Search Results (150)

Reducing antimicrobial use in dairy cattle is essential for limiting antimicrobial resistance while maintaining udder health. Selective dry cow therapy (SDCT), in which antibiotics are administered only to cows at increased risk of intramammary infection, has become an important antimicrobial stewardship strategy, yet information regarding its acceptance in Romania is lacking. This cross-sectional study evaluated the knowledge, perceptions, and acceptance of antimicrobial resistance and SDCT among Romanian dairy farmers and veterinarians using an online questionnaire completed by 132 respondents. The questionnaire assessed current dry-off practices, awareness of antimicrobial resistance and selective dry cow therapy, previous implementation, perceived barriers and facilitators, and willingness to adopt or recommend selective dry cow therapy. Overall, 70.5% of farmers had previously heard of antimicrobial resistance, whereas only 27.9% were familiar with the formal concept of SDCT. Administration of antibiotics only to selected cows at dry-off was already the most commonly reported practice among farmers (41.0%) and veterinarians (47.9%), indicating that selective antibiotic use was already being applied despite limited familiarity with the formal SDCT concept. After receiving standardized information describing the strategy, most respondents expressed favorable attitudes and willingness to implement or recommend selective dry cow therapy. Lack of knowledge, limited availability of herd-level diagnostic information, and concerns regarding mastitis were identified as the principal barriers to implementation. Multivariable analysis demonstrated that concern regarding antimicrobial resistance was the only independent predictor of SDCT acceptance among farmers. These findings suggest that strengthening antimicrobial stewardship education, veterinary guidance, standardized protocols, and access to diagnostic tools may facilitate wider adoption of selective dry cow therapy in Romania. Full article

This review examines the relationship between climate-driven heat stress (HS) and bovine mastitis in the Po Valley, a key European dairy region characterized by intensive production systems and increasing climatic vulnerability. It aims to contextualize how rising temperature–humidity index (THI) levels influence animal health and productivity. This study synthesizes the current literature on biometeorological conditions, epidemiological trends, and physiological mechanisms linking HS to mastitis. Evidence indicates that prolonged exposure to elevated THI impairs thermoregulation, disrupts endocrine and metabolic balance, and weakens immune function, thereby increasing susceptibility to intramammary infections. Epidemiological data reveal a clear seasonal pattern, with mastitis incidence peaking during summer months and a growing predominance of environmental pathogens. Additionally, HS negatively affects milk yield and quality, amplifying economic losses in dairy systems. The findings highlight that mastitis in this context is not merely an infectious disease but a multifactorial condition shaped by environmental, physiological, and management factors. Overall, this review underscores the need for integrated mitigation strategies, including improved housing, nutrition, genetic selection, and precision monitoring, to enhance resilience. In the face of ongoing climate change, adapting dairy production systems will be essential to safeguard animal welfare, maintain productivity, and ensure the long-term sustainability of the Po Valley dairy sector. Full article

Selective dry cow therapy (SDCT) has emerged as a key strategy to reduce antimicrobial use in dairy production while maintaining udder health. This study aimed to evaluate the feasibility and impact of implementing SDCT in Romanian dairy farms by comparing low-risk cows treated with internal teat sealant only (ITS) at dry-off with low-risk cows treated with intramammary antibiotics at dry-off. A prospective field study was conducted on two commercial dairy herds, including 87 cows classified based on somatic cell count (SCC) and differential SCC (DSCC), and compared with a historical cohort of 37 cows. Udder health parameters, milk yield during the first 100 days in milk (DIM), antimicrobial use, and economic outcomes were evaluated. No significant differences were observed between groups in terms of postpartum intramammary infections, somatic cell score, DSCC, or clinical mastitis incidence. Milk yield during early lactation was also not affected by treatment. The ITS-only strategy resulted in a substantial reduction in antimicrobial use (−88.8% per cow) without significant differences in total economic costs. Farm-related differences highlighted the influence of management conditions on outcomes. These findings indicate that, in low-risk cows, SDCT using ITS alone is a safe and effective alternative to antibiotic treatment and support the feasibility of implementing SDCT under Romanian dairy production conditions as a sustainable strategy to promote the targeted and prudent use of antimicrobials while reducing unnecessary antibiotic exposure in dairy herds. However, given the limited number of herds and animals included, further studies are needed to confirm these promising findings under a broader range of production conditions. Full article

The aim of this study was to characterize the coordinated dynamics of the mammary microbiome, antibiotic resistance genes (ARGs), and mobile genetic elements (MGEs) across the dry period, calving, and early lactation. The mammary microbiome undergoes substantial ecological changes across these stages, yet the coordinated dynamics of microbial composition, ARGs, and MGEs remain poorly understood. Here, shotgun metagenomic sequencing was performed on mammary secretion samples collected before dry-off (BM), immediately after calving (ACM), and one month postpartum (AM). The mammary microbiome exhibited a clear “exposure–bottleneck–reassembly” trajectory. BM was characterized by high microbial diversity and the enrichment of environmentally associated taxa, whereas ACM displayed a pronounced immunological bottleneck with markedly reduced microbial diversity and network complexity. During AM, microbial communities partially recovered but remained distinct from the BM state, indicating persistent ecological restructuring after calving. ARGs and MGEs showed parallel dynamics, with broad resistome and mobilome diversity in BM, a sharp contraction in ACM, and a selective re-expansion in AM. Network analysis further revealed maximal ecological complexity in BM, increased ARGs/MGEs connectivity in ACM, and partial stabilization in AM. These findings demonstrate that host physiological transitions, together with dry cow therapy (DCT), drive the coordinated remodeling of the mammary microbiome, resistome, and mobilome across the dry period. Full article

Staphylococcus aureus (S. aureus) bovine mastitis is a significant public health issue. Despite enormous efforts, important gaps remain regarding host–microenvironmental factors. How intramammary reduced oxygen modulates S. aureus transcription in bovine mammary epithelial cells (MECs) remains unclear. We examined oxygen-associated transcriptional changes in a bovine-mammary adapted S. aureus clone following internalization into MECs and identified functional category enrichments under Normal-O₂ and Reduced-O₂ exposures. Bovine MAC-T monolayers were infected with a dominant bovine mastitis isolate under Normal-O₂ or Reduced-O₂ conditions. Triplicate infection experiments were performed for each oxygen condition. Each condition included matched non-reacted bacterial controls maintained under the same gas condition but without MAC-T exposure serving as the reference condition for expression calling. RNA was extracted and profiled using a high-throughput qRT-PCR platform covering genome-wide loci. Expression calls were mapped to curated BioQT roles and interpreted descriptively. Results indicated 211 loci were upregulated and 99 were downregulated under Normal-O₂ conditions, versus 53 upregulated and 35 downregulated under Reduced-O₂ conditions, relative to their non-reacted controls. Under Normal-O₂ conditions, regulated loci covered multiple functional roles, including cellular processes, transport/binding proteins, regulatory functions, and energy metabolism with downregulated loci enriched in transport/binding and cell-envelope categories. Under Reduced-O₂ conditions, upregulated loci were abundant in cellular process annotations dominated by pathogenesis/toxin-related functions, whereas downregulated loci were enriched in nucleotide biosynthetic and DNA/cell division categories. Thus, this reveals oxygen-associated shifts in the transcriptional response of intramammary S. aureus in MAC-T cells. Normal-O₂ conditions were associated with broader category representation, whereas Reduced-O₂ conditions yielded a narrower distribution enriched for selected toxin/pathogenesis- and iron/cation-associated annotations. These oxygen-linked transcriptional-shifts highlight candidate pathways for the intramammary adaptation of S. aureus, potential diagnostic markers, anti-virulence strategies, and targeted therapeutics. Full article

Bovine mastitis is a major infectious disease in dairy cattle, causing significant economic losses and compromising animal health and milk quality worldwide. Among its etiological agents, Staphylococcus aureus is a key contagious pathogen due to its ability to establish persistent intramammary infections and evade host immune responses and antimicrobial therapy. This review summarizes current knowledge on the epidemiology, pathogenesis, clinical presentation, diagnosis, and control of S. aureus in bovine mastitis. Particular emphasis is placed on virulence mechanisms, including adhesion, intracellular persistence, biofilm formation, and immune evasion, which contribute to chronic and recurrent infections. The increasing prevalence of antimicrobial resistance, including methicillin-resistant and multidrug-resistant strains, is highlighted as a major challenge limiting treatment efficacy and posing risks within a One Health context. The review also discusses emerging alternative therapies and innovative control strategies, such as anti-biofilm approaches, immunomodulation, and improved diagnostics, aimed at reducing antimicrobial use. Advances in molecular and point-of-care diagnostic tools are considered for their role in early detection and targeted interventions. Overall, effective control of S. aureus mastitis requires integrated strategies combining prudent antimicrobial use, alternative therapies, improved hygiene, and a multidisciplinary One Health approach. Full article

Enterococcus lactis is increasingly recognized as an emerging mastitis pathogen, yet the functional basis of its virulence and associated health risks remain poorly defined. This study presents an integrated genomic and phenotypic characterization of E. lactis strain EL-A150 isolated from bovine subclinical mastitis. Whole-genome sequencing revealed a 2.49 Mb circular chromosome encoding multiple genes associated with adhesion (acm, bepA, fms, sagA), biofilm formation (empB, empC) and antimicrobial resistance, including determinants related to aminoglycosides and macrolides. Phenotypic assays demonstrated rapid growth, strong biofilm-forming capacity and high adhesion to bovine mammary epithelial cells, while internalization remained low and intracellular persistence was transient. Comparative genomic analyses confirmed the taxonomic placement of the strain within the E. lactis clade (ANI up to 99.5% against reference genomes) and revealed a limited resistome composed of chromosomally encoded genes, with no detectable plasmids or major mobile genetic elements. Collectively, these findings demonstrate that E. lactis EL-A150 possesses a coordinated set of traits conducive to intramammary colonization, supporting its classification as an opportunistic pathogen. The convergence of virulence potential and clinically relevant antimicrobial resistance within a single isolate underscores a One Health concern and highlights the need for surveillance frameworks that integrate functional validation with genomic risk assessment. Full article

Bovine mastitis causes significant economic losses in the dairy industry. Emerging evidence highlights the critical role of long non-coding RNAs (lncRNAs) in inflammation-associated epigenetic regulation through competing endogenous RNA (ceRNA) networks. In this study, we established a bovine mastitis model in three healthy primiparous Holstein cows by intramammary infection with S. aureus. Infected and control mammary tissue samples were then collected for transcriptomic profiling, which identified 2005 differentially expressed lncRNAs. Among them, BMNCR was significantly upregulated in S. aureus-infected mammary tissues and S. aureus-stimulated BMECs. We evaluated the coding potential of BMNCR and confirmed its non-coding nature. Functional studies in BMECs demonstrated that knockdown of BMNCR suppressed proliferation, promoted apoptosis, and altered the expression of inflammatory factors, including IL-2, IL-6, IL-8, and IL-12. Mechanistically, BMNCR acted as a sponge for bta-miR-145, thereby leading to the derepression of ANO6. Silencing ANO6 partly recapitulated the effects of BMNCR knockdown, impairing proliferation and increasing IL-8 expression. Collectively, these findings suggest that the BMNCR/miR-145/ANO6 axis is involved in the regulation of inflammatory responses and epithelial homeostasis during bovine mastitis, with BMNCR functioning as a protective regulator in this process. Full article

Somatic cell count (SCC) in milk is widely used as an indicator of intramammary infections in dairy sheep and is routinely monitored by the dairy industry as a marker of milk quality. This study aimed to evaluate the effect of SCC levels on milk production, composition, colour, coagulation properties, and cheese-making ability in Manchega dairy sheep. A total of 752 individual milk samples were analysed. To normalise SCC distribution, the somatic cell score (SCS) was calculated and samples were classified into SCS classes. Increasing SCS significantly reduced daily milk yield and lactose content, increased milk pH, and decreased lightness (L*). Higher SCS was also associated with impaired coagulation properties, including longer rennet clotting time (RCT) and curd firming rate (k₂₀), as well as reduced curd firmness (A₃₀, A₆₀). Similar effects were observed for modelled coagulation parameters, with delayed RCTeq and reduced kCF and CFp. Regarding cheese-making ability, SCS significantly affected curd humidity and protein recovery, whereas no significant effects were detected for dry curd yield or fat recovery. Overall, elevated somatic cell counts were associated with a reduction in the technological quality of Manchega sheep milk, particularly affecting coagulation behaviour and curd characteristics. These results underline the importance of controlling SCC levels in dairy sheep systems for both udder health monitoring and maintaining milk suitability for cheese-making. Full article

Background/Objectives: Staphylococcus aureus (S. aureus) intramammary infection remains a major global dairy problem due to its contagious nature, its ability to persist and colonize teat/skin and mucosal niches, and the often-limited bacteriological cure achieved with antimicrobial therapy. Beyond udder health, it is relevant to public health because it can enter raw milk chains and serve as a reservoir for antimicrobial resistance determinants that may circulate between dairy animals and humans. Methods: We assessed S. aureus’ ecology in raw ewe milk from 75 sheep farms in Epirus (Greece) by sampling clinically healthy controls (group A) and clinical mastitis cases pre-treatment (group B), followed by resampling at the first post-withdrawal milking after penicillin/streptomycin treatment (group C1—therapeutic protocol 1), oxytetracycline treatment (group C2—therapeutic protocol 2), or enrofloxacin treatment (group C3—therapeutic protocol 3). Results: S. aureus detection was high and comparable across groups (A 23.0%, B 22.0–30.0%, C 20.0–22.0%), and paired analyses showed no significant pre–post shifts in detection/burden within therapeutic protocols (all p > 0.05). Nevertheless, persistence remained evident. The chromosomal gene mecA was detected in S. aureus strains in all groups, ranging from 13.6% in controls to 54.5% post-withdrawal in group C1, and was also present in the pre-treatment group. In paired sampling animals, mecA was mostly stable, with rare emergence or loss. Across antibiotic classes, within-animal resistance transitions were generally uncommon and non-significant (p > 0.05); β-lactam resistance was fully stable (p = 1.00). Descriptively, resistance to protein synthesis inhibitors tended to decline after therapy in protocol 1 and protocol 3, while protocol 3 showed post-treatment gains in fluoroquinolone resistance. By contrast, virulence-associated phenotype traits shifted after therapy: enterotoxigenicity increased post-withdrawal (especially in the C3 group), Staphylococcal Enterotoxin A (SEA) and Staphylococcal Enterotoxin B (SEB) appeared only post-therapy, Staphylococcal Enterotoxin D (SED) increased significantly in paired isolates (p = 0.002), and strong biofilm adherence increased (in C3, p = 1.5 × 10⁻⁵). Conclusions: The detection of S. aureus after therapy suggests that one possibility is that antimicrobial exposure may select for, or otherwise reshape, the residual intramammary population, rather than reliably eliminating it—an outcome that remains clinically relevant for udder health. Moreover, the persistence of mecA/methicillin-resistant Staphylococcus aureus (MRSA)-compatible profiles indicates that milk released to the food chain after withdrawal compliance may still harbor S. aureus with enhanced preservation capacity and significant food safety relevance. Full article

The dry period is a critical window of susceptibility to intramammary infection in dairy cows, yet the metabolic environment of the mammary gland during this phase remains largely uncharacterized. The objective of this study was to profile the metabolome of bovine dry secretion fluid from quarters with subclinical mastitis (SCM; SCC ≥ 200,000 cells/mL) and healthy quarters (H; SCC < 200,000 cells/mL) on day 2 (D2) and day 21 (D21) of the dry period (n = 10 per group quarters per group, drawn from a cohort of 41 enrolled Holstein dairy cows) using high-performance chemical isotope labeling liquid chromatography–mass spectrometry (CIL-LC–MS). A total of 474 metabolites were positively identified. At D2, 186 metabolites differed significantly between SCM and H quarters, with dipeptides dominating the upregulated metabolites, indicating active proteolysis in infected quarters. Norepinephrine was the most significantly depleted metabolite (FC = 0.27, p = 3.37 × 10⁻⁷), pointing to local catecholamine exhaustion. By D21, only 36 metabolites remained altered, representing an 80.6% attenuation of the SCM signature. Interestingly, temporal changes from D2 to D21 far exceeded disease-related differences, with 316 metabolites shifting in both SCM and healthy quarters, establishing mammary involution as the dominant metabolic event during the dry period. Principal component and PLS-DA analyses confirmed that time, not disease status, was the primary driver of metabolic variation. Pathway analysis revealed significant perturbations in amino acid metabolism, glyoxylate and dicarboxylate metabolism, and tryptophan metabolism. These findings provide the first comprehensive metabolomic map of bovine dry secretion, reveal that subclinical mastitis superimposes a proteolytic and neuroimmune disruption onto the involution process, and identify candidate biomarkers for early detection of intramammary infection during the dry period. Full article

Hafnia alvei, which belongs to the Enterobacteriaceae family, has been occasionally documented in animal infections but is still not well characterized in the context of bovine mastitis. This research examined the prevalence and antimicrobial resistance characteristics of H. alvei in dairy cows suffering from subclinical mastitis in South Africa’s Free State Province. In the Thabo Mofutsanyana District, a total of 174 milk samples were obtained from cows on six different dairy farms. The California Mastitis Test (CMT) was used to screen for subclinical mastitis, and somatic cell count was used to confirm it. Standard culture methods were used for bacterial isolation, and presumptive Enterobacteriaceae isolates were identified through matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS). Out of the 174 samples, 84 (48.2%) tested positive for CMT, and 68 (39.1%) met the SCC criteria for subclinical mastitis at a cow level, while 96/336 (28.5%) were infected at a quarter level. Of the 100 presumptive Enterobacteriaceae isolates, 33 (33.0%) were identified as H. alvei (p = 0.0034). Antimicrobial susceptibility testing showed that 50% of the isolates were resistant to penicillin, followed by tetracycline and erythromycin with 25% and 10%, respectively. Furthermore, the results showed that 17 (51.5%) isolates exhibited multidrug-resistant profiles. The results suggest that H. alvei could be a contaminant in raw milk associated with bovine subclinical mastitis in this area, necessitating additional epidemiological research that includes healthy matched controls. Full article

An outbreak of mastitis caused by Staphylococcus aureus occurred in a commercial dairy goat herd during kidding season, resulting in fatal gangrenous mastitis in approximately 30% of the herd. S. aureus was recovered from milk, mammary tissue, and other organs in does subjected to necropsy. The S. aureus milk culture-positive rate among does in the hospital pen was 58.3%, while whole-herd milk cultures of clinically normal mature does identified S. aureus in 15.0% with an additional 15.0% positive for coagulase-negative Staphylococcus (CNS), yielding a total culture-positive rate of 30.0%. The prevalence of CNS in subclinical animals was consistent with previous reports from U.S. dairy goats; in contrast, S. aureus isolation rates substantially exceeded previously reported prevalences. Poor milking hygiene and milking machine dysfunction were identified as major factors contributing to the spread of the S. aureus from goat to goat. California Mastitis Test (CMT) scores were significantly higher in culture-positive does compared with culture-negative animals (p < 0.05), demonstrating the value of CMT as a practical on-farm tool for early treatment decision making. Interventions focused on addressing milking hygiene and milking machine maintenance, as well as segregation and vaccination of replacement females. S. aureus dropped to undetectable in the next two kidding seasons, whereas the CNS culture rates remained unchanged, suggesting other factors may be contributing to CNS infection. This case highlights the role of subclinical intramammary infection and milking practice factors in transmission and control of contagious mastitis pathogens like S. aureus. Full article

Historically, methods to detect subclinical mastitis have been used to aid treatment decisions. The limitations of these tests have led to the exploration of the use of differential somatic cell count (DSCC). The objective of this study was to evaluate the relationship between DSCC and antibiotic success in subclinically infected cows. Cows were pre-screened to confirm an SCC ≥ 200,000 cells/mL and a positive bacterial culture. At enrollment, aseptic quarter milk samples were collected for culturing and evaluated using SCC, DSCC, the California Mastitis Test (CMT), and electrical conductivity. Enrolled quarters were treated with an intramammary antibiotic and resampled 7-, 14-, 28-, and 35-days post-treatment and retrospectively classified as “Cured” or failed to cure (Failed) based on a bacteriological cure. The overall cure rate was 51.5%. Pre-treatment SCC, total leukocyte count (TLC), neutrophils, and CMT were lower in Cured quarters compared to Failed. Meanwhile, macrophage percentage was greater in Cured quarters compared to Failed. No difference was found between lymphocyte percentage and conductivity. When evaluating the use of multiple threshold optimizations to predict cure outcome, TLC tended to have the highest ranking for the metric of interest; however, SCC tended to numerically perform as well in sensitivity and specificity outside of the optimized parameter. Full article

Prevalence of LPS in Gram-negative bacterial udder infections determines mastitis severity and disease prognosis. This pilot study explores the notion that milk-soluble (s) LPS/IgG complex levels in dairy cows link mastitis severity to intramammary Gram-negative infections during early lactation. Milk, within a single herd, was analysed from (i) 34 early lactating cows with acute mastitis and (ii) milk selected from peak lactation cows displaying either healthy (SCC < 100 × 10³ cells/mL, n = 146) or subclinical mastitis (SCC > 150 × 10³ cells/mL, n = 135) characteristics. Milk was assessed for (i) sLPS/IgG using an “in-house” ELISA, (ii) udder inflammation using LDH activity, and (iii) bacterial presence applying on-farm and standard microbiological laboratory techniques. Mean milk sLPS/IgG absorbances in acute mastitis cows were higher than those detected in healthy and subclinical mastitis cows, with mean differences of 0.35 (95% CI, 0.28 to 0.42) and 0.36 (95% CI, 0.28 to 0.44), respectively. On day 1 of acute mastitis, sLPS/IgG levels in milk containing only Gram-positive bacteria ranged from OD 0.04 to 0.14 (median = 0.1). In contrast, sLPS/IgG levels ranging from OD 0.27 to 1.42 (median = 0.58) and from 0.02 to 1.67 (median = 0.21) were detected in milk containing only Gram-negative bacteria or both Gram-negative and Gram-positive bacteria (i.e., polymicrobial), respectively. Furthermore, differential milk sLPS/IgG absorbance profiles (observed during the testing period days 1–3) were observed in cows with acute mastitis caused by Gram-positive, Gram-negative or polymicrobial infections. Our preliminary findings support the notion that milk sLPS/IgG complexes provide a link between mastitis severity and intramammary Gram-negative infections in dairy cows during early lactation. Full article