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The effect of different sub-pasteurization heat treatments and different ripening times was investigated in this work. The metabolite profiles of 95 cheese samples were analyzed using GC-MS in order to determine the effects of thermal treatment (raw milk, 57 °C and 68 °C milk thermization) and ripening time (105 and 180 days). ANOVA test on GC-MS peaks complemented with false discovery rate correction was employed to identify the compounds whose levels significantly varied over different ripening times and thermal treatments. The univariate t-test classifier and Partial Least Square Discriminant Analysis (PLS-DA) provided acceptable classification results, with an overall accuracy in cross-validation of 76% for the univariate model and 72% from the PLS-DA. The metabolites that mostly changed with ripening time were amino acids and one endocannabinoid (i.e., arachidonoyl amide), while compounds belonging to the classes of biogenic amines and saccharides resulted in being strongly affected by the thermization process., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Whey proteins are N-glycosylated proteins that play important roles in a variety of biological processes including immune defense. However, the N-glycosylation of yak colostrum (YC) and mature milk (YM) whey proteins is unknown. Therefore, this study systematically compared and analyzed YC and YM whey N-glycoproteomes using the 4D label-free technique. We identified 162 glycoproteins, 222 glycosylated peptides, and 234 glycosylation sites in YC and YM, of which 59 glycoproteins were differentially expressed in YC and YM. According to gene ontology annotation and KEGG pathway metabolism analysis, the differentially expressed N-glycoproteins were highly enriched in "cell adhesion", "extracellular region", and "calcium binding", and were mainly involved in the extracellular matrix (ECM)-receptor interaction pathway. The immunity-related N-glycoproteins, such as platelet glycoprotein 4 (CD36) and polymeric immunoglobulin receptor (PIGR), were observed to be different between YC and YM. The results revealed the glycosylation sites, composition, and biological functions of YC and YM whey N-glycoprotein, which supplemented our understanding of the N-glycosylation of yak whey proteins.

Goat milk is rich in a variety of nutrients that are important for intestinal health and disease prevention. However, the role of exosomes in goat milk remains to be elucidated. This study investigated for the first time the therapeutic efficacy and molecular underlying mechanisms of mature milk exosomes (M-exo) and goat colostrum exosomes (C-exo) on dextran sodium sulfate-induced ulcerative colitis (UC) in mice. The findings demonstrate that M-exo and C-exo significantly improved physiological indices, suppressed the secretion of proinflammatory cytokines, and diminished oxidative stress and apoptosis in UC mice. Moreover, C-exo and M-exo restored the intestinal barrier function, remodeled the gut microbiota, and improved metabolite composition in the feces of colitis mice. In conclusion, goat milk exosomes ameliorate UC in mice, which provides a basis for the development of functional food applications for the prevention and treatment of inflammatory bowel disease.

Goat milk is abundant in nutrients, particularly in milk fats, which confer health benefits to humans. Exploring the regulatory mechanism of fatty acid synthesis is highly important to understand milk composition manipulation. In this study, we used chromatin immunoprecipitation sequencing (ChIP-seq) on goat mammary glands at different lactation stages which revealed a novel lactation regulatory factor: cell death-inducing DFFA-like effector B (CIDEB). RT-qPCR results revealed that CIDEB was significantly upregulated during lactation in dairy goats. CIDEB overexpression significantly increased the expression levels of genes involved in fatty acid synthesis ( ACACA , SCD1 , p < 0.05; ELOVL6 , p < 0.01), lipid droplet formation ( XDH , p < 0.05), and triacylglycerol (TAG) synthesis ( DGAT1 , p < 0.05; GPAM , p < 0.01) in goat mammary epithelial cells (GMECs). The contents of lipid droplets, TAG, and cholesterol were increased ( p < 0.05) in CIDEB-overexpressing GMECs, and knockdown of CIDEB led to the opposite results. In addition, CIDEB knockdown significantly decreased the proportion of C16:0 and total C18:2. Luciferase reporter assays indicated that X-box binding protein 1 (XBP1) promoted CIDEB transcription via XBP1 binding sites located in the CIDEB promoter. Furthermore, CIDEB knockdown attenuated the stimulatory effect of XBP1 on lipid droplet accumulation. Collectively, these findings elucidate the critical regulatory roles of CIDEB in milk fat synthesis, thus providing new insights into improving the quality of goat milk.

Background: Goat milk has gained global attention for its unique nutritional properties and potential health benefits. Advancements in functional genomic technologies have significantly progressed genetic research on milk composition traits in dairy goats., Results: This review summarizes various research methodologies applied in this field. Genome-wide association studies (GWAS) have identified genomic regions associated with major milk components, with the diacylglycerol acyltransferase 1 ( DGAT1 ) gene and casein gene cluster consistently linked to milk composition traits. Transcriptomics has revealed gene expression patterns in mammary tissue across lactation stages, while the role of non-coding RNAs (such as miRNAs and circRNAs) in regulating milk composition has been confirmed. Proteomic and metabolomic studies have not only helped us gain a more comprehensive understanding of goat milk composition characteristics but have also provided crucial support for the functional validation of genes related to milk components. The integration of multi-omics data has emerged as an effective strategy for elucidating complex regulatory networks from a systems biology perspective., Conclusions: Despite progress, challenges remain, including refining reference genomes, collecting large-scale phenotypic data, and conducting functional validations. Future research should focus on improving reference genomes, expanding study populations, investigating functional milk components, exploring epigenetic regulation and non-coding RNAs, and studying microbiome-host genome interactions. These efforts will inform more precise genomic and marker-assisted selection strategies, advancing genetic improvements in milk composition traits in dairy goats.

Background: Milk clotting enzymes, essential for milk coagulation in cheese production, are obtained from the stomach of young ruminants, an expensive and limited source. This study was accomplished by finding a suitable alternative. Bacterial isolates recovered from honey were screened for milk clotting enzyme activity. and further, by immobilization of the microorganisms to enhance stability and facilitate their repeated use., Result: The most effective enzyme was produced by a microbe identified as Bacillus amyloliquefaciens based on 16 S rRNA sequencing. The cells were encapsulated in Ca 2+ alginate beads. These beads retained complete enzyme production after being used five times. Glucose and Soybean were selected as the most favorable carbon and nitrogen sources, respectively. The optimum temperature for activity was 35 ℃ for both free and immobilized cells but as the temperature was increased to 55 °C and above, the encapsulated form retained more activity than the free cells. The pH optimum shifted from 6.5 to 7 for the free cells to 7-7.5 for the immobilized cells. The immobilization process decreased the activation energy for enzyme production and activity, prolonged the enzyme half-life, and increased the deactivation energy. Enzyme produced by immobilized cells generated a more compact cheese., Conclusions: The finding of this study was to identify a less expensive source of milk-clotting enzymes and confirm the success of cell immobilization in improving cell rigidity and stability. Also, immobilization of this B. amyloliquefaciens strain offers an enzyme source of value for industrial production of cheese., (© 2024. The Author(s).)

The impact of nutritional modification to increase functional polyunsaturated fatty acids (PUFA), such as n-3 and n-6 fatty acids (FA) or conjugated linoleic acid (CLA), on milk proteome profile during early lactation remains largely unknown. We used an untargeted proteomics approach to investigate the impact of lactation day and PUFA supplementation on the proteome signature in skimmed milk over the course of early lactation. Sixteen Holstein dairy cows received abomasal infusion of saturated FA (CTRL) or a mixture of essential FA and CLA (EFA + CLA group) from - 63 to + 63 days relative to parturition. Using quantitative proteomics, 479 unique proteins were identified in skimmed milk at days 1, 28, and 63 postpartum. The top discriminating proteins between transition milk (day 1) and mature milk (days 28 and 63), including members of complements (i.e. C2 and C5), growth factor (TGFB2), lipoproteins (i.e. APOE and APOD), and chaperones (i.e. ST13 and CLU), are associated with calves' immune system and gut development. The EFA + CLA supplementation moderately affected a few proteins associated with regulating mammary glands' lipogenesis through the (re)assembly of lipoprotein particles, possibly under the PPAR signaling pathway. Collectively, skimmed milk proteome is dynamically regulated initially by cow's metabolic and physiological changes and to a lesser extent by nutritional PUFA modifications., (© 2024. The Author(s).)

Dietary supplementation of fat can be an important source of energy to compensate for the reduction in dry matter intake in dairy cows during heat stress periods. Studies have reported that supplementing dairy cow diets with linseed oil (LO) can increase milk yield and enhance the levels of beneficial fatty acids, such as omega-3 fatty acids, in the milk. The objective of this research was to evaluate the effect of LO supplementation on milk fatty acids profile, milk yield and composition, and physiological parameters of grazing cows. The study was conducted in two seasons, one in spring and one in summer. A 2 × 2 Latin square design was used in each experiment. Twelve Holstein and crossbred Holstein x Jersey cows were involved in each season. Cows were divided into two groups: control (TC) with no supplementation and treatment (TL) supplemented with 400 g/day of LO. The results showed that LO supplementation altered the milk fatty acid profile: decreased concentrations of short and medium-chain fatty acids (C10:0 - C17:1) except for C13:0 and increased concentrations of long-chain fatty acids (C18, C18:1 (both trans and cis isomers), C18:2 (specific conjugated linoleic acid - CLA isomers), and C18:3 n3 (omega-3)). Additionally, milk yield increased by 1.5 l per day during summer in LO-supplemented cows, while milk fat, protein, and casein content decreased. Milk stability increased by 2.2% in the LO-supplemented group. LO-supplemented cows reduced internal body temperature and heart frequency in the afternoon and increased daily rumination time by 20 min. In conclusion, LO supplementation can be an effective strategy to improve the nutritional profile of milk by altering fatty acid composition towards potentially healthier fats, mitigate the negative effects of heat stress on grazing cows during summer, as evidenced by reduced body temperature and heart frequency and increase milk yield., (© 2024. The Author(s), under exclusive licence to Springer Nature B.V.)

Milk-derived extracellular vesicles (mEVs) are beneficial to the health of infants. However, the effect of mEVs on early intestinal inflammation is not well established. Herein, weaned colitic mice were used to explore the potential effects and underlying mechanisms of porcine mEVs (pmEVs) on intestinal inflammation during early life. We found that pmEVs administration attenuated early life intestinal inflammation and promoted colonic barrier integrity in mice. The anti-inflammatory effect of pmEVs was achieved by shifting a proinflammatory macrophage (M1) toward an anti-inflammatory macrophage (M2). Moreover, pmEVs can be absorbed by macrophages and reduce proinflammatory polarization (stimulated by LPS) in vitro. Noteworthily, ssc-let- 7 c was found to be highly expressed in pmEVs that can regulate the polarization of macrophages by targeting the tensin homologue deleted on chromosome ten (PTEN), thereby activating the PI3K/Akt pathway. Collectively, our findings revealed a crucial role of mEVs in early intestinal immunity and its underlying mechanism.

Tissue-specific deficiency of nicotinamide phosphoribosyl transferase (NAMPT), the rate-limiting enzyme of the nicotinamide adenine dinucleotide (NAD + )-salvage pathway, causes a decrease of NAD + in the tissue, resulting in functional abnormalities. The NAD + -salvage pathway is drastically activated in the mammary gland during lactation, but the significance of this has not been established. To investigate the impact of NAD + perturbation in the mammary gland, we generated two new lines of mammary gland epithelial-cell-specific Nampt-knockout mice (MGKO). LC-MS/MS analyses confirmed that the levels of NAD + and its precursor nicotinamide mononucleotide (NMN) were significantly increased in lactating mammary glands. We found that murine milk contained a remarkably high level of NMN. MGKO exhibited a significant decrease in tissue NAD + and milk NMN levels in the mammary gland during lactation periods. Despite the decline in NAD + levels, the mammary glands of MGKO appeared to develop normally. Transcriptome analysis revealed that the gene profiles of MGKO were indistinguishable from those of their wild-type counterparts, except for Nampt. Although the NMN levels in milk from MGKO were decreased, the metabolomic profile of milk was otherwise unaltered. The mammary gland also contains adipocytes, but adipocyte-specific deficiency of Nampt did not affect mammary gland NAD +  metabolism or mammary gland development. These results demonstrate that the NAD +  -salvage pathway is activated in mammary epithelial cells during lactation and suggest that this activation is required for production of milk NMN rather than mammary gland development. Our MGKO mice could be a suitable model for exploring the potential roles of NMN in milk., Competing Interests: Declaration of competing interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests:Yo-ichi Nabeshima reports financial support was provided by Meiji Holdings Co Ltd. Kouya Hattori, Rika Azuma-Suzuki, Seiichiro Higashi, Takashige Hamaguchi, Yoshie Saito, Masashi Morifuji reports a relationship with Meiji Holdings Co Ltd that includes: employment. The Department of Aging Science and Medicine is endowed by Meiji Holdings Co., Ltd. If there are other authors, they declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Inc. All rights reserved.)

Inflammatory diseases of the intestinal tract in piglets severely impair the economic performance of pig farms. Pig milk exosomes can encapsulate miRNAs which can then enter the piglet intestine to play an immunomodulatory role. Previously, we comparatively analyzed and identified exosomal miRNAs in the colostrum and mature milk of Bamei and Landrace pigs, and we screened for ssc-miR-22-3p, which is associated with inflammation and immune response; however, the role played by ssc-miR-22-3p in the immune response in IPEC-J2 cells is not yet clear. In this study, we first constructed a pig intestinal inflammatory response model using Lipopolysaccharide (LPS) and Polyinosinic-polycytidylic acid (Poly (I:C)), and we investigated the role of ssc-miR-22-3p targeting MAPK14 in the regulation of LPS and Poly (I:C)-induced inflammatory injury in IPEC-J2 cells by RT-qPCR, cell counting kit-8 (CCK-8), EdU staining, lactate dehydrogenase (LDH) activity assay, and dual luciferase reporter gene assay. We successfully established LPS and Poly (I:C)-induced cell damage models in IPEC-J2 cells. The immune response of IPEC-J2 cells was stimulated by induction of IPEC-J2 cells at 10 μg/mL LPS and 20 μg/mL Poly (I:C) for 24 h. Overexpression of ssc-miR-22-3p decreased cytokine expression and promoted cell viability and proliferation. The functional enrichment analysis revealed that ssc-miR-22-3p targets genes enriched in the pathways of negative regulation of inflammatory response and bacterial invasion of epithelial cells. The validity of the binding site of ssc-miR-22-3p to MAPK14 was tested by a dual luciferase reporter gene. Pig milk exosome ssc-miR-22-3p promotes cell viability and proliferation by targeting MAPK14 , and it alleviates LPS and Poly (I:C)-induced inflammatory responses in IPEC-J2 cells.

On the basis of comparisons between bovine and ovine genome mapping information, the aim of the study was to analyze the genetic diversity of selected DNA microsatellites from the bovine genome and to investigate their correlation with the average daily milk yield in Awassi sheep. 18 informative microsatellite markers were selected from the significant QTL regions affecting milk yield identified in the bovine genome in previous studies. The selected microsatellite markers were then amplified by PCR as reciprocal amplifications on the genomic DNA of Awassi sheep, with standard daily milk yield records. Thus, in this study, 18 microsatellite markers associated with milk yield in the bovine genome were examined for both determination of genetic polymorphism within the flock and the effects of marker loci on average daily milk yield in Awassi sheep. Allele frequencies of markers were determined based on the results of fragment analysis. The analysis of variance showed that the 123 bp allele at the marker locus BMS1341 on BTA2 significantly influenced the average daily milk yield of Ivesi sheep (P < 0.01). On the other hand, the BMS381 locus with a 115 bp allele on BTA2, the MCM140 locus with a 185 bp allele on BTA6, the BMS2721 locus with a 155 bp allele, the BM1237 locus with 174 and 180 bp alleles on BTA7, and finally, the BMS1967 locus with a 117 bp allele, the BM4208 locus with 176 and 182 bp alleles, and the INRA locus with a185 bp allele on BTA8 showed moderately significant effects on the average daily milk yield of Ivesi ewes (P < 0.05)., (© 2024. The Author(s), under exclusive licence to Springer Nature B.V.)

This study systematically examines the patterns of milk yield (MY, kg), fat (FAT, %), and protein (PROT, %) in a diverse population of Thai multibreed dairy cattle, considering the tropical environment's impact on lactating cows. Using a dataset of 47,205 monthly test-day records from 4,440 first-lactation cows across 446 farms, we analyze variations and interrelationships through mathematical averaging and introduce the fat-to-protein ratio (FPR) to assess acidosis (FPR < 1.1) and ketosis (FPR > 1.5) risks during lactation. Pearson correlation analysis elucidated trait associations. The findings, aligned with established lactation norms, indicate peak production at 297 days in milk (DIM) for FAT (4.08%; SD = 0.96%), PROT (3.43%; SD = 0.47%), and 52 DIM for MY (18.09 kg; SD = 4.91 kg). Nadirs are observed at 72 DIM for FAT (3.27%; SD = 0.74%), 47 DIM for PROT (2.86%; SD = 0.36%), and 299 DIM for MY (9.05 kg; SD = 2.95 kg). FPR variations highlight acidosis (46.48%), normal (43.66%), and ketosis (9.86%), especially during early lactation (100 DIM). Significant negative correlations emerge between MY, FAT, and PROT (P < 0.05), while a positive correlation is identified between FAT and PROT (P < 0.01), with robust correlations during early lactation. This study contributes to understanding tailored nutritional strategies for dairy cows' holistic health and sustainability in tropical environments, guiding efficient production practices and mitigating health-related productivity impediments., (© 2024. The Author(s), under exclusive licence to Springer Nature B.V.)

Exosomes are cell-derived, membrane-surrounded particles that deliver bioactive molecules to various cells. Due to their small size, low immunogenicity, extended blood circulation, and involvement in cellular communication, they hold potential as effective drug carriers. Exosomes are present in various biological fluids, including mare's milk, a traditional drink in Central Asia. This study aims to compare exosome isolation methodologies and determine the stability of mare's milk-derived exosomes as potential therapeutic carriers. Three extraction methods-immunoprecipitation, size exclusion chromatography, and total exosome isolation-were compared in terms of exosome characteristics, purity, and content. The isolated exosomes were then loaded with quercetin, and their ability to increase its bioavailability was tested in vitro and in vivo. Total exosome isolation was identified as the most efficient method for producing high-quality exosomes. These exosomes were loaded with quercetin and compared to free quercetin and exosomes alone. Exosomes loaded with 80 µM quercetin significantly restored β-galactosidase activity and cellular viability in doxorubicin-treated cells, exhibiting similar potency to 160 µM free quercetin. In aged model animals, treatment with quercetin-loaded exosomes resulted in significantly less acute and subacute damage to the myocardium, kidneys, and liver compared to untreated control animals. This study provides a proof-of-concept that mare's milk-derived exosomes can be effectively absorbed by cells and animal tissues, supporting their potential use as drug carriers.

Genome-wide association studies (GWASs) are used to identify quantitative trait loci for phenotypic traits of interest. The use of multilocus mixed models allows to correct for population stratification and account for long-range linkage disequilibrium. In this study, GWASs were conducted to identify the genetic bases of milk production (milk yield, protein and fat composition, and yield) in two autochthonous dual-purpose cattle breeds from the Aosta Valley. Using either the breeding values or the deregressed proofs, common significative single nucleotide polymorphisms have been identified for milk yield, protein percentage, and fat percentage. Two major quantitative trait loci regions have been identified on the chromosomes 5 and 14 for the fat percentage, harbouring the MGST1, CYHR1, VPS28, and CPSF1 genes. For the protein percentage, a candidate region has been identified on BTA 6; in this region, the CSN1S1, CSN2, HSTN, CSN3, and RUFY3 genes are annotated. Most of the identified genes have already been associated with milk composition in other studies on cosmopolitan and local cattle. These results show that the genes involved in milk composition quantitative traits in the Aosta cattle are common also in other cattle breeds and they can be further investigated with the use of whole genome sequencing data., (Copyright © 2024 The Authors. Published by Elsevier B.V. All rights reserved.)

A meta-analysis was performed to investigate the effects of capsaicin (CAP) on lactational performance and blood chemistry and cell counts in dairy cattle. The database comprised 11 peer-reviewed studies published between 2011 and 2024. The overall effect of CAP, challenge (e.g., ketosis, glucose tolerance, LPS), and dose were considered explanatory variables in the analysis. Lactation performance response variables included DMI, milk yield (MY), ECM yield, feed efficiency (FE), milk components, and BW. Hematological response variables included red and white blood cell counts and blood BHB, total fatty acids, insulin, and glucose concentrations. Data were analyzed using random- and mixed-effect models in the "robumeta" package in RStudio. Milk yield was increased by 2.9% by CAP when compared with control. Capsaicin supplementation increased FE by 3.4% compared with control. Milk fat concentration and yield were also increased by CAP compared with control by 2.6% and 4.0%, respectively. Blood glucose concentrations were decreased 2.5% by CAP supplementation, whereas insulin levels were unaffected. Cows fed CAP during a challenge had higher MY and FE and tended to have lower blood glucose than their control counterparts. Overall, this analysis suggests that CAP supplementation may be directly affecting host physiology by altering glucose metabolism, but further research to define the mechanism is warranted., (The Authors. Published by Elsevier Inc. on behalf of the American Dairy Science Association®. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).)

Nitrogen (N) utilisation efficiency (NUE, milk N yield [g/d]/N intake [g/d]) is an important performance indicator in dairy farming. Determining the NUE-associated blood metabolite profile will contribute to the optimisation of nutritional strategies to further improve NUE among dairy cows. Here, 20 primiparous lactating cows with days in milk ranging from 95 to 115 days were selected from a total of 1 221 cows. Each cow's N intake and milk N yield were measured for 7 days. Subsequently, blood samples were collected before morning feeding. Based on analysis and calculations, cows were retrospectively classified into two groups based on their NUE values, namely, a low NUE group (LNUE, NUE = 24.8 ± 1.6%, n = 10, mean ± SD) and a high NUE group (HNUE, NUE = 35.2 ± 1.7%, n = 10, mean ± SD). Plasma samples were selected from six cows in each group for metabolomics and amino acid profiling. Among the 41 differential metabolites (DMs) identified in the metabolomic analysis, sucrose, MG(0:0/22:1(13Z)/0:0), 2-amino-6-hydroxyhexanoic acid, and L-glutamine exhibited significant correlations with NUE, milk yield, and BW (P < 0.05). Moreover, the five differential amino acids and amino acid metabolites (DAAs) identified in the amino acid profiling and 5 of the 6 differential amino acids and amino acid conjugates identified by plasma metabolomics were found to be less abundant in the HNUE group (P < 0.05). Specifically, there was a 39.4% decrease in L-arginine content and a 29.2% decrease in L-glutamine content (P < 0.05). Pathway analysis indicated that the DMs and DAAs were mainly involved in arginine biosynthesis, glutathione metabolism, arginine and proline metabolism, and tryptophan metabolism (pathway impact > 0.1). These results provided new insights into the new blood metabolite profile associated with NUE in dairy cows. These new insights can provide foundational information for the formulation of new strategies to further enhance NUE in dairy cows., (Copyright © 2024 The Author(s). Published by Elsevier B.V. All rights reserved.)

Dairy cows experiencing heat stress (HS) during the precalving portion of the transition period give birth to smaller calves and produce less milk and milk protein. Supplementation of rumen-protected methionine (RPM) has been shown to modulate protein, energy, and placenta metabolism, making it a potential candidate to ameliorate HS effects. We investigated the effects of supplementing RPM to transition cows under HS induced by electric heat blanket (EHB) on cow-calf performance. Six weeks before expected calving, 53 Holstein cows were housed in a tiestall barn and fed a control diet (CON, 2.2% Met of MP) or a CON diet supplemented with SmartamineM (MET, 2.6% Met of MP, Adisseo Inc., France). Four weeks precalving, all MET and half CON cows were fitted with an EHB. The other half of the CON cows were considered thermoneutral (TN), resulting in 3 treatments: CONTN (n = 19), CONHS (n = 17), and METHS (n = 17). Respiratory rate (RR), skin temperature (ST), and rectal temperature (RT) were measured thrice weekly and core body temperatures recorded biweekly. Postcalving BW and BCS were recorded weekly, and DMI was calculated and averaged weekly. Milk yield was recorded daily and milk components were analyzed every third DIM. Biweekly AA and weekly nonesterified fatty acids (NEFA), BHB, insulin, and glucose were measured from plasma. Calf birth weight and 24 h growth, thermoregulation, and hematology profile were measured and apparent efficiency of absorption (AEA) of immunoglobulins was calculated. Data were analyzed using the MIXED procedure of SAS with 2 preplanned orthogonal contrasts: CONTN versus the average of CONHS and METHS (C1) and CONHS versus METHS (C2). Relative to TN, EHB cows had increased RT during the postcalving weeks and increased RR and ST during the entire transition period. Body weight, BCS, DMI, and milk yield were not affected by the EHB or RPM. However, protein percentage and SNF were lower in CONHS, relative to METHS cows. At calving, METHS dams had higher glucose concentrations, relative to CONHS, and during the postcalving weeks, the EHB cows had lower NEFA concentrations than TN cows. Calf birthweight and AEA were reduced by HS, whereas RR was increased by HS. Calf withers height tended to be shorter and RT were lower in CONHS, compared with METHS heifers. Overall, RPM supplementation to transition cows reverts the negative effect of HS on blood glucose concentration at calving and milk protein percentage in the dams and increases wither height while decreasing RT in the calf., (The Authors. Published by Elsevier Inc. on behalf of the American Dairy Science Association®. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).)

Early lactation is a critical period for dairy cows, as energy requirements rapidly increase with the onset of lactation; however, early-lactation DMI in pasture-based systems are under measured. The objectives of this study were (1) to measure and profile total DMI (TDMI) and animal performance of dairy cows during early lactation in a pasture-based system, (2) to investigate early-lactation energy balance in pasture-based systems, and (3) to examine production efficiencies, including TDMI and milk solids production per 100 kg of BW. Eighty spring-calving dairy cows were allocated to a grazing group as they calved over a 2-yr period (2021 and 2022). Cows were offered a daily herbage allowance to achieve a postgrazing sward height of 4 cm, with silage supplementation when necessary due to inclement weather. Total DMI was measured using the n-alkane technique over a 12-wk period from February 1, 2021, to April 23, 2022. Total DMI and daily milk yield were significantly affected by parity with both variables being greatest for third-parity animals (17.7 kg of DM and 26.3 kg/cow per day, respectively), lowest for first parity (13.2 kg of DM and 19.6 kg/cow per day, respectively) and intermediate for second-parity animals (16.8 kg of DM and 24.1 kg/cow per day, respectively). Peak TDMI was reached on wk 10 for first-parity animals (14.6 kg of DM), wk 11 for second parity animals (19.3 kg of DM) and wk 12 for third-parity animals (19.9 kg of DM). Parity also had a significant effect on unité fouragère lait (UFL; feed units for milk) feed balance as first-parity animals experienced a greater degree of negative energy balance (-3.2 UFL) compared with second- and third-parity animals (-2.3 UFL). Breed and parity had an effect on production efficiencies during the first 12 wk of lactation as Jersey × Holstein Friesian cows had greater TDMI/100 kg of BW and milk solids/100 kg of BW compared with Holstein Friesian cows., (The Authors. Published by Elsevier Inc. on behalf of the American Dairy Science Association®. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).)

Determination of energy requirements for growth depends on measuring the composition of BW gain. Previous studies have shown that the composition of gain can be altered in young dairy calves by the composition of the milk replacer diet. Here, our objective was to determine body composition and the composition of empty body gain in young calves fed increasing amounts of a milk replacer containing adequate CP. Male Holstein calves underwent an adjustment period of 14 d after birth in which they were fed whole-waste milk at 10% of BW. Calves were then stratified by BW and randomly assigned to either an initial slaughter group (n = 11) or to groups fed 1 of 3 milk replacer amounts and slaughtered after 35 d of growth. Calves in all treatments consumed the same milk replacer containing 24.8% CP (DM basis; from all milk proteins) and 18.9% fat, reconstituted to 12.5% solids. Treatments were milk replacer fed at 1.25% of BW (DM basis; n = 6), 1.75% of BW (n = 6), or 2.25% of BW (n = 8), adjusted weekly as calves grew. Calves fed at 1.25% or 1.75% of BW were fed twice daily and those fed 2.25% of BW were fed 3 times daily. No starter was offered. Postslaughter, the bodies of calves were separated into 4 fractions: carcass; total viscera minus digesta; head, hide, feet, and tail;, and blood. The sum of those 4 fractions was empty BW (EBW), which increased linearly as the amount of milk replacer increased. Final heart girth and body length, but not withers height, increased linearly as intake increased. Gain:feed increased linearly with increasing milk replacer. Feeding more milk replacer increased the amounts of lean tissue and fat in the body. The percentages of water and protein in the final body decreased linearly, whereas fat percentage and energy content increased linearly as intake increased. As gain increased, the percentage of protein in gain decreased and the percentage of fat increased, resulting in an increase of energy content of EBW gain. Efficiency of energy use (retained energy:gross energy intake) increased linearly but retained energy:ME available for growth was not different among treatments. Efficiency of protein use increased quadratically as feeding rate increased; there was no further increase at 2.25% of BW. Plasma insulin-like growth factor 1, insulin, and glucose increased linearly, whereas urea-N decreased linearly, as milk replacer intake increased. Our data document changes in body composition that affect estimates of retained energy in the bodies of calves slaughtered at a common age. These data are important for calculations of energy requirements for young calves., (The Authors. Published by Elsevier Inc. on behalf of the American Dairy Science Association®. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).)

The study investigated the effects of dietary protein level and the inclusion of hydroponic barley sprouts (HB) on lactation performance, blood biochemistry and N use efficiency in mid-lactation dairy cows. Treatments were arranged in a 2 × 2 factorial design with 2 CP levels (16.8% and 15.5% of DM), with HB (4.8% of DM, replacing 4.3% of alfalfa hay and 0.5% of distillers dried grains with solubles [DDGS]) or without HB. Forty-eight multiparous Holstein dairy cows (146 ± 15 DIM, 40 ± 5 kg/d of milk) were randomly allocated to 1 of 4 diets: high-protein diet (16.8% CP, HP), HP diet with HB (HP+HB), low-protein diet (15.5% CP, LP), or LP diet with HB (LP+HB). An interaction between CP × HB on DMI was detected, with DMI being unaffected by HB inclusion in cows fed the high-protein diets, but was lower in cows fed HB when the low-protein diet was fed. A CP × HB interaction was also observed on milk and milk protein yield, which was higher in cows fed HB with HP, but not LP. Inclusion of HB also tended to reduce milk fat content, and feeding HP resulted in a higher milk protein and MUN content, but lower milk lactose content. Feed efficiency was increased by feeding HP or HB diets, whereas N use efficiency was higher for cows fed LP or HB diets. There was an interaction on the apparent total-tract digestibility of DM and CP, which was higher when HB was fed along with HP, but reduced when fed with LP, whereas the digestibility of ADF was increased by feeding low-protein diets. In conclusion, feeding a low-protein diet had no adverse effect on cow performance, while feeding HB improved milk and milk component yield, and N efficiency when fed with a high-CP diet, but compromised cow performance with a low-CP diet., (The Authors. Published by Elsevier Inc. on behalf of the American Dairy Science Association®. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).)

The aim of this study was to evaluate the effects of different anti-mycotoxin feed additives on the concentration of mycotoxins in milk, urine, and blood plasma of dairy cows fed diets artificially contaminated with mycotoxins. Secondarily, performance, total-tract apparent digestibility of nutrients, and blood parameters were evaluated. Twelve multiparous cows (165 ± 45 DIM, 557 ± 49 kg BW, and 32.1 ± 4.57 kg/d milk yield at the start of the experiment) were blocked according to parity, milk yield, and DIM and used in a 4 × 4 Latin square design experiment with 21-d periods, where the last 7 d were used for sampling and data analysis. Treatments were (1) mycotoxin group (MTX), basal diet (BD) without anti-mycotoxin feed additives; (2) hydrated sodium calcium aluminosilicate (HSCA), HSCA added to the BD at 25 g/cow per day; (3) mycotoxin deactivator (MD; Mycofix Plus, dsm-firmenich) added to the BD at 15 g/cow per day (MD15); and (4) MD added to the BD at 30 g/cow per day (MD30). Cows from all treatments were challenged with a blend of mycotoxins containing 404 μg of aflatoxin B 1 , 5,025 μg of deoxynivalenol (DON), 8,046 μg of fumonisins (FUM), 195 μg of T2 toxin (T2), and 2,034 μg of zearalenone (ZEN) added daily to the BD during the last 7 d of each period. Neither performance (milk yield and composition) nor nutrient digestibility was affected by treatments. All additives reduced aflatoxin M 1 (AFM1) concentration in milk, whereas MD15 and MD30 group had lower excretion of AFM1 in milk than HSCA. Deoxynivalenol, FUM, T2, or ZEN were not detected in milk of MD15 and MD30. Concentrations in milk of DON, FUM, T2, and ZEN were similar between MTX and HSCA. Except for AFM1, none of the analyzed mycotoxins were detected in urine of MD30 group. Comparing HSCA to MD treatments, the concentration of AFM1 was greater for HSCA, whereas MD30 was more efficient at reducing AFM1 in urine than MD15. Aflatoxin M1, DON, FUM, and ZEN were not detected in the plasma of cows fed MD30, and DON was also not detected in MD15 group. Plasma concentration of FUM was lower for MD15, similar plasma FUM concentration was reported for HSCA and MTX. Plasma concentration of ZEN was lower for MD15 than MTX and HSCA. Serum concentrations of haptoglobin and hepatic enzymes were not affected by treatments. Blood concentration of sodium was lower in HSCA compared with MD15 and MD30 groups. In conclusion, the mycotoxin deactivator proved to be effective in reducing the secretion of mycotoxins in milk, urine, and blood plasma, regardless of the dosage. This reduction was achieved without adverse effects on milk production or total-tract digestibility in cows fed multi-mycotoxin-contaminated diets over a short-term period. Greater reductions in mycotoxin secretion were observed with full dose of MD., (The Authors. Published by Elsevier Inc. on behalf of the American Dairy Science Association®. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).)

Brown Swiss (BS) cows have greater urea concentrations in milk and blood compared with Holstein (HO) cows. We tested the hypothesis that BS and HO cows differ in kidney function and nitrogen excretion. Blood, saliva, urine, and feces were sampled in 31 multiparous BS and 46 HO cows kept under identical feeding and management conditions. Samples were collected at different lactational stages after the monthly DHIA control test-day. To test the glomerular filtration rate (GFR) and urea excretion, concentrations of creatinine and urea were measured in serum, urine, and saliva. As an additional marker to estimate GFR, we determined symmetric dimethylarginine (SDMA) in serum. Feces were analyzed for DM content and nitrogen concentration. Data on milk urea and protein concentrations and daily milk yield were obtained from the monthly DHIA test-day records. The effects of breed, time, and parity number on blood, saliva, urine, feces, and milk parameters were evaluated with the GLM procedure of SAS, with breed, time, and parity number as fixed effects. Differences between BS and HO were assessed by the Tukey-corrected t-test. Concentrations of urea, creatinine, and SDMA in serum, were greater in BS than in HO cows: 5.46 ± 0.19 versus 4.72 ± 0.13 mmol/L (urea), 105.96 ± 2.23 versus 93.07 ± 1.50 mmol/L (creatinine), and 16.78 ± 0.69 versus 13.39 ± 0.44 µg/dL (SDMA). We observed a greater urea concentration in BS cows (25.8 ± 0.7 vs. 21.8 ± 0.7 mg/dL) and protein content in milk (3.70% ± 0.08% vs. 3.45% ± 0.07%) than in HO cows. Urea and creatinine concentrations in urine and saliva did not differ between breeds. No differences between BS and HO were observed for milk yield, fecal DM, and fecal nitrogen content. Dry matter intake and BW were similar in BS and HO cows. Despite greater urea, creatinine, and SDMA concentrations in blood, as well as a higher milk urea content in BS compared with HO, respective concentrations in urine did not differ between breeds. In conclusion, our results demonstrate a lower renal GFR in BS compared with HO cows, thereby contributing to the greater plasma urea concentration in BS cows. However, estimation of nitrogen excretion via milk, urine, and feces does not entirely reflect nitrogen turnover within the animal., (The Authors. Published by Elsevier Inc. on behalf of the American Dairy Science Association®. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).)

Dietary carbohydrate manipulation can be used to reduce enteric CH 4 emission, but few studies have focused on the interaction of the different types of carbohydrates that can affect feed intake and ruminal fermentation. Understanding this interaction is necessary to make the most out of CH 4 mitigation feeding strategies using different dietary carbohydrates. The aim of this study was to test the effect on enteric CH 4 emission, feed intake, and milk production response when cows were fed either grass-clover silage (GCS) or corn silage (CS) as the sole forage source (55% of dry matter, DM), in combination with either barley (BAR) or dried beet pulp (DBP) as a concentrate (21.5% of DM). A total of 24 (half first-parity and half second-parity) cows were used in a crossover design with 2 periods of 21 d each, receiving 2 of 4 diets obtained from a 2 × 2 factorial arrangement of the experimental diet. Feed intake, CH 4 emission metrics, and milk production were recorded at the end of the experimental periods. The diets had NDF concentrations between 258 and 340 g/kg of DM and starch concentrations between 340 and 7.45 g/kg of DM (CS-BAR and GCS-DBP, respectively). The effects of silage and concentrate on dry matter intake (DMI) were additive, with the highest feed intake in cows fed CS-BAR, followed by cows fed CS-DBP, GCS-BAR, and GCS-DBP (21.2, 19.9, 19.1, and 18.3 kg/d, respectively). Energy corrected milk (ECM) yield was not affected by silage source in first parity cows, but it was higher for cows fed CS than cows fed GCS in second parity. The effects of silage and concentrate on CH 4 production (g/d), yield (g/kg of DMI), and intensity (g/kg of ECM) were not additive, as cows fed GCS had similar responses regardless of the concentrate used, but cows fed CS had lower CH 4 production, yield, and intensity when fed BAR instead of DBP. The lower CH 4 production, yield, and intensity in cows fed CS-BAR compared with other diets could be partially explained by the nonlinear relationship between ruminal VFA and carbohydrates (NDF and starch) concentration reported in the literature; however, we observed a linear relationship between the acetate/propionate ratio and CH 4 yield, suggesting possible other effects. The effects of silage and concentrate on the ruminal VFA were additive in first parity cows, but not in second parity cows. The interaction between dietary carbohydrate type and parity might indicate an effect of feed intake or the energy balance of the cow. Feeding cows silage and concentrate both rich in starch can result in the lowest enteric CH 4 emission., (The Authors. Published by Elsevier Inc. on behalf of the American Dairy Science Association®. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).)

We determined the effects of increasing the dietary inclusion of whole cottonseed (WCS) on nutrient digestibility and the milk production responses of high-producing dairy cows. Twenty-four multiparous Holstein cows (mean ± SD; 52.7 ± 2.63 kg/d milk; 104 ± 23 DIM) were randomly assigned to treatment sequences in a replicated 4 × 4 Latin square design with 21-d periods. Treatments were increasing doses of WCS at 0%, 8%, 16%, and 24% DM, with WCS replacing soybean meal and hulls to maintain similar diet nutrient composition (% DM) of NDF (32%), forage NDF (21%), starch (27%), and CP (17%). Total fatty acid (FA) content of each treatment was 1.70%, 2.96%, 4.20%, and 5.40% DM, respectively. Three preplanned contrasts were used to test the linear, quadratic, and cubic effects of increasing dietary WCS. Increasing dietary WCS from 0% to 24% DM quadratically influenced the intake of DM and NDF, with the highest value being the inclusion of 8% WCS, and intakes of 16- and 18-carbon, and total FA, with maximum values obtained up to 24% WCS. Increasing dietary WCS affected digestibility of DM (cubic) and NDF (quadratic), with the lowest values being the inclusion of 8% WCS. Increasing WCS increased 16-carbon digestibility (quadratic) but decreased digestibility of 18-carbon and total FA (both quadratic), with highest and lowest values for the inclusion of 24% WCS, respectively. Increasing dietary WCS quadratically increased absorbed 16- and 18-carbon, and total FA, with maximum values obtained for 24% WCS. Increasing dietary WCS quadratically increased yields of milk, milk fat, milk protein, milk lactose, 3.5% FCM, and ECM, and linearly increased BW gain. The source of milk FA was affected by dietary WCS, with a quadratic decrease in the yield of de novo and mixed milk FA and a quadratic increase in preformed milk FA. Increasing dietary WCS linearly increased trans-10 C18:1 milk FA content. As dietary WCS increased, plasma insulin linearly decreased, and plasma gossypol levels linearly increased. Despite the decrease in total FA digestibility, increasing dietary WCS from 0% to 24% DM increased FA absorption. Increasing the dietary inclusion of WCS up to 16% DM increased milk production responses and DM intake. Under the current dietary conditions, high-producing dairy cows benefited best from a diet containing an inclusion of 8% to 16% WCS in DM., (The Authors. Published by Elsevier Inc. on behalf of the American Dairy Science Association®. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).)

The objectives of this study were to determine the farm-level hyperketolactia (HKL) prevalence, as diagnosed from the milk BHB concentration, on dairy farms milking with an automated milking system (AMS) and to describe the farm-level housing, management, and nutritional risk factors associated with increased farm-average milk BHB and the within-herd HKL prevalence in the first 45 DIM. Canadian AMS farms (n = 162; eastern Canada, n = 8; Quebec, n = 24; Ontario, n = 75; western Canada n = 55) were visited once between April and September 2019 to record housing and herd management practices. The first test milk data for each cow under 45 DIM were collected, along with the final test of the previous lactations for all multiparous cows, from April 1, 2019, to September 30, 2020. The first test milk BHB was then used to classify each individual cow as having or not having HKL (milk BHB ≥0.15 mmol/L) at the time of testing. Milk fat and protein content, milk BHB, and HKL prevalence were summarized by farm and lactation group (all, primiparous, and multiparous). During this same time period, formulated diets for dry and lactating cows, including ingredients and nutrient composition, and AMS milking data were collected. Data from the AMS were used to determine milking behaviors and milk production of each herd during the first 45 DIM. Multivariable regression models were used to associate herd-level housing, feeding management practices, and formulated nutrient composition with first test milk BHB concentrations and within-herd HKL levels separately for primiparous and multiparous cows. The within-herd HKL prevalence for all cows was 21.8%, with primiparous cows having a lower mean prevalence (12.2 ± 9.2%) than multiparous cows (26.6 ± 11.3%). Milk BHB concentration (0.095 ± 0.018 mmol/L) and HKL prevalence for primiparous cows were positively associated with formulated prepartum DMI and forage content of the dry cow diet; however, they were negatively associated with formulated postpartum DMI, the major ingredient in the concentrate supplemented through the AMS, and the partially mixed ration to AMS concentrate ratio. Multiparous cows' milk BHB concentration (0.12 ± 0.023 mmol/L) and HKL prevalence were positively associated with the length of the previous lactation, milk BHB at dry-off, prepartum diet nonfiber carbohydrate content, and the major forage fed on farm, while tending to be negatively associated with feed bunk space during lactation. This study is the first to determine the farm-level risk factors associated with herd-level prevalence of HKL in AMS dairy herds. The results may help to optimize management and guide diet formulation and thus promote the reduction of HKL prevalence., (The Authors. Published by Elsevier Inc. on behalf of the American Dairy Science Association®. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).)

The objective of this study was to evaluate the effect of energy and protein in the diet on the recovery of milk ethanol stability (MES) induced by feed restriction. Twelve Holstein and Holstein × Jersey crossbred cows with an average of 146 ± 50 DIM, 575.4 ± 70 kg of BW, and 18.93 ± 5.46 kg/d of milk yield were distributed in a 3 × 3 Latin square design with 3 treatments and 3 experimental periods. Each experimental period lasted 24 d, comprising 3 phases: a 13-d adaptation phase (100E+100P), a 4-d induction phase for milk ethanol instability (50E+50P), and a 7-d recovery phase for MES (3 treatments). The 3 treatments during the recovery phase consisted of 3 diets aiming to meet the requirements of energy and protein (100E+100P), only energy (100E+50P), or only protein (50E+100P). The diet during the adaptation and induction phases was common for all cows. The energy and protein levels to meet each cow's requirements were based on the group average. Restriction of energy and protein reduced DM, CP, and total digestive nutrient intake for cows fed 100E+50P and 50E+100P. The lowest BW was observed for cows fed 50E+100P, with no difference for BCS. During the induction phase, MES was reduced by 9 percentage units. Cows fed 100E+100P recovered MES in the first days of the recovery phase, whereas 100E+50P slightly improved MES, and 50E+100P had a constant decrease in MES. Cows fed 100E+50P and 50E+100P produced, respectively, 3.6 and 5.9 kg less milk than those fed 100E+100P. The 50E+100P treatment exhibited the highest milk fat content and SCS, along with the lowest milk lactose content. Protein content was higher in the 100E+100P treatment. Cows fed 50E+100P showed higher serum albumin levels compared with those on the 100E+100P treatment, not differing from the 100E+50P treatment. We concluded that the complete recovery of MES in cows with feed restrictions is possible only by supplying both the energy and protein requirements in the cows' diet. However, restricting energy intake poses a greater limitation on MES recovery compared with restricting protein., (The Authors. Published by Elsevier Inc. on behalf of the American Dairy Science Association®. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).)

Mare milk has a unique protein composition that makes it a preferred option for adult and infant nutrition. Several functional properties have been attributed to this milk, but with little current evidence. In fact, knowledge on mare milk composition is still limited. In particular, studies addressing the performance of mare milk proteins during human gastrointestinal digestion are scarce, which limits the understanding of mare milk nutritional quality and functionality. For this reason, the present study describes the digestibility of mare milk proteins and the release of peptides as affected by management and lactation stage, factors which are known to affect milk composition. Mare milk samples from 3 different farms collected during 6 mo of lactation (n = 54) were subjected to a static in vitro gastrointestinal model to measure peptide release and protein digestibility. In the present study, a detailed description of protein and individual amino acid behavior during the digestion process was given. For the first time, digestion of the 2 equine β-lactoglobulin isoforms (I and II) was described individually. In addition, it was found that lactation stage and management system can significantly affect protein digestibility and peptide release during gastrointestinal digestion of mare milk. Presumably, differences in the composition of mare milk influence the protein structure and enzyme accessibility, which might have an impact on digestion behavior. Although no specific bioactive peptides were identified, several precursors of previously described bioactive peptides were found. These findings could support the idea of mare milk as a food with added value., (The Authors. Published by Elsevier Inc. on behalf of the American Dairy Science Association®. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).)

The purpose of this experiment was to determine if nicotinic acid (NA) effects on dairy cows and rumen microbial characteristics are forage-type dependent (corn silage, CS; grass silage, GS). Four late-lactation (DIM = 225 ± 12 d) Holstein cows were used in a 4 × 4 Latin square design with a 2 × 2 factorial arrangement of treatments. The main effects were a CS (66.10% CS) based diet or a GS (79.59%) based diet with or without 12 g/d NA. Each experimental period lasted for 28 d. Milk production and milk components, blood metabolites, apparent total-tract nutrient digestibilities, minutes rumen pH were below 5.8 as an indicator of ruminal acidosis, and body temperature changes were analyzed as indicators of heat stress. Nicotinic acid supplementation did not improve apparent total-tract nutrient digestibility. Feeding a GS-based diet improved NDF and hemicellulose digestibility. Feeding a CS-based diet increased the apparent total-tract digestibility of fat, and minutes rumen pH below 5.8 for a greater proportion of the time. The CS-based diet also improved milk yield, milk fat and protein yields, and ECM yield; however, somatic cell count and BHB were also increased. Supplementing NA tended to decrease nonesterified fatty acids, especially when combined with GS where DMI was low. There was a trend for the total protozoa population to increase when GS and NA were fed but decreased when CS and NA were fed. Nicotinic acid tended to decrease rumen protozoal populations of Dasytricha, but increased populations of Ophryoscolex and Diplodiniinae with GS diets and decreased with CS diets. Entodiniinae were increased with CS but NA had no effect. Body temperature was increased when a CS-based diet was fed when compared with a GS-based diet. More research is needed to determine how NA can affect rumen microbial protein synthesis and what kind of diets will provide the optimum effect., (The Authors. Published by Elsevier Inc. on behalf of the American Dairy Science Association®. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).)

The objectives of this observational cohort study were to evaluate the associations of rumination time (RT) in the last week of pregnancy with transition cow metabolism, inflammation, health, and subsequent milk production, reproduction, and culling. Pregnant nulliparous (n = 199) and parous (n = 337) cows were enrolled 21 d before expected calving. Rumination time and physical activity were monitored automatically by sensors from d -21 to 15 relative to calving. Blood samples were collected on d -14, -5, 4, 8, and 12 ± 1 relative to calving. Diagnoses of clinical health problems were performed by researchers from calving to 15 DIM. In classification 1, cows were ranked based on average daily RT in the last week of pregnancy and classified into terciles as short RT (SRT), moderate RT (MRT), or long RT (LRT) for association analyses. In classification 2, RT deviation from the parity average was used in a receiver operating characteristic curve to identify the best threshold to predict postpartum clinical disease. Cows were then classified as above the threshold (AT) or below the threshold (BT). Compared with cows with LRT, cows with SRT had greater serum concentrations of nonesterified fatty acids (0.47 vs. 0.40 ± 0.01 mmol/L), BHB (0.58 vs. 0.52 ± 0.01 mmol/L), and haptoglobin (0.22 vs. 0.18 ± 0.008 g/L) throughout the transition period, and reduced concentrations of glucose, cholesterol, albumin, and magnesium in a time-dependent manner. Parous cows with SRT had higher odds of postpartum clinical disease (adjusted odds ratio [AOR] = 3.7; 95% CI: 2.1-6.4), lower odds of pregnancy by 210 DIM (AOR = 0.34; 95% CI = 0.15-0.75), and lower milk production (46.9 vs. 48.6 ± 0.5 kg/d) than parous cows with LRT. Deviation in prepartum RT had good predictive value for clinical disease in parous cows (area under the curve = 0.65; 95% CI = 0.60-0.71) but not in nulliparous (area under the curve = 0.51; 95% CI = 0.42-0.59). Separation of parous cows according to the identified threshold (≤-53 min from the parity average) resulted in differences in serum concentrations of nonesterified fatty acids (AT = 0.31 ± 0.006, BT = 0.38 ± 0.014 mmol/L), BHB (AT = 0.49 ± 0.008, BT = 0.53 ± 0.015 mmol/L), and globulin (AT = 32.3 ± 0.3, BT = 34.8 ± 0.5 g/L) throughout the transition period, as well as in serum cholesterol, urea, magnesium, albumin, and haptoglobin in a time-dependent manner. Below threshold parous cows had higher odds of clinical disease (AOR = 3.7; 95% CI = 2.1-6.4), reduced hazard of pregnancy (adjusted hazard ratio = 0.64, 95% CI: 0.47-0.89), greater hazard of culling (adjusted hazard ratio = 2.1, 95% CI: 1.2-3.6), and lower milk production (46.3 ± 0.7 vs. 48.5 ± 0.3 kg/d). External validation using data from 153 parous cows from a different herd and the established threshold in RT deviation (≤-53 min) resulted in similar predictive value, with the odds of postpartum disease 2.4 times greater in BT than AT (37.5% vs. 20.1%). In conclusion, RT in the week preceding calving was a reasonable predictor of postpartum health and future milk production, reproduction, and culling in parous cows but not in nulliparous cows., (The Authors. Published by Elsevier Inc. on behalf of the American Dairy Science Association®. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).)

Most trace minerals (TM) are fed above dairy cow requirements in commercial herds but their fate and effects on dairy cows have not been well documented. In this study, we evaluated the effects of feeding short-term sulfate TM above recommendations on apparent total-tract digestibility of nutrients, rumen fermentation characteristics, serum concentrations, and milk yield and composition, as well as milk, fecal, and urinary TM excretion in midlactation dairy cows. Eight multiparous Holstein cows with an average body weight (± SD) of 684 ± 29 kg at 82 ± 10 DIM in a quadruple 2 × 2 crossover design were fed a basal diet, differing in sulfate TM supplement concentrations, to provide either 0.11, 17, and 63 (control; CON) or 0.95, 114, and 123 (high trace minerals; HTM) mg of dietary Co, Mn, and Zn per kilogram of DM, respectively. Each experimental period had a 21-d adaptation to the diet, followed by a 10-d sample collection period. Feed ingredients and total feces and urine were collected during 4 consecutive d and rumen fluid was collected 0, 1, 2, 4, and 6 h relative to feeding. Milk yield was recorded daily, and milk samples were collected on 4 consecutive milkings. Ingestion of Co, Mn, and Zn was higher for the HTM group compared with the CON group by 216%, 233%, and 93%, respectively. Dry matter intake averaged 25.0 (SE = 0.6) kg/d, and apparent total-tract digestibility of major nutrients was similar between treatments. High trace minerals had no measurable effect on ruminal pH, major volatile fatty acids, and protozoa counts. Isovalerate molar proportion was 9.4% greater for the HTM group compared with the CON group. Neither milk yield (43.5 kg/d; SE = 0.8) nor milk fat and protein concentrations differed between treatments. Milk urea nitrogen concentration was significantly higher for HTM (11.7 mg/dL) compared with CON (9.7 mg/dL; SE = 0.7). Fecal excretion of Co, Mn, and Zn increased by 223%, 198%, and 75%, respectively, for the HTM group compared with the CON group. Urinary excretions of TM were marginal compared with feces, and only urinary Co and Mn were significantly higher for HTM cows than CON cows, as was similarly obtained for serum Co and Mn concentrations. Milk TM yields were not modified by treatments. In summary, short-term dietary sulfate TM supply over the recommendation did not improve cow performance but significantly increased fecal TM excretion, which could affect TM accumulation in soils where manure is applied and could potentially result in leaching into nearby watersheds. Further studies are needed to evaluate the impact of high fecal TM excretion on the environment using the One Health approach. Moreover, the effects of TM oversupply on milk production and cow health should be evaluated by long-term experiments., (Copyright © 2024 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

Omitting or shortening the dry period may result in a fairly constant ration throughout the transition period of dairy cows, reducing the need for adaptation of cow metabolism and rumen function to a new lactation. The objective of this study was to determine the effect of dry period length (DP) on rumen adaptation and cow metabolic state during the transition period. Twelve pregnant, rumen-cannulated Holstein Friesian dairy cows at the end of their first lactation were assigned to 1 of 3 treatments: a conventional (60 d), short (30 d), or no dry period (0 d). At dry-off, cows received a dry-cow ration until calving. Lactating cows received a lactation ration. Cows were monitored from 8 wk before calving until 8 wk after calving for milk yield and DMI. Rumen biopsies were taken from 3 locations in the rumen at 60, 40, and 10 d before calving and 3, 7, 14, 28, and 56 d after calving to assess papillae dimensions. Blood was sampled weekly from 3 wk before until 8 wk after calving, and liver biopsies were taken at wk -2, 2, and 4 relative to calving. Prepartum, DMI, and milk yield were greater for cows with a short or no dry period, compared with cows with a conventional dry period. Postpartum, DMI was greater for cows with a short dry period compared with cows with a conventional dry period. Plasma glucose concentration was greater for cows without a dry period, compared with the other DP postpartum. Plasma concentrations of nonesterified fatty acids and BHB, and liver triglyceride content, did not differ among dry period. Rumen papillae differed in size based on biopsy location, but there was no interaction between biopsy location and the effect of DP. Rumen papillae surface area for cows managed for a 30- or 60-d dry period decreased toward calving. At 40 d prepartum, papillae surface area was greater for short and no dry period treatment compared with a conventional dry period. At 10 d prepartum, papillae surface area was greater for the no dry period treatment compared with both other treatments, and this difference was still present 3 d postpartum. Cows managed for a short dry period showed faster increase in papillae dimensions after calving compared with cows managed for a conventional dry period. From d 28 onwards, no differences in papillae surface area were observed. The faster rumen adaptation postpartum may be related to the increased DMI during the first weeks postpartum for cows managed for a short dry period. However, this did not result in improved metabolic status or milk yield. The results from the present study demonstrate that the dietary changes related to a conventional DP affected rumen papillae development, not only prepartum but also early postpartum. Further optimization of DP as well as dietary composition throughout the transition period may support cows in their adaptation to a new lactation., (The Authors. Published by Elsevier Inc. on behalf of the American Dairy Science Association®. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).)

Animals vary in the way in which they use energy due to diet, genetics, and management. Energy consumed by the animal supports milk production, but considerable variation among animals in energy utilization is thought to exist. The study objective was to estimate the among-animal variance in energy utilization in data collected from Jersey cows using indirect calorimetry. Individual animal-period data from 15 studies (n = 560) were used. The dataset included 115 animals from 44 to 410 DIM producing 11.5 to 39.1 kg/d of milk. On average, the 63 treatments in the dataset ranged 14.8% to 19.5% CP, 21.4% to 43.0% NDF, 16.2% to 33.3% starch, and 2.21% to 6.44% crude fat. Data were analyzed with the GLIMMIX procedure of SAS (9.4) with random effects of cow, treatment nested within period, square, and experiment. The percentage of among-animal, dietary treatment, and experimental variance was calculated as the variance associated with each fraction divided by the sum of variance from animal, dietary treatment, experiment, and residual, which was considered the total variance. The percentage of among-animal variance was characterized as high or low when the value was greater than or less than the mean value of 29.2%. Among-animal variance explained approximately 29.3% to 42.5% of the total variance in DMI, gross energy (GE), digestible energy (DE), ME, and NE L in megacalories per day. When energetic components of feces, urine, heat, and lactation in megacalories per day were expressed per unit of DMI, the among-animal variance decreased by 20.4, 4.82, 9.55, and 3.80 percentage units, respectively. However, among-animal variance explained 4.80 and 5.02 percentage units more of the total variation for methane energy and tissue energy in megacalories per day when expressed per unit of DMI. Variance in energetic efficiencies of DE/GE, ME/GE, and ME/DE were explained to a lesser extent by among-animal variance (averaging 17.8 ± 1.95%). The among-animal contribution to total variance in milk energy was 28.8%. Milk energy was a large proportion of the energy efficiency calculation, which included milk energy plus corrected tissue energy over net energy intake, which likely contributed to the 22.2% of total among-animal variance in energy efficiency. Results indicate that among-animal variance explains a large proportion of the total variation in DMI. This contributes to the variance observed for energy fractions as well as energy components when expressed in megacalories per day. Variation in energetic loss associated with methane was primarily explained by differences among animals and was increased when expressed per unit of DMI, highlighting the role of inherent animal differences in these losses., (The Authors. Published by Elsevier Inc. on behalf of the American Dairy Science Association®. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).)

Improving the low productivity levels of native cattle breeds in smallholder farming systems is a pressing concern in Pakistan. Crossbreeding high milk-yielding holstein friesian (HF) breed with the adaptability and heat tolerance of Sahiwal cattle has resulted in offspring that are well-suited to local conditions and exhibit improved milk yield. The exploration of how desirable traits in crossbred dairy cattle are selected has not yet been investigated. This study aims to provide the first overview of the selective pressures on the genome of crossbred dairy cattle in Pakistan. A total of eighty-one crossbred, thirty-two HF and twenty-four Sahiwal cattle were genotyped, and additional SNP genotype data for HF and Sahiwal were collected from a public database to equate the sample size in each group. Within-breed selection signatures in crossbreds were investigated using the integrated haplotype score. Crossbreds were also compared to each of their parental breeds to discover between-population signatures of selection using two approaches: cross-population extended haplotype homozygosity and fixation index. We identified several overlapping genes associated with production, immunity, and adaptation traits, including U6, TMEM41B, B4GALT7, 5S&#95;rRNA, RBM27, POU4F3, NSD1, PRELID1, RGS14, SLC34A1, TMED9, B4GALT7, OR2AK3, OR2T16, OR2T60, OR2L3, and CTNNA1. Our results suggest that regions responsible for milk traits have generally experienced stronger selective pressure than others., (© 2024. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.)

Recent research has highlighted the importance of dietary fatty acid profile of fatty acid supplements on production responses of high-producing dairy cows. Conventional soybeans contain ∼15% oleic acid and ∼50% linoleic acid whereas high oleic acid soybeans (HOSB) contain ∼70% oleic acid and ∼7% linoleic acid. We determined the effect of increasing dietary inclusion of roasted and ground HOSB on production responses of high-producing dairy cows. Twenty-four multiparous Holstein cows (mean ± SD; 50.7 ± 4.45 kg/d of milk; 122 ± 57 DIM) were randomly assigned to treatment sequences in a replicated 4 × 4 Latin square design with 21-d periods. Treatments were increasing doses of HOSB at 0%, 8%, 16%, and 24% of DM. The HOSB replaced conventional soybean meal and hulls to maintain similar diet nutrient composition (% of DM) of 27.4% to 29.4% NDF, 20.6% forage NDF, 27.5% starch, and 15.9 to 16.5% CP. Total fatty acid content of treatments was 1.65%, 3.11%, 4.52%, and 5.97% of DM, respectively. Preplanned polynomial orthogonal contrasts included the linear, quadratic, and cubic effects of increasing HOSB. Increasing dietary inclusion of HOSB linearly decreased DMI and MUN and increased yields of milk, 3.5% FCM, ECM, and milk fat, and quadratically increased milk protein. The increased response to milk fat was due to an increase in preformed milk fatty acids. Due to the increase in milk component yields and decrease in DMI, there was an increase in feed efficiency. Increasing HOSB inclusion linearly decreased plasma BUN concentration and tended to decrease plasma insulin. Increasing HOSB had no effect on BW change or BCS change. In summary, increasing dietary inclusion of HOSB up to 24% of DM increased production responses of high-producing dairy cows and did not affect body reserves., (The Authors. Published by Elsevier Inc. on behalf of the American Dairy Science Association®. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).)

Feed efficiency is important for economic profitability of dairy farms; however, recording daily DMI is expensive. Our objective was to investigate the potential use of milk mid-infrared (MIR) spectral data to predict proxy phenotypes for DMI based on different cross-validation schemes. We were specifically interested in comparisons between a model that included only MIR data (model M1); a model that incorporated different energy sink predictors, such as body weight, body weight change, and milk energy (model M2); and an extended model that incorporated both energy sinks and MIR data (model M3). Models M2 and M3 also included various cow-level variables (stage of lactation, age at calving, parity) such that any improvement in model performance from M2 to M3, whether through a smaller root mean squared error (RMSE) or a greater squared predictive correlation (R 2 ), could indicate a potential benefit of MIR to predict residual feed intake. The data used in our study originated from a multi-institutional project on the genetics of feed efficiency in US Holsteins. Analyses were conducted on 2 different trait definitions based on different period lengths: averaged across weeks versus averaged across 28 d. Specifically, there were 19,942 weekly records on 1,812 cows across 46 experiments or cohorts and 3,724 28-d records on 1,700 cows across 43 different cohorts. The cross-validation analyses involved 3 different k-fold schemes. First, a 10-fold cow-independent cross-validation was conducted whereby all records from any one cow were kept together in either training or test sets. Similarly, a 10-fold experiment-independent cross-validation kept entire experiments together, whereas a 4-fold herd-independent cross-validation kept entire herds together in either training or test sets. Based on cow-independent cross-validation for both weekly and 28-d DMI, adding MIR predictors to energy sinks (model M3 vs. M2) significantly (P < 10 -10 ) reduced average RMSE to 1.59 kg and increased average R 2 to 0.89. However, adding MIR to energy sinks (M3) to predict DMI either within an experiment-independent or herd-independent cross-validation scheme seemed to demonstrate no merit (P > 0.05) compared with an energy sink model (M2) for either R 2 or RMSE (respectively, 0.68 and 2.55 kg for M2 in herd-independent scheme). We further noted that with broader cross-validation schemes (i.e., from cow-independent to experiment-independent to herd-independent schemes), the mean and slope bias increased. Given that proxy DMI phenotypes for cows would need to be almost entirely generated in herds having no DMI or training data of their own, herd-independent cross-validation assessments of predictive performance should be emphasized. Hence, more research on predictive algorithms suitable for broader cross-validation schemes and a more earnest effort on calibration of spectrophotometers against each other should be considered., (The Authors. Published by Elsevier Inc. on behalf of the American Dairy Science Association®. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).)

The aim of this study was to evaluate two levels of forage allowance (FA) during the prepartum period on metabolic, hormonal, productive, and reproductive variables in primiparous (P) and multiparous (M) beef cows. Six weeks before calving, 40 P and 42 M cows were assigned to two FA (native pastures) treatments until calving (day 0): High (15 kg DM/100 BW; H) and Low (5 kg DM/100 BW; L). After calving all cows were managed together. High FA cows presented greater intake than L cows, while M tended to have greater intake than P cows. Increased herbage allowance in late gestation in beef cows successfully improved forage intake and energy balance reflected in body condition score and metabolic and endocrine markers. Also, high forage allowance increased milk yield (5.4 vs 4.6 kg/d, P < 0.05) and calves daily gain rate during the first two months of age (0.88 vs 0.82 kg/d, P < 0.05) in H respect to L cows. Pregnancy rate was greater in H vs L cows (40 vs 28 %, P < 0,05). The endocrine metabolic response to higher forage allowance depended on parity, as primiparous cows showed a greater imbalance around calving. Moreover, most primiparous cows were not pregnant at the end of the breeding season, consistent with their prolonged anestrous. Multiparous cows had better productive and reproductive performance than primiparous cows, showing that growing cows have a pronounced negative energy balance reflected in most of the evaluated parameters., Competing Interests: Declaration of competing interest The authors declare no conflict of interest. The funders had no role in the design of the study; in the collection, analyses, or interpretation of data; in the writing of the manuscript; or in the decision to publish the results., (Copyright © 2024 Elsevier Inc. All rights reserved.)

At the onset of lactation in dairy cows, inflammation and oxidative stress may occur and result in a risk of pathologies and lower milk yield. To propose an innovative management strategy for cows during this period, it is essential to better understand these physiological variations. Our objective was to evaluate the metabolic, redox, and immune status of 7 primiparous and 8 multiparous Holstein cows during late gestation and the first months of lactation. Blood samples were collected between 3 wk before calving until 12 wk postpartum. Milk samples were also collected, but only at the time points after calving. The metabolic (nonesterified fatty acids [NEFA], BHB, glucose, urea, calcium) and redox (reactive oxygen metabolites [ROM], oxidative stress index [OSI], glutathione peroxidase activity, vitamin E) statuses were analyzed in plasma or erythrocytes. The expression of genes related to antioxidant functions was determined in leukocytes collected from milk. For immune status, plasma cytokine levels and the production of reactive oxygen species (ROS) in classical and regulatory neutrophils were measured in 2 whole blood ex vivo challenges. The data were analyzed using a mixed model that included the fixed effects of parity and week and their interaction. Milk yield, plasma NEFA, and BHB in wk 2 and 4 after calving were higher in multiparous cows than in primiparous cows, whereas glucose and calcium tended to be lower. Plasma ROM and OSI levels in wk 8 were higher in multiparous than in primiparous cows. Multiparous cows also displayed higher glutathione peroxidase activity in erythrocytes, and antioxidant transcription factor and superoxide dismutase-1 expression levels in milk leukocytes. Moreover, multiparous cows had higher plasma concentrations of vitamin E but lower plasma levels of cytokines CXCL10, CCL2, IL1Rα, and IFNγ. Following ex vivo whole blood stimulation with Escherichia coli, lower IL1α and TNFα levels were measured in multiparous than in primiparous cows. Intracellular ROS production by neutrophils was lower in multiparous than in primiparous cows. These results thus indicated marked physiological changes in wk 8 compared with wk 2 and 4 of lactation. These differences in the physiological status of primiparous and multiparous cows offer interesting perspectives for potential dietary strategies to prevent pathologies which take account of parity and week relative to calving., (The Authors. Published by Elsevier Inc. on behalf of the American Dairy Science Association®. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).)

Nutrition and physiological state affect hepatic metabolism. Our objective was to determine whether feeding flaxseed oil (∼50% C18:3n-3 cis), high oleic soybean oil (∼70% C18:1 cis-9), or milk fat (∼50% C16:0) alters hepatic expression of PC, PCK1, and PCK2 and the flow of carbons from propionate and pyruvate into the tricarboxylic acid (TCA) cycle in preruminating calves. Male Holstein calves (n = 40) were assigned to a diet of skim milk with either: 3% milk fat (MF; n = 8), 3% flaxseed oil (Flax; n = 8), 3% high oleic soybean oil (HOSO; n = 8), 1.5% MF + 1.5% high oleic soybean oil (MF-HOSO; n = 8), or 1.5% MF + 1.5% flaxseed oil (MF-Flax; n = 8) from d 14 to d 21 postnatal. At d 21 postnatal, a liver biopsy was taken for gene expression and metabolic flux analysis. Liver explants were incubated in [U- 13 C] propionate and [U- 13 C] pyruvate to trace carbon flux through TCA cycle intermediates or with [U- 14 C] lactate, [1- 14 C] palmitic acid, or [2- 14 C] propionate to quantify substrate oxidation to CO 2 and acid-soluble products. Compared with other treatments, plasma C18:3n-3 cis was 10 times higher and C18:1 cis-9 was 3 times lower in both Flax (Flax and MF-Flax) treatments. PC, PCK1, and PCK2 expression and flux of [U- 13 C] pyruvate as well as [U- 13 C] propionate were not different among treatments. PC expression was negatively correlated with the enrichment of citrate M+5 and malate M+3, and PCK2 was negatively correlated with citrate M+5, suggesting that when expression of these enzymes is increased, carbon from pyruvate enters the TCA cycle via PC-mediated carboxylation, and then oxaloacetate is converted to phosphoenolpyruvate via PCK2. Acid-soluble product formation and PC expression were reduced in HOSO (MF-HOSO and HOSO) treatments compared with Flax (MF-Flax and Flax), indicating that fatty acids regulate PC expression and carbon flux, but that fatty acid flux control points are not connected to PC, PCK1, or PCK2. In conclusion, fatty acids regulate hepatic expression of PC, PCK1, and PCK2, and carbon flux, but the point of control is distinct., (The Authors. Published by Elsevier Inc. on behalf of the American Dairy Science Association®. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).)

Milk production and overall dairy farm economics depend on rearing dairy heifers. This study investigated the presence of a genotype by environment interaction in Holstein (HOL), Nordic Red dairy cattle (RDC), and their F 1 crossbreeds (HOL × RDC) when provided different feed rations. The aim of our study was to assess how different energy concentrations in feed rations affect growth, BCS, feed intake, and feed efficiency in the 3 groups during the prepubertal period. The 3 breed groups were randomly allocated to receive either a standard or a low-energy feed ration. Holstein heifers exhibited reduced growth and a lower BCS when they were fed the low-energy feed ration. In contrast, the RDC heifers demonstrated similar growth rates with the different feed rations and maintained similar BCS irrespective of feed energy concentration. The HOL × RDC crossbred heifers performed as an intermediate between the HOL and RDC groups. Significant differences were observed in DMI and energy intake in the HOL and HOL × RDC groups depending on feed ration treatment. The RDC heifers had similar feed intake irrespective of treatment. There were no significant differences in the feed conversion ratio among breeds and feed treatments. These results indicate the presence of a genotype by environment interaction in prepubertal HOL and RDC heifers in response to differences in feed ration treatment. Due to the influence of prepubertal growth on future milk production, reproduction, and health status, it is important to be aware of breed-specific requirements during the prepubertal period, particularly in mixed breed and crossbred groups, to optimize growth rates and production potential., (The Authors. Published by Elsevier Inc. on behalf of the American Dairy Science Association®. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).)

The effects of high (2.5% of DM) versus normal dietary sodium chloride (NaCl) intake on renal urea-N kinetics and urea-N metabolism were investigated in 9 rumen-cannulated and multicatheterized lactating dairy cows in a crossover design with 21-d periods. It was hypothesized that urinary urea-N excretion would be greater, and BUN concentration lower in response to greater diuresis induced by high NaCl intake. Also, urea-N transport across ruminal and portal-drained viscera (PDV) tissues was hypothesized to be affected by dietary sodium intake. A second experiment was conducted using 8 lactating cows in a crossover design with 14-d periods to test high NaCl (2.5% of DM) versus high KCl (3.2% of DM) intake on milk yield and MUN concentrations. Experiment 1 showed that despite greater diuresis, high NaCl intake had no effect on urinary urea-N excretion or BUN concentration. The high NaCl intake did not affect rumen ammonia concentrations, total rumen VFA concentrations, ruminal venous - arterial concentration differences for ammonia, or ammonia absorption indicating that high NaCl did not adversely affect ruminal fermentation and microbial protein synthesis. High NaCl intake did not affect the total amount of urea-N transport from blood to gut, but ruminal venous - arterial concentration differences for urea-N were lower with high NaCl and ruminal extraction of arterial urea-N was numerically smaller, indicating that the ruminal epithelial urea-N transport was lower with high NaCl. Energy-corrected milk yield was greater with high NaCl (3.2 ± 1.5 kg/d); however, MUN concentrations were not affected by treatment. In experiment 2, ECM was greater with NaCl (1.4 ± 0.31 kg/d) compared with KCl (30.2 and 28.8 ± 0.91 kg ECM/d, respectively). Milk urea-N concentration was lower with KCl, suggesting a urea-N lowering effect in milk not evident with high NaCl intake. In conclusion, the present data show that dietary Na intake of 12 to 13 g/kg DM was followed by greater diuresis but did not affect urea-N excretion or BUN concentration. High NaCl intake did not affect the total amount of urea-N transfer across PDV tissues. Energy-corrected milk yield was greater with high NaCl compared with both control and feeding KCl; however, with KCl, MUN decreased., (The Authors. Published by Elsevier Inc. on behalf of the American Dairy Science Association®. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).)

Background: Gamma-aminobutyric acid (GABA) is an important neurotransmitter in the human body, with several negative emotions reported as being associated with GABA dysregulation. This study investigates the safety and modulatory effects of GABA-enriched milk, fermented by Streptococcus thermophilus GA8 and Lacticasebacillus rhamnosus HAO9, on the gut microbiota and neurotransmitter profiles in mice., Results: Through rigorous culturing and fermentation processes, we achieved consistent GABA production in milk, with concentrations reaching 4.6 and 8.5 g L -1 for GA8-fermented and co-fermented milk, respectively, after 48 h. Using SPF male C57BL/6J mice, we administered either mono-culture or combined-culture milk treatments and monitored physiological impacts. The treatments did not affect mouse body weight but induced significant changes in gut microbiota composition. Beta diversity analysis revealed distinct microbial profiles between treatment groups, highlighting fermentation-specific microbial shifts, such as an increase in Verrucomicrobia for the GA8 group and a modulation in Saccharibacteria&#95;genera&#95;incertae&#95;sedis for the GA8 + HAO9 group. Serum neurotransmitter levels were elevated in both treatment groups, with significant increases in l-glutamine, l-tryptophan and, notably, serotonin hydrochloride in the GA8 + HAO9 group. Correlation analysis identified a positive association between specific bacterial genera and neurotransmitter levels, suggesting a probiotic effect on neuroactive substances., Conclusion: These findings suggest that fermented milk has potential as a probiotic supplement for mood improvement and stress relief, highlighting its role in modulating the gut-brain axis. © 2024 Society of Chemical Industry., (© 2024 Society of Chemical Industry.)

Supplemental dietary rumen available fats show promise as enteric methane (eCH 4 ) mitigators for lactating dairy cows. However, concerns include variability in eCH 4 response and possible negative effects on dairy cow performance. Successful implementation of this mitigation option requires better prediction of responses specifically to rumen available fatty acids (FA) as well as understanding the modulating effects of other dietary and animal characteristics. Using meta-analytic and meta-regression techniques, 35 published studies with diet definition were used to assess changes in eCH 4 emissions and lactation performance associated with supplemental fat, specific supplemental rumen available FA types, and other dietary characteristics. Enteric CH 4 (g/d) was reduced by 3.77% per percentage unit of supplemental rumen available ether extract (EE). Supplemental rumen available PUFA (C18:2 and C18:3) and UFA (C18:1, C18:2, C18:3) mitigated eCH 4 (g/d) emissions in dairy cows by 6.88 and 4.65% per percentage unit increase, respectively. The anti-methanogenic effects of PUFA, MUFA and MCFA increased with correspondingly greater basal dietary levels of each FA type. Higher rumen-degradable starch (RDS; >18% DM) in the basal diet promoted greater reductions in eCH 4 yield (eCH 4 /DMI, g/kg) with supplemental rumen available PUFA and UFA. Both milk fat percentage and yield (kg/d) were reduced with rumen available fat supplementation with a reduction of 7.8% and 6.0%, respectively, relative to control diets. Our results highlight the importance of determining basal levels of the rumen available FA before providing supplemental rumen available FA as an option for enteric eCH 4 mitigation. Dairy nutritionists can use estimates generated from this analysis to predict changes in eCH 4 emissions and dairy cow performance associated with dietary supplementation of rumen available EE and specific rumen available FA types for the purpose of eCH 4 mitigation., (The Authors. Published by Elsevier Inc. on behalf of the American Dairy Science Association®. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).)

Pistachio hull (PH), a rich source of tannin, may interact with nitrogen (N) and affect N metabolism in dairy cows. This investigation aimed to assess the effects of feeding PH and two N sources (soybean meal: SBM or slow-release urea: SRU) on milk production, digestibility, and N metabolism in lactating dairy cows. The study utilized a 4 × 4 Latin square design with 12 Holstein dairy cows (body weight = 611 ± 39 kg; 84 ± 14 days in milk; 45.6 ± 5.3 kg/d milk), with treatments arranged in a 2 × 2 factorial configuration. The main factors were (1) the tannin source [without or with dried PH included at 76.5 g/kg of diet DM] and (2) two N sources [SBM vs. SRU]. Crude protein and total phenolic and tannin contents were greater, but NDF was lower in PH than in sugar beet pulp. Feeding diets containing 7.65% PH resulted in reductions in milk yield, milk urea N, and milk efficiency but milk fat and protein concentration increased. Milk yield was similar between SBM and SRU. Feeding PH in replacement to sugar beet pulp did not interact with N sources, except for feed intake, as the PH + SRU diet tended to decrease feed intake (P = 0.09). Feeding PH was also associated with decreased dry matter, crude protein, and NDF digestibility. Dry matter digestibility decreased when a portion of SBM was replaced by SRU. Feeding PH in replacement to sugar beet pulp lowered ruminal ammonia-N concentration and increased acetate-to-propionate proportion. Feeding PH vs. sugar beet pulp had no effect on N intake and N excretion in urine and milk, but it increased the amount of N excretion in feces and decreased apparent N efficiency (milk N/N intake). Alanine aminotransferase was lower in cows fed SRU only in PH- but not sugar beet pulp-containing diets. Blood creatinine was lower when SRU was included in the PH-containing diet, but the opposite was observed in diets containing sugar beet pulp. Feeding PH instead of sugar beet pulp did not affect blood urea-N but resulted in lower glucose and aspartate aminotransferase concentrations. Overall, cows fed PH in replacement to sugar beet pulp were less efficient in converting feed nutrients into milk production, likely because of its negative effect on nutrient digestion. Partial replacement of SBM with SRU had no influence on lactation productivity and urea-N concentration in milk and blood. This may imply the suitability of SRU in providing a sustained supply of N in the rumen, likely qualifying it as a viable source of N to replace a portion of SBM in dairy diets., (© 2024. The Author(s), under exclusive licence to Springer Nature B.V.)

Aim : We hypothesized that milk, yogurt, and cheese have differential impacts on energy expenditure (EE) and obesity in mice fed a high-fat diet (HFD). Methods : C57BL/6 mice ( n = 16 per group) were fed a HFD or a HFD supplemented with fat-free milk (MILK), fat-free plain yogurt (YOG), or reduced-fat cheddar cheese (CHE; 19 kcal% fat), each provided at 10% of the daily energy intake, for 8 weeks. EE was quantified using a metabolic chamber. Metabolic pathways related to BAT mitochondrial function and uncoupling protein 1 (UCP1) abundance were assessed. Serum lipidomic profiles were analyzed to identify potential mediators of the observed effects. Results : MILK supplementation lowered weight gain and fat accumulation and enhanced EE and BAT thermogenesis, perhaps via the SIRT1-AMPK-PGC1α axis in BAT. This led to elevated UCP1 abundance and enhanced the abundance of hormone-sensitive lipase (HSL). MILK also altered serum lipid species, indicating enhanced energy use, and promoted BAT thermogenesis and mitochondrial function pathways. YOG exhibited a similar pattern but a lower magnitude of effects than MILK on reducing weight gain and fat mass, increasing EE, and BAT thermogenic proteins, including AMPK-PGC1α-UCP1. Both MILK and YOG showed a relative increase in serum PC 15:0&#95;15:0 and LPC 15:0. In contrast, CHE reduced weight gain and increased EE without impacting BAT thermogenesis proteins or serum lipid species. Conclusion : Our study showed that MILK, YOG, and CHE reduced weight gain in mice on a HFD by increasing EE. MILK and YOG also up-regulated BAT thermogenesis, while both additionally altered lipids involved in fat metabolism and inflammation. CHE did not affect BAT thermogenesis and lipid species compared to HFD.

This study aimed to estimate genetic parameters, including heritability and repeatability, for milk yield and prolificacy in sheep. It included 3682 records from 1837 ewes across various breeds: Awassi, Assaf, and Awassi x Assaf crosses, two Awassi lines: Improved Awassi, AFEC Awassi. The study measured total milk yield (TMY), yield up to 120 days (TMY120), and yield up to 150 days (TMY150), alongside reproductive traits: litter size (LS), number of lambs born alive (NLBA), and lambing interval (LI). The analysis utilized a mixed model and the REML procedure to estimate genetic parameters. Results indicated that litter size (LS) had no significant impact on milk traits, whereas breed, location, ewe parity, and lambing season showed significant effects. Lactation length also significantly influenced TMY. For reproductive traits, treatment was significant for NLBA, with location associated with breed, parity, and season affecting all traits. Heritability estimates for TMY ranged from 0.00 in Awassi x Assaf crosses to 0.11 in Awassi, and for TMY120 and TMY150, from 0.00 in Awassi x Assaf crosses to 0.16 in Awassi. Estimates for LS and NLBA varied similarly. The heritability for LI was 0.03 in Awassi and zero in other breeds and crosses. The findings suggest that in the northern West Bank regions of Nablus and Jenin, Assaf or Awassi x Assaf breeds are preferable. The study underscores the importance of comprehensive performance and pedigree recording for effective sheep farm management., (© 2024. The Author(s), under exclusive licence to Springer Nature B.V.)

Fat supplements are well known for their multiple beneficial effects on ruminant health, reproduction and productivity, and as a source for certain bioactive compounds in ruminant products. On the other hand, numerous phytochemicals have demonstrated the potential to improve rumen fermentation through modifying the volatile fatty acid (VFA) pattern to favour those with greater energy efficiency, boosting microbial protein synthesis, and decreasing methane emission and ruminal ammonia concentration. Savory is an aromatic plant rich in various phytochemicals (mainly carvacrol and flavonoids) that can alter ruminal metabolism of dietary fatty acids, potentially increasing the production of some bioactive compounds such as conjugated linoleic acids (CLAs). This study aimed to investigate combined effects of oil type (fish oil (FO) versus soybean oil (SO)) and the inclusion of savory (Satureja khuzistanica) plant (SP) in the diet on total tract digestibility of nutrients, rumen fermentation, milk yield and milk fatty acid profile in dairy cattle. Eight multiparous lactating Holstein cows were used in a replicated 4 × 4 Latin square design experiment with four diets and four 21-d periods. During each experimental period consisted of 14 days of adaptation and a 7-day sampling period, cows were randomly assigned to one of the four dietary treatments: the diet supplemented with 2% (DM basis) fish oil (FOD) or soybean oil (SOD), the FOD or SOD plus 370 g DM/d/head SP (FODs, SODs, respectively). The experimental diets were arranged in a 2 × 2 factorial design, with the fat sources as the first and SP as the second factor. The FO-supplemented diets had lower dry matter intake (DMI) and higher total tract digestibility than SO-supplemented diets (P < 0.05), and including SP in the diet improved total tract digestibility of dry matter (DM), organic matter (OM), ether extract (EE), and non-fibrous carbohydrates (NFC) (P < 0.05) without negatively affecting DMI. Rumen pH was lower with SO than with FO diets (P < 0.01) and increased with SP inclusion in the diet (P < 0.05). Total protozoa count and ruminal ammonia concentration decreased, and the branched-chain VFA (BCVFA) proportion increased with SP inclusion in the diet (P < 0.05). Milk production, as well as the concentration and yield of milk components (except lactose concentration) were higher with SO than with FO diets (P < 0.05), but these variables remained unaffected by SP. The milk concentrations of both non-esterified fatty acids (NEFA) and beta-hydroxybutyrate (BHB) were lower with SO compared to FO diets, and these variables were reduced by SP (P < 0.01). The proportions of both mono- and polyunsaturated FA (MUFA and PUFA, respectively) in milk were higher with FO than with SO diets (P < 0.01), and their proportions increased by SP at the expense of saturated FA (SFA) (P < 0.01). Including SP in the diet increased the proportions of all the milk n-3 FA (C18:3c, C20:5, and C22:6) by 21%, 40%, and 97%, respectively, and those of conjugated linoleic acids (C18:2 (c9,t11-CLA) and C18:2 (t10,c12-CLA)) by 23% and 62%, respectively. There was no interaction between oil type and SP for the assessed variables. Fish oil, despite reducing milk production and milk components, was more effective than soybean oil in enriching milk with healthy FA. These findings also show promise for SP as a feed additive with the potential to improve total tract digestibility, rumen fermentation and milk FA composition., (© 2024. The Author(s).)

Precise as well as unbiased estimates of genetic parameters of economically important traits is important for framing breeding programmes meant for genetic progress of dairy animals. Present work was carried out to assess the co-variances along with genetic parameters of production and reproduction traits in Sahiwal cattle with Bayesian approach. Records of Sahiwal cattle kept at ICAR-National Dairy Research Institute, Karnal, Haryana for the period 1988-2016 were used. Heritability estimates ± posterior standard deviation for 305 days' milk yield (305DMY), lactation milk yield (LMY), lactation length (LL), peak yield (PY), days to attain peak yield (DPY), dry period (DP), gestation period (GP), calving to first insemination interval (CFI), days open (DO) and calving interval (CI) as obtained were 0.23 ± 0.03, 0.26 ± 0.03, 0.18 ± 0.03, 0.18 ± 0.03, 0.07 ± 0.02, 0.08 ± 0.02, 0.08 ± 0.02, 0.07 ± 0.02, 0.09 ± 0.02 and 0.02 ± 0.02, respectively and estimates of repeatability for the corresponding traits were 0.23 ± 0.03, 0.26 ± 0.03, 0.21 ± 0.03, 0.21 ± 0.03, 0.12 ± 0.02, 0.14 ± 0.03, 0.16 ± 0.03, 0.16 ± 0.03, 0.17 ± 0.03 and 0.17 ± 0.03. Moderate heritability values for 305DMY and LMY implies that sufficient additive genetic variability is available to obtain selection response. Repeatability estimates for 305DMY, LMY and PY were moderate, pointing towards the possibility of early appraisal of Sahiwal cattle for selection. Antagonistic genetic associations were observed between reproduction and production traits suggesting that reproduction traits need due attention while selecting high producing Sahiwal animals., (© 2024. The Author(s), under exclusive licence to Springer Nature B.V.)

The aim of this study was to assess the impact of genotype-environment interaction (GEI) on the manifestation of traits such as age at first calving (AFC), age at first service (AFS), and calving interval (CI) through the application of the reaction norm model in Holstein cattle raised in Paraná state, Brazil. Utilizing data from the milk testing service of the Paraná Association of Holstein Cattle Breeders (APCBRH), this study analyzed records from 179,492 animals undergoing their first, second, and third lactations from the years 2012 to 2022. These animals were part of 513 herds spread across 72 municipalities in Paraná. The environmental gradient was established by normalizing contemporary group solutions, derived from the animal model, with the 305-day-corrected milk yield serving as the dependent variable. Subsequently, reaction norms were determined utilizing a Random Regression Model. Spearman's correlation was then applied to compare the estimates of breeding values across different environmental gradients for the studied traits. The highest EG (+ 4) indicates the least challenging environments, where animals experience better environmental conditions. Conversely, lower EG (-4) values represent the most challenging environments, where animals endure worse conditions. The only trait that exhibited a moderate heritability magnitude was AFC (0.23) in the least challenging environmental condition. The other traits were classified as having low heritability magnitudes regardless of the evaluated environmental gradient. While minimal evidence was found for the influence of GEI on CI, a clear GEI effect was observed for AFC and AFS across all environmental gradients examined. A reversal in genotype ranking occurred under extreme environmental conditions. The findings suggest that the best-performing genotype under one environmental gradient may not necessarily excel under another., (© 2024. The Author(s), under exclusive licence to Springer Nature B.V.)

Cosmetically applicable soluble agonists for Toll-like receptor 2 (TLR2), which can strengthen skin barrier function, were produced by fermentation of asparagus ( Asparagus officinalis L.) extract supplemented with skimmed milk using Lactobacillus delbrueckii subsp. lactis TL24. Their molecular size was estimated to be >100 kDa. Their TLR2-stimulating activity was stable over 1 year at 4 °C, but it decreased by more than 95% within 10 and 4 months at 25 °C and 40 °C, respectively. The possibility of stabilization of TLR2-stimulating activity by powdering was tested, and we found that lyophilization with 10% or a higher amount of dextrin could stabilize the activity even at 40 °C. The powdered fermented product dose-dependently stimulated TLR2. It augmented the formation of tight junctions in normal human keratinocytes, as detected by fluorescence staining of occludin and ZO-1, whereas their protein and gene expression levels did not increase, suggesting that a change in subcellular localization of these proteins without significant changes in their amounts might be responsible. The powder nature has some benefits over the aqueous, besides stability, e.g., it can be dissolved just before application, allowing fresh material to be used each time, and it may widen a range of cosmetic applications in non-aqueous types of cosmetics.