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Dietary fat content can reduce the methane production of dairy cows; however, the relevance fatty acid (FA) composition has towards this inhibitory effect is debatable. Furthermore, in-depth studies elucidating the effects of unsaturated fatty acids (UFA) on rumen function and the mechanism of reducing methane (CH4) production are lacking. This study exposed 10 Holstein cows with the same parity, similar milk yield to two total mixed rations: low unsaturated FA (LUFA) and high unsaturated FA (HUFA) with similar fat content. The LUFA group mainly added fat powder (C16:0 > 90%), and the HUFA group mainly replaced fat powder with extruded flaxseed. The experiment lasted 26 d, the last 5 d of which, gas exchange in respiratory chambers was conducted to measure gas emissions. We found that an increase in the UFA in diet did not affect milk production (P > 0.05) and could align the profile of milk FAs more closely with modern human nutritional requirements. Furthermore, we found that increasing the UFA content in the diet lead to a decrease in the abundance of Methanobrevibacter in the rumen (|linear discriminant analysis [LDA] score| > 2 and P < 0.05), which resulted in a decrease in the relative abundance of multiple enzymes (EC:1.2.7.12, EC:2.3.1.101, EC:3.5.4.27, EC:1.5.98.1, EC:1.5.98.2, EC:6.2.1.1, EC:2.1.1.86 and EC:2.8.4.1) during methanogenesis (P < 0.05). Compared with the LUFA group, the pathway of CH4 metabolism was inhibited in the HUFA group (|LDA| > 2 and P < 0.05), which ultimately decreased CH4 production (P < 0.05). Our results illustrated the mechanism involving decreased CH4 production when fed a UFA diet in dairy cows. We believe that our study provides new evidence to explore CH4 emission reduction measures for dairy cows.

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After birth, the immune system is challenged by numerous elements of the extrauterine environment, reflected in fluctuations of inflammatory markers. The concentrations of these markers in the first month of life are associated with the future performance of dairy youngstock. It is thought that bacterial genera colonizing the calf intestinal tract can cause inflammation and thus affect their host's performance via immunomodulation. This study explored how the faecal microbiota of newborn dairy calves were related to inflammatory markers during the first three weeks of life, and if the abundance of specific genera was associated with first-lactation performance. Ninety-five female Holstein calves were studied. Once a week, serum and faecal samples were collected, serum concentrations of serum amyloid A, haptoglobin, tumour necrosis factor-α, and interleukin-6 were measured, and faecal microbiota composition was examined by 16S rRNA gene amplicon sequencing. Faecal Gallibacterium abundance in the first week of age and Collinsella abundance in the second week were negatively associated with inflammatory response as well as with calving-conception interval. Peptostreptococcus abundance in the second week of life was positively associated with inflammatory response and calving-conception interval, and negatively with average daily weight gain. In the third week, Dorea abundance was positively, Bilophila abundance was negatively associated with inflammatory response, and both genera were negatively associated with age at first calving. These bacterial genera may be able to influence the inflammatory response and through this, possibly the future performance of the dairy heifer. Deciphering such microbiota-host interactions can help improve calf management to benefit production and welfare.

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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.

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Objective: The aim of this investigation was to examine the impact of Taraxacum coreanum (known as dandelion) (TC) and TC mixtures with milk thistle (MT) or Aspergillus oryzae (AO) as feed additives on the immune response, milk quality, and milk production in Holstein cows over 6 weeks of administration. Materials and Methods: Thirty-two healthy Holstein dairy cows were provided 30 kg of total mixed ration (TMR) with no TC, 90 gm TC, 54 gm TC + 36 gm MT, or 54 gm TC + 36 gm AO 40% groups. The feed additives were supplied daily in two equal portions (per 45 gm) by topdressing the TMR for 6 weeks. Milk and blood samples were collected weekly. Results: In the TC-treated cows (TC, TC + MT, and TC + AO groups), significantly lower peripheral blood white blood cell (WBC) counts at 6 weeks and milk somatic cell counts (SCCs) at 4-6 weeks of administration were observed. Concentrations of serum superoxide dismutase (SOD) and glutathione peroxidase (GSH-px) were notably elevated in cows treated with TC for 4-6 weeks, while levels of proinflammatory cytokines concentrations of tumor necrosis factor-alpha (TNF-α) and chemokine (IL-8) were significantly reduced in TC-treated cows after 3-6 weeks of administration. Conclusion: These results suggested that TC or a TC mixture with other medicinal herbs supplementations enhanced the serum antioxidative activities and, consequently, might suppress the adverse immune response due to lower serum TNF-α and IL-8 release supported by lower WBC and SCC counts.

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Inorganic sources of Mg are commonly used in dairy cow diets, but their availability varies significantly. This study assessed the relative availability of 4 commonly used inorganic Mg sources and a novel alkalinizing proprietary mineral blend [PMB; Multesium (GLC Minerals, LLC, Green Bay, WI, USA)]. The study was a duplicated 6 × 6 Latin square, with 12 nonlactating, non-pregnant Holstein dairy cows assigned to a square based on BW and parity. Cows were fed 90% of their voluntary DMI (diet contained 0.21% Mg). Each experimental period lasted 7 d. On d 2 of each period, urinary catheters were fitted. Total urine collection started on d 3 for 48 h with samples collected and measured every 12 h. On d 4, 30 g of Mg were administered as boluses with gelatin capsules: negative control (one empty capsule), magnesium oxide (MgO), magnesium sulfate (MgSO4), calcium magnesium hydroxide [CaMg(OH)4], calcium magnesium carbonate [CaMg(CO3)2], and PMB [a blend of Ca and Mg sources that includes CaMg(CO3)2, CaMg(OH)4, and MgO]. Blood samples were collected at 0, 1, 2, 3, 12, and 24 h after treatment administration on d 4 of each treatment period. Urine and blood samples were analyzed for Mg and Ca concentration. Statistical analyses were conducted with PROC GLIMMIX including treatment, time, period, square, treatment × time, treatment × period, and time × period as fixed effects, and cow nested within square as a random effect in the model. Urinary Mg excretion for 4 of the Mg sources studied [PMB, MgO, CaMg(OH)4, and MgSO4] increased significantly, representing an increase of at least 40.8% relative to control. The supplementation of CaMg(CO3)2 did not significantly increase relative to control. There were no significant changes in blood Mg concentration with treatment; but, a significant treatment × time effect was observed. Calcium-rich sources [PMB, CaMg(OH)4, CaMg(CO3)2] had lower blood Mg concentrations at 12 or 24 h after treatment than control and CaMg(CO3)2. Based on urinary Mg excretion 24 h after treatment, 4 of the Mg sources evaluated (including PMB) showed a similar availability, however, the availability of the commercial CaMg(CO3)2 source included in our study was similar to the negative control (no-supplemented cows).

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Pregnancy is a complex process involving complex molecular interaction networks, such as between miRNA-protein, protein-protein, metabolite-metabolite, and protein-metabolite interactions. Advances in technology have led to the identification of many pregnancy-associated microRNA (miRNA), protein, and metabolite fingerprints in dairy cows. An array of miRNA, protein, and metabolite fingerprints produced during the early pregnancy of dairy cows were described. We have found the in silico interaction networks between miRNA-protein, protein-protein, metabolite-metabolite, and protein-metabolite. We have manually constructed miRNA-protein-metabolite interaction networks such as bta-miR-423-3p-IGFBP2-PGF2α interactomes. This interactome is obtained by manually combining the interaction network formed between bta-miR-423-3p-IGFBP2 and the interaction network between IGFBP2-PGF2α with IGFBP2 as a common interactor with bta-miR-423-3p and PGF2α with the provided sources of evidence. The interaction between bta-miR-423-3p and IGFBP2 has many sources of evidence including a high miRanda score of 169, minimum free energy (MFE) score of -25.14, binding probability (p) of 1, and energy of -25.5. The interaction between IGFBP2 and PGF2α occurs at high confidence scores (≥0.7 or 70%). Interestingly, PGF2α is also found to interact with different metabolites, such as PGF2α-PGD2, PGF2α-thromboxane B2, PGF2α-PGE2, and PGF2α-6-keto-PGF1α at high confidence scores (≥0.7 or 70%). Furthermore, the interactions between C3-PGE2, C3-PGD2, PGE2-PGD2, PGD2-thromboxane B2, PGE2-thromboxane B2, 6-keto-PGF1α-thromboxane B2, and PGE2-6-keto-PGF1α were also obtained at high confidence scores (≥0.7 or 70%). Therefore, we propose that miRNA-protein-metabolite interactomes involving miRNA, protein, and metabolite fingerprints of early pregnancy of dairy cows such as bta-miR-423-3p, IGFBP2, PGF2α, PGD2, C3, PGE2, 6-keto-PGF1 alpha, and thromboxane B2 may form the key regulatory networks and players of pregnancy regulation in dairy cows. This is the first study involving miRNA-protein-metabolite interactomes obtained in the early pregnancy stage of dairy cows.

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The objectives of this study were to evaluate mucosal immune responses in peripartum Holstein cows, to assess the impact of intranasal modified live viral (MLV) vaccination on mucosal immunity, and to explore the relationship between genotype and peripartum immune responses. Eighty multiparous Holstein cows were randomized to receive either: 1) intranasal MLV tri-valent viral vaccine 18-24 days prior to expected calving (DC); 2) the same vaccine within twelve hours after parturition (F); 3) vaccine at both time points (DCF), or 4) no vaccine (CON). Nasal secretions and sera were collected from all cattle pre-vaccination and on multiple days before and after calving to determine concentrations of interferon beta (IFN-beta) and IFN-gamma and bovine herpesvirus-1 (BHV-1-) and bovine respiratory syncytial virus (BRSV-) specific IgA in nasal secretions, and BHV-1 and BRSV serum neutralizing (SN) titers. Cows were genotyped by bead-based microarray, genotypes were used to categorize previously established health traits, and relationships between immune responses and genotype were evaluated. There was no significant effect of vaccination on immune responses, although all vaccinated groups demonstrated numerically increased IFN-gamma within four days post vaccination. There was a significant (P <0.0001) time effect on nasal IgA in CON, F, and DCF groups, with the highest nasal IgA titers measured post calving. There was a significant (P <0.0001) time effect on nasal IFN-beta in all groups. Significant relationships between genotype and immune response were not detected. Contrary to previous reports of systemic immunosuppression, bovine mucosal responses appear to be intact in the peripartum period.

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A new video based multi behavior dataset for cows, CBVD-5, is introduced in this paper. The dataset includes five cow behaviors: standing, lying down, foraging,rumination and drinking. The dataset comprises 107 cows from the entire barn, maintaining an 80% stocking density. Monitoring occurred over 96 h for these 20-month-old cows, considering varying light conditions and nighttime data to ensure standardization and inclusivity.The dataset consists of ranch monitoring footage collected by seven cameras, including 687 video segment samples and 206,100 image samples, covering five daily behaviors of cows. The data collection process entailed the deployment of cameras, hard drives, software, and servers for storage. Data annotation was conducted using the VIA web tool, leveraging the video expertise of pertinent professionals. The annotation coordinates and category labels of each individual cow in the image, as well as the generated configuration file, are also saved in the dataset. With this dataset,we propose a slowfast cow multi behavior recognition model based on video sequences as the baseline evaluation model. The experimental results show that the model can effectively learn corresponding category labels from the behavior type data of the dataset, with an error rate of 21.28% on the test set. In addition to cow behavior recognition, the dataset can also be used for cow target detection, and so on.The CBVD-5 dataset significantly influences dairy cow behavior recognition, advancing research, enriching data resources, standardizing datasets, enhancing dairy cow health and welfare monitoring, and fostering agricultural intelligence development. Additionally, it serves educational and training needs, supporting research and practical applications in related fields. The dataset will be made freely available to researchers world-wide.

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Short-chain fatty acids (SCFAs) produced in the rumen are key factors affecting dairy cows' energy balance (EB). This study aimed to quantitatively evaluate the effects of SCFAs production on EB in dairy cows. Primiparous dairy cows were divided into high non-esterified fatty acid (NEFA; group H) and low NEFA (group L) groups based on their blood NEFA levels at week 3 postpartum, which served as an indicator of EB. The amounts of SCFAs produced in the rumen, including acetate, propionate, and butyrate (SCFAsP), were calculated using the predicted rumen volume. Because there were no differences between the groups in SCFAsP/dry matter intake, whereas 4% fat-corrected milk (FCM)/SCFAsP was significantly higher in group H, it was suggested that more body fat was mobilized for milk production in group H. However, group L, which showed better EB, had propionate dominant and lower FCM/SCFAsP and milk energy/SCFAs energy at 3 and 7 weeks postpartum, indicating that group L had a better energy supply for milk production. These results suggest that SCFAsP produced by rumen fermentation and the composition of SCFAs in the rumen affect milk production and EB.

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Today, one of the major challenges of dairy farmers is to reduce their environmental footprint to establish more effective, efficient, and sustainable production systems. Feed additives such as yeast probiotics could potentially allow them to achieve these objectives through the improvement of milk production, feed efficiency, and ration valorization, hence mitigating the environmental impacts of milk production. In this study, the life cycle assessment (LCA) principle was performed to estimate the environmental impact of the production and supplementation of a commercial yeast probiotic (Actisaf Sc 47) in three trials performed in three different countries that are representative for around 50% of the milk production in Europe: France (French trial), United Kingdom (UK trial), and Germany (German trial). For each trial, two groups of animals were compared: control, without Actisaf Sc 47 supplementation, used as baseline; and experimental, with Actisaf Sc 47 supplementation at 5 or 10 g/cow/day. Different impact categories were analyzed for each group to calculate the impact of producing 1 kg of fat- and protein-corrected milk. An initial analysis was done only during the period of Actisaf Sc 47 supplementation and showed than the supplementation with Actisaf Sc 47 reduced, on average by 5%, the carbon footprint during the three trials. A second analysis was done via the extrapolation of all the data of each trial to an annual farm level, including the lactation period (305 days), dry period (60 days), and the period with and without Actisaf Sc 47 supplementation. Reported at a farm annual scale, the average reduction allowed by Actisaf Sc 47 supplementation was 2.9, 2.05, 2.47, 1.67, 2.28, 2.18, 2.14, and 2.28% of the carbon footprint, land use, water use, resource use, acidification, freshwater eutrophication, marine eutrophication, and terrestrial eutrophication, respectively. On average, the production of 1 kg of fat- and protein-corrected milk by using Actisaf Sc 47 was shown to improve environmental impacts compared to control. Regarding Actisaf Sc 47 production, the LCA showed that the production of 1 kg of Actisaf Sc 47 emitted 2.1 kg CO2 eq with a negligible contribution to total the carbon footprint of milk ranging from 0.005 to 0.016%. The use of Actisaf Sc 47 in dairy cows could then result in different positive outcomes: improving performance and efficiency while reducing the global carbon footprint.

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Artificial insemination 12 h following observed standing estrus is a standard estimate of the fertility levels of cattle. The main objective of this study was to determine if controlling ovarian development with a fertility program could alter the fertility of lactating dairy cows. Lactating dairy cows (n = 1356) 60-66 days in milk (DIM) were randomly assigned to receive timed AI following Presynch-11/Ovsynch (Presynch-11) or a combination of estrus detection and timed AI using the Presynch-11/Ovsynch program (Estrus + TAI). Cows in standing estrus, following the first two cloprostenol sodium (CLO) injections, in the Estrus + TAI group were artificially inseminated using the AM/PM rule. Cows in this group that were not observed in standing estrus received Ovsynch and TAI beginning 11 d after the second CLO injection. Cows in the Presynch-11 group received two injections of 500 µg CLO 14 d apart but were not observed for estrus. The first GnRH (100 µg) of Ovsynch was administered 11 d following the second CLO injection. All cows in the Presynch-11-Ovsynch group received TAI following Ovsynch. Cows in this treatment were then assigned randomly to receive either CLO or dinoprost 7 d following the first GnRH of Ovsynch. The final GnRH of Ovsynch was administered 56 h later and TAI 16 h after the final GnRH. Pregnancies/AI (P/AI) were greater (p < 0.001) for the Presynch-11 group compared with the Estrus + TAI group (45 vs. 31%). Primiparous cows had greater fertility following observed standing estrus compared with multiparous cows. Days in milk at the first AI were greater (p < 0.01) in the Presynch-11 group vs. the Estrus + TAI group (98 vs. 80) but less variable (p < 0.01). The range of DIM at the first AI was 95 to 101 in the Presynch-11 group and 60 to 101 in the Estrus + TAI group. Within the Presynch-11 group, there were no differences in the rate of luteolysis or P/AI for the prostaglandin type at the final PGF2α of Ovsynch. Multiparous cows treated with Presynch-11 had >60% chance of pregnancy compared with multiparous cows receiving AI following standing estrus. In summary, lactating dairy cows receiving timed AI following the Presynch-11/Ovsynch program had improved fertility compared with a group of cows that received AI following standing estrus or, if not observed in estrus, timed AI. This comparison indicated that controlling ovarian development with GnRH and PGF2α positively impacted the fertility of lactating dairy cows.

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This study describes the association of non-esterified fatty acids (NEFA) and calcium concentrations at calving with early lactation disease, reproductive performance and culling in 646 dairy cows from 13 commercial grazing dairy herds in Uruguay. During one year, health events were recorded from calving to 30 days in milk (DIM). The first author visited each farm every 20 days. During each visit, body condition score (BCS) was recorded (scale 1-5), defining BCS < 3 as suboptimal and BCS > 3 as optimal, and a blood sample was taken from cows between 0 and 4 DIM for metabolite determination. To evaluate the association between health events (i.e., retained placenta-metritis and clinical mastitis) and risk factors (parity, BCS, high NEFA (> 0.6 mmol/L) and subclinical hypocalcemia (SCH) (< 2.10 mM)) data were analysed using multivariable logistic regression models. To evaluate the association of health events and risk factors with reproductive performance and culling, data were analysed using Cox proportional hazard regression models. A risk factor and an outcome of interest were assumed to be associated at P < 0.05 and a tendency to be associated was defined at P < 0.10. Overall, 47 % (n = 303) of the cows showed elevated NEFA concentration and 77 % (n = 499) had SCH. In addition, 21.5 % (n = 139) of the cows recorded at least one clinical disease. Cumulative incidence was 17 % (n = 109) for clinical mastitis, 4.2 % (n = 27) for retained placenta (RP)-metritis and 1.4 % (n = 7) for lameness. Clinical mastitis was associated with parity, with lower odds in primiparous (PP) cows (OR = 0.42, P < 0.01). Cows in an optimal BCS also tended to have lower odds (OR = 0.66, P = 0.07). Moreover, high NEFA and SCH cows had higher odds of CM (OR = 4.5, P = 0.01 and OR = 1.75, P = 0.04, respectively). Retained placenta-metritis tended to be associated with high NEFA concentration (OR = 2.2, P = 0.06). Primiparous cows with suboptimal BCS showed an increased first insemination rate (HR = 2.34; P < 0.01). The risk of culling was lower in PP cows (HR = 0.19; P < 0.01) and in cows with optimal BCS and low NEFA concentration (HR = 0.38; P = 0.03). Our data show that metabolic challenge (defined as peripartum suboptimal BCS, high NEFA or SCH) is associated with increased odds of clinical mastitis and RP-metritis, decreased probability of insemination and increased hazard of culling. Under grazing conditions, we suggest that farm management to improve the metabolic adaptation to lactation represents an opportunity to enhance cow performance in terms of health, fertility and longevity.

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To boost the immune function around parturition, recombinant bovine granulocyte colony-stimulating factor (rbG-CSF) has been used to increase the number of neutrophils. Therefore, the aim of this study was to quantify the effect of rbG-CSF administration on the incidence of postpartum pathologies, reproductive performance, and milk production during the first three months of lactation. A total of 199 Holstein cows from one herd were included and were randomly allocated into two groups: Control (n = 103) and rbG-CSF (n = 96). Cows in the rbG-CSF group received 2 doses of a rbG-CSF injectable formulation, one 7 days before the expected date of calving and the other within 24 h after calving. For 6 weeks following calving, animals were examined weekly to assess the presence of postpartum pathologies. Milk production, protein and fat content, and somatic cell count were determined monthly by the regional dairy herd improvement association. Data about the reproductive performance were collected from on-farm software. To analyse the effect of treatment on the incidence of postpartum pathologies, Pearson's χ2 test and multivariable logistic regressions were performed. The effect on reproductive performance was analysed using Cox proportional hazard regression analysis for days open, binary logistic regression for first service conception rate and Oneway ANOVA test for the number of artificial inseminations. The effects of treatment on milk yield and milk composition were checked using GLM repeated measures analysis. No statistically significant differences were observed between treatment groups for any of the parameters evaluated. Only parity had a significant effect on days open and milk production (p < 0.05). In conclusion, in the present study no evidence was found that rbG-CSF could have an effect on the reproductive and productive parameters evaluated.

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Melatonin, an endogenous indolamine derived from tryptophan, is primarily synthesized by the pineal gland in mammals and regulated by a complex neural system. Its release follows a circadian rhythm, which is crucial for regulating physiological processes in response to light-dark cycles in both humans and animals. In this review, we report that the presence of this hormone in bovine milk, with significant differences in concentration between daytime and nighttime milking, has increased interest in milk as a natural source of bioactive molecules. Melatonin lowers cortisol levels at night, reduces body temperature and blood pressure, coinciding with decreased alertness and performance, acts as an antioxidant and anti-inflammatory agent, modulates the immune system, offers neuroprotective benefits, and supports gastrointestinal health by scavenging free radicals and reducing oxidative stress in dairy cows. Many factors influence the release of melatonin, such as the intensity of artificial lighting during nighttime milking, the frequency of milkings, milk yield, and genetic differences between animals. Nocturnal milking under low-intensity light boosts melatonin, potentially reducing oxidative damage and mastitis risk. Additionally, ultra-high temperature (UHT) treatment does not significantly affect the melatonin content in milk. However, further research on its stability during milk processing and storage is crucial for ensuring product efficacy. In some countries, nighttime milk with naturally elevated melatonin content is already commercialized as a natural aid for sleep. Thus, naturally melatonin-rich milk may be a promising alternative to synthetic supplements for promoting better sleep and overall well-being.

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Our preliminary study identified dairy cow placenta extract (CPE) as a mixture of peptides with potent antioxidant activity both in vivo and in vitro. However, the specific antioxidant peptides (AOPs) responsible for this activity were not yet identified. In the current study, we employed virtual screening and chromatography techniques to isolate two peptides, ANNGKQWAEVF (CP1) and QPGLPGPAG (CP2), from CPE. These peptides were found to be less stable under extreme conditions such as high temperature, strong acid, strong alkali, and simulated digestive conditions. Nevertheless, under normal physiological conditions, both CP1 and CP2 exhibited significant antioxidant properties, including free-radical scavenging, metal chelating, and the inhibition of lipid peroxidation. They also up-regulated the activities of intracellular antioxidant enzymes in response to hydrogen-peroxide-induced oxidative stress, resulting in reduced MDA levels, a decreased expression of the Keap1 gene and protein, and increased levels of the Nrf2 and HO-1 genes and proteins. Furthermore, CP1 demonstrated superior antioxidant activity compared to CP2. These findings suggest that CP1 and CP2 hold potential for mitigating oxidative stress in vitro and highlight the efficacy of virtual screening as a method for isolating AOPs within CPE.

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The metabolites secreted by probiotics or released after their lysis are called postbiotics. They provide physiological benefits to the host, preventing the colonisation of pathogens by improving the intestinal environment for beneficial commensal bacteria, which reduces the incidence of digestive disorders and improves the immune system. The aim of this work was to evaluate the addition of postbiotics to dairy cow rations during the transition period on nutrient digestibility, composition, and milk yield. The effects of two postbiotics were evaluated in twelve Friesian cows from 30 days before calving to two months of lactation. The animals were randomly allocated to two treatment groups: control (CT) and supplemented with postbiotics (PC and PR). Feeding was ad libitum with 60/40 of forage/concentrate ratio on dry matter basis. Daily feed intake and milk production were recorded individually throughout the study. Two digestibility balances were performed, one before parturition and one after parturition. Colostrum was sampled at first milking and milk was sampled weekly. Data were analysed using a mixed mode in R software 4.4.1. The results suggest that postbiotic supplementation in late gestation and early lactation increase the voluntary intake of dry matter, especially in the PR treatment, with higher apparent total tract digestibility of dry matter, organic matter and neutral detergent fibre. Both treatments including postbiotics induced an increase in colostral immunoglobulin concentration. Milk production of cows receiving the PC treatment was the highest, with high fat and protein yields and a higher persistence of the production curve throughout the lactation.

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Insulin-like growth factor 1 (IGF-1) regulates dairy cow reproduction, while the paracrine IGF system locally influences fertility. In both systems, IGF-1 bioactivity is regulated through binding proteins (IGFBPs) inhibiting IGF-1 binding to its receptor (IGF1R). This study aimed to investigate a possible transfer between this endocrine and paracrine system. Therefore, blood and follicular fluid (FF) from postpartum dairy cows were analysed for ß-hydroxybutyrate (BHB), IGF-1, IGFBP-2, -3, -4, -5, and an IGFBP fragment in two study parts. The mRNA expression of IGFBP-2, IGFBP-4, IGF1R, and the pregnancy-associated plasma protein A (PAPP-A) in granulosa cells was measured. The results showed correlations between plasma and FF for IGF-1 (r = 0.57, p < 0.001) and IGFBP-2 (r = -0.57, p < 0.05). Blood BHB negatively correlated with IGF-1 in blood and FF and IGFBP-3, -5 and total IGFBP in blood (IGF-1 plasma: r = -0.26, p < 0.05; FF: r = -0.35, p < 0.05; IGFBP-3: r = -0.64, p = 0.006; IGFBP-5: r = -0.49, p < 0.05; total IGFBP: r = -0.52, p < 0.05). A negative correlation was found between IGFBP-2 expression and IGF-1 concentration in FF (r = -0.97, p = 0.001), while an IGFBP fragment positively correlated with IGF1R-mRNA in FF (r = 0.82, p = 0.042). These findings suggest a transfer and local regulation between the somatotropic axis and the follicular IGF system, linking the metabolic status with local effects on dairy cow fertility.

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We predicted the energy balance of cows from milk traits and estimated the genetic correlations of predicted energy balance (PEB) with fertility traits for the first three lactations. Data included 9,646,606 test-day records of 576,555 Holstein cows in Japan from 2015 to 2019. Genetic parameters were estimated with a multiple-trait model in which the records among lactation stages and parities were treated as separate traits. Fertility traits were conception rate at first insemination (CR), number of inseminations (NI), and days open (DO). Heritability estimates of PEB were 0.28-0.35 (first lactation), 0.15-0.29 (second), and 0.09-0.23 (third). Estimated genetic correlations among lactation stages were 0.85-1.00 (first lactation), 0.73-1.00 (second), and 0.64-1.00 (third). Estimated genetic correlations among parities were 0.82-0.96 (between first and second), 0.97-0.99 (second and third), and 0.69-0.92 (first and third). Estimated genetic correlations of PEB in early lactation with fertility were 0.04 to 0.19 for CR, -0.03 to -0.19 for NI, and -0.01 to -0.24 for DO. Genetic improvement of PEB is possible. Lower PEB in early lactation was associated with worse fertility, suggesting that improving PEB in early lactation may improve reproductive performance.

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Level of dam milk production (DMP) and dam milk fat to protein ratio (DFPR), as an indicator of metabolic status in dairy cows, have been identified to be associated with productive and reproductive performance of the offspring. Yet whether the interaction of DMP by DFPR can be associated with performance of the offspring have not been studied to our knowledge. Therefore, the present study was conducted to investigate the association of the main and interactive effects of DMP and DFPR with offspring's birth weight, survival, milk yield and fertility. To this end, data of birth weight, culling rate, milk yield and reproductive variables of offspring born to lactating dams (n = 14,582) and data associated with DMP and DFPR during 305-day lactation were retrieved. Afterwards, offspring were classified in three categories of DMP, including DMP1 (dams with <10.00 × 103 kg of 305-day milk production), DMP2 (dams with ≥10.00 × 103 kg and <14.00 × 103 kg of 305-day milk production), DMP3 (dams with ≥14.00 × 103 kg of 305-day milk production), and three categories of DFPR, including DFPR1 (offspring born to dams with <1.00 FPR), DFPR2 (offspring born to dams with ≥1.00 and < 1.40 FPR) and DFPR3 (offspring born to dams with ≥1.40 FPR). Statistical analysis revealed no association of the interaction effect of DMP by DFPR with investigated variables in the offspring (P > 0.05). However, the main effect of DMP was positively associated with milk yield, but negatively associated with survival, age at first insemination and conception during nulliparity, and transgenerational improvement in milk yield in the offspring (P < 0.05). Moreover, the main effect of DFPR was positively associated with birth weight, survival and first service conception rate during nulliparity, but negatively associated with metabolic status and reproductive performance during primiparity in the offspring (P < 0.05). In conclusion, the present study did not find any interaction effect of DMP by DFPR on productive and reproductive variables in the offspring. This finding implicates the association of DMP with milk production in the offspring was regardless of DFPR. Moreover, this finding implies the association of DFPR with postpartum metabolic status and reproductive performance in the offspring was regardless of DMP.

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Metabolic disorders pose significant challenges in transition dairy cows. Numerous parameters have been investigated in this context, and apelin has recently emerged as a potential metabolic indicator. Accordingly, this study aimed to assess the associations between this hormone and other metabolic parameters. Twenty-two adult Holstein-Friesian dairy cows, 21 days before their expected calving date, were selected for blood sampling and serum separation at four time points: 21 and 10 days before calving and 10 and 21 days after parturition. Serum concentrations of apelin, leptin, insulin, cortisol, T3, T4, non-esterified fatty acids, glucose, total protein, albumin, globulin, aspartate aminotransferase, alanine transaminase, triglycerides, cholesterol, high, low and very low-density lipoproteins, total, direct and indirect bilirubin were measured in these samples. Surrogate indices for insulin resistance, body condition score, and milk production were also evaluated. Throughout the transition period, a significant increase in apelin levels was observed. Various models were employed to identify associations between apelin and the studied metabolic parameters. Notably, significant correlations between apelin and Leptin, Insulin, Cortisol, T3, T4, NEFA, Cholesterol, LDL, VLDL, Total Protein, Albumin, Globulin, Total Bilirubin, Direct Bilirubin and Indirect Bilirubin were observed, with some being immediate while others developed over time. These findings indicate a mutual influence between apelin and specific metabolic indices. Changes in any component of the metabolic profile at one stage can lead to alterations in apelin levels in subsequent stages. The correlations uncovered between apelin and other components of the metabolic profile in transitioning dairy cows offer valuable insights, contributing to a better understanding of the potential effects of apelin on the studied indicators and vice versa.

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