Subclinical Ketosis Research Articles

Results of Screening of Goats in Diagnosing of Ketosis

Obtaining high‐quality and environmentally friendly livestock products must be provided with healthy livestock of animals. With ketosis, all types of metabolism are disturbed, the diagnosis and prevention of its various forms in ruminants are relevant. Dairy goats, like cattle, are prone to ketosis. Many representatives of dairy goat farms mistakenly believe that ketosis is not common among the dairy goat population. However, our initial studies of the prevalence of ketonemia among the population showed that at least 40% of animals in a herd had increased the concentration of ketone bodies in the blood. This fact indicates subclinical ketosis, which is promising for further research. In general, studies of metabolic disorders in dairy goats are not widespread; in this direction, the study of the metabolism of cattle occupies a leading position. The research was carried out on one of the goat farms in the North‐West region of Russia. We examined 5% of the total amount of animals at the farm (n=45). Goats were randomly selected, all from the milk group, from 1 to 8 lambing/lactation. Blood was obtained, and the concentration of β‐ketones was determined using a FreeStyle Optium Neo glucometer, researched the amount of total protein and its fractions, enzymes, lipids, and bilirubin RL‐Clima 15. Analysis of the study results and their comparison with the indicators of the referents revealed the following changes: the concentration of β‐ketones in 22 goats (48%) was in the range of 0.6 ‐ 0.9 mmol / L. Further, it was determined: the concentration of total protein was 13% higher than the limiting reference value and amounted to 86.4 ± 4.2 g / l, while no signs of dysproteinemia were observed. The urea level was also higher by 26.8% ‐ 9.29 ± 1.45 mmol / L. The activity of blood enzymes was within the normative values. The total bilirubin concentration was 10.5% higher than the reference value ‐ 5.7 ± 0.25 mmol / L, and the level of cholesterol was within the physiological range ‐ 2.11 ± 0.27 mmol / L. A violation of the calcium‐phosphorus ratio was noted. However, the concentrations of these substances were included in the reference values. Thus, it was determined that subclinical ketosis is prevalent up to about 50% among the goat population on a particular farm. Sick animals have protein metabolism disorders ‐ hyperproteinemia without dysproteinemia, and pigment metabolism disorders ‐ hyperbilirubinemia. It should be noted that no violations in the behavior and general condition of the animals were observed, which indicates precisely the subclinical form or stage of the disease. Timely prevention will reduce the percentage of sick animals.

Subclinical ketosis: Prevalence and some risk factors in cross breed and imported breed dairy cows in Mosul, Iraq

Although ongoing control strategies are attempting to minimize certain metabolic disorders such as hyperketonemia, factors such as genetics, management, environment, and geography still present a potential challenge to the achievement of the desired results. In light of this, this study was conducted (i) to determine the prevalence of subclinical and clinical ketosis based on the concentration of ketone bodies (BHB) using ELISA Kit, and (ii) to examine a number of epidemiological risk factors related to the conditions. From October, 2020 to the January, 2021. A 80 blood samples were collected in a cross-sectional survey in different areas of Mosul. Data included age, management systems, parity, and origin. At the cut-off point ≥1.2-≤ 1.4 mmol / L, the prevalence of subclinical ketosis was 27.5% and for clinical ketosis it was 5%. Results revealed that the animals aged > 3 years were more at risk for subclinical ketosis compared to animals aged ≤ 3 years RR = 2.6471. Multiparous animals were more at risk compared to the primiparous (two or less births) RR = 2.0652. No significant difference between the animals’ origins RR = 0.6863. Indoor and outdoor system animals were more at risk for subclinical ketosis compared to the indoor system only RR = 2.1389. In conclusion, hyperketonemia is prevalent in Mosul. Certain risk factors should be taking into account when planning control programs. Attention should be paid to diagnosis, management, and control of this disease during the risk stage to avert economic loss to the dairy farmers.

Estimating the burden of multiple endemic diseases and health conditions using Bayes’ Theorem: A conditional probability model applied to UK dairy cattle

The Global Burden of Animal Diseases (GBADs) is an international collaboration aiming, in part, to measure and improve societal outcomes from livestock. One GBADs objective is to estimate the economic impact of endemic diseases in livestock. However, if individual disease impact estimates are linearly aggregated without consideration for associations among diseases, there is the potential to double count impacts, overestimating the total burden. Accordingly, the authors propose a method to adjust an array of individual disease impact estimates so that they may be aggregated without overlap. Using Bayes’ Theorem, conditional probabilities were derived from inter-disease odds ratios in the literature. These conditional probabilities were used to calculate the excess probability of disease among animals with associated conditions, or the probability of disease overlap given the odds of coinfection, which were then used to adjust disease impact estimates so that they may be aggregated. The aggregate impacts, or the yield, fertility, and mortality gaps due to disease, were then attributed and valued, generating disease-specific losses. The approach was illustrated using an example dairy cattle system with input values and supporting parameters from the UK, with 13 diseases and health conditions endemic to UK dairy cattle: cystic ovary, disease caused by gastrointestinal nematodes, displaced abomasum, dystocia, fasciolosis, lameness, mastitis, metritis, milk fever, neosporosis, paratuberculosis, retained placenta, and subclinical ketosis. The diseases and conditions modelled resulted in total adjusted losses of £ 404/cow/year, equivalent to herd-level losses of £ 60,000/year. Unadjusted aggregation methods suggested losses 14–61% greater. Although lameness was identified as the costliest condition (28% of total losses), variations in the prevalence of fasciolosis, neosporosis, and paratuberculosis (only a combined 22% of total losses) were nearly as impactful individually as variations in the prevalence of lameness. The results suggest that from a disease control policy perspective, the costliness of a disease may not always be the best indicator of the investment its control warrants; the costliness rankings varied across approaches and total losses were found to be surprisingly sensitive to variations in the prevalence of relatively uncostly diseases. This approach allows for disease impact estimates to be aggregated without double counting. It can be applied to any livestock system in any region with any set of endemic diseases, and can be updated as new prevalence, impact, and disease association data become available. This approach also provides researchers and policymakers an alternative tool to rank prevention priorities.

METHODS OF DIAGNOSIS OF KETOSIS AND DISEASES OF REPRODUCTIVE FUNCTION IN HIGHLY PRODUCTIVE COWS

The article presents methods of express diagnostics and a laboratory method for determining the presence of ketone bodies in blood and urine. The research was carried out in 2018-2019 at the Department of "Veterinary Medicine" of NJSC «Kazakh Agrotechnical University named after S. Seifullin», in the farms of Akmola, Pavlodar and North Kazakhstan regions. The objects of research are cows of Holstein black-and-white and Simmental breeds (n=60) of the first and second lactation, during the early and late start, as well as in the first two weeks after calving. All breeding stock, taking into account the level of ketone bodies, was divided into groups: up to 1 mmol/l – clinically healthy, 1-2,5 mmol/l – subclinical ketosis, more than 2,5 mmol/l – clinically pronounced ketosis. Diagnosis of ketosis and diseases of reproductive function was carried out taking into account the early and late start, as well as the timing after calving cows. Ketone bodies in urine were determined in a qualitative and semi-quantitative way by the equipment of Keto-txt and Girul, Dirul Industrial Co., LTD and according to the Lestrade test method in laboratory conditions. The effectiveness of diagnostic methods for clinical and subclinical ketosis is 85-90%, the laboratory method of urine according to Lestrade is 87%, ultrasound examination of the reproductive system is 80%, rectal examination is 85-93%.

Metabolic profiling of serum and urine in lactating dairy cows affected by subclinical ketosis using proton nuclear magnetic resonance spectroscopy.

Ketosis is associated with high milk yield during lactating or insufficient feed intake in lactating dairy cows. However, few studies have been conducted on the metabolomics of ketosis in Korean lactating dairy cows. The present study aimed to investigate the serum and urine metabolites profiling of lactating dairy cows through proton nuclear magnetic resonance (1H-NMR) spectroscopy and comparing those between healthy (CON) and subclinical ketosis (SCK) groups. Six lactating dairy cows were categorized into CON and SCK groups. All experimental Holstein cows were fed total mixed ration. Serum and urine samples were collected from the jugular vein of the neck and by hand sweeping the perineum, respectively. The metabolites in the serum and urine were determined using 1H-NMR spectroscopy. Identification and quantification of metabolites was performed by Chenomx NMR Suite 8.4 software. Metabolites statistical analysis was performed by Metaboanalyst version 5.0 program. In the serum, the acetoacetate level was significantly (p < 0.05) higher in the SCK group than in the CON group, and whereas acetate, galactose and pyruvate levels tended to be higher. CON group had significantly (p < 0.05) higher levels of 5-aminolevulinate and betaine. Indole-3-acetate, theophylline, p-cresol, 3-hydroxymandelate, gentisate, N-acetylglucosamine, N-nitrosodimethylamine, xanthine and pyridoxine levels were significantly (p < 0.05) higher in the urine of the SCK group than that in the CON group, which had higher levels of homogentisate, ribose, gluconate, ethylene glycol, maltose, 3-methyl-2-oxovalerate and glycocholate. Some significantly (p < 0.05) different metabolites in the serum and urine were associated with ketosis diseases, inflammation, energy balance and body weight. This study will be contributed useful a future ketosis metabolomics studies in Korea.

Differences in the serum metabolome profile of dairy cows according to the BHB concentration revealed by proton nuclear magnetic resonance spectroscopy (1H-NMR)

The mobilization of body reserves during the transition from pregnancy to lactation might predispose dairy cows to develop metabolic disorders such as subclinical ketosis or hyperketonemia. These conditions are not easily identifiable and are frequently related to other diseases that cause economic loss. The aim of this study was to evaluate the serum metabolome differences according to the β-hydroxybutyrate (BHB) concentration. Forty-nine Holstein Friesian dairy cows were enrolled between 15 and 30 days in milk. According to their serum BHB concentration, the animals were divided into three groups: Group 0 (G0; 12 healthy animals; BHB ≤ 0.50 mmol/L); Group 1 (G1; 19 healthy animals; 0.51 ≤ BHB < 1.0 mmol/L); and Group 2 (G2; 18 hyperketonemic animals; BHB ≥ 1.0 mmol/L). Animal data and biochemical parameters were examined with one-way ANOVA, and metabolite significant differences were examined by t-tests. Fifty-seven metabolites were identified in the serum samples. Thirteen metabolites showed significant effects and seemed to be related to the mobilization of body reserves, lipids, amino acid and carbohydrate metabolism, and ruminal fermentation.

Activation of Transcription Factor EB Is Associated With Adipose Tissue Lipolysis in Dairy Cows With Subclinical Ketosis.

Excessive lipid mobilization for adipose tissue caused by severe negative energy balance is the pathological basis for subclinical ketosis (SCK) in dairy cows. In non-ruminants, transcription factor EB (TFEB) was reported to play a role in the regulation of lipid catabolism, but its role in the control of lipolysis in the bovine is unknown. The present study aimed to determine whether the enhanced TFEB transcriptional activity contributes to lipolysis of adipose tissue in SCK cows, and to explore the possibility of establishing a therapeutic strategy by using TFEB as a target to control lipolysis. Thirty cows with similar lactation number (median = 3, range = 2–4) and days in milk (median = 6 d, range = 3–9) were selected into a healthy control (n = 15) and SCK (n = 15) group, and used for subcutaneous adipose tissue biopsies and blood sampling. Adipocytes from healthy Holstein calves were used as a model for in vitro studies involving treatment with 10 μM isoproterenol (ISO) for 0, 1, 2 and 3 h, 250 nM of the TFEB activator Torin1 for 3 h, or used for transfection with TFEB small interfering RNA for 48 h followed by treatment with 10 μM ISO for 3 h. Compared with healthy cows, adipose tissue in SCK cows showed increased lipolysis accompanied by enhanced TFEB transcriptional activity. In vitro, ISO and Torin1 treatment increased lipolysis and enhanced TFEB transcriptional activity in calf adipocytes. However, knockdown of TFEB attenuated ISO-induced lipolysis in adipocytes. Overall, these findings indicated that enhanced transcriptional activity of TFEB may contribute to lipolysis of adipose tissue in dairy cows with SCK. The regulation of TFEB activity may be an effective therapeutic strategy for controlling overt lipolysis in ketotic cows.

Differentiation of Subclinical Ketosis and Liver Function Test Indices in Adipose Tissues Associated With Hyperketonemia in Postpartum Dairy Cattle.

Past studies suggested that during early lactation and the transition period, higher plasma growth hormone (GH) levels in subclinical ketosis (SCK) might involve the initiation of body adipose tissues mobilization, resulting in metabolic disorders in ruminants particularly hyperketonemia. The upregulated GH mRNA expression in adipose tissue may take part in the adipolysis process in SCK-affected cows that paves a way for study further. This study aimed to characterize the plasma levels of GH, β-hydroxybutyrate acid (BHBA) and non-esterified fatty acid (NEFA) and glucose (GLu) in ketotic cows and healthy control (CON) cows; to measure the liver function test (LFT) indices in ketotic and healthy CON cows, and finally the quantitative real-time PCR (qRT-PCR) assay of candidate genes expressed in adipose tissues of ketotic and healthy CON cows during 0 to 7 week postpartum. Three experiments were conducted. Experiment-1 involved 21 Holstein cows weighing 500–600 kg with 2–5 parities. Results showed that GH, BHBA, and NEFA levels in ketotic cows were significantly higher and the GLu level significantly lower. Pearson's correlation analysis revealed a significant positive correlation of GH with BHBA, NEFA, and GLu in ketotic and healthy CON cows. In experiment-2, dynamic monitoring of LFT indices namely, alanine aminotransferase (ALT), aspartate aminotransferase (AST), γ-glutamyl transpeptidase (GGT), total bilirubin (TBIL), direct bilirubin (DBIL), total protein (TP), albumin (ALB), globulin (GLOB) and albumin/globulin (A/G) were examined. The TBIL, DBIL, and GGT indices were significantly higher in ketotic cows and TP was significantly lower. In experiment-3, mRNA expression levels of GHR and peroxisome-proliferator-activated receptor alpha (PPARα) genes in adipose tissue were significantly upregulated in ketotic cows. However, the mRNA expression of insulin-like growth factor-I (IGF-1), insulin-like growth factor-I receptor (IGF-1R), and sterol regulatory element-binding protein-1c (SREBP-1c) genes in adipose tissue were downregulated in ketotic cows. Our study concluded that during postpartum, higher plasma GH levels in SCK cows might involve the initiation of body adipose tissue mobilization, resulting in hyperketonemia.

The Use of Multilayer Perceptron Artificial Neural Networks to Detect Dairy Cows at Risk of Ketosis.

Simple SummaryKetosis is a serious metabolic disease in high-yield dairy cows, that affects productive herds throughout the world. Subclinical ketosis is one of the most dominant metabolic disorders in dairy herds during early lactation, so early detection and prevention are important for both economic and animal welfare reasons. Neural networks, which offer a high degree of accuracy in predicting various phenomena and processes where there is no clear causal correlation or there are no rules that allow the establishment of a logical cause-and-effect relationship, can be used to address problems related to prediction, classification, or control. A Multi-Layer perceptron (MLP) is a feedforward artificial neural network model that takes input data for a set of proper output. This study investigated the performance of four algorithms used to train MLP networks. The experimental results demonstrate that the MLP network model improved the accuracy of process recognition of subclinical ketosis in dairy cows. The received artificial model’s results were saved in the predictive model markup language (PMML) and can be used to describe the learning set, the algorithm used in the data mining application and related information.Subclinical ketosis is one of the most dominant metabolic disorders in dairy herds during lactation. Cows suffering from ketosis experience elevated ketone body levels in blood and milk, including β-hydroxybutyric acid (BHB), acetone (ACE) and acetoacetic acid. Ketosis causes serious financial losses to dairy cattle breeders and milk producers due to the costs of diagnosis and management as well as animal welfare reasons. Recent years have seen a growing interest in the use of artificial neural networks (ANNs) in various fields of science. ANNs offer a modeling method that enables the mapping of highly complex functional relationships. The purpose of this study was to determine the relationship between milk composition and blood BHB levels associated with subclinical ketosis in dairy cows, using feedforward multilayer perceptron (MLP) artificial neural networks. The results were verified based on the estimated sensitivity and specificity of selected network models, an optimum cut-off point was identified for the receiver operating characteristic (ROC) curve and the area under the ROC curve (AUC). The study demonstrated that BHB, ACE and lactose (LAC) levels, as well as the fat-to-protein ratio in milk, were important input variables in the network training process. For the identification of cows at risk of subclinical ketosis, variables such as BHB and ACE levels in milk were of particular relevance, with a sensitivity and specificity of 0.84 and 0.61, respectively. It was found that the back propagation algorithm offers opportunities to integrate artificial intelligence and dairy cattle welfare within a computerized decision support tool.

Metabolite Comparison between Serum and Follicular Fluid of Dairy Cows with Inactive Ovaries Postpartum.

Simple SummaryAlthough the milk production of dairy cows has increased rapidly in recent decades, the reproductive performance of dairy cows has gradually declined. In modern intensive dairy farms, prevention and treatment of inactive ovaries has become an important challenge of reproduction disorders during early lactation. Our aim is to screen out metabolites and metabolic pathways related to inactive ovaries through serum and follicular fluid metabolomics. We found that the changes in serum and follicular fluid were mainly enriched in nine metabolic pathways. In serum, these included d-glutamine and d-glutamate metabolism, alanine, aspartic and glutamate metabolism, arginine and proline metabolism, pentose and glucuronate interconversions, and glycerophospholipid metabolism. In follicular fluid, they were valine, leucine, and isoleucine biosynthesis; arachidonic acid metabolism; glycerophospholipid metabolism; starch and sucrose metabolism; phenylalanine metabolism; and pentose and glucuronate interconversion. The common metabolic pathways of disease-related serum and follicular fluid were pentose and glucuronate interconversions and glycerophospholipid metabolism. This research will provide a theoretical basis for exploring the causes of inactive ovaries and provide new ideas for the prevention and treatment of inactive ovaries in the future.Inactive ovaries (IO) accounts for 50% of ovarian disease in postpartum dairy cows, which seriously affects their reproductive efficiency. To investigate the metabolic changes in the serum and follicular fluid of dairy cows with IO during lactation, six estrus (E) cows and six IO cows at 50 to 55 days in milk were selected based on B ultrasonic detection and clinical manifestations. The differential metabolites in serum and follicular fluid between the E cows and IO cows were identified by ultra-high-pressure liquid chromatography–quadrupole time-of-flight mass spectrometry, combined with multidimensional statistical methods. The results showed that dairy cows with IO were in a subclinical ketosis status where beta-hydroxybutyrate (BHB) exceeded 1.20 mmol/L, 14 differential metabolites in the serum of IO cows included 10 increased metabolites and 4 decreased metabolites, and 14 differential metabolites in the follicular fluid of IO cows included 8 increased metabolites and 6 decreased metabolites. These differential metabolites mainly involved nine metabolic pathways. The common enrichment pathway of different metabolites in serum and follicular fluid were glycerophospholipid metabolism and pentose and glucuronate interconversions. In conclusion, there were significant differences in the differential metabolites and enrichment pathways between serum and follicular fluid of IO cows, implying that there were complex changes in blood metabolism and local follicular metabolism of IO cows, whose interactions need further investigation.

Relationship between Postpartum Metabolic Status and Subclinical Endometritis in Dairy Cattle.

Simple SummaryProper reproductive efficiency is decisive to achieving adequate profitability in dairy farms. However, uterine pathologies such as subclinical endometritis (SE) play a primary role in the decline of reproductive performance. This disease impairs reproductive function, and its incidence may reach up to 34% during the first seven weeks after calving. Consequently, identifying the predisposing factors and diagnosing this pathology as early as possible is mandatory to minimize the impact on the profitability of the farms. Several metabolic alterations postpartum have been related to the occurrence of SE, so our objective was to identify which alterations act as risk factors for SE. Uterine and blood samples and data from 94 Holstein cows were collected 30–45 days after calving. Our results showed that serum levels of β-hydroxybutyrate acid (BHBA), albumin, and urea are related to the incidence of SE, being BHBA a predisposing factor and albumin and urea protective factors. Therefore, these metabolites should be carefully considered during the postpartum period as indicators of SE. Additionally, preventive measures aimed to control these alterations may be useful to prevent SE.The aim of this study was to verify the importance of postpartum serum levels of certain metabolic markers as risk factors for subclinical endometritis (SE). Ninety-four Holstein cows were included in the study, and examinations were carried out between 30–45 days postpartum. Rectal palpation, vaginoscopy, transrectal ultrasound, endometrial cytology, and blood sample collections were performed. The percentage of polymorphonuclear neutrophils (%PMN) on the endometrium was evaluated, as well as serum levels of glucose, cholesterol, triglyceride, albumin, hepatic enzymes, urea, non-esterified fatty acids (NEFA), and β-hydroxybutyrate acid (BHBA). Samples with ≥8% PMN were classified as positive to subclinical endometritis. According to the serum levels of BHBA, cows were classified as clinical ketosis (>2.6 mmol/L), subclinical ketosis (1.2–2.6 mmol/L), and healthy (<1.2 mmol/L). Additionally, body condition score, parity, date of last labor, peripartum issues, insemination date, date of pregnancy diagnosis and milk production information were collected. Data were analyzed using a multiple regression analysis. The results showed that as serum levels of BHBA rose, also did the %PMN, so that up to 60% of cows with clinical ketosis suffered from SE. On the other hand, the %PMN fell as serum levels of urea and albumin increased. Consequently, good postpartum management practices and early detection of metabolic alterations are necessary measures to control predisposing factors and reduce the incidence of SE.

Prevalence, risk factors and metabolic predictors associated with subclinical ketosis on dairy cattle in Batna, Eastern of Algeria

Economic impact of subclinical ketosis (SCK) in dairy herds resulted from a decreased productivity, reproductive performances, and health condition. In Algeria, data availability regarding the prevalence of SCK in dairy cows were limited. The present study is a contribution in that direction and aimed to determine the prevalence of SCK between 2 and 50 days postpartum whereas investigating herd- and cow-level risk factors and the metabolic profile associated, on two bovine populations imported and native. The study was conducted with 16 typical representative local dairy cow herds (n = 100; 2–12 years old; located at Merouana, Batna Province) with data collected in a period comprised between August and February of the following year. Concentration of cows’ plasma beta-hydroxybutyrate (BHBA) was used for the diagnosis, with values equal or higher than 1.2 mmol/L chosen as the threshold for SCK detection. Other key associated zootechnical parameters such as body condition score (BCS) were monitored at the same sampling points. Logistic regression analyses were carried out to characterize association between parameters. The average SCK prevalence was 9% and resulted highest during first- and second-week postcalving, with a peak in the first week. The herd size (< 30 lactating cows) and BCS (≤ 3) were risk factors associated with SCK prevalence. At the cow-level, risks of SCK increased in thin individuals (P = 0.058; odds ratio (OR) = 5.00, 95% confidence interval (CI) = 0.95–26.32). At the herd-level, risks of SCK increased in smaller frame size herds (P = 0.047; OR = 8.76, 95% CI = 1.03–74.49). Most cases of SCK were observed in native cows compared to imported. Interestingly, linear stepwise regression showed a good relationship between BHBA and total bilirubin (P = 0.0001; R2 = 0.59). There were significant differences between healthy and ketotic cows regarding plasma concentration of bilirubin (P = 0.003), glucose (P = 0.001), alanine aminotransferase (ALT) (P = 0.008), creatinine (P = 0.007), and sodium (P = 0.02). According to receiver operating characteristic (ROC) curve analysis, the best parameter for SCK prediction is total bilirubin with more than 4.09 g/L as a cutoff point (77.78% sensitivity and 94.51% specificity). According to metabolic profile and feeding data we suggest that most cases of SCK observed in our study are type I. Further studies are required to validate other metabolic predictors for SCK and investigate adaptation of imported cows in semiarid regions in Algeria.

Predicting ketosis during the transition period in Holstein Friesian cows using hematological and serum biochemical parameters on the calving date

Ketosis often occurs during the postpartum transition period in dairy cows, leading to economic and welfare problems. Previously, ketosis was reported to be associated with hematological and serum biochemical parameters. However, the association between the parameters on the calving date and ketosis during the postpartum transition period remains unclear. This study aimed to investigate this association. Blood samples were collected from the jugular vein of Holstein cows on the calving date and β-hydroxybutyrate was tested once every 3 days (8 times in 21 days). The cows were divided into three groups: non-ketosis, subclinical ketosis, and clinical ketosis. The clinical ketosis group significantly had the highest values of mean corpuscular volume, mean corpuscular hemoglobin, β-hydroxybutyrate, non-esterified fatty acids, and total bilirubin, but the lowest values of red cell distribution width, the counts of white blood cell, monocyte, and eosinophil, albumin, alanine transaminase, lactate dehydrogenase, and amylase. In contrast, the non-ketosis group showed the opposite results (p < 0.05). In conclusion, these parameters are associated with the development and severity of ketosis. Our findings suggest that these parameters on the calving date may be useful indicators to identify dairy Holstein cow susceptible to ketosis during the transition period.

Global prevalence of subclinical ketosis in dairy cows: A systematic review and meta-analysis

The objectives of this study were to estimate the global prevalence of SCK in dairy cows, to identify optimum thresholds for BHB determining, to explore heterogeneity for some particular characteristics (continents, diagnosis techniques, cut-off, sample type, parity, breed, and housing), and to determine the SCK prevalence trend. SCK prevalence levels in dairy cows from various countries were calculated using bibliographic databases up until May 17, 2021 to screen titles and abstracts for possible eligibility. Thirty-eight studies from six continents were then included using a random-effects model to calculate the pooled SCK prevalence. Subgroup meta-analyses were performed to assess the heterogeneity of the prevalence for the above characteristics. Meta-regression and cumulative meta-analysis were used to assess the SCK prevalence trends. The results showed that the global prevalence of SCK in dairy cows was 22.7% [95% CI 21.2–24.3%]. No significant differences in SCK prevalence were observed among continents, diagnostic techniques, cut-off values—≥1.0, ≥1.2 and ≥1.4 mmol/L, sample types—milk and blood, and parities. However, the prevalence in Holsteins (19.8%) was significantly lower than other mixed breeds (23.7%). The SCK prevalence was significantly higher in indoor barns (27.8%) than in pasture and unspecified housing. In meta-regression, the SCK prevalence was not associated with study years or days in milk. The cumulative evidence suggested that the prevalence of SCK in dairy cows was associated with a wide range of risk factors. This study illustrates that the global prevalence of SCK in dairy cows is quite high; therefore, there is an urgent need to globally reduce the SCK prevalence in dairy farms.

Free fatty acids promote degranulation of azurophil granules in neutrophils by inducing production of NADPH oxidase–derived reactive oxygen species in cows with subclinical ketosis

Subclinical ketosis (SCK) in dairy cows, a common metabolic disorder during the peripartal period, is accompanied by systemic inflammation. Excessive release of azurophil granule (AG) contents during degranulation of polymorphonuclear neutrophils (PMN) could contribute to systemic inflammation in SCK cows. Although the increase in blood free fatty acids (FFA) in SCK cows may promote AG degranulation from PMN, the underlying mechanisms are unclear. Thirty multiparous cows (within 3 wk postpartum) with similar lactation numbers (median = 3, range = 2-4) and days in milk (median = 6, range = 3-15) were classified based on serum β-hydroxybutyrate (BHB) level as control (n = 15, BHB < 0.6 mM) or SCK (n = 15, 1.2 mM < BHB < 3.0 mM). Cows with SCK had greater levels of serum haptoglobin, serum amyloid A, IL-1β, IL-6, IL-8 and tumor necrosis factor-α. These proinflammatory factors had strong positive correlations with myeloperoxidase (MPO), a marker protein of PMN AG, whose content was greater in the serum of SCK cows. Both the number of AG and the protein abundance of MPO were lower in PMN isolated from SCK cows. Additionally, we found a greater ratio of blood CH138A+/CD63high cells and greater mean fluorescence intensity of CD63 on the PMN membrane, further confirming the greater degree of AG degranulation in cows with SCK. In vitro FFA dose response (0, 0.3, 0.6, 1.2, and 2.4 mM for 4 h) and time course (0, 0.5, 1, 2, and 4 h with 0.6 mM) experiments were performed on PMN isolated from control cows. The increase in MPO content in extracellular supernatant resulting from those experiments led to the selection of 0.6 mM FFA for 1 h duration as conditions for subsequent studies. After FFA treatment, release of intracellular MPO was increased along with increased levels of CD63 mean fluorescence intensity on the PMN membrane, confirming that FFA promoted degranulation of AG. In addition, FFA treatment increased reactive oxygen species (ROS) production by PMN, an effect that was attenuated by incubation with diphenyleneiodonium chloride (DPI), a NADPH oxidase-derived ROS inhibitor. The mitochondrial-derived ROS inhibitor carbonyl cyanide 4-(trifluoromethoxy) phenylhydrazone (FCCP) did not affect ROS in response to FFA treatment. Treatment with FFA increased p47 phosphorylation and mRNA abundance of NCF1, NCF2, and CYBB in PMN. Furthermore, DPI, but not FCCP, dampened the degranulation of PMN AG induced by FFA in vitro. These data suggested that the degranulation of AG in PMN induced by FFA was mediated by NADPH oxidase-derived ROS. As verified ex vivo, PMN from SCK cows had greater levels of ROS, phosphorylation of p47, and mRNA abundance of NCF1, NCF2, and CYBB. Overall, the present study revealed that high blood concentrations of FFA in SCK cows induce the production of NADPH oxidase-derived ROS, thereby promoting degranulation of AG in PMN. The stimulatory effect of FFA on the release of AG content during degranulation, especially MPO, provides a new insight into the systemic inflammation experienced by peripartal cows with SCK.

Subclinical ketosis: Prevalence and risk factor analysis in early lactation dairy cattle of Thrissur district

Present study was conducted in Kerala Veterinary and Animal Sciences University farms, various organized and unorganised farms and individual households in Thrissur district during the period of May 2021 to April 2022 with the objectives of estimation of prevalence of subclinical ketosis (SCK) by screening dairy cows on 14th and 28th day post-partum. Multiparous cows with a blood beta-hydroxy butyrate (BHB) value ≥ 1.2 mmol/L without any overt clinical signs were considered as positive for SCK. One hundred and fourteen animals from second to seventh lactation were screened for SCK. Prevalence of SCK was 23.7 per cent on 14th day and 14.04 per cent on 28th day with a total prevalence of 27.2 per cent. Occurrence of SCK was highest in animals in second lactation (54.8 per cent) with more occurrence in crossbred Holstein Friesian cows. Maximum number of positive cases were observed in dairy cows of age between four to six years (54.8 per cent). Body condition score of diseased animals were higher than normal animals. Prevalence was highest in animals calved between months of July to September and 58.1 per cent of affected animals had male calf. Concurrent abnormalities during transition period were also assessed based on clinical examination and farm records.

Incidence of Subclinical Ketosis and Evaluation of Associated Risk Factors in Early Lactating Dairy Buffaloes

Incidence of Subclinical Ketosis and Evaluation of Associated Risk Factors in Early Lactating Dairy Buffaloes

Blood and Milk Beta-hydroxybutyric Acid Concentrations in Diff erent Dairy Cattle Breeds and Association of Subclinical Ketosis with Postpartum Health Disorders, Culling Rate, Body Condition Score, Parity and Milk Production in Holstein

Blood and Milk Beta-hydroxybutyric Acid Concentrations in Diff erent Dairy Cattle Breeds and Association of Subclinical Ketosis with Postpartum Health Disorders, Culling Rate, Body Condition Score, Parity and Milk Production in Holstein

Subclinical ketosis as a factor reduction of reproductive indicators of high productive cows

Purpose: The aim of the research is to study the effect of subclinical ketosis (SCK) on the effectiveness of artificial insemination of highly productive dairy cows in the postpartum period.Materials and methods. The studied animals were selected according to the principle of conditional analogs and were divided into 2 groups of 6 animals each. Group 1 - animals with a blood concentration of 3- hydroxybutyrate˂ 1.0 mmol / l, group 2 - animals with subclinical ketosis with a concentration of 3-hydroxybutyrate in the blood in the range of 1.0-1.4 mmol / l. Blood was collected from animals on the 5th, 15th and 33-35th days after calving. In the obtained blood serum samples, the content of total protein, albumin, urea, glucose, cholesterol, triglycerides, calcium, phosphorus, magnesium, creatinine, total bilirubin, the activity of transaminase enzymes and alkaline phosphatase was determined. The concentration of steroid hormones has been determined. Before artificial insemination, the animals were synchronized according to the pressinovsinh scheme.Results. Most of the biochemical blood parameters did not go beyond the reference values. By the time of hormonal stimulation of sexual heat in cows with SCK, a decrease in 3-hydroxybutyrate (˂ 1,0 mmol / l) and an increase in glucose (&gt; 3.0 mmol / l) were recorded. On the 15th day after calving, in the group with SCK, there was an increase in total bilirubin, cholesterol, triglycerides, and magnesium (p˂ 0,01). On the 15th day after calving, testosterone levels were 13% higher in the SCK group. By the time of synchronization, the level of progesterone in the group of animals with SCK was lower in comparison with the control group. In the group of animals with SCK, the lowest reproductive indices were recorded: the insemination index in the group with SSC was higher by 46%, the service period was 49 days longer, the interbody period was significantly longer by 68 days (p˂ 0,05).Conclusion. Subclinical ketosis, recorded on the 15th day after calving, helps to reduce the reproductive performance of high -yielding dairy cows. Normalization of the concentration of 3-hydroxybutyrate from subclinical to physiologically normal values, and subsequent hormonal synchronization do not reduce this negative effect of SCK.

EFECTOS DEL ESTADO ENERGÉTICO Y PROTEICO SOBRE PARÁMETROS REPRODUCTIVOS Y PRESENCIA DE CETOSIS SUBCLÍNICA EN VACAS BROWN SWISS POSPARTO

This study aimed to determine protein and energy metabolites in postpartum Brown Swiss cows and their effect on the presence of subclinical ketosis and reproductive parameters. One hundred cows from four farms in the Mantaro Valley (Junín, Perú) were grouped according to parity (1, 2, 3 and 4). The variables of the metabolic profile (total proteins, blood urea nitrogen [BUN], glucose, βhydroxybutyrate [B-HB]), milk production and body condition were estimated between 7 to 60 days postpartum in 7-day intervals. Likewise, the calving-first heat interval and the pregnancy rate at first service were recorded. The determination of subclinical ketosis was based on a semi-quantitative analysis by levels of B-HB in milk (µmol/l) using a commercial kit. BUN levels ranged between 11.74 and 15.92 mg/dl, being higher in fourth parity cows (p&lt;0.05). The averages of total protein (6.54-7.90 g/dl) were homogeneous between calvings. The glucose values presented an inverse response, being lower in cows of third and fourth parity compared to cows of first and second parity (p&lt;0.05). Similarly, the highest levels of milk production were observed in third parity cows (14.41 ± 5.42 l/d) and fourth parity (15.43 ± 4.36 l/d) with respect to first and second calvers (p&lt;0.05). Body condition was lower in cows with subclinical ketosis. The calving - first heat interval and the pregnancy rate at first service were lower in cows with subclinical ketosis (p&lt;0.05).