Β-hydroxybutyric Acid Concentrations Research Articles

Conventional and Doppler ultrasonographic diagnosis of subclinical pregnancy toxemia with altered serum cortisol and biochemical parameters in pregnant does

The pathogenesis and diagnosis of subclinical pregnancy toxemia (SCPT) remain elusive and need further investigation in pregnant does. Therefore, the aim of our study was to describe the typical properties of hepatic venous hemodynamics by Doppler ultrasonography. A total of 70 pregnant does were classified based on the blood serum concentrations of β-hydroxybutyric acid (βHBA), pregnant does were categorized into control group (βHBA concentrations <0.8 mmol/L; n = 40) and SCPT group (βHBA concentrations >0.8 mmol/L; n = 30). DRAMISKI 4vet slim diagnostic ultrasound scanner with B, M, and Doppler (color, power, pulsing wave) modes was used for diagnosis of SCPT. Total serum cortisol level was quantitative using chemiluminescent immunoassay. Serum glucose, triglycerides, cholesterol, HDL and LDL- cholesterol and LDH- cholesterol were measured by colorimetric and kinetic methods. Liver ultrasonography of does with SCPT had been shown mild fatty infiltration with rounded margin, which was characterized by hyperechoic area. There was a significant decrease in the values of portal vein diameter (PVD), portal vein area (PVA), portal mean velocity (PMV) and portal blood flow (PBF) in SCPT does compared to control pregnant does. PVD, PVA and PBF were negatively correlated with βHBA concentrations in does with SCPT (P < 0.05). PVD was inversely associated with serum cholesterol and triglycerides concentrations (P < 0.05). In conclusions, Doppler ultrasonography examinations of pregnant does with SCPT indicate abnormal hepatic variation. Reduced PVD, PVA, PMV and PBF together with increased βHBA concentrations could predict SCPT in does with fair sensitivity and specificity.

Effect of β-hydroxybutyrate acid on gene expression levels of antioxidant biomarkers and growth hormone-related genes in liver cell culture.

In dairy cattle, oxidative stress is a predominant problem associated with diseases and reproductive health issues. This study aimed to detect the variation in the antioxidant biomarkers by adding different concentrations of β-hydroxybutyric acid (BHBA) and sought to elucidate its effects on the gene expression levels of growth hormone (GH) and antioxidant biomarkers in bovine hepatocytes. Four antioxidant biomarkers, namely malondialdehyde (MDA), superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GSH Px) were evaluated using commercially available bovine ELISA kits. The expression levels of the bovine GH, its receptor (GHR), insulin-like growth factor (IGF), IGF-1, IGF-1 receptor, CAT, SOD, GSH-Px and β-actin (as a reference) genes in liver cell culture were determined by reverse transcriptase-PCR assay. With the increase of BHBA concentration and culture time, the activities of SOD, CAT, and GSH Px biomarkers in hepatocytes decreased. However, the content of MDA in hepatocytes increased gradually with the increase of hepatocyte culture time and BHBA concentration. The qPCR results revealed that after adding BHBA, gene expression levels of GSH-Px, SOD and IGF biomarkers in hepatocytes began to differ in the culture groups at 12 h, whereas the gene expression level of the CAT and GHR biomarkers in hepatocytes began to differ at 6 h. Quantitative PCR results showed that the BHBA significantly downregulated the expression levels of the GHR gene and CAT, GSH Px and SOD antioxidant biomarker genes.

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.

The Effect of Dietary Supplementation with Probiotic and Postbiotic Yeast Products on Ewes Milk Performance and Immune Oxidative Status.

The administration of yeast products as feed additives has been proven to beneficially affect animal productivity through energy, oxidative, and immune status improvement. This study evaluated a combination of Saccharomyces cerevisiae live yeast (LY) with yeast postbiotics (rich in mannan-oligosaccharides (MOS) and beta-glucans) and selenium (Se)-enriched yeast on ewes' milk performance and milk quality, energy and oxidative status, and gene expression related to their immune system during the peripartum period. Ewes were fed a basal diet (BD; F:C = 58:42 prepartum and 41:59 postpartum) including inorganic Se (CON; n = 27), the BD supplemented with a LY product, and inorganic Se (AC; n = 29), as well as the combination of the LY, a product of yeast fraction rich in MOS and beta-glucans, and organic-Se-enriched yeast (ACMAN; n = 26) from 6 weeks prepartum to 6 weeks postpartum. The β-hydroxybutyric acid concentration in the blood of AC and ACMAN ewes was lower (compared to the CON) in both pre- and postpartum periods (p < 0.010). Postpartum, milk yield was increased in the AC and ACMAN Lacaune ewes (p = 0.001). In addition, the activity of superoxide dismutase (p = 0.037) and total antioxidant capacity (p = 0.034) measured via the 2,2-Azino-bis (3-ethylbenzthiazoline-6-sulfonic acid) (ABTS) method was increased in the blood plasma of the ACMAN postpartum. Higher ABTS values were also found (p = 0.021), while protein carbonyls were reduced (p = 0.023) in the milk of the treated groups. The relative transcript levels of CCL5 and IL6 were downregulated in the monocytes (p = 0.007 and p = 0.026 respectively), and those of NFKB were downregulated in the neutrophils of the ACMAN-fed ewes postpartum (p = 0.020). The dietary supplementation of ewes with yeast postbiotics rich in MOS and beta-glucans, and organic Se, improved energy status, milk yield and some milk constituents, and oxidative status, with simultaneous suppression of mRNA levels of proinflammatory genes during the peripartum period.

Lactation performance, milk fat output, and nutrient digestibility responses to the addition of liquid molasses or yeast culture in dairy cows fed super-conditioned corn

Increased diet fermentability may decrease ruminal pH and fiber digestibility, and increase the flow of trans fatty acids (FA) to the lower tract ultimately leading to milk fat depression. We recently showed that feeding super-conditioned corn, a new method of corn processing (95°C for 6 min in super-conditioner) for ruminants has potential to the reduction in milk fat yield caused by changes in ruminal pH and increased trans FA in milk fat. Supplementing yeast culture (YC) and replacing starch with sugar sources in diet can counteract the negative effects of high fermentable diets by improving ruminal pH and milk fat output. This study aimed to evaluate the effect of feeding beet liquid molasses (LM) and YC on intake and total-tract digestibility of nutrients, milk yield and composition, ruminal fermentation, milk FA profile, and plasma concentrations of glucose, nonesterified FA, β-hydroxybutyric acid, and urea N in early-lactation dairy cows fed high-starch diets containing super-conditioned corn. Twelve primiparous and 18 multiparous Holstein cows (mean ± SD; 67 ± 12 d in milk and 42 ± 2.1 kg of milk at the beginning of the experiment) were blocked by parity, pre-experimental milk yield, and DIM. Cows were used in a randomized complete block design experiment with 14 d as covariate period and 37 d for the experimental period. The following dietary treatments were fed as total mixed rations: (1) control diet (CTRL = no YC or LM supplementation), (2) LM supplementation at 5% of the diet dry matter (MOL diet), and (3) CTRL supplemented with 10 g/d of YC (YST diet). Diets were formulated to be isonitrogenous and isoenergetic. Intake of nutrients and apparent total-tract digestibility of crude protein and starch did not change across treatments. In contrast, cows fed the YST diet had the greatest apparent total-tract digestibility of dry matter, organic matter and neutral detergent fiber. Compared with the CTRL diet, yield of 4% FCM increased by 2.4 and 1.8 kg in cows fed MOL or YST, respectively. The ruminal molar proportions of acetate and butyrate increased in cows fed the YST or MOL diets, respectively, but the proportion of ruminal propionate was not affected by treatments. Milk fat concentration increased by supplementing both LM and YC and the milk yield of total trans-18:1 dropped by 45% and 18% relative to CTRL with MOL or YST diets, respectively. While the MOL diet increased the milk proportion and yield of de novo FA, no treatment effects were observed for the proportion and yield of preformed FA in the milk fat. Apart from β-hydroxybutyric acid concentration in plasma, which was greatest in cows fed MOL, remaining blood metabolites were not affected by treatments. Overall, MOL and YST diets increased 4% FCM and milk fat concentration and reduced the proportion of total trans-18:1 FA in milk fat in cows fed a concentrate based on super-conditioned corn. These responses were associated with increased ruminal pH and the molar proportions of acetate and butyrate with feeding the MOL and YST diets.

Interaction between the Concentration of β-Hydroxybutyric Acid and the Content of Long-Chain Fatty Acids in the Early Stage of Lactation––Comparing Multiparous and Primiparous Cows

The aim of the experiment was to study the relationship between the age of cows, blood BHBA content, and CLA isomer (C18:2 cis9,trans11, CLA9; C18:2 trans10,cis12, CLA10) content during the first three weeks post-partum. For the experiment, 105 cows were selected from the entire herd and assigned to one of four groups: healthy primiparous (PH), healthy multiparous (MH) or ketotic primiparous (PK), ketotic multiparous (MK) based on their symptoms, and blood serum BHBA concentrations at 5 ± 2 days post-partum. Milk and blood samples were taken from the animals for a period of three weeks at weekly intervals on the same day. High levels of ketone bodies inhibit the activity of acetyl-CoA, thus decreasing the transport of acetyl-CoA, which may result in a decrease in CLA9 and CLA10 synthesis. Studies have shown that the age of the cows was an additional factor in determining the formation of CLA isomer levels during the early stage of lactation. The CLA9 content in the milk of PH cows in the first week of lactation was 32.75% higher than that of PK milk, while in MH milk, it was 67.7% higher than that of MK milk. The CLA10 content in the milk PH, when compared to the healthy groups, was 319% lower for primiparous cows. In summary, different reference limits in CLA9 and CLA10 content should be considered in the diagnosis of ketosis, taking into account, among other things, parity.

Effects of liquefied sake lees on growth performance and faecal and blood characteristics in Japanese Black calves.

Liquefied sake lees, a by-product of Japanese sake, is rich in Saccharomyces cerevisiae, proteins, and prebiotics derived from rice and yeast. Previous studies have reported that Saccharomyces cerevisiae fermentation products improved the health, growth, and faecal characteristics of preweaning calves. This study investigated the effects of adding liquefied sake lees to milk replacer on the growth performance, faecal characteristics, and blood metabolites of preweaning Japanese Black calves from 6 to 90days of age. Twenty-four Japanese Black calves at 6days of age were randomly assigned to one of three treatments: No liquefied sake lees (C, n=8), 100g/d (on a fresh matter basis) liquefied sake lees mixed with milk replacer (LS, n=8), and 200g/d (on a fresh matter basis) liquefied sake lees mixed with milk replacer (HS, n=8). The intake of milk replacer and calf starter, as well as, the average daily gain did not differ between the treatments. The number of days counted with faecal score 1 in LS was higher than in HS (P<0.05), while the number of days with diarrhoea medication in LS and C was lower than HS (P<0.05). The faecal n-butyric acid concentration tended to be higher in LS compared to C (P=0.060). The alpha diversity index (Chao1) was higher in HS than in C and LS at 90days of age (P<0.05). The principal coordinate analysis (PCoA) using weighted UniFrac distance showed that the bacterial community structures in faeces among the treatments at 90days of age were significantly different (P<0.05). The plasma β-hydroxybutyric acid concentration, an indicator of rumen development, was higher for LS than in C throughout the experiment (P<0.05). These results suggested that adding liquefied sake lees up to 100g/d (on a fresh matter basis) might promote rumen development in preweaning Japanese Black calves.

Metabolomic exploration of the effects of habituation to livestock trailer and extended transportation in goats.

Goats raised for meat production are often transported long distances. Twelve-month-old male Spanish goats were used to determine the effects of habituation to trailers on plasma metabolomic profiles when transported for extended periods. In a split-plot design, 168 goats were separated into two treatment (TRT; whole plot) groups and maintained on two different paddocks. Concentrate supplement was fed to one group inside two livestock trailers (habituated group, H), while the other group received the same quantity of concentrate, but not inside the trailers (non-habituated, NH). Goats were subjected to a 10-h transportation stress in 4 replicates (n = 21 goats/replicate/TRT) after 4 weeks of habituation period. Blood samples were collected prior to loading, 20min after loading (0h), and at 2, 4, 6, 8, and 10h of transportation (Time; subplot). A targeted quantitative metabolomics approach was employed to analyze the samples. The data were analyzed using R software and MIXED procedures in SAS. Several amino acids (alanine, serine, glycine, histidine, glutamate, trans-hydroxyproline, asparagine, threonine, methylhistidine, ornithine, proline, leucine, tryptophan) were higher (p < 0.05) in the H group compared to the NH group. Six long-chain acylcarnitines were higher (p < 0.05), while free (C0) and short-chain (C3, C5) carnitines were lower (p < 0.05) in the NH goats compared to the H goats. In general, amino acid concentrations decreased and long-chain acylcarnitine (>C10) levels increased with transportation time (p < 0.05). Butyric acid, α-ketoglutaric acid, and α-aminoadipic acid concentrations were lower (p < 0.05) and β-hydroxybutyric acid concentrations were higher in the NH goats compared to the H goats. Plasma glucose, non-esterified fatty acid (NEFA) and urea nitrogen concentrations were significantly influenced by Time (p < 0.01). Plasma NEFA concentrations were significantly lower (p < 0.01) in the H group than the NH group. Habituation to trailers can be beneficial in enhancing stress coping abilities in goats due to higher concentrations of metabolites such as butyrate and certain amino acids that support antioxidant activities and immune function. Plasma long-chain acylcarnitines may be good indicators of stress during long-distance transportation in goats.

Effects of Low or High Dosages of Dietary Sodium Butyrate on the Growth and Health of the Liver and Intestine of Largemouth Bass, Micropterus salmoides.

The concentration of butyric acid in the intestine increased with the increase in the content of fermentable dietary fibre; however, the potential physiological impact of a high dose of butyric acid on fish has not been sufficiently studied. The aim of this study was to investigate the effect of two dosages of butyric acid on the growth and health of the liver and intestine of the largemouth bass (Micropterus salmoides). Sodium butyrate (SB) was added to the diet at 0 g/kg (CON), 2 g/kg (SB2), and 20 g/kg (SB20), and the juvenile largemouth bass were fed to apparent satiation for 56 days. No significant difference was observed in the specific growth rate or hepatosomatic index among the groups (P > 0.05). The concentration of β-hydroxybutyric acid in the liver, the activities of alanine aminotransferase, aspartate aminotransferase, and alkaline phosphatase, and the concentrations of triglyceride and total cholesterol in serum increased significantly in the SB20 group compared to the CON group (P < 0.05). The relative expression of fas, acc, il1b, nfkb, and tnfa in the liver of the SB20 groups was also significantly higher than that of the CON group (P < 0.05). The above indicators in the group SB2 had similar change tendencies. The expression of nfkb and il1b in the intestine of both the SB2 and SB20 groups was significantly downregulated compared with that in the CON group (P < 0.05). The size of hepatocytes was enlarged, and the intracellular lipid droplets and the degree of hepatic fibrosis were increased in the SB20 group compared to the CON group. There was no significant difference in intestinal morphology among the groups. The above results indicated that neither 2 g/kg nor 20 g/kg SB had a positive effect on the growth of largemouth bass, while a high dosage of SB induced liver fat accumulation and fibrosis.

The Association between Blood Β-Hydroxybutyric Acid Concentration in the Second Week of Lactation and Reproduction Performance of Lithuanian Black and White Cows.

Simple SummaryDetermination of BHB concentration in the second week of lactation (WK 2) may allow us to predict the fertility properties of cows and help better manage farms. BHB concentration can be considered as a predictor trait of reproduction success. High BHB concentration requires a higher amount of insemination. The season in which the cows calve and the parity must be considered in the assessment as these factors affect BHB concentration in WK 2.Hyperketonemia is a very common metabolic state in dairy cows, which result in lower milk production, impaired fertility, and increased frequency of other diseases. In this study, we aimed to determine the influence of season, parity, and milk yield of cows on beta-hydroxybutyrate (BHB) concentration in the second week of lactation (WK 2) and establish the relationship between BHB concentration in WK 2 and reproduction performance traits such as insemination rate and first insemination day of Lithuanian Black and White dairy cows. The study included clinically healthy Lithuanian Black and White cows (n = 692). Blood BHB concentration was measured using capillary blood samples collected after morning milking when cows were 7–10 DIM. The impact of WK 2 blood BHB concentration on the insemination rate and first insemination day were investigated. The effect of BHB was evaluated according to the season, parity, and milk yield per lactation (305 DIM). Significant differences were observed in BHB concentration in WK 2 due to season and parity, but no statistically significant differences were observed for milk yields (305 d). Increased blood BHB concentration in WK 2 negatively affected insemination rate (p < 0.001) and first insemination day (p < 0.001). The study findings indicate that BHB concentration in WK 2 depends on season and parity, while the milk yield is not associated with BHB concentration. High BHB concentration in WK 2 increases insemination rate and delays the first insemination day for high milk-yielding Lithuanian Black and White dairy cows.

The relationship of asprosin with β-hydroxybutyric acid and postpartum disorders in cows.

The aim of the present study was to determine asprosin levels in cows, the relationship of this hormone with postpartum disorders and β-hydroxybutyric acid, and also the potential of asprosin to be a marker for postpartum diseases. The study was designed as a two-stage trial. In the first stage, blood asprosin and β-hydroxybutyric acid levels of 20 healthy Simmental cows aged 3-4 years were measured at the time of calving, and on days 3, 6, 9, 12 and 15 postpartum. In the second stage, 200 cows were divided into two groups: (1) healthy (n = 100) and (2) diseased (placental retention, hypocalcaemia, metritis, lameness, abomasal displacement, mastitis; n = 100); asprosin and β-hydroxybutyric acid concentrations of the blood were assessed on day 15 postpartum. In conclusion, the asprosin level was found to be at measurable levels in cows, and a negative correlation with β-hydroxybutyric acid was found. According to these findings, the data obtained from this study could be used for the prevention, control and treatment of some postpartum disorders associated with ketosis and for developing novel hypotheses concerning the actions of this hormone. It was concluded that further studies are required to reveal the associations between asprosin and postpartum disorders.

Dynamics of cholesterol fractions in cows during the transit period

This article describes the results of a study of cholesterol metabolism in cows during the transit period. Cows were selected with optimal and increased body fatness during the dry period three weeks before calving. Blood samples were taken twice before calving and twice after calving at regular intervals. In the blood serum, the concentration of cholesterol and its fractions - low-density lipoprotein cholesterol and high-density lipoprotein - was studied, as well as the concentration of β-hydroxybutyric acid on the fifth day after calving. Studies have shown unidirectional dynamics of cholesterol and its fractions during the observation period. Initially, a decrease in all indicators was determined, the minimum values were found during the first week after calving. By the end of the experiment, all cholesterol fractions increased. It was found that in cows with increased body condition during the dry period, the level of LDL cholesterol in the post-calving period was significantly lower by 19.0-25.4% than in cows with optimal body condition. It was also found that in well-fed cows ketogenesis is more active 5 days after calving. Thus, in cows with increased and optimal fatness, the concentration of β- hydroxybutyric acid was 1.63 ± 0.18 and 1.23 ± 0.11 mmol / l, respectively, which may indicate the presence of subclinical ketosis in obese cows.

Using the Relationship between Concentrations of Selected Whey Proteins and BHBA to Characterize the Metabolism of Dairy Cows in Early Lactation.

Simple SummaryA negative energy balance alters the concentration of various fractions of cows’ milk. Moreover, key changes are observed in the fat and protein content. Therefore, by analyzing the concentration of individual proteins in milk, it is possible to indirectly monitor the health of the animal. Subclinical ruminal acidosis and ketosis are the primary economic issues in dairy farming due to non-specific symptoms, difficulty in obtaining a diagnosis and reduced milk production. The aim of the present study was to identify the relationship between the concentrations of blood β-hydroxybutyric (BHBA) and whey proteins in milk as a marker for the diagnosis of metabolic diseases. Whey proteins were significantly influenced by both the lactation phase and BHBA. Therefore, it can be concluded that whey proteins can be used as non-invasive markers for diagnosing metabolic diseases. A high concentration of β-lactoglobulin can be a marker for diagnosing ketosis. Conversely, elevated levels of α-lactalbumin may indicate the occurrence of a metabolic disorder, such as acidosis.A negative energy balance alters the concentration of various fractions of cows’ milk. Therefore, by analyzing the concentration of individual proteins in milk, it is possible to indirectly monitor the health of the animal. The aim of the present study was to identify the relationship between the concentrations of blood β-hydroxybutyric acid (BHBA) and whey proteins in milk as a marker for the diagnosis of metabolic diseases. The analysis included milk and blood samples from 95 Holstein-Friesian cows, which were divided into three groups that were differentiated in terms of serum BHBA levels 5–7 days post-calving: LBHBA, low level of BHBA: 0.200–0.500 mmol/L; NBHBA, optimal level of BHBA- control group: 0.500–1.200 mmol/L; HBHBA, high level of BHBA: >1.200 mmol/L. Concentrations of α-lactoalbumin in the milk after 7 days of lactation proceeded in accordance with the concentration of β-hydroxybutyric acid, as follows: LBHBA > NBHBA > HBHBA. Concentrations of β-lactoglobulin in milk after 14 days of lactation proceeded in accordance with the concentration of β-hydroxybutyric acid, as follows: LBHBA < NBHBA < HBHBA. Therefore, it can be concluded that whey proteins can be used as non-invasive markers for diagnosing metabolic diseases. A high concentration of β-lactoglobulin can be a marker for diagnosing ketosis. Conversely, elevated levels of α-lactalbumin may indicate the occurrence of a metabolic disorder, such as acidosis.

Plasma metabolite changes in anestrous dairy cows with negative energy balance identified using 1H NMR technology

ABSTRACT The objective of the present study was to investigate the different plasma metabolites between anestrus and estrus postpartum dairy cows and to provide a theoretical basis for prevention of anestrus in dairy farm cows. In the experiment, one hundred and sixty-seven Holstein dairy cows were selected with similar age and parity. According to the concentration of β-hydroxybutyric acid, non-esterified fatty acids and glucose in plasma during 14 to 21 days in milk, all dairy cows were determined as having a status of energy balance. According to the results of clinical symptom, rectal and B ultrasound examination at 60 to 90 days postpartum, these cows were divided into twenty estrus and twenty-four anestrus group, other dairy cows were removed. 1H nuclear magnetic resonance technology was utilized to detect the plasma metabolites changes and screen different plasma metabolites between anestrus and estrus cows. Ten different metabolites including alanine, glutamic acid, asparagine, creatine, choline, phosphocholine, glycerophosphocholine, low-density lipoprotein, and very-low-density lipoprotein were significantly decreased in anestrous cows compared with estrous cows. Metabolic pathway analyses indicated that differential metabolites were primarily involved in amino acid and glycerophospholipid metabolism. These metabolites and their enrichment pathways indicate that reduced steroid hormone synthesis precursors result in lower levels of estradiol and progesterone and cause anestrus in negative energy balance. These data provide a better understanding of the changes that may affect estrus of postpartum dairy cows at NEB status and lay the ground for further research.

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

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

Adding monoglycerides containing short and medium chain fatty acids to milk replacer: effects on health and performance of preweaned calves

The aim of this trial was to investigate the effects of a blend of monoglycerides of short- and medium-chain fatty acids (SMCFA) on health status and growth of preweaned male calves from 16 to 72 days of age. The in vivo study was conducted as a complete randomised block design. Twenty Italian Holstein calves received starter administered ad libitum and 6 L of milk replacer/day (12% powder concentration) and were divided into treated (T) and control (C) groups. A preliminary in vitro antibiotic activity test of SMCFA was performed on several bacterial strains showing a minimum inhibiting concentration ranging from 0.8 to 12.5%. The group T received SMCFA (11 g/day) within the milk replacer while, to obtain isoenergetic liquid diets, the group C received additional 18 g/day of milk replacer powder. Body weight and morphological measurements of the calves were performed weekly. Health scores were checked daily. Faecal and blood samples were collected every two weeks for routine clinical investigations including serum concentration of non-esterified fatty acids and β-Hydroxybutyric acid. At the end of the trial calves were slaughtered and gastrointestinal tract (GIT) organs weighed. The group T reported a better health status and did not require Ab therapy. No differences in biometrical measurements, feed efficiency, blood parameters, and GIT measures were found between the two groups. At 44 days of age, group T showed higher β-Hydroxybutyric acid concentration compared to group C (p < .05). The SMCFA improved the calf health status avoiding the use of antimicrobics. HIGHLIGHTS The monoglycerides of short and medium chain fatty acids improve calf health status. The monoglycerides of short and medium chain fatty acids reduce the need for antibiotic treatment. The monoglycerides of short and medium chain fatty acids reduce the negative impact of enteropathogens.

Dietary Supplementation of a Live Yeast Product on Dairy Sheep Milk Performance, Oxidative and Immune Status in Peripartum Period.

This study evaluated the dietary administration of Saccharomyces cerevisiae live yeast on milk performance and composition, oxidative status of both blood plasma and milk, and gene expression related to the immune system of lactating ewes during the peripartum period. Chios ewes were fed either a basal diet (BD) (Control, n = 51) or the BD supplemented with 2 g of a live yeast product/animal (ActiSaf, n = 53) from 6 weeks prepartum to 6 weeks postpartum. Fatty acid profile, oxidative, and immune status were assessed in eight ewes per treatment at 3 and 6 weeks postpartum. The β-hydroxybutyric acid concentration in blood of ActiSaf fed ewes was significantly lower in both pre- and postpartum periods. A numerical increase was found for the milk yield, fat 6% corrected milk (Fat corrected milk (FCM6%)), and energy corrected milk yield (ECM) in ActiSaf fed ewes, while daily milk fat production tended to increase. The proportions of C15:0, C16:1, C18:2n6t, and C18:3n3 fatty acids were increased in milk of ActiSaf fed ewes, while C18:0 was decreased. Glutathione reductase in blood plasma was increased (p = 0.004) in ActiSaf fed ewes, while total antioxidant capacity measured by 2,2′-Azino-bis (3-ethylbenzthiazoline-6-sulfonic acid) (ABTS) method was decreased (p < 0.001). Higher ABTS values were found in the milk of the treated group. The relative transcript levels of CCL5, CXCL16, and IL8 were suppressed, while that of IL1B tended to decrease (p = 0.087) in monocytes of ActiSaf fed ewes. In conclusion, the dietary supplementation of ewes with S. cerevisiae, improved the energy utilization and tended to enhance milk performance with simultaneous suppression on mRNA levels of pro-inflammatory genes during the peripartum period.

Metabolomics reveals effects of rumen-protected glucose on metabolism of dairy cows in early lactation

Early lactation dairy cows are vulnerable to negative energy balance (NEB) due to reduced dry matter intake (DMI) and high energy requirements for lactation. This study mainly revealed the mechanism of different doses of rumen-protected glucose (RPG) to alleviate NEB of the early lactating dairy cows through serum untargeted metabolomics, and explored the effects of different doses of RPG on rumen fermentation, lactation performance, and serum biochemical indices associated with NEB. Thirty-two multiparous Holstein cows in early lactation, blocked by parity, liveweight, and milk yield, were allocated to one of four treatments in a randomized block design. Treatments were: control (CON), low RPG (LRPG), medium RPG (MRPG) and high RPG (HRPG) with 0, 200, 350 and 500 g RPG per cow per day, respectively. Treatments were applied from calving to 35 days postpartum. The results showed that compared with other groups, the MRPG group changed the rumen environment, increased milk protein composition on 35 d, and decreased serum concentrations of β-hydroxybutyric acid (BHBA), non-esterified fatty acid (NEFA), aspartate aminotransferase (AST), and could increase glucose (GLU) concentration in serum. Dietary RPG supplementation affected ruminal pH and some volatile fatty acid (VFA) concentrations. The results of serum metabolomics showed that the metabolites with the largest contribution of the first 30 differences after RPG supplementation were mainly involved in lipid metabolism. The results of this study indicated that adding 350 g RPG to a cow’s diet in early lactation could improve energy balance by reducing fat mobilization.

The effect of glycerol or rumen-protected choline chloride on rumen fermentation and blood metabolome in pregnant ewes suffering from negative energy balance

The objective of this study was to investigate the counteracting effects of glycerol or rumen-protected choline chloride supplementation on rumen fermentation and the profile changes in blood metabolites in pregnant ewes suffering from negative energy balance. Eighteen pregnant Hu sheep with a gestation period of 100 days were used in the present study. Feed restriction was started from 100 days of gestation and lasted for 9 days. During the period of feed restriction, these sheep were randomly assigned to three groups: the control group (CON, n = 6), which received no supplementation, the glycerol group (GLY, n = 6), which received 40 mL of glycerol per day, and the rumen-protected choline chloride group (RPC, n = 6), which received 10 g of rumen-protected choline chloride per day lasted 9 days. All supplements were mixed with feed. The results showed that, compared to CON, glycerol supplementation decreased the proportion of acetate (P < 0.01) and increased the proportion of propionate (P < 0.01). The proportions of butyrate (P = 0.04) and valerate (P = 0.04) were higher in GLY than in RPC, but there was no significant difference in their proportions between GLY and CON. Blood β-hydroxybutyric acid concentration continuously increased in CON during the treatment period, while it remained stable in the ewes receiving glycerol or rumen-protected choline chloride. Glucose levels in the blood of ewes receiving glycerol or rumen-protected choline chloride were higher than those in CON (P = 0.01). Compared to CON, the levels of non-esterified fatty acids (NEFA) (P = 0.02) and very-low-density lipoproteins (VLDL) (P = 0.04) in the blood were increased in ewes that received rumen-protected choline chloride. Correlation analysis of the DEMs and rumen fermentation parameters showed a positive correlation between citric acid and phosphohydroxypyruvic acid (r = 0.61, P < 0.01), as well as between bilirubin and BHBA (r = 0.59, P < 0.01). d-Glucose have a positive correlation with propionate (r = 0.48, P = 0.02), while have a negative correlation with BHBA (r = −0.77, P < 0.01), 2-hydroxylauroylcarnitine (r = -0.53, P < 0.01), 2-octenoylcarnitine (r = −0.61, P < 0.01), and 3-hydroxydodecanoic acid (r = −0.60, P = 0.04). It is concluded that the supplementation of glycerol or rumen-protected choline chloride counteracted changes in metabolite concentrations in ewes that had been induced by feed restriction.

Sorghum silage substituted for corn silage in diets for dairy cows: Effects on feed intake, milk yield and quality, and serum metabolites

ABSTRACT Objective Whole-plant corn silage (CS) is the main forage fed to dairy cows in China, usually comprising about 35% of the total DM of the TMR. However, sorghum is more resistant to drought and tolerates soils that are less fertile compared with corn, and consequently, sorghum silage (SS) would be preferable to CS in many drought-prone areas of China. Sweet SS is characterized by a high content of water-soluble carbohydrates and energy yield. Previous studies examined the effects of feeding either CS or SS in the TMR of lactating cows. The aim of the present study was to assess the effects of substituting different levels of sweet SS for CS in the TMR fed to dairy cows in a smallholder farm in western China. Materials and Methods Sixteen multiparous Holstein cows (mean BW of 454 ± 25 kg), producing approximately 12 kg/d milk, were assigned randomly to 4 dietary treatments. Experimental diets were formulated to be iso-energetic and iso-nitrogenous with 45% forage and 55% concentrate, and the diets included 10% oat hay and variable proportions of CS and SS. Dietary treatments included (1) 100% CS (35% DM of TMR, control diet); (2) 35% of CS replaced by SS; (3) 65% of CS replaced by SS; and (4) 100% of CS replaced by SS. Results and Discussion The DMI decreased linearly (P = 0.002) as SS increased. Milk yield and lactose percentage did not differ among treatments, but lactose yield exhibited a cubic response (P = 0.004), whereas milk fat percentage tended to increase linearly (P = 0.077) as SS increased. Increasing SS resulted in a quadratic response in milk protein percentage and yield (P = 0.001) and MUN increased linearly as SS increased (P = 0.022). The ECM/DMI ranged between 0.75 and 0.82 and increased linearly as SS increased (P = 0.029). Although there were differences among treatments in serum concentrations of aspartate transaminase, alanine aminotransferase, alkaline phosphatase, urea nitrogen, and glucose, all concentrations were in the normal range for healthy, lactating cows. Serum β-hydroxybutyric acid concentrations were well below the subclinical levels in all treatments. The SCC decreased linearly (P = 0.024) with increasing SS. Implications and Applications We concluded that for low-producing dairy cows, SS can replace CS in total in the diet without adverse effects on milk yield and its components and on serum metabolites. This is particularly important for small dairy herds in dry areas.