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Delayed milk ejection, manifested most often as bimodal milk flow, occurs when the cisternal milk fraction is removed before the alveolar milk reaches the gland cistern. It is thought to be a consequence of not meeting cows' physiological needs, due to insufficient premilking teat stimulation, inadequate timing of milking unit attachment, or both. It has been associated with decreased milking efficiency, reduced milk yield, and impaired teat and udder health. Traditionally, portable electronic milk meters have been used to assess the presence of delayed milk ejection in dairy cows. By contrast, incremental milk flow rates from on-farm milk meters and their suitability as a measure to assess delayed milk ejection have not been studied by rigorous methods. The objectives were (1) to describe a protocol for identification of cows with chronically delayed milk ejection (CDME) and (2) to investigate risk factors for CDME using incremental milk flow rates obtained from automated on-farm milk meters. In a retrospective case control study, milk flow data from a 4,300-cow dairy with a thrice-daily milking schedule were obtained over a 1-wk period. Incremental milk flow rates (0-15 s, 15-30 s, 30-60 s, and 60-120 s) were used to identify cows with delayed milk ejection. Cases of CDME were defined as presence of delayed milk ejection at all 21 milking observations. Cows that had no delayed milk ejection at any of the same 21 milking observations were included as controls. A total of 171 cases and 393 controls were included in the study based on these criteria. A logistic regression model was used to evaluate associations of the following risk factors with CDME: parity (1, 2, ≥3), stage of lactation (<100, 101-200, >200 DIM), presence of a nonlactating quarter, milk somatic cell count, average daily milk production, and health and management events. Parity and CDME were associated such that compared with cows in their third or greater lactation, the odds (95% confidence intervals, 95% CI) of CDME were 1.27 (0.71-2.25) for cows in their first and 4.77 (2.47-9.22) for animals in their second lactation. The odds of CDME increased with increasing stage of lactation, with an odds ratio of 0.20 (0.11-0.36) for early and 0.28 (0.15-0.52) for mid-lactation animals, respectively, compared with late lactation cows. A 1-kg increase in average daily milk production was associated with decreased odds of CDME [odds ratio (95% CI): 0.89 (0.87-0.92)]. A lameness event during the study period increased the odds of CDME [odds ratio (95% CI): 8.04 (1.20-53.83)], as did a vaccination event 1 wk before the study period [odds ratio (95% CI): 4.07 (0.99-16.71)]. This study confirmed associations between CDME and previously reported risk factors and identified several previously less rigorously investigated health and management events that could be associated with CDME. Incremental milk flow rates from individual cows serve as an automated tool to evaluate milk flow dynamics. This information could be used to improve individual premilking udder preparation to meet the animal's physiological requirements, improve teat and udder health, and enhance parlor efficiency., (© 2022, The Authors. Published by Elsevier Inc. and Fass 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/).)

Antibiotic excretion into milk depends on several factors such as the compound's physicochemical properties, the animal physiology, and the milk composition. The objective of this study was to develop a physiologically based pharmacokinetic (PBPK) model describing the passage of drugs into the milk of lactating species. The udder is described as a permeability limited compartment, divided into vascular, extracellular water (EW), intracellular water (IW) and milk, which was stored in alveolar and cistern compartments. The pH and ionization in each compartment and the binding to IW components and to milk fat, casein, whey protein, calcium, and magnesium were considered. Bidirectional passive diffusion across the blood-milk barrier was implemented, based on in vitro permeability studies. The model application used to predict the distribution of oxytetracycline in cow and goat milk, after different doses and routes of administration, was successful. By integrating inter-individual variability and uncertainty, the model also allowed a suitable estimation of the withdrawal periods. Further work is in progress to evaluate the predictive ability of the PBPK model for compounds with different physico-chemical properties that are potentially actively transported in order to extrapolate the excretion of xenobiotics in milk of various animal species including humans., (Copyright © 2022 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Serotonin plays a diverse role in maternal and mammary metabolism. Recent research in the dairy cow has shown a relationship between serotonin and calcium, with increased serotonin concentrations improving calcium homeostasis in the peri-partum dairy cow. Therefore, the objective was to elucidate how administration of 5-hydroxy-l-tryptophan (5-HTP), the immediate precursor to serotonin, altered serotonin and calcium metabolism in lactating dairy cows. Twelve mid-late lactation multiparous cows were blocked by parity, production and days in milk and allocated to a daily intravenous infusion of (i) 1.5 mg/kg of 5-HTP (n = 6) or (ii) saline (n = 6) for 3 consecutive days. Milk samples were collected daily. Blood samples were collected before and after each infusion with mammary biopsies and blood samples collected at 48, 56, and 72 h relative to termination of first infusion. Infusion of 5-HTP increased (p = 0.001) circulating serotonin concentrations and decreased blood calcium via a transient hypocalcemia immediately after each infusion (p = 0.02). Treatment with 5-HTP increased milk calcium concentrations (p = 0.02) and calcium release-activated channel protein 1 (ORAI1) mRNA at 56 h and protein at 48 h relative to termination of first infusion (p = 0.008 and p = 0.09, respectively). Fifty-six hours from termination of the first infusion mRNA of parathyroid hormone-related protein and mammary serotonin content were increased relative to control (p = 0.03 and p = 0.05, respectively). These findings demonstrate the ability of 5-HTP infusion to increase circulating serotonin concentrations and alter endocrine and mammary autocrine/paracrine calcium and serotonin metabolism in the lactating dairy cow., (© 2021 The Authors. Physiological Reports published by Wiley Periodicals LLC on behalf of The Physiological Society and the American Physiological Society.)

Mastitis remains the most expensive disease of dairy cows, and antibiotic dry cow therapy (DCT) at dry-off is an important part of mastitis control. Regardless of the infection status, blanket DCT is administered to all quarters of all cows, which is controversial due to the worldwide problem of antimicrobial resistance. Even though selective DCT of only infected cows is a more sustainable approach, choosing animals for treatment is not always straightforward. Our aim was to evaluate whether the herd-level DCT approach is associated with early lactation udder health problems, taking into account the cow characteristics. The information source was 2015-2017 Dairy Herd Improvement data with 7461 multiparous cows from 241 Finnish dairy herds. Information on the herd-level DCT approach was obtained from farmers' questionnaire responses in 2017, and the three different approaches were selective DCT, blanket DCT, and no DCT. The statistical tool for the data analysis was a generalized linear mixed model with a random herd effect for binary outcomes and a linear mixed model with a random herd effect for a continuous outcome. The two binary outcomes were the odds of having high milk somatic cell count (SCC ≥ 200 000 cells/mL) on the first test-day within 5-45 days in milk (DIM) and the odds of mastitis treatment in early lactation up to 45 DIM. The third outcome was the mean milk lnSCC (× 1000 cells/mL) within 120 DIM. Selective DCT was the prevailing treatment practice in our data. Blanket DCT was associated with lower SCC after calving. Cows more likely to have high SCC after calving were older cows, cows with high average SCC during the previous lactation, and cows with high milk yield near dry-off. A mastitis treatment in the early lactation was more likely if, during the previous lactation, the cow had high average SCC, high peak milk production, or high milk yield near dry-off. Our findings indicate that DCT is still effective in mastitis control. Cows with high milk yield, especially near dry-off, and cows with persistently high SCC require attention when considering next lactation udder health., (Copyright © 2021 The Author(s). Published by Elsevier B.V. All rights reserved.)

Shortening or omitting the dry period to improve the energy balance in early lactation have the trade-offs of reduction in milk production and loss of opportunity for dry-cow therapy (DCT; i.e., intramammary antibiotic use at dry-off). Customized dry-period strategies (i.e., deciding upon DCT and dry-period length per cow) could mitigate negative effects of short or no dry periods on milk production and udder health and simultaneously retain benefits from improved energy balance and fertility. In this study, we evaluated 3 decision trees to customize dry-period strategies for individual cows. In the control tree (CT), all cows had a 60-d dry period, with DCT if somatic cell count (SCC) was >150,000 cells/mL before dry-off. In decision tree 1 (T1), parity 1 and parity >1 cows were assigned DCT if SCC was ≥150,000 cells/mL and SCC ≥50,000 cells/mL, respectively; whereas in decision tree 2 (T2), the threshold for DCT was SCC ≥200,000 cells/mL for all animals. In T1 and T2, cows with DCT were assigned a 60-d dry period, whereas cows without DCT were assigned a 30-d or 0-d dry period if their milk production remained >12 kg/d at 67 and 37 d before calving, respectively. Cows were monitored from 8 wk before to 14 wk after calving. Milk production and composition, SCC, body condition score, body weight, and occurrence of treatment for disease (related to calving and start of lactation) were compared between CT (n = 61 cows), T1 (n = 59 cows), and T2 (n = 63 cows). Effects of decision trees (CT, T1, T2) and of dry-period strategies (60-d dry with or without antibiotics, 30-d dry, or 0-d dry) on measured variables were analyzed separately with mixed models, effects on udder-health status with a logistic regression, and occurrence of treatment for diseases with a Pearson chi-squared test. In T1, 36% of cows qualified for 30-d and 2% for 0-d dry periods, whereas in T2 this was 51% and 30% for 30-d and 0-d dry periods, respectively. Compared with CT, cows in T1 and T2 on average produced more milk in the 8 wk before calving (0.2 vs. 3.9 vs. 7.1 kg/d in CT vs. T1 vs. T2), and less in the 14 wk after calving (40.0 vs. 37.0 vs. 35.2 kg/d in CT vs. T1 vs. T2). There was no difference in udder-health status in the transition period among decision trees. In the first 14 wk after calving, recovery of body weight was greater for T2 than CT and T1. Overall, 30-d and 0-d dry periods reduced milk revenues, but this might be financially compensated by improved cow health with customized dry-period strategies., (The Authors. Published by Elsevier Inc. and Fass Inc. on behalf of the American Dairy Science Association®. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).)

Resilience is the ability of cows to cope with disturbances, such as pathogens or heat waves. To breed for improved resilience, it is important to know whether resilience genetically changes throughout life. Therefore, the aim was to perform a genetic analysis on 2 resilience indicators based on data from 3 periods of the first lactation (d 11-110, 111-210, and 211-340) and the first 3 full lactations, and to estimate genetic correlations with health traits. The resilience indicators were the natural log-transformed variance (LnVar) and lag-1 autocorrelation (r auto ) of daily deviations in milk yield from an expected lactation curve. Low LnVar and r auto indicate low variability in daily milk yield and quick recovery, and were expected to indicate good resilience. Data of 200,084 first, 155,784 second, and 89,990 third lactations were used. Heritabilities were similar based on different lactation periods (0.12-0.15 for LnVar, 0.05-0.06 for r auto ). However, the heritabilities of the resilience indicators based on full first lactation were higher than those based on lactation periods (0.20 for LnVar, 0.08 for r auto ), due to lower residual variances. Heritabilities decreased from 0.20 in full lactation 1 to 0.19 in full lactation 3 for LnVar and from 0.08 to 0.06 for r auto . For LnVar, as well as for r auto , the strongest genetic correlation between lactation periods was between period 2 and 3 (0.97 for LnVar, 0.96 for r auto ) and the weakest between period 1 and 3 (0.81 for LnVar, 0.65 for r auto ). Similarly, for both traits the genetic correlation between full lactations was strongest between lactations 2 and 3 (0.99 for LnVar, 0.95 for r auto ) and weakest between lactations 1 and 3 (0.91 for LnVar, 0.71 for r auto ). For LnVar, genetic correlations with resilience-related traits, such as udder health, ketosis, and longevity, adjusted for correlations with milk yield, were almost always favorable (-0.59 to 0.02). In most cases these genetic correlations were stronger based on full lactations than on lactation periods. Genetic correlations were similar across full lactations, but the correlation with udder health increased substantially from -0.31 in lactation 1 to -0.51 in lactation 3. For r auto , genetic correlations with resilience-related traits were always favorable in lactation period 1 and in most full lactations, but not in the other lactation periods. However, correlations were weak (-0.27 to 0.15). Therefore, as a resilience indicator for breeding, LnVar is preferred over r auto . A multitrait index based on estimated breeding values for LnVar in lactations 1, 2, and 3 is recommended to improve resilience throughout the lifetime of a cow., (The Authors. Published by Elsevier Inc. and Fass Inc. on behalf of the American Dairy Science Association®. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).)

Liner overpressure is a quantitative variable indicating the extent to which the vacuum difference across the liner during phase d (the liner compression phase) of milking machine pulsation exceeds the vacuum difference that would be just sufficient to stop milk flow from the teat. Previously defined methods of determining liner overpressure have required modifications to the milking machine, complex instrumentation, or both. Our method of measuring derived overpressure (OP) offers relatively simple instrumentation and realistic milking machine characteristics. We determined derived OP by measuring the duration of milk flow within a pulsation cycle, and then comparing that duration with the shape of the pulsation curve to deduce the pulsation chamber vacuum level corresponding to that duration. Derived OP by our method yielded measurements of OP that differed by less than 2.0 kPa from those determined by the most practical previous method, for 2 trial liners. Derived OP can serve as a method for comparing and evaluating liners, and the method we developed may also be applied to automatic control of the milking process., (Copyright © 2021 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

During the last decade, the use of systematic crossbreeding in dairy cattle herds has increased in several countries of the world. The aim of this study was to estimate the effect of breed proportion and heterosis on milk production traits and udder health traits in dairy cattle. The study was based on records on milk yield (MY), protein yield (PY), fat yield (FY), somatic cell score (SCS), and mastitis (MAST) from 73,695 first-lactation dairy cows in 130 Danish herds applying systematic crossbreeding programs. Around 45% of the cows were crosses between Danish Holstein (DH), Danish Red (DR), or Danish Jersey (DJ), and the remaining were purebred DH, DR, or DJ. The statistical model included the fixed effects of herd-year, calving month, and calving age and an effect representing the lactation status of the cow. In addition, the model included a regression on calving interval from first to second lactation, a regression on the proportion of DH, DR, and DJ genes, and a regression on the degree of heterozygosity between DH and DR, DH and DJ, and DR and DJ. Random effects were the genetic effect of the cow and a residual. The effect of breed proportions was estimated relatively to DH. For MY, a pure DR yielded 461 kg milk less than DH, whereas a pure DJ yielded 2,259 kg milk less than a pure DH. Compared with DH, PY was 41.7 kg less for DJ, whereas PY for DR was 4.0 kg less than for DH. For FY, a DR yielded 10.6 kg less than DH, whereas there was no significant effect of breed proportion between DJ and DH. A DR cow had lower SCS (0.13) than DH, whereas DJ had higher SCS (0.14) than DH. There was no significant effect of breed proportion on MAST between the 3 breeds. Heterosis was significant in all combinations of breeds for MY, FY, and PY. Heterosis for crosses between DH and DR was 257 kg (3.2%), 11.9 kg (3.2%), and 8.9 kg (3.2%) for MY, PY, and FY, respectively. Corresponding figures for crosses between DH and DJ were 314 kg (4.4%), 14.3 kg (4.4%), and 10.4 kg (4.0%), whereas heterosis between DR and DJ was 462 kg (6.7%), 19.6 kg (6.7%), and 13.9 kg (5.4%) for MY, PY, and FY, respectively. Heterosis was only significant for SCS in the crosses between DH and DR. Heterosis effects for MAST were nonsignificant for all the crosses. The results obtained in this study demonstrate that in first lactation cows, there is a positive effect of heterosis on milk production traits, but limited effect on udder health traits., (The Authors. Published by Elsevier Inc. and Fass Inc. on behalf of the American Dairy Science Association®. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).)

The objectives were to study the effect of 2 different premilking stimulation regimens, with and without manual forestripping, on teat tissue condition and milking characteristics in dairy cows. In a randomized controlled crossover study, 130 Holstein cows milked 3 times daily were assigned to treatment and control groups. Premilking udder preparation for the treatment group consisted of: (1) predipping with 1% iodine, (2) sequential forestripping of 3 streams of milk per quarter, (3) wiping of teats, and (4) attachment of the milking unit. Premilking udder preparation for the control group was identical except that the forestripping step was omitted. The mean tactile stimulation durations were 16 s and 7 s for the treatment and control group, respectively. The time spent from first tactile stimulus (either forestripping or wiping of teats) to milking unit attachment was kept consistent at 90 s for both groups. The study lasted for 14 d with 2 periods, each consisting of a 2-d adjustment time followed by 5 d of data collection. Machine milking-induced short-term changes to the teat tissue were assessed by palpation and visually. The following milking characteristics were assessed with electronic on-farm milk meters: milk yield (MY), milking unit-on time (MUOT), 2-min MY (2MIN), and time spent in low milk flow rate (LMF). Generalized linear mixed models were used to describe the effect of treatment on the outcome variables. The odds of machine milking-induced short-term changes to the teat tissue were lower for cows that received forestripping compared with cows that were not forestripped (odds ratio = 0.31; 95% confidence interval = 0.22-0.42). Least squares means (95% confidence interval) for cows that were forestripped and animals that were not forestripped, respectively, were 12.7 (12.2-13.2) and 12.7 (12.2-13.2) kg for MY and 6.1 (5.8-6.4) and 5.6 (5.3-5.9) kg for 2MIN. There was an interaction between treatment and MY for LMF. Time spent in LMF for cows that were forestripped and received no forestripping, respectively, were 18 (17-20) and 24 (23-26) s for a MY level of 10 kg; and 13 (12-14) and 15 (14-16) s for a MY level of 15 kg. The effect of treatment on MUOT was modified by parity. Milking unit-on times for animals in first, second and third or greater lactation, respectively, were 230 (219-243), 249 (236-262), and 260 (249-272) s for cows that were forestripped, and 245 (232-258), 252 (239-266), and 268 (257-281) s for cows that received no forestripping. In this study, cows that were forestripped had shorter MUOT, higher 2MIN, lower LMF, and lower odds of exhibiting changes to the teat tissue after machine milking. We conclude that wiping of teats during premilking udder preparation alone and omitting forestripping of teats without compensating for the loss in stimulation time may not provide sufficient tactile stimulation to elicit the cows' maximum physiological milk-ejection capacity. This can aggravate the adverse effects of vacuum-induced forces on teat tissue during machine milking, diminish animal well-being, and possibly affect udder health., (Copyright © 2020 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

Background: Over the last years, genome-wide association studies (GWAS) based on imputed whole-genome sequences (WGS) have been used to detect quantitative trait loci (QTL) and highlight candidate genes for important traits. However, in general this approach does not allow to validate the effects of candidate mutations or determine if they are truly causative for the trait(s) in question. To address these questions, we applied a two-step, within-breed GWAS approach on 15 traits (5 linked with milk production, 2 with udder health, and 8 with udder morphology) in Montbéliarde (MON), Normande (NOR), and Holstein (HOL) cattle. We detected the most-promising candidate variants (CV) using imputed WGS of 2515 MON, 2203 NOR, and 6321 HOL bulls, and validated their effects in three younger populations of 23,926 MON, 9400 NOR, and 51,977 HOL cows., Results: Bull sequence-based GWAS detected 84 QTL: 13, 10, and 30 for milk production traits; 3, 0, and 2 for somatic cell score (SCS); and 8, 2 and 16 for udder morphology traits, in MON, NOR, and HOL respectively. Five genomic regions with effects on milk production traits were shared among the three breeds whereas six (2 for production and 4 for udder morphology and health traits) had effects in two breeds. In 80 of these QTL, 855 CV were highlighted based on the significance of their effects and functional annotation. The subsequent GWAS on MON, NOR, and HOL cows validated 8, 9, and 23 QTL for production traits; 0, 0, and 1 for SCS; and 4, 1, and 8 for udder morphology traits, respectively. In 47 of the 54 confirmed QTL, the CV identified in bulls had more significant effects than single nucleotide polymorphisms (SNPs) from the standard 50K chip. The best CV for each validated QTL was located in a gene that was functionally related to production (36 QTL) or udder (9 QTL) traits., Conclusions: Using this two-step GWAS approach, we identified and validated 54 QTL that included CV mostly located within functional candidate genes and explained up to 6.3% (udder traits) and 37% (production traits) of the genetic variance of economically important dairy traits. These CV are now included in the chip used to evaluate French dairy cattle and can be integrated into routine genomic evaluation.

Recent advances on milk exosomes (EXO), cargoes in cell-cell communication, explored their role within and between individuals, including in dairy species. The potential use of EXO as biomarkers of disease and metabolic conditions adds significant interest to the study of EXO in milk. Although several researches have been carried out on circulating miRNA in the milk, less information is available about milk-derived exosomal miRNAs, which are stable over time and resistant to digestion and milk processing. EXO are taken up by recipient cells through specific mechanisms, which enable the selective delivery of cargoes. This suggests that EXO cargoes can be used as biomarkers of health. Nevertheless, methodological limitations and potential applications of milk EXO in dairy ruminants must be considered. The paucity of studies that associate the EXO cargo to specific challenges deserves further investigations to unravel the variation of miRNA and proteins cargo in relation to metabolic imbalance and infectious disease of the mammary gland.

Records of methane emissions from 1,501 cows on 14 commercial farms in 4 regions of Spain were collected from May 2018 to June 2019. Methane concentrations (MeC) were measured using a nondispersive infrared methane detector installed within the feed bin of the automatic milking system during 14- to 21-d periods. Rumination time (RT; min/d) was collected using collars with a tag that registered time (minutes) spent eating and ruminating. The means of MeC and methane production (MeP) were 1,254.28 ppm and 182.49 g/d, respectively; mean RT was 473.38 min/d. Variance components for MeC, MeP, and RT were estimated with REML using pedigree and genomic information in a single-step model. Heritabilities for MeC and MeP were 0.11 and 0.12, respectively. Rumination time showed a slightly larger heritability estimate (0.17). The genetic correlation between MeP and MeC was high (>0.95), suggesting that selection on either trait would lead to a positive correlated response on the other. Negative correlations were estimated between RT and MeC (-0.24 ± 0.38) and MeP (-0.43 ± 0.35). Methane concentration and MeP had slightly positive correlations with milk yield (0.17 ± 0.39 and 0.21 ± 0.36), protein percentage (0.08 ± 0.32 and 0.30 ± 0.45), protein yield (0.22 ± 0.41 and 0.31 ± 0.35), fat percentage (0.02 ± 0.40 and 0.27 ± 0.36), and fat yield (0.27 ± 0.28 and 0.29 ± 0.28) from bivariate analyses. Rumination time had positive correlations with milk yield (0.41 ± 0.75) and protein yield (0.26 ± 0.57) and negative correlations with fat yield (-0.45 ± 0.32), protein percentage (-0.15 ± 0.38), and fat percentage (-0.40 ± 0.47). A positive approximated genetic correlation was estimated between fertility and MeC (0.10 ± 0.05) and MeP (0.18 ± 0.05), resulting in slightly higher CH 4 production when selecting for better fertility [days open estimated breeding values (EBV) are expressed with mean 100 and SD 10, inversely related to days from calving to conception; that is, greater days open EBV implies better fertility]. Positive correlations were also estimated for stature with MeC and MeP (0.30 ± 0.04 and 0.43 ± 0.04, respectively). Other type traits (chest width, udder depth, angularity, and capacity) were positively correlated with methane traits, possibly because of higher milk yield and higher feed intake from these animals. Rumination time showed positive EBV correlations with production traits and type traits, and negative correlations with somatic cell count and body condition score. Based on the genetic correlations and heritabilities estimated in this study, methane is measurable and heritable, and estimates of genetic correlations suggest no strong opposition to current breeding objectives in Spanish Holsteins., (Copyright © 2020 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

The objective of this study was to compare the reproductive performance, milk yield and composition, and udder health and conformation traits of Holstein (Ho), F1, and R1 crossbred Ho × Simmental (Sim) cows. Three commercial dairy farms in south Brazil were used as the research units. All farms held Ho, F1, and R1 crossbred Ho × Sim (¾ Ho × ¼ Sim and ¾ Sim × ¼ Ho) cows. The collection of milk samples and evaluation of udder conformation traits occurred during four visits to each farm. In addition to the actively collected data, retrospective reproduction records of the farms served as the basis for the statistical analysis using analysis of variance models using SAS. The F1 crossbred Ho × Sim cows and ¾ Sim (first rotational crossbreeding generation = R1 using Sim semen) cows had a shorter calving interval and calving to first service interval compared to the Ho cows (P < 0.0001). Milk yield did not differ among the genetic groups except for R1 (¾ Sim) that produced approximately 10% less milk than the other groups (P = 0.0245). Fat plus protein yield and somatic cell score did not differ among the genetic groups. Ho cows had shallower udders (P < 0.0001) and a higher udder clearance (P < 0.0001) than the other groups. F1 and R1 crossbred Ho × Sim cows had shorter reproduction intervals than purebred Ho cows. Although udder conformation traits lacked high-quality scores in crossbred cows, somatic cell scores reached the same level as in purebred Ho cows.

To find out the effect of reducing energy intake during dry period on milk production, udder health, and body condition, the experiment was conducted on 14 Jersey crossbred cows during whole dry period and continued up to 120 days of lactation. Reduction in energy intake was done during far-off period for each dry cow of treatment group as compared to control group. Statistically analyzed data revealed that overall significantly (P < 0.01) lower DMI and WI were recorded in control than treatment group. Overall significantly (P < 0.01) higher total milk production was found in treatment than control group. Overall significantly (P < 0.01) lower milk SCC, MCMT, pH, and EC were found in treatment than control group. Nonsignificant difference in milk fat, SNF, total solid, total protein, and fat:protein ratio was found. Overall significantly (P < 0.01) better quality milk (MBRT) was found in treatment than control groups. BCS during dry period and at calving was significantly (P < 0.01) different between groups. Significantly (P < 0.01) higher plasma NEFA concentration was estimated in control than treatment groups in all stages. No significant difference was found for plasma concentrations of glucose, urea, and total protein. The coefficients of correlation indicated significant (P < 0.01) correlation among BCS, milk production, milk SCC, MCMT, pH, and EC. It can be concluded that reducing energy intake during far-off dry period can lead to achieve optimum BCS at calving. Suitable BCS at calving was beneficial to get higher milk production with improved quality, better maintenance of udder health and body condition of Jersey crossbred cows at tropical lower Gangetic region.

This research communication describes the influence of diet, mammary quarter position and milking process on the temperature of teats and udder of cows fed diets containing different lipid sources. Five primiparous cows were fed diets containing cottonseed, sunflower seed, soybeans or soybean oil as a source of lipids and a reference diet without the inclusion of lipid sources in a 5 × 5 Latin Square design. Milk yield was determined in the last five days of each period. Milk samples were collected for SCC analysis on the last two days of each experimental period. The images of the mammary gland were obtained using an infrared camera and were analyzed with appropriate computer software. Milk yield was 14.8% higher for cows fed soybeans as a source of lipids. Diets and somatic cell counts did not influence the temperature of teats and udder. The milking process reduced the temperature of teats and udder by 0.79°C. Rear teats and rear quarters had higher surface temperatures than front teats and fore quarters. Changes in temperature of teats and mammary quarters occurred as a function of the milking process and quarter position. However, the diet and the SCC did not influence the temperature of teats and mammary quarters in this experiment.

The objectives were to study the effect of 2 different automatic cluster remover settings on (1) milking characteristics, (2) milk component yields, (3) teat tissue condition, and (4) udder health. In a randomized controlled field trial, Holstein cows (n = 689) from 1 commercial dairy farm with a thrice-daily milking schedule were allocated to 2 treatment groups. Treatment consisted of a cluster remover take-off milk flow threshold of 1.2 (ACR1.2) or 0.8 kg/min (ACR0.8) for 57 d. Milking characteristics (milk yield; and milking unit-on time, MUOT) were obtained with electronic on-farm milk meters. Composite milk samples were collected and analyzed for fat, protein, lactose, and somatic cell count. Machine-milking-induced short- and long-term changes to the teat tissue condition were assessed visually. General linear mixed models demonstrated differences in MUOT, whereas no meaningful differences in milk yield were detected. Milk yield (least squares means, 95% confidence interval) was 11.3 (10.9-11.8) and 11.3 (10.8-11.8) kg in groups ACR1.2 and ACR0.8, respectively. The effect of treatment on MUOT was modified by parity. Milking unit-on time in first-, second-, and ≥third-lactation cows, respectively, was 260.7 (252.0-269.4), 257.8 (247.4-268.1), and 260.2 (252.6-267.9) s in group ACR1.2; and 273.7 (264.9-282.5), 279.1 (269.4-288.8), and 295.7 (287.9-303.6) s in group ACR0.8. We detected no meaningful differences in milk component yields or linear somatic cell score. Least squares means in groups ACR1.2 and ACR0.8, respectively, were milk fat yield, 0.42 (0.40-0.44) and 0.42 (0.40-0.44) kg; milk protein yield, 0.36 (0.35-0.37) and 0.37 (0.36-0.37) kg; milk lactose yield, 0.61 (0.60-0.63) and 0.63 (0.61-0.64) kg, and linear somatic cell score, 1.9 (1.8-2.0) and 1.9 (1.8-2.0). A generalized linear mixed model revealed an effect of treatment on machine-milking-induced short-term changes. The odds of short-term changes to the teat tissue were lower for cows in group ACR1.2 [odds ratio (95% confidence interval) = 0.78 (0.63-0.96)]. No meaningful differences were detected in machine-milking-induced long-term changes between treatment groups. Increasing cluster remover take-off milk flow threshold from 0.8 to 1.2 kg/min decreased individual milking duration and alleviated machine-milking-induced short-term changes to the teat tissue without adversely affecting milking performance or somatic cell count. Future studies are warranted to investigate the effect on milk production and udder health over a whole lactation period., (Copyright © 2020 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

The mechanisms that dairy cattle respond to environmental stresses are very complicated. Previous research into the molecular mechanisms of mammalian heat stress has largely focused on the role of protein-coding genes and small non-coding RNAs. Recently, it has become apparent that large numbers of long non-coding RNAs transcribed from mammalian genomes play extensive roles in transcriptional regulation. However, the expression of lncRNAs and their biological functions in heat stress in dairy cattle remain unknown. In this study, we employed a deep RNA sequencing to examine lncRNA expression profiles of heat stressed and non-heat stressed Chinese Holstein cattle. We found that 24,795 novel and 3763 known lncRNAs were expressed in the bovine mammary gland, of which 174 were differentially expressed in heat stress condition, among them, 156 lncRNAs were up-regulated and 18 were down-regulated. Through Cis role analysis, 16,474 lncRNAs were transcribed close to protein-coding neighbors. In addition, 11 and 2024 lncRNAs harbored precursors of known and predicted microRNAs, respectively, were annotated in the precursor analysis of miRNAs. Taken together, our findings represent the first systematic investigation of lncRNA expression in heat stressed Chinese Holstein and provide a resource for further research into the molecular mechanisms of lncRNAs function in dairy cattle.

Eleven mid-lactation Holstein cows were milked twice daily during the first 2 experimental weeks. During wk 3 to 10, the cows were differentially milked: right quarters were milked thrice daily (3×) and left quarters were milked once daily (1×). During wk 11 to 14, all quarters were milked twice daily. After 4 wk of differential milking, the cows received daily i.m. injections of the dopamine antagonist domperidone (DOMP; 300 mg; n = 6) or of dimethyl sulfoxide as the control (CTL; n = 5) for 8 wk (wk 7-14). During the differential milking period (wk 3-6), milk production was greater for quarters milked 3× than for those milked 1× but did not differ between DOMP and CTL cows. During the differential milking + injection period (wk 7-10), milk production continued to differ according to milking frequency. However, DOMP injection did not have an effect or interact with milking frequency on milk production. During the injection period (wk 11-14), milk production remained greater in the quarters previously milked 3× and milk production increased in DOMP injected cows but not in CTL cows. Injections of DOMP increased prolactin concentration, which was greater in the serum of DOMP cows than in that of CTL cows during the differential milking + injection and the injection periods. The expression of genes that are directly related to milk synthesis (CSN2, LALBA, and ACACA) was greater in the 3× quarters than in the 1× quarters. In addition, DOMP increased CSN2 expression during the injection period. The expression of both isoforms of the PRLR gene was greater in the 3× quarters during the differential milking + injection and the injection periods. At the protein level, injections of DOMP tended to increase the number of long PRLR isoform during the differential milking + injection period. The number of short PRLR isoform was greater in the 1× quarters than in the 3× quarters during the differential milking, the differential milking + injection, and the injection periods. The total amount of STAT3 protein was greater in the 1× quarters during the differential milking and the differential milking + injection periods. The amount of phosphorylated STAT3 protein was greater in the 1× quarters during the differential milking period. The total amount of phosphorylated STAT5 protein was greater in the 3× quarters during the differential milking and the differential milking + injection periods. The results of this experiment support the hypothesis that the responsiveness of the mammary gland to PRL is modulated by milking frequency, although the underlying mechanism remains to be determined., (Crown Copyright © 2020. Published by FASS Inc. and Elsevier Inc. on behalf of the American Dairy Science Association®. All rights reserved.)

The success in competitions may be stressful for animals and costly in terms of immune functions and longevity. Focusing on Aosta Chestnut and Aosta Black Pied cattle, selected for their fighting ability in traditional competitions, this study investigated the genetic relationships of fighting ability with udder health traits (somatic cell score and two threshold traits for somatic cells), longevity (length of productive life and number of calvings) and test-day milk, fat and protein yield. Herdbook information and phenotypic records that have been routinely collected for breeding programs in 16 years were used for the abovementioned traits. Data belonged to 9328 cows and 19 283 animals in pedigree. Single-trait animal model analyses were run using a Gibbs sampling algorithm to estimate the variance components of traits, and bivariate analyses were then performed to estimate the genetic correlations. Moderate positive genetic correlations (ra) were found for fighting ability with somatic cell score (ra=0.255), suggesting that greater fighting ability is genetically related to a detriment in udder health, in agreement with the theory. The high positive genetic correlation between fighting ability and longevity (average ra=0.669) suggests that the economic importance of fighting ability (the winning cows get an higher price at selling) had probably masked the true genetic covariances. The genetic correlation between milk yield traits and fighting ability showed large intervals, but the negative values (average ra=-0.121) agreed with previous research. This study is one of the few empirical studies on genetic correlations for the competitive success v. immune functions and longevity traits. The knowledge of the genetic correlations among productive and functional traits of interest, including fighting ability, is important in animal breeding for a sustainable genetic improvement.

Fully automated on-line analysis equipment is available for analysis of somatic cell count (SCC) at every milking in automatic milking systems. In addition to results from on-line cell counters (OCC), an array of additional cow-level and quarter-level factors considered important for udder health are recorded in these systems. However, the amount of variability in SCC that can be explained by available data is unknown, and so is the proportion of the variability that may be due to physiological or normal variability. Our aim was to increase our knowledge on OCC as an indicator for disturbances in udder health by assessing the variability in OCC in cows free from clinical mastitis. The first objective was to evaluate how much of the variability in OCC could be explained by different potential sources of variability, including intramammary infection (IMI) status (assessed by bacterial culture of quarter milk samples). The second objective was to evaluate the repeatability of the OCC sensor used in our study and the agreement between OCC values and SCC measured in a dairy herd improvement (DHI) laboratory. A longitudinal study was performed in the research herd of the Norwegian University of Life Sciences from January 5th 2016 to May 22nd 2017. Data from 62,471 milkings from 173 lactations in 129 cows were analyzed. We used ln-transformed OCC values (in 1000 cells/ml) as the outcome (lnOCC) in linear mixed models, with random intercepts at cow-level and lactation-level within cow. We were able to explain 15.0% of the variability in lnOCC with the following fixed effects: lactation stage, parity, milk yield, OCC in residual milk from the previous milking, inter-quarter difference between the highest and lowest conductivity, season, IMI status, and genetic lineage. When including the random intercepts, the degree of explanation was 55.2%. The individual variables explained only a small part of the total variability in lnOCC. We concluded that physiological or normal variability is probably responsible for a large part of the overall variability in OCC in cows without clinical mastitis. This is important to consider when using OCC data for research purposes or in decision-support tools. Sensor repeatability was evaluated by analyzing milk from the same sample multiple times. The coefficient of variation was 0.11 at an OCC level relevant for detection of subclinical mastitis. The agreement study showed a concordance correlation coefficient of 0.82 when comparing results from the OCC with results from a DHI laboratory., (Copyright © 2019 The Authors. Published by Elsevier B.V. All rights reserved.)

Despite their heritability and influence on female productivity, there are currently no genetic evaluations for teat and udder structure in Canadian Angus cattle. The objective of this study was to develop optimal genetic evaluations for these traits in the Canadian Angus population. Guidelines recommended by Beef Improvement Federation (BIF) were used to score teat and udder structure in 1,735 Canadian Angus cows from 10 representative herds. Cows scored ranged in parity from 1 to 13; however, >70% of cows were parity ≤4. Scores ranged from 1 (large, bottle shaped) to 9 (very small) for teats and from 1 (very pendulous) to 9 (very tight) for udders. Consistent with parity distribution, >70% of teat and udder scores were ≥6. Teat and udder scores (TS9 and US9, respectively) were modeled using a multiple trait animal model with random effects of contemporary group (herd-year-season) and additive genetic effect, and fixed effects of breed, parity group, and days between calving and scoring. To test good versus poor structure, a binary classification of 1 or 2 (TS2, US2) [comprised of scores 1 to 5 = 1 (poor structure) and scores 6 to 9 = 2 (good structure)] was created. Further, to assess the impact of grouping less frequently observed poor scores, a 1 to 7 scale (TS7, US7) was created by combining teat and udder scores 1 to 3. Analyses for teat and udder scores on scales TS9, US9, TS7, US7, and TS2, US2 were compared. In addition, both threshold and linear animal models were used to estimate variance components for the traits. Data treatment and models were evaluated based on correlation of resulting estimated breeding value (EBV) with corrected phenotypes, Spearman's rank correlation coefficient, average EBV accuracies (r), and deviance information criteria (DIC). TS9, US9 scales for teat and udder scores and linear models performed best. Estimates of heritability (SE) for teat and udder score were 0.32 (0.06) and 0.15 (0.04), respectively, indicating these traits were moderately heritable and that genetic improvement for teat and udder scores was possible. Estimates of phenotypic and genotypic correlations for teat and udder score were 0.46 (0.02) and 0.71 (0.09), respectively. Estimates of genotypic correlations with birth weight (BW), weaning weight (WW), and yearling weight (YW), ranged from -0.04 (0.10) to -0.20 (0.12), verifying the importance of selecting for improved teat and udder score as individual traits, alongside performance traits., (© The Author(s) 2019. Published by Oxford University Press on behalf of the American Society of Animal Science.)

Bovine mammary gland involution, as a part of the reproductive cycle in dairy cows, is a very important remodeling transformation of the mammary gland for the subsequent lactation. There is considerable incentive to accelerate mammary gland involution to improve udder health, shorten the dry period, and simplify the management process by reducing dietary changes. The complex process of mammary involution is characterized by morphological changes in the epithelial cells and mammary tissue, changes in the composition of mammary secretions, and changes in the integrity of tight junctions. Involution is facilitated by elements of the immune system and several types of proteases and is coordinated by various types of hormones. This review first describes the involution process and then argues for the need to accelerate it. Last, this review focuses on various intervention methods for accelerating involution. Our aim is to provide a comprehensive overview of bovine mammary gland involution as well as potential techniques and new opinions for dry cow management., (Copyright © 2019 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

Shortening or omitting the dry period improves the energy balance and metabolic status of dairy cows in early lactation. Metabolic, behaviour and welfare effects throughout lactation, however, are unclear. The current paper reviews long-term metabolic and welfare consequences of short and no dry period, as well as feeding strategies and individual cow characteristics that could support in optimising management of cows with a short or no dry period. The paper will conclude with impacts of short and no dry periods at herd level and in practice. Energy balance after no or a short dry period is more positive during the complete subsequent lactation. After the initial improvement in early lactation, cows after no dry period tend to fatten and may have a too low lactation persistency to be continuously milked until the onset of the subsequent lactation. Reducing dietary energy level for cows with no dry period reduced fattening during the complete lactation but did not improve lactation persistency. Feeding a more lipogenic diet for cows with a short or no dry period did not affect the energy balance or lactation persistency during the complete lactation, although a lipogenic diet resulted in lower plasma insulin and IGF-1 concentration and greater plasma growth hormone concentration, compared with a glucogenic diet. Effects of dry period length on udder health are ambiguous, whereas short and no dry periods improved fertility in most studies. Omission of the dry period changed behaviour of cows both before and after calving, with a longer lying time and greater feed intake after calving, suggesting a better adaptation to a new lactation. Individual cow characteristics like parity, genotype, prepartum body condition score, and milk yield level determined the metabolic response of cows to a short or no dry period. In conclusion, short or no dry periods increase the energy balance in the complete lactation. Feeding strategies can be used to limit fattening of cows with no or short dry period, but the studied feeding strategies did not increase lactation persistency. Improved fertility and behavioural changes around calving suggest a better adaptation to a new lactation in case of no dry period. Customised dry period lengths for individual cows could improve metabolic status of cows at risk of a severe negative energy balance while minimising milk losses.

Because of technical limitations, an impact of machine milking on the teat tissue cannot be avoided. The continuance of this impact during and after milking depends on a variety of factors related to the physiological regulation of milk ejection, as well as the different production systems and milking machine settings. Milking machine settings aim to achieve a high milking performance, that is, short machine-on time at a maximum of milk harvest. However, a high milking performance level is often related to an impact on the teat tissue caused by vacuum or liner compression that can lead to pathological dimensions of congestion of the tissue or hyperkeratosis as a long-term effect. Toward the end of milking a decrease of milk flow rate causes a raise of mouthpiece and teat end vacuum levels and hence an increase of the impact on the teat tissue and the risk of tissue damage. The mechanical stress by the milking machine activates a cascade of cellular mechanisms that lead to an excessive keratin growth and thickening of the keratin layer. Consequently, a complete closure of the teat canal is disabled and the risk of bacterial invasion and intramammary infection increases. Another consequence of high vacuum impact is fluid accumulation and congestion in the tissue of teat tip and teat basis because of an obstruction in venous return. The present review paper provides an overview of the available scientific information to describe the interaction between different levels and types of system vacuum, mouthpiece chamber vacuum, teat end (claw) vacuum, liner pressure, and the risk of short-term and long-term impacts on the teat tissue.

Milking cows once daily is a management tool that has been implemented to improve physical and financial results of seasonal pasture-based dairy farms. The Molly cow model integrates physiology and metabolism of dairy cattle; however, milk production during short-term changes in milking frequency (e.g., 1× milking) is not well represented. The model includes a representation of variable rates of cell quiescence and death. However, the rate constants governing cell death and the return of quiescent to active cells are not affected by milking frequency. An empirical assessment of the problem was conducted, and it was hypothesized that changing the current representation of the rate of cell death in response to short-term 1× milking would more accurately represent active and quiescent cells and improve predictions of milk production. An extra senescent cell flux was added to account for cell loss during periods of 1× milking. Additional changes included a gradual decline in the rate of 1× stimulated senescence during 1× milking, and a structural change in cell cycling between active and quiescent cells during and after short-term 1× milking. Data used for parameter estimation were obtained from 5 studies where 1× milking or different feeding strategies were tested. Parameter estimates of cell loss indicated that 1× milking would affect a small proportion of quiescent cells to cause extra cell death. This added cell senescence was influenced by the length of 1× milking such that cell senescence peaked on d 1 of 1× milking and decayed from that point. The new structure in the model includes a variable rate of cell death in response to 1× milking and a gradual rate of return of quiescent cells back to the active pool in response to switching to 2× milking after short-term 1× milking. Root mean square errors, mean bias, and slope bias declined by at least 50% for predictions of energy-corrected milk yield and fat percent. The model showed quantitative agreement with production data from short-term 1× milking. The accuracy of predictions was improved and the error was reduced by implementing modifications in the model in response to changes in milking frequency., (Copyright © 2019 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

Accumulating evidence supports that the hormone prolactin (PRL) is galactopoietic in dairy ruminants. Accordingly, the inhibition of PRL secretion by the dopamine agonists quinagolide and cabergoline causes a sharp decline in milk production and could be useful in several critical periods. First, PRL inhibition may reduce the incidence during the periparturient period of metabolic disorders caused by the abrupt increase in energy demand for milk production. Metabolic disturbances can be lessened by reducing milk output by milking once a day or incompletely in the first few days of lactation. The injection of cows with quinagolide for the first 4 days of lactation reduced milk production during the first week of lactation without any residual effects. Blood glucose and calcium concentrations were higher and β-hydroxybutyric acid concentration was lower in the quinagolide-treated cows. Second, PRL inhibition may help sick or injured lactating cows, considering that they can fall into severe negative energy balance when they are unable to consume enough feed to support their milk production. This leads to a weakened immune system and increased susceptibility to diseases. When cows were subjected to feed restriction and were treated with quinagolide, the decrease in milk production was accelerated without any residual effects. The quinagolide-treated cows had higher glucose and lower β-hydroxybutyric acid and non-esterified fatty acid concentrations than the control cows did. Third, PRL inhibition may facilitate drying-off in high-yielding cows, because they are often dried off while still producing significant quantities of milk, which delays mammary involution and increases risk of mastitis. Therefore, strategies that reduce milk production before drying-off and accelerate mammary gland involution could be an important management tool. In this context, inhibition of PRL was utilised to accelerate mammary gland dry-off. Quinagolide decreased milk production within the first day of treatment, and both quinagolide and cabergoline induced more rapid changes in several markers of mammary gland involution after drying-off. In addition, quinagolide improved the animals' resistance to intramammary infection. These results suggest that the inhibition of PRL could be a strategy for facilitating drying-off, reducing metabolic stress during the postpartum period, and alleviating acute nutritional stress during illness without compromising the overall productivity of dairy ruminants.

Historically, pre-pubertal development of the bovine mammary gland (MG) has received little attention compared to later development. Recent evidence suggests not only that this period represents a very active time in the development of the MG but also that the first 90 days of life can partially dictate future productivity of the lactating cow. The MG, often considered quiescent during early life (first 3 months), is now known to increase in size by over 60-fold in the same period. The importance of sex steroids in MG development is well classified, but a complex signaling network exists among estrogen, progesterone and other growth factors and hormones. Complicating our understanding of this developmental period further is the discovery that pre-weaning nutrition of the calf not only influences the growth of the mammary parenchyma but may also alter the way in which it responds to mammogenic stimuli. Recent data suggest that feeding calves a higher plane of nutrition improves the ability of the mammary epithelium to respond to estradiol and also alters the way in which the mammary parenchyma and fat pad communicate. It is clear that early life nutrition, although able to influence the MG, is still poorly understood mechanistically. For example, additional evidence suggests that increased feeding rates in early life alter the morphology of myoepithelial cells in the mammary epithelium. Further data have also suggested a role for other cell types, such as immune cells, in the penetration of the mammary parenchyma into the fat pad during the early life development of the MG suggesting that mammary development is not only controlled by the local tissue population (parenchyma and fat pad) but perhaps systemically by other tissue types (i.e., immune system). Understanding the roles of these various stimuli and signaling pathways as they relate to the development of the MG in early life may hold the key to unlocking the potential for the optimal development of this crucial organ and, in turn, may lead to improvements in other phases of mammary development and milk yield potential.

An extended milking interval of 24 h (24-h milking interval (24h-MI)) constitutes the acute phase of cow adaptation to once-daily milking (ODM). A recent trial including 724 24h-MI challenges demonstrated that milk yield responses to this acute phase of ODM are highly variable (from+22% to -52% of milk yield when switching to the 24h-MI, mean=-25.3%) and that factors such as stage of lactation parity and milk yield level influenced cows' responses but did not account for all individual variability. Additional traits related to physiological, immune and behavioural adaptation were measured on a subset (96 observations) of this data set. This study aimed to determine (1) the relationship of these traits with cows' milk yield responses, (2) their ability - combined with previously identified traits - to help predict milk yield responses to 24h-MI (adaptive profiles). The 24h-MI challenge consisted of three successive periods: one control week of twice-daily milking (cTDM), one single day of 24h-MI and then 13 days of TDM (pTDM). Milk yield responses to the 24h-MI (corrected for effects of stage of lactation, parity, milk yield level and milk yield) were related to physiological traits measured during cTDM (milk flow rate, presence or absence of interleukin-8) and to their changes during the 24h-MI (absolute increase in milk flow rate and relative udder distension). Analysis of associations between milk yield responses, stage of lactation, parity, milk yield level, proteolysis, udder expansion and immune traits found three adaptive cow profile clusters. Cows in cluster 1 had a less compliant udder than cows in cluster 2, and they lost more milk during the 24h-MI than cluster-2 and cluster-3 cows. After resuming twice daily-milking (TDM), cluster-2 cows fully recovered the milk they had lost during the 24h-MI. On the opposite, cluster-3 cows did not recover the milk they lost, likely due to udder inflammation during cTDM, as suggested by elevated concentrations of interleukin-8 in their milk. These results combining new traits with stage of lactation, parity and milk yield level constitute a first step towards predicting individual cow responses to a 24h-MI.

Increased milking frequency and incomplete milking have differential effects on milk yield and mammary gland physiology that are important for optimization of milking practices in dairy herds. The objectives of this experiment were to determine the effects of increased milking frequency and incomplete milking on milk production rate (MPR) and milk composition and to determine if milking 3 times daily (3×) could rescue the negative production effects of incomplete milking. Twenty-two multiparous cows were enrolled onto this experiment beginning at 5 days in milk (DIM) and continuing through 47 DIM. A split-plot design was used to randomize the 2 treatments, which were milking frequency and incomplete milking. Eleven cows were randomly assigned to be milked 2 times (2×) daily and 11 cows were randomly assigned to be milked 3×. Within each cow, a contralateral half-udder was randomly assigned to be incompletely milked (30% milk remaining in the gland; IM), and the other half-udder was randomly assigned to be milked completely (CM). Quarter-level milk yields were recorded at each milking session. Milk samples from all quarters were collected twice weekly at the beginning of the morning milking for analysis. Cows milked 2× tended to have reduced MPR compared with 3× milked cows (1.81 ± 0.06 vs. 1.97 ± 0.06 kg milk/h; P = 0.06). Half-udders that were CM and IM produced 1.09 ± 0.03 and 0.80 ± 0.03 kg milk/h, respectively. There was an interaction between incomplete milking treatment and week of lactation (P = 0.04). No interaction was detected between milking frequency and incomplete milking for MPR or milk components. Cows milked 3× had increased milk fat percent (1.93 ± 0.09% vs. 1.65 ± 0.09%, P = 0.047), decreased milk lactose percent (4.80 ± 0.04% vs. 4.93 ± 0.04%, P = 0.04), and exhibited no differences in milk protein percent or milk somatic cell count (SCC) compared with cows milked 2×. Half-udders that were IM had increased milk fat percent (2.15 ± 0.07% vs. 1.43 ± 0.07%, P < 0.0001), decreased lactose percent (4.75 ± 0.03% vs. 4.99 ± 0.03%, P < 0.0001), increased milk log10SCC (4.22 ± 0.05 vs. 4.41 ± 0.05, P = 0.0004), and no differences in milk protein percent compared with CM half-udders. These results indicate that a 3× milking frequency in IM half-udders was not able to improve milk production compared with IM half-udders milked 2×. Our results indicate that 30% milk remaining in the gland had an irreversible impact on milk yield as increased milking frequency was not able to reverse the milk yield lost., (© The Author(s) 2019. Published by Oxford University Press on behalf of the American Society of Animal Science. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

The objective of this study was to assess the effect of treating cows with teat sealant only compared with antibiotic plus teat sealant at drying off on weekly somatic cell count, potential intramammary infection, and milk production across the entire subsequent lactation. In 3 research herds in the south of Ireland, cows with SCC that did not exceed 200,000 cells/mL in the previous lactation (LowSCC) were randomly assigned to 1 of 2 treatments at drying off: internal teat sealant alone (ITS) or antibiotic plus teat sealant (AB+ITS). Cows with SCC that exceeded 200,000 cells/mL in the previous lactation were treated with AB+ITS and included in the analyses as a separate group (HighSCC). Weekly individual animal composite SCC records were available for 654 cow lactations and were transformed to somatic cell scores (SCS) for the purpose of analysis. Data were divided into 3 data sets to represent records obtained (1) up to 35 DIM, (2) up to 120 DIM, and (3) across the lactation. Foremilk secretions were taken from all quarters at drying off, at calving, 2 wk after calving, and in mid-lactation and were cultured to detect the presence of bacteria. The LowSCC cows treated with ITS alone had higher daily milk yield (0.67 kg/d) across lactation compared with LowSCC cows treated with AB+ITS. The LowSCC cows treated with ITS alone had higher SCS in early, up to mid, and across lactation compared with LowSCC cows treated with AB+ITS. We detected no difference in weekly SCS of LowSCC cows treated with ITS alone and SCS of HighSCC cows. The least squares means back-transformed SCC across lactation of the LowSCC cows treated with ITS alone, LowSCC cows treated with AB+ITS, and HighSCC cows were 41,523, 34,001, and 38,939 cells/mL respectively. The odds of LowSCC cows treated with ITS alone having bacteria present in their foremilk across lactation was 2.7 (95% confidence interval: 1.91 to 3.85) and 1.6 (1.22 to 2.03) times the odds of LowSCC cows treated with AB+ITS and of HighSCC cows treated with AB+ITS, respectively. In this study, Staphylococcus aureus was the most prevalent pathogen isolated from the population. Recategorizing the threshold for LowSCC cows as ≤150,000 cells/mL or ≤100,000 cells/mL in the previous lactation had no effect on the results. The results indicate that herds with good mastitis control programs may use ITS alone at dry-off in cows with SCC <200,000 cells/mL across lactation with only a small effect on herd SCC., (Copyright © 2019 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

Automatic cluster remover (ACR) settings regulate the end of milking by detaching the clusters based on milk flow dropping below a preset level, which needs to be standardised for different breeds of dairy animals based on their production. A study was conducted to find out the best ACR setting for milking Indian crossbred cows based on milkability, milking irregularities and milk quality. Fifty six crossbred dairy cows in lactations 1 to 4 were categorised into three groups based on the level of production; low (N = 16; 18 kg/d). The ACR settings tested were 0.1, 0.2, 0.3 and 0.4 kg/min, keeping the vacuum level and pulsation settings constant. The ACR settings significantly (P &lt; 0.01) affected the milk yield at all levels of production with a significant effect (P &lt; 0.01) on machine-on time at 0.4 kg/min. The yield during the first 2 min of milking, average flow and peak flow rates were not affected at any level of production. The average electrical conductivity in milk was significantly (P &lt; 0.01) lower for the low and medium yield cows without affecting the mean somatic cell count. At 0.4 kg/min, more cluster reattachments were needed because of significant amount of milk remaining in the udders post-cluster removal.

Milk production of dairy cows has increased markedly during recent decades and continues to increase further. The evolutionarily conserved direction of nutrients to the mammary gland immediately after calving provided the basis for successful selective breeding toward higher performance. Considerable variation in adaptive responses toward energy and nutrient shortages exists; however, this variation in adaptability recently gained interest for identifying more metabolically robust dairy cows. Metabolic challenges during periods of high milk production considerably affect the immune system, reproductive performance, and product quality as well as animal welfare. Moreover, growing consumer concerns need to be taken into consideration because the public perception of industrialized dairy cow farming, the high dependency on feed sources suitable for human nutrition, and the apparently abundant use of antibiotics may affect the sales of dairy products. Breeding for high yield continues, but the metabolic challenges increasingly come close to the adaptational limits of meeting the mammary gland's requirements. The aim of the present review is to elucidate metabolic challenges and adaptational limitations at different functional stages of the mammary gland in dairy cows. From the challenges and adaptational limitations, we derive perspectives for sustainable milk production. Based on previous research, we highlight the importance of metabolic plasticity in adaptation mechanisms at different functional stages of the mammary gland. Metabolic adaptation and plasticity change among developing, nonlactating, remodeling, and lactational stages of the mammary gland. A higher metabolic plasticity in early-lactating dairy cows could be indicative of resilience, and a high performance level without an extraordinary occurrence of health disorders can be achieved., (Copyright © 2019 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

The objective of this study was to determine the herd-level variables that were associated with total stimulation time during the premilking routine in 64 Michigan dairy herds. The mean herd size was 452 cows (range = 59 to 2,771 cows). For each herd, surveys were administered to producers to gather mastitis management practices and attitudes. Additionally, milking protocols were observed and milk flow dynamics were measured by use of digital vacuum recorders. Backward multivariate regression analysis was used to determine which of 47 herd-level milking and management variables were associated with mean total stimulation time. Mean total stimulation time was 14.2 s (range = 2.4-40.8 s) and was positively associated with increasing latency period (time interval between first stimulation and cluster attachment). Total stimulation time was negatively associated with greater herd size and number of visits to each cow in the premilking routine. In summary, increased stimulation time is more likely in herds that foster a lower sense of urgency of cow throughput during milking, as evidenced by a positive association with longer latency periods and fewer preparation visits per cow. Tactile stimulation is critical for efficient milk ejection; if inadequate, cows are at greater risk of delayed milk ejection and bimodal milk flow, which in turn has been associated with teat congestion and reduced milk flow. This study offers insight as to some of the herd factors that may be limiting adequate tactile stimulation., (Copyright © 2019 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

Successful lactation in multiparous dairy cattle relies on a well-managed dry period that allows the mammary gland to remodel and regenerate between lactations. Oxylipids are potent inflammatory mediators that are capable of regulating all aspects of inflammation. Although an oxylipid profile has been documented for periparturient and lactating cattle, little work has been done to define the profile of cows in the early dry period. Therefore, our group aimed to characterize the oxylipid profile in healthy cows during the transition into early mammary gland involution. Plasma samples were collected from 10 healthy Holstein dairy cows via coccygeal venipuncture 6 d before dry-off (d -6), at dry-off (d 0), and 1 (d +1), 2 (d +2), 6 (d +6), and 12 (d +12) d after the dry-off date. Liquid chromatography-mass spectrometry was used to quantify select monounsaturated fatty acids, polyunsaturated fatty acids, and saturated fatty acids, whereas oxylipids were quantified using liquid chromatography-tandem mass spectrometry. The results of this study revealed a unique profile of pro- and anti-inflammatory oxylipids throughout the transition from late lactation into the dry period. Many compounds reached the highest concentrations of the study at d +1, d +2, or d +12, whereas others reached the lowest concentrations at d +12. The characterization of this profile allows for further understanding of the physiology of early mammary involution. Future studies should investigate how the oxylipid profile of early mammary involution may affect the health and productivity of dairy cows., (Copyright © 2019 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

Background: Premilking udder preparation is essential for harvesting high-quality milk as gently, completely, and quickly as possible. The associations between characteristics such as teat-end shape and premilking stimulation on milking characteristics and machine milking-induced changes to the teat tissue condition have not been rigorously investigated. The primary objective was to investigate the interactive effects of manual premilking stimulation (i.e., preparation lag time) and teat-end shape on total milk yield, two-minute milk yield, milking unit-on time, and time in low milk flow rate. Our secondary objective was to study the association of manual premilking stimulation and changes to the teat tissue condition after machine milking (i.e., short-term changes). In a longitudinal prospective cohort study, 384 milking observations from 129 cows were analysed. Holstein cows were housed in sand-bedded free-stall pens, fed a total mixed ration, and milked 3 times a day. Cows were classified by teat-end shape into 1 of 3 categories: pointed, flat, or round. Individual cow milking characteristics were recorded with electronic on-farm milk meters. The duration of manual stimulation, preparation lag time, and presence of short-term changes were documented for each milking observation. General linear mixed models were used to study the interactive effects of preparation lag time and teat-end shape on milking characteristics., Results: There was an interaction between preparation lag time and teat-end shape for two-minute milk yield and time in low milk flow rate. The preparation lag time effect was modified by teat-end shape, while no interaction was observed for total milk yield or milking unit-on time. A generalized linear mixed model revealed that preparation lag time was associated with short-term changes in teat tissue condition, where the odds of short-term changes decreased as preparation lag time increased., Conclusions: In summary, cows with different teat-end shapes may require different premilking stimulation regimens. Increasing preparation lag time benefits teat tissue condition during machine milking. Further research is warranted to optimize individual premilking stimulation in dairy cows.

The objective of this study was to establish different single or multiple trait selection indices to calculate genetic and economic gains by combining some production, reproduction and udder health traits in a population similar to the overall practical situation in Iran, with and without imposing restrictions on genetic change for some traits. The SelAction software was used to perform the analyses based on selection index theory through a deterministic model. Results indicated that among established indices, the index that showed the highest genetic gain for milk yield did not maximize the total genetic and economic gains. Rather, the index that included all production, reproduction and udder health traits yielded the highest genetic and economic gains. When we placed restriction on the selection indices, the economic gain decreased and the amount of reduction depended on the heritability and the correlation of restricted trait(s) with other traits. Generally, regarding the economic genetic gain per generation, the indices based on records of 200 offspring were 4.819% more efficient than those that used information of 100 offspring.

Crossbreeding in dairy cattle has recently become of increased interest. However, farmers in Scandinavian countries are reluctant to implement crossbreeding in their herds, and one reason is the common opinion that only herds at a poor level of management can benefit from crossbreeding. The Danish Cattle Database (SEGES, Aarhus, Denmark) provided data on 14 traits regarding milk yield, udder health, fertility traits, stillbirth, and survival. The data were collected from 103,307 pure Holstein cows and 14,832 F 1 crosses (Holstein dam and Nordic Red sire). The cows were born between 2008 and 2014 and originated from 424 herds that contributed data from at least 5 purebreds and 5 crossbreds across the years. We split the animals into 3 production levels: high, average, and low according to the herd's average production (kg) of 305-d fat plus protein in the given birth year of the cow. We estimated least squares means of breed group (purebred and crossbred) performance within each production level. Crossbred performance in 305-d fat yield in first-parity cows was greater than that of Holstein across all herd production levels; the gain was greater in high- (9 kg more than Holstein) and average-producing herds (7 kg more than Holstein) than in low-producing herds (3 kg more than Holstein). Regardless of production level or parity, crossbreds did not outperform Holstein in terms of 305-d protein yield (0 to 8 kg less). Crossbreds had relatively better udder health than Holstein in both first and second parity (up to 15% less mastitis) within any of the production levels. In terms of fertility, stillbirth, and survival, crossbreds performed better than purebreds, and improved performance was independent of herd production level. We conclude that differences in performance between F 1 crossbreds and Holstein are independent of production level., (The Authors. Published by FASS Inc. and Elsevier Inc. on behalf of the American Dairy Science Association®. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).)

Shortening the dry period improves postpartum energy balance, which has potential positive effects on metabolic health. This concept has been mainly studied in Holstein cows. The aim of this study was to assess the effect of a short dry period of 4 wk, compared with a standard dry period of 8 wk, on the metabolic status, progesterone profiles, health, and colostrum quality of dairy cows of 2 breeds, Swedish Red (SR) and Swedish Holstein (SH), not treated with antibiotics at dry off. The IgG uptake in calves was also studied to reflect the colostrum quality when shortening the dry period. Cows of both SH and SR were blocked by breed and parity and then randomly allocated to a short dry period of 4 wk (4W, n = 43) or a conventional dry period of 8 wk (8W, n = 34). Blood samples were collected wk -8, -4, -2, -1 and 1, 2, 3, 4, 6, 9, and 12 relative to calving. Prepartum, cows with a 4-wk dry period had higher concentrations of nonesterified fatty acids and lower concentrations of insulin-like growth factor-1 and insulin than 8W cows. Postpartum, plasma concentration of nonesterified fatty acids was lower, whereas plasma insulin and insulin-like growth factor-1 tended to be higher for 4W cows than for 8W cows. Plasma concentration of β-hydroxybutyrate did not differ between dry period lengths. Swedish Holstein cows with a 4W dry period responded with a lower concentration of insulin prepartum than SR and SH on an 8W dry period. The dry period length had no effect on the proportion of disturbed progesterone profiles; disturbed progesterone profiles occurred in 30% of the 4W cows and 47% of the 8W cows. In this trial, only 48.8% of the SR cows had a normal progesterone profile, which differed from the SH where 76.5% had a normal profile. Fertility-related diseases (endometritis, pyometra, anestrus, ovarian cyst) did not differ between the 2 dry period groups: 21% in the 8W group versus 12% in the 4W group, whereas mastitis tended to be more common: 26% of the 4W cows versus 9% of the 8W cows. A short dry period resulted in less colostrum but with a higher content of protein and somatic cell count. Calves were fed colostrum from their dam, and the IgG and total protein in plasma did not differ between calves to mothers with different a dry period length. Shortening the dry period could improve metabolic status in cows of both SH and SR breed postpartum, without compromising the colostrum quality. Health and progesterone profiles were not affected by the dry period length for SH or SR in this study., (Copyright © 2019 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

Udder health problems are often associated with milk losses. These losses are different between quarters, as infected quarters are affected both by systemic and pathogen-specific local effects, whereas noninfected quarters are only subject to systemic effects. To gain insight in these losses and the milk yield dynamics during disease, it is essential to have a reliable reference for quarter-level milk yield in an unperturbed state, mimicking its potential yield. We developed a novel methodology to predict this quarter milk yield per milking session, using an historical data set of 504 lactations collected on a test farm by an automated milking system from DeLaval (Tumba, Sweden). Using a linear mixed model framework in which covariates associated with the linearized Wood model and the milking interval are included, we were able to describe quarter-level yield per milking session with a proportional error below 10%. Applying this model enables us to predict the milk yield of individual quarters 1 to 50 d ahead with a mean prediction error ranging between 8 and 20%, depending on the amount of historical data available to estimate the random effect covariates for the predicted lactation. The developed methodology was illustrated using 2 examples for which quarter-level milk losses are calculated during clinical mastitis. These showed that the quarter-level mixed model allows us to gain insight in quarter lactation dynamics and enables to calculate milk losses in different situations., (Copyright © 2018 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

Branched-chain volatile fatty acids (BCVFA) supplements could promote lactation performance and milk quality by improving ruminal fermentation and milk fatty acid synthesis. This study was conducted to evaluate the effects of BCVFA supplementation on milk performance, ruminal fermentation, nutrient digestibility and mRNA expression of genes related to fatty acid synthesis in mammary gland of dairy cows. A total of 36 multiparous Chinese Holstein cows averaging 606±4.7 kg of BW, 65±5.2 day in milk (DIM) with daily milk production of 30.6±0.72 kg were assigned to one of four groups blocked by lactation number, milk yield and DIM. The treatments were control, low-BCVFA (LBCVFA), medium-BCVFA (MBCVFA) and high-BCVFA (HBCVFA) with 0, 30, 60 and 90 g BCVFA per cow per day, respectively. Experimental periods were 105 days with 15 days of adaptation and 90 days of data collection. Dry matter (DM) intake tended to increase, but BW changes were similar among treatments. Yields of actual milk, 4% fat corrected milk, milk fat and true protein linearly increased, but feed conversion ratio (FCR) linearly decreased with increasing BCVFA supplementation. Milk fat content linearly increased, but true protein content tended to increase. Contents of C4:0, C6:0, C8:0, C10:0, C12:0, C14:0 and C15:0 fatty acids in milk fat linearly increased, whereas other fatty acids were not affected with increasing BCVFA supplementation. Ruminal pH, ammonia N concentration and propionate molar proportion linearly decreased, but total VFA production and molar proportions of acetate and butyrate linearly increased with increasing BCVFA supplementation. Consequently, acetate to propionate ratios linearly increased. Digestibilities of DM, organic matter, CP, NDF and ADF also linearly increased. In addition, mRNA expressions of peroxisome proliferator-activated receptor γ, sterol regulatory element-binding factor 1 and fatty acid-binding protein 3 linearly increased, mRNA expressions of acetyl-coenzyme A carboxylase-α, fatty acid synthase and stearoyl-CoA desaturase quadratically increased. However, lipoprotein lipase mRNA expression was not affected by treatments. The results indicated that lactation performance and milk fat synthesis increased with BCVFA supplementation by improving ruminal fermentation, nutrient digestibility and mRNA expressions of genes related to milk fat synthesis.

The objective of the present study was to quantify the relationship between udder skin surface temperature (USST) and somatic cell count (SCC) in lactating dairy cows. Data were recorded on the same 14 Holstein-Friesian cows, at evening (15:00 to 16:00) milking every day over a 2-mo period. Surface temperature measurements of all udders were extracted from thermal images. After imaging, milk was extracted from each quarter and analyzed for SCC. Environmental and cow-related factors (i.e., ambient temperature, humidity, rainfall, wind speed, distance walked to the parlor, number of days since the udder was shaved, parity, and stage of lactation) were recorded on each day of the experiment. A large array of descriptive temperature parameters (DTP) were extracted from every udder image including temperature-based (e.g., maximum, average and minimum USST), pixel count-based, and textural-based DTPs. Several different analytical methods were tested in an attempt to relate any given DTP to SCC; this included investigating the relationship between USST and the log transform of SCC (i.e., somatic cell score; SCS). The temperature range within each udder was also compared with the natural log of the range in SCC of the respective quarters. In a separate analysis, the temperature difference between each DTP and its respective daily baseline (i.e., average of the 5 lowest values of that DTP across the herd) was compared with SCS. Finally, the association between environmental and cow-related factors with each DTP was investigated to create prediction models for each DTP, the residuals of which were compared with SCC. Results from the present study indicate that the correlation between any DTP and SCS was weak (range of -0.16 to 0.19) and so could not be used to identify quarters with high SCC. Although some alternative measures had a significant relationship with SCS, again, the correlation was too weak for practical use on its own. Maximum and average USST could be predicted with a root mean square error of 0.23 and 0.35 °C, respectively, although the residuals from the prediction model could not be used to identify animals with high SCC. This suggests that infrared thermography alone could not be used as a real-time automated tool to detect high SCC for dairy cows in a pasture-based system.

Yak milk is superior to common cow milk in nutrients including protein, fat and calories. However, the milk yield of the yak is very much lower compared with other dairy bovines. To understand the molecular mechanisms of lactogenesis, lactation and mammary gland development, mammary tissue samples were taken from five yaks during a dry period (DP, n = 3) and lactation period (LP, n = 2). Two types of cDNA sequence libraries that reflected the different physiological states of the mammary gland were constructed using RNA sequencing technology. After removing reads containing adapters, reads containing poly-N and low-quality reads from the raw data, 45,423,478 to 53,274,976 clean reads were obtained from these libraries. A total of 74.72% to 80.65% of the high-quality sequence reads were uniquely aligned to the BosGru v2.0 yak reference genome. Using the DESeq R package, 360 differentially expressed genes were detected between the two groups when the adjusted P value (padj < 0.05) was used as the cutoff value; this included 192 upregulated and 168 downregulated genes in the yak mammary gland tissue of the DP compared to the LP. A gene ontology analysis revealed that the most enriched GO terms were protein binding, multi-organism process, immune system and others. KEGG pathway analysis indicated that the differentially expressed genes were mostly enriched in Hippo signaling, insulin signaling, steroid biosynthesis and others. The analysis of the up- and downregulated genes provides important insights into the molecular events involved in lactogenesis, lactation and mammary gland development and will guide further research to enhance milk yield and optimize the constituents of yak milk., Competing Interests: The authors have declared that no competing interests exist.

Lactation is a physiological event that is exclusive to mammals. Lactation evolved as a strategy to improve the survival of the young by providing them with the complete nutrition that is required for survival upon birth as well as maternal-offspring bonding. Typically, milk production by the dam matches the demand of the young. The dairy cow is a unique exception in which the discoveries and genetic selection related to lactation physiology have been applied and resulted in a dramatic increase in milk yield of dairy cows. Studies on the role of mammary-derived serotonin and the coordination of various aspects of milk production and maternal metabolism have revealed novel mechanisms by which milk production and maternal metabolism can be improved. Furthermore, the investigation into molecular and cellular mechanisms regulating mammary gland function has revealed the importance of epigenetics on mammary gland function. Understanding mammary gland function at the cellular and physiological levels will be important for improving mammary gland control of maternal metabolism during early lactation. The early lactation period is a critical time for a dairy cow as that is when she is most susceptible to disease and metabolic disorders that can lead to negative effects on her productive capacity and overall health. Our research in the area of serotonin physiology has illustrated the importance of serotonin on the regulation of lactation and maternal homeostasis. Future research in the area of lactation physiology should be targeted at improving maternal health and longevity in the herd through manipulation of the signals the mammary gland sends to coordinate maternal metabolism and synthesize milk. Specifically, we believe that serotonin will play a central role in understanding the communication between the mammary gland and the maternal physiology during lactation., (Copyright © 2018 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

The objective of this study was to evaluate the influence of different amounts of rumen-undegradable protein (RUP) on intake, N balance, performance, mammary gland development, carcass traits, and hormonal status of Holstein heifers at different physiological stages (PS). Sixteen prepubertal (PRE) heifers (initial BW = 106 ± 7.6 kg; age = 4.3 ± 0.46 mo) and 16 pubertal (PUB) heifers (initial BW = 224 ± 7.9 kg; age = 12.6 ± 0.45 mo) were used in an experiment over a period of 84 d. Four diets with increasing RUP contents (38, 44, 51, and 57% of dietary crude protein) and heifers at 2 PS (PRE or PUB) were used in a 4 × 2 factorial arrangement of treatments in a completely randomized design. Throughout the experiment, 2 digestibility trials were performed over 5 consecutive days (starting at d 36 and 78) involving feed and ort sampling and spot collections of feces and urine. At d 0 and 83, body ultrasound images were obtained for real-time carcass trait evaluation. The mammary gland was ultrasonically scanned at d 0 and every 3 wk during the experiment. Blood samples were taken at d 0 and 84 to determine serum concentrations of progesterone, estrogen, insulin-like growth factor I (IGF-I), and insulin. No interaction between PS and the level of RUP was found for any trait. Apparent digestibility of dry matter, organic matter, and neutral detergent fiber corrected for ash and protein was not affected by RUP level but was lower for PRE compared with PUB heifers. Sorting against neutral detergent fiber corrected for ash and protein (tendency only) and for crude protein was greater for PUB than PRE heifers. Pubertal heifers had greater average daily gain (905 vs. 505 g/d) and N retention (25.9 vs. 12.5 g/d) than PRE heifers. In addition, average daily gain and N retention were greatest at 51% RUP of dietary protein. Mammary ultrasonography indicated no effects of RUP amounts on mammary gland composition, whereas PRE heifers had greater pixel values than PUB, indicating higher contents of fat rather than protein in the mammary glands of PRE heifers. Serum progesterone and IGF-I concentration was affected only by PS, and PRE heifers had greater values of progesterone and IGF-I concentrations than PUB heifers. Serum insulin concentration was unaffected by PS but tended to be higher at 51% of RUP. In conclusion, an RUP level of 51% increases body weight, average daily gain, feed efficiency, and N retention in heifers regardless of the PS. In addition, PRE heifers have a lower sorting ability and reduced intake, total-tract digestibility, and N retention. They also have higher amounts of fat in their mammary glands, even at moderate growth rates., (Copyright © 2018 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

Heat stress reduces cow milk yield and results in a significant economic loss for the dairy industry. During lactation, heat stress lowers milk production by 25 to 40% with half of the decrease in milk synthesis resulting from the reduced feed intake. In vitro studies indicate that primary bovine mammary epithelial cells display greater rates of programmed cell death when exposed to high ambient temperatures, which may lead to a decrease in the total number of mammary epithelial cells in the mammary gland, partially explaining the lower milk production of lactating cows under heat stress. The function of mammary cells is also altered by heat stress. In response to heat stress, mammary cells display higher gene expression of heat shock proteins, indicating a need for cytoprotection from protein aggregation and degradation. Further, heat stress results in increased gene expression without altering protein expression of mammary epithelial cell junction proteins, and does not substantially influence the integrity of mammary epithelium. These data suggest that the mammary gland strives to maintain cell-to-cell junction integrity by synthesizing more proteins to compensate for protein losses induced by heat stress. During the dry period, heat stress negatively affects mammary gland development by reducing mammary cell proliferation before parturition, resulting in a dramatic decrease in milk production in the subsequent lactation. In addition to mammary growth, the mammary gland of the heat-stressed dry cow has reduced protein expression of autophagic proteins in the early dry period, suggesting heat stress influences mammary involution. Emerging evidence also indicates that heifers born to cows that experience late-gestation heat stress have lower milk yield during their first lactation, implying that the maternal environment may alter mammary gland development of the offspring. It is not clear if this is due to a direct epigenetic modification of prenatal mammary gland development by maternal heat stress. More research is needed to elucidate the effect of heat stress on mammary gland development and function., (Copyright © 2018 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

Drying cows off at the end of lactation is a routine management practice in dairy operations. Most dairies in the United States and many other countries dry cows off abruptly (e.g., stop milking cows on a set day), which has been shown to affect cow comfort. Gradually reducing milk production is another approach to dry cows off, routinely used in some countries and herds. The objective of the study was to evaluate the effects of abrupt and gradual milk cessation and milk yield at the time on cow activity after dry-off. Daily lying time, number of lying bouts per day, average lying bout length, and steps taken per day by abruptly and gradually dried-off cows were monitored by data loggers for 2 wk before and after the final milking at the end of lactation. Gradual cows were milked once daily for the last week of lactation, and abrupt cows were milked as usual (3×/d) until the end of lactation. Gradual cessation of milking significantly reduced milk yield by the day of dry-off. After dry-off, gradual cows tended to have longer lying bouts than abrupt cows, but no other differences in cow activity between the 2 treatments were observed. Regardless of the dry-off method, the average length of a lying bout decreased by 4 min and total daily lying time decreased by 19 min after dry-off for each 5-kg increase in milk yield before dry-off. Lying behavior of primiparous cows was more affected by the level of milk yield at dry-off than that of older cows. A reduction in lying times with increasing milk yield may indicate discomfort due to the accumulating milk in the udder. Using a method that lowers milk production before dry-off and managing primiparous and multiparous cows separately around dry-off are beneficial for cow comfort after dry-off., (Copyright © 2018 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

Dairy milk consists of more than 85% water. Therefore, understanding the regulation of fluid absorption in the mammary gland is relevant to improving milk production. In recent decades, studies using different approaches, including blood flow, transmembrane fluid flow, tight junction, fluid flow of the paracellular pathway and functional mammary epithelial cell state, have been conducted aiming to investigate how mammary gland fluid absorption is regulated. However, the relationship between regulation mechanisms of fluid flow and milk production has not been studied systematically. The present review summarizes a series of key milk yield regulatory factors mediated by whole-mammary fluid flow, including milk, mammary blood flow, blood/tissue fluid-cell fluid flow and cell-alveolus fluid flow. Whole-mammary fluid flow regulates milk production by altering transporter activity, ion channels, local microcirculation-related factors, driving force of fluid transport (osmotic pressure or electrochemical gradient), cellular connection state and a cell volume sensitive mechanism. In addition, whole-mammary fluid flow plays important roles in milk synthesis and secretion. Knowledge gained from fluid flow-mediated regulatory mechanisms of the dairy mammary gland will lead to a fundamental understanding of lactation biology and will be beneficial for the improvement of dairy productivity. © 2017 Society of Chemical Industry., (© 2017 Society of Chemical Industry.)

Dietary Zn and heat stress alter gut integrity in monogastric animals. However, effects of Zn on mammary epithelial integrity in heat-stressed lactating dairy cows have not been studied. Multiparous lactating Holstein cows (n = 72) were randomly assigned to 1 of 4 treatments with a 2 × 2 factorial arrangement to study the effects of environment and Zn source on performance and mammary epithelial integrity. Treatments included 2 environments [cooled (CL) or not cooled (NC)] and 2 Zn sources [75 mg/kg of supplemental Zn as Zn hydroxychloride (IOZ) or 35 mg/kg of Zn hydroxychloride + 40 mg/kg of Zn-Met complex (ZMC)]. The experiment was divided into baseline and environmental challenge phases of 84 d each. All cows were cooled during the baseline phase (temperature-humidity index = 72.5), whereas NC cows were not cooled during environmental challenge (temperature-humidity index = 77.7). Mammary biopsies were collected on d 7 and 56 relative to the onset of environmental challenge to analyze gene expression of claudin 1, 4, and 8, zonula occludens 1, 2, and 3, occludin, and E-cadherin and protein expression of occludin and E-cadherin. Deprivation of cooling increased respiration rate (64.8 vs. 73.9 breaths/min) and vaginal temperature (39.03 vs. 39.94°C) and decreased dry matter intake (26.7 vs. 21.6 kg/d). Energy-corrected milk yield decreased for NC cows relative to CL cows (24.5 vs. 34.1 kg/d). An interaction between environment and Zn source occurred for milk fat content as CL cows fed ZMC had lower milk fat percentage than other groups. Relative to CL cows, NC cows had lower concentrations of lactose (4.69 vs. 4.56%) and solids-not-fat (8.46 vs. 8.32%) but a higher concentration of milk urea nitrogen (9.07 vs. 11.02 mg/mL). Compared with IOZ, cows fed ZMC had lower plasma lactose concentration during baseline and tended to have lower plasma lactose concentration during environmental challenge. Plasma lactose concentration tended to increase at 3, 5, and 41 d after the onset of environmental challenge in NC cows relative to CL cows. Treatment had no effect on milk BSA concentration. Cows fed ZMC tended to have higher gene expression of E-cadherin relative to IOZ. Compared with CL, NC cows had increased gene expression of occludin and E-cadherin and tended to have increased claudin 1 and zonula occludens 1 and 2 gene expression in the mammary gland. Protein expression of occludin and E-cadherin was unchanged. In conclusion, removing active cooling impairs lactation performance and affects gene expression of proteins involved in the mammary epithelial barrier, and feeding a portion of dietary zinc as ZMC improves the integrity of the mammary epithelium., (Copyright © 2018 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

The inhibition of prolactin release using cabergoline, a dopamine agonist, is an effective strategy to accelerate the changes in mammary secretion composition after drying-off. The objective of this study was to determine how cabergoline may affect mammary tissue remodeling during early involution. Holstein dairy cows were treated with either a single i.m. administration of 5.6 mg of cabergoline (Velactis, Ceva Santé Animale, Libourne, France, n = 7) or placebo (n = 7) at the time of drying-off. Mammary biopsy samples were collected 1 wk before drying-off (d -6), after 30 h of milk accumulation (d 1), and again 8 d following drying-off (d 8) to determine changes in gene expression, lactoferrin content, and cell turnover. Blood and mammary secretion samples were collected at d -6 and again at d 1, 2, 3, 4, 8, and 14 following the abrupt cessation of lactation to evaluate indicators of blood-milk barrier integrity and other markers of mammary tissue remodeling. Cabergoline induced less SLC2A1, BAX, CAPN2, and IGFBP5 mRNA expression. In contrast, cabergoline did not modify changes in cell proliferation and apoptosis. Following the cessation of lactation, changes in mammary secretion composition (Na + and K + ) and blood lactose concentrations were indicative of a loss in the blood-milk barrier function in both treatment groups. Cabergoline treatment affected only Na + and K + concentrations at d 1, suggesting a moderate increase in tight junction permeability. The increase in the activity of MMP9 and in mammary epithelial cell concentration in mammary secretions was greater in cabergoline-treated cows than in control cows, suggesting more mammary tissue remodeling. The increase in lactoferrin immunostaining in the mammary tissue occurred earlier for cabergoline-treated cows than for control cows, and was essentially localized in the stroma. Changes in some key markers of mammary involution suggest that cabergoline accelerates mammary gland remodeling. Thus, a single injection of cabergoline after the last milking would facilitate drying-off by enhancing mammary gland involution., (The Authors. Published by the Federation of Animal Science Societies and Elsevier Inc. on behalf of the American Dairy Science Association®. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/3.0/).)

The latter half of the 20th century and the early portion of the 21st century will be recognized as the "Golden Age" of lactation biology. This period corresponded with the rise of systemic, metabolomic, molecular, and genomic biology. It includes the discovery of the structure of DNA and ends with the sequencing of the complete genomes of humans and all major domestic animal species including the dairy cow. This included the ability to identify polymorphisms in the nucleic acid sequence, which can be tied to specific differences in cellular, tissue, and animal performance. Before this period, classical work using endocrine ablation and replacement studies identified the mammary gland as an endocrine-dependent organ. In the early 1960s, the development of RIA and radioreceptor assays permitted the study of the relationship between endocrine patterns and mammary function. The ability to measure nucleic acid content of tissues opened the door to study of the factors regulating mammary growth. The development of high-speed centrifugation in the 1960s allowed separation of specific cell organelles and their membranes. The development of transmission and scanning electron microscopy permitted the study of the relationship between structure and function in the mammary secretory cell. The availability of radiolabeled metabolites provided the opportunity to investigate the metabolic pathways and their regulation. The development of concepts regarding the coordination of metabolism to support lactation integrated our understanding of nutrient partitioning and homeostasis. The ability to produce recombinant molecules and organisms permitted enhancement of lactation in farm animal species and the production of milk containing proteins of value to human medicine. These discoveries and others contributed to vastly increased dairy farm productivity in the United States and worldwide. This review will include the discussion of the centers of excellence and scientists who labored in these fields to produce the harvest of knowledge we enjoy today.