The aim of the research reported in this Research Communication was to apply the 5-point body condition scoring (BCS) system to dairy buffaloes and subjectively validate it by assessing the intra- and inter-assessor agreement. For this purpose, the BCS system developed for dairy cows was applied to buffaloes. A total of 230 Nili Ravi buffaloes of varying parity, lactation stages and pregnancy status were enrolled from the Buffalo Research Institute, Pattoki, Pakistan. Four observers independently assigned BCS values to each enrolled buffalo in two phases, as follows: (1) during phase I, the assessors were trained for BCS assessment using a BCS chart developed by Elanco Animal Health Ltd.; and (2) during phase II, the assessors were trained using live buffaloes for BCS assessment. Kappa statistics (kw) were used to determine the intra- and inter-assessor agreement. The results revealed that the exact overall inter- and intra-assessor agreement was moderate (kw = 0.48–0.55) and increased to substantial levels after training on live animals (kw = 0.63–0.87). Furthermore, the intra- and inter-assessor exact agreement was higher (kw = 0.57–0.58) for buffaloes tied to the mangers compared to the buffaloes standing in the loafing area (kw = 0.50). The inter-assessor agreements within 0.25 and 0.5 points were almost perfect (kw = 0.97–1.0). The current results suggested that the 5-point BCS system (using a scale from 1 to 5 with 0.25 increments) had substantial agreement for assessment and repeatability when applied to buffaloes.

The experiment reported in this research paper aimed to evaluate the effects of high-starch or starch and oil-supplemented diets on rumen and faecal bacteria, and explore links between the structure of bacterial communities and milk fatty acid (FA) profiles. We used four Holstein dairy cows in a 4 × 4 Latin square design. Cows were fed a diet rich in cereals (high-starch diet with 23% starch content on dry matter (DM) basis), a diet supplemented with saturated FA from Ca salts of palm oil + 18% DM starch, a diet with high content of monounsaturated FA (from extruded rapeseeds) + 18% DM starch or a diet rich in polyunsaturated FA (from extruded sunflower seeds) + 17% DM starch. At the end of each experimental period, cows were sampled for rumen and faecal contents, which were used for DNA extraction and amplicon sequencing. Partial least squares (PLS) regression analysis highlighted diet-related changes in both rumen and faecal bacterial structures. Sparse PLS discriminant analysis was further employed to identify biologically relevant operational taxonomical units (OTUs) driving these differences. Our results show that Butyrivibrio discriminated the high-starch diet and linked positively with higher concentrations of milk odd- and branched-chain FA. YS2-related OTUs were key taxa distinguishing diets supplemented with Ca salts of palm oil or sunflower seeds and correlated positively with linoleic acid in milk. Similarly, diets modulated faecal bacterial composition. However, correlations between changes in faecal and rumen bacteria were poor. With this work, we demonstrated that high-starch or lipid-supplemented diets affect rumen and faecal bacterial community structure, and these changes could have a knock-on effect on milk FA profiles.

This research communication reports a study to model the growth curves for withers height (WH) and body weight (BW) to withers height ratio (BW:WH) using monthly records (from 1 to 24 months of age) for three breeds of dairy heifer (Holstein, Jersey and Brown Swiss). The data sets used were those reported by the Dairy Heifer Evaluation Project of Penn State Extension (USA) between 1991 and 1992. Four growth functions (monomolecular and Michaelis-Menten, both with diminishing returns behaviour, and Schumacher and López, both with asymptotic sigmoidal behaviour) were fitted using the non-linear regression procedure of the SigmaPlot software and the parameters estimated. The models were judged for goodness of fit using adjusted coefficient of determination ($R_{{\rm adj}}^2 $), root mean square error (RMSE), Akaike's information criterion (AIC) and Bayesian information criterion (BIC). Assessing the goodness of fit by $R_{{\rm adj}}^2 $ (>0.99 in all cases) reveals the generally appropriate fit of the models to the data. The non-sigmoidal functions (i.e. Michaelis–Menten and monomolecular) provided the best fits giving the lowest values of RMSE, AIC and BIC. Based on the chosen statistical criteria, the Schumacher and López equations provided acceptable fits to the WH and BW:WH growth curves, but showed points of inflexion at times before birth, indicating that these growth curves are not sigmoidal. In conclusion, evaluation of the different non-linear growth functions used in this study indicated their potential for modelling growth patterns in dairy heifers.

The objective of the study was to describe the feeding behaviour of primiparous and multiparous Jersey cows compared to Holstein cows housed in separate groups in the same barn. Such information could help farmers to optimise management with respect to welfare and production. Yet, it remains limited for Jersey cows over the entire period of lactation. Feeding data of 116 Danish Jersey (mean parity 2.14 ± 1.32) and 218 Danish Holstein cows (mean parity 1.90 ± 1.16) were assessed using automatic feeders from day 15 to 252 of lactation. Total eating duration, duration of eating per visit, intervals between meals, number of visits per day and the eating rate were analysed using linear mixed effects models. The cows were kept in a loose-housing system, with cubicles and automatic milking robots, and the group composition was dynamic. Compared to Holstein cows, Jersey cows visited the feeder significantly more often with shorter between meal intervals. However, the visit duration and total daily eating time and eating rates were significantly shorter for Jersey cows. There was no difference between breeds in the daily eating time and eating rate of older cows. Younger Jersey cows had significantly lower eating rates than older Jersey cows. No other difference in parity was found within Jersey cows. Weeks in milk significantly affected the eating time per day, number of visits per day and eating rate. The trajectories of outcome variables during lactation did not differ between the two breeds. In conclusion, we found substantial differences in the feeding behaviour of Jersey and Holstein cows, however, these differences could also be related to a group effect.

The aim of this research communication was to examine and report the current situation of dairy farms in the province of Lugo (Galicia, Spain) regarding facility problems. We assessed the facilities of 168 free-stall dairy farms, housing in total 9228 Holstein cows in milk. Housing factors related to the resting area, circulation area, feeding area, ventilation area and milking area, as well as animal-handling features, were evaluated. Distance measurements were performed using a laser metre or a roll metric tape. A survey was conducted to gather information about cleaning and preventive protocols. Our results showed that most farms do not comply with the objective for cubicle measurements, width of the crossovers, type of flooring and presence of a quarantine pen, which may have a negative impact on the health and productivity of the animals. Therefore, to maximise the profits of the farm, the recommendations given in this study may be useful as a guide when building a new farm or remodelling the existing ones.

This research paper aimed to evaluate the association between feeding waste milk to calves and the occurrence of antimicrobial multi-resistance by extended spectrum β-lactamase (ESBL) enzymes through determining their production by E. coli isolates from 32 dairy farms. Among β-lactamase enzymes, ESBL provide resistance to a wide variety of β-lactam antimicrobials including penicillin and 2nd, 3rd and 4th generation cephalosporins. Feeding waste milk to calves has been observed to lead to increased antimicrobial resistance in faecal isolates of calves. In each farm included in this study, faecal samples were collected from the rectum of five healthy calves in the first month of life and pooled into a single container. Five isolates from each pool were selected and confirmed to be E. coli by amplification of the 16S rRNA gene. ESBL production was confirmed phenotypically on 148 isolates from 31 farms by use of the double-disk synergy test. Genotypic confirmation of ESBL production was performed by PCR for the genes blaCTX-M-1, −2, −8, −9 and blaCMY-2. A questionnaire was also performed and a mixed logistic regression model was used to identify risk factors for the occurrence of antimicrobial resistance. A negative binomial regression model was also used, in order to assess whether there was any association between certain farm management practices and the number of ESBL-producing E. coli isolates from each farm. Phenotypic confirmation of ESBL production was obtained on 40 E. coli isolates from 15 farms (48.4%), whereas genotypic confirmation was obtained on 55 isolates from 20 farms (64.5%). The use of three or more different intramammary antimicrobials to treat mastitis within the previous year significantly impacted the number of ESBL-producing E. coli isolates; on farms that did so, there were more isolates in which ESBL-producing E. coli was present, when compared to farms that had used less formulations within the same time span.

We investigated whether prepartum levels of serum bone biomarkers are related to the degree of parturient hypocalcaemia and risk of milk fever (MF) in dairy cows with advancing parity. A total of 58 late-pregnant cattle were assigned to four groups: nulliparous, primiparous, multiparous in the 2nd lactation and multiparous in the 3rd–5th lactation. The multiparous cows were further assigned to MF and non-MF groups according to the onset of MF. Serum samples were obtained from the cows during the 3 weeks prepartum to 5 d postpartum period for the measurement of serum calcium (Ca) and three bone biomarkers: tartrate-resistant acid phosphatase isoform 5b (TRAP5b), osteoprotegerin (OPG) and bone isoenzyme of alkaline phosphatase (ALP3). The ratios of OPG to TRAP5b (O/T ratio) and ALP3 to TRAP5b (A/T ratio) were calculated. The data from all cattle showed that the severity of hypocalcaemia at parturition increased with advancing parity/age. The MF cows had elevated serum TRAP5b activity and a decreased O/T ratio after parturition, suggesting an increased number of osteoclasts due to osteoclastogenesis, in response to severe hypocalcaemia. The MF cows showed lower serum ALP3 activity during the 3 weeks prepartum than the non-MF cows, therefore, prepartum osteoblast function was likely weak in the MF cows. During the 2–3 weeks prepartum, serum ALP3 activity and the A/T ratio had moderate associations with the serum Ca concentration at day 0 (day of calving) in the multiparous cows, and receiver operating characteristic curve analysis revealed that ALP3 activity had excellent ability to predict MF. In conclusion, prepartum serum ALP3 activity is a promising biomarker to predict MF in multiparous cows.

We investigated the effect of different types of mother-young contacts (fenceline, restricted and no contact) on the health, growth performance, behaviour and physiological state of Murrah buffalo calves. A total of 24 calves were allocated to three groups of 8: NCM (no calf–mother contact), RCM (restricted calf–mother contact) and FCM (fenceline calf–mother contact). At three months of age, the FCM calves had a higher average body weight (58.6 ± 1.5 kg) than the RCM (52.8 ± 1.3 kg) and NCM (53.6 ± 1.3 kg) calves (P < 0.05). The average daily gain (ADG) at three months of age was greater in FCM (0.6 ± 0.1 kg/d) than RCM (0.5 ± 0.1 kg/d) and NCM (0.5 ± 0.1 kg/d) calves (P < 0.05). The mean immunoglobulin G (IgG) concentrations were significantly greater on d 7, 28, 42 and d 56 of sampling (P < 0.05 for all comparisons) in FCM than RCM and NCM calves. The mean cortisol levels were highest in the NCM calves followed by RCM and the lowest values were observed in FCM calves on d 0, d 7, d 28, d 42 and d 56, respectively (P < 0.05, for all recorded days). The diarrhoea score and eggs per gram of faeces (EPG) were significantly greater in the NCM group compared to the RCM and FCM groups. The time spent in cross sucking, licking inanimate objects and self-licking was greatest (P < 0.05) in NCM calves, followed by RCM calves, and was almost non-existent in the FCM group on all the recorded weeks. Time spent in backward ear position was greater (P < 0.05) in FCM than RCM and NCM calves. The average time spent in forward ear position was greatest (P < 0.05) in the RCM calves followed by NCM with the lowest time observed in FCM calves. The mean duration of time spent by calves in the asymmetrical and axial ear position differed significantly (P < 0.05) among NCM, RCM and FCM calves, P < 0.05. The full and fenceline buffalo calf–mother contact system showed higher growth rates, lower levels of stress responses, oral stereotypies and eggs per gram of faeces. In addition, ear postures may be used as a dependable, and reliable measure of positive, low arousal emotional states.

The practices adopted in dairy farms can positively or negatively affect the perception of consumers. To meet consumer expectations and improve the productivity of dairy farms, a welfare certification system has recently been initiated in Brazil. In this research communication we describe the perceptions of Brazilian consumers and farmers regarding the implementation of welfare certification systems and the most common practices that affect animal welfare on dairy farms. For this purpose, two semi-structured questionnaires were used: one applied to 409 consumers and the other to 158 dairy farmers. The results demonstrate that consumers are concerned with the adoption of welfare practices in animal husbandry at dairy farms, mainly on topics related to movement restriction and cow-calf separation. Thus, the majority of consumers state that they are willing to pay more for welfare-certified dairy products. In addition, most dairy farmers are interested in adopting a welfare certification system, especially if it could add value to the raw milk sold to industries. Veterinarians and animal scientists are important for disseminating animal welfare recommendations, and the consequences of its improper adoption need to be emphasized. Finally, dairy farms need improvements regarding environmental hygiene, thermal conditions, animal husbandry, health, and milking processes. In conclusion, consumers and farmers are interested in welfare systems and their certification, and there is a need for stakeholders to make welfare certification a reality in the Brazilian dairy supply chain.

The results reported in this research communication aimed to reduce the housing discomfort and mastitis incidence of lactating Jersey crossbred cows through modifications to the roof and floor of loose housing stalls. The experiment was conducted on twenty Jersey crossbred cows and they were distributed equally into two different types of housing: (i) existing shed/control group (T0) – concrete floor and asbestos roof and (ii) modified shed/treatment group (T1)- sand flooring (4–6 inch deep; 38% of total area) and a thatch ceiling (4″ thick) under an asbestos roof. Under-roof surface temperature differed significantly (P < 0.01) between the two sheds during both the winter season but a greater difference was observed in the summer (modified sheds were 5–9 °C cooler than the control shed during peak hours (10 am to 3 pm) of the day. The milk yield (kg/d) in both seasons was significantly higher in the treatment group than in the control group (P < 0.01). The overall milk fat and total solid percentage were significantly higher in the treatment than the control group (P < 0.01) but solids not fat (SNF) did not show any difference between the groups. The percentage of milk fat did not differ significantly between the seasons, while the SNF and total solids were also significantly higher in the winter than the summer season (P < 0.01). There was a significantly lower subclinical mastitis test score (MCMT grade: P < 0.05) in the treatment group than in the control group. In the treatment group, the somatic cell count (SCC) was numerically but non-significantly less than in the control group. No differences in MCMT and SCC were observed between seasons (P > 0.05). It was concluded that the provision of sand as stall flooring and an under-roof thatch ceiling as a heat insulator was significantly associated with increased milk yield, milk composition and possibly, lower somatic cell count in dairy Jersey crossbred cows. However, since the study was limited to a single replicate of each housing system with analysis done at individual cow level, further work is needed to confirm these conclusions.

To maintain and enhance cow productivity and welfare, it is important that we can accurately assess and understand how cows respond to the physiological demands of gestation and lactation. Several methods have been developed for assessing the physiological responses to stressors and for detecting distress in cattle. Heart rate (HR) variability (HRV) is a non-invasive measure of autonomic nervous system activity and consequently a component of the physiological response to stress. In cattle, HRV has been successfully used to measure autonomic responses to a variety of health conditions and management procedures. The objectives of this study were to determine whether, among commercial Holstein Friesian cows and across farms, relationships exist between cow-level factors, HR and HRV. HRV parameters were compared with production records for 170 randomly selected, Holstein-Friesian-cows on 3 commercial dairy farms. Production data included parity, days in milk (DIM), milk yield, somatic cell count (SCC), % butterfat and protein, body condition score (BCS) and genetic indices. Fixed-effect, multivariable linear regression models were constructed to examine the association between cow-level variables and HRV parameters. Statistically significant relationships were found between HR and farm, temperature and BCS, and between HRV parameters and farm, rectal temperature, BCS, DIM, and percentage butterfat. Given the significant association between farms and several of the indices measured, it is recommended that care must be taken in the interpretation of HRV studies that are conducted on animals from a single farm. The current study indicated that within clinically normal dairy cattle HRV differed with the percentage of butterfat and BCS. Based on the relationships reported previously between HRV and stress in dairy cattle these results suggest that stress may be increased early in lactation, in cows with BCS <2.75 that are producing a high percentage of butterfat milk. Future work could focus on the physiological mechanisms through which these factors and their interactions alter HRV and how such physiological stress may be managed within a commercial farm setting.

This research communication reports the relative abundance of fatty acids in mammary fat pad (MFP) and subcutaneous adipose (SCA) tissues for Holstein heifer calves receiving 0, 3, or 12 daily injections of estradiol immediately prior to tissue collection. The objective of this study was to determine if the MFP and SCA fatty acid profiles were affected by estradiol administration and if such a response differs between adipose tissue depots. Twelve Holstein heifer calves were reared on a common diet and administered 12 daily injections prior to euthanasia. Injections were either daily injections of corn oil (n = 4; CON), 9 injections of corn oil followed by 3 injections of estradiol (n = 4; SHORT), or 12 injections of estradiol (n = 4; LONG). Fatty acids were extracted from collected MFP and SCA tissues samples and analyzed using gas chromatography. The MFP tissues contained a greater abundance of saturated fatty acids than SCA tissues which complemented a reduced abundance of mono-unsaturated fatty acids in the MFP than SCA. Extended duration of estradiol administration increased the abundance of total omega 3 fatty acids in both MFP and SCA tissues. There was a treatment by tissue interaction for several of the C18:1 and C18:2 isomers indicating that estradiol's effects on fatty acid uptake and metabolism are tissue specific. Additionally, C18 uptake and metabolisms may have important roles in mammary growth and development. Together, these results indicate that the MFP responds differently to estradiol administration than SCA tissues and that these alterations are associated with different degrees of induced mammary growth via estradiol.

The objective of this retrospective longitudinal study was to evaluate the relationship between dry period length and the production of milk, fat, protein, lactose and total milk solids in the subsequent lactation of Holstein dairy cows under tropical climate. After handling and cleaning of the data provided by the Holstein Cattle Breeders Association of Minas Gerais, data from 32 867 complete lactations of 19 535 Holstein animals that calved between 1993 and 2017 in 122 dairy herds located in Minas Gerais state (Brazil) were analysed. In addition to dry period length, calving age, lactation length, milking frequency, parity, calf status at birth, herd, year, and season of calving were included in the analysis as covariables to account for additional sources of variation. The machine learning algorithms gradient boosting machine, extreme gradient boosting machine, random forest and artificial neural network were used to train models using cross validation. The best model was selected based on four error metrics and used to evaluate the variable importance, the interaction strength between dry period length and the other variables, and to generate partial dependency plots. Random forest was the best model for all production outcomes evaluated. Dry period length was the third most important variable in predicting milk production and its components. No strong interactions were observed between the dry period and the other evaluated variables. The highest milk and lactose productions were observed with a 50-d long dry period, while fat, protein, and total milk solids were the highest with dry period lengths of 38, 38, and 44 d, respectively. Overall, dry period length is associated with the production of milk and its components in the subsequent lactation of Holstein cows under tropical climatic conditions, but the optimum length depends on the production outcome.

This research communication evaluates experts’ opinions on the importance and weights of six gait aspects. In 2016, a Qualtrics (Qualtrics LLC., Provo, Utah) survey was distributed to lameness experts. Six gait aspects – general symmetry, tracking, spine curvature, head bobbing, speed and abduction as well as adduction were included. Respondents were asked to rank the gait aspects from 1 (most important) to 6 (least important), and to indicate which weight each gait aspect should receive when assessing lameness. For each gait aspect, frequency (percentage %) was used to describe the distribution of rank, and medians as well as 25th and 75th percentiles were used to summarize assigned weights. Thirty-nine percent of respondents ranked general symmetry first, followed by 32% for tracking, and 19% ranked spine curvature third. Head bobbing ranked fourth with 10% whereas, speed, abduction and adduction were not ranked. The median, 25th and 75th percentiles weight for each gait aspect were: general symmetry (25, 15, and 30), tracking (20, 10, and 30), spine curvature (20, 10, and 21), head bobbing (15, 10, and 20), speed (10, 5, and 20), and abduction and adduction (10, 5, and 10). General symmetry and tracking were deemed the most important gait aspects. A composite gait score can be calculated based on weighted importance of different gait aspects to indicate possible lameness.

This study is the first report to investigate the relationships between time of parturition and milk productivity and quality, as well as indices related to udder measurements and meteorological variables, in Saanen goats raised under semi-intensive conditions. Goats giving birth in the hours of darkness had higher milk production than those that gave birth in the hours of daylight, while those giving birth during the evening hours had lower somatic cell count (SCC) than those with parturition during the daylight and night hours (P < 0.05). In addition, the time of parturition was associated with rear udder depth, udder circumference, and udder volume traits (P < 0.01). Parity and time of parturition × parity interaction had significant effects on lactation milk yield and lactation length, as well as milk fat, protein, lactose, total solids content and electrical conductivity (P < 0.05 to P < 0.01). The lactation stage, daily milk yield level and parity affected milk SCC (P < 0.05). Ambient temperature and daylight length had strong effects on daily milk yield (P < 0.05). These findings have practical implications for productivity, quality and health promotion efforts aimed at increasing Saanen goat dairy productivity consistently in the face of climatic changes in a semi-intensive system.

This study aimed to evaluate the effect of an extended voluntary waiting period (VWP) on SCC, SCC elevations and clinical mastitis incidence during the complete lactation and the first 6 weeks of the next lactation. Holstein-Friesian dairy cows (N = 154) were blocked for parity, expected milk yield, calving season and breeding value for persistency and were randomly distributed across 3 VWP (50, 125, or 200 d: VWP-50, VWP-125, VWP-200). Cows were monitored from calving until 6 weeks into the next lactation, or until culling. An elevation of SCC in milk was defined as SCC in milk ≥200 000 cells/ml after two previous weeks with SCC < 200 000 cells/ml. Over the complete lactation, extending the VWP did not affect SCC elevations and the occurrence of clinical mastitis per lactation or per cow per year. There was no clear effect of VWP length on SCC in the complete lactation, except that multiparous cows in VWP-125 had a higher SCC compared with multiparous cows in VWP-50. Dry-off antibiotic usage per cow per year was lower in VWP-200 compared with VWP-50 for multiparous cows. In the first 6 weeks of the next lactation, cows in VWP-200 had a higher SCC compared with cows in VWP-50, with no effect of VWP on the number of elevations of SCC or the occurrence of clinical mastitis. Extending the VWP may therefore be used to reduce the frequency of transition periods and the associated use of dry-cow antibiotics, with limited impact on udder health, and a similar occurrence of SCC elevations and clinical mastitis per year.

Plasminogen (Pg) activation on the cell surface is important for various (patho)physiologic conditions, and Plg-RKT is a cell membrane protein that binds to Pg and promotes its activation. To evaluate the role of Plg-RKT in atherosclerosis, Plgrkt gene in Ldlr−/−/Apobec1−/− was modified using in vivo CRISPR/Cas9. Synthetic RNA for Plgrkt and Cas9 complex was electroporated into the fertilized eggs in the oviducts. Plgrkt deficient mice were established through a 1-bp deletion, and in this research communication we report their lactational ability. In contrast to Plgrkt−/− mice developed by a conventional method, these newly developed mice did not suffer lactation failure and could maintain their pups until weaning. The major obvious difference between these lines is the area of gene modification. The conventionally developed mouse possesses about 10 kb deletion of Plgrkt, which might relate to the lactation failure. Lactation failure is a lethal phenotype in mammals, and analyses of causative genes are especially important for dairy industries. Further genome-wide analyses with both Plgrkt−/− mice may help to establish causative genes for lactation failure.

This research paper addresses the hypothesis that stress, induced by ACTH administration and cortisol release increases somatic cell count (SCC) in mammary secretion, and improves the effectiveness of dry off in goats. We report indicators of milk synthesis and mammary gland involution during dry off. Thirty Saanen goats were subjected to abrupt dry off and treatments: (1) ACTH administration (ACTH) or (2) placebo (Control) on days 1, 3, 6, 9, 12, 15, 30, and 60 of dry off. The expression of target genes in mammary tissue that are related to milk synthesis and cell survival such as insulin-like growth factor 1 receptor (IGF1R), phosphatidylinositol-3-kinase (PIK3CA), protein kinase B (AKT1) and mechanistic target of rapamycin (MTOR), casein (CSN2), lactalbumin (LALBA) and lactoferrin (LF) were evaluated, and plasma cortisol concentration, SCC, leucocyte count, and microbiological analyses in milk and mammary secretions were assessed. ACTH significantly downregulated the expression of IGF1R and upregulated the expression of PIK3CA in mammary tissue, increased lactoferrin concentration and SCC, and changed immune cell levels in mammary secretions compared to Control. Furthermore, ACTH administration increased the percentage of dry goats compared to the Control (73 vs. 46%, respectively). We conclude that the effect of stress via ACTH administration and cortisol release accelerated mammary involution during the early dry-off period.

We evaluated the relationship between plasma levels of anti-Müllerian hormone (AMH) and the number of antral follicles at the restart of the follicular wave in crossbred Holstein cows reared under extensive grazing systems over 2500 m above sea level. The study included 140 cows from 15 farms that were in average at the 75.3 ± 2.10 d post partum. Animals were synchronized according to the following regime: day 0 = intravaginal progesterone releasing device (IPD) + estradiol benzoate (EB); day 7: withdrawal of IPD + prostaglandin; and day 8: EB, for restart of the follicular wave on day 11. On this day 11, antral follicle counts (AFCs) were made by transrectal ultrasound, and a plasma sample was taken for the determination of AMH. The mean AMH plasma level was 0.06 ± 0.03 ng/ml and the mean AFC was 17.26 ± 0.38 follicles. A strong positive linear correlation was found between these two variables (r = 0.783, r = 0.613, P < 0.0001). Cows were categorized according to AMH concentration as high (>0.09 ng/ml), intermediate (0.09–0.05 ng/ml) or low (<0.05 ng/ml). Cows with high AMH presented a higher AFC (25.0 ± 2.21 follicles) than those with low AMH (14.08 ± 2.68 follicles; P < 0.001. Our results suggest that the cut-off value of AMH = 0.09 ng/ml may be useful for selecting donors in multiple ovulation embryo transfer programs involving cows with these characteristics. Our data further suggest that AMH plasma concentration correlates with AFC and can be used as an endocrine biomarker of the number of antral follicles present at a given moment of the estrous cycle in crossbred Holstein cows raised at altitudes above 2500 m.

The goal of the present study was to determine the best quarter milk flow for teat cup detachment depending on lactational stage and milking interval to optimize the milking process in automatic milking systems (AMS). Milking characteristics and post-milking teat condition were recorded in an AMS with all actions from teat cleaning to post-milking teat dipping occurring in the same teat cup and liner (GEA DairyRobot R9500). In 24 dairy cows, 12 in early (<80 DIM) and 12 in late lactation (>180 DIM), 294 milkings were recorded during 12 consecutive days. Teat cup detachment was set at a quarter milk flow of 50, 87.5 or 125 g/min. Voluntary milking intervals varied similarly in cows in early (6.1–14.8 h) and late lactation (6.3–15.7 h). Total milk yield, milk production per h and average milk flow were higher in early than in late lactation cows. Total milk yield per milking did not differ between detachment levels. The mean milking time was reduced by up to 1.5 min at the highest compared to the lowest detachment level mainly in early lactation cows (P < 0.05). However, no significant effect of the detachment level on milking time was observed at milking intervals >10 h. Average milk flow was higher at milking intervals >10 h than ≤10 h (P < 0.05). In the early lactation cows (only) the average milk flow increased with higher detachment levels (P < 0.05). Teat condition did not differ among detachment settings or milking intervals. In conclusion, teat cup detachment up to 125 g/min reduces milking time in both early and late lactation without a loss of milk yield or affecting the teat condition. Combined with a higher average milk flow through avoiding too short milking intervals, the total milking time and hence stall occupancy can be optimized by early teat cup detachment.