Transcriptional modulation of heat shock proteins and adipogenic regulators in bovine adipocytes following heat exposure.

This research endeavored to elucidate the transcriptional modulation of heat shock proteins and adipogenic regulators in bovine subcutaneous adipocytes following thermal exposure. Post-differentiation, mature adipocytes were subjected to three treatments of control (CON), moderate (MHS), and extreme (EHS) heat stress. These treatments consist of thermal conditions at temperatures of 38 °C (CON), 39.5 °C (MHS), or 41 °C (EHS) along with of 3 or 12 h. There was no statistically significant variations observed in the gene expressions of HSP27 and HSP70 when comparing CON with MHS across both exposures. Contrastingly, when comparing CON with EHS, an upregulation (P < 0.01) in HSP27 gene expression was evident for both 3 and 12 h of incubation, while HSP70 gene expression exhibited elevation (P < 0.01) at the 3-h mark, with no change observed at 12 h. Protein quantification, however, revealed an elevation (P < 0.01) in HSP27 and HSP70 for both CON vs. MHS and CON vs. EHS at the 12-h exposure. This trend in protein level mirrored (P < 0.05) that of proliferator-activated receptor-gamma (PPARγ). Elevated (P < 0.05) protein levels of fatty acid synthase (FAS) were exclusively discernible in the CON vs. MHS. Increased (P < 0.01) transcriptional activity of PPARγ, CCAAT/enhancer-binding protein alpha (C/EBPα), stearoyl-CoA desaturase (SCD), and FAS was evident in the CON vs. EHS comparison. Complementary to these molecular findings, an augmented lipid droplet accumulation was observed (P < 0.01) in EHS-exposed adipocytes progressively from day 6 through day 9. Our current study highlights how different levels and lengths of heat stress can impact the activity of important heat-related proteins and factors that play a role in fat development in beef cattle. These findings can help guide strategies to manage how beef cattle are exposed to heat, which can affect fat storage and ultimately the quality of the meat's marbling.

Effects of Rumen-Protected L-Tryptophan Supplementation on Productivity, Physiological Indicators, Blood Profiles, and Heat Shock Protein Gene Expression in Lactating Holstein Cows under Heat Stress Conditions.

In this study, we examined the effects of rumen-protected L-tryptophan supplementation on the productivity and physiological metabolic indicators in lactating Holstein cows under heat stress conditions. The study involved eight early lactating Holstein cows (days in milk = 40 ± 9 days; milk yield 30 ± 1.5 kg/day; parity 1.09 ± 0.05, p < 0.05), four cows per experiment, with environmentally controlled chambers. In each experiment, two distinct heat stress conditions were created: a low-temperature and low-humidity (LTLH) condition at 25 °C with 35-50% humidity and a high-temperature and high-humidity (HTHH) condition at 31 °C with 80-95% humidity. During the adaptation phase, the cows were subjected to LTLH and HTHH conditions for 3 days. This was followed by a 4-day heat stress phase and then by a 7-day phase of heat stress, which were complemented by supplementation with rumen-protected L-tryptophan (ACT). The findings revealed that supplementation with ACT increased dry matter intake as well as milk yield and protein and decreased water intake, heart rate, and rectal temperature in the HTHH group (p < 0.05). For plateletcrit (PCT, p = 0.0600), the eosinophil percentage (EOS, p = 0.0880) showed a tendency to be lower, while the monocyte (MONO) and large unstained cells (LUC) amounts were increased in both groups (p < 0.05). Albumin and glucose levels were lower in the HTHH group (p < 0.05). The gene expressions of heat shock proteins 70 and 90 in the peripheral blood mononuclear cells were higher in the ACT group (HTHH, p < 0.05). These results suggest that ACT supplementation improved productivity, physiological indicators, blood characteristics, and gene expression in the peripheral blood mononuclear cells of early lactating Holstein cows under heat-stress conditions. In particular, ACT supplementation objectively relieved stress in these animals, suggesting that L-tryptophan has potential as a viable solution for combating heat-stress-induced effects on the cattle in dairy farming.

Temperature Fluctuations Modulate Molecular Mechanisms in Skeletal Muscle and Influence Growth Potential in Beef Steers.

Our investigation elucidated the effects of severe temperature fluctuations on cellular and physiological responses in beef cattle. Eighteen Red Angus beef steers with an average body weight of 351 ±â€…24.5 kg were divided into three treatment groups: 1) Control (CON), exposed to a temperature-humidity index (THI) of 42 for 6 h without any temperature changes; 2) Transport (TP), subjected to a one-mile trailer trip with a THI of 42 for 6 h; and 3) Temperature swing (TS), exposed to a one-mile trailer trip with a THI shift from 42 to 72-75 for 3 h. Our findings indicate that TS can induce thermal stress in cattle, regardless of whether the overall temperature level is excessively high or not. Behavioral indications of extreme heat stress in the cattle were observed, including extended tongue protrusion, reduced appetite, excessive salivation, and increased respiratory rate. Furthermore, we observed a pronounced overexpression (P < 0.05) of heat shock proteins (HSPs) 20, 27, and 90 in response to the TS treatment in the longissimus muscle (LM). Alterations in signaling pathways associated with skeletal muscle growth were noted, including the upregulation (P < 0.01) of Pax7, Myf5, and myosin heavy chain (MHC) isoforms. In addition, an increase (P < 0.05) in transcription factors associated with adipogenesis was detected (P < 0.05), such as PPARγ, C/EBPα, FAS, and SCD in the TS group, suggesting the potential for adipose tissue accumulation due to temperature fluctuations. Our data illustrated the potential impacts of these temperature fluctuations on the growth of skeletal muscle and adipose tissue in beef cattle.

In this study, we investigated the effects of severe temperature fluctuations on beef cattle and their cellular and physiological responses. Our findings demonstrate that even moderate temperature swings can cause thermal stress in cattle, leading to observable behavioral signs such as extended tongue protrusion, reduced appetite, excessive salivation, and increased respiratory rate. We also observed a significant increase in the expression of heat shock proteins (HSPs), which protect cells from stress, indicating their importance as early responders to temperature fluctuations. Furthermore, we examined the signaling pathways involved in skeletal muscle growth and found that severe temperature fluctuations can stimulate the upregulation of myogenic regulatory factors and myosin heavy chains. These changes suggest an increased demand for muscle contractile properties and hyperplasia during temperature challenges. In addition, our study revealed alterations in transcription factors associated with adipogenesis, such as PPARγ and C/EBPα, indicating the potential for adipose tissue accumulation in response to temperature fluctuations.

Heat shock protein 27 regulates myogenic and self-renewal potential of bovine satellite cells under heat stress.

While satellite cells play a key role in the hypertrophy, repair, and regeneration of skeletal muscles, their response to heat exposure remains poorly understood, particularly in beef cattle. This study aimed to investigate the changes in the transcriptome, proteome, and proliferation capability of bovine satellite cells in response to different levels of heat stress (HS) and exposure times. Satellite cells were isolated from 3-mo-old Holstein bulls (body weight: 77.10 ± 2.02 kg) and subjected to incubation under various temperature conditions: 1) control (38 °C; CON), 2) moderate (39.5 °C; MHS), and extreme (41 °C; EHS) for different durations ranging from 0 to 48 h. Following 3 h of exposure to extreme heat (EHS), satellite cells exhibited significantly increased gene expression and protein abundance of heat shock proteins (HSPs; HSP70, HSP90, HSP20) and paired box gene 7 (Pax7; P < 0.05). HSP27 expression peaked at 3 h of EHS and remained elevated until 24 h of exposure (P < 0.05). In contrast, the expression of myogenic factor 5 (Myf5) and paired box gene 3 (Pax3) was decreased by EHS compared to the control at 3 h of exposure (P < 0.05). Notably, the introduction of HSP27 small interference RNA (siRNA) transfection restored Myf5 expression to control levels, suggesting an association between HSP27 and Myf5 in regulating the self-renewal properties of satellite cells upon heat exposure. Immunoprecipitation experiments further confirmed the direct binding of HSP27 to Myf5, supporting its role as a molecular chaperone for Myf5. Protein-protein docking algorithms predicted a high probability of HSP27-Myf5 interaction as well. These findings indicate that extreme heat exposure intrinsically promotes the accumulation of HSPs and modulates the early myogenic regulatory factors in satellite cells. Moreover, HSP27 acts as a molecular chaperone by binding to Myf5, thereby regulating the division or differentiation of satellite cells in response to HS. The results of this study provide a better understanding of muscle physiology in heat-stressed cells, while unraveling the intricate molecular mechanisms that underlie the HS response in satellite cells.

This study aimed to elucidate the response of bovine satellite cells to heat exposure. Satellite cells were isolated from Holstein bulls and subjected to varying temperatures. Transcriptional, proteomic, and proliferative changes were assessed. Following extreme heat exposure, cells exhibited upregulated expression of heat shock proteins (HSPs; HSP70, HSP90, HSP20) and paired box gene 7 (Pax7). Conversely, the expression of myogenic factor 5 (Myf5) and paired box gene 3 (Pax3), key regulators of myogenesis, decreased under conditions of extreme heat. Notably, downregulation of HSP27 expression using siRNA restored Myf5 expression to normal levels, implying an association between HSP27 and Myf5 in the modulation of satellite cell properties during heat exposure. Our results validated the direct binding of HSP27 to Myf5, substantiating its role as a molecular chaperone. These findings underscore the elevation of HSPs, and alteration of early myogenic regulatory factors implicated in muscle development upon exposure to extreme heat. HSP27 functions as a molecular chaperone by engaging with Myf5, thereby influencing the division or differentiation of satellite cells during heat stress (HS). This study contributes to the advancement of our comprehension regarding the muscular physiology of heat-stressed animals, while clarifying the intricate molecular mechanisms governing the response of satellite cells to HS.

Effects of different energy levels and two levels of temperature-humidity indices on growth, blood metabolites, and stress biomarkers in Korean native calves.

This study investigated the effects of dietary energy levels on growth, blood metabolites, and stress biomarkers in Korean native calves subjected to heat stress (HS). Twenty-four calves (BW: 221.5 ± 24.9 kg; age: 162 ± 4.8 d) were randomly housed in climate-controlled chambers using 3 × 2 factorial design. There were three treatment groups including low energy (LE = 2.53), medium energy (ME = 2.63), and high energy levels (HE = 2.72 Mcal/kg of DM) and two stress levels (threshold: THI = 70-73; severe: THI = 89-91). The calves were adapted to 22 °C for 7 days, then to the target THI level for 14 days. Energy intake, average daily gain, and gain to feed ratio were determined to decline (p < 0.05) under severe HS compared with threshold. Under severe HS, rectal temperature was increased 0.67 °C compared with threshold. Severe HS increased glycine, ammonia, and 3-methylhistidine concentrations compared with threshold (p < 0.05). Gluconeogenic AAs in the blood were increased among the various energy levels regardless of HS. In PBMCs the expression of HSP70 gene was increased in the LE group (p < 0.05), and the HSP90 gene expression was increased in LE and ME groups (p < 0.05) under severe HS. However, the expression of genes HSP70 and HSP90 in HE group did not differ under severe HS (p > 0.05). It has been suggested that HE intake may have a beneficial effect on PBMCs by mitigating ATP depletion. No differences in growth performance were found when increasing energy intake with high protein (CP 17.5%) under HS. However, the increase in energy levels resulted in increased gluconeogenic AAs but decreased urea and 3-methylhistidine in blood. In conclusion, increased energy levels are thought to improve HS adaptability by inhibiting muscle degradation and glucose production using gluconeogenic AAs in Korea native calves under HS condition.

Exploring the impact of temporal heat stress on skeletal muscle hypertrophy in bovine myocytes.

The primary aim of this investigation was to explore the impact of different temporal stress conditions on the regulators associated with skeletal muscle hypertrophy in bovine myocytes. Bovine satellite cells (BSCs) were extracted from three-month-old Holstein bull calves and subjected to myogenic differentiation under three thermal treatments: 38 °C (control; CON), 39.5 °C (moderate heat stress; MHS), and 41 °C (extreme heat stress; EHS) for a duration of 3 or 48 h. Exposure to EHS resulted in elevated (P < 0.01) expression levels of heat shock protein (HSP)20, HSP27, HSP70, and HSP90, along with increased (P < 0.01) protein levels. Moreover, cells exposed to MHS and EHS exhibited enhanced (P < 0.01) gene expression of myoblast determination protein 1 (MyoD), while myogenin (MyoG) was overexpressed (P < 0.01) in cells exposed to EHS. These findings suggest that heat exposure can potentially induce myogenic differentiation through the modulation of myogenic regulatory factors. Furthermore, our investigations revealed that exposure to EHS upregulated (P < 0.01) myosin heavy chain (MHC) I expression, whereas MHC IIA (P < 0.01) and IIX (P < 0.01) expression were increased; P < 0.01) under MHS conditions. These observations suggest that the temperature of the muscle may alter the proportion of muscle fiber types. Additionally, our data indicated that EHS activated (P < 0.01) the expression of insulin-like growth factor 1 (IGF-1) and triggered the activation of the Akt/mTOR/S6KB1 pathway, a known anabolic pathway associated with cellular protein synthesis. Consequently, these altered signaling pathways contributed to enhanced protein synthesis and increased myotube size. Overall, the results obtained from our current study revealed that extreme heat exposure (41 °C) may promote skeletal muscle hypertrophy by regulating myogenic regulatory factors and IGF-1-mediated mTOR pathway in bovine myocytes.

Heat Stress Effects on Physiological and Blood Parameters, and Behavior in Early Fattening Stage of Beef Steers.

This study was conducted to investigate the effect of heat stress (HS) on physiological, blood, and behavioral parameters, according to the temperature-humidity index (THI), in beef steers. Twelve Korean native beef steers (342.7 ± 13.81 days old, body weight (BW) of 333.0 ± 18.53 kg) were used in this experiment. Beef steers were randomly distributed into three homogenized groups (four beef steers each) for 14 days, namely, threshold (THI = 64-71), mild-moderate (THI = 72-79), and severe (THI = 80-87). Feed and water intake were recorded daily. Physiological parameters, including heart rate and rectal temperature, and behavioral patterns (standing and lying down) were measured weekly. Blood was sampled every week to analyze hormones, heat shock protein (HSP) levels, metabolites, and hematological parameters. All data were analyzed using repeated-measures analysis. Beef steers exposed to severe THI had significantly increased (p < 0.001) water intake, heart rate, and rectal temperature compared to the threshold and mild-moderate THI beef steers. Additionally, increased blood cortisol (p < 0.001), HSP70 (p < 0.001), blood urea nitrogen (BUN) (p = 0.014), and time spent standing (p < 0.001) were observed in beef steers after exposure to severe THI compared to beef steers in the threshold and mild-moderate THI groups. However, dry matter intake, blood glucose, and non-esterified fatty acids were not different among the THI groups. In conclusion, heart rate, rectal temperature, blood cortisol, HSP70, BUN, and time spent standing were closely associated with severe HS conditions in beef steers. These phenomena indicated that beef steers exposed to HS modulated their behavior and blood parameters, as well as their physiological response, to maintain homeostasis.

Impact of Cold Stress on Physiological, Endocrinological, Immunological, Metabolic, and Behavioral Changes of Beef Cattle at Different Stages of Growth.

The purpose of this study was to investigate the effect of cold stress (CS) on the physiological, blood, and behavioral parameters of beef cattle according to their growth stage. Twelve calves in the growing stages (220.4 ± 12.33 kg, male and non-castrated) and twelve steers in the early fattening stages (314.2 ± 18.44 kg) were used in this experiment. The animals were randomly distributed into three homogenized groups (four animals each) for 14 days, namely threshold, mild-moderate cold stress (MCS), and extreme cold stress (ECS), according to the outside ambient temperature. The feed and water intakes were recorded daily. The physiological parameters, blood parameters, and behavioral patterns were measured weekly. All data were analyzed using repeated-measures analysis. The calves exposed to the ECS decreased (p < 0.064, tendency) their dry matter intake compared to the threshold and MCS groups. The HR and RT increased (p < 0.001) in the ECS compared to the threshold in calves and steers. Moreover, increased (p < 0.05) blood cortisol, non-esterified fatty acids (NEFA), and time spent standing were observed after exposure to ECS in calves and steers. However, the calves exposed to the ECS had decreased (p = 0.018) blood glucose levels compared to the threshold. In conclusion, ECS affects the dry matter intake, HR, RT, blood cortisol, NEFA, and behavioral patterns in beef calves and steers. This phenomenon indicated that beef cattle exposed to CS modulated their behavior and blood parameters as well as their physiological response to maintain homeostasis regardless of the growth stage.

Identification of Potential Biomarkers and Metabolic Pathways of Different Levels of Heat Stress in Beef Calves.

Heat stress (HS) damages the global beef industry by reducing growth performance causing high economic losses each year. However, understanding the physiological mechanisms of HS in Hanwoo calves remains elusive. The objective of this study was to identify the potential biomarkers and metabolic pathways involving different levels of heat stress in Hanwoo calves. Data were collected from sixteen Hanwoo bull calves (169.6 ± 4.6 days old, BW of 136.9 ± 6.2 kg), which were maintained at four designated ranges of HS according to the temperature−humidity index (THI) including: threshold (22 to 24 °C, 60%; THI = 70 to 73), mild (26 to 28 °C, 60%; THI = 74 to 76), moderate (29 to 31 °C, 80%; THI = 81 to 83), and severe (32 to 34 °C, 80%; THI = 89 to 91) using climate-controlled chambers. Blood was collected once every three days to analyze metabolomics. Metabolic changes in the serum of calves were measured using GC-TOF-MS, and the obtained data were calculated by multivariate statistical analysis. Five metabolic parameters were upregulated and seven metabolic parameters were downregulated in the high THI level compared with the threshold (p < 0.05). Among the parameters, carbohydrates (ribose, myo-inositol, galactose, and lactose), organic compounds (acetic acid, urea, and butenedioic acid), fatty acid (oleic acid), and amino acids (asparagine and lysine) were remarkably influenced by HS. These novel findings support further in-depth research to elucidate the blood-based changes in metabolic pathways in heat-stressed Hanwoo beef calves at different levels of THI. In conclusion, these results indicate that metabolic parameters may act as biomarkers to explain the HS effects in Hanwoo calves.

Evaluation of the Feed Nutritional Value of Noni (Morinda citrifolia) Meal for Holstein Dairy Cows.

In three consecutive studies, we evaluated the effects of noni (Morinda citrifolia) meal on rumen fermentation and degradation characteristics, production performance, physiological parameters, and milk fatty acid profile in Holstein dairy cows. In in vitro (first study) and in situ (second study) experiments, rumen fluids from two fistulated Holstein dairy cows were used. The concentration of noni meal added was 0 (control), 1, 3, 5, or 7% of the basal diet (DM basis). In the in situ experiment, wheat bran was used as a control. Triplicated bags were incubated for 0, 4, 8, 12, 24, 48, 72, or 96 h. In an in vivo experiment (third study), 38 Holstein cows (145 ± 87 days DIM; 1.8 ± 0.9 parity; 35.4 ± 6.3 kg/day milk yield) were equally assigned to the control and treatment groups (19 cows each). Basal feed and noni meal pellets (1.5% of total feed DM basis) were fed to the treatment group. The control group was also fed the basal feed and pellets containing 0% noni meal. There were no significant differences in in vitro dry matter digestibility, pH, total gas production (TGP), CH4, NH3-N, and volatile fatty acids (p > 0.05). In the in situ experiments, the crude protein (CP) rapidly soluble fraction 'a' (CP-a) was higher in noni meal than in wheat bran, and rumen degradable protein was also higher in noni meal than in wheat bran. In the in vivo experiments, when noni meal pellets were fed, there was no significant difference in milk yield and composition, but the triglyceride levels decreased (p < 0.05), the C18:1 fatty acid level increased (p < 0.05), and the C18:0 fatty acid level decreased (p < 0.05). Collectively, noni meal can be used as a feed ingredient up to 1.5% (total feed DM basis) in Holstein dairy cows and as feed supplementation to increase the C18:1 fatty acid level in milk.

Effects of different protein levels on growth performance and stress parameters in beef calves under heat stress.

This study investigated the effects of dietary protein levels under various heat stress (HS) conditions on the growth performance and stress parameters in Korean native beef calves. Male calves (n = 40; initial BW = 202.2 ± 3.31 kg) were randomly assigned to climatic-controlled chambers with 3 × 3 factorial arrangements. Calves were assigned into three dietary protein levels (low protein; LP = 12.5%, medium protein; MP = 15%, and high protein; HP = 17.5%) and three HS levels [mild: temperature-humidity index (THI) = 74 to 76, moderate: THI = 81 to 83, and severe: THI = 89 to 91] with control (threshold: THI = 70 to 73 and dietary protein level 12.5%). The calves were subjected to ambient temperature (22 °C) for 7 days and subsequently to the temperature and humidity corresponding to the target THI level for 21 days. The data were analyzed using the repeated-measures analysis by the GLM procedure of SAS. As a result, average daily gain (ADG) was decreased (P < 0.05) under severe HS level compared to the mild and moderate HS stress levels. However, HP increased ADG (P < 0.05) than moderate levels (LP) and severe levels (LP and MP). Under different HS levels (mild, moderate, and severe), HR, RT, and blood cortisol were increased (P < 0.05) compared to a threshold level, but no differences were observed in the parameters among various protein levels. Varied HS levels decreased the levels of blood glucose, NEFA, and amino acids (AAs) (lysine and glutamic acid) compared to a threshold (P < 0.05). But, the HP group resulted in increased (P < 0.05) levels of blood glucose, NEFA, and AAs (lysine and glutamic acid) compared to LP and MP groups under severe HS stress. The expression level of the HSP70 gene in peripheral blood mononuclear cell (PBMC) and hair follicles was increased (P < 0.05) following an increase in moderate and severe HS levels. Also, HSP70 gene expression in the HP group was decreased (P < 0.05) compared with LP and MP groups under intense HS level. Overall, HS in Korean native beef calves exhibited negative effects on ADG, blood glucose, NEFA, and AA profile. However, 17.5% of dietary protein (HP) could compensate for the growth of heat-exposed Korean native beef calves through the regulation of homeostasis by protein and energy metabolism. Also, it was evident that adequate protein (HP) is used as a major nutrient for HSP70 synthesis in PBMC and hair follicles causing, a boost in the immune system of heat-exposed Korean native beef calves.

Dietary supplementation with L-glutamine enhances immunity and reduces heat stress in Hanwoo steers under heat stress conditions.

This study investigated the effects of L-glutamine (Gln) supplementation on growth performance, physiological traits, heat shock proteins (HSPs), and gene expression related to muscle and adipose tissue development in Hanwoo steers under heat stress (HS) conditions. Eight Hanwoo steers (initial body weight [BW] 570.7 ± 43.6 kg, months of age 22.3 ± 0.88) were randomly separated into two groups, control and treatment, and supplied with the concentration (1.5% of BW kg/day/head) and rice straw (1.5 kg/day/head). The treatment group were fed the Gln supplementation (0.5% of concentration, as-fed basis) once a day at 08:00 h. Blood samples for the assessment of haematological and biochemical parameters and the separation of peripheral blood mononuclear cells (PBMCs) were collected four times, at 0, 3, 6, and 10 weeks of the experiment. Feed intake was measured daily. BW to analyze growth performance and hair follicle collection to analyze the expression of HSPs were executed four times at 0, 3, 6, and 10 weeks. To analyze gene expression, longissimus dorsi muscle samples were collected by biopsy at the end of the study. As a result, growing performance, including final BW, average daily gain, and gain-to-feed ratio, were not different between the two groups. Leukocytes including lymphocytes and granulocytes, tended to increase in the Gln supplementation group (p = 0.058). There were also no differences in biochemical parameters shown between the two groups, except total protein and albumin, both of which were lower in the Gln supplementation group (p < 0.05). Gene expressions related to muscle and adipose tissue development were not different between the two groups. As temperature-humidity index (THI) increased, HSP70 and HSP90 expression in the hair follicle showed a high correlation. HSP90 in the hair follicle was decreased in the treatment group compared with the control group at 10 weeks (p < 0.05). Collectively, dietary Gln supplementation (0.5% of concentration, as-fed basis) may not be influential enough to affect growth performance and gene expression related to muscle and adipose tissue development in steers. However, Gln supplementation increased the number of immune cells and decreased HSP90 in the hair follicle implying HS reduction in the corresponding group.

Chemerin Regulates Epithelial Barrier Function of Mammary Glands in Dairy Cows.

Epithelial barrier function in the mammary gland acts as a forefront of the defense mechanism against mastitis, which is widespread and a major disorder in dairy production. Chemerin is a chemoattractant protein with potent antimicrobial ability, but its role in the mammary gland remains unelucidated. The aim of this study was to determine the function of chemerin in mammary epithelial tissue of dairy cows in lactation or dry-off periods. Mammary epithelial cells produced chemerin protein, and secreted chemerin was detected in milk samples. Chemerin treatment promoted the proliferation of cultured bovine mammary epithelial cells and protected the integrity of the epithelial cell layer from hydrogen peroxide (H2O2)-induced damage. Meanwhile, chemerin levels were higher in mammary tissue with mastitis. Tumor necrosis factor alpha (TNF-α) strongly upregulated the expression of the chemerin-coding gene (RARRES2) in mammary epithelial cells. Therefore, chemerin was suggested to support mammary epithelial cell growth and epithelial barrier function and to be regulated by inflammatory stimuli. Our results may indicate chemerin as a novel therapeutic target for diseases in the bovine mammary gland.

Responses of beef calves to long-term heat stress exposure by evaluating growth performance, physiological, blood and behavioral parameters.

The objective of this study was to explore the responses of beef calves to long-term heat stress (HS) exposure at various levels in comparison with the animals under thermoneutral conditions by evaluating growth performance, physiological, blood, and behavioural parameters. Data were collected from sixteen beef calves (BW: 136.9 ± 6.23 kg; age: 169.6 ± 4.60 d) kept at four stress levels of designated temperature humidity index (THI): threshold (22-24 °C, 60%; THI = 70 to 73), mild (26-28 °C, 60%; THI = 74 to 76), moderate (29-31 °C, 80%; THI = 81 to 83), and severe (32-34 °C, 80%; THI = 89 to 91) stress levels in climatic controlled chambers. Feed and water intake were recorded daily, and body weight was measured once a week. Blood was sampled every three days to analyse metabolite parameters. Dry matter intake (DMI) (p = 0.069, tendency) and blood glucose levels (p = 0.028) were decreased after sudden exposure to HS conditions (severe THI levels). Also, blood cortisol (p = 0.002), glutamic-oxaloacetic transaminase (GOT) (p = 0.009), blood urea nitrogen (BUN) (p = 0.004) and standing time (p = 0.009) were increased in moderate and severe THI levels compared with threshold after exposure to HS conditions. However, in the severe THI group, blood cortisol (p < 0.05), glucose (p < 0.05), GOT (p < 0.05) and BUN (p < 0.05) levels were returned to normal range after 6-13 days of continuous HS exposure. In conclusion, DMI, blood cortisol, GOT, glucose, BUN, and standing time were closely associated with long-term HS condition in beef calves. In addition, calves exposed to HS modulated their physiological responses that resulted in the regulation of the pertinent blood metabolites in the blood to maintain homeostasis during the long-term HS.

Effects of Dietary Supplementation of Acetate and L-Tryptophan Conjugated Bypass Amino Acid on Productivity of Pre- and Post-Partum Dairy Cows and Their Offspring.

In this study, we investigated the effect of dietary supplementation with acetate and L-tryptophan-conjugated bypass amino acid (ACT) during late pregnancy on the production performance of cows pre- and postpartum and their offspring. Eight multiparous Holstein cows (calving date ±15 d, 2nd parity; n = 4) were supplied with diets without ACT supplementation (Control) or with 15 g/day ACT supplementation (ACT). The results showed that ACT improved the feed intake (FI) in dry cows. No differences in blood hematological parameters were found between the two groups of prepartum cows. The serum glutamic-oxaloacetic transaminase activity increased and the triglyceride concentration decreased in the ACT-treated group compared to the control group. In the postpartum cows, milk compositions were not affected by ACT supplementation. Saturated fatty acid (SFA) content in the colostrum was significantly lower in the ACT-treated group than in the control group. Serum glucose (GLC) level was significantly higher in the ACT-treated group than in the control group. Monocyte and GLC levels were lower in calves of groups where their dams had received ACT. Overall, we found higher FI in the dry cows, lower colostrum SFA levels, and heavier calf birth weight (5.5 kg) when the dams were supplemented with ACT, suggesting a positive nutrient compensation by ACT supplementation to dry cows.

Effect of a Rumen-Protected Microencapsulated Supplement from Linseed Oil on the Growth Performance, Meat Quality, and Fatty Acid Composition in Korean Native Steers.

We evaluated the effects of a rumen-protected microencapsulated supplement from linseed oil (MO) on ruminal fluid, growth performance, meat quality, and fatty acid composition in Korean native steers. In an in vitro experiment, ruminal fluid was taken from two fistulated Holstein dairy cows. Different levels of MO (0%, 1%, 2%, 3%, and 4%) were added to the diet. In an in vivo experiment, eight steers (average body weight = 597.1 ± 50.26 kg; average age = 23.8 ± 0.12 months) were assigned to two dietary groups, no MO (control) and MO (3% MO supplementation on a DM basis), for 186 days. The in vitro study revealed that 3% MO is an optimal dose, as there were decreases in the neutral detergent fiber and acid detergent fiber digestibility at 48 h (p < 0.05). The in vivo study showed increases in the feed efficiency and average daily gain in the 3% MO group compared to the control group on days 1 to 90 (p < 0.05). Regarding meat quality, the shear force produced by the longissimus thoracis muscle in steers from the 3% MO group was lower than that produced by the control group (p < 0.05). Interestingly, in terms of the fatty acid profile, higher concentrations of C22:6n3 were demonstrated in the subcutaneous fat and higher concentrations of C18:3n3, C20:3n3, and C20:5n3 were found in the intramuscular fat from steers fed with 3% MO (p < 0.05). Our results indicate that supplementation with 3% MO supplements improves the growth performance and meat quality modulated by the omega-3 fatty acid content of meat in Korean native steers.

Effect of water scarcity during thermal-humidity exposure on the mineral footprint of sheep.

OBJECTIVE: Combination of two stressors on alteration of mineral footprints in animals needs due attention to meet maximum production and welfare, particularly in grazing sheep. This study tested whether ewes (Ovis aries) exposed to water deprivation and thermal-humidity stressors had altered mineral footprints in their wool, serum, urine, and feces. METHODS: Nine ewes (age = 3 years; mean body weight = 41±3.5 kg) were divided among a control group with free access to water, and treatment groups with water deprivation lasting either 2 h (2hWD) or 3 h (3hWD) after feeding. Using a 3×3 Latin square design, animals were assigned to treatment groups for three sampling periods of 21 days each (n = 9). Blood was collected by jugular venipuncture. Wool was collected at the end of periods 2 and 3. Metabolic crates designed with metal grated floors were used for urine and feces collection. We measured sodium (Na), magnesium (Mg), phosphorus (P), chloride (Cl), calcium (Ca), manganese (Mn), copper (Cu), iron (Fe), and zinc (Zn). RESULTS: The wool mineral levels did not differ between the treatment groups, although K was marginally lower (p = 0.10) in the 2hWD group. The serum and urine mineral levels did not differ between the treatments (p>0.05). Fecal K was significantly lower in the 2hWD group than in the other groups (p≤0.05). CONCLUSION: In conclusion, water deprivation and thermal-humidity exposure altered the excretion of K, but not of other minerals, in the wool, urine, feces, or serum of ewes. Thus, no additional mineral supplementation is needed for water deprived ewes during thermalhumidity exposure.

Heat-Shock Proteins Gene Expression in Peripheral Blood Mononuclear Cells as an Indicator of Heat Stress in Beef Calves.

This study was conducted to investigate the effect of HS on HSPs gene expression in bovine PBMCs of beef calves in in vitro and in vivo models. In the in vitro experiment, blood samples were collected from the jugular vein of five beef calves (age: 174.2 ± 5.20 days, BW: 145.2 ± 5.21 kg). In the in vivo experiment, sixteen Korean native male beef calves (age: 169.6 ± 4.60 days, BW: 136.9 ± 6.23 kg) were exposed to ambient temperature for seven days (22 to 24 °C, relative humidity 60%; temperature-humidity index (THI) = 68 to 70) and subsequently to the temperature and humidity corresponding to the target THI level for 21 days (HS). For PBMC isolation, blood samples were collected every three days. In the in vitro model, the cell viability was significantly decreased in HS groups compared with the control group (p = 0.015). The expression of HSP70 (p = 0.022), HSP90 (p = 0.003) and HSPB1 (p = 0.026) genes was increased in the HS group in in vitro model. In the in vivo experiment, the HSP70 gene expression was increased after sudden exposure to HS conditions (severe THI levels; THI = 88 to 90), whereas HSP90 and HSPB1 showed no differences among the THI groups (p > 0.05). However, in the severe THI group, the HSP70 gene expression returned to normal range after six days of continuous HS. In conclusion, the HSP70 gene plays a pivotal role in protecting cells from damage and is sensitive to HS in immune cells compared with other HSP genes in in vitro and in vivo models. In addition, the in vivo models suggest that calves exhibit active physiological mechanisms of adaptation to HS after six days of continuous exposure by regulating the HSP70 gene expression.

The effects of vitamin A supplementation during late-stage pregnancy on longissimus dorsi muscle tissue development, birth traits, and growth performance in postnatal Korean native calves.

OBJECTIVE: This study investigated the effects of vitamin A (VA) supplementation during late-stage pregnancy on longissimus dorsi muscle tissue development, birth traits, and growth performance of postnatal Korean native calves. METHODS: In the preliminary experiment, twenty-six pregnant cattle (initial body weight [BW] = 319 kg (standard deviation [SD] = 30.1; 1st parity) were randomly assigned to the control and treatment groups. The treatment group received VA supplementation at 24,000 IU/d from gestational day 225 until delivery. In the main experiment, twelve pregnant cattle (initial BW = 317 kg [SD = 31.3]; 1st parity) were treated with VA supplementation at 24,000 IU/d (gestational days 150 to 225) and at 78,000 IU/d (gestational day 225 until delivery). Serum VA levels were analyzed in pregnant cattle, and the growth performance, gene expression, and serum VA levels were analyzed in the offspring. RESULTS: Serum VA levels in pregnant cattle decreased the late gestation in both experiments (p<0.001). In the main experiment, pregnant cattle at parturition and offspring at birth in the treatment group had higher serum VA levels than those in the control group (p<0.05). In the treatment groups, an increased birth weight was observed in the main experimental group (p = 0.022), and a tendency (p = 0.088) toward an increased birth weight was observed in the preliminary experimental group. However, no differences were observed in the feed intake, average daily gain, gain-to-feed ratio, or BW of 31-day-old calves. Gene expression was analyzed in longissimus dorsi muscles of 31-day-old calves. VA supplementation in pregnant cattle stimulated postnatal muscle development in offspring by elevating myogenic factor 5 (MYF5), MYF6, and myoblast determination levels (p<0.05). Moreover, preadipocyte-related marker genes such as extracellular signal-regulated kinase 2 and krüppel-like factor 2 were higher in the treatment group than in the control group (p<0.05). CONCLUSION: VA supplementation (78,000 IU/d) in late-stage pregnant cattle maintained serum VA levels. In addition, 78,000 IU/d VA supplementation increased the birth weight and expression of genes related to muscle and preadipocyte development in offspring. Overall, 78,000 IU/d VA supplementation in pregnant cattle is beneficial to newborn calves.

Effect of Saccharomyces boulardii Supplementation on Performance and Physiological Traits of Holstein Calves under Heat Stress Conditions.

The objective of this study was to determine the effects of Saccharomyces boulardii CNCM I-1079 (SB) as a feed additive on performance, diarrhea frequency, rectal temperature, heart rate, water consumption, cortisol level, and fecal bacteria population in Holstein calves (28 ± 1.6 days of age, body weight of 45.6 ± 1.44 kg, n = 16) under thermal neutral (TN) and heat stress (HS) conditions. During the TN period for 21 days (d 1 to 21), calves receiving SB showed quadratic or linear effects compared to the control group, showing higher dry matter intake (DMI, p = 0.002), and water consumption (p = 0.007) but lower frequency of fecal diarrhea (p = 0.008), rectal temperature (p < 0.001), heart rate (p < 0.001), and fecal microbiota at 21 day (Escherichia coli, p = 0.025; Enterobacteriaceae, p = 0.041). Meanwhile, calves exposed to HS for 7 days (d 22 to 28) receiving SB showed quadratic or linear effects compared to the control group, showing higher DMI (p = 0.002) but lower water consumption (p = 0.023), rectal temperature (p = 0.026), and cortisol level (p = 0.014). Our results suggest that live SB is useful in the livestock industry as an alternative to conventional medication (especially in times of suspected health problems) that can be added to milk replacer for young dairy calves experiencing HS.