Search Results (207)

The objective of this study was to evaluate the effects of feeding a combination of chicory–plantain silage and supplementing Se yeast on the response of early-lactating ewes to induce subclinical mastitis. Polypay ewes (n = 32) were fed either chicory–plantain silage or grass silage and supplemented with 3.6 mg Se yeast/ewe/day for approximately 2 months prior to the infusion of S. uberis into both mammary glands (i.e., intramammary infection or IMI). The ewes had a typical subclinical mastitis response with an 8-fold increase in milk somatic cell count within 24 h post-IMI, a decrease in milk yield, and changes in all milk components measured. The ewes experienced a mild systemic inflammation post-IMI as determined by an increase in rectal temperature and decrease in feed and water intake and, in blood, by an increase in the concentration of ceruloplasmin, haptoglobin, and myeloperoxidase and a decrease in paraoxonase, Zn, advanced oxidation protein products, and hematocrit with no effect on pro-inflammatory cytokines. No effect of silage type, likely due to a low concentration of secondary compounds, or Se supplementation was detected in response to IMI. In summary, the subclinical mastitis model used was effective in mounting an inflammatory response, although this was mild; however, feeding chicory–plantain silage with a low concentration of secondary compounds and supplementing Se yeast had no significant effect on the response of ewes to mammary infection. Full article

Water buffalo (Bubalus bubalis) are a primary source of milk in Pakistan, where bovine mastitis is a significant health issue among cattle, leading to substantial economic losses. Staphylococcus aureus is a predominant pathogen associated with mastitis; however, a detailed molecular characterization of the strains in the country remains limited. We previously characterized mastitis strains from the Hazara division of Khyber Pakhtunkhwa, Pakistan. In this study, we investigated mastitis cases in the Peshawar division, including samples from both animals and human farm workers for comparison. Higher rates of mastitis (67.27% of animals) and sub-clinical mastitis (91.03% of positive animals) were identified in Peshawar than for those (34.55% and 75.31%, respectively) previously observed in Hazara. Methicillin-susceptible S. aureus (MSSA) belonging to clonal complex 9 (ST2454) were predominant. Methicillin-resistant S. aureus (MRSA) belonging to ST22 and ST8 were also detected in the Nowshera district. While no S. aureus colonization was observed among animal handlers, evidence of hand contamination suggests a potential route for pathogen spread. Low levels of antibiotic resistance were noted amongst isolates, but higher rates were seen in MRSA. This study presents only the second comprehensive molecular investigation of S. aureus isolated from buffalo mastitis in Pakistan and indicates a concerning rise in mastitis within the province. Full article

Subclinical mastitis causes economic losses due to reduced milk yield and elevated somatic cell counts (SCCs), despite no visible clinical signs. A higher incidence of subclinical mastitis has been reported in cattle infected with bovine leukemia virus (BLV). Levamisole (LMS), known for its immunomodulatory properties, has been suggested as a potential alternative to antibiotics for mastitis treatment; however, its efficacy in BLV-infected cows, particularly in relation to proviral load (PVL), remains unclear. This study aimed to evaluate the therapeutic effect of LMS on subclinical mastitis and its impact on milk immune responses by classifying BLV-infected cows based on PVL. A total of 42 dairy cows with subclinical mastitis (48 quarters) were grouped as BLV-negative, low-PVL, or high-PVL using a PVL cut-off value of 17.8 copies/10 ng DNA, and were administered LMS orally. Changes in viable bacterial counts, SCCs, and milk leukocyte populations were compared. LMS administration significantly reduced the SCC and milk macrophage numbers, especially in BLV-negative and low-PVL cows. These results suggest that LMS may improve subclinical mastitis in certain BLV-infected cows and that PVL may serve as a useful indicator for treatment responsiveness. However, the limited effect in high-PVL cows and the small sample size have limitations, warranting further investigation. Full article

Milk is frequently susceptible to contamination by potential pathogens, posing risks to both food safety and public health. Cheesemaking often relies on raw milk, where microbial communities—including Staphylococcus xylosus—can play a dual role: (i) contributing to fermentation and (ii) acting as opportunistic pathogens that can be often present in subclinical mastitis and be subjected to carry over in dairy products. In this study, Staphylococcus xylosus was isolated from raw bovine milk (preclinical mastitis) and identified via matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (Biotyper scores: 1.87–2.19). Its susceptibility to erythromycin and to an essential oil blend composed of Myrtus communis, Salvia officinalis, and Cistus ladanifer was evaluated. The essential oil blend produced inhibition zones ranging from 9 mm to 13.3 mm, indicating moderate antibacterial activity. Further Minimum Inhibitory Concentration analysis revealed that Myrtus communis, Salvia officinalis, and the essential oil blend inhibited Staphylococcus xylosus growth at concentrations between 0.5 and 0.25 percent, while Cistus ladanifer required higher levels (1 to 0.5 percent). These findings suggest that selected essential oils—especially in combination—hold promise as complementary antimicrobial agents in food safety and antimicrobial resistance mitigation efforts. Full article

Subclinical mastitis (SCM) is a stealthy but devastating challenge in the dairy industry, leading to economic losses and hindering efforts to achieve milk self-sufficiency. This study investigated the prevalence of SCM, antimicrobial resistance, and virulence profiles of Escherichia coli. A total of 174 milk samples were analyzed using the California mastitis test (CMT), somatic cell counts (SCCs), bacteriological culture, MALDI-TOF MS, and polymerase chain reaction (PCR). The findings revealed that the SCM prevalence was 68/174 (39.08%) based on CMT and SCC. Among SCM-positive samples, 60/68 (88.23%) were identified as E. coli, confirmed by MALDI-TOF MS and PCR assay. The most frequently detected serogroups were 0113 (11.6%) and 0113 (3.3%). Additionally, the genes for Stx1 and Stx2 were also detected in nine (15%) and one (1.7%), respectively. Antimicrobial susceptibility tests showed widespread resistance, with E. coli isolates demonstrating resistance to penicillin in 43 (71.6%), followed by ciprofloxacin in 42 (70%) and gentamicin in 18 (30%). A larger proportion of the E. coli strains (100%) harbored the bla_VIM gene, while 23 (38.3%), 20%, 20%, and 1.47% contained bla_KPC, bla_NMD, suli1, and msrA. Thirty (50%) isolates were considered multidrug-resistant (MDR). These findings underscore the urgent need for enhanced surveillance and antibiotic stewardship in dairy farming. The presence of MDR E. coli in SCM poses a dual threat of potential transmission to humans and treatment failures in mastitis management. This study highlights the importance of proactive control strategies to mitigate the spread of antimicrobial resistance in livestock and beyond. Full article

Subclinical mastitis is a critical disease affecting camel health and milk quality. However, research on shifts in milk bacterial communities following subclinical mastitis in camels is limited. We evaluated changes in bacterial communities following subclinical mastitis in Bactrian camels. Three portions of California Mastitis Test (CMT)-negative milk and five portions of CMT-positive milk were collected from each Jimunai County and Keping County using the CMT, and the bacterial community composition of the camel milk was analyzed using amplicon sequencing of the v34 region of the 16S rRNA gene. Subclinical mastitis induced genus-level differences in the core bacterial microbiota of Bactrian camel milk. To our knowledge, Delftia was identified in camel milk for the first time, predominantly in Jimunai County. Bacterial abundance in camel milk from Keping County was increased and altered. Alpha diversity analysis revealed that subclinical mastitis induced lower and higher bacterial abundance in milk from Jimunai County and Keping County, respectively, compared to that of healthy camels. Therefore, these findings provide direction for future research on pathogenic microorganisms for the prevention and control of subclinical mastitis in Bactrian camels. Full article

Mastitis is a major issue in dairy cows, with subclinical mastitis (SCM) being hard to detect and potentially progressing to clinical mastitis. Antibiotic use raises concerns about resistance and milk contamination, highlighting the need for natural alternatives. Sodium alginate (SA), known for its antioxidant and immunomodulatory properties, may offer a solution, though its effects on mastitis are unclear. Intramammary infusion of 1% SA (30 mL) was tested in both healthy cows (n = 8; somatic cell count, SCC ≤ 100,000 cells/mL) and those with SCM (n = 12; SCC ≥ 200,000 cells/mL). The results showed that SA significantly increased SCC in both healthy and SCM cows, with peak levels at 48 h, returning to baseline levels thereafter. In cows with SCM, SA treatment led to a 58.3% cytological and 54.5% bacteriological cure rate after 14 days. Additionally, significant downregulation was observed in tumor necrosis factor (TNF)-α, interleukin (IL)-1β, IL-2, IL-4, IL-6, and interferon (IFN)-γ. Conversely, the levels of IL-8, IL-10, and IL-12 initially increased, then declined gradually. Importantly, there were no significant effects on milk composition. These findings suggest that SA may offer an alternative to antibiotics, aiding in immune response and bacterial clearance without the risk of antibiotic residues, thus preventing SCM progression to clinical mastitis. Full article

The 2024 outbreak of highly pathogenic avian influenza virus (HPAIV) H5N1 in U.S. dairy cattle presented an unprecedented scenario where the virus infected bovine mammary glands and was detected in milk, raising serious concerns for public health and the dairy industry. Unlike previously described subclinical influenza A virus (IAV) infections in cattle, H5N1 infection induced severe clinical symptoms, including respiratory distress, mastitis, and abnormal milk production. To understand the host immune responses and changes, particularly in the mammary gland, we performed single-cell RNA sequencing analysis on bovine milk somatic cells (bMSCs) in vitro exposed to an H5N1 isolate from an infected dairy farm. We identified ten distinct cell clusters and observed a shift toward type-2 immune responses, characterized by T cells expressing IL13 and GATA3, and three different subtypes of epithelial cells based on the expression of genes associated with milk production. Our study revealed temporal dynamics in cytokine expression, with a rapid decline in luminal epithelial cells and an increase in macrophages and dendritic cells, suggesting a role in increased antigen presentation. While viral RNA was detected in bulk-exposed bMSC samples via qRT-PCR, no viral reads were observed in the scRNA-seq data, indicating that the immune responses captured may be due to exposure to viral components rather than productive infection. This research fills a critical gap in understanding the immune responses of bovine mammary glands to H5N1 exposure and highlights the need for further investigation into therapeutic strategies for managing such outbreaks. Full article

Subclinical mastitis (SM) is the most economically damaging yet often visually undetectable disease of dairy cows. Early detection and treatment can reduce the loss caused by the disease; thus, the continuous improvement of SM diagnostic methods is necessary. Although milk’s somatic cell count (SCC) is commonly measured for diagnostic purposes, its direct determination is not widely used in everyday practice. The primary objective of our work was to investigate whether the predictive value of SM diagnostics can be improved by training artificial neural networks (ANNs) on data generated using typical conventional milking systems. The best ANN classifier had a sensitivity of 0.54 and a specificity of 0.77, which is comparable to performances of various California Mastitis Tests (CMT) found in the literature. Combining two diagnostic tests, ANN and CMT, we concluded that the positive predictive value could be up to 50% higher than the value provided by the individual CMT. While implementing CMT is a labor-intensive process on herd-level, in milking machines where milk properties or milk yield data can be measured automatically, similar to our work, SCC-increase predictions for all individuals could be obtained daily basis. Full article

This study assessed the impact of farming systems on aflatoxin M1 (AFM1) content and the prevalence of mastitis-causing bacteria in goat milk. A total of 233 milk samples were collected from two Skopelos goat farms—one intensive and one extensive farm—and analyzed for AFM1 content using a competitive ELISA. An additional 219 samples from goats suspected of subclinical mastitis were tested for bacterial prevalence with microbial culturing. The results showed that AFM1 concentration was significantly higher in the intensive farming system (7.76 ± 0.76 ng/kg) than in the extensive farming system (3.78 ± 0.79 ng/kg), though it remained below the legal limit of 50 ng/kg. The main effects of season and year were not significant, though higher levels of AFM1 were observed during winter. The interaction effects of season–farming system and year–season–farming system on AFM1 levels were significant. The prevalence of mastitis-causing bacteria varied by system, with Streptococcus spp. being more common in the extensive farming system, and Staphylococcus aureus was more frequently detected in milk samples from the intensive farming system. Binomial regression indicated that both the farming system and lactation stage significantly influenced Streptococcus spp. prevalence (p = 0.05; OR = 1.9 and 2.7, respectively). It is concluded that the farming system affects those quality parameters in goat milk. Full article

Background/Objectives: The resistance to occult mastitis in dairy cows is a multifaceted trait influenced by a variety of genetic and environmental factors, posing significant challenges to its prevention and treatment. Methods: In this study, a cohort of 389 Holstein dairy cows was selected for investigation. The genes NOD2, CXCR1, SPP1 and LF, which are implicated in resistance to occult mastitis, were genotyped utilizing the efficient and cost-effective Kompetitive Allele-Specific PCR (KASP) technology. Additionally, the study analyzed the association between various single nucleotide polymorphisms (SNPs) and the somatic cell score in Holstein dairy cows. Multi-locus penetrance variance analysis (MPVA) analysis was also conducted to assess the resistance of different genotypic combinations to recessive mastitis in dairy cows. A genotyping kit for occult mastitis resistance was developed. Subsequently, 300 Holstein cows were randomly selected to evaluate the accuracy of the kit’s classification and resistance detection. Results: The findings revealed that the most effective genotype combination was SPP1(AA)-CXCR1(CC)-NOD2(CA)-LF(GA). Upon verification, the genotyping kit for recessive mastitis resistance in dairy cows exhibited an accuracy rate of 100% for individual genotyping and 95.90% for resistance detection. Conclusions: From the perspective of disease resistance genetics, this study lays a foundation for the precise management of dairy cow herds. It enables the early identification and removal of individuals susceptible to subclinical mastitis, thereby improving the overall quality of the cattle population. Full article

The dairy industry relies on the health and well-being of dairy cows for the optimal production of milk and dairy products. Mastitis, a prevalent and economically burdensome disease characterized by udder inflammation, poses significant challenges to dairy farmers worldwide. In this study, we employed peptidomics to explore the peptide profiles of dry secretions collected from dairy cows at specific intervals during the dry-off period. We hypothesized that alterations in peptide composition during the dry period may influence pathogen proliferation and immune cell functioning, thereby impacting mastitis susceptibility. Our objectives were to investigate the following: (i) differences in peptide composition and alterations between healthy cows and those with subclinical mastitis, potentially serving as biomarkers for early mastitis detection and offering insights into udder bioprocesses; (ii) variations in peptide profiles between the early (day 2) and later (day 21) stages of the dry-off period during both health conditions. Dry secretions were collected from 16 udder quarters of 8 cows at two defined time points—Day 2 (D2) and Day 21 (D21)—during the dry period. Our results revealed distinct peptide patterns between healthy and subclinical mastitis cows, as well as temporal variations in peptide profiles throughout the dry-off period. A total of 1235 peptides, originating from 59 distinct proteins (primarily β-casein), were detected across the four groups: subclinical mastitis day 21 (SCM-D21), subclinical mastitis day 2 (SCM-D2), healthy day 21 (H-D21), and healthy day 2 (H-D2). Furthermore, 56 out of the 1235 peptides identified in total matched known functional peptides, with a total of 17 different functions including antimicrobial, antioxidant, and immunomodulatory, suggesting their potential roles in mastitis pathogenesis and mammary gland physiology. Comparative analyses revealed changes in the levels of these functional peptides across the four different groups, suggesting their potential roles in regulating immune responses, oxidative stress, inflammatory processes, and other biological activities during subclinical mastitis and the dry-off period. These findings provide valuable insights into mastitis detection, management strategies, and dairy cow health monitoring, offering promising avenues for enhancing milk quality and dairy industry sustainability. Full article

Mastitis is a major infectious disease that causes significant economic losses in the dairy industry. Current control programs primarily rely on antimicrobials, contributing to the growing concerns of the resistance situation and drug residues in milk. This study aimed to identify the bacterial pathogens responsible for subclinical/clinical bovine mastitis, assess their antimicrobial resistance profiles, and evaluate the antimicrobial effects of Caesalpinia sappan in treating mastitis through both in vitro and in vivo studies. A total of 138 bacterial isolates, representing 40 species, were identified from 100 milk samples collected from dairy cows under the Maejo Cooperative Group in Chiang Mai, Thailand, between May 2021 and February 2022. The most prevalent species was Escherichia coli (10.87%), followed by Bacillus cereus (9.42%) and Staphylococcus sciuri (7.97%). The highest resistance rates were observed for penicillin (65.21%), followed by streptomycin and lincomycin (61.59%). Nine isolates resisted all 18 antimicrobials tested. The average minimum inhibitory concentration (MIC) of C. sappan against the identified pathogens ranged from 0.63 to 17.68 mg/mL, with the highest MIC observed against Pseudomonas luteola. In the animal experiment, treatment with a prototype of an intramammary infusion compound primarily formulated from C. sappan significantly reduced the total bacterial count and California Mastitis Test (CMT) scores (p < 0.01). These results suggest that C. sappan exhibits antimicrobial efficacy against various mastitis bacteria and could serve as a potential alternative treatment to control bovine mastitis in dairy cattle within the study region. Full article

In the present study, we isolated S. canis from milk samples in a dairy farm with suspicions of subclinical mastitis. Milk samples testing positive on California Mastitis Test (CMT) were collected from different cows for bacteriological and genomic analyses to identify the causative pathogen, and somatic cell counts (SCC) were determined. A multiplex qPCR assay was conducted to detect 15 potential pathogens, and all samples showed negative results. Conventional bacteriology procedures were performed; DNA of the bacterial strains was extracted, sequenced, and submitted to bioinformatic analysis. Three CMT positive milk samples showed SCC > 200 × 10³ cell/mL. However, these same three samples were positive for bacteria phenotypically identified as Streptococci, and the strains were confirmed as S. canis using MS MALDI-TOF methodology. In susceptibility testing, resistance against tetracycline was detected, revealing a potential chronic infection in one cow, while the presence of the same bacteria was observed in two other cows. Genomic DNA from four S. canis isolates, obtained in the first and second sampling, was sequenced. Genetic relationships revealed a unique sequence type (ST24). The gene (tetM) related with resistance to TE was highlighted. Although the association between S. canis and mastitis is not routinely detected, early diagnosis of bacterial infections and the study of the antimicrobial profile are crucial for effective therapy. Pets could act as a potential reservoir, so improving hygienic conditions is needed to prevent new infections. Full article

Mastitis, an inflammatory condition of the udder, can be caused by the entry of Staphylococcus aureus, whose adhesion to the mammary epithelial cells is influenced by virulence factors such as microbial surface components recognizing adhesive matrix molecules (MSCRAMMs) and the accessory gene regulator (agr) system. Our goal was to determine the adhesion and invasion rates of S. aureus isolates from clinical (mild and moderate) and subclinical mastitis and to assess the impact of MSCRAMM genes and agr types on disease severity. Clinical isolates predominantly carried agrII (p < 0.0083) and multiple MSCRAMM genes, correlating with high adhesion capacity but reduced invasion capacity regardless of clinical severity. Remarkably, subclinical isolates, mainly agr-negative (85.7%), showed increased cellular invasion (p < 0.0001), possibly due to reduced expression of agr-mediated virulence factors. These findings contribute to the understanding of the pathogen–host dynamics in bovine mastitis and highlight the importance of both MSCRAMMs and the agr system in modulating disease severity. These insights can inform targeted interventions for mastitis prevention and treatment. Full article