Dietary supplementation of phytoncide and soybean oil increases milk conjugated linoleic acid and depresses methane emissions in Holstein dairy cows.

The objective of this study was to determine whether adding phytoncide oil (PO) and soybean oil (SBO) to the dairy cow diet could increase milk conjugated linoleic acid (CLA) and depress methane (CH4) emissions in Holstein dairy cows. Rumen fermentation was conducted at four levels of SBO (0, 1, 2, and 4%, on DM basis) and two levels of PO (0 and 0.1%, on DM basis) with in vitro experiment. To evaluate blood parameters, fecal microbe population, milk yield and fatty acid compositions, and CH4 production, in vivo experiment was conducted using 38 Holstein dairy cows divided into two groups of control (fed TMR) and treatment (fed TMR with 0.1% PO and 2% SBO as DM basis). In the in vitro study (Experiment 1), PO or SBO did not affect rumen pH. However, SBO tended to decrease ruminal ammonia-N (p = 0.099). Additionally, PO or SBO significantly decreased total gas production (p = 0.041 and p = 0.034, respectively). Both PO and SBO significantly decreased CH4 production (p < 0.05). In addition, PO significantly increased both CLA isomers (c9, t11 and t10, c12 CLA) (p < 0.001). Collectively, 0.1% PO and 2% SBO were selected resulting in most effectively improved CLA and decreased CH4 production. In the in vivo study (Experiment 2), 0.1% PO with 2% SBO (PSO) did not affect complete blood count. However, it decreased blood urea nitrogen and magnesium levels in blood (p = 0.021 and p = 0.01, respectively). PSO treatment decreased pathogenic microbes (p < 0.05). It increased milk yield (p = 0.017) but decreased percentage of milk fat (p = 0.013) and MUN level (p < 0.01). In addition, PSO treatment increased both the concentration of CLA and PUFA in milk fat (p < 0.01). Finally, it decreased CH4 emissions from dairy cows. These results provide compelling evidence that a diet supplemented with PSO can simultaneously increase CLA concentration and decrease CH4 production with no influence on the amount of milk fat (kg/day) in Holstein dairy cows.

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.

Quantitative Associations between Season, Month, and Temperature-Humidity Index with Milk Yield, Composition, Somatic Cell Counts, and Microbial Load: A Comprehensive Study across Ten Dairy Farms over an Annual Cycle.

This current study addresses the knowledge gap regarding the influence of seasons, months, and THI on milk yield, composition, somatic cell counts (SCC), and total bacterial counts (TBC) of dairy farms in northeastern regions of Iran. For this purpose, ten dairy herds were randomly chosen, and daily milk production records were obtained. Milk samples were systematically collected from individual herds upon delivery to the dairy processing facility for subsequent analysis, including fat, protein, solids-not-fat (SNF), pH, SCC, and TBC. The effects of seasons, months, and THI on milk yield, composition, SCC, and TBC were assessed using an analysis of variance. To account for these effects, a mixed-effects model was utilized with a restricted maximum likelihood approach, treating month and THI as fixed factors. Our investigation revealed noteworthy correlations between key milk parameters and seasonal, monthly, and THI variations. Winter showed the highest milk yield, fat, protein, SNF, and pH (p < 0.01), whereas both SCC and TBC reached their lowest values in winter (p < 0.01). The highest values for milk yield, fat, and pH were recorded in January (p < 0.01), while the highest protein and SNF levels were observed in March (p < 0.01). December marked the lowest SCC and TBC values (p < 0.01). Across the THI spectrum, spanning from -3.6 to 37.7, distinct trends were evident. Quadratic regression models accounted for 34.59%, 21.33%, 4.78%, 20.22%, 1.34%, 15.42%, and 13.16% of the variance in milk yield, fat, protein, SNF, pH, SCC, and TBC, respectively. In conclusion, our findings underscore the significant impact of THI on milk production, composition, SCC, and TBC, offering valuable insights for dairy management strategies. In the face of persistent challenges posed by climate change, these results provide crucial guidance for enhancing production efficiency and upholding milk quality standards.

Worldwide Research Trends for Chelates in Animal Science: A Bibliometric Analysis.

The purpose of this study was to look at research trends in the application of CTM in animal nutrition in order to identify current and emerging challenges, as well as to examine the intellectual structure of the subject. The intellectual structure of CTM was examined using keyword and reference analysis. The research community includes all research and review articles published in journals indexed in the Web of Science database during the years 1990-2022. The results showed that the terms zinc, co-occurring 331 times, performance (324 times), and copper 216 (times) were the main and hotspots of research in the field of chelate. The data suggest that the most important keywords during the study period were zinc, copper, pig, bovine, metabolism, and bioavailability. The terms health, muscle, beef, trace elements, and dietary supplements represent emerging topics in CTM, as research began to focus on these areas during the years 2017-2022. The country with the greatest number of published articles was the United States of America. This bibliometric analysis showed that countries are focusing on the effects of CTM on the health and musculature of cattle through dietary supplementation with trace elements. According to the identified hot and emerging topics, this research can serve as a roadmap for a global comprehensive scientific plan and policy.

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.

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.

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 Environmental Temperature and Humidity on Milk Composition, Microbial Load, and Somatic Cells in Milk of Holstein Dairy Cows in the Northeast Regions of Iran.

The present study aims to examine the relationships between temperature and humidity and milk composition, microbial load, and somatic cells in the milk of Holstein dairy cows. For this purpose, the temperature−humidity index, ambient temperature, and relative humidity data were obtained from the nearest weather stations. Production data were obtained from four dairy farms in Golestan province, Iran, collected from 2016 to 2021. The traits investigated were protein, fat, solids-not-fat (SNF), microbial load, and somatic cell count (SCC) in milk. The effects of the environmental temperature, humidity, month, and season on the milk composition, microbial load, and somatic cells were analyzed through analysis of variance. The effects of environmental temperature, humidity, month, and season on the milk composition, microbial load, and somatic cell composition were analyzed using a mixed procedure with a restricted maximum likelihood model. Although our findings revealed that there were significant differences in fat, protein, SNF, and SCC among the different months of the year (p < 0.01), no significant difference was observed in the total microbial count in milk. Environmental temperature presented significant impacts on fat, protein, SNF, SCC, and total microbial count within various temperature ranges (p < 0.01). When the temperature increased from 6.2 °C to 31.3 °C, the milk protein, fat, SNF, and somatic cell count significantly decreased, by approximately 4.09%, 5.75%, 1.31%, and 16.8%, respectively; meanwhile, the microbial count in milk significantly increased, by approximately 13.7%. Humidity showed an influence on fat, protein, non-fat solids, somatic cells, and total microbial count within different temperature ranges (p < 0.01). When the humidity increased from 54% to 82%, the milk protein, fat, SNF, and SCC significantly increased, by approximately 3.61%, 4.84%, 1.06%, and 10.2%, respectively; meanwhile, the microbial count in milk significantly decreased, by approximately 16.3%. The results demonstrate that there is a negative correlation between different months of the year, temperature, and the humidity of the environment, in terms of milk components and SCC. Our findings demonstrate that the optimum performance, in terms of milk composition, occurred in the first quarter of the year. As temperature increases and humidity decreases, milk quality decreases. Therefore, the adverse effects of environmental conditions on agricultural profits are not negligible, and strategies to better deal with the negative environmental effects are needed in order to improve milk quality in dairy cows.

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.

Milk Supplemented with Organic Iron Improves Performance, Blood Hematology, Iron Metabolism Parameters, Biochemical and Immunological Parameters in Suckling Dalagh Lambs.

This study was conducted to investigate the effect of milk supplemented with organic iron on performance, blood hematology, iron metabolism parameters, biochemical and immunological parameters in suckling lambs. Thirty-six newborn Dalagh lambs were randomly divided into three groups with 12 replications. The control group was fed with milk without organic iron. The other two groups were fed milk supplemented with 25 and 50 mg/d organic iron, respectively. During the experiment, increased daily weight gain and total body weight were observed in the iron-supplemented groups. An increase in the levels of red blood cell, hemoglobin, hematocrit, mean corpuscular hemoglobin, and mean corpuscular concentration in iron supplemented groups was indicated. Consumption of organic iron caused a significant decrease in plasma copper concentration. Total antioxidant status level was lower, but levels of glutathione peroxidase, superoxide dismutase, and catalase were higher in iron supplemented groups. In organic iron supplemented groups, insulin and thyroid hormones levels were significantly increased, and glucose level was significantly decreased. In organic iron supplemented groups, alkaline phosphatase level significantly increased, and aminotransferase level significantly decreased. Overall, the use of organic iron in the milk improved the performance and health in suckling lambs, and since a lower level of supplementation is naturally preferable, supplementation of milk with 25 mg/d organic iron is recommended.

Daytime Grazing in Mountainous Areas Increases Unsaturated Fatty Acids and Decreases Cortisol in the Milk of Holstein Dairy Cows.

The effects of grazing lactating cows in mountainous areas for 12 and 24 h compared with the confined indoor system were evaluated by examining the overall milk fatty acid and cortisol. Twenty-one dairy cows were allocated to three treatment groups: (1) control (confined management system in a free-stall barn; TMR based), (2) grazing for 12 h (12 hG; TMR plus grazing pasture), and (3) grazing for 24 h (24 hG; pasture-based feeding system). Dry matter intake was higher in the control and 12 hG groups than in the 24 hG group. The yields of total milk and the 3.5% fat-corrected milk were the lowest in the 24 hG group. Milk fat was the highest in the 24 hG group and higher in 12 hG compared with the control group. Milk protein and lactose levels were the highest in the 12 hG group. The highest somatic cell count was observed in the 24 hG group. The saturated fatty acid levels were higher in the control group compared with the 12 hG and 24 hG groups. There was no difference in overall mono-unsaturated fatty acids between 12 hG and 24 hG groups. Poly-unsaturated fatty acids were higher in the 12 hG group compared with the control and 24 hG groups. There was no difference in omega-6 (ω-6) fatty acids among the groups, and omega-3 fatty acids were higher in the 12 hG group than in the control group. Milk cortisol was the highest in the 24 hG group and higher in the control group compared with the 12 hG group. Taken together, grazing for 12 h is advisable for farms that have access to mountainous areas to improve the milk fatty acid profile and decrease the stress levels in high-yielding Holstein lactating cows.

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.

Dietary supplementation of acetate-conjugated tryptophan alters feed intake, milk yield and composition, blood profile, physiological variables, and heat shock protein gene expression in heat-stressed dairy cows.

The purpose of this study was to investigate the effects of dietary supplementation of rumen-protected tryptophan (RPT) at four levels on milk yield, milk composition, blood profile, physiological variables, and heat shock protein gene expression in dairy cows under conditions of moderate-severe heat stress (MSHS, THI = 80~89). Sixteen early-lactating dairy cows (body weight = 719 ± 66.4 kg, days in milk = 74.3 ± 7.1, milk yield = 33.55 ± 3.74 kg, means ± SEM) were randomly assigned in a factorial arrangement to one of the four treatments: control group (n = 4, no RPT supplementation), 15 g/d RPT (n = 4), 30 g/d RPT (n = 4), or 60 g/d RPT group per cow (n = 4) supplemented to the TMR. A higher dry matter intake (DMI) and milk yield were found in the 30 g RPT group compared with the other groups, and the 3.5% fat-corrected milk yield, energy-corrected milk yield, milk fat, protein, ß-casein, mono-unsaturated fatty acid, and poly-unsaturated fatty acid contents, and serum glucose content were observed in the 30 g RPT group (p < 0.05). The milk lactose concentration was significantly higher in the 30 g RPT group compared with the control and 60 g RPT groups (p < 0.05). The plasma cortisol level was lower, while the serotonin and melatonin concentrations were higher in the 30 g group compared with the other groups (p < 0.05). Heat shock protein (HSP) 70 expression was downregulated in the control and 15 g RPT groups, whereas the expression of HSP90 and HSPB1 remained unchanged among the groups. In particular, the 30 g RPT group was considered to have an improved DMI, milk yield, and lactose concentration, as well as anti-heat stress effects due to the simulation of serotonin and melatonin during MSHS.

Supplementing with L-Tryptophan Increases Medium Protein and Alters Expression of Genes and Proteins Involved in Milk Protein Synthesis and Energy Metabolism in Bovine Mammary Cells.

The objective of this study was to investigate the effects of supplementing with L-tryptophan (L-Trp) on milk protein synthesis using an immortalized bovine mammary epithelial (MAC-T) cell line. Cells were treated with 0, 0.3, 0.6, 0.9, 1.2, and 1.5 mM of supplemental L-Trp, and the most efficient time for protein synthesis was determined by measuring cell, medium, and total protein at 0, 24, 48, 72, and 96 h. Time and dose tests showed that the 48 h incubation time and a 0.9 mM dose of L-Trp were the optimal values. The mechanism of milk protein synthesis was elucidated through proteomic analysis to identify the metabolic pathway involved. When L-Trp was supplemented, extracellular protein (medium protein) reached its peak at 48 h, whereas intracellular cell protein reached its peak at 96 h with all L-Trp doses. ß-casein mRNA gene expression and genes related to milk protein synthesis, such as mammalian target of rapamycin (mTOR) and ribosomal protein 6 (RPS6) genes, were also stimulated (p < 0.05). Overall, there were 51 upregulated and 59 downregulated proteins, many of which are involved in protein synthesis. The results of protein pathway analysis showed that L-Trp stimulated glycolysis, the pentose phosphate pathway, and ATP synthesis, which are pathways involved in energy metabolism. Together, these results demonstrate that L-Trp supplementation, particularly at 0.9 mM, is an effective stimulus in ß-casein synthesis by stimulating genes, proteins, and pathways related to protein and energy metabolism.

Characterization of Short-Term Heat Stress in Holstein Dairy Cows Using Altered Indicators of Metabolomics, Blood Parameters, Milk MicroRNA-216 and Characteristics.

This study aims to characterize the influence of short-term heat stress (HS; 4 day) in early lactating Holstein dairy cows, in terms of triggering blood metabolomics and parameters, milk yield and composition, and milk microRNA expression. Eight cows (milk yield = 30 ± 1.5 kg/day, parity = 1.09 ± 0.05) were homogeneously housed in environmentally controlled chambers, assigned into two groups with respect to the temperature humidity index (THI) at two distinct levels: approximately ~71 (low-temperature, low-humidity; LTLH) and ~86 (high-temperature, high-humidity; HTHH). Average feed intake (FI) dropped about 10 kg in the HTHH group, compared with the LTLH group (p = 0.001), whereas water intake was only numerically higher (p = 0.183) in the HTHH group than in the LTLH group. Physiological parameters, including rectal temperature (p = 0.001) and heart rate (p = 0.038), were significantly higher in the HTHH group than in the LTLH group. Plasma cortisol and haptoglobin were higher (p < 0.05) in the HTHH group, compared to the LTLH group. Milk yield, milk fat yield, 3.5% fat-corrected milk (FCM), and energy-corrected milk (ECM) were lower (p < 0.05) in the HTHH group than in the LTLH group. Higher relative expression of milk miRNA-216 was observed in the HTHH group (p < 0.05). Valine, isoleucine, methionine, phenylalanine, tyrosine, tryptophan, lactic acid, 3-phenylpropionic acid, 1,5-anhydro-D-sorbitol, myo-inositol, and urea were decreased (p < 0.05). These results suggest that early lactating cows are more vulnerable to short-term (4 day) high THI levels-that is, HTHH conditions-compared with LTLH, considering the enormous negative effects observed in measured blood metabolomics and parameters, milk yield and compositions, and milk miRNA-216 expression.

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 Dietary Rumen-Protected L-Tryptophan Supplementation on Growth Performance, Blood Hematological and Biochemical Profiles, and Gene Expression in Korean Native Steers under Cold Environment.

We assessed the growth performance, physiological traits, and gene expressions in steers fed with dietary rumen-protected L-tryptophan (RPT) under a cold environment. Eight Korean native steers were assigned to two dietary groups, no RPT (Control) and RPT (0.1% RPT supplementation on a dry matter basis) for six weeks. Maximum and minimum temperatures throughout the experiment were 6.7 °C and -7.0 °C, respectively. Supplementation of 0.1% RPT to a total mixed ration did not increase body weight but had positive effects of elevating average daily gain (ADG) and reducing the feed conversion ratio (FCR) on days 27 and 48. The metabolic parameter showed a higher glucose level (on day 27) in the 0.1% RPT group compared to the control group. Real-time PCR analysis showed no significant differences in the expression of muscle (MYF6, MyoD, and Desmin) metabolism genes between the two groups, whereas the expression of fat (PPARγ, C/EBPα, and FABP4) metabolism genes was lower in the 0.1% RPT group than in the control group. Thus, we demonstrate that long-term (six weeks) dietary supplementation of 0.1% RPT was beneficial owing to enhanced growth performance by increasing the ADG and glucose level, decreasing FCR, and maintaining homeostasis in immune responses in beef steers in a cold environment.

Cortisol Extraction from Sturgeon Fin and Jawbone Matrices.

The aims of this study were to develop a technique for the extraction of cortisol from sturgeon fins using two washing solvents (water and isopropanol) and quantify any differences in fin cortisol levels among three main sturgeon species. Fins were harvested from 19 sacrificed sturgeons including seven beluga (Huso huso), seven Siberian (Acipenser baerii), and five sevruga (A. stellatus). The sturgeons were raised in Iranian farms for 2 years (2017-2018), and cortisol extraction analysis was conducted in South Korea (January-February 2019). Jawbones from five H. huso were also used for cortisol extraction. Data were analyzed using the general linear model (GLM) procedure in the SAS environment. The intra- and inter-assay coefficients of variation were 14.15 and 7.70, respectively. Briefly, the cortisol extraction technique involved washing the samples (300 ± 10 mg) with 3 mL of solvent (ultrapure water and isopropanol) twice, rotation at 80 rpm for 2.5 min, air-drying the washed samples at room temperature (22-28 °C) for 7 days, further drying the samples using a bead beater at 50 Hz for 32 min and grinding them into powder, applying 1.5 mL methanol to the dried powder (75 ± 5 mg), and slow rotation (40 rpm) for 18 h at room temperature with continuous mixing. Following extraction, samples were centrifuged (9,500 x g for 10 min), and 1 mL supernatant was transferred into a new microcentrifuge tube (1.5 mL), incubated at 38 °C to evaporate the methanol, and analyzed via enzyme-linked immunosorbent assay (ELISA). No differences were observed in fin cortisol levels among species or in fin and jawbone cortisol levels between washing solvents. The results of this study demonstrate that the sturgeon jawbone matrix is a promising alternative stress indicator to solid matrices.