This study investigated the age at first calving in Holstein heifers and its impact on various production parameters. A sample of 737 Holstein heifers born between 2015 and 2018 and finishing their first lactations between 2018 and 2020 was included. Cluster analysis revealed three groups based on age at first calving: high precocity, medium precocity and low precocity. Medium-precocity group exhibited the highest 305-day milk yield and peak milk production. Additionally, the same group demonstrated superior mean production per lactation, 305-day milk protein content, and 305-day milk yield. The lowest somatic cell count was found in the low-precocity group. In conclusion, medium-precocity cows showed better results such as higher 305-day milk yield and peak milk production.

The present study assessed if salivary crystallization pattern (ferning pattern formed as a result of the higher levels of salt content in the dried sample) could be used for estrus detection and for diagnosis of pregnancy/non-pregnancy in dairy cows. Saliva and blood samples were collected from non-pregnant cycling cows (Sahiwal breed; n = 20) on alternate days from the day of estrus till next estrus. Then, all the cows were inseminated and saliva and blood sampling were continued further for a period of 22 d post-insemination. Pregnancy diagnosis was carried out on day 45 post-insemination and eight cows were found to be pregnant. The salivary crystallization pattern and estradiol:progesterone ratio during estrous cycle and during pregnancy were compared among these cows. Six types of salivary crystallization patterns were discerned; distinct patterns such as branch-like, fern-like, fir-like and combinations of these. Fern-like pattern was observed in all the cows on the day of estrus (first measurement day) and furthermore, all of the cows that subsequently became pregnant had fern-like salivary crystallization pattern at the time of insemination. Saliva of all the pregnant cows showed branch-fir type of crystallization pattern on day 16 post-breeding while only 50% of non-pregnant cows showed this pattern on day 16 of estrous cycle. The appearance of fern-like pattern was positively and significantly related to estradiol:progesterone ratio (r = 0.86; P < 0.001). The findings were validated on a separate group of cycling cows (n = 32). We can conclude that salivary crystallization pattern might serve as a non-invasive and cost effective and easy-to-use cow-side tool for estrus detection and early pregnancy/non-pregnancy diagnosis in cows upon validation on a larger sample size.

Live weight (LW) is an important piece of information within production systems, as it is related to several other economic characteristics. However, in the main buffalo-producing regions in the world, it is not common to periodically weigh the animals. We develop and evaluate linear, quadratic, and allometric mathematical models to predict LW using the body volume (BV) formula in lactating water buffalo (Bubalus bubalis) reared in southeastern Mexico. The LW (391.5 ± 138.9 kg) and BV (333.62 ± 58.51 dm3) were measured in 165 lactating Murrah buffalo aged between 3 and 10 years. The goodness-of-fit of the models was evaluated using the Akaike Information Criterion (AIC), Bayesian Information Criterion (BIC), coefficient of determination (R2), mean-squared error (MSE) and root MSE (RMSE). In addition, the developed models were evaluated through cross-validation (k-folds). The ability of the fitted models to predict the observed values was evaluated based on the RMSEP, R2, and mean absolute error (MAE). LW and BV were significantly positively and strongly correlated (r = 0.81; P < 0.001). The quadratic model had the lowest values of MSE (2788.12) and RMSE (52.80). On the other hand, the allometric model showed the lowest values of BIC (1319.24) and AIC (1313.07). The Quadratic and allometric models had lower values of MSEP and MAE. We recommend the quadratic and allometric models to predict the LW of lactating Murrah buffalo using BV as a predictor.

Sensors are increasingly being used to monitor animal behaviour. Data handling methods have, however, lagged behind the continuous data stream to some extent, often being limited to summarizing data into daily averages at group level. This research reflection presents our opinion of the neglected application of 24-h pattern analysis. Recent studies of dairy cow behaviour have demonstrated that additional ways of analysing data improve our understanding of animal behaviour and add value to data that were already retrieved. The terminology for the described 24-h patterns differs between these studies, making them difficult to compare. Thus, diurnal, circadian, daily, periodicity and 24-h pattern are all terms used to describe dairy cow activities over a 24-h period. Several studies have shown that the 24-h behavioural pattern at herd level is relatively consistent over time, and that with well-established management routines, a specific herd signature will be evident. However, within a herd, individual cows may have individual 24-h patterns with more or less variability. Recent studies suggest that deviations from herd and/or individual 24-h patterns can be used to describe cow robustness, as well as to predict disease. We strongly believe that individual and herd 24-h patterns provide a great deal of information about behaviour and that these patterns offer opportunity for more precise and timely health management and welfare monitoring.

In dairy production, high feed efficiency (FE) is important to reduce feed costs and negative impacts of milk production on the climate and environment, yet little is known about the relationship between FE, eating behaviour and activity. This research communication describes how cows differing in FE, expressed as daily energy corrected milk production per unit of feed intake, differed in eating behaviour and activity. We used data from a study of 253 lactations obtained from 97 Holstein and 91 Jersey cows milked in an automatic milking system. Automated feed troughs recorded feed intake behaviour and cows wore a sensor that recorded activity from 5 to 200 d in milk (DIM). We used a mixed linear model to estimate random solutions for individual cows for traits of steps, lying and eating behaviour and calculated their correlation with FE during four periods (5–35, 36–75, 76–120 and 121–200 DIM). Separate analyses were performed for each breed and period. We found that individual level correlations between FE and behaviour traits were stronger in Jersey than in Holstein cows. Eating rate correlated weakly negatively to FE in Holstein cows and more strongly so in Jersey cows, such that efficient Jerseys were slower eaters. The physical activity of Jersey cows was weakly and negatively correlated to FE, but this was not the case in Holstein cows. We conclude that eating rate was consistently negatively associated with FE throughout lactation for Jersey cows, but not for Holstein cows.

Exploration of novel strategies for early pregnancy diagnosis is pivotal in enhancing the reproductive potential and monetary gains from dairy herds. In buffalo, the trophectoderm cells of the elongating conceptus secrete interferon-tau that stimulates the transcription of various genes in peripheral blood mononuclear cells (PBMC) during the peri-implantation period. We explored the differential expression of classical (ISG15) and novel (LGALS3BP and CD9) early pregnancy markers in PBMC of buffaloes during various stages of pregnancy. Natural heat was detected in buffaloes by assessing the vaginal fluid, and artificial insemination (AI) was done. Whole blood was collected from the jugular vein in EDTA-containing vacutainers for PBMC isolation before AI (0-day) and 20, 25 and 40 d post-AI. On day 40, transrectal ultrasonography examination was performed to confirm pregnancy. The inseminated non-pregnant animals served as control. Total RNA was extracted using the TRIzol method. The temporal abundance of ISG15, LGALS3BP and CD9 genes in PBMC was compared between pregnant and non-pregnant groups (n = 9 per group) using real time-qPCR. Results showed transcripts of ISG15 and LGALS3BP were more abundant at 20 d in the pregnant group compared to the 0 d and 20 d values of the non-pregnant group. However, due to variability in expression, threshold (Ct) cycle of RT-qPCR alone could not distinguish pregnant and non-pregnant animals. In conclusion, ISG15 and LGALS3BP transcripts abundance in PBMCs are potential candidate biomarkers for early prediction of buffalo pregnancy 20-days post-AI, but further work is required to allow the development of a reliable new methodology.

The aim of this study was to evaluate the effect of the sprinkler cycle and flow rate on physiological, behavioural, and productive responses in dairy buffaloes. Nine Nili Ravi lactating buffaloes were subjected to three sprinkler cycles and two flow rates using a double replicated 3 × 3 Latin square design. The flow rates were 1.25 and 2 l/min, and the sprinkler cycles (minutes water on/off, number of cycles/h) were: 3/3, 10 cycles; 3/6, 7 cycles and 3/9, 5 cycles. The showering was applied from 0800 till 1630 h daily. In the first square of 21 d, each of the three sprinkler cycles was applied using a 1.25 l/min flow rate for 7 d per cycle. In the later square, the same treatments (sprinkler cycles) were applied using the 2 l/min flow rate. The average temperature humidity index during the study period was 85.7 ± 3.8 (Mean ± sd). The result showed that the 3/3 treatment group had lower body temperature and respiration rate than the other groups. The buffaloes in the 3/3 group produced 0.5 and 0.7 kg more milk with 1.4 and 2.4% more fat than the 3/6 and the 3/9 treatment groups, respectively. Similarly, the 2 l/min flow rate had a lower core body temperate and respiration rate and higher milk yield than the 1.25 l/min group. The 3/3 showering cycle with a 2 l/min flow rate appeared effective in improving physiological responses and milk yield in dairy buffaloes.

In this short commentary I recall a long-term experiment that was sketched out to determine if the low incidence of mammary cancer in dairy animals reflects a low incidence in these species generally or is the result of a protective effect of early pregnancy and long lactations. Although that experiment was never done, I discuss these questions in the light of developing knowledge on the incidence of cancer in ruminants generally and in the mammary gland in particular.

This Research Communication investigates how well U.S. dairy farmers understand the voting behaviour and willingness to pay of consumers for products with production traits relevant to animal health, welfare and biotechnology. Accurately understanding consumer behaviour is key to making sound production decisions and reducing risks. Comparing survey data with the literature shows that U.S. dairy farmers correctly assess consumer attitudes and behaviour over animal welfare practices like pain-controlled dehorning but could improve knowledge of attitudes towards antibiotic use and novel biotechnologies like gene editing.

We compared the virulence profile and REP-PCR genotypes of Escherichia coli strains isolated from subclinical and clinical mastitis cases and dairy farm environments in Minas Gerais State, Brazil, to determine virulence factors and genotypes potentially associated with subclinical persistence in the udder. The virulence profile was obtained by the search for three virulence genes: lpfA (long polar fimbriae), fliC (flagella), and escN (type III secretion system). Subclinical isolates exhibited mainly the fliC gene (33.33%) and fliC + escN genes (30.30%). Clinical isolates exhibited mainly fliC + escN genes (50%) and environmental isolates the lpfA + escN genes (58.04%). Strains isolated from subclinical mastitis showed 6.75 times more positivity to fliC than environmental isolates. Thirty-four genotypes were observed in the REP-PCR analysis, and clinical mastitis isolates indicated more genetic proximity to dairy farm environment isolates than subclinical mastitis isolates. In conclusion, the results suggested that flagella may be an important virulence factor for mammary persistent E. coli infection in cattle, however, none of the E. coli REP-PCR genotypes were associated with subclinical infection.

Mammary lipid secretion is generally held to be unique and remarkably uniform between the many different orders of mammals. It produces a unit membrane-bounded milk fat globule (MFG). The unit membrane is separated from the lipoprotein boundary of what was the cytoplasmic lipid droplet (CLD) boundary by a uniform layer of cytoplasmic proteins. In 3–8% of the MFG in all species examined this cytoplasmic layer widens to include cytoplasmic organelles which are referred to as ‘crescents’. This defines the MFG secretion as apocrine indicating a closely regulated process which minimises the loss of mammary epithelial cell (MEC) cytoplasm. The apocrine nature of the secretion might be expected since the evolution of the mammary gland is considered to be from an apocrine secreting skin gland. This short Research Reflection review is designed to investigate the exact cytoplasmic interactions which allow such efficient lipid secretion. There are two main scenarios: one which assumes that the observed close association between CLD and GV results in the CLD being released as a consequence of sequential exocytosis of the content of the associated GV. The second assumes that the CLD and the MEC apical plasmalemma interact in some way which causes the CLD to rise out of the cytoplasm enveloped in the plasmalemma. Here I present the evidence for the two possibilities. The first scenario is favoured, but the second cannot be ruled out.

We investigated the relationship between dam's pelvic and calf's dimensions with dystocia due to fetopelvic disproportion in the Holstein breed and estimated risk factors and dystocia probability. For this purpose, external pelvic measurements were performed in 402 heifers 15 ± 11 (1–38) days ante-partum and specific conformation measurements were obtained from their calves 1.7 ± 1.2 post-partum. Dystocia was defined as the inability of the heifer to complete parturition spontaneously within 120 min after the appearance of the amnion with normal presentation, position and posture or as having definite obstetrical obstacles within 60 min. Overall and fetopelvic disproportion dystocia incidence was 10.4% and 5.2%, respectively. Heifer measurements mainly influenced overall dystocia, whereas calf conformation was related solely with fetopelvic dystocia. Specifically, heifers with a small pelvis (hip width <49.95 cm, pelvic inlet area <333.2 cm2, pelvic volume <7799.2 cm3) had 2.8 to 3.5 times greater incidence of overall dystocia (19.0–20.8%) compared to heifers with a larger pelvis (incidence of 7.0–7.6%). Regarding calf factors, sex (male calves), body weight, chest circumference and fetlock joint circumference significantly increased the odds of experiencing dystocia due to fetopelvic disproportion compared with female, lighter or smaller calves. In a backward elimination model with independent variables treated as continuous, an area under the ROC curve of 0.66 regarding the prediction of overall dystocia based on heifer pelvic length, and of 0.64 for the prediction of fetopelvic dystocia based on fetlock joint circumference was found. The combination of the two variables in one model improved the ROC area to 0.71 regarding dystocia due to fetopelvic disproportion, reaching acceptable level of discrimination. Our findings indicate that dystocia due to fetopelvic disproportion in heifers is mainly influenced by the fetal side. Additionally, the estimation of pelvic dimensions of the dam before parturition and specific conformation characteristics of the calf during parturition, especially fetlock joint circumference, could aid obstetricians and herdsmen regarding dystocia probability and parturition surveillance.

This research communication describes a pilot study measuring skin carotenoid levels of lactating dairy cows. Carotenoids are natural antioxidants, involved in cell communication and immune function, protecting against oxidative stress. They are precursors of vitamin A, important for reproduction efficiency, growth and male fertility. Therefore, easy-to-use, inexpensive methods to measure carotenoids in cattle would provide interesting data for farmers to monitor the health and nutritional status of their herds. In this study, we used a commercially available sensor based on multiple spatially resolved reflection spectroscopy (MSRRS), intended for human use, to measure the carotenoid content in bovine skin in three research herds in France, Ireland and Scotland. Carotenoid levels were measured by applying the sensor to the teat barrel, avoiding pigmented areas of skin. Mean sensor values differed significantly between herds and between diets, with pasture-based animals showing significantly higher carotenoid levels. Our results suggest that MSRRS can be used to accurately to measure skin carotenoids in cows. However, further calibration in bovines is needed to improve of the accuracy of the MSRRS sensor in cattle.

The starting objective of this research communication was to determine the prevalence of subclinical mastitis in buffalo in Turkey. We also seeked to isolate and identify staphylococci, determine their antimicrobial susceptibilities and biofilm-forming abilities as well as investigating the presence of biofilm-related genes and microbial surface components recognizing adhesive matrix molecules. A total of 107 (66.9%) staphylococci (28 S. aureus and 79 coagulase-negative staphylococci, CoNS) were isolated from 160 mastitic milk samples collected from 200 lactating water buffalos. The staphylococci were especially resistant to beta-lactams except for cefoxitin but were less resistant to the other antimicrobials that were tested. Based on the Congo red agar method, 92.9% of the S. aureus and 70.9% of the CoNS isolates were positive for biofilm-forming ability, while all S. aureus and 97.5% of CoNS isolates were positive by a microtiter plate analysis. The presence of icaA and icaD genes was not always correlated with biofilm synthesis, and even in the absence of these genes, the isolates were able to synthesize biofilm.

The milk fat globule membrane (MFGM) is formed by complex cell biological processes in the lactating mammary epithelial cell which result in the release of the milk fat globule (MFG) into the secretory alveolus. The MFG is bounded by a continuous unit membrane (UM), separated from the MFG lipid by a thin layer of cytoplasm. This unique apocrine secretion process has been shown in all of the mammary species so far investigated. Once the MFG is released into the alveolus there is a considerable transformation of the UM with its attached cytoplasm. This is the MFGM. The transformation is stable and expressed milk shows the same transformed MFGM structure. Again, this transformation of structure is common to all mammalian species so far investigated. However, the explanation of the transformation very much depends on the method of investigation. Transmission electron microscope (TEM) studies suggest a literal breakdown to a discontinuous UM plus cytoplasm in patches and strands, whereas more recent confocal laser scanning light microscopy (CLSM) studies indicate a separation, in a continuous UM, of two phases, one liquid ordered and the other liquid disordered. This review is designed to show that the TEM and CLSM results show different views of the same structures once certain deficiencies in techniques are factored in.

We carried out a thorough genetic evaluation of Streptococcus dysgalactiae isolated from clinical bovine mastitis cases and performed a phylogenetic analysis to represent the evolutionary relationship between S. dysgalactiae sequences. A total of 35 S. dysgalactiae strains were isolated from cases of clinical mastitis identified at a large commercial dairy farm located near Ithaca, New York. Whole-genome sequencing identified twenty-six antibiotic resistance genes, four of which were acquired genes, in addition to fifty virulence genes. Multi-locus sequence typing detected three new sequence types (STs). We conclude that a high proportion of this microorganism carries multiple virulence determinants and resistance genes, and that this indicates its potential to cause mastitis. Eight different STs were identified, of which ST453 (n = 17) was the most prevalent and ST714, ST715, ST716 were novel STs.

To contribute to the treatment decision and optimize coagulase-negative staphylococcus (CNS) control programs, we conducted a meta-analysis to investigate the epidemiology and antimicrobial resistance rates of coagulase-negative staphylococcus associated with bovine mastitis in China. Three databases (PubMed, Google scholar and China National Knowledge Infrastructure database) were utilized to obtain relevant publications. A total of 18 publications were included in our research, and 3 of them included antimicrobial resistant (AMR) test. The pooled prevalence of coagulase-negative staphylococcus was 17.28%. Subgroup analysis revealed that the prevalence was higher in South China than in North China, was higher in 2011–2020 than in 2000–2010 and was higher in clinical bovine mastitis cases than in subclinical cases. The pooled AMR were most resistant to β-lactams, followed by tetracyclines, quinolones, nitrofurans, lincosamides, sulfonamides, amphenicol and aminoglycosides. The pooled AMR rate of coagulase-negative staphylococcus was lower in 2011–2020 than in 2000–2010. Although the prevalence of CNS showed an increasing trend over 20 years, the AMR rate showed a decreasing trend, and the clinical type of mastitis was the most frequent and the prevalence was highest in South China. Finally, CNS was most resistant to β-lactams amongst the eight groups of antimicrobial agents.

This research paper presents the development and evaluation of pioneering nanocomposites (NCs) based on the combination of k-carrageenan and linseed mucilage. When loaded with macela extract nanoemulsion they present an innovative approach for the sustained release of antimicrobial herbal constituents, specifically tailored for bovine mastitis treatment. The NCs, encompassing various ratios of k-carrageenan and linseed mucilage polymers (8:2, 7:3, and 5:5 w/w) with 1.25 mg of macela extract/g of gel, underwent in vitro assessment, emphasizing viscosity, degradation speed, release of herbal actives from macela nanoemulsion and antimicrobial activity. The NCs exhibited thermoreversible characteristics, transitioning from liquid at 60°C to a gel at 25°C. NCs allowed a gradual release of phenolic compounds, reaching approximately 80% of total phenolics release (w/v) within 72 h. NCs inhibited the growth of MRSA (ATCC 33592) until 8 h of incubation. No toxic effect in vitro of NCs was found on MAC-T cells. Thus, the developed materials are relevant for the treatment of bovine mastitis, especially in the dry period, and the data support future evaluations in vivo.

This research paper addresses the problem that, thus far, there is no method available to predict herd resilience for farms that do not use automated milking systems (AMS). Recently, a methodology was developed to estimate both individual cow as well as herd resilience using daily milk yield observations at individual cow level from farms with AMS. This AMS-based method, however, is not suitable on farms that use conventional milking systems (CMS) where such individual cow milk yield observations are lacking. Therefore, this research aimed at predicting herd resilience using herd performance data that is commonly available on CMS farms. To do so, data consisting of 585 Dutch AMS farms where herd resilience estimates using the AMS-based method were available was examined. To predict herd resilience with herd performance data, only those data that are also commonly available on CMS farms were used in a 5-fold cross validation Random Forest model. These herd resilience estimates were subsequently compared with the AMS-based herd resilience estimates. Results showed that it is possible to predict with a 69.9% probability whether a herd performs with above or below average herd resilience using only variables available on CMS farms. Especially, the proportion of cows with an indication of rumen acidosis, proportion of cows with an elevated somatic cell count and the fluctuation in herd size over the years are good predictors of herd resilience. Since herd management decisions appear to affect herd resilience, a lower predicted herd resilience could be taken as a general indication that tactical or strategic management changes could be taken to improve the herd resilience.

The research described in this Research Communication addresses the hypothesis that intramammary infections with Streptococcus uberis (S. uberis) are associated with biofilm formation, which limits antibiotic efficacy. This retrospective study investigated biofilm expression and antimicrobial resistance (AMR) patterns of 172 S. uberis infections. Isolates were recovered from milk samples of subclinical, clinical, and intramammary infection cases on 30 commercial dairy herds. We determined the presence and intensity of biofilm expression of S. uberis isolates in vitro in three somatic cell count categories to recognise their AMR patterns. An automated minimum inhibitory concentration system with a commercially available panel of 23 antimicrobial agents evaluated AMR, while biofilm determination was conducted using a microplate method. The study established that all the S. uberis isolates assessed expressed biofilm with the following varying degrees of intensities: 30 (17.8%) strong, 59 (34.9%) medium and 80 (47.3%) weak biofilms. The newly registered UBAC mastitis vaccine containing biofilm adhesion components may, therefore, be a viable option for proactive mastitis management under field conditions. No differences were identified between biofilm intensity and the three somatic cell count groups. Most S. uberis isolates indicated a high-level sensitivity to the antimicrobial agents tested. Resistances were present in 8.7, 8.1 and 7.0% cases to rifampin, minocycline and tetracycline, respectively. Multidrug resistance was observed in 6.4%, emphasising AMR to antibiotics used in human medicine only. The low overall resistance suggests that farmers adhere to the prudent use of antimicrobials in the dairy industry.