parthenogenetic AC/DC electrical activation, IVF with separated sperm) and freezing/thawed embryos are described. We also show alterations occurred in the organelles after viral contamination of embryos with BHV-1 and BVDV viruses, and in embryos from over-conditioned cows. Recorded changes in organelles and appearance of cellular autophagic structures (vesicles, multivesicular bodies and autophagolysosomes) may negatively affect embryo metabolism and lead to the emergence of pathological processes in TE and ICM cells of preimplantation embryos.

Identification and characterization of dairy cows with different backfat thickness antepartum in relation to postpartum loss of backfat thickness: A cluster analytic approach

The objectives of this study were (1) to characterize the interindividual variation in the relationship between antepartum (ap) backfat thickness (BFT) and subsequent BFT loss during early lactation in a large dairy herd using cluster analysis; (2) to compare the serum concentrations of metabolites (nonesterified fatty acids, β-hydroxybutyrate), metabolic hormones (leptin and adiponectin), and an inflammatory marker (haptoglobin) among the respective clusters; and (3) to compare lactation performance and uterine health status in the different clusters. An additional objective was (4) to investigate differences in these serum variables and in milk yield of overconditioned (OC) cows that differed in the extent of BFT loss. Using data from a large study of 1,709 multiparous Holstein cows, we first selected those animals from which serum samples and BFT results (mm) were available at d 25 (±10) ap and d 31 (±3 d) postpartum (pp). The remaining 713 cows (parity of 2 to 7) were then subjected to cluster analysis: different approaches based on the BFT of the cows were performed. K-means (unsupervised machine learning algorithm) clustering based on BFT-ap alone identified 5 clusters: lean (5-8 mm BFT, n = 50), normal (9-12 mm, n = 206), slightly fat (SF; 13-16 mm, n = 203), just fat (JF; 16-22 mm, n = 193), and very fat (VF; 23-43 mm, n = 61). Clustering by difference between BFT-ap and BFT-pp (ΔBFT) also revealed 5 clusters: extreme loss (17-23 mm ΔBFT, n = 16), moderate loss (9-15 mm, n = 119), little loss (4-8 mm, n = 326), no loss (0-3 mm, n = 203), and gain (-8 to -1 mm, n = 51). Based on the blood variables measured, our results confirm that cows with greater BFT losses had higher lipid mobilization and ketogenesis than cows with less BFT loss. The serum variables of cows that gained BFT did not differ from normal cows. Milk yield was affected by the BFT-ap cluster, but not by the ΔBFT cluster. Cows categorized as VF had lesser milk yield than other clusters. We further compared the OC cows that had little or no BFT loss (i.e., 2% of VF, 12% of JF, and 31% of SF, OC-no loss, n = 85) with the OC cows that lost BFT (OC-loss, n = 135). Both NEFA and BHB pp concentrations and milk yield were greater in OC-loss cows compared with the OC-no loss cows. The serum concentration of leptin ap was greater in OC-loss than in the OC-no loss cows. Overall, OC cows lost more BFT than normal or lean cows. However, those OC cows with a smaller loss of BFT produced less milk than OC cows with greater losses.

Farm-level factors associated with lameness prevalence, productivity, and milk quality in farms with automated milking systems

Impaired locomotion (lameness) may negatively affect the ability and desire of cows to milk voluntarily, which is a key factor in success of automated milking systems (AMS). The objective of this study was to identify factors associated with herd-level lameness prevalence and associations of lameness and other farm-level factors with milking activity, milk yield, and milk quality in herds with AMS. From April to September 2019, 75 herds with AMS in Ontario, Canada, were visited, and data on barn design and farm management practices were collected. Data from AMS were collected, along with milk recording data, for the 6-mo period before farm visits. Farms averaged 98 ± 71 lactating cows, 2.3 ± 1.5 robot units/farm, 43.6 ± 9.4 cows/robot, 36.4 ± 4.9 kg/d of milk, a milking frequency of 3.01 ± 0.33 milkings/d, and a herd average geometric mean SCC of 179.3 ± 74.6 (× 1,000) cells/mL. Thirty percent of cows/farm (minimum of 30 cows/farm) were scored for body condition (1 = underconditioned to 5 = over conditioned) and locomotion (1 = sound to 5 = lame; clinically lame ≥3 out of 5 = 28.3 ± 11.7%, and severely lame ≥4 out of 5 = 3.0 ± 3.2%). Clinical lameness (locomotion score ≥3) was less prevalent on farms with sand bedding, with increased feed bunk space per cow, and on farms with non-Holstein breeds versus Holsteins, and tended to be less prevalent with lesser proportion of underconditioned cows (with body condition score ≤2.5). Severe lameness occurrence (farms with any cows with locomotion score ≥4) was associated with a greater proportion of underconditioned cows and in farms with stalls with greater curb heights. Herd average milk yield/cow per day increased with lesser prevalence of clinical lameness (each 10-percentage-point decrease in clinical lameness prevalence was associated with 2.0 kg/cow per day greater milk yield) and greater milking visit frequency per day, and tended to be greater with increased feed push-up frequency. Lesser herd average somatic cell count was associated with lesser clinical lameness prevalence, herd average days in milk, and proportion of overconditioned cows, and somatic cell count tended to be lesser for farms with sand bedding versus those with organic bedding substrates. The results highlight the importance of minimizing lameness prevalence, using of sand bedding, ensuring adequate feed access and feed bunk space, and maintaining proper cow body condition to optimize herd-level productivity and milk quality in AMS herds.

Effect of pegbovigrastimon clinical mastitis and uterine disease during a full lactation in grazing dairy cows.

In this randomized controlled trial on four commercial grazing dairy farms, we investigated whether pegbovigrastim (PEG) treatment affects clinical mastitis (CM) and uterine disease (i.e. retained placenta (RP), metritis and endometritis) occurrence during a full lactation. The association of prepartum body condition score and prepartum non-esterified fatty acid (NEFA) concentration with disease occurrence was also evaluated. Holstein cows were randomly assigned to one of two treatments: first PEG dose approximately 7 d before the expected calving date and a second dose within 24 h after calving (PEG) compared to untreated controls (Control). In total, 2,153 animals were included in the study: 733 primiparous cows (Control = 391, PEG = 342) and 1420 multiparous cows (Control = 723, PEG = 697). Treatment effects were evaluated with generalized linear mixed models and Cox’s proportional hazard models. Treatment with PEG reduced the occurrence of a first case of CM during the first 30 days in milk (DIM) by 24.6% and reduced the hazard of a first case and the rate of total cases of CM during the full lactation. All PEG treatment effects were independent of parity. Prepartum body condition score interacted with PEG treatment: in over-conditioned cows, PEG reduced the occurrence of a first case of CM during the first 30 DIM by 49.5%. The hazard analysis of a first case of CM during the full lactation suggested that the preventive effect of PEG disappeared with increasing DIM. Treatment with PEG did not affect the occurrence of RP or metritis. Pegbovigrastim treated cows with metritis subsequently showed a reduced occurrence of endometritis compared to control cows with metritis. Pegbovigrastim reduces the occurrence of CM particularly in cows at risk of elevated lipid mobilization, and PEG ameliorates the uterine healing process in cows that experienced metritis.

Assessing dairy cow welfare during the grazing and housing periods on spring-calving, pasture-based dairy farms.

The different periods characterizing spring-calving, pasture-based dairy systems common in Ireland have seldom been the focus of large-scale dairy cow welfare research. Thus, the aim of this study was to devise and conduct an animal-based welfare assessment during both the grazing and housing periods on spring-calving, pasture-based dairy farms, to identify areas for improvement and establish benchmarks for indicators of good welfare. Assessment of seven animal-based welfare indicators was conducted during two visits (one each at grazing and housing) to 82 commercial dairy farms in southern Ireland. Herd-level descriptive statistics were performed for all welfare indicators at each visit, and differences between visits were analyzed using paired t-tests and Wilcoxon signed-rank tests. A mean of 9% and 10% clinically lame cows (mobility scores 2 and 3) were observed at housing and grazing, respectively. Recommended body condition scores (BCS) were not met for a mean of 13% of cows at grazing and 23% at housing, with more over-conditioned cows present at housing than grazing (P < 0.001). Ocular discharge was uncommon in both periods. Prevalence of moderate and severe nasal discharge combined was lower during housing (5%) than grazing (7%). In both periods, similar mean levels of tail injury were observed: 2% to 3% of cows with tail lacerations, 9% with broken tails, and 8% (measured at housing only) with docked tails. Integument alterations involved primarily hair loss and were most prevalent on the hindquarters (26%) during grazing and on the head–neck–back (66%) and the hindquarter (32%) regions during housing. Cows displayed an avoidance distance of >1 m (indicative of a fearful response) from an approaching human in an average of 82% of grazing cows and 42% to 75% of housed cows, dependent on test location. Opportunities to improve welfare in this system were identified in the areas of tail injury prevention, nasal health, and the management of indoor housing and feeding. The performance of the top 20% of farms for each welfare indicator was used to establish benchmarks of: 0% to 5% clinical lameness, 0% to 12% of cows outside recommended BCS, 0% to 27% ocular discharge, 2% to 16% nasal discharge, 0% tail lacerations and docked tails, 0% to 3% tail breaks, 0% to 14% integument alterations, and 4% to 74% for avoidance distance of >1 m. These represent attainable targets for spring-calving pasture-based farms to promote good dairy cow welfare.

Body condition loss during the dry period: Insights from feeding behavior studies

Body condition change during the dry period (ΔBCS) has been associated with a myriad of transition cow diseases. We used data from 3 studies to assess the relationship between ΔBCS, feeding behavior, and body condition score (BCS) at dry-off. We also studied the mediation effect that dry matter intake (DMI) has on ΔBCS and the association between dry period feeding time and DMI. A total of 100 parous cows were enrolled in 3 studies to investigate differences in dry period diet on behavior, health, and performance pre- and postcalving. Body weight was measured and BCS was assessed by the same trained observer after dry-off and 1 wk from calving date. The ΔBCS was calculated by subtracting the BCS at calving minus the BCS at dry-off. The BCS at dry-off was categorized as overconditioned (≥3.5) or not overconditioned (<3.5); no cows had a BCS <2.75. Feeding behavior data were collected using electronic feed bins. Parity at dry-off (median = 2; min = 1, max = 6) and 305-d milk production (mean = 10,235 kg, SD = 1,625 kg) from the previous lactation were considered. Data sets were split into 2 time periods: d -56 to -22 (early) and -21 to 0 (late) in relation to calving. Selected feeding behaviors (DMI, DMI as a percentage of body weight, and feeding time) were used to evaluate the associations between each feeding behavior and BCS at dry-off in each period using mixed linear regression models. Each model included the following covariates: parity, previous 305-d milk yield, and trial treatment. Experimental day was included as random slope, and cow was included as random intercept. A mediation analysis was used to evaluate the potential causal direct effect of BCS at dry-off on ΔBCS and the potential indirect effect mediated by differences in DMI. The BCS at dry-off was associated with changes in feeding behavior, such that overconditioned cows had lesser daily DMI and feeding time during the early and late dry periods compared with not overconditioned animals. We also noted an effect of previous 305-d milk yield on DMI; cows that produced more milk had greater DMI throughout the dry period. The ΔBCS was only partially mediated by DMI, and BCS at dry-off still had a direct effect on ΔBCS. This result indicated that mechanisms other than DMI were associated with BCS loss during the dry period. Feeding time correlated weakly and moderately with DMI during the early and late dry periods, respectively. To conclude, strategies to improve intake during the dry period should take dry-off BCS into account or, preferably, efforts should be made to minimize the number of overconditioned cows at the end of lactation.

All-trans retinoic acid controls differentiation, proliferation, and lipolysis in isolated subcutaneous adipocytes from peripartal Holstein cows

Preadipocyte proliferation and differentiation are critical for normal adipose tissue development, including achieving a mature phenotype, characterized by its ability to accumulate triacylglycerol and release fatty acids. In nonruminants, it is well known that all-trans retinoic acid (ATRA), the most-active form of vitamin A, helps regulate proliferation, differentiation, and apoptosis in several types of cells including adipocytes. The purpose of this study was to evaluate the effect of ATRA on proliferation, apoptosis, differentiation, and lipolysis of primary bovine adipocytes isolated from subcutaneous adipose tissue of 5 healthy Holstein cows at 17 (±4 standard deviations) d postpartum. Cells were stimulated with increasing concentrations of ATRA (0.2, 2, and 20 nM) at the preconfluent (2 d) and postconfluent (8 d) preadipocyte stage or at the mature adipocyte stage (2 d). All concentrations of ATRA inhibited preconfluent preadipocyte proliferation with decreased proportion of S-phase cells and reduced protein abundance of cyclins (CCND1, CCND2, CCND3, CCNE1) and cyclin-dependent kinases (CDK2, CDK4, CDK6). Compared with vehicle, ATRA treatment induced apoptosis in preconfluent preadipocytes. Additionally, ATRA (0.2, 2, and 20 nM) supplementation also inhibited differentiation of postconfluent preadipocytes through downregulation of protein abundance of PPARγ and C/EBPα. After induction of differentiation, basal lipolysis in mature adipocytes increased upon treatment with all concentrations of ATRA. However, data on phosphorylated hormone-sensitive lipase or PLIN1 indicated that ATRA had no effect on epinephrine-stimulated lipolysis in mature adipocytes. Overall, these results demonstrate that ATRA might inhibit lipid accumulation by suppressing preadipocyte proliferation and differentiation, subsequently leading to apoptosis in postconfluent preadipocytes and promoting basal lipolysis in mature adipocytes. Overall, these in vitro responses provide some insights into the potential for nutritional management to modulate adipose tissue lipolysis, particularly in overconditioned cows during the dry period, which are more susceptible to suffer metabolic disorders due to excessive fat mobilization postpartum.

Validation of an Automated Body Condition Scoring System Using 3D Imaging

Body condition scores (BCS) measure a cow’s fat reserves and is important for management and research. Manual BCS assessment is subjective, time-consuming, and requires trained personnel. The BodyMat F (BMF, Ingenera SA, Cureglia, Switzerland) is an automated body condition scoring system using a 3D sensor to estimate BCS. This study assesses the BMF. One hundred and three Holstein Friesian cows were assessed by the BMF and two assessors throughout a lactation. The BMF output is in the 0–5 scale commonly used in France. We develop and report the first equation to convert these scores to the 1–5 scale used by the assessors in Ireland in this study ((0–5 scale × 0.38) + 1.67 → 1–5 scale). Inter-assessor agreement as measured by Lin’s concordance of correlation was 0.67. BMF agreement with the mean of the two assessors was the same as between assessors (0.67). However, agreement was lower for extreme values, particularly in over-conditioned cows where the BMF underestimated BCS relative to the mean of the two human observers. The BMF outperformed human assessors in terms of reproducibility and thus is likely to be especially useful in research contexts. This is the second independent validation of a commercially marketed body condition scoring system as far as the authors are aware. Comparing the results here with the published evaluation of the other system, we conclude that the BMF performed as well or better.

Metabolome profiling in skeletal muscle to characterize metabolic alterations in over-conditioned cows during the periparturient period

The transition from late gestation to early lactation is associated with extensive changes in metabolic, endocrine, and immune functions in dairy cows. Skeletal muscle plays an important role in maintaining the homeorhetic adaptation to the metabolic needs of lactation. The objective of this study was to characterize the skeletal muscle metabolome in the context of the metabolic changes that occur during the transition period in dairy cows with high (HBCS) versus normal body condition (NBCS). Fifteen weeks antepartum, 38 pregnant multiparous Holstein cows were assigned to 1 of 2 groups, which were fed differently to reach the targeted BCS and back fat thickness (BFT) until dry-off at -49 d before calving (HBCS: >3.75 and >1.4 cm; NBCS: <3.5 and <1.2 cm). During the dry period and the subsequent lactation, both groups were fed identical diets. The differences in both BCS and BFT were maintained throughout the study. The metabolome was characterized in skeletal muscle samples (semitendinosus muscle) collected on d -49, 3, 21, and 84 relative to calving using a targeted metabolomics approach (AbsoluteIDQ p180 kit; Biocrates Life Sciences AG, Innsbruck, Austria), which allowed for the quantification of up to 188 metabolites from 6 different compound classes (acylcarnitines, amino acids, biogenic amines, glycerophospholipids, sphingolipids, and hexoses). On d -49, the concentrations of citrulline and hydroxytetradecadienyl-l-carnitine in muscle were higher in HBCS cows than in NBCS cows, but those of carnosine were lower. Over-conditioning did not affect the muscle concentrations of any of the metabolites on d 3. On d 21, the concentrations of phenylethylamine and linoleylcarnitine in muscle were lower in HBCS cows than in NBCS cows, and the opposite was true for lysophosphatidylcholine acyl C20:4. On d 84, the significantly changed metabolites were mainly long-chain (>C32) acyl-alkyl phosphatidylcholine and di-acyl phosphatidylcholine, along with 3 long-chain (>C16) sphingomyelin that were all lower in HBCS cows than in NBCS cows. These data contribute to a better understanding of the metabolic adaptation in skeletal muscle of dairy cows during the transition period, although the physiological significance and underlying molecular mechanisms responsible for the regulation of citrulline, hydroxytetradecadienyl-l-carnitine, carnosine, and phenylethylamine associated with over-conditioning are still elusive and warrant further investigation. The changes observed in muscle lysophosphatidylcholine and phosphatidylcholine concentrations may point to an alteration in phosphatidylcholine metabolism, probably resulting in an increase in membrane stiffness, which may lead to abnormalities in insulin signaling in the muscle of over-conditioned cows.

Branched-chain amino acids: Abundance of their transporters and metabolizing enzymes in adipose tissue, skeletal muscle, and liver of dairy cows at high or normal body condition

Branched-chain amino acids (BCAA) are major components of milk protein and important precursors for nonessential AA. Thus, the BCAA transport and break-down play a key role in the metabolic adaptation to the high nutrient demands in lactation. However, in monogastrics, increased BCAA levels have been linked with obesity and certain metabolic disorders such as impaired insulin sensitivity. Our objective was to study the effect of over-conditioning at calving on plasma BCAA levels as well as the tissue abundance of the most relevant BCAA transporters and degrading enzymes in dairy cows during late pregnancy and early lactation. Thirty-eight Holstein cows were allocated 15 wk antepartum to either a normal- (NBCS) or over-conditioned (HBCS) group, receiving 6.8 or 7.2 MJ of NEL/kg of DM, respectively, during late lactation to reach the targeted differences in body condition score (BCS) and back fat thickness (BFT; NBCS: BCS <3.5, BFT <1.2 cm; HBCS: BCS >3.75, BFT >1.4 cm) until dry-off. During the dry period and next lactation, cows were fed the same diets, whereby differences in BCS and BFT were maintained: prepartum means were 3.16 ± 0.06 and 1.03 ± 0.07 cm (NBCS) vs. 3.77 ± 0.08 and 1.89 ± 0.11 cm (HBCS), postpartum means were 2.89 ± 0.06 and 0.81 ± 0.05 cm (NBCS) vs. 3.30 ± 0.06 and 1.38 ± 0.08 cm (HBCS). Blood and biopsies from liver, semitendinosus muscle, and subcutaneous adipose tissue (scAT) were sampled at d 49 antepartum, 3, 21, and 84 postpartum. Free BCAA were analyzed and the mRNA abundance of solute carrier family 1 member 5 (SLC1A5), SLC7A5, and SLC38A2 as well as branched-chain aminotransferase 2 (BCAT2), branched-chain α-keto acid dehydrogenase E1α (BCKDHA), and branched-chain α-keto acid dehydrogenase E1β (BCKDHB) as well as the protein abundance of BCKDHA were assessed. Concentrations of all BCAA changed with time, most markedly in HBCS cows, with a nadir around calving. Apart from Ile, neither individual nor total BCAA differed between groups. The HBCS group had greater BCKDHA mRNA as well as higher prepartum BCKDHA protein abundance in scAT than NBCS cows, pointing to a greater oxidative capacity for the irreversible degradation of BCAA transamination products in scAT of over-conditioned cows. Prepartum hepatic BCKDHA protein abundance was lower in HBCS than in NBCS cows. In both groups, SLC1A5, SLC7A5, and BCAT2 mRNA were most abundant in scAT, whereas SLC38A2 was higher in scAT and muscle compared with liver, and BCKDHA and BCKDHB mRNA were greatest in liver and muscle, respectively. Our results indicate that scAT may be a major site of BCAA uptake and initial catabolism, with the former, however, being independent of BCS and time relative to calving in dairy cows.

Associations between body condition score at parturition and microRNA profile in colostrum of dairy cows as evaluated by paired mapping programs

MicroRNA (miRNA) are abundant in milk, and likely have regulatory activity involving lactation and immunity. The objective of this study was to determine the miRNA profile in colostrum of overconditioned cows compared with cows of more moderate body condition score (BCS) at calving. Multiparous cows with either high (≥4.0 on a scale of 1 to 5; n = 7) or moderate BCS (2.75 to 3.50; n = 9) in the week before parturition were selected from a commercial dairy herd. Blood and colostrum were sampled within 24 h after calving. Blood serum was analyzed for free fatty acid (FFA) concentration. MicroRNA was isolated from colostrum samples after removing milk fat and cells. MicroRNA were sequenced, and reads were mapped to the bovine genome and to the existing database of miRNA at miRBase.org. Two programs, Oasis 2.0 and miRDeep2, were employed in parallel for read alignment, and analysis of miRNA count data was performed using DESeq2. Identification of differentially expressed miRNA from DESeq2 was not affected by the differences in miRNA detected by the 2 mapping programs. Most abundant miRNA included miR-30a, miR-148a, miR-181a, let-7f, miR-26a, miR-21, miR-22, and miR-92a. Large-scale shifts in miRNA profile were not observed; however, colostrum of cows with high BCS contained less miR-486, which has been linked with altered glucose metabolism. Colostrum from cows with elevated serum FFA contained less miR-885, which may be connected to hepatic function during the transition period. Potential functions of abundant miRNA suggest involvement in development and maintenance of cellular function in the mammary gland, with the additional possibility of influencing neonatal tissue and immune system development.

Metabolomics meets machine learning: Longitudinal metabolite profiling in serum of normal versus overconditioned cows and pathway analysis

This study aimed to investigate the differences in the metabolic profiles in serum of dairy cows that were normal or overconditioned when dried off for elucidating the pathophysiological reasons for the increased health disturbances commonly associated with overconditioning. Fifteen weeks antepartum, 38 multiparous Holstein cows were allocated to either a high body condition (HBCS; n = 19) group or a normal body condition (NBCS; n = 19) group and were fed different diets until dry-off to amplify the difference. The groups were also stratified for comparable milk yields (NBCS: 10,361 ± 302 kg; HBCS: 10,315 ± 437 kg; mean ± standard deviation). At dry-off, the cows in the NBCS group (parity: 2.42 ± 1.84; body weight: 665 ± 64 kg) had a body condition score (BCS) <3.5 and backfat thickness (BFT) <1.2 cm, whereas the HBCS cows (parity: 3.37 ± 1.67; body weight: 720 ± 57 kg) had BCS >3.75 and BFT >1.4 cm. During the dry period and the subsequent lactation, both groups were fed identical diets but maintained the BCS and BFT differences. A targeted metabolomics (AbsoluteIDQ p180 kit, Biocrates Life Sciences AG, Innsbruck, Austria) approach was performed in serum samples collected on d -49, +3, +21, and +84 relative to calving for identifying and quantifying up to 188 metabolites from 6 different compound classes (acylcarnitines, AA, biogenic amines, glycerophospholipids, sphingolipids, and hexoses). The concentrations of 170 metabolites were above the limit of detection and could thus be used in this study. We used various machine learning (ML) algorithms (e.g., sequential minimal optimization, random forest, alternating decision tree, and naïve Bayes-updatable) to analyze the metabolome data sets. The performance of each algorithm was evaluated by a leave-one-out cross-validation method. The accuracy of classification by the ML algorithms was lowest on d 3 compared with the other time points. Various ML methods (partial least squares discriminant analysis, random forest, information gain ranking) were then performed to identify those metabolites that were contributing most significantly to discriminating the groups. On d 21 after parturition, 12 metabolites (acetylcarnitine, hexadecanoyl-carnitine, hydroxyhexadecenoyl-carnitine, octadecanoyl-carnitine, octadecenoyl-carnitine, hydroxybutyryl-carnitine, glycine, leucine, phosphatidylcholine-diacyl-C40:3, trans-4-hydroxyproline, carnosine, and creatinine) were identified in this way. Pathway enrichment analysis showed that branched-chain AA degradation (before calving) and mitochondrial β-oxidation of long-chain fatty acids along with fatty acid metabolism, purine metabolism, and alanine metabolism (after calving) were significantly enriched in HBCS compared with NBCS cows. Our results deepen the insights into the phenotype related to overconditioning from the preceding lactation and the pathophysiological sequelae such as increased lipolysis and ketogenesis and decreased feed intake.

Level and source of fat in the diet of gestating beef cows: I. Effects on the prepartum performance of the dam and birth weight of the progeny1.

A 2-yr study was conducted to evaluate the effects of level and source of fat in the diet of gestating beef cows on their prepartum performance and birth weight of progeny. Each year, 75 multiparous (≥3 calving) pregnant Angus cows were stratified by BW (663 ± 21.5 kg) and BCS (2.6 ± 0.12; 1 to 5 scale) and randomly assigned to 1 of 15 outdoor pens. Subsequently, each pen was randomly assigned to 1 of 3 (n = 5) treatments: a low-fat diet (LF; 1.4 ± 0.12% EE) consisting of grass-legume hay, barley straw, and barley grain, or 1 of 2 high-fat diets (HF; 3.3 ± 0.20% EE) that included either a canola seed (CAN) or a flaxseed (FLX) based pelleted feed. Diets were formulated to meet the requirements of pregnant beef cows during the last 2 trimesters of gestation (0.183 ± 4.8 d), adjusted for changes in environmental conditions, and offered such that each pen on average received similar daily amounts of DE (31.2 ± 2.8 Mcal/cow), CP (1.36 ± 0.13 kg/cow), and DM (12.9 ± 1.0 kg/cow). Data were analyzed as a randomized complete block design with contrasts to separate the effects of level (LF vs. HF) and source (CAN vs. FLX) of fat. After 160 d on trial, conceptus corrected-BW (CC-BW) of LF cows (708 kg) and the proportion of overconditioned cows (13.2%) were greater (P ≤ 0.04) than those of HF, with no difference (P ≥ 0.84) between CAN and FLX for CC-BW (697 kg) and proportion of overconditioned cows (3.6% vs. 2.9%). Feeding FLX diet during gestation resulted in cows with a greater (P ≤ 0.01) concentration of conjugated linolenic acid (0.12% vs. 0.05%) and n-3 (0.58% vs. 0.37%) fatty acids, and a tendency (P = 0.09) for conjugated linoleic acid concentration (1.05% vs. 0.88%) to be greater in subcutaneous adipose tissue (SCAT) when compared with cows fed the CAN diet. By the end of gestation, serum NEFA concentration of LF cows (592 µEq/L) was lower (P < 0.01) than that of HF cows, and FLX cows had greater (P < 0.01) serum NEFA concentration than CAN cows (636 vs. 961 µEq/L). Cows receiving the LF diet during gestation gave birth to lighter (P < 0.01) calves compared with those receiving the HF diets (40.2 vs. 42.9 kg), with no difference (P = 0.24) between calves born to CAN (42.4 kg) and FLX (43.3 kg) cows. In conclusion, these results suggest a partitioning of the ME in pregnant beef cows that is dependent on the type of dietary energy, resulting in heavier calves at birth for cows fed high-fat diets. Also, the type of fatty acid in the diet of gestating beef cows affected the fatty acid profile in SCAT and serum NEFA concentration.

Prepartum body conditions affect insulin signaling pathways in postpartum adipose tissues in transition dairy cows

BackgroundOverconditioned dairy cows are susceptible to excessive lipolysis and increased insulin resistance during the transition period. The associations among body fat reserve, insulin resistance, and lipolysis in adipose tissues (AT) remain to be elucidated. Therefore, this study aimed to investigate whether excessive fat reserves influence the insulin signaling pathway in AT postpartum.ResultsTwenty multiparous dairy cows were selected and assigned to one of two groups, according to prepartum body condition score (BCS): Control group (BCS = 3.0–3.5; n = 10) and Overconditioned group (BCS ≥ 4.0; n = 10). Blood samples were collected on days −14, −7, −4, −2, −1, 0, 1, 2, 4, 7, and 14 relative to parturition. Subcutaneous AT were collected on day 2 following parturition for quantitative real-time polymerase chain reaction and western blot analyses. No differences were observed between the two groups in serum glucose, non-esterified fatty acids, β-hydroxybutyric acid, tumor necrosis factor (TNF) α, insulin, or leptin concentrations during the experimental period. Compared with the control cows, the overconditioned cows had lower serum triglyceride levels and higher adiponectin concentrations. In the AT postpartum, insulin receptor mRNA and protein levels were lower in the overconditioned cows than in the control cows, and no differences were found in glucose transporter 4 mRNA. Compared with the control cows, the overconditioned cows had lower mRNA levels of TNFα and higher mRNA levels of peroxisome proliferator-activated receptor gamma (PPARγ) in AT postpartum. The phosphorylated protein kinase B (AKT) content and phosphorylation rate of AKT were increased in the overconditioned cows compared with the control cows, which suggested that the downstream insulin signaling in AT was affected.ConclusionsIn the present study, transition dairy cows with higher BCS did not show more fat mobilization. The changes of insulin signaling pathway in AT postpartum of overconditioned cows may be partly related to the expression of PPARγ and TNFα, and the secretion of adiponectin.

PSVII-32 Profiling peripheral microRNA in normal- versus over-conditioned dairy cows during dry-off and early lactation.

PSVII-32 Profiling peripheral microRNA in normal- versus over-conditioned dairy cows during dry-off and early lactation.

Effect of injectable trace mineral complex supplementation on development of ovarian structures and serum copper and zinc concentrations in over-conditioned Holstein cows

This study evaluated the effect of injecting trace minerals on reproductive performance in over-conditioned Holstein cows before synchronized estrus. Multiparous non-lactating, over-conditioned repeat breeder cows (n=20) were assigned randomly to one of two treatments: 1) control (n=10), and 2) supplementation with an injectable trace mineral complex 25days before expected synchronized estrus (n=10). Follicular waves were synchronized by intravaginal insertion of a CIDR for eight days and an intramuscular (i.m.) injection of a GnRH analogue. Estrus was induced at CIDR removal by an i.m. injection of PGF2α. Blood samples were collected before and after synchronized estrus. The response variables were follicle population (FP), diameter of the preovulatory follicle at CIDR removal (DFP0) and at estrus detection (DFP1), time of estrus after CIDR removal (TE), area of corpus luteum (ACL), pregnancy rate and copper and zinc serum concentrations. The statistical analysis of the variables was carried out with SAS. The FP, DFP0, DFP1, TE, ACL and serum concentrations of copper and zinc were not affected by the trace mineral injection (P>0.05). Even though pregnancy rate at 40 (77.78±13.46 vs 44.44±16.56%) and 60days after AI (66.67±15.71 vs 33.33±15.71%) was numerically higher for cows injected with trace minerals than for the control group, the differences were not significant (P>0.05). In conclusion, while follicular and corpus luteum development were not affected by trace mineral injection, it may be a feasible way to increase the pregnancy rate in over-conditioned cows.

Ultrastructure of Cell Organelles in Pre-implantation Embryos from Cows with Different Body Condition Score.

Morphology of important cell organelles (mitochondria, lipid droplets, vacuoles, inclusion bodies and apoptotic bodies) in embryos derived from cows with different body condition score (BCS) was analysed by transmission electron microscopy (TEM). Embryos were recovered on 7th day after the insemination by a standard non-surgical flushing of the uterine horns from superovulated Holstein Friesian cows with BCS 2, 3, 4 and 5. Thereafter, the good quality blastocysts were processed for TEM. The electronograms were evaluated by stereological analysis. The relative volume of lipid droplets in BCS4 and BCS5 embryos increased significantly (18.53 and 22.40%) when compared to BCS3 embryos (5.46%). In the embryos from the BCS4 or BCS5 cows, we observed different morphological patterns of mitochondria, as well as the mitochondria containing vacuoles. BCS4 and BCS5 embryo cell nuclei showed the structure typical for low transcription activity (none or very few reticular nucleoli); also dilated inter-cellular spaces were often observed in these embryos. In conclusion, differences in the ultrastructural morphology of embryos from over-conditioned cows (BCS4 and BCS5), particularly the higher lipid content in the cytoplasm, can be a marker of their low quality, and this fact can be a contributing factor to subfertility in over-conditioned cows.

Short Communication: Immunohistochemical localization of the immune cell marker CD68 in bovine adipose tissue: impact of tissue alterations and excessive fat accumulation in dairy cows

With the onset of lactation energy from feed intake is mostly insufficient to meet the requirements of dairy cows. Lipid mobilization from adipose tissue (AT) could lead to a compromised inflammatory response enhancing the incidence for diseases. In addition, tissue alterations can occur, displaying areas of necrosis and inflammation. Furthermore, over-conditioned cows mobilizing more lipids from AT than thin cows are prone to develop metabolic disorders. This might lead to an increased infiltration of phagocytic immune cells into AT. In the present study, CD68 positive cells were localized in AT from 10 early lactating Holstein cows displaying different grades of AT alterations. Biopsies were sampled from visceral and subcutaneous AT and the number of CD68 positive cells was immunohistochemically determined. In addition, AT biopsies from over-conditioned, non-pregnant, non-lactating cows (n=8) were immunohistochemically analyzed for CD68 positive cells. The percentage of CD68 positive cells was less than 2% in AT biopsies with tissue alterations and in AT from over-conditioned cows. Therefore, immune cell infiltration demonstrated via the localization of CD68 positive cells seems to play only a minor role in AT from over-conditioned cows as well as in different bovine AT depots with tissue alterations.