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This study examined the effects of 7 days of heat stress on eight early lactating Holstein cows in climate-controlled chambers. The early lactating Holstein cows (42 ± 2 days in milk, 29.27 ± 0.38 kg/day milk yield, 1.21 ± 0.05 parity) were subjected to two 14-day periods, each consisting of 7 days of adaptation and 7 days of heat stress. Conditions were set to 22 °C and 50% humidity during adaptation, followed by heat stress periods with low-temperature, low-humidity (LTLH, 71 THI) and high-temperature, high-humidity (HTHH, 86 THI) treatments. Data from the last 7 days were analyzed using a mixed procedure in SAS. In the study, the HTHH group displayed marked physiological and biochemical changes on 14 days of heat stress exposure compared to the LTLH group. Firstly, the HTHH group's dry matter intake decreased by approximately 12% while their water intake increased by about 23%. Secondly, both milk yield and milk protein production in the HTHH group decreased by 10% and 20%, respectively. Thirdly, there was a reduction in white blood cells, hemoglobin, mean corpuscular hemoglobin, and platelets in the HTHH group, with concurrent increases in glucose, non-esterified fatty acids, and albumin concentrations. Additionally, the HTHH group exhibited elevated plasma concentrations of cortisol and haptoglobin. Moreover, the gene expression of heat shock protein 70 and heat shock protein 90 was significantly upregulated in the HTHH group's peripheral blood mononuclear cells. Lastly, key physiological indicators such as rectal temperature, heart rate, and skin temperature showed substantial elevations in the HTHH group. Considering the enormous negative effects observed in the analyzed blood metabolites, milk yield and compositions, and heat shock protein gene expression, early lactating Holstein cows were found to be more vulnerable to HTHH than LTLH over a 7 days exposure to heat stress., Competing Interests: Declaration of competing interest Authors certify that there is no conflict of interest with any financial organization regarding the material discussed in the manuscript., (Copyright © 2024. Published by Elsevier Ltd.)

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., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2024 Kim et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

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.

To find novel targeting moieties to lactating mammary gland, in vivo phage display screening was conducted with lactating rats and a peptide ligand, CLHQHNQMC (designated as MG1), which specifically homes to the mammary tissue during lactation, was identified through the consecutive in vivo biopannings. MG1 peptide ligand showed specific binding affinity to lactating mammary tissue without any preference to other organs tested in ex vivo and in vivo validation, and microscopy analysis revealed that systemically introduced MG1 could be specifically localized in the lactating mammary gland associated with mammary epithelia and alveolar vasculature. Based on the observation that binding of MG1-encoding phage to lactating mammary gland was competitively inhibited by synthetic MG1 peptide ligand, we attempted to identify a counterpart molecule corresponding to specific recognition of the MG1 and the acidic Ribosomal Protein Large P0 (RPLP0) was selected as a candidate receptor for MG1 by peptide affinity pull-down assay with protein extracts from lactating mammary tissue. We demonstrated specific expression of RPLP0 in mammary tissue, especially during lactation, by immunoblotting assays and also demonstrated that MG1 peptide ligand could be bound to, and internalized into, the cells effectively via specific interaction with RPLP0 by analysis using an in vitro endothelial cell model. The overall results suggest that the MG1 has a specific affinity with RPLP0 which are dominantly expressed on the mammary vasculature during lactation and this specific affinity enables the MG1 would be served as an effective homing ligand to deliver functional molecules to the lactating mammary gland., (Copyright © 2010 Elsevier Inc. All rights reserved.)

This study was conducted to investigate the amount of CLA synthesized endogenously by rat mammary tissues in response to TVA (a precursor for cis-9, trans-11 CLA endogenous synthesis) treatment as well as the differences in the protein expression of genes encoding the biosynthesis of CLA in rat mammary tissue and mouse mammary gland epithelia cells (HC11). Treatment with TVA resulted in improved CLA productivity. Furthermore, 2-DE revealed two spots in samples of mammary tissues and one spot in samples of mammary gland epithelia cells (HC11) that were consistently altered in the TVA treatment groups when compared with the control group (non-fatty acid). The mRNA expression patterns of three of the proteins (PDI, PRDX2, LAMR1), as measured by real-time PCR, were similar to the pattern of protein abundance. In addition, the expression of SCD mRNA in the mammary tissue of rats and HC11 cell treated with TVA was higher than in the control group. Our results suggest that the identified proteins may be related to CLA biosynthesis in mammary tissue., (Crown Copyright 2009. Published by Elsevier B.V. All rights reserved.)

Simple Summary: The composition of milk not only has nutritional implications, but is also directly related to the income of dairy producers. As regards milk's composition, concerns around milk protein have emerged from the increased consumption of casein products. The synthesis of proteins in milk is a highly complex and high-cost process, because the conversion efficiency of dietary protein to milk protein is very low in dairy cows. Thus, some studies have increased milk protein by using protein supplements or a single amino acid (AA) supply. AAs are the building blocks of protein, and can also stimulate the protein synthetic pathway. This review mainly concerns the use of AAs for producing milk protein in high-producing dairy cows, particularly with methionine, lysine, and histidine. Understanding the mechanisms of AAs will help to promote milk protein synthesis in the dairy industry. As the preference of consumers for casein products has increased, the protein content of milk from dairy cows is drawing more attention. Protein synthesis in the milk of dairy cows requires a proper supply of dietary protein. High protein supplementation may help to produce more milk protein, but residues in feces and urine cause environmental pollution and increase production costs. As such, previous studies have focused on protein supplements and amino acid (AA) supply. This review concerns AA nutrition for enhancing milk protein in dairy cows, and mainly focuses on three AAs: methionine, lysine, and histidine. AA supplementation for promoting protein synthesis is related to the mammalian target of rapamycin (mTOR) complex and its downstream pathways. Each AA has different stimulating effects on the mTOR translation initiation pathway, and thus manifests different milk protein yields. This review will expand our understanding of AA nutrition and the involved pathways in relation to the synthesis of milk protein in dairy cows. [ABSTRACT FROM AUTHOR]

Simple Summary: This study examined the effect of acetate and L-tryptophan-conjugated bypass amino acid (ACT), supplemented (15 g/day) to Holstein cows during late pregnancy, on their productivity and the performance of offspring. We identified that the supplementation of ACT incorporated into diet was beneficial for improving the feed intake, blood hematology, and metabolites of the prepartum, and also had a positive effect on reducing saturated fatty acids in the colostrum of the cows postpartum and on the body weight of the newborn calves. The results of this study suggest that ACT supplementation improves the productivity of dairy cows. 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. [ABSTRACT FROM AUTHOR]

Simple Summary: In this study, we characterize the influence of short-term (4 days) heat stress on Holstein cows during early lactation. The use of indicators, such as production performance, physiological variables, blood parameters, micro-RNA expression, and metabolomes, in heat-stressed cows during early lactation—which is a high-stress phase—may provide insights into how to deal with the level of damage to dairy cows, through appropriate nutritional and management strategies. We identify that short-term heat stress has a negative effect, to some extent, on feed and water intake, rectal temperature, heart rate, blood hematology and metabolites, milk characteristics, miRNA expression in milk, and metabolomics in blood. 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. [ABSTRACT FROM AUTHOR]

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. β-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 β-casein synthesis by stimulating genes, proteins, and pathways related to protein and energy metabolism. [ABSTRACT FROM AUTHOR]