Subclinical Ketosis Research Articles

Investigation of Ketosis Prevalence in Dairy Cows in Bingöl Province of Turkey

The aim of this study is to investigate the prevalence of ketosis in 7 different villages (Alibir, Ardıçtepe, Çavuşlar, Garip, Sudüğünü, Yamaç, and Yeşil) of Bingöl province. A total of 411 dairy cows, including Simmental, Holstein and crossbred breeds, were used in the study during the 3-week period after birth. Ketosis was diagnosed by measuring blood beta-hydroxy-butyric acid (BHBA) concentration. Blood BHBA concentrations were analyzed using the FreeStyle Optium Neo H (Abbott Diabetes Care Ltd., Witney, UK). Blood BHBA ≥2.6 mmol/L was determined as clinical ketosis (CK), BHBA≥1.2 mmol/L as subclinical ketosis (SCK), and BHBA

Elevated RBP4 in Subclinical Ketosis Cows Inhibits Follicular Granulosa Cell Proliferation and Steroid Hormone Synthesis.

The mechanism by which subclinical ketosis (SCK) causes postpartum reproductive disorders in dairy cows remains unclear. In this study, cows within the day 14 to 21 postpartum period were categorized into the SCK group or the control group. Subsequently, they were monitored until 45 d to 60 d postpartum and divided into the SCK anestrus group (SCK-AE, n = 12) and the control estrus group (C-E, n = 12). In comparison to the C-E group, the RBP4 and p-AKT of the SCK-AE group exhibited increased levels in serum, liver, and ovaries. In the in vitro experimental cultivation of granulosa cells (GCs), after adding RBP4, cell proliferation, steroid hormone secretion and synthesis, and GLUT4 secretion were inhibited, and cell apoptosis was exacerbated. After silencing STRA6 (RBP4 receptor), cell proliferation and steroid hormone secretion and synthesis, as well as the inhibition of GLUT4, were alleviated, and the situation of cell apoptosis also improved. The SC79 activator could promote the phosphorylation of AKT, thus alleviating the increased cell proliferation, steroid hormone secretion and synthesis, GLUT4 inhibition, and apoptosis rate in cow GCs induced by RBP4 stimulation. Our research indicates that elevated RBP4 levels in SCK cows inhibit the proliferation, apoptosis, and steroid hormone synthesis of GCs through the STRA6 receptor and the PI3K/AKT pathway.

Short-term reduction in feed intake by dairy cows in the postpartum period leads to subclinical ketosis development

Subclinical ketosis is widespread in highly productive dairy cows after calving and often remains undiagnosed, leading to reduced productivity. Physiologically controlled feeding in the first weeks after calving and during the intensive lactation period can reduce the incidence of ketosis. The study aimed to determine how a short-term reduction of feed after calving affects the formation of ketone bodies in blood, urine and milk of dairy cows. The group of ten Ukrainian black-spotted dairy breed cows after calving aged from 4 to 6 years was involved in the experiment lasted for 72 h. In 24 h the amount of compound feed, haylage, and silage received by cows was gradually reduced until complete exclusion in the diet. Blood, urine, and milk samples were collected three times a day. The content of ketone bodies, β-hydroxybutyrate and glucose was estimated with the corresponding sets of indicator strips. The content of total bilirubin, cholesterol, albumin and enzymes activity in the blood serum were determined on biochemical analyzer. It was found that in 24 h after the beginning of feed reduction the level of blood glucose decreased, persistent hypoglycemia within 48 and 72 h was developed, the concentration of β-hydroxybutyrate in blood and milk and of ketone bodies in urine was elevated. The increase in total bilirubin concentration and liver enzymes activity in the blood serum with a simultaneous decrease in albumin level and delayed sodium propionate conversion into glucose were observed. Thus, short-term reduction in feed intake by dairy cows after calving causes ketosis development and violation of liver functions. Keywords: dairy сows, feeding level, glucose, ketosis, liver damage, β-hydroxybutyrate

Risk Factors and metabolic indicators associated with the percentage frequency of subclinical Ketosis on Ouled Djellal Ewes in Eastern Highlands of Algeria

Risk Factors and metabolic indicators associated with the percentage frequency of subclinical Ketosis on Ouled Djellal Ewes in Eastern Highlands of Algeria

Effect of Replacing Corn Grain and Soybean Meal with Field Peas at Different Levels on Feed Intake, Milk Production, and Metabolism in Dairy Cows under a Restrictive Grazing.

This study assessed the effects of replacing soybean meal (SBM) and corn grain with field peas in the concentrate of grazing dairy cows on milk production, intake, ruminal fermentation, and blood indicators. Twelve multiparous lactating Holstein-Friesian cows were utilized in a replicated 3 × 3 Latin square design, comprising three periods and three treatments: (1) Pea-0 (Control diet): 6 kg dry matter (DM) of fresh pasture, 7.2 kg DM of grass silage, and 7 kg DM of a concentrate containing 0% pea; (2) Pea-30: Control diet with the concentrate composed of 30% pea; (3) Pea-60: Control diet with the concentrate composed of 60% pea. The effect of treatments on productive and metabolic parameters was evaluated using linear-mixed models. Pasture and total DM intake, milk production, and composition were unaffected by treatments. Despite the concentrates being isonitrogenous and isoenergetic, crude protein (CP) intake was slightly higher in Pea-30 and significantly higher in Pea-60 due to higher pasture CP content in the pasture grazed by these groups, leading to higher milk urea content, though within recommended ranges. Blood parameters showed no significant changes, except for plasma β-hydroxybutyrate, which was lowest in the Pea-60 treatment; however, all values were within ranges not indicative of subclinical ketosis. Ruminal fermentation parameters were similar across treatments. These findings support the use of field peas as a viable alternative to replace SBM and corn grain in concentrates, enabling similar milk production and composition in grazing dairy cows.

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.

Functionalized Graphene-Based Biosensors for Early Detection of Subclinical Ketosis in Dairy Cows

Precision livestock farming is one method of improving the productivity, health, and well-being of animals. Advanced diagnostic tools including biosensors can play a key role in the management of livestock operations. Detection of ketosis, a metabolic disease that occurs in early lactation dairy cows due to the negative energy balance, is one potential on-farm use of biosensors. Beta–hydroxybutyrate (βHB) is an excellent biomarker for ketosis monitoring in dairy cows because βHB is one of the main ketones produced during this metabolic state. In this report, we develop a low-cost, Keto-sensor (graphene-based sensor) for the detection and quantification of βHB concentrations in less than a minute. In this device, graphene nanosheets were layered onto a screen–printed electrode (SPE), and then a stabilized enzyme (Beta–hydroxybutyrate dehydrogenase, NADH, and glycerol) was used to functionalize the graphene surface enabled by EDC–NHS conjugation chemistry. The Keto-sensor offers an analytical sensitivity of 10 nM and a limit-of-detection (LoD) of 0.24 nM within a detection range of 0.00001-3.0 mM. Spike testing indicates that the Keto-sensor can detect βHB in serum samples from bovines with subclinical ketosis. The Keto-sensor developed in this study shows promising results for early detection of subclinical ketosis on farms.

The Effect of Subclinical Ketosis on the Peripheral Blood Mononuclear Cell Inflammatory Response and Its Crosstalk with Depot-Specific Preadipocyte Function in Dairy Cows.

During the periparturient period, cows undergo heightened energy demands at lactation onset, paired with reduced dry matter intake, leading to negative energy balance (NEB). Excessive lipolysis-driven adipose tissue remodeling, triggered by NEB, significantly contributes to ketosis in periparturient dairy cows. However, the role of peripheral blood mononuclear cells (PBMCs) in the pathogenesis of ketosis and in modulating adipose tissue function remains poorly understood. Here, we investigated how ketosis affects the transcriptional profile and secretome of PBMCs and its influence on preadipocyte function in visceral adipose tissue (VAT) and subcutaneous adipose tissue (SAT). Twenty-one postpartum Holstein dairy cows were categorized as either subclinical ketosis (SCK; BHB ≥ 1.0 mM) or control (CON; BHB < 0.8 mM) based on blood beta-hydroxybutyrate (BHB) concentration screening. Blood samples were collected intravenously for the isolation of PBMCs and serum metabolic profiling. Ketosis elevated circulating NEFA and BHB levels but reduced total WBC and neutrophil counts. Isolated PBMCs were evaluated for gene expression and used to produce conditioned media (PBMC-CM), during which PBMCs were stimulated with 10 ng/mL LPS. The overall phenotype of PBMCs was largely consistent between SCK and CON cows, with minimal differences detected in immunomodulatory cytokine expression and PBMC-CM composition following stimulation. Preadipocytes isolated from non-ketotic cows were treated with PBMC-CM to assess the effect of PBMC secretomes on adipose cell function. Preadipocytes treated with SCK PBMC-CM showed reduced lipid accumulation compared to those treated with CON PBMC-CM regardless of the depot. SAT preadipocytes had heightened expression of lipid metabolism-related genes, including DGAT1, LIPE, and FASN, compared to VAT when treated with SCK PBMC-CM. Preadipocytes treated with CM from PBMC stimulated by LPS exhibited upregulation in IL1B and IL6 regardless of the depot or source of PBMCs. Together, these results indicate that although PBMC profiles showed minimal differences, preadipocytes treated with PBMC-CM may be influenced by additional factors, leading to altered preadipocyte function and gene expression that may contribute to adipose cellular dysfunction.

Metabolic adaptation to lactation of dairy cows in two contrasting facilities involving partial confinement plus grazing or total confinement

Context The increasing intensification of pasture-based systems has subjected the cows to different productive environments, which could affect physiological responses and, consequently, their productive performance. Aims The aim was to determine the effect of contrasting housing facilities (outdoor soil-bedded vs compost-bedded pack barn) used in partial confinement in pasture-based systems, on the metabolic adaptation during early lactation and its impact on productive and reproductive performance of autumn- and spring-calving dairy cows, having as a positive control a zero-grazing confined system in compost barn. Methods Multiparous Holstein dairy cows that calved in autumn (n = 36) and spring (n = 48) were distributed in the following three treatments from calving to 90 days in milk (DIM): outdoor soil-bedded–grazing (OD-GRZ), compost barn–grazing (CB-GRZ), compost barn–total mixed ration (CB-TMR). Milk production, total milksolids (TMS), body condition score, non-esterified fatty acids (NEFA), beta-hydroxybutyrate (BHB), cholesterol, insulin, insulin-like growth factor-1 (IGF-1), and the proportion of cows with corpus luteum were determined in repeated measurements. Key results In both calving seasons, milk production and TMS yields did not differ between OD-GRZ and CB-GRZ, but were greater in CB-TMR (P &lt; 0.0001). In autumn, NEFA and BHB were not affected by treatments, but cholesterol increased faster in CB-TMR (P = 0.0500). In spring, NEFA and cholesterol concentrations were not affected by treatment, but BHB remained greater in OD-GRZ until 90 DIM than in the other treatments. IGF-1 and insulin did not differ between treatments in autumn, but in spring, IGF-1 was greater in CB-TMR (P &lt; 0.0001). The proportion of cows with corpus luteum was not different between pasture-based treatments, but both were greater than CB-TMR up to 40 DIM in autumn (P = 0.0489) and during the entire study in spring CB-TMR (P = 0.0285). Conclusions Although no differences were found among housing facilities in partial confinement, except that in spring OD-GRZ cows had greater BHB concentrations, confined cows prioritised milk production instead of reproductive function, despite presenting better energy indicators than did pasture-based cows. Implications Outdoor housing combined with grazing can increase the risk of greater BHB concentrations, indicative of subclinical ketosis, under heat-stress conditions, when compared with indoor housing. Confined cows increased milk production but had a delay in the resumption of ovarian cyclicity, even having a better energy status than pasture-based cows.

Changes in adiponectin levels of subclinical ketosis cows and their effects on steroid hormone secretion and proliferation in follicular granulosa cells

In dairy cows, the occurrence of subclinical ketosis (SCK) is particularly high during early lactation. Previously, we documented alterations in the abundance of adiponectin (ADPN) in anestrus cows with SCK in comparison to cows in estrus. In the present study, 60 cows were divided into two groups: control (C, n = 30) and SCK (n = 30). Based on cow’s estrus situation in two group at 55–60 days postpartum, 15 anestrus SCK cows and estrus cows were designated the SCK-A group and C-E group, respectively. The SCK-A group had downregulated serum and follicular fluid ADPN levels compared with the C-E group. The serum ADPN level was positively correlated with the insulin level and follicle growth rate, and there was a positive correlation between ADPN and glucose in the follicular fluid. Primary culture of dairy cow granulosa cells (GCs) was established to observe the effect of low glucose (Glu) and/or ADPN on GCs cyclins and proteins important for steroid synthesis. The results showed that the addition of 1 µg/mL ADPN alleviated the negative effects of low Glu treatment on the proliferation of GCs and the expression of steroid secretion related protein proteins. Treatment with LY294002 (PI3K inhibitor) four experimental GCs groups: control (0 µg/mL ADPN), 1 µg/mL ADPN, LY294002 inhibitor, and 1 µg/mL ADPN+LY294002. The results showed that ADPN promotes the secretion of steroid hormones by GCs through the PI3K–AKT. In summary, ADPN plays a crucial role in ameliorating postpartum anestrus in dairy cows with SCK.

Global losses due to dairy cattle diseases: A comorbidity-adjusted economic analysis

An economic simulation was carried out over 183 milk-producing countries to estimate the global economic impacts of 12 dairy cattle diseases and health conditions: mastitis (subclinical and clinical), lameness, paratuberculosis (Johne's disease), displaced abomasum, dystocia, metritis, milk fever, ovarian cysts, retained placenta, and ketosis (subclinical and clinical). Estimates of disease impacts on milk yield, fertility, and culling were collected from the literature, standardized, meta-analyzed using a variety of methods ranging from simple averaging to random-effects models, and adjusted for comorbidities to prevent overestimation. These comorbidity-adjusted disease impacts were then combined with a set of country-level estimates for lactational incidence or prevalence or both, herd characteristics, and price estimates within a series of Monte Carlo simulations that estimated and valued the economic losses due to these diseases. It was estimated that total annual global losses are US$65 billion (B). Subclinical ketosis, clinical mastitis, and subclinical mastitis were the costliest diseases modeled, resulting in mean annual global losses of approximately US$18B, US$13B, and US$9B, respectively. Estimated global annual losses due to clinical ketosis, displaced abomasum, dystocia, lameness, metritis, milk fever, ovarian cysts, paratuberculosis, and retained placenta were estimated to be US$0.2B, US$0.6B, US$0.6B, US$6B, US$5B, US$0.6B, US$4B, US$4B, and US$3B, respectively. Without adjustment for comorbidities, when statistical associations between diseases were disregarded, mean aggregate global losses would have been overestimated by 45%. Although annual losses were greatest in India (US$12B), the United States (US$8B), and China (US$5B), depending on the measure of losses used (losses as a percentage of gross domestic product, losses per capita, losses as a percentage of gross milk revenue), the relative economic burden of these dairy cattle diseases across countries varied markedly.

Application of an electronic nose for the diagnosis of ketosis in dairy cows

Ketosis in dairy cows is a nutritional and metabolic disease that severely affects the milk industry and dairy cow health. An electronic nose can identify volatile organic compounds (VOCs). This study applied the electronic nose to detect VOCs in dairy cow milk, feces, and blood. Blood ketone bodies (BHBA) were measured, and 10 ketosis cows, 71 subclinical ketosis cows, and 75 healthy cows were considered the research subjects. Cow milk, feces, and blood samples were collected for electronic nose testing. The measured VOC data were processed through multivariate analysis and machine learning algorithms. The electronic nose analysis results were validated through the GC-MS analysis of the milk samples. The results unveiled spatial differences in the electronic nose VOC values of milk, feces, and blood, and the cumulative contribution rates of both principal component variables (PC) 1 and PC2 reached more than 95%. The machine learning algorithm results revealed that the accuracy of the milk probabilistic neural network (PNN) was 87.5%, the stool support vector machine was 100%, and the blood PNN was 66.7%. This result revealed that electronic nose detection can effectively differentiate healthy, subclinical ketosis, and clinical ketosis cows. According to the milk GC-MS results, significant differences were noted in the VOC content of milk samples between the different groups.The aforementioned studies revealed that electronic nose can aid in the effective prediction and diagnosis of ketosis in dairy cows. This lays the foundation for establishing a new method for diagnosing ketosis in dairy cows.

Multi-Omics Reveals Disrupted Immunometabolic Homeostasis and Oxidative Stress in Adipose Tissue of Dairy Cows with Subclinical Ketosis: A Sphingolipid-Centric Perspective.

Ketosis, especially its subclinical form, is frequently observed in high-yielding dairy cows and is linked to various diseases during the transition period. Although adipose tissue plays a significant role in the development of metabolic disorders, its exact impact on the emergence of subclinical ketosis (SCK) is still poorly understood. The objectives of this study were to characterize and compare the profiling of transcriptome and lipidome of blood and adipose tissue between SCK and healthy cows and investigate the potential correlation between metabolic disorders and lipid metabolism. We obtained blood and adipose tissue samples from healthy cows (CON, n = 8, β-hydroxybutyric acid concentration < 1.2 mmol/L) and subclinical ketotic cows (SCK, n = 8, β-hydroxybutyric acid concentration = 1.2-3.0 mmol/L) for analyzing biochemical parameters, transcriptome, and lipidome. We found that serum levels of nonesterified fatty acids, malonaldehyde, serum amyloid A protein, IL-1β, and IL-6 were higher in SCK cows than in CON cows. Levels of adiponectin and total antioxidant capacity were higher in serum and adipose tissue from SCK cows than in CON cows. The top enriched pathways in whole blood and adipose tissue were associated with immune and inflammatory responses and sphingolipid metabolism, respectively. The accumulation of ceramide and sphingomyelin in adipose tissue was paralleled by an increase in genes related to ceramide biosynthesis, lipolysis, and inflammation and a decrease in genes related to ceramide catabolism, lipogenesis, adiponectin production, and antioxidant enzyme systems. Increased ceramide concentrations in blood and adipose tissue correlated with reduced insulin sensitivity. The current results indicate that the lipid profile of blood and adipose tissue is altered with SCK and that certain ceramide species correlate with metabolic health. Our research suggests that disruptions in ceramide metabolism could be crucial in the progression of SCK, exacerbating conditions such as insulin resistance, increased lipolysis, inflammation, and oxidative stress, providing a potential biomarker of SCK and a novel target for nutritional manipulation and pharmacological therapy.

Hemato-biochemical and Ultrasonographic Studies on Subclinical Ketosis in Dairy Cows

Hemato-biochemical and Ultrasonographic Studies on Subclinical Ketosis in Dairy Cows

Identification of genomic regions and genes associated with subclinical ketosis in periparturient dairy cows.

Subclinical ketosis (SCK) is a prevalent metabolic disorder that occurs during the transition to lactation period. It is defined as a high blood concentration of ketone bodies (beta-hydroxybutyric acid f ≥ 1.2 mmol/L) within the first few weeks of lactation, and often presents without clinical signs. SCK is mainly caused by negative energy balance (NEB). The objective of this study is to identify single nucleotide polymorphisms (SNPs) associated with SCK using genome-wide association studies (GWAS), and to predict the biological functions of proximal genes using gene-set enrichment analysis (GSEA). Blood samples were collected from 112 Holstein cows between 5 and 18 days postpartum to determine the incidence of SCK. Genomic DNA extracted from both SCK and healthy cows was examined using the Illumina Bovine SNP50K BeadChip for genotyping. GWAS revealed 194 putative SNPs and 163 genes associated with those SNPs. Additionally, GSEA showed that the genes retrieved by Database for Annotation, Visualization, and Integrated Discovery (DAVID) belonged to calcium signaling, starch and sucrose, immune network, and metabolic pathways. Furthermore, the proximal genes were found to be related to germ cell and early embryo development. In summary, this study proposes several feasible SNPs and genes associated with SCK through GWAS and GSEA. These candidates can be utilized in selective breeding programs to reduce the genetic risk for SCK and subfertility in high-performance dairy cows.

Randomized clinical trial of ketoprofen or ceftiofur for treatment of metritis in dairy cows

Our objectives were to compare the efficacy of ketoprofen or ceftiofur for treatment of metritis in dairy cows considering subsequent health, production, and reproduction. Cows from 2 commercial dairy farms in Ontario, Canada were examined with a Metricheck device 3 times per week from 2 to 14 d in milk (DIM). Cows with metritis (fetid vaginal discharge; n = 193) were blocked by parity and fever (rectal temperature ≥39.5°C or <39.5°C) and within each block per farm, randomly assigned to receive 3 mg/kg BW of ketoprofen (KET) or 2.2 mg/kg of ceftiofur hydrochloride (CEF), once a day for 3 d. Day of enrollment was considered study d 0. Rectal temperature and attitude were evaluated in cows with metritis on study d 0, 3, 4, 7, 10, and 13, and vaginal discharge was evaluated on study d 4, 7, 10, and 13. Body condition was scored at enrollment and 35 DIM, and serum concentration of haptoglobin was measured at d 0, 2, 4, and 7. Cows with rectal temperature ≥39.5°C or a depressed attitude on d 3 were classified as clinical failure and received treatment with ceftiofur for 3 d (KET), or 2 additional days (CEF), to a maximum of 5 d of treatment with ceftiofur. At 35 ± 3 DIM cows were examined for uterine involution by transrectal palpation, purulent vaginal discharge (PVD) by Metricheck, and endometritis by endometrial cytology. Time to onset of cyclicity was assessed by serum progesterone (P4) measurements at 28, 42, and 56 DIM. Contemporary cows from the same farms without metritis (NOMET; n = 1,043) were used for comparison. Data were analyzed with mixed linear or logistic regression or Cox's proportional hazard models, including herd as a random effect. The proportion of clinical resolution of metritis on d 3 (96% vs. 92%), of cows with fever (from d 3 to d 13 after enrollment) or fetid discharge (from d 4 to d 13 after enrollment), and the number of medical treatments (3.1 vs. 3.3) were not different between CEF and KET, respectively. Cows in KET received fewer antibiotic treatments than cows in CEF (0.3 vs. 3.1). Uterine involution, the prevalence of PVD (50% vs. 47%) and subclinical endometritis (6.6% vs. 4.3%), and the proportion of cyclic cows (82% vs. 86%) did not differ between CEF and KET. Cows in KET had greater serum haptoglobin concentration from d 2 to 7 after enrollment. The incidence of mastitis, lameness, or displaced abomasum to 60 DIM and subclinical ketosis to 21 DIM did not differ among CEF, KET, and NOMET. There were no differences in median days to first AI (CEF = 68 d; 95% CI: 65-70; KET = 69 d; 95% CI: 68-72; NOMET = 69 d; 95% CI: 68-70), and median days to pregnancy (CEF = 118 d; 95% CI: 92-145; KET = 113 d; 95% CI: 90-135; NOMET = 105 d; 95% CI: 101-109), pregnancy at first AI at 33 d after insemination (CEF = 42%; KET = 41%; NOMET = 41%), pregnancy loss after first AI (CEF = 8%; KET = 11%; NOMET = 8%), hazard of pregnancy or hazard of culling up to 300 DIM. Milk yield was not different between CEF and KET during the first 10 weeks, but lesser in KET at wk 2 and 4 and CEF at wk 2, 4, and 6 than in NOMET. In this pilot-scale study, given early detection, we did not detect differences in subsequent health, milk yield, or reproductive performance in cows with metritis initially treated for 3 d with CEF or KET. Additional, larger studies are warranted.

Changes in blood biochemical parameters in highly productive cows with ketosis.

Biochemical blood testing is the main diagnostic indicator of the clinical condition of highly productive animals and a method of determining changes in metabolic disorders. This study focuses on metabolic changes (ketosis), which are of the utmost importance in the assessment of the health status of animals, as well as differences in intergroup characteristics. The main focus of this study is to demonstrate the influence of subclinical ketosis in highly productive cows on changes in biochemical blood parameters during different physiological periods to further prevent this disease, adjust feeding rations, and prevent premature culling of animals. This study aimed to evaluate and establish changes in the biochemical status dynamics of highly productive cows with metabolic disorders in an industrial livestock complex. Blood samples were systematically collected from highly productive cows of the Simmental breed (n = 60) and served as the primary material for subsequent analyses. Each methodological step was designed to ensure evaluation of the metabolic changes associated with post-calving adjustments in highly productive dairy cows. This study employed a comprehensive approach integrating clinical assessments, laboratory analyses, biochemical evaluations, instrumental measurements, and statistical analyses. A biochemical blood test showed that the number of ketone bodies in the experimental group exceeded the norm, varied depending on the physiological state of the animals, and ranged from 0.89 to 1.45 mmol/L. At 10 days after calving, the highest indicator was 1.45 ± 0.05 mmol/L. This indicator was 1.05 mmol/L higher than that in the control group and exceeded the norm by 0.95. Excess ketone bodies in the blood of animals led to accumulation in urine and milk, indicating a disturbance in metabolic processes in the body and a decrease in the quality of animal husbandry products. The sample size and the focus on a single breed from one geographical location may limit the generalizability of the findings. Further research should explore the mechanistic bases of ketosis development, potentially integrating genomic and proteomic approaches to understand the genetic predispositions and molecular pathways involved.

Effects of prophylactic propylene glycol administration at calving on subclinical ketosis in Holstein dairy cows

Forty-four holstein dairy cows were randomly enrolled in the treatment group (group 1, n=19) or control group (group 2, n=25) at calving. Group 1 received prophylactic propylene glycol treatment (PPGT) (300 ml/cow, beginning at calving, total 3 days). The group 2 remained untreated. All animals were tested on blood and milk beta-hydroxybutyric acid (BHBA) at postpartum week 2 and 4 (PW2 and 4), body condition scores (BCS), average daily milk production (ADMP) and postpartum health disorders were monitored in 90 days in milk (90 DIM). The incidence of subclinical ketosis (SCK) was 8% and 4% in blood test (BHBA≥1.2 mmol/L) and by 12% and 24% in milk test (BHBA≥200 µmol/L) in group 2 at PW2 and PW4 respectively. SCK was not detected in group 1. The study cows lost BCS on postpartum days 30 and 60. ADMP was significantly different (P

Plasma and milk metabolomics profiles in dairy cows with subclinical and clinical ketosis

Ketosis, a commonly observed energy metabolism disorder in dairy cows during the peripartal period, is distinguished by increased concentrations of β-hydroxybutyrate (BHB) in blood. This condition has a negative impact on milk production and quality, causing financial losses. An untargeted metabolomics approach was performed on plasma samples from cows between 5 and 7 DIM diagnosed as controls (CON, BHB <1.2 mM, n = 30), subclinically ketotic (SCK, 1.2 < BHB <3.0 mM, n = 30), or clinically ketotic (CK, BHB >3.0 mM, n = 30). Cows were selected from a commercial farm of 214 Holstein cows (average 305-d yield in the previous lactation of 35.42 ± 7.23 kg/d; parity, 2.41 ± 1.12; body condition score, 3.1 ± 0.45). All plasma and milk samples (n = 90) were subjected to Liquid Chromatography-Mass Spectrometry (LC-MS)-based metabolomic analysis. Statistical analyses was performed using the Graph Pad Prism 8.0, MetaboAnalyst 4.0 and R packages (version 4.1.3). Compared with the CON group, both SCK and CK groups had greater milk fat, freezing point, and fat-to-protein ratio and lower milk protein, lactose, solids-nonfat, and milk density. Within 21 d after calving, compared with CON, the SCK group experienced a reduction of 2.65 kg/d in milk yield, while the CK group experienced a decrease of 7.7 kg/d. Untargeted metabolomics analysis facilitated the annotation of a total of 5,259 and 8,423 metabolites in plasma and milk. Differentially affected metabolites were screened in CON vs. SCK, CON vs. CK, and SCK vs. CK (unpaired t-test, False discovery rate <0.05; and absolute value of log(2)-fold change >1.5). A total of 1,544 and 1,888 differentially affected metabolites were detected in plasma and milk. In plasma, glycerophospholipid metabolism, pyrimidine metabolism, tryptophan metabolism, sphingolipid metabolism, amino sugar and nucleotide sugar metabolism, phenylalanine metabolism, steroid hormone biosynthesis were identified as significant pathways. Weighted gene co-expression network analysis (WGCNA) indicated that tryptophan metabolism is a key pathway associated with the occurrence and development of ketosis. Increases in 5-Hydroxytryptophan and decreases in kynurenine and 3-indoleacetic acid in SCK and CK were suggestive of an impact at the gut level. The decrease of most glycerophospholipids indicated that ketosis is associated with disordered lipid metabolism. For milk, pyrimidine metabolism, purine metabolism, pantothenate and CoA biosynthesis, amino sugar and nucleotide sugar metabolism, nicotinate and nicotinamide metabolism, sphingolipid metabolism, fatty acid degradation were identified as significant pathways. The WGCNA indicated that purine and pyrimidine metabolism in plasma was highly correlated with milk yield during the peripartal period. Alterations in purine and pyrimidine metabolism characterized ketosis, with lower levels of these metabolites in both milk and blood underscoring reduced efficiency in nitrogen metabolism. Our results may help to establish a foundation for future research investigating mechanisms responsible for the occurrence and development of ketosis in peripartal cows.