The study presented in this Research Communication aimed to investigate the relationship between physiological responses, body surface temperature and shade-seeking behaviour in Nili Ravi dairy buffaloes during summer months. We enrolled 60 buffaloes, and each animal was observed for three consecutive days starting before sunrise until they moved towards the shade structures. A repeated measures ANOVA was employed to assess the changes in physiological parameters and body surface temperature between the early morning and the occurrence of shade-seeking behaviour. The average temperature humidity index and heat load index during the behavioural monitoring period (0400 to 1200 h) were 81.3 ± 6.5 and 92.9 ± 17, respectively (mean ± sd). There was no significant difference in core body temperature between sunrise and the time of shade-seeking event. However, the buffaloes had a slightly higher respiration rate at the time of shade-seeking (19.2 vs. 22.4 breaths/min). In addition, body surface temperature, measured at the flank region, shoulder, base of the ear and forehead was significantly higher at the occurrence of shade-seeking behaviour compared to the early morning. On average, the buffaloes sought shade when the surface temperature was 2°C higher than the temperature recorded before sunrise. Overall, the current findings suggest that body surface temperature, rather than core body temperature was strongly associated with shade-seeking behaviour in dairy buffaloes. These findings could be useful in developing strategies to mitigate the effects of heat stress in dairy buffalo herds and thereby improve animal welfare.

Rapid diagnostic tests that differentiate between Gram positive, Gram negative and the absence of aerobic bacteria in milk samples from dairy cows with clinical mastitis can support antimicrobial treatment decisions and contribute to a more prudent use of antimicrobials in the dairy industry. The objective of this study was to evaluate the test characteristics of the novel rapid BACT mastitis test in discriminating causes of clinical mastitis under laboratory conditions. Test outcomes of 155 milk samples from clinical mastitis cases were incubated for 14–16 h in the BACT test and compared to results of bacteriological culture. The accuracy for detection of bacterial growth and Gram positive growth was 91 and 89%, respectively. The BACT test could provide an accurate and relatively fast decision tool for farmers to aid in antimicrobial treatment decisions in cases of clinical mastitis.

We investigated the antimicrobial components in cow milk at dry off and postpartum and their contribution in preventing new high SCC at quarter level. Milk samples from 72 quarters of 19 lactating cows were collected at last milking before dry off and at 7 d after parturition. Milk yield of each cow was recorded and SCC, IgG, IgA, lactoferrin, lingual antimicrobial peptide (LAP), and S100A7 concentrations in each quarter milk sample were measured. The postpartum milk yield was significantly higher than that at dry off. The IgG, IgA and lactoferrin concentrations in milk at dry off were significantly higher than those at postpartum, whereas the LAP concentration was lower. Quarters with SCC < 300 000 cells/ml at both dry off and postpartum were classified as persistent low SCC (PL) whereas those that rose above that same threshold postpartum were classified as new high SCC (NH). At dry off, IgG and LAP concentrations in milk were significantly higher in PL than in NH. These results suggest that high LAP concentrations during the dry period may contribute toward the prevention of new high SCC.

This Research Communication introduces a novel enzymatic-fluorometric analytical procedure for glycerol and glycerol 3-phosphate in milk. Milk from thirty-seven goats was analysed during 9 consecutive days during which a two-day feed restriction was introduced. Fractional milk triacylglyceride and free glycerol increased significantly while glycerol 3-phosphate reacted more moderately. The energy status of the mammary cell is discussed.

One of the major challenges in lameness management is prompt detection, especially before visible gait disturbance. This scoping review describes the potential biomarkers for lameness in dairy cows reported in the literature, their relevance in lameness diagnosis, identifying cows at risk of developing claw lesions and monitoring recovery after treatment. Using specific keywords, a comprehensive literature search was performed in three databases: PubMed, Google Scholar and ScienceDirect to retrieve relevant articles published between 2010 and 2022. A total of 31 articles fulfilling the inclusion criteria were analysed. The categories of potential markers for lameness reported in the literature included acute phase proteins (APPs), nociceptive neuropeptides, stress hormones, proteomes, inflammatory cytokines and metabolites in serum, urine and milk. Cortisol, APPs (serum amyloid A and haptoglobin) and serum, urinary and milk metabolites were the most studied biomarkers for lameness in dairy cows. While APPs, nociceptive neuropeptides and blood cortisol analyses assisted in elucidating the pain and stress experienced by lame cows during diagnosis and after treatment, evidence-based data are lacking to support their use in identifying susceptible animals. Meanwhile, metabolomic techniques revealed promising results in assessing metabolic alterations occurring before, during and after lameness onset. Several metabolites in serum, urinary and milk were reported that could be used to identify susceptible cows even before the onset of clinical signs. Nevertheless, further research is required employing metabolomic techniques to advance our knowledge of claw horn lesions and the discovery of novel biomarkers for identifying susceptible cows. The applicability of these biomarkers is challenging, particularly in the field, as they often require invasive procedures.

The study was carried out in dairy cows to elucidate whether treatment of clinical mastitis quarters with Spectramast® LC (ceftiofur hydrochloride, 125 mg, Zoetis) created a reason for discarding milk from adjacent untreated healthy quarters. The antibiotic was infused once daily in the affected mammary quarter for four days. Forty-nine cows were evaluated after diagnosis of clinical mastitis in three or fewer udder quarters. In all cases, quarters that did not receive treatment had milk samples collected one day after the end of treatment. All milk samples from untreated quarters were below the maximum permissible limit for the presence of antibiotic residues after analysis with the BetaStar S Combo test. Pharmacokinetic and pharmacodynamic characteristics may explain this finding. We conclude that it is feasible to use milk from untreated quarters of animals that have been treated with Spectramast® LC. We also reiterate the need to carry out tests with other pharmacological bases, and that the results found in this experiment cannot be extrapolated to other drugs.

Dairy cattle have considerable importance in the development of the Brazilian economy, being directly linked to economic and social progress. In the first half of 2020, 12.1 billion liters of milk were produced in Brazil and in 2019, there was a new record of 25.01 billion liters produced (IBGE, 2020). This production comes from a wide variety of production systems, coming from smallholder farmers as well as from large companies that use the latest technologies available on the market. Dairy production is a complex activity. For one to obtain economical success, several aspects must be monitored. Maintaining the health of animals is a top priority, and the literature suggests that various diseases are a common challenge for dairy producers. Mastitis is the main disease that affects dairy cows, responsible for considerable economic loss and significant zootechnical and productive challenges (Ruegg, 2017). It is considered the second leading cause of cow culling in dairy herds, behind reproductive problems. Mastitis is characterized by infection of the mammary gland and may or may not occur with inflammation, generating changes in the mammary tissue and properties of the milk. It is classifield into clinical or subclinical mastitis, according to presence or absence of clinical signs, and into contagious or environmental based on the causative agent (Correa et al., 2001).

The aim of this study was to compare the effects of feeding pasteurized waste milk or saleable milk to calves on weight, health and emergence of antimicrobial resistance in Escherichia coli strains isolated from those calves. An experimental study under field conditions on a commercial pasture-based Argentinian dairy farm was carried out. Forty Holstein calves were assigned randomly to either pasteurized waste milk (PWM) or non-pasteurized saleable milk (SM). The antimicrobial agents (AM) used on the farm, both to treat or prevent diseases, were recorded. The passive immunity level, calf live weight, AM presence in milk, clinical examination of calves, and E. coli isolation and identification, were performed. A total of 258 E. coli strains were isolated from fecal samples (132 isolates from SM calves and 126 from PWM calves at six sampling times). All E. coli isolated were used to perform AM susceptibility tests (disc diffusion and agar dilution). No differences were observed between groups in health parameters, average daily gain or prevalence of resistant E. coli strains to any AM evaluated throughout the study. Peaks of trimethoprim, sulfamethoxazole and enrofloxacin minimum inhibitory concentration (MIC) were observed at 30 d in E. coli from both groups of calves, whilst additional peaks to tetracyclin and ampicillin were observed only in SM calves. All MIC apart from gentamicin decreased at 75 and 90 d of age (during the weaning period). Gentamicin MIC behaved differently, having no peaks and increasing at 90 d only in PWM group. In conclusion, we found no evidence that emergence of antibiotic resistance is related to the consumption of pasteurized waste milk.

In this research communication we investigate the prevalence and antimicrobial susceptibility of S. aureus harboring virulent genes responsible for mastitis in cattle of Punjab, Pakistan. A total of 690 milk samples were collected from commercial dairy farms for analysis of the prevalence of subclinical and clinical mastitis and isolation of S. aureus. Virulence ability and methicillin resistance in S. aureus (MRSA) was determined by targeting the pvl (the gene for Panton–Valentine leukocidin) and mecA genes, respectively. A total of 175 S. aureus isolates exhibiting prevalence of pvl gene (6.28%) and mecA gene (22.28%) were determined. Antimicrobial susceptibility testing of pvl positive and negative MRSA against different classes of antibiotics revealed 100% resistance against β-lactams while 100% sensitivity towards tylosin and linezolid.

This research article describes an investigation into the udder health, bacterial isolation and antimicrobial sensitivity of three staphylococcal species isolated from the milk of non-dairy goats, suckling their kids, on two smallholder farms in the Hong Kong Special Administrative Region. Udder lesions were visually noted in 21 of 34 goats and two goats had palpable abnormalities. Collected milk samples grew a total of 11 bacterial organisms and the most frequently isolated organism was Staphylococcus chromogenes. Selected isolates of S. aureus, caprae and simulans from both farms were tested by antimicrobial sensitivity testing for 23 antimicrobials and all isolates showed antimicrobial resistance to doxycycline and tetracycline. Less common resistance was shown to ampicillin, chloramphenicol, penicillin and rifampicin. This preliminary study confirms the presence of udder lesions and mastitis bacteria in non-dairy goats in Hong Kong, along with the first information on the antimicrobial profile of three common Staphylococcus species bacteria affecting goats.

In this research paper the in vitro antimicrobial and antibiofilm activity of phage cocktail against the coagulase negative Mammaliicoccus sciuri was investigated. Three M. sciuri isolates obtained from clinical bovine mastitis samples were characterized and identified by 16S rRNA gene sequencing. Bacteriophages with lytic activity against M. sciuri isolates were isolated from dairy farm effluents. Two typical phages were isolated using standard enrichment and plaque assay techniques, purified by polyethylene glycol precipitation, and morphologically characterized based on shape and size using transmission electron microscopy. This was followed by determination of host range using spot tests and stability to varying temperature, pH and UV treatment. The phage cocktail suppressed bacterial activity within 30 min of exposure. Crystal violet assay showed that the tested phages and their cocktail significantly reduced the biofilm biomass of all three M. sciuri strains compared to the untreated control in vitro within 24 h with a single dosing. Transmission electron micrography of the purified phage particle revealed an icosahedral head and a rigid contractile tail, characteristic of the class Caudoviricetes. The findings open new avenues in phage-based antimicrobial approaches for controlling contagious and teat skin opportunistic bacteria causing bovine mastitis.

The protein composition in goat milk undergoes changes throughout the different lactation periods, displaying distinct characteristics that are influenced by the dynamic nature of protein composition and concentration during the transition from colostrum secretion to mature milk. To evaluate the dynamics of whey proteins of Saanen goats during the colostral phase and the first month of lactation, 110 milk samples from 11 healthy mammary halves of seven Saanen goats were selected through a clinical evaluation. Whey was obtained by rennet coagulation of the mammary secretion. The biuret method determined total protein concentration, and their fractions were identified by 12% dodecyl sulfate-polyacrylamide gel electrophoresis. Maximum concentrations of all protein fractions were observed in the first 12 h of lactation, reducing throughout the study. Modification of the protein predominance was also observed. The transition from colostrum secretion to milk occurred 5 or 7 d postpartum.

The geometry of milk liners may affect milking performance and cow comfort as the milk liner is the only part of the milking machine that comes into contact with the teat. To determine the effect of alternative shape of milk liners we compared square (SQR) vs. the conventional round (RND) teat cup liner on milking performance and comfort of dairy cows. Treatment milk liners were randomly allocated to clusters within each side of the 12 a side double up-herringbone dairy shed in a complete randomised block design over two periods. Milking performance data from a total of 10 065 (late stage of lactation and once-a-day milking frequency, LATE) and 18 048 (early stage of lactation and twice-a-day milking frequency, EARLY) milking events were automatically recorded by a DeLaval milk meter, and separately analysed for LATE and EARLY, respectively. In EARLY, cow comfort behaviour was also recorded during afternoon milking sessions. Across the two study periods, average milk flow rate, milk flow rate during 0–15, 15–30 and 30–60 s after cluster attachment, and milk flow rate at cluster take-off were higher in SQR compared to RND treatment. Proportion of time in a milking session with low milk flow rate and duration of milking session were less in SQR compared to RND treatment. However, effect of geometry of milk liner on peak milk flow rate was inconsistent across the two-study periods. Peak milk flow rate was higher (P < 0.001) in SQR than RND in LATE, but higher (P < 0.001) in RND than SQR in EARLY. Stomping and kicking behaviours of cows were similar between treatments. Results of this study suggest that square milk liners potentially improve milking performance, without adverse effect on cow comfort compared to conventional round liners. Long-term, multi-site studies are required to confirm potential teat-end health benefits associated with square milk liners and further verify these results.

This research paper analyzes the stability of raw cow milk in the alcohol test and seeks to understand to know the factors that influence milk stability and the occurrence of unstable non-acid milk. Milk samples were collected from the cooling tanks of rural farmers in the state of Paraná twice in summer and twice in winter. The farms were classified according to the production system: pasture with supplementation and feedlot. The following variables were analyzed: stability in the alcohol test, titratable acidity, ionized calcium concentration (iCa), chemical composition of milk, somatic cell count and standard plate count. The results showed that milk stability was greater in winter vs. summer, when the milk contained higher iCa, and in the feedlot vs. pasture system. The Pearson Correlation between variables (ethanol stability, milk composition, iCa, cooling tank temperature, milk volume, number of milking, number of cows milked, fat/protein ratio, distance and travel time) were analyzed. Stability was negatively correlated with iCa concentration and positively with lactose content. Logistic regression of the risk of unstable non-acid milk at 72% alcohol (UNAM72) showed that only iCa and lactose were determinants, while evaluation of the same risk at 78% alcohol revealed iCa, titratable acidity, lactose and milk urea nitrogen as risk factors. Under the dairy farming conditions of Paraná state, the frequency of UNAM72 was low (12.16%) and was higher in summer and in pasture systems with supplementation. In conclusion, in dairy herds bred with high technological level, with adequate nutritional and health management, the frequency of UNAM is low and is related to nutritional management abnd, perhaps, heat stress, factors that alter iCa and lactose levels.

We tested the hypothesis that milk proteins, through microencapsulation, guarantee protection against bioactive substances in coffee silverskin extracts. Therefore, the aim of this study was to carry out technological, nutritional and physicochemical characterisation of a coffee silverskin extract microencapsulated using instant skim milk powder and whey protein concentrate as wall materials. The aqueous extract of coffee silverskin was spray-dried using 10% (w/v) skim milk powder and whey protein concentrate. The samples were characterised by determining the water content, water activity, particle size distribution, colour analysis and total phenolic compound content as well as antioxidant activity using 2,2-diphenyl-radical 1-picrylhydrazyl scavenging methods, nitric oxide radical inhibition and morphological analysis. The product showed water activity within a range that ensured greater stability, and the reduced degradation of the dried coffee silverskin extract with whey protein concentrate resulted in better rehydration ability. The luminosity parameter was higher and the browning index was lower for the encapsulated samples than for the pure coffee silverskin extract. The phenolic compound content (29.23 ± 8.39 and 34.00 ± 8.38 mg gallic acid equivalents/g for the coffee silverskin extract using skimmed milk powder and whey protein concentrate, respectively) and the antioxidant activity of the new product confirmed its potential as a natural source of antioxidant phenolic compounds. We conclude that the dairy matrices associated with spray drying preserved the bioactive and antioxidant activities of coffee silverskin extracts.

The objective was to examine the prevalence of extended-spectrum β-lactam (ESBL) resistance among Shiga toxigenic Escherichia coli (STEC) isolated from raw milk. The findings from disc diffusion analysis and polymerase chain reaction revealed a high occurrence of ESBL resistance, specifically to penicillins, cephalosporins, and carbapenems, in isolated STEC strains. Additionally, a distinct sequence type of STEC was also identified in milk through comparative analysis.

The experiments reported in this research paper aimed to evaluate the physico-chemical and sensory characteristics, microbial quality and antioxidant potential of goat's milk paneer during storage (0–12 d, 4 ± 1°C). The juices from five different citrus fruits were used as coagulant (treatments) to make goat's milk paneer. The pH of all paneer samples decreased during storage whereas titratable acidity increased. Ash (%) fat (%) and protein (%) of paneer increased slightly during storage, whereas sensory perception decreased. The juices from all the citrus fruit varieties showed high contents of total phenolics and total flavonoids which ultimately influenced ferric reducing antioxidant power, total antioxidant capacity and radical scavenging activities. As the contents of different juices were also retained in the paneer matrix, so paneer coagulated with citrus juices also showed encouraging results in terms of total phenolic and flavonoid contents, ferric reducing antioxidant power and radical scavenging activities. Amongst all the paneers, the most promising was that coagulated by kinnow juice. In addition, the whey obtained from paneer coagulated by citrus juices also showed appreciable quantities of total phenolic and flavonoid contents, thereby beneficially influencing ferric reducing antioxidant power andradical scavenging activities. It is concluded that citrus juices improve the sensorial quality and antioxidant potential of goat's milk paneer and its whey.

We address the hypothesis that at early and late lactation milk presents low ethanol stability due to high acidity and ionic calcium values. Our aim was to evaluate the functional traits of milk (milk ethanol stability: MES, acidity and ionic calcium: iCa) according to lactation stage in different genetic groups. Raw milk samples were collected from Jersey (n = 271), Holstein (n = 248) and Jersey × Holstein crossbred cows (n = 82), raised on five commercial farms located in the state of Paraná, Brazil. Milk composition, somatic cell count (SCC), milk urea nitrogen (MUN), MES, pH, acidity and iCa were determined. Days in milk (DIM) were categorized into four classes: 1–60, 61–150, 151–305 and over 305 DIM. Data were submitted to analysis of variance. Fixed and random effects were incorporated into the model, in a repeated measures in time arrangement using the mixed models methodology. Significant interactions between DIM class and genetic groups were detected. The comparison between each combination of genetic group and DIM class showed that at the beginning of lactation, Holsteins produced milk with higher MES than Jersey and crossbreds. At 105–305 DIM Holstein milk presented higher MES than Jersey, while beyond 305 DIM Holstein milk showed higher MES than crossbred cows. At the beginning of lactation acidity was higher in Holstein milk and crossbreds compared with Jersey, while acidity was lower in Holstein milk compared with Jersey and crossbreds in the other lactation stages. Ionic calcium was highest after lactation peak for Holstein, but did not vary between lactation stages for Jersey and crossbreds. Functional characteristics of bovine raw milk such as MES, iCa and acidity varied between lactation stages in a distinct manner according to genetic groups. Early and end lactation stages are challenging in terms of low stability, especially for Jersey and crossbreds.

α-Lactalbumin (α-LA) and β-lactoglobulin (β-LG) are major whey proteins in bovine milk. We studied the effects of these molecules on the intestinal cell response by comparing the native form with the denatured form containing oligomers obtained by treatment with 2,2,2-trifluoroethanol (TFE). We previously reported that proteins in native and TFE-treated forms exhibited cell growth stimulation and cytotoxicity, respectively, in undifferentiated rat crypt IEC-6 and human colon Caco-2 cells. However, neither whey protein showed cytotoxicity even in the TFE-treated form in differentiated Caco-2 cells. Only undifferentiated immature intestinal cells can distinguish between these native and denatured proteins. Moreover, α-LA and β-LG exhibited different oligomer formation characteristics during the TFE treatment. In the present study, we compared the effects of native and TFE-treated whey proteins on IEC-6 cells in more detail. The native forms of both whey proteins exhibited cell proliferative effects in a concentration-dependent manner. For the TFE-treated forms, α-LA showed rapid and potent cytotoxicity, whereas β-LG altered cell responses depending on its concentration and exposure time; lower concentration/shorter exposure and higher concentration/longer exposure induced cell growth stimulation and cytotoxicity, respectively. Pre-treatment of the cell membrane with cholesterol suppressed the effects on the cell response only in TFE-treated β-LG (TFE-β-LG). In a preliminary examination using inhibitors of signal transduction, TFE-treated α-LA acted on the intrinsic apoptosis pathway via Bcl-2-associated X and p53, whereas the action of TFE-LG did not require this pathway. Tyrosine phosphorylation is necessary for the cell proliferation effect of both native whey proteins; however, native α-LA, but not native β-LG, also required activation of the pathway with selective epidermal growth factor receptor tyrosine kinase and Janus kinase 2/3. In summary, the two major bovine milk whey proteins induced similar yet discrete responses in undifferentiated intestinal cells. Even when oligomers are formed, β-LG may be much less hazardous to immature intestinal cells than α-LA.

This research paper addresses the hypotheses that Kluyveromyces marxianus can be cultured with good alcohol production on different whey-derived matrices, and that the fermented product can be used in order to develop alcoholic beverages with acceptable sensory characteristics by mixtures with yeast-fermented fruit-based matrices. Growth and fermentative characteristics of Kluyveromyces marxianus LFIQK1 in different whey-derived matrices were explored by culturing (24 h, 30°C) on reconstituted whey, demineralized whey, heat-treated whey and milk permeate media. High lactose consumption, ethanol production and yield were observed. Reconstituted whey matrix was selected for mixing with orange or strawberry juices fermented using Saccharomyces cerevisiae to obtain alcoholic beverages (W-OR and W-ST, respectively). Consumer evaluation of beverages was performed using acceptability and Check-All-That-Apply (CATA) questions. Good acceptance was observed, significantly higher for W-ST than for W-OR. CATA questions gave information about organoleptic characteristics of beverages. Penalty analysis showed W-R and W-ST were positively associated with smooth/refreshing and fruity/natural, respectively. Liking was represented, accordingly with penalty analysis, by natural/refreshing. A novel alternative for utilization of whey and whey-related matrices by alcoholic beverages production with natural ingredients is presented.