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Clinical mastitis (CM) is a prevalent and severe inflammatory disease in dairy cows affecting the mammary glands. Fatty acid (FA) metabolism and associated enzymes are crucial for many physiological and pathological processes in dairy cows. However, the relationships among FA metabolism, FA-associated enzymes, and CM, as well as the mechanisms underlying their interactions, in dairy cows are not fully understood. The aim of this study was to characterize biological process (BP) terms, pathways, and differentially expressed proteins (DEPs) related to FA metabolism from our previous data-independent acquisition proteomic study. Six BPs involving 14 downregulated and 20 upregulated DEPs, and four pathways involving 10 downregulated and 11 upregulated DEPs related to FA synthesis and metabolism were systematically identified. Associated analysis suggested that 12 candidate DEPs obtained from BPs and pathways, especially acyl-CoA thioesterase 7 (ACOT7), regulate long-chain FA (LCFA) elongation and the biosynthesis of unsaturated FAs. Immunohistochemical and immunofluorescence staining results showed that ACOT7 was present mainly in the cytoplasm of mammary epithelial cells. The qRT-PCR and Western blotting results showed that ACOT7 mRNA and protein levels in the mammary glands of the CM group were significantly upregulated compared to those in the healthy group. This evidence indicates that ACOT7 is positively correlated with CM onset and progression in Holstein cows. These findings offer novel insights into the role of FA metabolism and related enzymes in CM and offer potential targets for the development of therapeutic strategies and biomarkers for the prevention and treatment of CM in dairy cows. [ABSTRACT FROM AUTHOR]

Background: In China's expanding dairy industry, a lack of oversight regarding antimicrobial use and increasing antimicrobial resistance are evident. Selective treatments of dairy cows for clinical mastitis or dry cow therapy are proposed to promote judicious antimicrobial use without adversely impacting cattle health. These approaches have been successfully implemented on farms in other countries. Methods: On 28 October 2023, a 2-day in-person seminar was held in Beijing, China, on selective antimicrobial treatments of dairy cows for clinical mastitis or dry cow therapy on large Chinese dairy farms. Concurrently, a qualitative study involving 15 technical managers from the 13 largest Chinese dairy groups used focus group discussions and questionnaires to explore perspectives on selective treatments of dairy cows for clinical mastitis or dry cow therapy. The main outcomes assessed were opinions and concerns regarding implementing selective antimicrobial treatments. Results: Although there was diversity of cognition on AMR and selective treatments, the technical managers were generally positive regarding adoption of selective treatments. However, they expressed a need for more evidence and tools, including anticipated economic impacts, effects of delaying treatment until diagnosis, accurate interpretation of milk recording data, safe use of internal teat sealants, and spread of pathogens. Participants stressed the need for awareness, staff training, farm management, and China-specific standards, suggesting large-scale trials to assess efficacy of selective treatments. Conclusion: The findings revealed key challenges and barriers currently impeding selective AMU practices. These insights could inform efforts to promote judicious AMU on farms through targeted treatment regimens, reducing mounting selective pressure driving resistance. [ABSTRACT FROM AUTHOR]

The serious drawback underlying the biological annotation of whole-genome sequence data is the p >> n problem, which means that the number of polymorphic variants (p) is much larger than the number of available phenotypic records (n). We propose a way to circumvent the problem by combining a LASSO logistic regression with deep learning to classify cows as susceptible or resistant to mastitis, based on single nucleotide polymorphism (SNP) genotypes. Among several architectures, the one with 204,642 SNPs was selected as the best. This architecture was composed of two layers with, respectively, 7 and 46 units per layer implementing respective drop-out rates of 0.210 and 0.358. The classification of the test data resulted in AUC = 0.750, accuracy = 0.650, sensitivity = 0.600, and specificity = 0.700. Significant SNPs were selected based on the SHapley Additive exPlanation (SHAP). As a final result, one GO term related to the biological process and thirteen GO terms related to molecular function were significantly enriched in the gene set that corresponded to the significant SNPs. Our findings revealed that the optimal approach can correctly predict susceptibility or resistance status for approximately 65% of cows. Genes marked by the most significant SNPs are related to the immune response and protein synthesis. [ABSTRACT FROM AUTHOR]

Simple Summary: The enhanced selection index proposed in the study incorporates new health disorder traits related to udder, claw, and reproduction in addition to the current selection traits. Genetic parameters and breeding values for the traits were evaluated using linear animal models and single-step genomic best linear unbiased prediction (ssGBLUP) methods. The economic weights (EWs) for the new health disorders were calculated using the enhanced bioeconomic model implemented in the computer programme EWDC (i.e., economic weights for dairy cattle), which is versatile and can be applied for various breeds, farm management, and conditions. The use of the enhanced selection index mostly allowed favourable selection progress in terms of the new and current breeding objective traits and could be acceptable for local breeders. The aim of this study was to construct an enhanced selection index using the genomic and economic parameters of new health disorders and current production and functional traits. Genomic evaluation for the incidence of clinical mastitis (CM), three claw disease traits, retained placenta (RET), metritis (MET), and cystic ovaries (CYS) was performed using linear animal models based on producer-recorded data. Good correlations among the health disorders were found, and their heritability estimates did not exceed 7%. Economic weights (EWs) for the health disorders were EUR −132.10 for CM, EUR −128.87 for overall claw diseases, EUR −52.10 for RET, EUR −80.48 for MET, and EUR −16.16 for CYS. These EWs indicate changes in the present value of the annual profit per cow when increasing the incidence of the traits by one case per cow year. Selection using the enhanced index resulted in favourable responses for most of the new health disorders (e.g., −0.001 and −0.006 cases of RET and MET per cow year, respectively), and also in the current breeding objective traits (+49 kg of milk, −0.02% of calf losses). An index contribution of 7% for the new health disorders was assessed as acceptable for the breeders. [ABSTRACT FROM AUTHOR]

Simple Summary: The study focused on improving the detection of clinical bovine mastitis, the inflammation of the udder in cows as a response to intramammary infection, which can be identified by the presence of clots in the milk. Currently, automated milking systems do not detect this important disease very accurately. To address this, we developed a clots detection program using a neural network. This neural network was trained to recognize clots in milk samples from dairy cows by using a large number of pictures of milk filter socks, some with and some without clots. These pictures were divided into different sets for training, validating, and testing the program, respectively. The settings of the neural network were optimized using a genetic algorithm. The program's interpretations were explained using a method called integrated gradients. The program was found to be 100% accurate in identifying clots in the test pictures. This suggests that the method could be very useful for automatically checking for clinical mastitis on dairy farms, although further field validation through integration into the existing systems is needed. Automated milking systems (AMSs) already incorporate a variety of milk monitoring and sensing equipment, but the sensitivity, specificity, and positive predictive value of clinical mastitis (CM) detection remain low. A typical symptom of CM is the presence of clots in the milk during fore-stripping. The objective of this study was the development and evaluation of a deep learning model with image recognition capabilities, specifically a convolutional neural network (NN), capable of detecting such clots on pictures of the milk filter socks of the milking system, after the phase in which the first streams of milk have been discarded. In total, 696 pictures were taken with clots and 586 pictures without. These were randomly divided into 60/20/20 training, validation, and testing datasets, respectively, for the training and validation of the NN. A convolutional NN with residual connections was trained, and the hyperparameters were optimized based on the validation dataset using a genetic algorithm. The integrated gradients were calculated to explain the interpretation of the NN. The accuracy of the NN on the testing dataset was 100%. The integrated gradients showed that the NN identified the clots. Further field validation through integration into AMS is necessary, but the proposed deep learning method is very promising for the inline detection of CM on AMS farms. [ABSTRACT FROM AUTHOR]

The aim of this cross-sectional study was to investigate associated factors of the severity of clinical mastitis (CM). Milk samples of 249 cases of CM were microbiologically examined, of which 27.2% were mild, 38.5% moderate, and 34.3% severe mastitis. The samples were incubated aerobically and anaerobically to investigate the role of aerobic and anaerobic microorganisms. In addition, the pathogen shedding was quantitatively examined, and animal individual data, outside temperature and relative humidity, were collected to determine associated factors for the severity of CM. The pathogen isolated the most was Escherichia coli (35.2%), followed by Streptococcus spp. (16.4%). Non-aureus staphylococci (NaS) (15.4%) and other pathogens (e.g., Staphylococcus aureus, coryneforms) (15.4%) were the pathogens that were isolated the most for mild mastitis. Moderate mastitis was mostly caused by E. coli (38%). E. coli was also the most common pathogen in severe mastitis (50.6%), followed by Streptococcus spp. (16.4%), and Klebsiella spp. (10.3%). Obligate anaerobes (Clostridium spp.) were isolated in one case (0.4%) of moderate mastitis. The mortality rate (deceased or culled due to the mastitis in the following two weeks) was 34.5% for severe mastitis, 21.7% for moderate mastitis, and 4.4% for mild mastitis. The overall mortality rate of CM was 21.1%. The pathogen shedding (back logarithmized) was highest for severe mastitis (55,000 cfu/mL) and E. coli (91,200 cfu/mL). High pathogen shedding, low previous somatic cell count (SCC) before mastitis, high outside temperature, and high humidity were associated with severe courses of mastitis. [ABSTRACT FROM AUTHOR]

Mastitis is one of the main diseases that affects dairy cattle. It is characterized by a multifactorial disease with multiple etiologies including microorganisms such as bacteria, viruses, fungi, yeasts, and algae. Mastitis caused by Mycoplasma spp. results in a contagious nature of infection and has been studied much in Brazil. The objectives of this research were (1) to investigate in clinical mastitis milk samples, by conventional PCR, the presence of the following mycoplasma species: Mycoplasma (M.) bovis, M. bovirhinis, M. bovigenitalium, M. californicum, and M. alkalescens and (2) compare the PCR Mollicutes detection rates using previous concentration and homogenization steps of the milk samples. Of the 993 clinical mastitis milk samples analyzed, 13.7% (136/993) and 8.0% (80/993) were positive for the Mollicutes class after concentration and homogenization, respectively, and all of them were PCR-negative for the five species investigated. Of the 70 expansion milk tanks analyzed, all of them were PCR-negative for the Mollicutes class. The importance of the previous concentration of milk in the routine of molecular analysis was evidenced when compared to the results obtained only with homogenization. [ABSTRACT FROM AUTHOR]

Simple Summary: Clinical mastitis (CM) is one of the most prevalent diseases in cows. Anemoside B4, the main active ingredient in P. chinensis, has been shown to reduce the concentration of pro-inflammatory cytokines and is effective in cows with CM. However, the changes in the metabolic profile of cows with CM after treatment with anemoside B4 remain unclear. This study aimed to investigate how anemoside B4 affects the plasma metabolic profile in cows with CM and to provide a theoretical basis for its effects in veterinary practice. Anemoside B4 has a good curative effect on cows with CM; however, its impact on their metabolic profiles is unclear. Based on similar somatic cell counts and clinical symptoms, nine healthy dairy cows and nine cows with CM were selected, respectively. Blood samples were collected from cows with mastitis on the day of diagnosis. Cows with mastitis were injected with anemoside B4 (0.05 mL/kg, once daily) for three consecutive days, and healthy cows were injected with the same volume of normal saline. Subsequently, blood samples were collected. The plasma metabolic profiles were analyzed using untargeted mass spectrometry, and the concentrations of interleukin (IL)-1β, IL-6, and tumor necrosis factor-α (TNF-α) in serum were evaluated via ELISA. The cows with CM showed increased concentrations of IL-1β, IL-6, and TNF-α (p < 0.05). After treatment with anemoside B4, the concentrations of IL-1β, IL-6, and TNF-α were significantly decreased (p < 0.01). Untargeted metabolomics analysis showed that choline, glycocholic acid, PC (18:0/18:1), 20-HETE, PGF3α, and oleic acid were upregulated in cows with CM. After treatment with anemoside B4, the concentrations of PC (16:0/16:0), PC (18:0/18:1), linoleic acid, eicosapentaenoic acid, phosphorylcholine, and glycerophosphocholine were downregulated, while the LysoPC (14:0), LysoPC (18:0), LysoPC (18:1), and cis-9-palmitoleic acid were upregulated. This study indicated that anemoside B4 alleviated the inflammatory response in cows with CM mainly by regulating lipid metabolism. [ABSTRACT FROM AUTHOR]

Compost-bedded pack barns (CB) are receiving increasing attention as a housing system that can potentially improve the welfare of dairy cows. This study characterized the frequency and profile of pathogens isolated from clinical (CM) and subclinical (SCM) mastitis in dairy cows housed in CB. It evaluated the association between mastitis occurrence and bedding characteristics in CB systems. Over six months, seven dairy herds were visited monthly for milk and bedding sample collections. Milk samples from mastitis cases were submitted to microbiological identification by matrix-assisted laser desorption ionization-time of flight (MALDI-TOF MS). Bedding samples were submitted to physical-chemical (pH, organic matter, moisture, and carbon to nitrogen ratio) and microbiological counting (total bacterial counts, coliforms, streptococci, and staphylococci) analyses. Regression analysis was used to determine the association between mastitis occurrence and CB characteristics. Our results showed that Escherichia coli and environmental streptococci were the most frequently isolated pathogens from CM cases, while Staphylococcus chromogenes and contagious pathogens (Staphylococcus aureus and Streptococcus agalactiae) were the most commonly isolated from SCM cases. Bedding moisture content was positively associated with the incidence of CM. The bedding carbon to nitrogen ratio was negatively associated with the incidence of SCM, and the bedding total bacteria counts tended to be associated with the incidence of SCM. Bedding counts of coliforms positively associated with the prevalence of SCM. Our results can support decision-makers in the dairy industry seeking strategies for bedding management and mastitis control. [ABSTRACT FROM AUTHOR]

This study aimed to investigate the recent trends of antibiotic resistance (AR) prevalence in Staphylococcus aureus isolated from the milk of animals with clinical mastitis in areas of the Abruzzo and Molise regions in Central Italy. Fifty-four S. aureus isolates were obtained from routine testing for clinical mastitis agents carried out in the author institution in the years 2021 and 2022 and were analyzed for phenotypic resistance to eight antibiotics recommended for testing by European norms and belonging to the antibiotic classes used for mastitis treatment in milk-producing animals. Moreover, the presence of 14 transferable genetic determinants encoding resistance to the same antibiotics was analyzed using qPCR tests developed in this study. Phenotypic resistance to non-β-lactams was infrequent, with only one 2022 isolate resistant to clindamycin. However, resistance to the β-lactam cefoxitin at concentrations just above the threshold of 4 µg/mL was observed in 59.2% of isolates in both years, making these isolates classifiable as methicillin-resistant. The AR genotypes detected were the blaZ gene (50% of 2021 isolates and 44.4% of 2022 isolates), aphA3-blaZ- ermC/T (one 2021 isolate), aphA3-ant6-blaZ-ermC/T (one 2021 isolate), blaZ-ermB (one 2022 isolate) and mecA-mph (one 2022 isolate). An inquiry into the veterinarians who provided the samples, regarding the antimicrobials prescribed for mastitis treatment and criteria of usage, indicated a possible causal relation with the AR test results. The occurrence of AR genotypes did not increase in time, most probably reflecting how mastitis was treated and prevented in farms. However, the frequently observed cefoxitin resistance needs to be explained genotypically, further monitored and limited by modifying antibiotic usage practices. The identification of a mecA-positive isolate in 2022 suggests further investigation if this genotype is emerging locally. [ABSTRACT FROM AUTHOR]

Simple Summary: Dairy cows are commonly affected by mastitis, which causes huge economic losses to the dairy industry worldwide. This work aimed to study this disease by comparing multiple biological specimens (feces, serum, and urine) from individuals with or without clinical mastitis. Integrated analysis of metabolome changes across several key biofluids could facilitate knowledge discovery and reliable identification of potential biomarkers, which could in turn be used to shed light on the early diagnosis of dairy cow mastitis in its subclinical form. To the best of our knowledge, this is the first work dedicated to providing quantitative information on the multi-matrix metabolome of dairy cows with/without clinical mastitis by untargeted 1H-NMR. Several pathways related to energy metabolism and amino acid metabolism have been highlighted to explain the mechanisms of the changes in the dairy cow metabolome related to clinical mastitis. Our findings are the first to provide an integrated view and better understanding of the metabolic mechanism of clinical mastitis, useful for the development of regulated strategies to improve the metabolic status of mastitic dairy cows. Due to huge economic losses to the dairy industry worldwide, mastitis can be considered as one of the most common diseases in dairy cows. This work aimed to study this disease by comparing multiple biological specimens (feces, serum, and urine) from individuals with or without clinical mastitis. This was performed by a single analytical platform, namely 1H-NMR, through a multi-matrix strategy. Thanks to the high reproducibility of 1H-NMR, we could characterize 120 molecules across dairy cow feces, serum, and urine. Among them, 23 molecules were in common across the three biofluids. By integrating the results of multi-matrix metabolomics, several pathways pertaining to energy metabolism and amino acid metabolism appeared to be affected by clinical mastitis. The present work wished to deepen the understanding of dairy cow mastitis in its clinical form. Simultaneous analysis of metabolome changes across several key biofluids could facilitate knowledge discovery and the reliable identification of potential biomarkers, which could be, in turn, used to shed light on the early diagnosis of dairy cow mastitis in its subclinical form. [ABSTRACT FROM AUTHOR]

Hydrogen sulfide (H2S), as an endogenous gaseous signaling molecule, plays an important role in the inflammatory process. Our previous study found that Cystathionine-γ-lyase (CTH) and H2S are correlated with the occurrence and development of Clinical Mastitis (CM) in Holstein cows. However, the functions and regulatory mechanisms of CTH/H2S are still unknown. In this study, the inflammatory mammary cell model based on the MAC-T cell line was established by Lipopolysaccharide (LPS)-induced manner to further explore the function and regulatory mechanism of CTH/H2S in cows with CM. In the inflammatory MAC-T cell, the CTH expression and H2S production were both repressed in an LPS-dose dependent manner, which demonstrated that CTH/H2S is related to the progression of inflammation. The inhibition of CTH/H2S using a selective CTH inhibitor, β-cyano-l-Alanine (BCA), promoted LPS-induced inflammation response and the expression of inflammatory cytokines. However, this was reversed by the H2S donor NaHS, demonstrating that H2S can protect cells from inflammatory damage. Intriguingly, interleukin-8 (IL-8) showed an inverse expression pattern correlated with the H2S-mediated cell protection effect during the inflammation process, and the inhibition test using a selective IL-8 receptor antagonist, SB225002, showed that IL-8 signaling plays a critical role in mediating endogenous H2S synthesis, and CTH/H2S exerts its anti-inflammation via IL-8-mediated signaling. This study provided support for the prevention and treatment of CM and the development of a novel anti-inflammatory strategy. [ABSTRACT FROM AUTHOR]

Toll-like receptor 2 (TLR2) plays a crucial role in bacterial recognition and the host immune response during infection. However, its function and downstream biological processes (BPs) in the mammary glands (MGs) of Holstein cows with clinical mastitis (CM) are not fully understood. This study aimed to comprehensively identify the BPs and differentially expressed proteins (DEPs) associated with the bacterial response and TLR2 using data-independent acquisition (DIA) proteomic data. A possible mechanism for the action of TLR2 was proposed, and the results suggested that the expression levels of TLR2 and caspase 8 (CASP8) were positively correlated with the apoptosis of MGs. The expression patterns of TLR2 and TEK receptor tyrosine kinase 2 (Tie2) were negatively correlated with angiogenesis. These results indicated that TLR2 might promote apoptosis in mammary epithelial cells (MECs) and vascular endothelial cells (VECs) via upregulation of CASP8 expression, and inhibition of angiogenesis in VECs via downregulation of Tie2 expression in dairy cows with CM. In conclusion, TLR2 is associated with inflammation, apoptosis, and angiogenesis in the MGs of dairy cows with bacteria-induced mastitis. These results contribute to a deeper understanding of the pathogenic mechanisms and provide the knowledge needed for developing the prevention and treatment of dairy mastitis. [ABSTRACT FROM AUTHOR]

Simple Summary: Mid-infrared (MIR) spectroscopy is the method of choice to determine milk components like fat, protein and urea. We examined the potential of MIR spectra analyses for the prediction of clinical mastitis events of dairy cows additionally, or alternatively, to somatic cell count, which is routinely used as an indicator for mastitis monitoring. Prediction models based on MIR spectra and a somatic cell count-derived score (SCS) were developed and compared. A model based on MIR spectra and SCS proved more accurate at predicting mastitis than models based on either indicator alone. Consequently, MIR spectra analyses add extra value in the prediction of clinical mastitis, making them potentially useful for dairy farm management and as an auxiliary trait for the genetic evaluation of udder health. Monitoring for mastitis on dairy farms is of particular importance, as it is one of the most prevalent bovine diseases. A commonly used indicator for mastitis monitoring is somatic cell count. A supplementary tool to predict mastitis risk may be mid-infrared (MIR) spectroscopy of milk. Because bovine health status can affect milk composition, this technique is already routinely used to determine standard milk components. The aim of the present study was to compare the performance of models to predict clinical mastitis based on MIR spectral data and/or somatic cell count score (SCS), and to explore differences of prediction accuracies for acute and chronic clinical mastitis diagnoses. Test-day data of the routine Austrian milk recording system and diagnosis data of its health monitoring, from 59,002 cows of the breeds Fleckvieh (dual purpose Simmental), Holstein Friesian and Brown Swiss, were used. Test-day records within 21 days before and 21 days after a mastitis diagnosis were defined as mastitis cases. Three different models (MIR, SCS, MIR + SCS) were compared, applying Partial Least Squares Discriminant Analysis. Results of external validation in the overall time window (−/+21 days) showed area under receiver operating characteristic curves (AUC) of 0.70 when based only on MIR, 0.72 when based only on SCS, and 0.76 when based on both. Considering as mastitis cases only the test-day records within 7 days after mastitis diagnosis, the corresponding areas under the curve were 0.77, 0.83 and 0.85. Hence, the model combining MIR spectral data and SCS was performing best. Mastitis probabilities derived from the prediction models are potentially valuable for routine mastitis monitoring for farmers, as well as for the genetic evaluation of the trait udder health. [ABSTRACT FROM AUTHOR]

Simple Summary: Endogenous hydrogen sulfide (H2S) has been implicated in many physiological and pathological processes, particularly in inflammatory responses and adaptive immunity. In this study, we identified the candidate differentially expressed proteins (DEPs) associated with H2S metabolism in Holstein cows with clinical mastitis (CM). The results revealed 17 DEPs included in 44 Gene Ontology (GO) terms and five Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways related to sulfide metabolism and indicated the important role of cystathionine-γ-lyase (CTH)/H2S in CM. Our findings can support research into the function and regulatory mechanism of CTH/H2S in Holstein cows and provide a basis for the prevention and treatment of CM. H2S plays an important role in various inflammatory diseases. However, the role of H2S and synthetic enzymes in Holstein cows with CM is unknown. The aim of this study was to identify DEPs associated with sulfide metabolism and further investigate their roles in dairy cows with CM. From 3739 DEPs generated by data-independent acquisition proteomics, we identified a total of 17 DEPs included in 44 GO terms and five KEGG pathways related to sulfide metabolism, including CTH and cystathionine-β-synthase (CBS). Immunohistochemical and immunofluorescence staining results showed that CTH and CBS proteins were present mainly in the cytoplasm of mammary epithelial cells. Endogenous H2S production in the serum of the CM group was significantly lower than that of the healthy Holstein cows. CTH and CBS mRNA and protein levels in the mammary glands of the CM group were significantly downregulated compared to those of the healthy group. These results indicate that CTH and H2S were correlated with the occurrence and development of CM in Holstein cows, which provides important insights into the function and regulatory mechanism of CTH/H2S in Holstein cows. [ABSTRACT FROM AUTHOR]

Staphylococcus aureus is a contagious pathogen frequently associated with bovine mastitis in Brazil. Molecular characterization of Staph. aureus isolated from affected mammary quarters of cows with clinical mastitis (CM) can provide data on epidemiological behavior of this pathogen and antimicrobial susceptibility (AMS) assessment at the genotypic level. This study genotypically characterized Staph. aureus isolates recovered from cows with CM and determined the association of genotypes and AMS. A total of 84 Staph. aureus strains identified from affected mammary quarters of cows with CM in 13 dairy herds from Southeastern Brazil were submitted for susceptibility testing to 10 antimicrobials using the technique of minimal inhibitory concentration. The same isolates were also genotyped using the spa-typing methodology. Results showed a high genotypic similarity between the Staph. aureus isolates within and between herds, which were categorized as resistant to most antimicrobials, especially to β-lactam antibiotics. In addition, differences in AMS were observed among genotypic clusters, which may affect the efficacy of antimicrobials used to treat CM in different dairy herds. [ABSTRACT FROM AUTHOR]

When cows on dairy farms are milked with an automatic milking system or in high capacity milking parlors, clinical mastitis (CM) cannot be adequately detected without sensors. The objective of this paper is to describe the performance demands of sensor systems to detect CM and evaluats the current performance of these sensor systems. Several detection models based on different sensors were studied in the past. When evaluating these models, three factors are important: performance (in terms of sensitivity and specificity), the time window and the similarity of the study data with real farm data. A CM detection system should offer at least a sensitivity of 80% and a specificity of 99%. The time window should not be longer than 48 hours and study circumstances should be as similar to practical farm circumstances as possible. The study design should comprise more than one farm for data collection. Since 1992, 16 peer-reviewed papers have been published with a description and evaluation of CM detection models. There is a large variation in the use of sensors and algorithms. All this makes these results not very comparable. There is a also large difference in performance between the detection models and also a large variation in time windows used and little similarity between study data. Therefore, it is difficult to compare the overall performance of the different CM detection models. The sensitivity and specificity found in the different studies could, for a large part, be explained in differences in the used time window. None of the described studies satisfied the demands for CM detection models. [ABSTRACT FROM AUTHOR]

The objective of this study was to compare immediate intramammary antimicrobial treatment of mild and moderate cases of clinical mastitis (CM) with a selective treatment protocol based on on-farm culture results using Accumast®. The study was conducted at a 2600 cow commercial farm in Northeast Germany. Using a randomized design, mild and moderate clinical mastitis cases were assigned to either the blanket therapy (BT) or pathogen-based therapy (SELECT) group. Overall, 468 cases were used for final analyses (BT = 236; SELECT = 232). The percentage of cases assigned to the blanket and pathogen-based groups that received intramammary therapy were 100 and 69.9%, respectively. Implementation of a pathogen-based treatment protocol for mild and moderate CM cases resulted in no significant difference in post-event milk production, somatic cell count, survival to 30 d, and days spent in the hospital compared with a blanket therapy protocol. Cows in the SELECT group had reduced odds of being culled within 60 d post CM (odds ratio = 0.54; 95% CI = 0.31–0.93; p = 0.027). The use of a pathogen-based treatment protocol using an on-farm culture system has the potential to efficiently reduce antimicrobial use without negative effects on health. [ABSTRACT FROM AUTHOR]

Clinical mastitis (CM) is the most common microbial disease treated in dairy cows. We analyzed the antimicrobial usage in cows with CM (n = 11,420) in large dairy herds (n = 43) in Estonia. CM treatment data were collected during a 12-month study period. The antimicrobial usage was observed during the 21 days from the initiation of treatment, and the incidence of antimicrobial-treated CM was calculated for each study herd. The effect of intramammary (IMM), systemic, and combined (systemic and IMM) penicillin treatment of CM on the post-treatment somatic cell count (SCC) was analyzed using the treatment records of 2222 cows from 24 herds with a mixed multivariable linear regression model. The median incidence of antimicrobial-treated CM was 35.8 per 100 cow-years. Procaine benzylpenicillin and marbofloxacin were used in 6103 (35.5%, 95% CI 34.8–36.2) and 2839 (16.5%, 95% CI 16.0–17.1) CM treatments, respectively. Post-treatment SCC was higher after IMM penicillin therapy compared to systemic or combination therapy. Treatment of CM usually included first-choice antimicrobials, but different antimicrobial combinations were also widely used. The effect of procaine benzylpenicillin to post-treatment SCC was dependent on the administration route, cow parity, and days in milk. Further studies should evaluate the factors affecting veterinarians' choice of antimicrobial used in the treatment of CM. [ABSTRACT FROM AUTHOR]

Staphylococcus aureus (S. aureus) is an important pathogen that causes clinical mastitis in goats and produces infections difficult to cure. Different antimicrobials as fluoroquinolones have been used against S. aureus. However, the studies developed to evaluate the bacterial drug interaction only have used the MIC as a single reference point with artificial growth media. The aims of this study were to describe the effect of marbofloxacin on S. aureus isolated from mastitis goats' milk by different approaches as the minimum inhibitory and bactericidal concentrations (MIC and MBC) in cation adjusted Mueller–Hinton broth (CAMHB), serum and milk of goats at two inoculum sizes of 105 and 108 CFU/mL, the determination and analysis of the time kill curves (TKC) by non-linear mixed effect models in each growth medium and inoculum size, as well as the estimation of their pharmacokinetics/pharmacodynamics (PK/PD) cutoff values. The results obtained indicate that MIC values were higher and increases 2,4-fold in serum and 3,6-fold in milk at high inoculum, as well as the EC50 values determined by each pharmacodynamics model. Finally, the PK/PD cutoff values defined as fAUC24/MIC ratios to achieve clinical efficacy were highly dependent on inoculum and growth medium, with median values of 60–180, especially at high inoculum in milk, suggesting that further studies are necessary to evaluate and optimize the best therapeutic strategies for treating S. aureus in lactating goats. [ABSTRACT FROM AUTHOR]

Simple Summary: Bovine clinical mastitis is an inflammatory disease of the mammary gland associated with visual changes in the milk and/or the udder. We show that the incidence of clinical mastitis in commercial dairy farms in Bangladesh is high but with large variation between farms. Streptococci and non-aureus Staphylococci were the most frequently isolated bacteria from quarter milk samples. Staphylococcus aureus and non-aureus Staphylococci were often resistant against penicillin and oxacillin. This work suggests an urgent need for improved udder health management and specifically a more prudent use of antimicrobial agents following a treatment protocol. Clinical mastitis (CM) is an important production disease in dairy cows, but much of the knowledge required to effectively control CM is lacking, specifically in low-income countries where most farms are small and have specific dairy management, such as regular udder cleaning and practicing hand milking. Therefore, we conducted a 6-month-long cohort study to (a) estimate the incidence rate of clinical mastitis (IRCM) at the cow and quarter level, (b) identify risk factors for the occurrence of CM, (c) describe the etiology of CM, and (d) quantify antimicrobial susceptibility (AMS) against commonly used antimicrobial agents in S. aureus and non-aureus Staphylococcus spp. (NAS) in dairy farms in the Chattogram region of Bangladesh. On 24 farms, all cows were monitored for CM during a 6-month period. Cases of CM were identified by trained farmers and milk samples were collected aseptically before administering any antimicrobial therapy. In total, 1383 lactating cows were enrolled, which totaled 446 cow-years at risk. During the study period, 196 new cases of CM occurred, resulting in an estimated crude IRCM of 43.9 cases per 100 cow-years, though this varied substantially between farms. Among the tested CM quarter samples, Streptococci (22.9%) followed by non-aureus staphylococci (20.3%) were the most frequently isolated pathogens and resistance of S. aureus and NAS against penicillin (2 out of 3 and 27 out of 39 isolates, respectively) and oxacillin (2 out of 3 and 38 out of 39 isolates, respectively) was common. The IRCM was associated with a high milk yield, 28 to 90 days in milk, and a higher body condition score. Our results show that there is substantial room for udder health improvement on most farms. [ABSTRACT FROM AUTHOR]

Simple Summary: Bovine mastitis is a complex disease with many potential risk factors. However, few studies have reported the risk factors for mastitis in Brazil. This prospective, repeated cross-sectional study was carried out between May 2018 and June 2019 in smallholder dairy farms located in the southeast of Brazil. The potential risk factors for clinical and subclinical mastitis in smallholder dairy farms in Brazil were investigated by a combination of a questionnaire, clinical udder examinations and tests for subclinical mastitis. Risk factors for mastitis were evaluated at the cow level and at the herd level. The risk factors evaluated at the herd level were related to milking management, environment and management practices. We identified some risk factors; increased parity, later stage of lactation, not milking clinical and subclinical cases last, lack of routine cleaning of the milking parlor, using the dry-off treatment and optimized feed before calving. Further research on more farms across more areas of Brazil is required to develop targeted control programs for mastitis on smallholder dairy farms. The aim of this study was to investigate the potential risk factors for clinical and subclinical mastitis in smallholder dairy farms in Brazil. A prospective, repeated cross-sectional study was carried out between May 2018 and June 2019 on 10 smallholder dairy farms. Potential risk factors for subclinical and clinical mastitis at the herd and cow level were recorded through interviewing the owner and by observation. A combination of clinical udder examination and the Tamis (screened mug with a dark base) test (Tadabras Indústria e Comércio de Produtos Agrovetereinário LTDA, Bragrança Paulista, SP, Brazil) were applied to observe clinical mastitis, and the California Mastitis Test (Tadabras Indústria e Comércio de Produtos Agrovetereinário LTDA, Bragrança Paulista, SP, Brazil) was used to determine subclinical mastitis. A total of 4567 quarters were tested, 107 (2.3%) had clinical mastitis, while 1519 (33.2%) had subclinical mastitis. At the cow level, clinical mastitis risk was highest in mid-lactation (50–150 days in milk) with OR 2.62 with 95% confidence interval (CI) of 1.03–6.67, while subclinical mastitis was highest in late lactation (> 150 days in milk) with OR 2.74 (95% CI 2.05–3.63) and lower in primiparous (OR 0.54, 95% CI 0.41–0.71) than multiparous cows. At the herd level, using dry-cow treatment (OR 4.23, 95% CI 1.42–12.62) was associated with an increased risk of clinical mastitis. Milking clinical (OR 0.37, 95% CI 0.24–0.56) and subclinical cases last (OR 0.21, 95% CI 0.09–0.47) and cleaning the milking parlor regularly (OR 0.27, 95% CI 0.15–0.46) had decreased odds for subclinical mastitis, while herds with optimized feed had greater odds (OR 9.11, 95% CI 2.59–31.9). Prevalence of clinical mastitis was at its lowest at the first visit in June/July and highest at the last visit in April/June (OR 3.81, 95% CI 1.93–7.52). Subclinical mastitis also presented increased odds in the last visit (OR 2.62, 95% CI 2.0–3.36). This study has identified some risk factors for mastitis on smallholder farms but further research on more farms across more areas of Brazil is required to develop a targeted mastitis control program for smallholder farms. [ABSTRACT FROM AUTHOR]

Streptococcus uberis, an environmental pathogen responsible also for contagious transmission, has been increasingly implicated in clinical mastitis (CM) cases in Europe. We described a 4-month epidemiological investigation of Strep. uberis CM cases in an Italian dairy farm. We determined molecular characteristics and phenotypic antimicrobial resistance of 71 Strep. uberis isolates from dairy cows with CM. Genotypic variability was investigated via multiplex PCR of housekeeping and virulence genes, and by RAPD-PCR typing. Antimicrobial susceptibility was assessed for 14 antimicrobials by MIC assay. All the isolates carried the 11 genes investigated. At 90% similarity, two distinct clusters, grouping 69 of the 71 isolates, were detected in the dendrogram derived from the primer ERIC1. The predominant cluster I could be separated into two subclusters, containing 38 and 14 isolates, respectively. Strep. uberis strains belonging to the same RAPD pattern differed in their resistance profiles. Most (97.2%) of them were resistant to at least one of the drugs tested, but only 25.4% showed a multidrug resistance phenotype. The highest resistance rate was observed for lincomycin (93%), followed by tetracycline (85.9%). This study confirmed a low prevalence of β-lactam resistance in Strep. uberis, with only one isolate showing resistance to six antimicrobial classes, including cephalosporins. [ABSTRACT FROM AUTHOR]

Simple Summary: Internal teat sealant (ITS) at drying off is a strategy used for the prevention of intramammary infections (IMI) during the dry period (DP), as it simulates the keratin plug's purpose, which is to prevent the access of pathogens into the mammary gland. The results from randomized clinical trials provide valuable information about the efficacy of commercially available products to be used in dairy cattle, assisting farmers to choose the best protocols for mastitis control. This study evaluated a new ITS infused at drying off as an alternative to prevent IMI during DP in a tropical country. Our results showed no effect of treatments on risk of bacteriological cure, subclinical mastitis (SCM) cure, and new cases of SCM postpartum. On the other hand, the use of ITS combined with an intramammary antibiotic (SDCT) reduced the risk of clinical mastitis up to 60 days postpartum, the overall risk of new intramammary infections (NIMI), and the NIMI caused by major pathogens compared to the use of antimicrobial alone (ADCT). Thus, the use of ITS combined with an antimicrobial at drying off was effective to prevent NIMI and clinical mastitis up to 60 days postpartum. The effect of an internal teat sealant (ITS) on subsequent infection of the mammary gland was evaluated on the following mammary gland health indicators: (a) bacteriological cure of preexisting intramammary infections at drying off, (b) risk of postpartum new intramammary infections (NIMI), (c) cure and risk of new cases of subclinical mastitis (SCM), and (d) risk of postpartum clinical mastitis (CM). A total of 553 cows during late gestation were randomly assigned into two treatment protocols at drying off: (a) Dry cow therapy with 0.25 g of intramammary anhydrous cefalonium (ADCT; Cepravin®, MSD Animal Health); or (b) ADCT combined with ITS (SDCT; 4 g bismuth subnitrate; Masti-Seal®, MSD Animal Health, São Paulo, Brazil). Mammary quarter (MQ) milk samples were collected for microbiological culture and somatic cell count (SCC) at drying off and early lactation, and data from 1756 MQ were used in the multivariate logistic regression. There was no effect on the risk of bacteriological cure, SCM cure, and new cases of postpartum SCM. Still, SDCT reduced the risk of CM up to 60 days postpartum (DPP), overall NIMI risk, and the NIMI caused by major pathogens compared to ADCT. Thus, the DCT combined with ITS at drying off is effective for preventing NIMI during the dry period and CM up to 60 DPP. [ABSTRACT FROM AUTHOR]