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Background: Intramammary infections negatively affect milk quality, animal welfare and productivity in the dairy industry. Somatic cell count (SCC) is the most used screening tool to detect subclinical mastitis caused by intramammary infections. In dairy goats, SCC is greatly influenced by non-infectious factors, which complicates the interpretation. The aim of this research paper was to determine the association between SCC, intramammary infections and non-infectious factors including parity, season, lactation stage, and milk yield in dairy goats. In this longitudinal study, 451 goats from four Norwegian dairy goat herds were sampled for bacteriology and SCC up to nine times during two lactations. Factors like parity, milk yield, and stage of lactation were retrieved from the Norwegian goat recording system., Results: The most prevalent udder pathogen findings were Staphylococcus caprae (6.8%), Staphylococcus warneri (6.3%), and Staphylococcus epidermidis (3.8%), all of which had a mild but significant impact on SCC. Staphylococcus aureus was detected in 3.6% of the udder halves and had a major effect on SCC. Parity, stage of lactation, season, and milk yield significantly influenced SCC., Conclusions: This study highlights that intramammary infections caused by Staphylococcus aureus, along with factors such as increasing parity and the seasonal effects of pasturing, significantly influence the SCC. Understanding these key contributors is essential for improving udder health management and improving milk quality in goat milk production., (© 2024. The Author(s).)

Mastitis is a global concern in the dairy sector, demanding innovative solutions for effective management for quality lifetime milk production. In this study, infrared thermography (IRT) as a non-invasive technology was integrated into routine farm activities for continuous health monitoring of animals. For 30 days, we systematically monitored the udder health status in 40 Sahiwal cows (160 quarters), employing IRT along with the California Mastitis Test (CMT). We also assessed somatic cell count (SCC), microbial identification, and milk quality parameters of representative samples. The thermal imaging data was analyzed, considering both backward propagation from the 0 th day to the -10th day and forward propagation from the 0 th day to the +10th day. Our findings revealed that on the 0 th day, the mean temperatures of the udder surface skin temperature (USST) and teat skin surface temperature (TSST) exhibited differences (p < 0.05) between the quarters affected by sub-clinical mastitis (SCM) and clinical mastitis (CM) in comparison to the healthy quarters, with the highest degree of difference observed. The observed temperature differences between CM and SCM quarters compared to healthy ranged from 1.8 to 3.62 °C and 0.98 to 3.23 °C for USST, and from 1.68 to 3.16 °C and 0.56 to 2.32 °C for TSST, respectively. Furthermore, our observations indicated that both udder and teat quarters responded differently to mastitis. A temperature rise of 1.37 °C in SCM quarters and 1.75 °C in CM quarters was observed between the -10th and -8th day relative to day 0, with the increase being more pronounced in the morning hours. Also, a notable temperature surge occurred during the -2nd and -1st days relative to the 0 th day. The log 10 SCC values and milk quality parameters significantly differed (p < 0.05) between mastitis-affected and healthy samples. In addition, Staphylococcus spp. was identified as the predominant mastitis-causing pathogen in the bacteriological identification conducted in this study. Therefore, IRT efficiently assesses the initiation point of udder infection in Sahiwal cows, aiding in effective udder health management., Competing Interests: Declaration of competing interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: T. K. Mohanty reports financial support was provided by Indian Council of Agricultural Research, New Delhi, India. If there are other authors, they declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

The CellCheck dry cow consult (DCC) was developed by the CellCheck Technical Working Group to enable farmers to engage with their nominated veterinarian to develop farm-specific selective dry cow therapy (SDCT) plans, where appropriate. This study evaluated the effect of the DCC on farmer decision-making around dry cow therapy, and the udder health effects of implementing SDCT, in study herds over the 2019 and 2020 dry periods. The DCC was a 3-h consult, delivered and funded as part of the Targeted Advisory Service on Animal Health. Herds that completed a DCC were invited to register for a dry cow review the following year. The combined dataset for analysis across both years comprised 439 herds and 25,357 cows. Herd size ranged from 25 to 800. The median SCC of cows dried off with teat sealant only was 47,000 cells/mL before drying off in 2019 and 48,000 cells/mL at first milk recording in 2020, and 43,000 cells/mL before drying off in 2020 and 39,000 cells/mL at first milking recording in 2021. Following both the 2019 and 2020 dry periods, cows tended to converge toward a similar median SCC early in the following lactation, irrespective of prior dry cow treatment strategy. The uptake of SDCT was greater in review herds, with 21% of cows receiving teat sealant only in 2020, compared with 16.3% of cows in herds participating in a consult for the first time in 2020. At an individual cow level, in both years dry period new infection rate (NIR) was approximately 2.7% higher for cows treated with teat sealant only, than for those treated with both dry cow antibiotic tubes and teat sealant, and 1.2% higher than cows treated with antibiotic only. Regardless of treatment, a significant association was detectable between increasing parity and the risk of a dry period new infection. Increasing herd size had a significant effect on the risk of dry period NIR. At a herd level, no significant increase in NIR occurred when SDCT was used compared with herds where blanket dry cow therapy was used. Though not without risk, SDCT can be successfully implemented in Irish herds; however, constant attention to hygiene and management is essential. Despite the challenges, facilitating continued farmer education and engagement with professional guidance will be important., (The Authors. Published by Elsevier Inc. on behalf of the American Dairy Science Association®. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).)

Background: Livestock farmers face financial losses every year because milk yield and components are severely affected by udder diseases. These udder infections attract the immune response from the host and lead to the influx of neutrophils into milk to fight infection and thus the number of somatic cell count (SCC) is increased. The SCC value of milk could be used as an important indicator in detecting clinical mastitis in dairy animals. Also, the milk yield and milk quality (e.g. fat) are negatively affected by the increased SCC. The SCC is used to estimate the somatic cell score (SCS)of the milk, which is used as an indirect measure to detect subclinical mastitis. Therefore, the purpose of this study was to investigate the presence of a significant SNP rs868996547, on the suppressor of cytokine signaling 2 gene (SOCS2) which is related to milk yield and milk quality in Awassi sheep., Methods: In this study, milk production data was obtained from 210 healthy Awassi ewes with different parties and ages. The general linear model (GLM) process analysis of variance (ANOVA) was used to determine fixed effects on milk traits. The DNA extraction was done using a blood DNA extraction kit from Qiagen. To validate the presence of SNP a customized SNP detection developed by Thermofisher Scientific was used. The presence of the SNP in the SOCS2 gene was detected with genotypes (C/T, T/T, and C/C) and T being the mutated allele and it had a significant (p < 0.015) effect on the milk yield (p < 0,015;0.091), fat (p < 0,001;0,003), fat/protein ratio (p < 0.001;0,037) and log10SCC value (p < 0,006;0,015) of Awassi ewes. However, the protein, total solid, and lactose percentages in the wild type and the mutated ewes found having no significant difference (P > 0.05)., Conclusion: Our result showed the increase in SCC or SCS of the milk significantly affected the milk yield and composition. Parity and age had significant effects on ewes' milk yield (p < 0.001). In conclusion, we investigated the presence of SOCS2 gene of Awassi ewes in the study flock and its effect on milk yield, fat, and somatic cell count, and the change in milk composition and milk yield because of SCC., (© 2024. The Author(s), under exclusive licence to Springer Nature B.V.)

The objective of this study was to determine quarters requiring antimicrobial treatment using either a benchtop somatic cell counter or culture with gram-positive selective media and compare the outcomes in these cows to those receiving blanket dry cow therapy (BDCT) in a randomized, controlled trial. We evaluated 2 novel methods of identifying cows with intramammary infections followed by selective antimicrobial treatment at a commercial dairy farm to determine their usefulness in decreasing antibiotic usage during the dry period without significant detrimental effects on milk quality and production. Cows (n = 840) were randomly allocated to one of 3 groups (BDCT, gram-positive selective media culture-based selective dry cow therapy [C-SDCT], and somatic cell count-based SDCT [S-SDCT]) the day before dry-off, and quarter-level milk samples (QLMS) were collected. The QLMS from cows in the S-SDCT group were evaluated using the cell counter, and quarters were treated if SCC was ≥200,000 cells/mL, whereas the QLMS from cows in the C-SDCT group were cultured, and quarters were treated if the culture showed growth. All cows in the BDCT received antimicrobial therapy, and all cows received an internal teat sealant regardless of treatment group. Outcomes measured were first and second DHIA test SCC, milk production through 60 DIM, cows leaving the farm, clinical mastitis, and bacteriologic new infections in a subset of quarters. Cows in both SDCT groups had fewer antimicrobial treatments than cows in the BDCT group as was expected, and cows in the C-SDCT group had fewer treatments than those in the S-SDCT group. Cows in both SDCT groups had a higher linear score at the first DHIA test (BDCT: 1.8, S-SDCT: 2.2, C-SDCT: 2.2); however, we found no other differences between groups regarding any other outcomes measured. Although antimicrobial use was significantly reduced, farms should use caution in adopting the benchtop analyzer and the selective media described in this study as ways to identify infected cows for dry cow therapy because they may result in increased linear scores early in lactation., (The Authors. Published by Elsevier Inc. on behalf of the American Dairy Science Association®. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).)

This observational study aimed to explore the association of farmer-driven selective dry cow therapy (DCT), milking routine and dry cow management practices with SCC in early-lactation cows from 21 commercial dairy herds. Milking routine practices evaluated referred to cow preparation for milking, in-lactation mastitis management, and recording. Dry cow management practices related to dry cow environment and cleaning, dry-off procedure, milk cessation strategy and calving environment. Data from 2,016 multiparous cows in 21 commercial spring-calving grazing dairy herds were available for the study. Herd owners self-reported DCT (the assignment and administration of DCT was at the discretion of the herd owners with no involvement from the research team), management practices during milking and the dry period. Cow-level last test-day SCC records in 2020 (range = 105-285 DIM) and first test-day records in 2021 (range = 5-60 DIM) were obtained from milk recording databases. Quarter-level milk sampling was carried out on all cows in late lactation of 2020 (range = 240-261 DIM) for bacterial culturing. Bacteriological results were used to define cows with IMI when ≥1 quarter sample resulted in bacterial growth and there were no contaminated samples from that cow. Mixed model analyses were used to evaluate the association of selective DCT, milking routine, and dry cow management practices with cows' first test-day log 10 SCC (logSCC) in 2021. At dry-off in 2020, 47.6% of the cows were administered an internal teat sealant alone (ITS), while 52.4% were administered an antibiotic plus an internal teat sealant (AB+ITS). The mean herd-level percentage of cows with IMI was 19.7% (range = 9.8%-39.5%); Staphylococcus aureus accounted for the majority of cow-level IMI (89.9%, 357/397). Between herds, the proportion of cows administered ITS ranged from 17.7% (14/79; in a herd with an IMI prevalence of 10.1%) to 86.8% (66/76; in a herd with an IMI prevalence of 27.6%). In total, 11.8% (105/889) and 29.8% (292/980) of cows that were administered ITS or AB+ITS had an IMI in late lactation 2020, respectively. The mean untransformed SCC at the last test day in 2020 of cows administered ITS and AB+ITS was 55,000 and 197,200 cells/mL, respectively. The statistical analysis showed a significant interaction between selective DCT and milk yield at last test-day in 2020; cows with a milk yield of 15 kg and administered ITS had a 0.1 higher (untransformed SCC of 19,000 cells/mL higher) first test-day logSCC compared with cows administered AB+ITS. Additionally, greater parity, IMI in late lactation, higher logSCC at the last test-day in 2020 and longer dry periods were associated with higher logSCC at the first test-day in 2021. The current study identified cow- and herd-level management practices that could aid dairy farmers in improving the outcome of selective DCT and decrease early lactation SCC., (The Authors. Published by Elsevier Inc. on behalf of the American Dairy Science Association®. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).)

The aim of the study was to investigate the effect of the immunostimulant Mycobacterium Cell Wall Fraction (MCWF) on the treatment of S. aureus SCM by intravenous application. The study included 45 HF dairy cows in 2nd and 3rd month after parturition divided into three groups (n = 15 per group): the MC + group - cows with S. aureus SCM treated with MCWF; the MC- group - cows with S. aureus SCM, with no treatment; and the C group - the control group of healthy cow with no treatment. Samples were collected 0th (I sample), 7th (II), and 14th day (III) from the day of SCM diagnosis and on day 21st (IV). A greater influx of leukocytes was confirmed into milk after 7 days after MCWF treatment in MC + group, which was followed by increase of WBC and LYM in blood. These results support the hypothesis of effective action of MCWF, and in quarters with lower-grade infection, bacteriological cure was achieved. The MC- group had a statistically higher concentration of TBARS and CAT activity in milk, while MC + group had lower blood serum LDH activity, which indicates a positive effect of the MCWF application and a lower exposure of the tissue to lipide peroxidation and inflammation caused by S. aureus. The application of MCWF would give new possibilities in the prevention and therapy of mammary gland diseases without fear of the presence of residues and the emergence of bacterial resistance. In future studies, the effects of local and systemic application of MCWF in the treatment of S. aureus SCM should be compared., (© 2024. The Author(s), under exclusive licence to Springer Nature B.V.)

This study aimed to monitor the mammary health of 37 multiparous Murrah buffaloes through infrared thermography (IRT). Based on the California Mastitis Test (CMT) and milk somatic cell counts (SCC), buffaloes were grouped into healthy (H, n = 16), subclinical mastitis (SCM, n = 10), and clinical mastitis (CM, n = 11). Buffaloes were milked twice daily in the morning (5:00-6:00 AM) and evening (5:00-6:00 PM). Rectal temperature and respiratory rates were recorded, CMT was performed and thermal images of the mammary gland of all the buffaloes were taken before and after each milking. Milk samples were analysed after each milking for SCC, fat, Solids-Not-Fat (SNF), density, protein, lactose, salts, conductivity, and pH immediately in the laboratory from fresh milk samples. The surface temperature of the periocular region of both the eyes, muzzle, flank, and vagina were also taken. Thermal images were used to assess the surface temperature of the udder (USST), teat apex (TAT), teat barrel (TB1T), teat base (TB2T), and teat skin surface (TSST). Eye and USST showed significantly higher temperatures (p < 0.05), whereas skin surface temperatures (SST) of different body parts were non-significant in both SCM and CM animals than buffaloes in the H group. Milk SCC showed a positive correlation with conductivity (r > 0.7), salts, and pH (r < 0.6) and a negative correlation with fat, SNF, density, protein, and lactose. TAT, TB1T, TB2T, TSST, and USST were positively correlated with milk SCC. Receiver Operating Characteristic (ROC) analysis of H and SCM groups showed that USST before milking had optimum sensitivity (Se = 0.80) and specificity (Sp = 0.906) among the various skin temperatures recorded. Thermal images captured during the morning showed higher sensitivity compared to images taken in the evening. Results indicate IRT can be used to monitor the mammary health of buffaloes but using IRT in conjunction with milk SCC can help in the accurate prediction of SCM in dairy buffaloes., (© 2024. The Author(s), under exclusive licence to Springer Nature B.V.)

Mastitis is a global production disease that needs an intelligent solution to tackle effectively. Infrared Thermography (IRT) is a non-invasive technology that could be incorporated into routine day-to-day farm activities to monitor the health status of the animals. In this study, the udder health status was routinely monitored for 30 days among 41 Murrah buffaloes via IRT and the California Mastitis Test (CMT). Further, somatic cell count (SCC), microbial identification, and milk quality parameters were also estimated for representative samples. The thermal imaging data obtained was tabulated and back propagated from the 0th day to the -10th day and front propagated from the 0th day to +10th day for all the udder quarters. Results revealed that on the 0th day, the mean of udder skin surface temperature (USST) and teat skin surface temperature (TSST) showed a difference (p < 0.05) in the sub-clinical mastitis (SCM) and clinical mastitis (CM) affected quarters to the healthy quarters, and their degree of difference was the highest. The indication of infection was signaled during the -9th to -5th day to th e 0th day in SCM and CM cases. There was a steep increment in the temperature from -2nd and -1st day to the 0th day of infection. Sometimes, some quarters show an increment in temperature due to mastitis during morning hours but recover by evening milking due to the animal's innate immune system. Thus, the initiation period in which the udder gets assaulted is crucial in the early assessment of SCM by monitoring temperature change using IRT., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Total bacterial count (TBC) and SCC are important quality parameters in goat milk. Exceeding the bulk milk TBC (BMTBC) thresholds leads to price penalties for Dutch dairy goat farmers. Controlling these milk quality parameters can be challenging, especially around kidding. First, we describe the variation and the peaks around kidding of TBC and SCC in census data on Dutch bulk milk over the last 22 yr. Second, to explore causes of these elevations, we studied the variation of TBC and SCC in individual goat milk from 3 wk before to 5 wk after kidding and their association with systemic response markers IFN-γ, calprotectin, BHB, BCS, and fecal consistency. We visited 4 Dutch dairy goat farms weekly for 10 to 16 wk around kidding. Some of the goats had been dried off; other goats were milked continuously throughout pregnancy. A total of 1,886 milk samples from 141 goats were collected for automated flow cytometric quantification of TBC and SCC measurement. IFN-γ, calprotectin, and BHB were determined twice in blood of the same goats; most samples were collected after kidding. The BCS and fecal consistency were scored visually before and after kidding. We found a strong correlation between TBC and SCC (Spearman's rho = 0.87) around kidding. Furthermore, in the third week before kidding, the average TBC (5.67 log 10 cfu/mL) and SCC (6.70 log 10 cells/mL) were significantly higher compared with the fifth week after kidding, where the average TBC decreased to 4.20 log 10 cfu/mL, and the average SCC decreased to 5.92 log 10 cells/mL. In multivariable linear regression models, farm and stage of lactation were significantly associated with TBC and SCC, but none of the systemic response markers correlated with TBC or SCC. In conclusion, TBC and SCC in dairy goats were high in late lactation and decreased shortly after parturition. For SCC, the dilution effect might have caused the decrease, but this was not plausible for TBC. Moreover, the excretion of bacteria and cells in goat milk was not associated with the selected systemic response markers that were chosen as a readout for general immunity status, intestinal health, and metabolic diseases. Therefore, we assume that the TBC increase before kidding and the decrease after parturition are caused by other systemic, possibly hormonal, processes. To reduce BMTBC and bulk milk SCC, it would be advisable to keep milk of goats with highest numbers of bacteria and cells in their milk out of the bulk milk during end lactation. Further studies are needed to investigate the effects of withholding this end-lactation milk from the bulk tank., (© 2024, The Authors. Published by Elsevier Inc. on behalf of the American Dairy Science Association®. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).)

Mastitis is one of the most serious diseases that threatens the health of dairy animals. The somatic cell count (SCC) in milk is widely used to monitor mastitis. This study aimed to reveal the diversity of microorganisms in buffalo milk with high somatic cell count (SCC ≥ 3 × 10 5 cells/mL, n = 30) and low somatic cell count (SCC ≤ 5 × 10 4 cells/mL, n = 10), and identify the dominant bacteria that cause mastitis in a local buffalo farm. We also investigated the potential method to treat bacterial mastitis. The V3-V4 region of 16 S rDNA was sequenced. Results showed that, compared to the milk with low SCC, the high SCC samples showed lower microbial diversity, but a high abundance of bacteria and operational taxonomic units (OTUs). By in vitro isolation and culture, Escherichia coli, Staphylococcus aureus, and Klebsiella pneumoniae were found to be the leading pathogens, which is consistent with the 16 S rDNA sequencing data. We further isolated 3 of the main pathogens and established a pathogen detection method based on ELISA. In addition, the antibacterial effects of 10 antimicrobials and 15 Chinese herbal extracts were also investigated. Results showed that the microbial has developed tolerance to several of the antimicrobials. While the water extracts of Chinese herbal medicine such as Galla Chinensis, Coptis chinensis Franch, Terminalia chebula Retz, and Sanguisorba officinalis L can effectively inhibit the growth of main pathogens. This study provides novel insight into the microbial diversity in buffalo milk and a reference for the prevention, diagnosis, and treatment of mastitis., (© 2024. The Author(s), under exclusive licence to Springer Nature B.V.)

Current research aims to generate an alternative model to classical methods in the determination of subclinical mastitis at 4 levels (healthy, suspicious, subclinical, and clinical). For this purpose, multilayer perceptron (MLP) artificial neural networks (ANN) was developed as test model. 5 variables from the physical properties of milk somatic cell count (SCC), electrical conductivity (EC), pH, density, and temperature at fore milking (TFM) were included in the model in the classification of mastitis. Model performance was validated on test data (%25) and compared with the multinomial logistic regression (MNLR). MLP model has shown a satisfactory performance with an accuracy of 95.14% and - 141 of AIC score better than the control model (MNLR) of 80.27% and - 133 AIC despite using higher number of parameters (104). Since the main problem is to diagnose subclinical mastitis, which does not cause any visible symptoms, it was important to distinguish between absolute subclinical (suspicious excluded positives) and absolute healthy (suspicious included positives) ones. Therefore, optimum cut-off threshold was evaluated for these two different scenarios with only variable SCC the gold standard indicator of subclinical mastitis and results were compared in the interpretation of model performance. The results show that the 5-variable MLP model exhibits a high sensitivity of 93.22% (AUC = 0.95 for healthy ones) at low cutoff thresholds as well. New studies should provide a better understanding by evaluating economics, sustainability, animal welfare and health aspects together to determine the optimal threshold value., Competing Interests: Declaration of competing interest The authors declare that there is no conflict of interest regarding the publication of this manuscript., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Introduction and Objective: Correlations between the number of milk somatic cells (SCC), the number of microorganisms, and the content of basic components of milk were studied on five farms (F1-F5) with cows of the same breed, but with different milking systems., Material and Methods: From each farm, 50 Holstein Friesien milk samples were collected once a month (250 samples/month; n=3,000) during March 2022 - February 2023. Samples from farms F1 and F5 were tested for fat, protein, lactose, no fat dry matter content (FTIR spectroscopy), for the SCC (Fossomatic 7), and for the differential cells (Vetscan DC-Q)., Results: The highest fat content was confirmed on farm F5 (3.85 ± 1.70%) and F4 (3.82 ± 0.21%) with automatic milking system (AMS). However, from the point of view of protein content, these farms showed slightly lower values (<0.05). F1 did not meet the minimum required amount for fat content (2.84 ± 0.81%) set by the legislation of the Slovakia. The comparison shows that there is not much difference in cell size between healthy cells and mastitis cells. The average size of healthy cells was approximately 8.77 ± 0.49 μm. In the monitored period, the average values determined were at the level of 292,000/mL (5.46 ± 0.72 log10 SCC) in cow milk samples, while for the rest of the year, the values remained at 256,000/mL (5.40 ± 0.80 log10 SCC). F1 was categorized as a positive farm with a high TLC (total milk leucocyte count) concentration (5.58 log10 cells/mL, 406.65 ± 53.80 × 10 3 cells/mL) and a predominant NEU fraction (61%). Farms F2, F4, and F5 were classified as negative farms (TLC was 4.70 ± 0.26 log10 cells/ml)., Conclusions: According to the results, the size of SCCs in healthy milk does not differ from SCCs found in mastitis milk. From the results, it can be concluded that the transition to the latest generation of robotic milking method can positively affect milk production and its quality.

This study evaluated the influence of milk production, number of lactations, and days in milk (DIM) on the quality and composition of milk from dairy cows housed in a compost barn (CB) system. The study was carried out using a six-year database, counting 31,268 observations from 2,037 cows of European breeds. Multiparous cows showed higher fat and protein production. Lactose showed high levels for primiparous and the initial stage of lactation (4.65%) and was negatively influenced by somatic cell count (SCC). Milk urea nitrogen was higher (14.01%) from 106 to 205 days in milk, and the other components were higher at >305 days. Therefore, the solids content was higher in the first and second lactations due to the high contents of lactose, fat, and milk protein, but lactose was reduced over lactations. In contrast, high DIM increased SCC and concentrated solids due to lower milk production. The effect of milk production, stage, and lactation order on the composition and milk quality of herds housed in CB showed the same pattern as in other production systems.

Mastitis, a multifactorial disease influenced by both cow and herd-level factors, results in significant losses throughout the dairy chain. We aimed to evaluate the relationship between milking frequency (MF), parity order (PO), days in milk (DIM), and milk yield (MY) on somatic cell count (SCC) and the odds of a cow having subclinical mastitis (SCM) in Brazilian Holstein and Jersey dairy cows. Our dataset consisted of 747,520 test-day records from 52,954 cows, including 49,089 Holstein cows and 3865 Jersey cows and 498 herds. The SCC was evaluated using a generalized linear mixed model, whereas SCM occurrence was evaluated using a logistic regression model. A case of SCM was defined when a cow had >200×10 3 cells/mL. Our results indicated that the SCC increases with higher PO and DIM and decreases in cows milked three times a day and those with higher MY in both breeds (>40 and >25 L/d for Holstein and Jersey, respectively). Increasing MF from two to three times a day reduced the chances of a Holstein and Jersey cow having SCM by 10 and 20 %, respectively. For Holstein and Jersey cows, those with ≥quadriparous had 3.9 times and 2.2 times higher chances, respectively, of having SCM compared to primiparous cows. Cows with >305 DIM had 2.0 times greater chances of having SCM for both, Holstein and Jersey cows, compared to cows with ≤105 DIM. Holstein cows yielding ≥40 L/d had a 75 % lower chance of having SCM compared to those yielding <20 L/d, while Jersey cows with ≥25 L/d had a 60 % lower chance compared to those yielding <15 L/d. In conclusion, higher PO and DIM pose risks, whereas a MF of three times a day and higher MY are protective factors against increases in SCC and SCM occurrence in Brazilian Holstein and Jersey cows., Competing Interests: Declaration of Competing Interest The authors declare no conflict of interest., (Copyright © 2024 Elsevier B.V. All rights reserved.)

In this study, we aimed to improve current udder health genetic evaluations by addressing the limitations of monthly sampled somatic cell score (SCS) for distinguishing cows with robust innate immunity from those susceptible to chronic infections. The objectives were to (1) establish novel somatic cell traits by integrating SCS and the differential somatic cell count (DSCC), which represents the combined proportion of polymorphonuclear leukocytes and lymphocytes in somatic cells and (2) estimate genetic parameters for the new traits, including their daily heritability and genetic correlations with milk production traits and SCS, using a random regression test-day model (RRTDM). We derived 3 traits, termed ML&#95;SCS&#95;DSCC, SCS&#95;4&#95;DSCC&#95;65&#95;binary, and ML&#95;SCS&#95;DSCC&#95;binary, by using milk loss (ML) estimates at corresponding SCS and DSCC levels, thresholds established in previous studies, and a threshold established from milk loss estimates, respectively. Data consisted of test-day records collected during January 2021 through March 2022 from 265 herds in Hokkaido, Japan. From these records, we extracted records between 7 to 305 d in milk (DIM) in the first lactation to fit the RRTDM. The model included the random effect of herd-test-day, the fixed effect of year-month, fixed lactation curves nested with calving age groups, and random regressions with Legendre polynomials of order 3 for additive genetic and permanent environmental effects. The analysis was performed using Gibbs sampling with Gibbsf90+ software. The averages (ranges) of the daily heritability estimates over lactation were 0.086 (0.075-0.095) for SCS, 0.104 (0.073-0.127) for ML&#95;SCS&#95;DSCC, 0.137 (0.014-0.297) for SCS&#95;4&#95;DSCC&#95;65&#95;binary, and 0.138 (0.115-0.185) for ML&#95;SCS&#95;DSCC&#95;binary; the heritability curve for SCS&#95;4&#95;DSCC&#95;65&#95;binary was erratic. Genetic correlations within the trait decreased as the DIM interval widened, especially for those integrating DSCC, indicating that these traits should be analyzed using RRTDM rather than repeatability models. The averages (ranges) of genetic correlations with milk yield over lactation were 0.01 (-0.22 to 0.28) for SCS, -0.05 (-0.40 to 0.13) for ML&#95;SCS&#95;DSCC, -0.08 (-0.17 to 0.09) for SCS&#95;4&#95;DSCC&#95;65&#95;binary, and -0.08 (-0.22 to 0.27) for ML&#95;SCS&#95;DSCC&#95;binary. Compared with SCS, the newly defined traits exhibited slightly stronger negative genetic correlations with milk yield. Especially in late lactation stages, the genetic correlation between ML&#95;SCS&#95;DSCC and milk yield was significantly below zero, with a posterior median of -0.40. Furthermore, the new traits showed positive correlations with SCS, having estimates varying from 0.68 to 0.85 for ML&#95;SCS&#95;DSCC, 0.14 to 0.47 for SCS&#95;4&#95;DSCC&#95;65&#95;binary, and 0.61 to 0.66 for ML&#95;SCS&#95;DSCC&#95;binary, depending on DIM. Considering that ML&#95;SCS&#95;DSCC and ML&#95;SCS&#95;DSCC&#95;binary have relatively high heritability (compared with SCS) and favorable genetic correlations with milk production traits and SCS, their incorporation into breeding programs appears promising. Nevertheless, their genetic relationships with (sub)clinical mastitis require further investigation., (The Authors. Published by Elsevier Inc. on behalf of the American Dairy Science Association®. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).)

Biosecurity plays a crucial role in preventing the introduction of infectious diseases to a herd as well as the spread of diseases within or between animals and herds. In particular, biosecurity measures are crucial for maintaining animal health and reducing the need for the application of antibiotic substances for fighting the rising antibiotic resistance. The object of this cross-sectional study was to investigate the presence of biosecurity measures and their association with milk quality parameters, with a special focus on somatic cell count (SCC) - an indicator for udder health -, in small-scale mountain dairy farms. Therefore, for the very first time, the CLASSYFARM system was considered, which is a computer platform integrated into the Italian national veterinary portal, that processes a significant amount of data from various sources collected in the field or from other information systems (e.g. animal welfare, health status, biosecurity, antimicrobial use, slaughterhouse information). A total of 169 dairy farms were included in the study. Biosecurity measures, based on 15 questions required in the CLASSYFARM welfare assessment protocol, as well as information about husbandry systems, milking systems and pasture practices were gathered and combined with milk yield data, provided by the South Tyrolean dairy association. Farms only scored 44.00 points on average in a scoring system from 0.00 to 100.00 points that was be able to summarize 15 different biosecurity measures in one index. Our results show a clear negative correlation (-0.713) between the biosecurity index and somatic cell score (SCS) indicating that a higher level of biosecurity, which reflects the presence of biosecurity measures within a farm, is associated with lower SCC levels. Furthermore, we found significant correlations between SCS and milk production (-0.629), confirming that udder health is linked to higher milk production. Fat, protein, and the fat-to-protein ratio showed a positive correlation with SCS (0.281, 0.146, 0.106), likely to be caused by a concentration shift effect (dilution effect). Husbandry system, breed, milking system, and pasture practices seem to have an impact as well, but the main factor was the biosecurity score. This study highlights the importance of implementing biosecurity measures for ensuring animal health and thus productivity and quality in milk production, even in small-scale farms, which are characterized by limited structure availability and smaller herds compared to big dairy enterprises in the lowlands., (Copyright © 2024 The Authors. Published by Elsevier B.V. All rights reserved.)

Background: In dairy cattle, mastitis causes high financial losses and impairs animal well-being. Genetic selection is used to breed cows with reduced mastitis susceptibility. Techniques such as milk cell flow cytometry may improve early mastitis diagnosis. In a highly standardized in vivo infection model, 36 half-sib cows were selected for divergent paternal Bos taurus chromosome 18 haplotypes (Q vs. q) and challenged with Escherichia coli for 24 h or Staphylococcus aureus for 96 h, after which the samples were analyzed at 12 h intervals. Vaginal temperature (VT) was recorded every three minutes. The objective of this study was to compare the differential milk cell count (DMCC), milk parameters (fat %, protein %, lactose %, pH) and VT between favorable (Q) and unfavorable (q) haplotype cows using Bayesian models to evaluate their potential as improved early indicators of differential susceptibility to mastitis., Results: After S. aureus challenge, compared to the Q half-sibship cows, the milk of the q cows exhibited higher PMN levels according to the DMCC (24 h, p < 0.001), a higher SCC (24 h, p < 0.01 and 36 h, p < 0.05), large cells (24 h, p < 0.05) and more dead (36 h, p < 0.001) and live cells (24 h, p < 0.01). The protein % was greater in Q milk than in q milk at 0 h (p = 0.025). In the S. aureus group, Q cows had a greater protein % (60 h, p = 0.048) and fat % (84 h, p = 0.022) than q cows. Initially, the greater VT of S. aureus-challenged q cows (0 and 12-24 h, p < 0.05) reversed to a lower VT in q cows than in Q cows (48-60 h, p < 0.05). Additionally, the following findings emphasized the validity of the model: in the S. aureus group all DMCC subpopulations (24 h-96 h, p < 0.001) and in the E. coli group nearly all DMCC subpopulations (12 h-24 h, p < 0.001) were higher in challenged quarters than in unchallenged quarters. The lactose % was lower in the milk samples of E. coli-challenged quarters than in those of S. aureus-challenged quarters (24 h, p < 0.001). Between 12 and 18 h, the VT was greater in cows challenged with E. coli than in those challenged with S. aureus (3-h interval approach, p < 0.001)., Conclusion: This in vivo infection model confirmed specific differences between Q and q cows with respect to the DMCC, milk component analysis results and VT results after S. aureus inoculation but not after E. coli challenge. However, compared with conventional milk cell analysis monitoring, e.g., the global SCC, the DMCC analysis did not provide refined phenotyping of the pathogen response., (© 2024. The Author(s).)

Mastitis is a multifactorial inflammatory disease. The increase in antibiotic resistance of bacteria that cause mastitis means that cattle breeders would prefer to reduce the use of antibiotics. Recently, therapies using mesenchymal stem cells (MSCs) from various sources have gained significant interest in the development of regenerative medicine in humans and animals, due to their extraordinary range of properties and functions. The aim of this study was to analyze the effectiveness of an allogeneic stem cells derived from bone marrow (BMSC) and adipose tissue (ADSC) in treating mastitis in dairy cattle. The research material consisted of milk and blood samples collected from 39 Polish Holstein-Friesian cows, 36 of which were classified as having mastitis, based on cytological evaluation of their milk. The experimental group was divided into subgroups according to the method of MSC administration: intravenous, intramammary, and intravenous + intramammary, and according to the allogeneic stem cells administered: BMSC and ADSC. The research material was collected at several time intervals: before the administration of stem cells, after 24 and 72 h, and after 7 days. Blood samples were collected to assess hematological parameters and the level of pro-inflammatory cytokines, while the milk samples were used for microbiological assessment and to determine the somatic cells count (SCC). The administration of allogeneic MSCs resulted in a reduction in the total number of bacterial cells, Staphylococcus aureus, bacteria from the Enterobacteriaceae group, and a systematic decrease in SCC in milk. The therapeutic effect was achieved via intravenous + intramammary or intramammary administration., (© 2024. The Author(s).)

Background: Bovine mastitis is one of the most widespread diseases affecting cattle, leading to significant losses for the dairy industry. Currently, the so-called gold standard in mastitis diagnosis involves determining the somatic cell count (SCC). Apart from a number of advantages, this method has one serious flaw: It does not identify the etiological factor causing a particular infection, making it impossible to introduce targeted antimicrobial therapy. This can contribute to multidrug-resistance in bacterial species. The diagnostic market lacks a test that has the advantages of SCC and also recognizes the species of pathogen causing the inflammation. Therefore, the aim of our study was to develop a lateral flow immunoassay (LFIA) based on elongation factor Tu for identifying most prevalent Gram-positive cocci responsible for causing mastitis including Streptococcus uberis, Streptococcus agalactiae and Staphylococcus aureus., Results: As a result, we showed that the assay for S. uberis detection demonstrated a specificity of 89.02%, a sensitivity of 43.59%, and an accuracy of 80.3%. In turn, the second variant - assay for Gram-positive cocci reached a specificity of 95.59%, a sensitivity of 43.28%, and an accuracy of 78.33%., Conclusions: Our study shows that EF-Tu is a promising target for LFIA and we have delivered evidence that further evaluation could improve test parameters and fill the gap in the mastitis diagnostics market., (© 2024. The Author(s).)

Objective: Four parameters of a decision tree for Selective Dry Cow Treatment (SDCT), examined in a previous study, were analyzed regarding their efficacy in detecting cows for dry cow treatment (DCT, use of intramammary antimicrobials). This study set out to review wether all parameters (somatic cell count [SCC≥ 200 000 SC/ml 3 months' milk yield recordings prior dry off (DO)], clinical mastitis history during lactation [≥1 CM], culturing [14d prior DO, detection of major pathogens] and California-Mastitis-Test [CMT, > rate 1/+ at DO]) are necessary for accurate decision making, whether there are possible alternatives to replace culturing, and whether a simplified model could replace the decision tree., Material and Methods: Records of 18 Bavarian dairy farms from June 2015 to August 2017 were processed. Data analysis was carried out by means of descriptive statistics, as well as employing a binary cost sensitive classification tree and logit-models. For statistical analyses the outcomes of the full 4-parameter decision tree were taken as ground truth., Results: 848 drying off procedures in 739 dairy cows (C DO ) were included. SCC and CMT selected 88.1%, in combination with CM 95.6% of the cows that received DCT (n=494). Without culturing, 22 (4.4%) with major pathogens (8x Staphylococcus [ S. ] aureus ) infected C DO would have been misclassified as not needing DCT. The average of geometric mean SCC (within 100 d prior DO) for C DO with negative results in culturing was<100 000 SC/ml milk, 100 000-150 000 SC/ml for C DO infected with minor pathogens, and ≥ 150 000 SC/ml for C DO infected with major pathogens (excluding S.aureus ). Using SCC during lactation (at least 1x > 200 000 SC/ml) and positive CMT to select C DO for DCT, contrary to the decision tree, 37 C DO (4.4%) would have been treated "incorrectly without" and 43 C DO (5.1%) "unnecessarily with" DCT. Modifications were identified, such as SCC<131 000 SC/ml within 100 d prior to DO for detecting C DO with no growth or minor pathogens in culturing. The best model for grading C DO for or against DCT (C DO without CM and SCC<200 000 SC/ml [last 3 months prior DO]) had metrics of AUC=0.74, Accuracy=0.778, balanced Accuracy=0.63, Sensitivity=0.92 and Specificity=0.33., Conclusions: Combining the decision tree's parameters SCC, CMT and CM renders suitable selection criteria under the conditions of this study. When omitting culturing, lower thresholds for SCC should be considered for each farm individually to select C DO for DCT. Nonetheless, the most accurate model could not replace the full decision tree., Competing Interests: Es besteht kein Interessenkonflikt., (Thieme. All rights reserved.)

Host response to invasive microbes in the bovine udder has an important role on the animal health and is essential to the dairy industry to ensure production of high-quality milk and reduce the mastitis incidence. To better understand the biology behind these host-microbiome interactions, we investigated the somatic cell proteomes at quarter level for four cows (collected before and after milking) using a shotgun proteomics approach. Simultaneously, we identified the quarter microbiota by amplicon sequencing to detect presence of mastitis pathogens or other commensal taxa. In total, 32 quarter milk samples were analyzed divided in two groups depending on the somatic cell count (SCC). The high SCC group (>100,000 cell/mL) included 10 samples and significant different proteome profiles were detected. Differential abundance analysis uncovers a specific expression pattern in high SCC samples revealing pathways involved in immune responses such as inflammation, activation of the complement system, migration of immune cells, and tight junctions. Interestingly, different proteome profiles were also identified in quarter samples containing one of the two mastitis pathogens, Staphylococcus aureus and Streptococcus uberis, indicating a different response of the host depending on the pathogen. Weighted correlation network analysis identified three modules of co-expressed proteins which were correlated with the SCC in the quarters. These modules contained proteins assigned to different aspects of the immune response, but also amino sugar and nucleotide sugar metabolism, and biosynthesis of amino acids. The results of this study provide deeper insights on how the proteome expression changes at quarter level in naturally infected cows and pinpoint potential interactions and important biological functions during host-microbe interaction., (© 2023. The Author(s).)

Lymphocytes in the colostrum play many important roles during lactation, including protecting newborn piglets against infections. The lymphocytes constantly enter the mammary gland from the mother's bloodstream before and during lactation. However, little is known about the mechanism of transport of maternal lymphocytes across the mammary glands into the milk (lumen). In this study, the maternal lymphocytes were detected in sow colostrum by immunofluorescent staining and fluorescence-activated cell sorting and lymphocytes were observed transmigrating into the breast acinar lumen. Furthermore, immunohistochemical staining revealed that CD3 + T, γδ + T, and IgA + B cells were primarily located at the base area of the mammary gland. Meanwhile, more lactating alveoli and blood capillaries were distributed in this area. Finally, a mammary epithelial cell (EpH4-Ev)/T cell co-culture system was established to explore the mechanism of lymphocyte transmigration across the mammary epithelial cells. The expression of CCL2 and CCL28 in EpH4-Ev cells, which facilitated the transmigration of lymphocytes, significantly increased in the presence of prolactin. Our results provide a better understanding of the concept of lactogenic immunity and pave the way for vaccination strategies for the induction of lactogenic immunity in pregnant swine., (Copyright © 2021 Elsevier B.V. All rights reserved.)

The determination of the somatic cell count of a milk sample is one of the most common methods to monitor udder health of a dairy cow. However, this procedure does not take into account the fact that cells in milk present a great variety of different cell types. The objective of our study was to establish a high-resolution differential cell count (HRDCC) by means of flow cytometry in blood and milk. We were able to detect ten subpopulations among the three main populations of immune cells and to determine their viability. Additionally, blood samples were analyzed for common laboratory biomarkers, i.e. differential blood counts, haptoglobin levels and several metabolic parameters. In this first feasibility study, we used three different vaccines to stimulate the immune system of five healthy cows each. Samples were collected shortly before, in between and after the vaccinations. Using multivariate statistical methods we saw a diagnostic benefit when HRDCCs were included compared to only the standard laboratory parameters. The impacts of all three vaccinations on the immune system were visible in blood HRDCCs as well as in milk HRDCCs. Cluster of Differentiation 8 + (CD8 + ) T cells, B cells and monocyte/macrophage subpopulations were among the most important and statistically relevant parameters for all treatments in both biofluids. Moreover, in one of the treatment groups intermediate monocytes showed a significant increase after both vaccinations. Although the use of HRDCC in blood or milk was shown to be highly relevant for early systemic diagnostic, to confirm these subpopulations further investigations in cows of different breed, lactation stage or health status are required., (© 2021. The Author(s).)

Extracellular vesicles (EVs) have demonstrated unique advantages in serving as nanocarriers for drug delivery, yet the cargo encapsulation efficiency is far from expectation, especially for hydrophilic chemotherapeutic drugs. Besides, the intrinsic heterogeneity of EVs renders it difficult to evaluate drug encapsulation behaviour. Inspired by the active drug loading strategy of liposomal nanomedicines, here we report the development of a method, named "Sonication and Extrusion-assisted Active Loading" (SEAL), for effective and stable drug encapsulation of EVs. Using doxorubicin-loaded milk-derived EVs (Dox-mEVs) as the model system, sonication was applied to temporarily permeabilize the membrane, facilitating the influx of ammonium sulfate solution into the lumen to establish the transmembrane ion gradient essential for active loading. Along with extrusion to downsize large mEVs, homogenize particle size and reshape the nonspherical or multilamellar vesicles, SEAL showed around 10-fold enhancement of drug encapsulation efficiency compared with passive loading. Single-particle analysis by nano-flow cytometry was further employed to reveal the heterogeneous encapsulation behaviour of Dox-mEVs which would otherwise be overlooked by bulk-based approaches. Correlation analysis between doxorubicin auto-fluorescence and the fluorescence of a lipophilic dye DiD suggested that only the lipid-enclosed particles were actively loadable. Meanwhile, immunofluorescence analysis revealed that more than 85% of the casein positive particles was doxorubicin free. These findings further inspired the development of the lipid-probe- and immuno-mediated magnetic isolation techniques to selectively remove the contaminants of non-lipid enclosed particles and casein assemblies, respectively. Finally, the intracellular assessments confirmed the superior performance of SEAL-prepared mEV formulations, and demonstrated the impact of encapsulation heterogeneity on therapeutic outcome. The as-developed cargo-loading approach and nano-flow cytometry-based characterization method will provide an instructive insight in the development of EV-based delivery systems., (© 2021 The Authors. Journal of Extracellular Vesicles published by Wiley Periodicals, LLC on behalf of the International Society for Extracellular Vesicles.)

Compost bedded pack barns (compost) as a new free walk housing system favorably influence udder health due to improved animal welfare and lying comfort. On the other hand, unfavorable effects on udder health are possible, due to the open bedded pack and the associated larger bacterial content in moisture. For in-depth farming system comparisons, the present study aimed to evaluate the specific cell fractions and mastitis pathogens in milk from cows kept in compost and in conventional cubical barns (cubicle). For milk sample collection we used a repeated measurement data structure of 2,198 udder quarters from 537 Holstein cows kept in six herds (3 in compost and 3 in cubicle). Differential cell counting was conducted including lymphocytes, macrophages and polymorphonuclear leukocytes (PMN). Specific mastitis pathogens comprised major and minor pathogens. Mixed models were applied to infer environmental and cow associated effects on cell fractions and on prevalences for pathogen infections, with specific focus on system × lactation stage, system × milk yield and system × somatic cell count effects. The interaction between system and lactation stage showed significant differences (P < 0.01) between the systems. A significantly smaller number of bacteriologically positive quarters and lower prevalences for minor pathogens were detected in compost compared to cubicle. Least squares means for pathogen prevalences indicated a quite constant proportion of bacteriologically negative udder quarters across milk yield levels in compost, but a slight increase with increasing milk yield in cubicle. Cell fraction responses in both systems differed in relation to the overall bacteriological infection status and farming system particularities. In conclusion, different cell fractions and specific mastitis pathogens should be considered as an indicator for udder health in different production systems, taking into account cow associated factors (lactation stage, milk yield).

This research aims to test the hypothesis that neural therapy (NT) on the auricular branch of the vagus nerve (ABVN) in dairy cows diagnosed with subclinical mastitis (SCM) results in a reduction of the somatic cell count. Therapeutic options for SCM are mostly based on use of antibiotics and often lead to unsatisfactory results. An alternative therapy targeting the anti-inflammatory properties of the vagus nerve showing good efficacy, economic viability and without major side effects would be of considerable interest. Auriculotherapy (AT) was performed using three repeated infiltrations of 8.0 mg (0.4 ml) procaine hydrochloride (2%) at the location of the ear tag associated with the auricular acupuncture point (AAP) of the udder. Some 85 clinically healthy cows from nine dairy farms were sampled for evaluation of quarter somatic cell count (QSCC) on four days (d0, d2, d4 and d6). Quarters with a QSCC > 100 000 cells/ml on d0 were included in the analysis. Over the study period, a total of 784 quarters were analysed, 385 control (CON) quarters from 40 cows and 399 treated (TRE) quarters from 45 cows. Results showed that AT of the ABNV resulted in a significant reduction of the QSCC after three treatments. The effect was independent of bacteriological culture results of the quarter milk samples. The bacteriological cure rate, however, was not influenced by AT. To our knowledge this is the first report of AT reducing QSCC in dairy cows with SCM. Before AT can be regarded as an alternative therapeutic approach, further research should focus on possible long-term effects of AT on the reduction of SCC, any bacterial elimination and the neural pathways of AT in dairy cows with SCM.

Bayesian latent class models were used to estimate the test accuracy (sensitivity (Se), specificity (Sp), and predictive values (NPV and PPV)) of cow-level somatic cell counts (SCC) data, quarter-level Petrifilm® on-farm milk culture, and quarter-level standard milk bacteriology for the identification of quarters that should possibly be treated with antimicrobials at dry off in dairy cows. Data of 282 cows from 9 dairy herds in Québec, Canada, with bulk tank SCC < 250,000 cells/mL were used. Estimated median herd-prevalence of infections that should be treated was 16.2 % (95 % credibility interval (CI): 11.0-22.7). Se and Sp estimates for quarter-milk culture using Petrifilm® were 82.2 % (95 %CI: 74.0-89.5) and 62.0 % (95 %CI: 58.6-65.6), respectively. Se and Sp for quarter-milk standard bacteriology were 67.4 % (95 %CI: 55.8-81.2) and 79.6 % (95 %CI: 76.4-83.0), respectively. Se and Sp of different SCC scenarios and thresholds were estimated. For first parity cows, using only the last Dairy Herd Improvement (DHI) test SCC with a threshold of 100,000 cells/mL appeared quite accurate, with Se, Sp, PPV, NPV and reduction of antimicrobial usage of 85.6 % (95 %CI: 69.6-95.6), 86.0 % (95 %CI: 80.0-91.7), 58.0 % (95 %CI: 42.3-74.2), 96.4 % (95 %CI: 91.3-99.0), and 75.3 % (95 %CI: 70.7-79.3), respectively. For cows of ≥ 2nd parity, using only the last DHI test SCC with a threshold of 200,000 cells/mL resulted in Se, Sp, PPV, NPV and reduction of antimicrobial usage of 75.3 % (95 %CI: 55.8-87.3), 84.0 % (95 %CI: 78.8-89.3), 47.2 % (95 %CI: 32.0-63.7), 94.7 % (95 %CI: 89.0-97.6), and of 77.0 % (95 %CI: 73.3-80.3), respectively. Adding quarter-level milk culture using Petrifilm® to cows identified as unhealthy using cow-level SCC data improved the test accuracy (mainly the PPV) and further reduced the use of antimicrobials. For instance, in ≥ 2nd parity cows, using only the last DHI SCC with a threshold of 200,000 cells/mL, adding a subsequent Petrifilm® test increased the reduction from 77.0 % (95 %CI: 73.3-80.3) to 89.5 % (95 %CI: 86.7-91.8). Considering the availability of SCC data, the easiness of using just the last DHI test, and the high NPV that could be achieved, producers may consider using just the last DHI test as a potential tool to identify cows that should be treated with antimicrobials at dry off. It may be used alone or in combination with quarter-level on-farm Petrifilm® milk culture on high SCC cows to further reduce the use of antimicrobials by identifying quarters that need to be treated., (Copyright © 2021 Elsevier B.V. All rights reserved.)

Mastitis is one of the most frequently encountered diseases in dairy cattle, negatively affecting animal welfare and milk production. For this reason, contributions to understanding its genomic architecture are of great interest. Genome-wide association studies (GWAS) have identified multiple loci associated with somatic cell score (SCS) and mastitis in cattle. However, most of the studies have been conducted in different parts of the world on various breeds, and none of the investigations have studied the genetic architecture of mastitis in Romanian dairy cattle breeds up to this point in time. In this study, we report the first GWAS for SCS in dairy cattle breeds from Romania. For GWAS, we used an Axiom Bovine v3 SNP-chip (>63,000 Single Nucleotide Polymorphism -SNPs) and 33,330 records from 690 cows belonging to Romanian Spotted (RS) and Romanian Brown (RB) cattle. The results found one SNP significantly associated with SCS in the RS breed and 40 suggestive SNPs with -log 10 ( p ) from 4 to 4.9 for RS and from 4 to 5.4 in RB. From these, 14 markers were located near 12 known genes ( AKAP8 , CLHC1 , MEGF10 , SATB2 , GATA6 , SPATA6 , COL12A1 , EPS8 , LUZP2 , RAMAC , IL12A and ANKRD55) in RB cattle, 3 markers were close to ZDHHC19 , DAPK1 and MMP7 genes, while one SNP overlapped the HERC3 gene in RS cattle. Four genes ( HERC3 , LUZP2 , AKAP8 and MEGF10 ) associated with SCS in this study were previously reported in different studies. The most significant SNP (rs110749552) associated with SCS was located within the HERC3 gene. In both breeds, the SNPs and position of association signals were distinct among the three parities, denoting that mastitis is controlled by different genes that are dependent according to parity. The current results contribute to an expansion in the body of knowledge regarding the proportion of genetic variability explained by SNPs for SCS in dairy cattle.

Backgrounds: The present study explored the viability of bovine milk macrophages, their intracellular production of reactive oxygen and nitrogen species (RONS), and their phagocytosis of Staphylococcus aureus, as well as the profile of lymphocytes, from healthy udder quarters and udder quarters infected by Corynebacterium bovis. The study included 28 healthy udder quarters from 12 dairy cows and 20 udder quarters infected by C. bovis from 10 dairy cows. The percentages of macrophages and lymphocytes were identified by flow cytometry using monoclonal antibodies. Macrophage viability, RONS production, and S. aureus phagocytosis were evaluated by flow cytometry., Results: Milk samples from quarters infected with C. bovis showed a lower percentage of macrophages but an increased number of milk macrophages per mL and a higher percentage of macrophages that produced intracellular RONS and phagocytosed S. aureus. No effect of C. bovis infection on macrophage viability was found. Udder quarters infected by C. bovis showed a higher percentage of T cells and CD4 + T lymphocytes, but no effect was found on the percentage of CD8 + CD4 - T, CD8 - CD4 - T, or B lymphocytes., Conclusions: Thus, our results corroborate, at least in part, the finding that intramammary infections by C. bovis may offer protection against intramammary infections by major pathogens., (© 2021. The Author(s).)

Variations in the levels of acute phase proteins and lactoferrin in serum and milk for diagnosis of subclinical mastitis in dairy cows are described in this research paper. Milking animals from two organized dairy farms in Kerala, India, were screened by California Mastitis Test (CMT), Electrical Conductivity test (EC) and Somatic Cell Count (SCC) test to identify animals affected with sub clinical mastitis (SCM). The concentrations of acute phase proteins (APP) Haptoglobin (Hp), C- reactive protein (CRP), Albumin, Lactoferrin (Lf) and α- 1 acid glycoprotein (AGP) in milk and Hp, Albumin, Serum Amyloid A (SAA) and CRP in the serum of 40 normal cows and 40 cows affected with sub clinical mastitis were assessed. Solid phase ELISA was employed for assessment of all parameters except the albumin levels, for which spectrophotometry was used. The values of Hp in milk; and SAA, AGP and Lf in serum, were significantly elevated in the group with sub clinical mastitis. Such variations were found to be independent of the specific bacterial organism causing the disease. These results show that significant variations exist in the levels of acute phase proteins Hp, AGP and Lf in milk, and SAA in serum of animals affected with subclinical bovine mastitis that are not affected by specific bacterial etiology.

The current study reports the identification of previously undiscovered single-nucleotide polymorphisms (SNPs) in the bovine AGPAT3 gene and further investigates their associations with milk production traits. Our results demonstrate that the major allele C of the SNP g.12264 C > T is positively correlated with test-day milk yield, protein percentage and 305-day milk yield. Importantly, in silico analysis showed that the C/T transition at this locus gives rise to two new transcription factor binding sites (TFBS), E2F1 and Nkx3-2. Polymorphism g.18658 G > A was the only SNP associated with milk urea nitrogen (MUN) with the G allele related to an increase in milk urea nitrogen as well as fat percentage. The GG genotype of SNP g.28731 A > G was associated with the highest fat and protein percentage and lowest 305-day milk yield and somatic cell score (SCS). The association between AGPAT3 locus and milk production traits could be utilized in marker-assisted selection for the genetic improvement of milk production traits and, probably in conjunction with other traits, for selection to improve fitness of dairy cattle.

This Research Communication describes the relation between somatic cells and microbial content in milk from Jersey cattle. Milk samples were classified in groups: healthy, dirty and mastitic (from Staphylococcus spp., Escherichia coli, Coliforms). The somatic cells in each of those groups were analysed by two methods - flow cytometric and automatic fluorescent cell counting. Those methods were compared. Total somatic cell count (SCC), neutrophil count, and lymphocytes with cluster of differentiation 4 (CD4+cells) were determined. There was a positive relationship between microbes and somatic cells. It was noticed that the neutrophil count was generally increased together with SCC, whilst the CD4+ cell count was higher in healthy milk samples (about 8%) compared to mastitic ones (about 3%). Lower number of CD4+ cells (from 1 to 4%) was determined in samples positive for Staphylococcus spp. but with lower SCC (from 2.7 to 4.0 × 105 cells/ml). Also, the number of CD4+ cells in Staphylococcus spp.-positive samples increased (to 4.8%) together with higher SCC, something that was not observed in the other mastitic samples. Knowledge of those relations could be useful for veterinary medical tests in the initial phase of inflammation.

Dairy goat farming is an important sector of the agricultural industry in Greece, with an annual total milk production exceeding 450 000 l and accounting for over 25% of all goat milk produced in the European Union; this milk is used mainly for cheese production. Despite the importance of goat milk for the agricultural sector in Greece, no systematic countrywide investigations in the bulk-tank milk of goats in Greece have been reported. Objectives were to investigate somatic cell counts (SCC) and total bacterial counts (TBC) in raw bulk-tank milk of goat herds in Greece, study factors influencing SCC and TBC therein and evaluate their possible associations with milk content. Throughout Greece, 119 dairy goat herds were visited for milk sampling for somatic cell counting, microbiological examination and composition measurement. Geometric mean SCC and TBC were 0.838 × 106 cells ml-1 and 581 × 103 cfu ml-1, respectively. Multivariable analyses revealed annual frequency of check-ups of milking system and total milk quantity per goat (among 53 variables) to be significant for increased SCC; no factor emerged (among 58 variables) to be significant for increased TBC. Negative correlation of SCC with total protein was found; mean total protein content in the bulk-tank milk in herds with SCC >0.75 × 106 cells ml-1 was 5.1% lower and in herds with SCC >1.5 × 106 cells ml-1, it was 7.8% lower.

Subclinical mastitis (SCM) in water buffalo is a production disease associated with decreased milk yield and impaired milk quality and safety. Water buffalo is an important livestock species in Bangladesh, but information about the occurrence and aetiology of SCM in this species is scarce. A cross-sectional study was conducted as part of the Udder Health Bangladesh Programme to (i) determine the occurrence of SCM and bulk milk somatic cell count (SCC) in water buffalo in Bangladesh, (ii) identify pathogens causing SCM and (iii) evaluate penicillin resistance in isolated staphylococci strains. Sixteen buffalo farms in the Bagerhat and Noakhali regions of Bangladesh were selected for study and a bulk milk sample was collected from each farm. In addition, 299 udder quarter milk samples were collected from 76 animals. The bulk milk samples were assessed by direct SCC and the quarter milk samples by California mastitis test (CMT). The occurrence of SCM calculated at quarter and animal level was 42.5 and 81.6%, respectively. Milk samples from 108 CMT-positive quarters in 48 animals and 38 randomly selected CMT-negative quarters in 24 animals were investigated using bacteriological culture. Estimated mean bulk milk SCC was 195 000 cells/ml milk (range 47 000- 587 000 cells/ml milk). On culture, estimated quarter-level intramammary infection (IMI) was 40.4%. The identity of isolated bacteria was confirmed by MALDI-TOF mass spectrometry. Non-aureus staphylococci (NAS) were the most common pathogens (24.7%) and, among 36 NAS tested, 36.1% were resistant to penicillin. Thus there was high occurrence of SCM on the study farms, with relatively high penicillin resistance in NAS. Further studies are needed to identify underlying risk factors and develop an udder health control strategy for water buffalo in Bangladesh.

This research communication describes the application of a fluorescent automatic cell counter Lactoscan SCC for simultaneous determination of somatic cell count and neutrophils in bovine milk. The obtained results were compared with results obtained by a flow cytometer and a light microscope. The Pearson correlations between the methods were calculated. A comparison between the main characteristics of the three kinds of analysis was made - the assay duration and the intra-assay precision. A relation between the SCC and neutrophil cells was observed in 55 milk samples. The obtained results confirm that the simultaneous determination of SCC and neutrophil analysis are necessary and support the early diagnosis of mastitis, the timely treatment of the animal and the avoidance of major economic losses.

Bovine mammary function at molecular level is often studied using mammary tissue or primary bovine mammary epithelial cells (pbMECs). However, bulk tissue and primary cells are heterogeneous with respect to cell populations, adding further transcriptional variation in addition to genetic background. Thus, understanding of the variation in gene expression profiles of cell populations and their effect on function are limited. To investigate the mononuclear cell composition in bovine milk, we analyzed a single-cell suspension from a milk sample. Additionally, we harvested cultured pbMECs to characterize gene expression in a homogeneous cell population. Using the Drop-seq technology, we generated single-cell RNA datasets of somatic milk cells and pbMECs. The final datasets after quality control filtering contained 7,119 and 10,549 cells, respectively. The pbMECs formed 14 indefinite clusters displaying intrapopulation heterogeneity, whereas the milk cells formed 14 more distinct clusters. Our datasets constitute a molecular cell atlas that provides a basis for future studies of milk cell composition and gene expression, and could serve as reference datasets for milk cell analysis., (© 2021. The Author(s).)

Evidence suggests that dietary depletion of bovine milk exosomes and their cargos causes a loss of circulating microRNAs and a series of health problems. The aim of the current study was to determine whether bovine milk exosomes affect purine nucleotide metabolism and energy metabolism in oxidatively stressed intestinal crypt epithelial cells (IEC-6). Cells were pretreated with exosomes, followed by H 2 O 2 to induce oxidative stress. Reactive oxidative species (ROS) levels, purine nucleotides, purine metabolic key enzyme activities, cell energy status, and AMPK protein expression were analysed. Exosome pretreatment reduced ROS level and the activities of adenosine deaminase and xanthine oxidase induced by H 2 O 2 in cells. Total adenine nucleotides and energy charge were increased with exosome pretreatment, while the AMPK phosphorylation level was downregulated. The results indicated that bovine milk exosomes could attenuate purine nucleotide catabolism and improve energy status in oxidatively stressed IEC-6 cells and exerted protective effects against oxidative stress., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

Mastitis, in particular in its subclinical form, which may spread unnoticeable within a herd, continues to be a major challenge in the dairy industry. Somatic cell count (SCC) is a broadly used proxy for subclinical mastitis. The recently introduced Differential SCC (DSCC) representing the combined proportion of polymorphonuclear neutrophils and lymphocytes as a percentage of total SCC, can be used in combination with SCC to categorise cows into four different udder health groups (UHG) depending on actual test day results: UHG A: healthy/normal, ≤200,000 cells/mL and DSCC ≤65 %; B: suspicious, ≤200,000 cells/mL and DSCC >65 %; C: (subclinical) mastitis, >200,000 cells/mL and DSCC >65 %; D: chronic/persistent mastitis, >200,000 cells/mL and DSCC ≤65 %. The objective of our study was to investigate to what extent the UHG aid in determining different statuses of cows: I) leaving herd before next test day, II) having >200,000 cells/mL at the next test day, and III) having ≤200,000 cells/mL at the next 3 test days. Multivariable logistic regression analysis was used to evaluate these statuses based on routinely generated dairy herd improvement (DHI) data from Austria, China, Estonia, Germany, and Spain. Cows in groups C (odds ratio (OR): 2.13, 95 % confidence interval (CI): 1.95-2.34) and, particularly, D (OR: 3.91, 95 % CI: 3.31-4.62) were significantly more likely to leave herds compared to cows in group A. Late-lactating cows indicated the highest likelihood (OR: 16.03, 95 % CI: 14.44-17.81) to leave herds in our analysis. Interestingly, we found that cows in UHG B had significantly higher odds (OR: 2.77, 95 % CI: 2.58-2.98) to have >200,000 cells/mL at the next test day compared to cows in group A. As anticipated, cows in UHG B (OR: 0.40, 95 % CI: 0.38-0.42), C (OR: 0.08, 95 % CI: 0.07-0.09), and D (OR: 0.16, 95 % CI: 0.14-0.19) each were significantly less likely to have ≤200,000 cells/mL at the next 3 test days compared to cows in group A. Above described results are an example from Germany, but the same trends could be seen across all countries considered in our study. In conclusion, our findings illustrate that the UHG concept reveals additional valuable information about udder health and culling based a single test day over working with SCC only. Actual decisions in day-to-day farm management that could be taken were not investigated here and need to be further explored., (Copyright © 2021 FOSS Analytical A/S. Published by Elsevier B.V. All rights reserved.)

The concept that extracellular vesicles (EVs) from the diet can be absorbed by the intestinal tract of the consuming organism, be bioavailable in various organs, and in-turn exert phenotypic changes is highly debatable. Here, we isolate EVs from both raw and commercial bovine milk and characterize them by electron microscopy, nanoparticle tracking analysis, western blotting, quantitative proteomics and small RNA sequencing analysis. Orally administered bovine milk-derived EVs survive the harsh degrading conditions of the gut, in mice, and is subsequently detected in multiple organs. Milk-derived EVs orally administered to mice implanted with colorectal and breast cancer cells reduce the primary tumor burden. Intriguingly, despite the reduction in primary tumor growth, milk-derived EVs accelerate metastasis in breast and pancreatic cancer mouse models. Proteomic and biochemical analysis reveal the induction of senescence and epithelial-to-mesenchymal transition in cancer cells upon treatment with milk-derived EVs. Timing of EV administration is critical as oral administration after resection of the primary tumor reverses the pro-metastatic effects of milk-derived EVs in breast cancer models. Taken together, our study provides context-based and opposing roles of milk-derived EVs as metastasis inducers and suppressors.

In this research communication we address the hypothesis that a single intramammary infusion of casein hydrolyzate (CH) would have a similar effect to three intramammary infusions of CH for drying-off quarters with chronic mastitis (CM) during lactation. Sixty cows with CM were selected and randomly distributed into two treatment groups: (a) three intramammary CH infusions (100 mg, 50 ml per infusion, with 24-h intervals) or (b) single intramammary CH infusion (300 mg, 50 ml). Milk samples from the treated and untreated quarters were collected for microbiological culture and somatic cell count (SCC) before and after CH infusions. Milk yield was recorded and a manual pressure index measurement was used to evaluate cessation of lactation. Of the 60 quarters selected, 43 (71.67%) had positive microbiological culture. The quarters treated with three intramammary CH infusions had higher udder pressure index than those treated with single CH infusion. However, the average milk yield and composite SCC of three functional quarters were not different among treatments. Therefore, a single infusion of CH has the potential to be used as an alternative method for drying-off mammary quarters with CM during lactation.

This research paper focuses on time-lagged heat stress (HS) effects from an across-generation perspective. Temperature × humidity indexes (THI) from the last 8 weeks of pregnancy were associated with subsequent female offspring performances. The offspring dataset considered 172 905 Holstein dairy cows from calving years 2002-2013 from 1,968 herds, located in the German federal state of Hesse. Production traits included milk yield (MKG), protein percentage (PRO%), fat percentage (FAT%), somatic cell score (SCS) and milk urea nitrogen (MUN) from the first official test-day in first lactation. Female fertility traits were the non-return-rate after 56 d (NRR56) in heifers and the interval from calving to first insemination (ICFI) in first parity cows. Longevity traits were the length of productive life (LPL), lifetime productivity in milk yield (LTP-MKG) and milk yield per day of life (MKG-DL). The association analyzes for 10 traits combined with meteorological data from 8 single weeks before calving implied in total 80 different runs. THI ≥50 from all single 8 weeks before calving had unfavorably significant effects on FAT%, ICFI and LPL. Heat stress in terms of THI ≥60 from the last 3 weeks before calving impaired MKG. NRR56 decreased with increasing THI, as observed for all 6 weeks before calving. LTP-MKG and MKG-DL decreased due to high THI in the last 4 weeks before calving. Heat stress (THI ≥60) during late pregnancy had no significantly unfavorable impact on PRO% and MUN. Interestingly, SCS in offspring declined with increasing THI during late pregnancy. In conclusion, for most of the primary and functional traits, unfavorable impact of HS from the dry period on time-lagged performances in offspring was identified, even on longevity. From a practical perspective, our data suggest to provide HS abatement to late gestation dams to avoid long-term adverse effects on the offspring.

We investigated the effects of pathogens associated with subclinical intramammary infections on yield, composition and quality indicators of goat milk. By means of a longitudinal study, individual half udder milk samples (n = 132) were collected at different lactation periods and assessed for milk yield and physicochemical composition, somatic cell count (SCC), total bacteria count (TBC) and microbiological culture. Staphylococci species accounted for the great majority of the isolates (96.1%). Intramammary infections significantly reduced fat and total solids in goat milk and increased both SCC and TBC. However, these indicators were significantly higher in udder halves affected by S. aureus compared with other staphylococci species.

Bovine mastitis, an inflammatory disease of the mammary gland, is classified as subclinical or clinical. Circulating neutrophils are recruited to the udder to combat infection. We compared the transcriptomic profiles in circulating leukocytes between healthy cows and those with naturally occurring subclinical or clinical mastitis. Holstein Friesian dairy cows from six farms in EU countries were recruited. Based on milk somatic cell count and clinical records, cows were classified as healthy (n = 147), subclinically (n = 45) or clinically mastitic (n = 22). Circulating leukocyte RNA was sequenced with Illumina NextSeq single end reads (30 M). Differentially expressed genes (DEGs) between the groups were identified using CLC Genomics Workbench V21, followed by GO enrichment analysis. Both subclinical and clinical mastitis caused significant changes in the leukocyte transcriptome, with more intensive changes attributed to clinical mastitis. We detected 769 DEGs between clinical and healthy groups, 258 DEGs between subclinical and healthy groups and 193 DEGs between clinical and subclinical groups. Most DEGs were associated with cell killing and immune processes. Many upregulated DEGs in clinical mastitis encoded antimicrobial peptides (AZU1, BCL3, CAMP, CATHL1, CATHL2, CATHL4,CATHL5, CATHL6, CCL1, CXCL2, CXCL13, DEFB1, DEFB10, DEFB4A, DEFB7, LCN2, PGLYRP1, PRTN3, PTX3, S100A8, S100A9, S100A12, SLC11A1, TF and LTF) which were not upregulated in subclinical mastitis. The use of transcriptomic profiles has identified a much greater up-regulation of genes encoding antimicrobial peptides in circulating leukocytes of cows with naturally occurring clinical compared with subclinical mastitis. These could play a key role in combatting disease organisms.

Extracellular vesicles (EVs) provide a novel intercellular communication mechanism to transfer biologically important molecules to target cells. Although several pieces of evidence have shown that EVs have potential to respond to bacterial infections, our knowledge about the role of circular RNA (circRNA), an important cargo of EV, behind this process remains poor. In particular, the mechanism by which circRNAs are packaged into EVs remains elusive during bacterial infection. In the present study, EVs from bovine milk samples with or without Staphylococcus aureus ( S. aureus ) infection were isolated. The presence of circRNAs in milk-derived EVs (MEVs) was validated for the first time by PCR amplification with convergent and divergent primers and the RNase R resistance test. Through high-throughput sequencing, the expression profile of circRNAs in EVs was found to be changed during S. aureus infection. Moreover, we demonstrated that circRNAs were selectively packaged into EVs. Finally, bioinformatic analyses predicted the involvement of differentially expressed circRNAs in immune functions. In summary, our findings offer an insight into the packaging mechanism of EV circRNAs and underscore the potential by which host used the EV circRNAs in response to pathogenic bacterial infections.

This study aimed to evaluate the use of thermal imaging obtained by infrared thermography (IRT) to detect cases of subclinical mastitis in dairy cows under commercial conditions of compost barn systems in a region of Brazil with a semiarid climate. Twenty-eight crossbred cows were evaluated twice a day for one week using IRT. Three thermal images were obtained for each cow, referring to the anatomical regions of the right and left fore udder and rear udder. A computer program was used to analyze the images and obtain the right fore udder temperature (RFUT, °C), left fore udder temperature (LFUT, °C), rear udder temperature (RUT, °C), and average udder temperature (AUT, °C). In addition, samples of milk from each quarter of the udder were collected for somatic cell count (SCC) to correlate the diseases observed on the thermal image with any infection in the udder region. The results obtained using IRT were subjected to regression and correlation analyses. It was observed that LFUT, RAQT, RUT, and AUT were adjusted in quadratic polynomial models with good prediction of SCC (i.e., infection) with R 2  = 0.92, 0.97, 0.86, and 0.94, respectively. The region of the anterior quarters of the udder was the most promising for imaging, stronger correlations were obtained between LFUT and RFUT with SCC (r = 0.87 and 0.88, respectively). The IRT is a practical technology capable of detecting cases of mastitis in dairy cows with good precision, especially with thermal images from the anatomical region of the front quarters of the udder. However, more detailed studies are needed to make thermal imaging processing a more useful method for routine activities on farms in compost barn systems., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

Milk small extracellular vesicles (sEV) contain proteins that provide potential information of host physiology and immunology. Bovine leukemia virus (BLV) is an oncogenic virus that causes progressive B-cell lymphosarcoma in cattle. In this study, we aimed to explore the proteomic profile of milk sEV from BLV-infected cattle compared with those from uninfected cattle. Milk sEV were isolated from three BLV-infected and three uninfected cattle. Proteomic analysis was performed by using a comprehensive nanoLC-MS/MS method. Furthermore, gene ontology (GO) annotation and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways were used to evaluate the candidates for uniquely or differentially expressed proteins in milk sEV from BLV-infected cattle. Proteomic analysis revealed a total of 1330 common proteins in milk sEV among BLV-infected cattle, whereas 118 proteins were uniquely expressed compared with those from uninfected cattle. Twenty-six proteins in milk sEV were differentially expressed proteins more than two-fold significant difference (p < 0.05) in BLV-infected cattle. GO and KEGG analyses indicated that the candidates for uniquely or differentially expressed proteins in milk sEV had been involved in diverse biological activities including metabolic processes, cellular processes, respond to stimulus, binding, catalytic activities, cancer pathways, focal adhesion, and so on. Taken together, the present findings provided a novel insight into the proteomes of milk sEV from BLV-infected cattle.

This research communication presents an automatic method for the counting of somatic cells in buffalo milk, which includes the application of a fuzzy clustering method and image processing techniques (somatic cell count with fuzzy clustering and image processing|, SCCFCI). Somatic cell count (SCC) in milk is the main biomarker for assessing milk quality and it is traditionally performed by exhaustive methods consisting of the visual observation of cells in milk smears through a microscope, which generates uncertainties associated with human interpretation. Unlike other similar works, the proposed method applies the Fuzzy C-Means (FCM) method as a preprocessing step in order to separate the images (objects) of the cells into clusters according to the color intensity. This contributes signficantly to the performance of the subsequent processing steps (thresholding, segmentation and recognition/identification). Two methods of thresholding were evaluated and the Watershed Transform was used for the identification and separation of nearby cells. A detailed statistical analysis of the results showed that the SCCFCI method is able to provide results which are consistent with those obtained by conventional counting. This method therefore represents a viable alternative for quality control in buffalo milk production.

The objective of this study was to investigate associations of freestall design and cleanliness with cow lying behavior, hygiene, lameness, and risk of new high somatic cell count (SCC). Cows from 18 commercial freestall dairy herds (22 ± 15 cows/farm; mean ± SD) in Ontario, Canada, were enrolled in a longitudinal study. Four hundred focal cows that were <120 d in milk, had no mastitis treatment in the last 3 mo, and had an SCC <100,000 cells/mL at their most recent milk test were selected for the study. Data on SCC were collected through Dairy Herd Improvement Association milk testing (at ~5-wk intervals). Each farm was visited 5 ± 3 d (mean ± SD) after each milk test until 3 tests were completed (~105 d), for a total of 3 observation periods per cow. Elevated SCC was used as an indicator of subclinical mastitis. An incident of new high SCC was defined as a cow having SCC >200,000 cells/mL at the end of an observation period, when SCC was <100,000 cells/mL at the beginning of that period. Lying behavior was recorded for 6 d after each milk sampling, using electronic data loggers. Cows were scored during each period for lameness (5-point scale, with scores ≥3 = lame), body condition score (BCS; 5-point scale; 1 = thin to 5 = fat), and hygiene (4-point scale). Stall cleanliness was assessed during each period with a 1.20 × 1.65-m metal grid, containing 88 squares. The grid was centered between stall partitions of every tenth stall on each farm, and the squares containing visible urine or fecal matter (or both) were counted. Cow lying time averaged 10.9 ± 1.9 h/d. On average, cows with low BCS (≤2.5) spent 37 ± 16.6 min/d less time lying down than high-BCS cows (≥4.0). On average, cows tended to spend 36 ± 18.3 min/d more time lying down in deep-bedded versus mattress-based stalls. Mean proportion of soiled squares per stall was 20.1 ± 0.50%. Across farms, cow lying time decreased as the proportion of soiled squares per stall increased. A difference in daily lying time of ~80 more min/d was modeled for cows housed in barns with the cleanest stalls compared with those with the dirtiest stalls. Higher neck rail height [for every 1 SD (10 cm) increase] increased the odds (odds ratio = 1.5) of cows having a dirty upper leg-flank and udder. The odds of a cow having a dirty upper leg-flank, udder, and lower legs were 1.5, 2.0, and 1.9 times greater, respectively, for cows housed with dirtier stalls. Also, cows housed on farms with dirtier stalls had 1.3 times greater odds of being lame at the time of observation. Over the study period, 50 new high-SCC cases were detected, resulting in an incidence rate of 0.45 cases of new high SCC per cow-year at risk. No measured factors were detected to be associated with risk of a new high SCC. Overall, our results confirm that cows lie down longer in cleaner and more comfortable environments. Further, these results highlight the need for improved stall cleanliness to optimize lying time and potentially reduce lameness., (Copyright © 2021 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

Our objective was to evaluate the efficacy of intramammary administration, at drying-off, of a Panax ginseng extract (PGe) combined with cephalexin (Ceph) on the post-calving bacteriological cure rate of pre-existing intramammary infections (IMI) and on the occurrence of new IMI during the dry period. In addition, milk yield and somatic cell count (SCC) in the post-treatment lactation were evaluated. One hundred and eight late-lactation cows were randomly divided into two experimental groups and were treated at drying-off with Ceph alone or PGe combined with Ceph.Cure rates for IMI present at drying-off were similar for both treatments (OR = 0.95, 95% CI = 0.33-2.74). Cure rates for Staphylococcus aureus were lower (OR = 15.4, 95% CI = 1.66-142.52) in quarters treated with PGe + Ceph than in those treated with Ceph alone. Intramammary infusion of PGe + Ceph at drying-off had no effect on preventing new dry period IMI (OR = 0.75, 95% CI = 0.38-1.51), compared with infusion of Ceph alone. Milk production and SCC in the ensuing lactation were not affected by PGe + Ceph treatment. In conclusion, addition of PGe to dry cow therapy did not show any advantage over the use of dry cow therapy alone.