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Background: Breastfeeding offers significant health benefits, but its practice and success can vary. While research on induced lactation in cisgender women has been documented, there is limited research on lactation induction in transgender women., Case Presentation: A 50-year-old transgender woman undergoing hormone therapy and living with a pregnant partner sought to co-feed using induced lactation. After approval by the hospital ethics committee, a regimen of estradiol, progesterone, and domperidone was initiated, accompanied by nipple stimulation. Lactation was successfully induced and maintained, with milk composition analysis indicating high levels of protein and other key nutrients. This case, the seventh reported, highlights the complexity of lactation induction in transgender women, considering factors such as age, obesity, and insulin resistance. The nutrient profile of the milk suggests its suitability for infant feeding, despite some differences from typical human milk., Conclusions: Induced lactation is feasible in transgender women, expanding the understanding of non-puerperal lactation and its potential in diverse family structures. Further research is warranted to optimize lactation induction protocols in transgender women., (© 2024. The Author(s).)

This systematic review and meta-analysis aimed to estimate the prevalence of pregnancy- and lactation-associated osteoporosis in postpartum women within 1 year of delivery. We searched MEDLINE via PubMed and Igaku Chuo Zasshi for articles published in English or Japanese from the inception of the database to September 2021. Two researchers independently screened and included observational studies reporting the prevalence of pregnancy- and lactation-associated osteoporosis in postpartum women within 1 year of delivery. Of the 3,425 screened records, 8 articles centered on postpartum women were included in the review. Seven studies used dual-energy X-ray absorptiometry for assessing bone mineral density, while one used a quantitative ultrasound method. In the seven studies that used dual-energy X-ray absorptiometry, the parameters used to define osteoporosis were the T-score (two studies), Z-score (three studies), both T- and Z-scores (one study), and young adult mean (one study). Evaluation timeframes included 1 week (three studies), 1-2 months postpartum (three studies), and 1 week to 12 months postpartum (one study). The estimated prevalence of pregnancy- and lactation-associated osteoporosis defined by dual-energy X-ray absorptiometry was as follows: lumbar spine (six studies), 5% (95% confidence interval [CI], 0-13; heterogeneity [I 2 ] = 99%) and femoral neck (three studies), 12% (95% CI, 0-30; I 2 = 99%). Pregnancy and lactation were found to elevate the fracture risk in women, underscoring the necessity for a standardized assessment in diagnosing pregnancy- and lactation-associated osteoporosis. This imperative step aims to enable early detection and treatment of bone mineral loss among postpartum women.

Background: Goat milk is gaining popularity as a superior alternative to bovine milk due to its closer resemblance to human milk. Understanding the molecular processes underlying lactation is crucial for improving milk quality and production in goats. However, the genetic mechanisms governing lactation in goats, particularly in indigenous breeds like the Jakhrana, remain largely unexplored., Results: In this study, we performed a comprehensive transcriptomic analysis of Jakhrana goat mammary glands during early and late lactation stages. We isolated milk somatic cells and conducted RNA sequencing, followed by transcript quantification and mapping against the ARS1.2 Capra hircus reference assembly. Our analysis identified differentially expressed genes (DEGs) and commonly expressed genes (CEGs) across the lactation phases. Early lactation showed enrichment of genes encoding antimicrobial peptides and lubrication proteins, while late lactation exhibited heightened expression of genes encoding major milk proteins. Additionally, DEG analysis revealed upregulation of pivotal genes, such as the ABC transporter gene MRP4, implicated in modulating milk composition and quality., Conclusion: Our findings provide insights into the genetic mechanisms underlying lactation dynamics in the Jakhrana goat. Understanding these mechanisms could help in improving milk production and quality in goats, benefiting both the dairy industry and consumers., (© 2024. The Author(s).)

Lactation women, a highly concerned demographic in society, face health risks that deserve attention. Zinc oxide nanoparticles (ZnO NPs) are widely utilized in food and daily products due to their excellent physicochemical properties, leading to the potential exposure of lactating women to ZnO NPs. Hence, assessing the potential risks associated with ZnO NP exposure during lactation is critical. While studies have confirmed that exposure to ZnO NPs during lactation can induce toxic responses in multiple organs through blood circulation, the effects of lactational exposure on mammary tissue remain unclear. This research investigated the impairment of mammary tissue induced by ZnO NPs and its potential mechanisms. Through administering multiple injections of ZnO NPs into the tail vein of lactating ICR mice, our study revealed that ZnO NPs can deposit in the mammary tissues, downregulating key components of mammary epithelial barrier such as ZO-1, occludin, and claudin-3. In vivo, we also found that ZnO NPs can simultaneously induce apoptosis, necroptosis, and pyroptosis, called PANoptosis. Additionally, using EpH4-Ev cells to simulate an in vitro mammary epithelial barrier model, we observed that ZnO NPs effectively disrupted the integrity of mammary epithelial barrier and induced PANoptosis. Furthermore, we confirmed that PANoptosis was responsible for the mammary epithelial barrier disruption induced by ZnO NPs. Moreover, we identified that ZBP1 was the primary mechanism of ZnO NPs inducing PANoptosis. These discoveries are designed to enhance our comprehension of the mechanisms underlying mammary epithelial barrier disruption caused by ZnO NPs, and we aim to highlight the potential hazards associated with daily usage and therapeutic exposure to ZnO NPs during lactation., 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 Inc. All rights reserved.)

Introduction: Breastfeeding is crucial in infant nutrition and bonding, recommended for at least 6 months and up to 2 years postpartum, providing health benefits for both infants and mothers. However, nipple trauma is common among lactating mothers. Moisturising therapies, such as hydrogel application, aim to promote the healing of nipple wounds. Despite various interventions proposed, comprehensive reviews evaluating the efficacy of moisturising therapies for nipple trauma are lacking. This review aims to appraise the effectiveness of moisturising therapy, particularly tailored for nipple trauma in lactating women., Methods and Analysis: This systematic review protocol adheres to the Preferred Reporting Items for Systematic Review and Meta-Analysis Protocols guidelines and is registered in the International Prospective Register of Systematic Reviews. The focus will be on both randomised and non-randomised controlled trials addressing the treatment of nipple trauma in lactating women. In this context, 'moisturising therapy' is categorised as a non-pharmacological intervention applied directly to the nipple, excluding treatments for bacterial or fungal infections. The primary outcomes will include measures of the healing efficiency of nipple trauma and reduction in nipple pain. Searches will be conducted without date restrictions across multiple databases, including PubMed, CINAHL, the Cochrane Library, Web of Science, CiNii Research and Ichushi-Web, in addition to exploration of grey literature. Two independent reviewers will manage all stages of the review process, under the supervision of a third reviewer. Data extraction will encompass participant demographics, study methodologies, sample specifics and measured outcomes. Quality assessment will be conducted using the Joanna Briggs Institute Appraisal Checklist Tools., Ethics and Dissemination: Ethics approval is not required for this study. The findings of this review will be disseminated through academic publications, detailed reports and presentations at relevant conferences., Prospero Registration Number: CRD42023481761., Competing Interests: Competing interests: None declared., (© Author(s) (or their employer(s)) 2024. Re-use permitted under CC BY-NC. No commercial re-use. See rights and permissions. Published by BMJ.)

Background: The microbiota and metabolites in the gastrointestinal tracts of female animals at different reproductive periods are very important to the growth, development, and health of themselves and their offspring. However, the changes in the gastrointestinal microbiota and metabolites throughout reproductive period of different sheep breeds and their effects on the growth and development of offspring lambs are still unclear. Hence, this study presents an assessment of the reproductive hormone levels, immune levels, rumen microbiota, and metabolites in Hu sheep and Suffolk ewes at different reproductive periods and their effects on the growth and development of offspring lambs., Results: Hu sheep and Suffolk during non-pregnancy, pregnancy, and lactation were used as the research objects to determine reproductive and immune indexes of ewes at different periods, analyze rumen microbiome and metabolome, and track the growth performance and development of offspring lambs. The results showed that the reproductive hormone and immune levels of Hu sheep and Suffolk underwent adaptive changes across different reproductive periods. Compared with non-pregnancy, the microbial energy metabolism and lipid metabolism function decreased during Hu sheep pregnancy, and energy metabolism function decreased during lactation. In Suffolk, energy metabolism, glycan biosynthesis, and metabolism function were enhanced during pregnancy, and the metabolism of cofactors and vitamins was enhanced during lactation. Prevotella increased in Suffolk during pregnancy and lactation (P < 0.05) and was positively correlated with the birth weight and body size of the lambs (P < 0.05). Moreover, the abundances of Butyrivibrio and Rikenellaceae&#95;RC9&#95;gut&#95;group during pregnancy were positively correlated with the intestinal immunity of the offspring lambs (P < 0.05), thereby regulating the intestinal immunity level of the lambs. Metabolomic analysis revealed that the protein digestion, absorption, and amino acid metabolism of Hu sheep were enhanced during pregnancy, which provided amino acids for the growth and development of pregnant ewes and fetuses and was significantly correlated with the birth weight, body size, and intestinal immunity of lambs (P < 0.05). Simultaneously, there was an increase in acetate and propionate during the pregnancy and lactation period of both Hu sheep and Suffolk, providing energy for ewes during reproductive period. Moreover, the microbiota during the lactation period was significantly correlated with the milk quality and lambs daily gain (P < 0.05)., Conclusions: This study revealed the characteristic succession changes in the rumen microbiota and its metabolites at different reproductive periods in sheep breeds and their regulation of reproductive hormone and immune levels and identified their potential effects on the growth and development of offspring lambs. The findings provide valuable insights into the health and feeding management of different sheep breeds during the reproductive stage. Video Abstract., (© 2024. The Author(s).)

Sarcopenia is characterised by the loss of skeletal muscle mass and function, which leads to a high risk of increased morbidity and mortality. Maternal malnutrition has been linked to impaired development of skeletal muscle of the offspring; however, there are limited studies that report the long-term effect of a maternal low-protein diet during lactation on the ageing of skeletal muscles. This study aimed to examine how a maternal low-protein diet (LPD) during lactation affects skeletal muscle ageing in the offspring. Pups born from control mothers were lactated by mothers fed with an LPD. Post-weaning, mice were either maintained on an LPD or switched to a control, normal-protein diet (NPD). In males, an LPD mainly affected the size of the myofibres without a major effect on fibre number and led to reduced grip strength in ageing mice (24 months). Female mice from mothers on an LPD had a lower body and muscle weight at weaning but caught up with control mice at 3 months. During ageing, the muscle weight, myofibre number and survival rate of female pups were significantly affected. These findings highlight the effect of an LPD during lactation on skeletal muscle ageing, the lifespan of offspring and the importance of sexual dimorphism in response to dietary challenges.

Cow-calf contact (CCC) rearing is becoming an increasingly popular alternative to the common practice of early separation of cow and calf in dairy management. Milkability can be impaired in nursing cows, which contributes to the loss of machine milk yield caused by calf intake, especially in pure dam-calf contact (DCC) systems. The aims of this study were (1) to describe the current status quo of DCC rearing regarding management and milking and (2) to evaluate the effects of DCC (suckling and milking vs. milking alone) and the effects of different types of DCC on milkability parameters, teat condition, and behavior during milking on Swiss DCC farms. By means of 17 telephone interviews with DCC farmers, we collected data on DCC management, housing, separation and weaning processes, milking procedures and techniques, and perceived milkability problems. Subsequently, we collected data on 10 of the interviewed DCC farms (183 cows): 4 DCC farms with a whole-day contact (WDC) system, 3 farms with DCC before milking (CBM), and 3 farms with DCC after milking (CAM). Five farms on which calves had no contact with dams were chosen as reference farms (178 cows). Using a milk flow meter, the occurrence of ejection disorders, bimodality of the milk flow curve, machine milk yield, duration of the decline phase, and duration of prestimulation were measured. The average mouthpiece chamber vacuum during the main milking phase and hind leg activity during milking were measured using a pressure sensor and an accelerometer, respectively. After cluster removal, the teat condition was evaluated, and a stripping milk sample was taken for fat content analysis. The interview results revealed that 8 of the 17 farms surveyed had a WDC system, and 2 farms operated a daytime DCC system. Contact before milking was applied by 3 farms, and 3 farms allowed CAM. On one farm, calves had access to dams 3 times a day. A great diversity in cow-calf management was found. In the on-farm data collection, 20 milkings of a total of 701 milkings examined met the criteria for a clear ejection disorder, with 17 of these observations occurring on WDC farms and none on reference farms. The stripping milk fat content was lower in nursing cows, indicating a lower degree of udder emptying. Machine milk yield was higher in nursing CAM cows than in nursing WDC and CBM cows. Farm types did not differ regarding teat condition, hind leg activity, or the occurrence of bimodal milk flow curves. In conclusion, the large variation in individual management approaches to DCC rearing even within DCC types, such as calf housing or cow breeds, implies caution when interpreting results. Contact after milking may be the system most beneficial for some productivity parameters, but adequate calf supply must be ensured. Higher amounts of milk remaining in the udder after cluster removal indicate that nursing can affect milkability, but future research should consider the effects of udder filling before milking to better interpret the fat content of stripping milk., (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 investigated the short- and long-term effects of different forage types supplemented in preweaning dairy calves on growth performance, blood metabolites, rumen fermentation, bacterial community, and milk production during first lactation. A total of 60 healthy 30-d-old female Holstein calves were blocked by birth date and body weight and randomly assigned to 1 of 3 groups (n = 20): normal milk and pelleted starter feeding (CON), supplemented with chopped oat hay (OAH; 75.0 g/d per calf [DM basis]), or alfalfa hay (ALF; 75.0 g/d per calf [DM basis]). The forage supplementation started when calves were 30 d old (d 1 of the experimental period) and ended when they were 73 d old (d 44 of the experimental period, when calves were weaned). Milk and feed intakes and fecal consistency scores were recorded daily. Growth performance, rumen fluid, and blood samples were collected biweekly. After weaning, all the calves were integrated with the same barn and diets. After calving, the milk production was recorded daily. During the experimental period, the OAH group had greater solid feed and total DM intakes and greater rumen pH than the CON group (P ≤ 0.04), but had lower forage intake and CP digestibility than the ALF group (P ≤ 0.04). The ALF group had higher rumen pH and blood BHB concentration (P ≤ 0.04), lower fecal score (P = 0.02), and greater ether extract digestibility (P = 0.02) than the CON group. The ALF and OAH groups had lower concentrations of ruminal total VFA (P = 0.01). Still, the ALF group had a greater proportion of acetate and a relative abundance of cellulose degradation-related bacteria (Lachnoclostridium&#95;1 and Oribacterium) and a lower relative abundance of inflammation-related bacteria (Erysipelotrichaceae&#95;UCG-009) in the rumen compared with CON. Interestingly, the average milk production from 6 to 200 DIM was greater in the ALF group (P < 0.01), even though no significant effects were found on the rumen fermentation parameters and blood metabolites at 200 DIM. Generally, alfalfa hay supplementation in preweaning dairy calves had positive effects in the short- and long-term for rumen development, health status, and future milk production., (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: Concentrations of vitamin D (VitD) and 25-hydroxyvitamin D (25OHD) in breastmilk are low despite the essential role of VitD for normal infant bone development, yet additional metabolic forms of vitamin D may be present. This study evaluates the contribution of sulfated vitamin D metabolites, vitamin D 3 -sulfate (VitD 3 -S) and 25-hydroxyvitamin D 3 -sulfate (25OHD 3 -S) for lactating women and assesses the response to high-dose VitD 3 supplementation., Methods: Serum and breastmilk were measured before and after 28 days with 5000 IU/day VitD 3 intake in 20 lactating women. Concentrations of VitD 3 -S and 25OHD 3 -S in milk, and 25OHD 2 , 25OHD 3 , 25OHD 3 -S, VitD 3 and VitD 3 -S in serum were determined by mass spectrometry., Results: Baseline vitamin D status was categorized as sufficient (mean ± SD serum 25OHD 3 69 ± 19 nmol/L), and both serum VitD 3 and 25OHD 3 increased following supplementation (p < 0.001). 25OHD 3 -S was 91 ± 19 nmol/L in serum and 0.47 ± 0.09 nmol/L in breastmilk. VitD 3 -S concentrations were 2.92 ± 0.70 nmol/L in serum and 6.4 ± 3.9 nmol/L in breastmilk. Neither sulfated metabolite significantly changed with supplementation in either serum or breastmilk., Conclusions: Sulfated vitamin D metabolites have prominent roles for women during lactation with 25OHD 3 -S highly abundant in serum and VitD 3 -S distinctly abundant in breastmilk. These data support the notion that 25OHD 3 -S and VitD 3 -S may have physiological relevance during lactation and nutritional usage for nursing infants., Competing Interests: Conflict of interest The authors have no commercial or financial conflicts of interest to declare., (Copyright © 2024 The Authors. Published by Elsevier Ltd.. All rights reserved.)

A dysregulated inflammatory response contributes to the occurrence of disorders in cows during the transition period from pregnancy to lactation. However, a detailed characterization of clinically healthy cows that exhibit an enhanced inflammatory response during this critical period remains incomplete. In this experiment, a total of 99 individual transition dairy cows and 109 observations (18 cows monitored in 2 consecutive lactations), submitted to similar transition management were involved to evaluate the relationship between elevated an inflammatory response and metabolic and oxidative status, as well as transition outcomes. Blood was taken at -7, 3, 6, 9, and 21 DIM, and concentrations of metabolic parameters (glucose, β-hydroxybutyric acid, nonesterified fatty acids [NEFA], insulin, IGF-1, and fructosamine) were analyzed. Additionally, oxidative parameters (proportion of oxidized glutathione to total glutathione in red blood cells, the activity of glutathione peroxidase [GPx] and superoxide dismutase, concentrations of malondialdehyde, and oxygen radical absorbance capacity) and acute phase proteins (APP) including haptoglobin (Hp), serum amyloid A (SAA) and albumin-to-globulin ratio (A:G) were determined in the blood at 21 DIM. The 3 APP parameters were used to group clinically healthy cows into 2 categories through k-medoids clustering (i.e., a group showing an acute phase response, APR; n = 39) and a group not showing such a response (i.e., non-APR; n = 50). Diseased cases (n = 20) were handled in a separate group. Lower SAA and Hp concentrations as well as higher A:G were observed in the non-APR group, although for Hp, differences were observed from the APR group and not from the diseased group. Only 1 of the 5 oxidative parameters differed between the groups, with the non-APR group exhibiting lower GPx activity compared with the diseased group. The non-APR group showed the highest IGF-1 levels among the 3 groups and and lower NEFA concentrations compared with the diseased groups. Cows in the diseased group also showed reduced dry matter intake and milk yield compared with clinically healthy cows, regardless of their inflammatory status. Moreover, the APR group exhibited temporarily lower activity levels compared with the non-APR group. These findings highlight that cows with a lower inflammatory status after 21 DIM exhibited better metabolic health characteristics and productive performance, as well as activity levels. Nevertheless, the detrimental effects of a higher inflammatory status in the absence of clinical symptoms are still relatively limited., (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/).)

Reduced feed intake in early lactation prompts increased fat mobilization to meet dairy cow energy needs for milk production. The increased lipolysis in cows presents significant health risks with unclear mechanisms. The objectives of our study were to compare the longitudinal profiles of metabolites and lipids of serum from high- and low-lipolysis cows. Forty multiparous Holstein dairy cows were enrolled in the retrospective study. Serum samples were collected on d 7 before expected calving, as well as on d 5, 7, 14, and 21 postpartum. Dairy cows were grouped according to mean serum nonesterified fatty acids on d 5 and 7 after parturition as low (<0.600 mmol/L; n = 8; LFM) and high (>0.750 mmol/L; n = 8; HFM), indicating fat mobilization during early lactation. Lactational performance and serum metabolic parameters related to glucose and lipid metabolism, liver functions, oxidative status, and inflammatory responses were determined. Serum samples were subjected to liquid chromatography-MS-based metabolomics and lipidomics. Despite differences in postpartum BW change, there were no observed variations in milk yield and composition between the 2 groups. Serum β-hydroxybutyric acid, glucose, leptin, aspartate aminotransferase, IL-6, and tumor necrosis factor alpha were greater in cows with HFM than in cows with LFM. Serum adiponectin, revised quantitative insulin sensitivity check index, and albumin were lower in cows with HFM than in cows with LFM. Intensified fat mobilization in the HFM cows came along with reduced estimated insulin sensitivity, impaired liver functions, and increased oxidative stress and inflammatory responses. Differences in metabolic patterns were observed across the transition period when comparing serum blood matrixes (e.g., in different amino acids, acylcarnitines, and sphingolipids). The serum metabolome of the HFM cows was characterized by higher concentrations of glycine, acylcarnitines, carnosine, Cer(d20:0/18:0), Cer(d18:1/16:0), and Cer(t18:0/24:0) compared with LFM cows. The differential serum metabolites and lipids at different sampling times during the peripartum period were enriched in the sphingolipid metabolism. Differences in serum metabolic status parameters suggest that cows adopt varied metabolic adaptation strategies to cope with energy deficits postpartum. Our investigation found a comprehensive remodeling of the serum metabolic profiles in transition dairy cattle, highlighting the significance of alterations in sphingolipid species, as they play a crucial role in insulin resistance and metabolic disorders., (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 assess the effect of a reduced liner-open phase applied through flow-responsive pulsation (FRP), as a method to provide supplemental stimulation, on teat tissue conditions and milking characteristics in dairy cows. In 2 switch-back trials, 156 Holstein cows milked 3 times daily were assigned to the FRP or conventional (CON) group in alternating sequences. Trial 1 lasted for 35 d and was split into 5 alternating 1-wk periods of FRP and CON. The duration of trial 2 was 84 d, consisting of 4 alternating 3-wk periods of FRP and CON. Premilking udder preparation for both groups consisted of predipping, forestripping, and wiping the teats. Upon milking unit attachment, the FRP cows were milked at a pulsation rate of 50 cycles/min and a pulsation ratio of 30:70 until the preset milk flow threshold of 0.5 kg/min was reached. When the threshold value of 0.5 kg/min was reached, the pulsation was automatically switched to milking mode, which consisted of a pulsation rate of 60 cycles/min and a pulsation ratio of 70:30. Cows in the CON group were milked by milking mode (pulsation rate, 60 cycles/min; pulsation ratio, 70:30) immediately after attachment of the milking unit. We assessed machine milking-induced short-term changes to the teat tissue by palpation and visual inspection during trial 1, and we assessed teat-end hyperkeratosis in trial 2. Electronic on-farm milk meters were used to assess milking characteristics: milk yield (kg/milking session), milking unit-on time (s), 2-min milk yield (kg), peak milk flow rate (kg/min), and duration of low milk flow rate (s). Generalized linear mixed models were used to analyze the effects of treatment on the outcome variables. The odds of machine milking-induced short-term changes to the teat tissue were lower for cows in the FRP group than for those in the CON group: odds ratio (95% CI) = 0.41 (0.31-0.55). There were no meaningful differences in the odds of teat-end hyperkeratosis between the FRP and CON groups: odds ratio (95% CI) = 1.05 (0.38-2.89). The LSM (95% CI) of milking characteristics in the FRP and CON groups were 14.3 (13.8-14.7) and 14.3 kg (13.8-14.7) for milk yield, respectively; 272 (264-281) and 270 s (262-278) for milking unit-on time, respectively; and 5.0 (4.8-5.1) and 4.9 kg/min (4.7-5.1) for peak milk flow rate, respectively. The FRP group had lower odds of bimodality than the CON group: odds ratio (95% CI) = 0.67 (0.61-0.74). In reference to CON, the odds ratios (95% CI) in FRP were 1.05 (0.76-1.46) for kick-off and 1.02 (0.85-1.23) for milking unit reattachment. In this study, cows that were milked using FRP had lower odds of postmilking short-term changes to the teat tissue and lower odds of bimodal milk flow. We conclude that FRP may foster adequate teat stimulation in cows before the initiation of milk harvest and has the potential to improve teat tissue conditions., (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 determine quarters requiring antimicrobial treatment using either a benchtop somatic cell counter or culture with gram-positive selective media and compare the outcomes in these cows to those receiving blanket dry cow therapy (BDCT) in a randomized, controlled trial. We evaluated 2 novel methods of identifying cows with intramammary infections followed by selective antimicrobial treatment at a commercial dairy farm to determine their usefulness in decreasing antibiotic usage during the dry period without significant detrimental effects on milk quality and production. Cows (n = 840) were randomly allocated to one of 3 groups (BDCT, gram-positive selective media culture-based selective dry cow therapy [C-SDCT], and somatic cell count-based SDCT [S-SDCT]) the day before dry-off, and quarter-level milk samples (QLMS) were collected. The QLMS from cows in the S-SDCT group were evaluated using the cell counter, and quarters were treated if SCC was ≥200,000 cells/mL, whereas the QLMS from cows in the C-SDCT group were cultured, and quarters were treated if the culture showed growth. All cows in the BDCT received antimicrobial therapy, and all cows received an internal teat sealant regardless of treatment group. Outcomes measured were first and second DHIA test SCC, milk production through 60 DIM, cows leaving the farm, clinical mastitis, and bacteriologic new infections in a subset of quarters. Cows in both SDCT groups had fewer antimicrobial treatments than cows in the BDCT group as was expected, and cows in the C-SDCT group had fewer treatments than those in the S-SDCT group. Cows in both SDCT groups had a higher linear score at the first DHIA test (BDCT: 1.8, S-SDCT: 2.2, C-SDCT: 2.2); however, we found no other differences between groups regarding any other outcomes measured. Although antimicrobial use was significantly reduced, farms should use caution in adopting the benchtop analyzer and the selective media described in this study as ways to identify infected cows for dry cow therapy because they may result in increased linear scores early in 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/).)

Researchers have reported the benefits of feeding rumen-protected methionine (RPM) during the peripartum on the health parameters of dairy cows. Rumen-protected Met has reportedly improved milk yield, milk components, and liver health, but the literature is scarce on its effects in commercial herds. Therefore, we aimed to determine the effects of feeding RPM (Smartamine M, Adisseo Inc., Antony, France) prepartum (8 g/cow per day) and postpartum (15 g/cow per day) on performance, metabolic profile, and culling rate of Holstein cows in a commercial herd. One hundred sixty-six (n = 166) Holstein cows, 58 nulliparous and 108 parous, were randomly assigned to 1 of 2 dietary treatments, consisting of TMR top-dressed with RPM (RPMet; 2.35% and 2.24% Met of MP for close-up and fresh cows, respectively) or without (control [CON] 2.03% and 1.89% Met of MP for close-up and fresh cows, respectively), fed from 21 ± 6 d prepartum until 16 ± 5 d postpartum. From 17 DIM until dry-off, all cows received RPMet. Daily milk yield was recorded, and milk samples were collected in the first and second weeks after calving to determine their composition. Blood samples were collected before the morning feeding on -14, -7, +1, +7, and +14 d relative to calving. Mortality and morbidity were recorded during the first 60 DIM. Cows supplemented with RPMet had greater milk yield during the first 16 DIM (31.76 vs. 30.37 kg/d; SEM = 1.04, respectively), and had greater milk fat content (4.45 vs. 4.10%; SEM = 0.11, respectively), but not milk total protein (3.47 vs. 3.39%; SEM = 0.04, respectively) and casein contents (2.74 vs. 2.66%; SEM = 0.04, respectively) than CON cows. Cows in RPMet had increased plasma Met concentrations than cows in CON (24.9 vs. 21.0 µmol/L; SEM = 1.2, respectively). Although morbidity was similar between treatments, the culling rate from calving until 60 DIM was lower for RPMet cows than for CON cows (2.4% vs. 12.1%; SEM = 0.02). In conclusion, cows receiving RPMet have greater milk yield, improved milk fat content, and a lower culling rate at 60 DIM than CON 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/).)

Flaxseed is the richest source of secoisolariciresinol diglucoside, which is converted by ruminal microorganisms primarily to the mammalian lignan enterolactone. Our objective was to investigate the effect of diets containing soybean meal or flaxseed meal (FM) supplemented with sucrose, flaxseed oil, or both, on milk enterolactone concentration yield, diversity, and relative abundance of ruminal bacterial taxa, ruminal fermentation profile, production performance, milk fatty acids (FA) yield, and nutrient utilization in dairy cows. Sixteen Holstein cows (8 multiparous [4 ruminally cannulated] and 8 primiparous) averaging (mean ± SD) 134 ± 54.1 DIM and 679 ± 78.9 kg of BW in the beginning of the study were assigned to treatment sequences in a replicated 4 × 4 Latin square design. Each experimental period lasted 25 d, with 18 d for diet adaptation and 7 d for data and sample collection. Diets were formulated to contain a 60:40 forage:concentrate ratio and included the following (DM basis): (1) 8% soybean meal and 23% ground corn (SBM); (2) 15% FM, 10.7% ground corn, and 5% sucrose (FLX+S); (3) 15% FM, 15.4% ground corn, and 3% flaxseed oil (FLX+O); and (4) 15% FM, 10.2% ground corn, 5% sucrose, and 3% flaxseed oil (FLX+SO). Compared with SBM, the concentration and yield of milk enterolactone increased in cows fed the FM diets but did not differ among FLX+S, FLX+O, and FLX+SO. The relative abundances of the phyla Firmicutes, Verrucomicrobiota, and Actinobacteriota, and those of the bacterial genera Lachnospiraceae NK3A20 group, Eubacterium coprostanoligenes group, Anaeromusa-Anaeroarcus, WCHB1-41, and p-251-o5 decreased, whereas Prevotella and NK4A214 group increased when comparing SBM against at least 1 diet containing FM. Furthermore, the relative abundances of Firmicutes and Actinobacteriota and those of Prevotella, Lachnospiraceae NK3A20 group, Eubacterium coprostanoligenes group, Acetitomaculum, Lachnospiraceae unclassified, NK4A214 group, and Anaeromusa-Anaeroarcus changed (increased or decreased) across the FLX+S, FLX+O, and FLX+SO diets. However, all these changes in the relative abundance of the ruminal bacterial taxa were not conclusively associated with the effect of diets on milk enterolactone. Diets did not affect ruminal pH and concentrations of NH 3 -N and total VFA. Dry matter intake and yields of milk, milk fat, and milk true protein all decreased in cows fed FLX+O or FLX+SO. Yields of milk total odd-chain FA, branched-chain FA, total <16C FA, and total 16C FA decreased with feeding FLX+O and FLX+SO. The apparent total-tract digestibilities of DM and OM were lowest in the FLX+S and FLX+O diets, with CP and ADF digestibilities lowest in cows receiving FLX+S or FLX+O, respectively. Urinary excretion of total N was lowest with feeding SBM. Contrarily, diets did not affect the urinary excretion of total purine derivatives. In brief, despite the effect of diets on the relative abundance of several ruminal microbiota phyla and genera, we were unable to conclusively associate these changes with increased milk enterolactone in FM-containing diets versus SBM., (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/).)

Automated measurements of the ratio of concentrations of methane and carbon dioxide, [CH 4 ]:[CO 2 ], in breath from individual animals (the so-called "sniffer technique") and estimated CO 2 production can be used to estimate CH 4 production, provided that CO 2 production can be reliably calculated. This would allow CH 4 production from individual cows to be estimated in large cohorts of cows, whereby ranking of cows according to their CH 4 production might become possible and their values could be used for breeding of low CH 4 -emitting animals. Estimates of CO 2 production are typically based on predictions of heat production, which can be calculated from body weight (BW), energy-corrected milk yield, and days of pregnancy. The objectives of the present study were to develop predictions of CO 2 production directly from milk production, dietary, and animal variables, and furthermore to develop different models to be used for different scenarios, depending on available data. An international dataset with 2,244 records from individual lactating cows including CO 2 production and associated traits, as dry matter intake (DMI), diet composition, BW, milk production and composition, days in milk, and days pregnant, was compiled to constitute the training dataset. Research location and experiment nested within research location were included as random intercepts. The method of CO 2 production measurement (respiration chamber [RC] or GreenFeed [GF]) was confounded with research location, and therefore excluded from the model. In total, 3 models were developed based on the current training dataset: model 1 ("best model"), where all significant traits were included; model 2 ("on-farm model"), where DMI was excluded; and model 3 ("reduced on-farm model"), where both DMI and BW were excluded. Evaluation on test dat sets with either RC data (n = 103), GF data without additives (n = 478), or GF data only including observations where nitrate, 3-nitrooxypropanol (3-NOP), or a combination of nitrate and 3-NOP were fed to the cows (GF+: n = 295), showed good precision of the 3 models, illustrated by low slope bias both in absolute values (-0.22 to 0.097) and in percentage (0.049 to 4.89) of mean square error (MSE). However, the mean bias (MB) indicated systematic overprediction and underprediction of CO 2 production when the models were evaluated on the GF and the RC test datasets, respectively. To address this bias, the 3 models were evaluated on a modified test dataset, where the CO 2 production (g/d) was adjusted by subtracting (where measurements were obtained by RC) or adding absolute MB (where measurements were obtained by GF) from evaluation of the specific model on RC, GF, and GF+ test datasets. With this modification, the absolute values of MB and MB as percentage of MSE became negligible. In conclusion, the 3 models were precise in predicting CO 2 production from lactating dairy 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/).)

The presence of an autocrine factor in milk that can trigger mammary gland involution was proposed more than 50 yr ago. To provide evidence for the existence of one or more autocrine factors, 10 multiparous cows in late lactation were quarter-milked for 7 d. Following this baseline period, the right front quarter of each cow was left unmilked, and the other quarters were milked for 7 d. Before the last milking of that period, milk (mammary secretions) was collected aseptically from both front quarters. After that milking, 250 mL of the collected samples were infused in the cows' respective rear quarters. No quarters were milked for the following 7 d (milk stasis period), and quarter milking was then resumed in all quarters for the last 7 d of the experiment (remilking period). Quarter milk samples were collected during the baseline period, before the milk stasis period, and during the remilking period. These samples were used for measuring milk components and the concentration of involution markers (SCC, BSA, and lactoferrin). Samples of mammary secretions were collected manually from the quarters during the milk stasis period for involution marker determination. We extracted RNA from samples collected from front quarters before the last milking before the milk stasis period for microRNA (miRNA) determination. As anticipated, the longer milk stasis period implemented for the right front quarter resulted in a more advanced involution than in the left front quarter, based on the concentration of involution markers in the mammary secretions, lower milk production recovery, and changes in milk composition during the remilking period. All 3 involution marker concentrations in the mammary secretions increased in both rear quarters, but were greater in the right quarter secretions than in the left quarter secretions. Resuming milking reinitiated milk production in all quarters, but milk production recovery in the right rear quarters was less robust than that in the left rear quarters (54.3 ± 1.4% vs. 61.6 ± 1.4%, respectively). Milk from the quarters infused with mammary secretions (right rear) had a lower lactose content, but a higher milk protein content and higher SCC than the quarters infused with milk. We detected a total of 359 miRNAs, 76 of which were differentially expressed in milk and mammary secretions. Expression of bta-miR-221 and bta-miR-223 was upregulated in mammary secretions 34- and 40-fold, respectively. The results of the present experiment support the contention that milk stasis leads to the accumulation of one or more factors that trigger involution. The results also indicate that milk stasis leads to changes in the miRNA profile of the milk, but whether such changes are a cause or a consequence of the involution process remains to be established., (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/).)

Microplastics have become a ubiquitous contaminant, but their fate in food animals is largely unknown. In this study, [ 14 C]-polystyrene microplastic (PS-MP) particles were orally dosed to lactating sheep to evaluate their absorption and disposition. Elimination of the [ 14 C]-PS-MP was predominately through faeces with faecal radioactivity peaking at 24 h post-dosing but continuing to be present throughout the entire 72 h study period. Only a small fraction (≤ 1%) of the dosed [ 14 C]-PS-MP was present in blood, milk, and urine. Pharmacokinetic analysis of blood plasma radioactivity, using non-compartment modeling, indicated rapid absorption (T 1/2 0.4 to 3 h) with slow elimination (T 1/2 37 to 48 h). Radioactivity in milk and urine had similar elimination patterns with radiocarbon activities peaking 24 h post-dosing with detectable elimination throughout the 72 h study period. No radioactivity was quantifiable in tissues at the 72 h withdrawal period.

Our objective was to estimate the requirements of metabolizable energy (ME) and NE M of lactating and dry cows, the efficiency of ME utilization for milk production (k l ) and tissue gain (k g ), and the use of body energy mobilization for milk production (k t ) throughout the lactation of primiparous crossbred Holstein × Gyr cows, using open-circuit respiration chambers. Twenty-nine primiparous Holstein × Gyr crossbred cows with an initial BW averaging 563 ± 40.1 kg and 2.5 ± 0.09 yr old were used throughout lactation and dry periods. The cows were kept nonpregnant throughout the study to eliminate possible confounding factors. Apparent digestibility assays, followed by calorimeter measurements, were performed 6 times throughout the lactation period. In the dry-off period, the cows were also evaluated but fed with restricted intake (DMI = 1.1% BW/d) to achieve heat production close to maintenance. After 21 d of diet adaptation, an apparent digestibility assay followed by calorimeter measurements was performed. Parameter estimates for lactation period were obtained by mixed models including lactation stage as repeated measures. For restricted feeding at dry-off and fasting period assays, the requirements were estimated by exponential regression. For whole lactation, the values of the ME requirement for maintenance (ME M ) and NE M were 0.588 and 0.395 MJ/BW 0.75 , respectively. The efficiencies of k l , k g , and k t were 0.672, 0.771, and 0.814, respectively. However, ME M and NE M were higher in early and mid lactation than late, whereas k l was higher in early than other lactation stages. Dry and nonpregnant cows had ME M of 0.434 MJ/BW 0.75 and NE M of 0.351 MJ/BW 0.75 for maintenance level, and ME M of 0.396 MJ/BW 0.75 and NE M of 0.345 MJ/BW 0.75 for fasting metabolism level, and efficiency of ME utilization for maintenance was 0.80. Our findings confirmed that F 1 crossbred Holstein × Gyr dairy cows have differences in energy requirement and efficiency throughout the lactation stages, suggesting the use of different values in each stage. The estimated values of energy requirement for maintenance and efficiencies for primiparous lactating crossbred Holstein × Gyr were similar to those reported in the literature in specific studies and requirements systems., (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 AA profile of MP affects mammary gland metabolism and milk N efficiency of dairy cattle. Further, the frequency of dietary protein supplementation may influence N partitioning leading to reduced N excretion. This study investigated the effect of source and frequency of rumen-protected (RP) protein supplementation on apparent total-tract digestibility, milk production, mammary gland AA metabolism, and N balance of dairy cattle. Twenty-eight Holstein-Friesian cows (2.3 ± 0.9 lactations; 93 ± 27 DIM; mean ± SD) were used in a randomized complete block design and fed a basal TMR consisting of 41% corn silage, 32% grass silage, and 27% concentrate (DM basis) and formulated to meet 100% and 95% of net energy and MP requirements, respectively. Cows were adapted to the basal TMR in a freestall barn for 7 d, moved to individual tiestalls for 13 d of adaptation to dietary treatments, and then moved into climate respiration chambers for a 4-d measurement period. Treatments consisted of the basal TMR (CON; 159 g CP/kg DM) or the basal TMR including 1 of 3 iso-MP supplements: (1) 315-g mixture of RP soybean meal and RP rapeseed meal fed daily (ST-RPSR), (2) 384-g mixture of RP His, RP Lys, and RP Met fed daily (ST-RPAA), and (3) 768-g mixture of RP His, RP Lys, and RP Met fed every other day (OS-RPAA). The basal TMR with the addition of treatment supplements was designed to deliver 100% of required MP over a 48-h period. The mixture of His, Lys, and Met was formulated to deliver digestible AA in amounts relative to their concentration in casein. Compared with ST-RPSR, ST-RPAA increased milk protein and fat concentration, increased the arterial concentration of total His, Lys, and Met (HLM), decreased mammary clearance of HLM, and increased clearance of Phe, Leu, and Tyr (tendency for Leu and Tyr). Rumen-protected protein source did not affect N balance, but the marginal use efficiency (efficiency of transfer of RP protein supplement into milk protein) of ST-RPAA (67%) was higher than that of ST-RPSR (17%). Milk protein concentration decreased with OS-RPAA compared with ST-RPAA. Arterial concentration of HLM increased on the nonsupplemented day compared with the supplemented day with OS-RPAA, and there was no difference in arterial HLM concentration across days with ST-RPAA. Mammary uptake of HLM tended to increase on the nonsupplemented day compared with the supplemented day with OS-RPAA. Supplementation frequency of RP AA did not affect N balance or overall milk N efficiency, but the marginal use efficiency of OS-RPAA (49%) was lower compared with ST-RPAA. Overall, mammary glands responded to an increased supply of His, Lys, and Met by reducing efflux of other EAA when RP His, RP Lys, and RP Met were supplemented compared with RP plant proteins. Mammary glands increased sequestration of EAA (primarily HLM) on the nonsupplemented day with OS-RPAA, but supplementing RP AA according to a 24-h oscillating pattern did not increase N efficiency over static supplementation., (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/).)

Objective: Wound healing is a complex and dynamic process essential for restoring tissue integrity and homeostasis. It is thought that breast milk contributes positively to the wound healing process, thanks to the components it contains. The aim of this study is to compare the effects of breast milk on the wound healing process at different lactation stages and to evaluate the underlying mechanism(s). Materials and Methods: The effects of breast milk from different lactation stages (colostrum, transitional, and mature milk) on wound healing were determined by in vitro scratch assay in L929 fibroblast cells. 2,2-Diphenyl-1-picrylhydrazyl (DPPH), total oxidant, and antioxidant capacity were used to confirm antioxidant effects. The effect of breast milk on netrin-1 levels in L929 cells was elucidated by ELISA. Results: Breast milk at different lactation stages promoted wound healing. While the wound closure percentage was determined as 48.7% in the control group, this rate was determined to be the highest at 81.6% in the mature milk group (p:0.0002). The free radical scavenging capacity of colostrum, transitional, and mature milk with DPPH was determined as 49.69%, 60.64%, and 80.85%, respectively, depending on the lactation stages. Netrin-1 levels detected by ELISA were determined as 490.1 ± 6.5 pg/mL in the control group, while the lowest level was determined as 376.6 ± 4.5 pg/mL in mature milk (p:0.0003). Conclusions: Breast milk, especially mature milk, promoted wound healing on L929 cells by suppressing netrin-1 levels and scavenging free radicals.

Urine output and urinary urea-N excretion (UUN e ) excretion are critical measures to accurately evaluate N metabolism in lactating dairy cows and environmental concerns related to manure N. The objectives of this study were: (1) to compare estimates of UUN e , urine output, and related variables from 3 pre-established measurement protocols (bladder catheterization, external collection cup, and spot sampling) and from dietary salt supplementation, (2) to study temporal variation in UUN e , urine output, and related variables as affected by measurement protocol, and (3) to evaluate urine specific gravity as a predictor of urine output. Twelve multiparous Holstein cows were used in a split-plot, Latin square design. Cows were randomly assigned to a diet (main plot) containing either 0.7% or 1.6% NaCl (DM basis) and then assigned to a sequence of 3 protocols (subplot) in a balanced 3 × 3 Latin square with 14-d period. For each protocol, measurements were conducted every 4 h for 3 consecutive days. Urine output was determined gravimetrically for bladder catheterization and external collection cup or based on measured cow BW, measured urinary creatinine concentration, and the assumed creatinine excretion of 29 mg/kg BW per day for spot sampling. Urine specific gravity was measured by refractometry. When averaged over a 3-d measurement period and compared with bladder catheterization, spot sampling underestimated urine output (6.8 kg/d; 20%) and UUN e (26 g/d; 13%) but exhibited greater concentration of urinary urea-N (+58 mg/dL; 10%). There were no differences in any measurements determined via bladder catheterization or external cup device protocols, except for urine output that tended to be 3.7 kg/d lower for collection cup compared with bladder catheterization. The 2 gravimetric protocols yielded lower urinary creatinine concentration than spot sampling (64.7 vs. 88.1 mg/dL) and lower creatinine excretion (25.3 mg/kg BW per day) than the value of 29 mg/kg BW per day generally assumed in the spot sampling protocol. Salt supplementation tended to increase urine output (+5.2 kg/d) and decrease urinary urea-N concentration (-93 mg/dL), urinary creatinine concentration (-9.5 mg/dL), milk protein concentration (-0.19 percentage unit) and milk protein yield (-70 g/d). There was greater temporal variation of urine output when measured via the collection cup compared with bladder catheterization in the first 2 d but not the third day of sampling, suggesting that an extended period of adaptation might have improved data quality of the collection cup protocol. The R 2 of the linear regression to predict urine output with urine specific gravity was 67%, 73%, and 32% for bladder catheterization, collection cup, and spot sampling, respectively. In this study, spot sampling underestimated both urine output and UUN e , but UUN e determination did not differ between external collection cup and bladder catheterization. However, our data suggested the need to investigate the adaptation protocol, required days of measurements and the conversion of urine mass to urine volume to improve accuracy and precision of urine collection protocols., (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/).)

Large datasets allow estimation of feed required for individual milk components or body maintenance. Phenotypic regressions are useful for nutrition management, but genetic regressions are more useful in breeding programs. Dry matter intake records from 8,513 lactations of 6,621 Holstein cows were predicted from phenotypes or genomic evaluations for milk components and body size traits. The mixed models also included DIM, age-parity subclass, trial date, management group, and BW change during 28- and 42-d feeding trials in mid lactation. Phenotypic regressions of DMI on milk (0.014 ± 0.006), fat (3.06 ± 0.01), and protein (4.79 ± 0.25) were much less than corresponding genomic regressions (0.08 ± 0.03, 11.30 ± 0.47, and 9.35 ± 0.87, respectively) or sire genomic regressions multiplied by 2 (0.048 ± 0.04, 6.73 ± 0.94, and 4.98 ± 1.75). Thus, marginal feed costs as fractions of marginal milk revenue were higher from genetic than phenotypic regressions. According to the ECM formula, fat production requires 69% more DMI than protein production. In the phenotypic regression, it was estimated that protein production requires 56% more DMI than fat. However, the genomic regression for the animal showed a difference of only 21% more DMI for protein compared with fat, whereas the sire genomic regressions indicated approximately 35% more DMI for fat than protein. Estimates of annual maintenance in kilograms DMI/kilograms BW per lactation were similar from phenotypic regression (5.9 ± 0.14), genomic regression (5.8 ± 0.31), and sire genomic regression multiplied by 2 (5.3 ± 0.55) and are larger than those estimated by the National Academies for Science, Engineering, and Medicine based on NE L equations. Multiple regressions on genomic evaluations for the 5 type traits in body weight composite (BWC) showed that strength was the type trait most associated with BW and DMI, agreeing with the current BWC formula, whereas other traits were less useful predictors, especially for DMI. The Net Merit formula used to weight different genetic traits to achieve an economically optimal overall selection response was revised in 2021 to better account for these estimated regressions. To improve profitability, breeding programs should select smaller cows with negative residual feed intake that produce more milk, fat, and protein., (Copyright © 2024 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

The objective was to compare differences in reproductive performance for dairy cows grouped based on the combination of data for predictors available during the prepartum period and before the end of the voluntary waiting period (VWP), automated estrus alerts (AEA) during the VWP, and the combination of both factors. In a cohort study, data for AEA and potential predictors of the percentage of cows that receive AI at detected estrus (AIE), pregnancies per AI (P/AI) for first service, and the percentage of cows pregnant by 150 DIM (P150) were collected from -21 to 49 DIM for lactating Holstein cows (n = 886). The association between each reproductive outcome with calving season (cool, warm), calving-related events (yes, no), genomic daughter pregnancy rate (gDPR; high, medium, low), days in the close-up pen (ideal, not ideal), health disorder events (yes, no), rumination time (high or low CV prepartum and high or low increase rate postpartum), and milk yield (MY) by 49 DIM (high, medium, low) were evaluated in univariable and multivariable logistic regression models. Individual predictors (health disorders, gDPR, and MY) associated with the 3 reproductive outcomes in all models were used to group cows based on risk factors (RF; yes, n = 535 or no, n = 351) for poor reproductive performance. Specifically, cows were included in the RF group if any of the following conditions were met: the cow was in the high MY group, had low gDPR, or had at least 1 health disorder recorded. Cows were grouped into estrus groups during the VWP based on records of AEA (estrus VWP [E-VWP], n = 476 or no estrus VWP [NE-VWP], n = 410). Finally, based on the combination of levels of AEA and RF, cows were grouped into an estrus and no RF (E-NoRF, n = 217), no estrus and RF (NE-RF, n = 276), no estrus and no RF (NE-NoRF, n = 134), and estrus and RF (E-RF, n = 259) groups. Cows received AIE up to 31 d after the end of the VWP, and if they did not receive AIE, they received timed AI after an Ovsynch plus progesterone protocol. Logistic and Cox proportional hazard regression compared differences in reproductive outcomes for different grouping strategies. The NoRF (AIE: 76.9%; P/AI: 53.1%; P150: 84.5%) and E-VWP (AIE: 86.8%; P/AI: 44.8%; P150: 82.3%) groups had more cows AIE and higher P/AI and P150 than the RF (AIE: 64.5%; P/AI: 34.9%; P150: 72.9%) and NE-VWP (AIE: 50.0%; P/AI: 38.9%; P150: 72.1%) groups, respectively. When both factors were combined, the largest and most consistent differences were between the E-NoRF (AIE: 91.3%; P/AI: 58.7%; P150: 88.5%) and NE-RF groups (AIE: 47.3%; P/AI: 35.8%; P150: 69.5%). Compared with the whole population of cows or cows grouped based on a single factor, the E-NoRF and NE-RF groups had the largest and most consistent differences with the whole cow cohort. The E-NoRF and NE-RF groups also had statistically significant differences of a large magnitude when compared with the remaining cow cohort after removal of the respective group. We conclude that combining data for AEA during the VWP with other predictors of reproductive performance could be used to identify groups of cows with larger differences in expected reproductive performance than if AEA and the predictors are used alone., (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/).)

This study aimed to verify the effect of milking permission (MPE) and concentrate supplementation (CS) on milking frequency (milkings per cow per day) and milk yield (kilograms per cow per day) in a farm using a pasture-based automatic milking system (AMS). Sixty-eight cows milked using this AMS unit were randomly assigned to 1 of 4 groups homogeneous for parity, DIM, and milk yield. Treatments used were frequent or restricted MPE, that granted cows permission to milk after 6 to 8 h or 9.6 to 14 h of the previous milking, respectively; and low (LC) or high (HC) CS of 0.5 kg or 3.5 kg/cow per day, respectively. The combination of the 2 levels of MPE and the 2 levels of CS resulted in the 4 treatment combinations (frequent HC [FHC], restricted HC [RHC], frequent LC [FLC], and restricted LC [RLC]). This study was designed as a 2 × 2 factorial arrangement with treatment crossover: each of the 4 cow groups was randomly assigned to 1 of the 4 treatment combinations for a 5-wk experimental period (1 pretreatment week and 4 treatment weeks), and after each 5-wk period groups crossed over to another treatment combination until they experienced all. Statistical analysis assessed the effect of MPE, CS, and their interaction on milk yield, milking frequency, box time, milking time, and average milk-flow rate. This was done using a mixed model analysis with repeated measures to account for repeated observations on the experimental unit (cow). Milk yield per cow per day and milkings per cow per day were significantly higher with the frequent compared with the restricted MPE (1.5 kg and 0.65 milkings, respectively). Milk yield per cow per day and milkings per cow per day were significantly higher with the HC compared with the LC CS (3.1 kg and 0.25 milkings, respectively). Additionally, milk yield per cow per day was affected by the interaction of MPE and CS and it was highest with the FHC (20.1 kg) treatment combination, followed by RHC (18.2 kg) treatment combination. The number of milkings per cow per day were also affected by the interaction of MPE and CS. The highest estimated number of milkings per cow per day was recorded for the FHC (2.12) and the FLC (1.77) treatment combinations, followed by the RHC (1.38) and RLC (1.23) treatment combinations. Similarly, milking interval was 2.5 h longer for the RLC treatment combination compared with RHC. The shortest milking interval was observed for the FHC (11 h) and FLC (12.8 h) treatment combinations. In conclusion, the study showed that allowing access to the robot between 6 to 8 h after the previous milking was sufficient (even with a minimal level of CS) to achieve acceptable milk production and milking performance in a pasture-based AMS., (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/).)

Moving from conventional (CMS) to automatic (AMS) milking systems could affect milk quality. Moreover, the type and preservation methods of the forages used in the TMR, such as alfalfa hay (HTMR) or corn silage (STMR) have been demonstrated to modify milk composition. Thus, this study investigated the effect of implementing AMS and different diet forage types on the quality of Italian Holstein-Friesian bulk milk. Milk samples (n = 168) were collected monthly from 21 commercial farms in northern Italy during a period of 8 mo. Farms were categorized into 4 groups according to their milking system (CMS vs. AMS) and diet forage type (HTMR vs. STMR). Milk quality data were analyzed through the mixed procedure for repeated measurement of SAS with the milking system, diet forage type, and sampling day as fixed effects. Milking through the AMS led to lower milk fat, freezing point, and β-LG A; longer coagulation time; and higher K content, pH, and β-LG B than CMS. Cows fed STMR produced milk with greater fat, protein, casein, Mg content, titratable acidity, and β-LG A, but with reduced curd firming time, freezing point, and β-LG B than those fed HTMR. In conclusion, milk quality is not only altered by the diet's forage type and characteristics but also by the milking system., (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/).)

Our objective was to investigate the effects of intravenous (IV) or intrauterine (IU) lipopolysaccharide (LPS) challenge at 5 or 40 d postpartum (DPP) on clinical signs, systemic and uterine inflammation, dry matter intake (DMI), and milk yield (MY). Holstein cows at 5 DPP (n = 23) or at 40 DPP (n = 24) were blocked by parity and randomly assigned to 1 of 3 treatments: (1) IV-LPS (0.0625 μg/kg BW [5 DPP] or 0.1 μg/kg BW [40 DPP] over 1h), (2) IU-LPS (100 μg [5 DPP] or 300 μg [40 DPP] in 20 mL of saline), or (3) 20 mL of saline IU (IU-SAL; same for 5 and 40 DPP). The proportion of polymorphonuclear (PMN) cells was measured by endometrial cytology at d -1, 1, 4, and 7 relative to treatment. Blood haptoglobin (Hp), serum amyloid A (SAA), and LPS-binding protein (LBP), DMI, and MY were measured from d -1 through d 7. Data were analyzed separately for each DPP group in multivariable linear regression models accounting for repeated measures. Both DPP groups showed increases in rectal temperature and heart and respiratory rates, and decrease in rumination rate following IV-LPS, but not following IU-LPS. At 5 DPP, endometrial PMN proportion was similar in IU-LPS and IU-SAL. Serum Hp was unaffected by LPS challenge, SAA was greater in IV-LPS from 12 h to 24 h after challenge, and LBP was greater in IV-LPS from 8 h to 24 h. At 40 DPP, PMN was greater in IU-LPS (37 ± 4%) than in IU-SAL (15 ± 4%) 1 d after LPS challenge. Serum Hp was greater from 24 h to 72 h after challenge in IV-LPS than in the other groups, SAA was greater in IV-LPS from 6 h to 48 h, and LBP was greater in IV-LPS from 8 h to 24 h. At both 5 and 40 DPP, treatment did not affect DMI, but MY was lesser in IV-LPS cows at 12 h and 24 h than in IU-SAL or IU-LPS. The IV-LPS challenge resulted in more pronounced changes in clinical signs and acute phase protein (APP) concentrations than IU-LPS or IU-SAL at 40 DPP, but more subtle or inconsistent changes at 5 DPP. These may be due to the different doses of LPS used at 5 and 40 DPP or possibly due to the high variation in baseline clinical signs and APP observed in all groups at 5 DPP. The IU-LPS increased uterine PMN 1 d after challenge at 40 DPP but not at 5 DPP. At each time, IU-LPS did not produce changes in clinical signs or markers of systemic inflammation., (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/).)

This observational study aimed to explore the association of farmer-driven selective dry cow therapy (DCT), milking routine and dry cow management practices with SCC in early-lactation cows from 21 commercial dairy herds. Milking routine practices evaluated referred to cow preparation for milking, in-lactation mastitis management, and recording. Dry cow management practices related to dry cow environment and cleaning, dry-off procedure, milk cessation strategy and calving environment. Data from 2,016 multiparous cows in 21 commercial spring-calving grazing dairy herds were available for the study. Herd owners self-reported DCT (the assignment and administration of DCT was at the discretion of the herd owners with no involvement from the research team), management practices during milking and the dry period. Cow-level last test-day SCC records in 2020 (range = 105-285 DIM) and first test-day records in 2021 (range = 5-60 DIM) were obtained from milk recording databases. Quarter-level milk sampling was carried out on all cows in late lactation of 2020 (range = 240-261 DIM) for bacterial culturing. Bacteriological results were used to define cows with IMI when ≥1 quarter sample resulted in bacterial growth and there were no contaminated samples from that cow. Mixed model analyses were used to evaluate the association of selective DCT, milking routine, and dry cow management practices with cows' first test-day log 10 SCC (logSCC) in 2021. At dry-off in 2020, 47.6% of the cows were administered an internal teat sealant alone (ITS), while 52.4% were administered an antibiotic plus an internal teat sealant (AB+ITS). The mean herd-level percentage of cows with IMI was 19.7% (range = 9.8%-39.5%); Staphylococcus aureus accounted for the majority of cow-level IMI (89.9%, 357/397). Between herds, the proportion of cows administered ITS ranged from 17.7% (14/79; in a herd with an IMI prevalence of 10.1%) to 86.8% (66/76; in a herd with an IMI prevalence of 27.6%). In total, 11.8% (105/889) and 29.8% (292/980) of cows that were administered ITS or AB+ITS had an IMI in late lactation 2020, respectively. The mean untransformed SCC at the last test day in 2020 of cows administered ITS and AB+ITS was 55,000 and 197,200 cells/mL, respectively. The statistical analysis showed a significant interaction between selective DCT and milk yield at last test-day in 2020; cows with a milk yield of 15 kg and administered ITS had a 0.1 higher (untransformed SCC of 19,000 cells/mL higher) first test-day logSCC compared with cows administered AB+ITS. Additionally, greater parity, IMI in late lactation, higher logSCC at the last test-day in 2020 and longer dry periods were associated with higher logSCC at the first test-day in 2021. The current study identified cow- and herd-level management practices that could aid dairy farmers in improving the outcome of selective DCT and decrease early lactation SCC., (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 prospective cohort study were to characterize associations among genomic merit for fertility with ovarian and endocrine function and the estrous behavior of dairy cows during an entire nonhormonally manipulated estrous cycle. Lactating Holstein cows entering their first (n = 82) or second (n = 37) lactation had ear-notch tissue samples collected for genotyping using a commercial genomic test. Based on genomic predicted transmitting ability values for daughter pregnancy rate (gDPR), cows were classified into high (Hi-Fert; gDPR > 0.6, n = 36), medium (Med-Fert; gDPR -1.3 to 0.6, n = 45), and low fertility (Lo-Fert; gDPR < -1.3, n = 38) groups. At 33 to 39 DIM, cohorts of cows were enrolled in the Presynch-Ovsynch protocol for synchronization of ovulation and initiation of a new estrous cycle. Thereafter, the ovarian function and endocrine dynamics were monitored daily until the next ovulation by transrectal ultrasonography and concentrations of progesterone (P4), estradiol, and FSH. Estrous behavior was monitored with an ear-attached automated estrus detection system that recorded physical activity and rumination time. Overall, we observed an association between fertility group and the ovarian and hormonal phenotype of dairy cows during the estrous cycle. Cows in the Hi-Fert group had greater circulating concentrations of P4 than cows in the Lo-Fert group from d 4 to 13 after induction of ovulation and from day -3 to -1 before the onset of luteolysis. The frequency of atypical estrous cycles was 3-fold greater for cows in the Lo-Fert than the Hi-Fert group. We also observed other modest associations between genomic merit for fertility with the follicular dynamics and estrous behavior. We found several associations between milk yield and parity with ovarian, endocrine, and estrous behavior phenotypes as cows with greater milk yield and in the second lactation were more likely to have unfavorable phenotypes. These results demonstrate that differences in reproductive performance between cows of different genomic merit for fertility classified based on gDPR may be partially associated with circulating concentrations of P4, the incidence of atypical phenotypes during the estrous cycles, and, to a lesser extent, the follicular wave dynamics. The observed physiological and endocrine phenotypes might help explain part of the differences in reproductive performance between cows of superior and inferior genomic merit for fertility., (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 experiment was to investigate the effect of lipid from rapeseed cake and oats on ruminal CH 4 emission and lactational performance of dairy cows. Twelve lactating Nordic Red cows, of which 4 were primiparous, and averaging (±SD) 48 ± 22.9 DIM, 37.8 ± 7.14 kg/d milk yield were enrolled in a switch-back design experiment with 3 periods of 4 wk each. The cows were assigned into 6 pairs based on parity, DIM, milk yield, and BW at the beginning of the experiment. The experimental treatments were (1) rapeseed cake and oats (RSC+O), and (2) rapeseed meal and barley (RSM+B) as the concentrate feeds. Cows in each pair were randomly assigned to 1 of the 2 groups, which received the treatments in 2 different sequences (i.e., group 1 received RSC+O in period 1 and 3, and RSM+B in period 2, whereas group 2 was fed RSM+B in period 1 and 3, and RSC+O in period 2). The diets consisted of a partially mixed ration with grass silage mixed with either oats or barley, according to the treatment sequence, and the rapeseed cake or meal being mixed into a pellet with either oats or barley according to the treatments, and a mineral mix. The pellet was delivered at a fixed amount (i.e., 6 kg/d for multiparous and 5 kg/d for the primiparous cows) from the milking robot. The actual forage to concentrate ratios for RSC+O and RSM+B were 51:49 and 52:48, respectively, with NDF concentrations of 41.5% and 36.0% and CP concentrations of 17.0% and 16.7% of diet DM. Dry matter intake, milk yield, and gas exchange (with a GreenFeed system attached to the milking robot) were recorded daily, and milk composition and spot fecal samples were collected during the last week of each period. Based on feed analysis, and DMI of the cows during the experiment, the total fat content of the experimental diets was 4.1% and 2.7% of DM for RSC+O and RSM+B diets, respectively. Dry matter intake was 1.6 kg/d lower, and milk yield tended to be 1.0 kg/d greater for RSC+O versus RSM+B. There were no differences in ECM yield and milk composition between the treatments, whereas milk ME efficiency was greater for cows fed RSC+O than RSM+B. Methane yield (g/kg DMI) did not differ between treatments, but CH 4 production (g/d) was 9.4% and CH 4 intensity as g/kg ECM was 11.7% lower for RSC+O versus RSM+B. The lower CH 4 production was likely caused by the lower DMI and fiber digestibility, observed with the RSC+O diet. In addition, the greater lipid intake also contributed to lower rate of fermentation and subsequent decrease in CH 4 production. Overall, feeding rapeseed cake with oats in a grass silage-based diet increased feed efficiency while decreasing CH 4 emission intensity in lactating cows. This provides a practical way of mitigating ruminal CH 4 emission from dairy operations while maintaining milk production with commonly used feedstuffs in Nordic conditions., (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/).)

Key factors such as stage of lactation, parity, and body fat reserves have been associated with the digital cushion thickness (DCT), however, there are discrepancies between the results of previously published studies. The objective of this study was to examine the association of stage of lactation, body fat reserves, parity, and lesion incidence with DCT in a large cohort of intensively monitored cows. Across 4 UK farms, 2,352 cows were prospectively enrolled and assessed at 4 time points: before calving (T1-Precalving), immediately after calving (T2-Calving), in early lactation (T3-Early), and in late lactation (T4-Late). At each time point, BCS was recorded, the presence of sole lesions (sole ulcers and sole hemorrhage) and white line lesions was assessed by veterinarians, and an ultrasound image was taken to retrospectively measure the backfat thickness (BFT) in the pelvic region and the digital cushion on the hind left lateral claw. Mixed effects multivariable linear regression models, with the cow as a random effect, were fit to examine the association between the explanatory variables and DCT. The explanatory variables tested were farm, parity, stage of lactation, BCS, BFT, height, the presence of a lesion at the time of measurement, the chronicity of a lesion during early lactation, the predicted maximum daily milk yield, and the rate of milk production rise in early lactation. Stage of lactation and farm were both associated with DCT; however, an interaction was present, and this DCT pattern of change was farm-dependent. Two distinct patterns emerged; one indicated the nadir to occur shortly after calving, the other indicated the nadir to occur during early lactation. Neither BFT nor BCS were significantly associated with DCT. Heifers displayed thinner digital cushions compared with multiparous cows; however, this effect was dependent on the stage of lactation, with heifers having a thinner digital cushion up until late lactation, by which time DCT was commensurate with multiparous animals. Sole lesions and white line lesions at the time of measurement were associated with DCT (sole lesion: estimate = -0.07 mm, 95% CI = -0.14-0.00; white line lesion: estimate = 0.28 mm, 95% CI = 0.15-0.42)., (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/).)

Hypercalcemia during pregnancy is a risk for adverse maternal and fetal consequences. Although primary hyperparathyroidism is by far the most common etiology of hypercalcemia in pregnancy, an array of other etiologies of hypercalcemia associated with pregnancy and lactation have been described. Parathyroidectomy continues to be the preferred treatment for primary hyperparathyroidism. Medical management options are limited., Competing Interests: Disclosure The authors declare that they have no relevant disclosures., (Copyright © 2024 Elsevier Inc. All rights reserved.)

Information regarding protein expression and phosphorylation modifications in the bovine milk fat globule membrane is scarce, particularly throughout various lactation periods. This study employed a complete proteome and phosphoproteome between bovine colostrum and mature milk. A total of 11 proteins were seen in both protein expression and phosphorylation levels. There were 400 proteins identified in only protein expression, and 104 phosphoproteins identified in only phosphorylation levels. A total of 232 significant protein characteristics were identified within the proteome and significant phosphorylation sites within 86 phosphoproteins of the phosphoproteome. Biological activities and pathways primarily exhibited associations with the immune system. Simultaneously, a comprehensive analysis of proteins and phosphorylation sites using a multi-omics approach. Hence, the data we have obtained has the potential to expand our understanding of how the bovine milk fat globule membrane might be utilized as a beneficial component in dairy products., 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 Elsevier Ltd. All rights reserved.)

Dominance hierarchies are known for mitigating conflicts and guiding priority of access to limited resources in gregarious animals. The dominance hierarchy of dairy cows is typically investigated using agonistic interactions, often monitored at the feed bunk right after fresh feed delivery when competition is high, resulting in frequent interactions. Yet, the outcome of agonistic interactions during times of high competition may be more influenced by cows' high valuation of fresh feed than their intrinsic attributes, such that the dominance hierarchy constructed using agonistic interactions under high versus low competition times might differ. We tested how the structure of the dominance hierarchy changes in relation to different levels of competition in a dynamic group of 48 lactating dairy cows over 10 mo, with 6 cows exchanged every 16 d, for a total of 159 cows. Using a validated algorithm, we continuously detected the actor and reactor of replacement behaviors in 30 feed bins as cows competed for feed. We also calculated the percentage of occupied feed bins to characterize competition at the moment of each replacement. These data were combined to create hierarchies using Elo ratings, separately for 25 occupancy levels ranging from 13% to 100%. For each 1% rise in feeder occupancy, hierarchy steepness fell by 2.41 × 10 -3 ± 9.71 × 10 -5 (SE), and the percentage of dyads where both cows replaced each other rose by 0.13% ± 0.01%. At the highest feeder occupancy level in comparison to the lowest one, we observed 7.57% more dyads in which the dominant individual (those that won more interactions at the lowest feeder occupancy) started to lose proportionally more. The magnitude of decrease in the winning rate of the dominant individual in those dyads also got amplified by 1.06 × 10 -3 % ± 1.37 × 10 -4 % (SE) for each 1% increase in feeder occupancy. These findings illustrate how inferred hierarchies vary with competition, with high competition flattening the hierarchy due to increased success of subordinate animals. We suggest that during heightened competition, increased valuation of resources can affect competitive success more than the individual's intrinsic dominance attributes. We recommend against calculating dominance hierarchies based on agonistic interactions during periods of high competition alone, and more generally urge researchers to differentiate agonistic interactions based on context when constructing dominance hierarchies., (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/).)

Methane is a potent greenhouse gas produced during the ruminal fermentation and is associated with a loss of feed energy. Therefore, efforts to reduce methane emissions have been ongoing in the last decades. Methane production is highly influenced by factors such as the ruminal microbiome and host genetics. Previous studies have proposed to use the ruminal microbiome to reduce long-term methane emissions, as ruminal microbiome composition is a moderately heritable trait and genetic improvement accumulates over time. Lactation stage is another important factor that might influence methane production, but potential associations with the ruminal microbiome have not been evaluated previously. This study sought to examine the changes in ruminal microbiome over the lactation period of primiparous Holstein cows differing in methane intensity (MI) and estimate the heritability of the abundance of relevant microorganisms. Ruminal content samples from 349 primiparous Holstein cows with 14 to 378 DIM were collected from May 2018 to June 2019. Methane intensity of each cow was calculated as methane concentration/milk yield. Up to 64 taxonomic features (TF) from 20 phyla had a significant differential abundance between cows with low and high MI early in lactation, 16 TF during mid lactation, and none late in lactation. Taxonomical features within the Firmicutes, Proteobacteria, Melainabacteria, Cyanobacteria, Bacteroidetes, and Actinobacteria phyla were associated with low MI, whereas eukaryotic TF and those within the Euryarchaeota, Verrucomicrobia, Kiritimatiellaeota, and Lentisphaerae phyla were associated with high MI. Out of the 60 TF that were found to be differentially abundant between early and late lactation in cows with low MI, 56 TF were also significant when cows with low and high MI were compared in the first third of the lactation. In general, microbes associated with low MI were more abundant early in lactation (e.g., Acidaminococcus, Aeromonas, and Weimeria genera) and showed low to moderate heritabilities (0.03 to 0.33). These results suggest some potential to modulate the rumen microbiome composition through selective breeding for lower MI. Differences in the ruminal microbiome of cows with extreme MI levels likely result from variations in the ruminal physiology of these cows and were more noticeable early in lactation, probably due to important interactions between the host phenotype and environmental factors associated with that period. Our results suggest that the ruminal microbiome evaluated early in lactation may be more precise for MI difference, and hence, this should be considered to optimize sampling periods to establish a reference population in genomic selection scenarios., (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/).)

Introduction: Vaccination is critical in controlling the coronavirus disease 2019 (COVID-19) pandemic. However, vaccine perception and acceptance among pregnant and lactating women is unknown in Singapore. We aimed to determine the acceptance of COVID-19 vaccination among these two groups of women in Singapore and the factors associated with vaccine acceptance., Methods: We conducted an anonymous, online survey on the perceptions of the COVID-19 vaccine and its acceptance by pregnant and lactating women at a tertiary maternal and child hospital in Singapore from 1 March to 31 May 2021. Information on their demographics and knowledge was collected. These factors were assessed for their relationship with vaccine acceptance., Results: A total of 201 pregnant and 207 lactating women participated. Vaccine acceptance rates in pregnant and lactating women were 30.3% and 16.9%, respectively. Pregnant women who were unsure or unwilling to take the vaccine cited concerns about safety of the vaccine during pregnancy (92.9%), while lactating women were concerned about its potential long-term negative effects on the breastfeeding child (75.6%). Factors that were positively associated with vaccine acceptance included a lower monthly household income or education level, appropriate knowledge regarding vaccine mechanism and higher perceived maternal risk of COVID-19. Most pregnant (70.0%) and lactating women (83.7%) were willing to take the vaccine only when more safety data during pregnancy and breastfeeding were available., Conclusion: COVID-19 vaccine acceptance was low among pregnant and lactating women in Singapore. Addressing the safety concerns when more data are available and education on the mechanism of vaccine action will likely improve acceptance among these women., (Copyright © 2024 Copyright: © 2024 Singapore Medical Journal.)

Milk yield dynamics and production performance reflect how dairy cows cope with their environment. To optimize farm management, time series of individual cow milk yield have been studied in the context of precision livestock farming, and many mathematical models have been proposed to translate raw data into useful information for the stakeholders of the dairy chain. To gain better insights on the topic, this study aimed at comparing 3 recent methods that allow one to estimate individual cow potential lactation performance, using daily data recorded by the automatic milking systems of 14 dairy farms (7 Holstein, 7 Italian Simmental) from Belgium, the Netherlands, and Italy. An iterative Wood model (IW), a perturbed lactation model (PLM), and a quantile regression (QR) were compared in terms of estimated total unperturbed (i.e., expected) milk production and estimated total milk loss (relative to unperturbed yield). The IW and PLM can also be used to identify perturbations of the lactation curve and were thus compared in this regard. The outcome of this study may help a given end-user in choosing the most appropriate method according to their specific requirements. If there is a specific interest in the post-peak lactation phase, IW can be the best option. If one wants to accurately describe the perturbations of the lactation curve, PLM can be the most suitable method. If there is need for a fast and easy approach on a very large dataset, QR can be the choice. Finally, as an example of application, PLM was used to analyze the effect of cow parity, calving season, and breed on their estimated lactation performance., (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/).)

Prolonging dairy cattle longevity is regarded as one of the options to contribute to more sustainable milk production. Because failure to conceive is one of the main reasons for culling, this study investigates how adjustments in reproduction management affect partial net return at herd level and greenhouse gas emissions per unit of milk, using a dynamic stochastic simulation model. The effects of reproduction decisions that extend cattle longevity on milk yield, calving interval and pregnancy rate were derived from actual performance of Dutch commercial dairy cows over multiple lactations. The model simulated lactations, calving, and health status events of individual cows for herds of 100 cows. Scenarios evaluated differed in the maximum number of consecutive AI attempts (4, 5, or 6 services), or the production threshold (20, 15, or 10 kg of milk/d) at which cows that failed to conceive are culled (reproductive culling). Annual partial net return was computed from revenues of sold milk, calves, and slaughtered cows, and the costs from feed consumption, rearing replacement heifers, AI services, and treatment for clinical mastitis and lameness. Greenhouse gas emissions were computed for feed production, enteric fermentation, and manure management, and were expressed as total CO 2 equivalents (CO 2 -eq). Average age at culling increased with an increased maximum number of AI services. This increase was larger when going from a maximum of 4 to 5 AI attempts (108 d) than from a maximum of 5 to 6 attempts (47 d). Similarly, the average age at culling increased from 1,968 to 2,040 and 2,132 d when the threshold for reproductive culling decreased from 20, to 15 and 10 kg of milk/d, respectively. Average annual partial net return increased by 1.1% from €165,850 per 100 cows at a maximum of 4 AI to €167,670 per 100 cows at a maximum of 6 AI, and increased by 4.3% from €161,210 per 100 cows at a reproductive culling threshold of 10 kg/d to €168,190 per 100 cows at a threshold of 20 kg/d. Greenhouse gas emissions decreased by 1.2% from 0.926 to 0.915 kg CO 2 -eq per kg of fat- and protein-corrected milk (FPCM) with an increase in a maximum number of AI from 4 to 6 AI. Conversely, greenhouse gas emissions increased by 0.2% from 0.926 kg at a threshold of reproductive culling of 20 kg/d to 0.928 kg CO 2 -eq/kg FPCM at a threshold of 10 kg/d. Although lowering the threshold for reproductive culling has the potential to extend cattle longevity more than increasing the maximum number of AI services, only the increase in AI services benefits a farm's partial net return, while reducing greenhouse gas emissions., (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/).)

Our objectives were to determine the effect of oxytocin use during colostrum harvest on colostrum yield and IgG concentration in Holstein dairy cows on a commercial dairy in New York and to describe associations of cow characteristics with these outcomes. Animals were enrolled between July and October 2023 using a randomized block design, with day of enrollment as the unit of randomization. A median (range) of 10 (3-19) cows were enrolled per day. Treatments were (1) 40 IU oxytocin (OXY40), (2) 20 IU oxytocin (OXY20), and (3) an untreated control group (CNTR). Oxytocin was administered i.m. ∼45 s before unit attachment in a rotary parlor. Colostrum weight was measured using the colostrum bucket-embedded scale or a platform scale when the yield was less than the smallest bucket scale. The concentration of IgG ([IgG]) in colostrum was determined using radial immunodiffusion and used to calculate total IgG, and DM (%) was determined by oven drying. Individual cow characteristics such as parity, calf sex, weight, dystocia score, stillbirth, and milk production in wk 4 of lactation; additionally, for multiparous cows, dry period length and previous lactation dry-off linear SCS were collected. Data were analyzed separately for primiparous and multiparous cows using backward stepwise elimination. To produce final mixed effects ANOVA models, primiparous cows (n = 201) were randomized to 35.8% (n = 72) OXY40, 32.8% (n = 66) OXY20, and 31.3% (n = 63) CNTR. Multiparous groups (n = 435) were randomized to 34.7% (n = 151) OXY40, 29.7% (n = 129) OXY20, and 35.6% (n = 155) CNTR. The median (range) colostrum yield was 6.0 (0-20.6) kg and [IgG] was 98.5 (0.1-293.6) g/L in the study population. In primiparous cows, OXY40 had a higher colostrum yield (LSM [95% CI]) of 5.4 (4.9-5.9) kg compared with both OXY20 (4.1 [3.5-4.7] kg) and CNTR (3.8 [3.3-4.3] kg). In multiparous cows, OXY40, OXY20, and CNTR did not differ in colostrum yield (5.9 [5.3-6.5], 5.7 [5.1-6.3], and 5.4 [4.8-6.0] kg, respectively), but colostrum yield was greater in parity 2 compared with all other parities, cows giving birth to male calves, cows with the highest milk production at wk 4 of lactation, and cows with a dry period of >65 d. Oxytocin use did not affect [IgG] in either primiparous or multiparous cows, but [IgG] was highest in cows in parity ≥4 and lowest in cows dry >65 d. In summary, oxytocin use at 40 IU i.m. was associated with a higher colostrum yield in primiparous cows only and no change in [IgG]. Oxytocin use likely addressed disturbed milk ejection and therefore increased colostrum yield in heifers milked for the first time in a rotary parlor. This study confirms cow characteristics associated with colostrum production within a single herd., (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/).)

An experiment was conducted to identify the factors that cause reduced production of cows fed a diet with high content of corn distillers grains with solubles (DDGS). We hypothesized that the factors could be high sulfur (S) content in DDGS, which may directly (S toxicity) or indirectly (DCAD) cause reduced production. We also hypothesized that high PUFA in DDGS could be another major factor. In a randomized complete block design, 60 lactating cows (15 primiparous and 45 multiparous; average ± SD at the beginning of the trial: milk yield, 44.0 ± 6.9 kg/d; DIM, 123 ± 50; BW, 672 ± 82 kg) were blocked, and cows in each block were randomly assigned to 1 of the following treatments: SBM (4.7% fatty acids [FA], 0.22% S, and 178 mEq/kg DM of DCAD), a diet containing soybean meal as the main protein source; DG, with SBM replacing mainly soybean byproducts and supplemental fat with distillers grains at 30% dietary DM (4.7% FA, 0.44% S, and 42 mEq/kg DM of DCAD); SBM+S, SBM with sodium bisulfate for additional dietary S (4.8% FA, 0.37% S, and 198 mEq/kg DM of DCAD); SBM+CO, SBM with corn oil (4.7% FA, 0.23%, and 165 mEq/kg DM of DCAD); and DG+DCAD, DG with increased DCAD (4.7% FA, 0.40% S, and 330 mEq/kg DM of DCAD). Due to the limited number of tiestalls, blocks 1 to 6 started the experiment first as phase 1, and the rest of the blocks, as phase 2, started the experiment after phase 1. All cows were fed the SBM diet for 10 d as a covariate period followed by the experimental period for 35 d. Data were analyzed using PROC MIXED of SAS (Version 9.4, SAS Institute Inc.); block and phase were random effects; and treatments, repeated week, and interaction were fixed effects. We found an interaction of week by treatment for DMI. Although milk yield did not change, milk fat concentration tended to decrease (2.78% vs. 3.34%) for DG compared with SBM. Dry matter, OM, NDF, and CP digestibilities were lower when cows were fed the DG diet compared with SBM. Additionally, cows fed DG had lower blood concentrations of HCO 3 - , base excess, and total (t)CO 2 compared with SBM. The SBM+S diet did not affect production, nutrient digestibility, or blood parameters compared with SBM. The SBM+CO diet decreased milk fat concentration and yield compared with SBM. The DG+DCAD diet tended to increase milk fat yield and concentration (1.24 vs. 1.47 kg/d; 2.78% vs. 3.37%) and increased ECM (40.9 vs. 45.1 kg/d) compared with DG but did not improve nutrient digestibility. However, blood HCO 3 - , base excess, and tCO 2 were greater for DG+DCAD compared with DG. In conclusion, the indirect role of S - , altering DCAD, along with the high PUFA content in DDGS, appear to be the factors causing reduced production responses to a high DDGS diet. Increasing DCAD to 300 mEq/kg DM in a high DDGS diet can be a feeding strategy to alleviate reduced production responses., (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 prolactin (PRL) hormone is a major regulator of mammary gland development and lactation. However, it remains unclear whether and how PRL contributes to mammary epithelial cell proliferation and secretion. The Boer and Macheng black crossbred goats are superior in reproduction, meat, and milk, and are popular in Hubei province. To elucidate the mechanisms of PRL on mammary growth and lactation, to improve the local goat economic trade, we have performed studies on these crossbred goats during pregnancy and early lactation, and in goat mammary epithelial cells (GMECs). Here, we first found that the amino acid transporters of SNAT1 and SNAT2 expression in vivo and in vitro were closely associated with PRL levels, the proliferation and secretion of GMECs; knockdown and over-expression of SNAT1/2 demonstrated that PRL modulated the proliferation and lactation of GMECs through regulating SNAT1/2 expression. Transcriptome sequencing and qPCR assays demonstrated the effect of PRL on the transcriptional regulation of SNAT1 and SNAT2 in GMECs. Dual-luciferase reporter gene assays further verified that the binding of the potential PRL response element in the SNAT1/2 promoter regions activated SNAT1/2 transcription after PRL stimulation. Additionally, silencing of either PRLR or STAT5 nearly abolished PRL-stimulated SNAT1/2 promoter activity, suggesting PRLR-STAT5 signaling is involved in the regulation of PRL on the transcriptional activation of SNAT1/2. These results illustrated that PRL modulates the proliferation and secretion of GMECs via PRLR-STAT5-mediated regulation of the SNAT1/2 pathway. This study provides new insights into how PRL affects ruminant mammary development and lactation through regulation of amino acid transporters.

The objective of the present study was to evaluate the substitution of soybean meals by biodiesel co-products on the performance and nutrient balance of lactating cows on pasture. Twelve dairy cows (7/8 Holstein-Gir) in early lactation were included in this feeding trial. These animals had an average daily milk yield of 21.1 kg and were distributed in a 4 × 4 triple Latin square experimental design. Protein supplements fed to these lactating cows contained peanut meal, cotton and sunflower as a complete replacement for soybean meal. Nutrient intake and digestibility, milk production, microbial protein synthesis, energy and protein balance were the parameters assessed in this study. Supplements containing different biodiesel co-products did not influence (P > 0.05) the level of forage intake, supplementation and nutrient by lactating cows, except for the total digestible nutrients (TDN) intake which was higher (P ≤ 0.05) in cows supplemented with cottonseed meal. The cotton meal also promoted higher digestibility of TDN (P = 0.001) and total carbohydrates (TCHO; P = 0.001) of the diet and, as a result, higher energy consumption (P = 0.01) and lower negative energy balance (P = 0.01). Cottonseed meal is a co-product of the biodiesel industry that has the ability to replace soybean meal as a source of protein in the supplement without affecting performance and promoting greater energy availability, demonstrating its ability to reduce the effects of negative energy balance in cows at the beginning of lactation kept on tropical pastures., (© 2024. The Author(s), under exclusive licence to Springer Nature B.V.)

Accurately predicting phosphorous (P) and calcium (Ca) dietary requirements is critical for optimizing dairy cattle performance, and minimizing mineral excretions and ecosystems eutrophication. This study provides a new factorial system to determine net and dietary P and Ca requirements for maintenance and lactation, derived from a meta-regression of mineral trials involving lactating dairy cows. A comprehensive global database was constructed from 57 peer-reviewed articles of mineral balance trials, with a wide range of dietary and animal performance data. We estimated the net requirements for maintenance from the intercept of a nonlinear equation between mineral intake and the sum of total fecal and urinary excretions, which is an estimate of endogenous mineral loss. Mineral secreted in milk was used to obtain net requirements for lactation. The mineral metabolizable coefficient was quantified through observed (treatment means) mineral intake and total fecal and urinary excretions, discounting the estimated endogenous excretions from our proposed models. The nonlinear models of total fecal and urinary mineral excretion were evaluated (observed versus predicted values) using a 5-fold cross validation approach. The models to estimate the sum of endogenous fecal and urinary excretions of P (0.135±0.043 g P/kg BW0.75) and Ca (0.360±0.144 g Ca/kg BW0.75) exhibited suitable precision and accuracy; r = 0.89 and 0.79, concordance correlation coefficient = 0.85 and 0.77, and root mean square prediction error = 24.1 and 20.5% observed means, respectively. Dietary variables (forage level, fiber, starch, crude protein, and ether extract) did not affect the metabolizable coefficient (MC) of P and Ca; therefore, an overall dietary MC of P (0.69±0.01) and Ca (0.65±0.02) were proposed. Our new system estimates lower net and dietary P requirements for lactating dairy cows compared to the NASEM-2021 and NRC-2001 models, but slightly higher Ca requirements than NASEM-2021.This proposed system holds potential to reduce the use of phosphorus in diets for dairy cows, and thus to enhance economic efficiency and environmental sustainability of the dairy industry., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2024 Oliveira, Soares. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Background: Maternal breast volume is determined by the quantity of glandular and adipose tissue, and it undergoes significant changes during pregnancy. These changes are intricately linked to the development of glandular tissue, which most likely reflects lactation capacity. Evidence indicates that women with overweight or obesity exhibit larger breast volume compared to those with a normal body mass index (BMI), emphasizing the close relationship between breast volume and maternal adiposity. Hence, we aim to investigate breast volume growth and maternal BMI as potential risk factors for low milk production., Methods: Lactating women (n = 609) from the Perth metropolitan area in Western Australia between 2011 and 2023 were included in the analysis. Twenty-four-hour milk production measurements were conducted using the test weighing method, and milk removal frequencies were recorded. Mothers completed questionnaires regarding demographic, obstetric and infant details. Linear and logistic regression models were used to determine maternal and infant factors associated with milk production., Results: Here we show that increasing maternal age and BMI are associated with low milk production. Moreover, larger pre-pregnancy breast volume and breast growth are associated with both higher BMI and milk production., Conclusions: Women who are older, have an obese BMI and who have minimal pre-pregnancy breast volume and breast growth should be provided with antenatal screening and breastfeeding support as they are more likely to experience low milk production.

Postpartum breast cancer (PPBC) is a unique subset of breast cancer, accounting for nearly half of the women diagnosed during their postpartum years. Mammary gland involution is widely regarded as being a key orchestrator in the initiation and progression of PPBC due to its unique wound-healing inflammatory signature. Here, we provide dialogue suggestive that lactation may also facilitate neoplastic development as a result of sterile inflammation. Immune cells are involved in all stages of postnatal mammary development. It has been proposed that the functions of these immune cells are partially directed by mammary epithelial cells (MECs) and the cytokines they produce. This suggests that a more niche area of exploration aimed at assessing activation of innate immune pathways within MECs could provide insight into immune cell contributions to the developing mammary gland. Immune cell contribution to pubertal development and mammary gland involution has been extensively studied; however, investigations into pregnancy and lactation remain limited. During pregnancy, the mammary gland undergoes dramatic expansion to prepare for lactation. As a result, MECs are susceptible to replicative stress. During lactation, mitochondria are pushed to capacity to fulfill the high energetic demands of producing milk. This replicative and metabolic stress, if unresolved, can elicit activation of innate immune pathways within differentiating MECs. In this review, we broadly discuss postnatal mammary development and current knowledge of immune cell contribution to each developmental stage, while also emphasizing a more unique area of study that will be beneficial in the discovery of novel therapeutic biomarkers of PPBC., (© 2024. The Author(s).)

Competing Interests: None declared.

Hormonal changes in pregnant and lactating women significantly affect bone metabolism and overall stress levels, positioning them as a unique group within the orthodontic population. Fluctuations in estrogen, progesterone, prolactin, and other hormones are closely linked to bone remodeling and the periodontal tissue's response to inflammation caused by dental plaque. Hormones such as thyrotropin, leptin, and melatonin also play crucial roles in pregnancy and bone remodeling, with potential implications for orthodontic tooth movement. Additionally, adverse personal behaviors and changes in dietary habits worsen periodontal conditions and complicate periodontal maintenance during orthodontic treatment. Notably, applying orthodontic force during pregnancy and lactation may trigger stress responses in the endocrine system, altering hormone levels. However, these changes do not appear to adversely affect the mother or fetus. This review comprehensively examines the interaction between hormone levels and orthodontic tooth movement in pregnant and lactating women, offering insights to guide clinical practice., (© 2024. The Author(s).)

The main purpose was to determine the abundance of dominant phyla, Bifidobacterium spp., and Lactobacillus in breast milk of obese mothers versus normal-weights in fourth month of lactation in Iranian population. Sixty health women at the fourth month of breastfeeding, aged 18-40 years, were included and categorized based on body mass index (BMI) to the obese (BMI ≥ 30 kg/m 2 ) and normal-weights (18.5 ≤ BMI ≤ 24.9). Bacterial DNA was extracted and qPCR of the 16S region was performed after human milk donation in a sterile condition. A multiple linear mixed model was used to determine the effective factors on the phyla population. Bifidobacterium spp. was significantly higher in milk of normal-weight group than the obese. The current weight showed a significant effect on the Actinobacteria abundance in milk. The Bacteroidetes and Firmicutes were significantly lower in mother's milk with cesarean section (p = 0.04). Pre-pregnancy obesity decreased the Firmicutes and Lactobacillus abundance in maternal milk (p = 0.04 and p = 0.01). The Actinobacteria and Bifidobacterium spp. showed a significant effect on infant's height (p = 0.008 and p = 0.04). The maternal current and pre-pregnancy weight showed an important effect on abundance of Actinobacteria and Bifidobacterium spp., as the good phyla and genus in milk which are associated with the infant's height., (© 2024. The Author(s).)

COVID-19 was responsible for more than 7 million deaths globally, as well as numerous morbidities and social and economic effects. While COVID-19 vaccines were seen as a marvel of science by the scientific community, much of the public had concerns related to COVID-19 vaccines, with certain groups-such as pregnant and lactating women-having specific concerns related to vaccine effects on their pregnancy and breast milk. In this qualitative study, we interviewed stakeholders in Bangladesh ( n  = 26) and Kenya ( n  = 94) who affect the decision-making process related to COVID-19 vaccine acceptance among pregnant and lactating women. These included pregnant and lactating women themselves, community gatekeepers or family members, healthcare workers, and policymakers. Several themes related to confidence and vaccine preference emerged. Stakeholders indicated a lack of confidence related to non-mRNA vaccines due to safety concerns, number of doses, and media coverage; lack of confidence related to mRNA vaccines due to safety concerns; and preference for non-mRNA vaccines due to health system compatibility and availability. While COVID-19 vaccine availability in much of the world-particularly in low-and middle-income countries-affected the public's ability to have a choice in the vaccine they received, there were evident vaccine preferences. As the public health world will continue to face other infectious disease outbreaks, bolstering vaccine confidence broadly and specifically related to new technologies will be paramount to realize the individual-and population-level benefits of life-saving vaccines., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2024 Schue, Fesshaye, Miller, Singh and Limaye.)