Dietary effects of protected fat, glycerol, and soybean meal on performance, physiological parameters, carcass characteristics, and behavioral measurements of late-fattening Hanwoo steers under heat stress conditions.

Objective: This study aimed to determine the effects of increasing energy and protein levels in diets by including protected fat (PF), glycerol (GL), and soybean meal (SBM) on growth performance, physiological parameters, carcass characteristics, and behavioral measurements of late-fattening Hanwoo steers under heat stress conditions. Methods: Thirty-six steers (initial BW, 724.9 ± 58.3 kg; age, 25.5 ± 0.4 month) were assigned into Control (TDN 76%, CP 15%), PF (TDN 83.6%, CP 15%), PF+GL (TDN 83.6%, CP 15%) and PF+GL+SBM (TDN 83.6%, CP 16.5%) by randomized complete block design for a total of 16 weeks with division of 4 week periods. The average temperature-humidity index was 87.0 (1st period; Severe), 82.8 (2nd; Moderate), 71.4 (3rd; Comfort), and 68.1 (4th; Comfort). Results: The DMI showed no treatments differences during the whole experiment. However, DMI in 1st and 2nd period decreased by approximately 30% and 10% compared to 4th period, respectively. Higher ADG and FCR were noted for treatments compared to Control at both 1st and 2nd period (p<0.05). There were no treatment effects on rectal temperature (RT), cortisol, and behaviors during the entire experiment. However, both RT and cortisol in 0, 1st and 2nd period were higher than those of 3rd and 4th period (p<0.05). Carcass yield and grade remained unaffected by increasing TDN and CP levels. Behavioral changes in the hot season (1st period) included reduced lying (43%), increased standing (48%), decreased walking (62%), and decreased eating (38%) (p<0.05), with an increase in drinking by 54%. Rumination during standing was 53% higher, while rumination during lying was about 33% lower compared to the post-hot season (3rd period) (p<0.05). Conclusion: Dietary supplementation of protected fat in late-fattening Hanwoo steers under heat stress had a positive effect on preventing a reduction in performance.

Inflammatory response in dairy cows caused by heat stress and biological mechanisms for maintaining homeostasis.

Climate change increases global temperatures, which is lethal to both livestock and humans. Heat stress is known as one of the various livestock stresses, and dairy cows react sensitively to high-temperature stress. We aimed to better understand the effects of heat stress on the health of dairy cows and observing biological changes. Individual cows were divided into normal (21-22 °C, 50-60% humidity) and high temperature (31-32 °C, 80-95% humidity), respectively, for 7-days. We performed metabolomic and transcriptome analyses of the blood and gut microbiomes of feces. In the high-temperature group, nine metabolites including linoleic acid and fructose were downregulated, and 154 upregulated and 72 downregulated DEGs (Differentially Expressed Genes) were identified, and eighteen microbes including Intestinimonas and Pseudoflavonifractor in genus level were significantly different from normal group. Linoleic acid and fructose have confirmed that associated with various stresses, and functional analysis of DEG and microorganisms showing significant differences confirmed that high-temperature stress is related to the inflammatory response, immune system, cellular energy mechanism, and microbial butyrate production. These biological changes were likely to withstand high-temperature stress. Immune and inflammatory responses are known to be induced by heat stress, which has been identified to maintain homeostasis through modulation at metabolome, transcriptome and microbiome levels. In these findings, heat stress condition can trigger alteration of immune system and cellular energy metabolism, which is shown as reduced metabolites, pathway enrichment and differential microbes. As results of this study did not include direct phenotypic data, we believe that additional validation is required in the future. In conclusion, high-temperature stress contributed to the reduction of metabolites, changes in gene expression patterns and composition of gut microbiota, which are thought to support dairy cows in withstanding high-temperature stress via modulating immune-related genes, and cellular energy metabolism to maintain homeostasis.

Advances in Methane Emission Estimation in Livestock: A Review of Data Collection Methods, Model Development and the Role of AI Technologies.

This review examines the significant role of methane emissions in the livestock industry, with a focus on cattle and their substantial impact on climate change. It highlights the importance of accurate measurement and management techniques for methane, a potent greenhouse gas accounting for 14-16% of global emissions. The study evaluates both conventional and AI-driven methods for detecting methane emissions from livestock, particularly emphasizing cattle contributions, and the need for region-specific formulas. Sections cover livestock methane emissions, the potential of AI technology, data collection issues, methane's significance in carbon credit schemes, and current research and innovation. The review emphasizes the critical role of accurate measurement and estimation methods for effective climate change mitigation and reducing methane emissions from livestock operations. Overall, it provides a comprehensive overview of methane emissions in the livestock industry by synthesizing existing research and literature, aiming to improve knowledge and methods for mitigating climate change. Livestock-generated methane, especially from cattle, is highlighted as a crucial factor in climate change, and the review underscores the importance of integrating precise measurement and estimation techniques for effective mitigation.

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.

Effects of heat stress on performance, physiological parameters, and blood profiles of early-fattening Hanwoo steers in climate chambers.

OBJECTIVE: This study was conducted to assess effects of heat stress on growth performance, physiological parameters, and blood profiles of Hanwoo steers during early-fattening period in climate chambers. METHODS: Four Hanwoo steers (body weight, 454.3±10.9 kg; age, 14±0.1 month) were allocated into four levels of temperature-humidity index (THI) in a 4×4 Latin square design for 21 days (pre-adaptation, 7 d; heat stress, 7 d; post-adaptation, 7 d) per period. Experimental treatments were assigned according to THI chart based on National Institute Animal Science (NIAS, 2022): Comfort (25.5°C to 26.5°C, 60%; THI 73 to 75), Mild (28°C to 29°C, 60%; THI 77 to 79), Moderate (29.5°C to 30.5°C, 80%; THI 82 to 84), and Severe (31°C to 32°C, 80%; THI 85 to 86) in separate climatic controlled chambers. RESULTS: The dry matter intake (DMI) of the formula feed was lower in Severe compared to Mild and Comfort (p<0.05). The DMI of rice straw was the lowest in Severe and lower in Moderate than Comfort and Mild (p<0.05). Both average daily gain and feed conversion ratio of Severe and Moderate were lower than those of Mild and Comfort (p<0.05). Water intake was the highest in Severe and lower in Moderate compared with Comfort and Mild (p<0.05). Heart rate and rectal temperature increased as THI level increased (p<0.05). Glucose was the lowest in Severe and lower in Moderate compared to Comfort (p<0.05). On the contrary, non-esterified fatty acid was the highest in Severe and lower in Moderate compared with Comfort (p<0.05). Blood urea nitrogen of Moderate and Severe were higher than those of Comfort and Mild (p<0.05). Cortisol increased as THI increased (p<0.05). CONCLUSION: This study demonstrated the negative effects of heat stress on the performance and physiological responses of Hanwoo steers during the early-fattening period. In addition, it is judged that the THI chart for Hanwoo steers of National Institute of Animal Science (2022) was properly calculated.

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.

Measuring the effects of estrus on rumen temperature and environment, behavior and physiological attributes in Korean Native breeding cattle.

In this study, rumen temperature and environment in estral and non-estral Korean Native breeding cattle were evaluated by using a bolus sensor. Behavioral and physiological changes in study animals were also assessed. To assess the rumen temperature and environment, we inserted bolus sensors into 12 Korean Native cattle with an average age of 35.5 months, then measured temperature and activity within the rumen using the wireless bolus sensor. Drinking, feeding and mounting behavior, and measured vaginal temperature and levels of intravaginal mucus resistance were recorded. We found that cattle in estrus exhibited more acts of mounting (37.4 vs. 0 times/day), increased vaginal temperature (39.0°C vs. 38.4°C), and decreased vaginal mucus resistance (136.3 Ω vs 197.4 Ω), compared with non-estral animals. Furthermore, increased levels of rumen activity were most significant in estrus cattle at the highest activity levels (p < 0.01). Overall, the estrus group exhibited increased rumen temperature (p = 0.01), compared with the non-estrus group. In conclusion, the results of this study not only provide basic physiological data related to estrus in improved Korean Native breeding cattle, but also suggest that monitoring of rumen temperature and activity might be used as an effective smart device for estrus detection.

Vitamin D3 decreases myoblast fusion during the growth and increases myogenic gene expression during the differentiation phase in muscle satellite cells from Korean native beef cattle.

The process of myogenesis, which involves the growth and differentiation of muscle cells, is a crucial determinant of meat yield and quality in beef cattle. Essential nutrients, such as vitamins D and A, play vital roles in the development and maintenance of various tissues, including muscle. However, limited knowledge exists regarding the specific effects of vitamins A and D in bovine muscle. Therefore, the aim of this study was to investigate the impact of vitamins A and D treatment on myogenic fusion and differentiation in bovine satellite cells (BSC). BSC were isolated from Korean native beef cattle, specifically from four female cows approximately 30 mo old. These individual cows were used as biological replicates (n = 3 or 4), and we examined the effects of varying concentrations of vitamins A (All-trans retinoic acid; 100 nM) and D (1,25-dihydroxy-vitamin D3; 1 nM, 10 nM, and 100 nM), both individually and in combination, on myoblast fusion and myogenic differentiation during the growth phase (48 h) or differentiation phase (6 d). The results were statistically analyzed using GLM procedure of SAS with Tukey's test and t-tests or one-way ANOVA where appropriate. The findings revealed that vitamin A enhanced the myoblast fusion index, while vitamin D treatment decreased the myoblast fusion index during the growth phase. Furthermore, vitamin A treatment during the differentiation phase promoted terminal differentiation by regulating the expression of myogenic regulatory factors (Myf5, MyoD, MyoG, and Myf6) and inducing myotube hypertrophy compared to the control satellite cells (P < 0.01). In contrast, vitamin D treatment during the differentiation phase enhanced myogenic differentiation by increasing the mRNA expression of MyoG and Myf6 (P < 0.01). Moreover, the combined treatment of vitamins A and D during the growth phase increased myoblast fusion and further promoted myogenic differentiation and hypertrophy of myotubes during the differentiation phase (P < 0.01). These results suggest that vitamin A and D supplementation may have differential effects on muscle development in Korean native beef cattle during the feeding process.

The study investigated the effects of vitamins A and D on the growth and differentiation phases of bovine satellite cells and found that both vitamins have a positive impact on muscle development. Vitamin A promoted myoblast fusion during the growth phase, leading to increased myotube formation, while vitamin D suppressed myoblast fusion during this phase. However, during the differentiation phase, both vitamins enhanced terminal differentiation and hypertrophy. Vitamin A promoted the activation of satellite cells, while vitamin D promoted the expression of genes that enhance myogenesis. The combination treatment of vitamins A and D during the growth phase complemented each other to increase myogenic cell fusion, and during differentiation, promoted terminal differentiation and hypertrophy. These findings suggest that supplementing cattle feed with both vitamins A and D has the potential to enhance muscle development, which would be advantageous for the meat industry.

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.

Iris Pallida Extract Alleviates Cortisol-Induced Decrease in Type 1 Collagen and Hyaluronic Acid Syntheses in Human Skin Cells.

Excessive endogenous or exogenous levels of the stress hormone cortisol have negative effects on various tissues, including the skin. Iris pallida (IP), used in traditional medicine and perfumes, exhibits biological activities, such as antioxidant and anti-inflammatory activities. In this study, we aimed to investigate the inhibitory effect of IP extract (IPE) on cortisol activity in human skin cells. We found that IPE alleviated the cortisol-induced decrease in the levels of procollagen type 1 and hyaluronic acid (HA), which were significantly recovered by 106% and 31%, respectively, compared with cortisol-induced reductions. IPE also rescued the suppression of the gene expression of COL1A1 and the HA synthases HAS2 and HAS3 in cortisol-exposed cells. Moreover, IPE blocked the cortisol-induced translocation of the glucocorticoid receptor (GR) from the cytoplasm to the nucleus as effectively as the GR inhibitor mifepristone. Analysis using a high-performance liquid chromatography-diode-array detector system revealed that irigenin, an isoflavone, is the main component of IPE, which restored the cortisol-induced reduction in collagen type 1 levels by 82% relative to the cortisol-induced decrease. Our results suggest that IPE can act as an inhibitor of cortisol in human skin cells, preventing cortisol-induced collagen and HA degradation by blocking the nuclear translocation of the GR. Therefore, IPE may be used as a cosmetic material or herbal medicine to treat stress-related skin changes.

Supplementation of complex natural feed additive containing (C. militaris, probiotics and red ginseng by-product) on rumen-fermentation, growth performance and carcass characteristics in Korean native steers.

This study evaluated the effects of a complex natural feed additive on rumen fermentation, carcass characteristics and growth performance in Korean-native steers. In this study, in vitro and in vivo experiment were conducted. Seven different levels of complex natural feed additive (CA) were added to the buffered rumen fluid using AnkomRF gas production system for 12, 24 and 48 h. All experimental data were analyzed by mixed procedure of SAS. Total gas production increased in the CA groups, with the highest response observed in the 0.06% group at 48 h of incubation (linear, p = 0.02; quadratic, p < 0.01). Regarding rumen fermentation parameters, the total volatile fatty acid (TVFA) tended to increase in all the CA groups (p = 0.07). The concentrations of butyrate, iso-butyrate, and iso-valerate significantly increased in all treatment groups (p < 0.05). In the in vivo experiment, 23 Korean-native steers were allocated to two groups: (1) Control and (2) Treatment; control +0.07% CA (DM basis), in a randomized complete-block design and blocked by body weight (ave. body weight = 641.96 kg ± 62.51 kg, p = 0.80) and feed intake (ave. feed intake = 13.96 kg ± 0.74 kg, p = 0.08) lasted for 252 days. Average daily gain decreased in the treatment group (p < 0.01). Backfat thickness significantly decreased in the CA group (p = 0.03), whereas meat color tended to increase (p = 0.07). In conclusion, in the in vitro experiment, the inclusion of complex natural feed additive decreased methane proportion and tended to increase TVFA production, but supplementation to Korean native steers decreased average daily gain and backfat thickness.

Stress Concepts and Applications in Various Matrices with a Focus on Hair Cortisol and Analytical Methods.

When studying stress in animals, it is important to understand the types of stress and their classification, and how to assess the stress levels in different animal species using different matrices accurately and precisely. The classification of stress types helps to distinguish between good stress (eustress) and bad stress (distress). Hence, first, it is crucial to assess the animal's level of stress in a non-intrusive manner and second to identify the type of stress that is best suited to its environment. Third, it is also important to analyze the obtained samples using a suitable method to increase the validity of stress hormone measurements. Therefore, in this review, we aim to: (1) explain the classification of stress, (2) discuss the wide range of body matrices (e.g., saliva, milk, hair, urine, feces, sweat, fins, etc.) that can be used as samples to evaluate stress levels, as well as their comparisons and limitations, and present the reliable matrices for measuring stress hormones with special emphasis on hair, (3) compare the analytical methods for measuring stress hormones after sample preparation. Despite some literature that does not include hair as a reliable matrix for evaluating stress levels, hair is one of the matrices for measuring long-term stress hormone accumulations. This review discusses some factors that influence the level of stress hormones in the hair. By understanding these issues, the scientific community will not only be able to improve the understanding of stress and biomarker evaluation but also suggest how to deal with the consequences of stress in future research.

Efficient Myogenic/Adipogenic Transdifferentiation of Bovine Fibroblasts in a 3D Bioprinting System for Steak-Type Cultured Meat Production.

The interest in cultured meat is increasing because of the problems with conventional livestock industry. Recently, many studies related to cultured meat have been conducted, but producing large-sized cultured meat remains a challenge. It is aimed to introduce 3D bioprinting for producing large cell aggregates for cultured meat production. A hydrogel scaffold is produced at the centimeter scale using a bioink consisting of photocrosslinkable materials for digital light processing-based (DLP) printing, which has high printing accuracy and can produce geometrically complex structures. The light exposure time for hydrogel photopolymerization by DLP bioprinting is optimized based on photorheometry and cell viability assays. Naturally immortalized bovine embryonic fibroblast cells transformed with MyoD and PPARγ2 instead of primary cells are used as the latter have difficulties in maintaining stemness and are associated with animal ethics issues. The cells are mixed into the hydrogel for printing. Myogenesis and adipogenesis are induced simply by changing the medium after printing. Scaffolds are obtained successfully with living cells and large microchannels. The cooked cultured meat maintains its size and shape upon cutting. The overall dimensions are 3.43 cm × 5.53 cm × 0.96 cm. This study provides proof-of-concept for producing 3D cultured meat using bioinks.

Evaluation of Heat Stress Effects in Different Geographical Areas on Milk and Rumen Characteristics in Holstein Dairy Cows Using Robot Milking and Rumen Sensors: A Survey in South Korea.

This survey investigated, using robotic milking and rumen sensors, the effects of an adjusted temperature−humidity index (THI) in different geographical areas on milk yield, fat and protein, rumen temperature, and activity in lactating Holstein cows. We additionally explored the effect of parity on milk and rumen temperature and activity under different THI levels during the summer. From January to September 2020, four farms (276 dairy cows) were subjected to the use of robot milking machines, and two farms (162 dairy cows) to the use of rumen sensors. For the temperature and humidity data, the THI was calculated on the basis of the data from the Korea Meteorological Administration (KMA). The data were analyzed using the GLM procedure of SAS. Milk yield and milk protein decreased (p < 0.05), and milk fat increased (p < 0.05) at all farms during the summer, from July to August, when the temperature and humidity were high (THI = 72−79). Milk yields were the highest in the fifth, sixth, seventh, and eighth parities, and the lowest in the fourth (p < 0.05). Milk fat concentration was the highest in the fourth parity and the lowest in the first parity (p < 0.05). In the first parity, the highest levels of milk protein and lactose were seen (5.24% and 4.90%, respectively). However, milk protein concentration was the lowest in the third parity, and the lactose concentration was the lowest in the fifth, sixth, seventh, and eighth parities. According to the rumen sensor, the rumen temperature of the dairy cows at the two farms also continued to increase (p < 0.05) from July to August, and then decreased (p < 0.05) in September. However, the activity in the rumen was increased (p < 0.05) from July to September. In the second parity, the highest rumen temperature (39.02 °C) was observed, while the lowest value (38.28 °C) was observed in the third parity. The highest value of rumen activity (12.26 mg) was observed in the second parity and the lowest value (11.31 mg) in the fourth parity. These data, taken together, confirm that a high THI during summer conditions negatively affects milk yield, milk protein content, and rumen temperature and activity in lactating Holstein cows. It is also demonstrated that various parities affect milk characteristics and the rumen environment in the summer season.

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.

Vitamin A supplementation downregulates ADH1C and ALDH1A1 mRNA expression in weaned beef calves.

Our previous studies demonstrated that oral vitamin A supplementation during late-stage pregnancy and the neonatal stage enhances birth weight, growth performance, and mRNA expression related to muscle and preadipocyte development in beef cattle. The alcohol dehydrogenase 1C (ADH1C) c.-64T > C genotype also correlated with vitamin A concentration in beef production. This study aimed to investigate the effects of vitamin A supplementation on the muscle development and vitamin A metabolism in weaned beef calves with different ADH1C genotypes. Twenty male calves (90 d of age; initial BW: 89.03 kg [SD 8.60]) were stratified according to ADH1C genotype and vitamin A treatment (duration: 3 months) and randomly assigned to 4 groups with a 2 × 2 factorial arrangement. Vitamin A treatments included the following: control (10,000 IU/kg of as-fed, a. TT type; b. TC type); treatment (40,000 IU/kg of as-fed, c. TT type; and d. TC type). Parameters including BW, FI, blood, longissimus dorsi muscle, and liver status during the experimental period were analyzed using the generalized linear model (GLM) procedure and Tukey's test by SAS 9.4 program. Serum vitamin A was significantly increased (P < 0.05) in the vitamin A treatment group at 4 and 6 months of age. TT type calves showed higher serum vitamin A concentration (P < 0.05) than the TC type calves. Serum triglyceride and non-esterified fatty acid (NEFA) levels increased (P < 0.05) in the treatment group compared with the control at 6 months of age. However, BW, ADG and FI showed no differences between the groups. In addition, mRNA expression in longissimus dorsi muscle revealed upregulation of paired box 7 (PAX7) (P < 0.05) after the vitamin A treatment period based on biopsy results. Both ADH1C and aldehyde dehydrogenase (ALDH) 1A1 mRNA expression was downregulated (P < 0.01) by vitamin A supplementation. The TC type of ADH1C showed higher mRNA expression than the TT type. However, no effect was observed on adipogenic mRNA expression (preadipocyte factor-1 [PREF-1], peroxisome proliferator-activated receptor gamma [PPARγ], fatty acid binding protein 4 [FABP4]) in all groups. Our findings suggest that weaned calves treated with vitamin A may promote the storage of satellite cells by elevating PAX7 gene expression in the muscle. The TC type calves may show increased capacity for vitamin A metabolism, which can be used in genetically customizing feed management to maximize beef production in the calves.

D-Methionine and 2-hydroxy-4-methylthiobutanoic acid i alter beta-casein, proteins and metabolites linked in milk protein synthesis in bovine mammary epithelial cells.

This study aims to determine the effects of D-methionine (D-Met) isomer and the methionine precursor 2-hydroxy-4-methylthiobutanoic acid i (HMBi) supplementation on milk protein synthesis on immortalized bovine mammary epithelial cell (MAC-T). MAC-T cells were seeded using 10-cm dishes and cultured in Dulbecco's modified Eagle's medium/F12 (DMEM/F12) basic medium. The basic medium of DMEM/F12 was replaced with the lactogenic DMEM/F12 differentiation medium when 90% of MAC-T cells reached confluency. The best dosage at 0.6 mM of D-Met and HMBi and incubation time at 72 h were used uniformly for all treatments. Each treatment was replicated six times wherein treatments were randomly assigned in a 6-well plate. Cell, medium, and total protein were determined using a bicinchoninic acid protein assay kit. Genes, proteomics and metabolomics analyses were also done to determine the mechanism of the milk protein synthesis pathway. Data were analyzed by two-way analysis of variance (ANOVA) with supplement type and plate as fixed effects. The least significant difference test was used to evaluate the differences among treatments. The HMBi treatment group had the highest beta-casein and S6 kinase beta-1 (S6K1) mRNA gene expression levels. HMBi and D-Met treatments have higher gene expressions compared to the control group. In terms of medium protein content, HMBi had a higher medium protein quantity than the control although not significantly different from the D-Met group. HMBi supplementation stimulated the production of eukaryotic translation initiation factor 3 subunit protein essential for protein translation initiation resulting in higher medium protein synthesis in the HMBi group than in the control group. The protein pathway analysis results showed that the D-Met group stimulated fructose-galactose metabolism, glycolysis pathway, phosphoinositide 3 kinase, and pyruvate metabolism. The HMBi group stimulated the pentose phosphate and glycolysis pathways. Metabolite analysis revealed that the D-Met treatment group increased seven metabolites and decreased uridine monophosphate (UMP) production. HMBi supplementation increased the production of three metabolites and decreased UMP and N-acetyl-L-glutamate production. Taken together, D-Met and HMBi supplementation are effective in stimulating milk protein synthesis in MAC-T cells by genes, proteins, and metabolites stimulation linked to milk protein synthesis.