Individual quarter dry-off (QDO) is a targeted management strategy used to address persistent IMI that lead to chronic subclinical mastitis, as well as cases of clinical mastitis that are recurrent or unresponsive to treatment. Although the interest in the use of individual QDO as a non-antimicrobial strategy for mastitis control is growing, the impact of this management on subsequent milk production has not been widely explored. Moreover, detailed information on the individual performance of the remaining functional quarters following QDO is scarce. The objective of this observational study was to investigate the effect of lactational QDO following clinical mastitis on short-term milk yield in the remaining individual quarters. This retrospective study was conducted in a commercial dairy farm located in northeast Colorado, equipped with an automatic milking system. The analysis included 114 multiparous cows with one quarter dried off through abrupt cessation of milking following unresponsive clinical mastitis therapy. For comparison, one healthy control cow was matched to each affected cow based on DIM and parity number. Individual quarter milk yield of the remaining functional quarters and from control cows was collected for each milking visit from the on-farm management software and summed as a daily value per quarter for the 30 d following QDO. The herd average DIM at the peak of lactation (68 DIM) was considered to categorize the study cows based on their DIM at QDO into prepeak and postpeak groups. All the analyses were conducted separately for these 2 stage of lactation groups and cows were also categorized based on their dry quarter location (DQL). Least squares means (SE) for daily average milk yield per functional quarter and per cow up to 30 d post QDO were calculated and compared among DQL groups (including matched control cows) using ANOVA for repeated measures analysis, with cow ID as the repeated statement, with compound symmetry selected as the covariance structure. Multivariable models included DQL as explanatory variable of interest and DIM at QDO and calving season as potential covariates. In addition, milk yield curves up to 30 d post-QDO were built for milk yield per DQL using daily LSM calculated by repeated measures analysis. Cow-level milk yield was also compared between DQL groups, including unaffected control cows using <i>t</i>-test for repeated measures analysis. Differences in quarter milk yield were only identified for the prepeak group (≤68 DIM). Milk yield from the left rear and right rear quarters was smaller in control cows than in cows with the right front quarter dried off. When total milk yield per cow was compared within the prepeak group, control cows had greater yield than cows subjected to QDO of the right rear quarter. In the postpeak group, control cows had the highest milk yield compared with the 4 groups of cows with a dry quarter. In conclusion, following QDO, the levels of milk yield compensation in the remaining functional quarters were variable and smaller when QDO occurred post peak (>68 DIM). Nonetheless, in most cases the cow-level milk yield remained lower in 3-quarter cows compared with unaffected controls.

The paper summarizes the clinical experience of Professor JI Laixi in acupuncture treatment for chronic mastitis at lump stage. Professor JI Laixi believes that the essential pathogenesis of this disease is "stagnation". The obstruction in the affected meridians and collaterals due to qi stagnation, qi deficiency, phlegm retention or fire accumulation leads to mastitis. Hence, the treating approach of acupuncture and moxibustion focuses on "relieving stagnation", and 4 principles of treatments are determined, namely regulating qi, tonifying qi, promoting yang circulation and reducing accumulated heat. Clinically, 4 acupuncture techniques are adopted accordingly. Regulating qi and unblocking the circulation in the breast: Hegu (LI4) and Taichong (LR3) are used to relieve masses and stagnation, promote qi movement and improve meridian circulation of the breast. Benefiting qi and regulating qi movement in the breast: Zusanli (ST36), Qihai (CV6) and Neiguan (PC6) are selected to replenish the spleen and stomach, nourish the acquired qi and maintain the free flow of qi in the breast. Promoting yang circulation and resolving masses: the tender points corresponding to Jueyinshu (BL14) and Xinshu (BL15) are stimulated, besides, the patient is required to flex or extend the spine to its maximum position while coordinate with inhalation and exhalation, so as to promote the circulation of heart meridian, propel yang qi movement, warm up and enhance the movements of qi, blood and body fluids to remove stagnation in the breast. Reducing accumulated heat and promoting circulation in the breast: acupuncture is applied to Neiting (ST44), Shangjuxu (ST37), Fenglong (ST40), Neiguan (PC6) and Jianjing (GB21), combined with pricking technique with three-edge needle at Zhongchong (PC9) and Lidui (ST45), and the hooking technique with hook needle at the positive points in the thoracic section of the bladder meridian of foot-taiyang on the back, so as to release the damp-heat stagnation and restore the circulation in the breast.

Mastitis negatively alters milk composition, partly due to oxidative damage to the mammary secretory epithelium. Trace metals are important cofactors involved in oxidative balance and immune function. However, trace metal absorption is affected by its supplement form. The objective of this study was to evaluate milk composition and indicators of oxidative balance in cows supplemented with sulfate or methionine hydroxy analog chelates of Zn, Cu, and Mn at isometal levels during sterile mastitis. A total of 36 mid-lactation multiparous Holstein cows were used. Cows received either (1) sulfate-bound trace metals and intramammary infusions of saline (SULF-CON; n = 9), (2) sulfate-bound trace metals and intramammary infusions of formalin-fixed Staphylococcus aureus (SULF-CHAL; n = 12), or (3) methionine hydroxy analog chelates of Zn, Cu, and Mn and intramammary infusions of formalin-fixed Staph. aureus (MTX-CHAL; n = 12). The study lasted 6 wk, and cows were repeatedly intramammarily infused every 3 d during the final 2 wk to elicit chronic mastitis. Milk yields and components were measured at each milking (twice daily) throughout the challenge period. Blood samples were collected during the final challenge, and mammary tissue biopsies were performed at the end of the study. Milk yields and DMI were similar between treatments. Milk SCS was markedly increased in the MTX-CHAL and SULF-CHAL cows in response to challenge. Milk lactose content decreased in MTX-CHAL and SULF-CHAL cows but tended to be greater in MTX-CHAL cows. The plasma ferric reducing antioxidant power values of SULF-CHAL cows were lower than SULF-CON, whereas the values of MTX-CHAL cows were intermediate. Histological evaluation of mammary tissue structure and markers of lipid peroxidation did not differ between treatments. Results indicate that repeated infusions of formalin-fixed Staph. aureus negatively affected milk composition, and that trace mineral supplement form may better maintain antioxidant status and some markers of epithelial barrier competency during mastitis.

Mastitis negatively affects milk composition, partly due to damage to mammary secretory epithelium. Trace metals are important cofactors for oxidative balance, immune cell function, and maintaining epithelial integrity. The objective of this study was to evaluate blood and milk indices of epithelial barrier integrity in cows supplemented with sulfate or methionine hydroxy analog chelate of Zn, Cu, and Mn at isometal levels during sterile mastitis. A total of 36 mid-lactation multiparous Holstein cows were used. Cows received either (1) sulfate-bound trace metals and intramammary infusions of saline (SULF-CON, n = 9), (2) sulfate-bound trace metals and intramammary infusions of formalin-fixed Staphylococcus aureus (SULF-CHAL, n = 12), or (3) methionine hydroxy analog chelates of Zn, Cu, and Mn and intramammary infusions of formalin-fixed Staph. aureus (MTX-CHAL, n = 12). The study lasted 6 wk, and cows were repeatedly intramammarily infused every 3 d during the final 2 wk to elicit chronic mastitis. Milk composition was assessed at each milking throughout the challenge period, and blood composition was evaluated during the final challenge. The MTX-CHAL and SULF-CHAL cows had increased milk lactate and plasma lactose content. Milk concentrations of Na, S, and Mg increased, whereas Zn, Mn, P, and Fe decreased throughout the challenge period in MTX-CHAL and SULF-CHAL cows. Plasma concentrations of Zn, P, Fe, and Mg decreased during the final challenge in MTX-CHAL and SULF-CHAL cows. Results indicate that repeated infusions of formalin-fixed Staph. aureus did compromise epithelial barrier integrity; however, trace metal supplement form had minimal effect on indices of epithelial function.

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.

Objectives: This study reports the economic impact of common dairy diseases in Jordan. Methods: Data on disease diagnoses and treatments were extracted from electronic farm records of a single, large-scale, intensively managed dairy farm located over a two-year period (2015-2016). During the same period, monthly expenses were obtained and categorized into therapeutic classes, such as antibiotics, anti-inflammatory drugs, hormones, vaccines, disinfectants, and miscellaneous items. For the economic analysis, only variable costs including veterinary fees, medication costs, and estimated discarded milk due to treatment withdrawal periods were considered. Results: Lameness, acute mastitis, endometritis, acute metritis and clinical ketosis were the most significant (p &lt; 0.05) contributors to the disease burden, with prevalence rates of 25%, 20%, 18%, 15%, and 15%, respectively. The most significant (p &lt; 0.05) contributors to the disease economic impact were dystocia with cesarean section (200 JOD, or 282 USD), followed by dystocia without cesarean section (160 JOD, or 225 USD), displaced abomasum (158 JOD, or 222 USD), and acute mastitis (143 JOD, or 201 USD). The average monthly farm expense was 8,088 +- 809 JOD (11,404 +- 1,140 USD), with antibiotics being the most significant (p &lt; 0.05) contributor (2,862 +- 350 JOD, or 4,035 +- 493 USD), followed by vaccines (1,264 +- 150 JOD, or 1,782 +- 211 USD) and disinfectants (1,000 +- 100 JOD, or 1,410 +- 141 USD). Conclusion: These findings highlight the significant economic burden of common diseases on this specific dairy operation and the urgent need for targeted interventions to prevent various diseases affecting dairy cattle.

BackgroundSphingolipids (SL) are key regulators of inflammatory processes, yet their roles in dairy cows remain poorly understood. This study investigated the effects of inflammation (plasma haptoglobin concentration), ketosis, and mastitis on plasma SL profiles in Holstein cows sampled seven days postpartum. From a cohort of 427 cows across 25 farms, 80 animals were classified into four groups: inflammation (n = 20), ketosis (n = 19), mastitis (n = 21), and healthy controls (n = 20). Plasma SL were quantified by targeted HPLC–MS/MS, while cytokines were quantified with a 15-plex bead-based assay. Both univariate and multivariate analyses were applied to assess pathological effects, along with SL ratios and correlations between SL and cytokines.ResultsSystemic inflammation detected through the haptoglobin measure induced the most pronounced alterations in SL metabolism, characterized by elevated dihydrosphingomyelins (DHSM) and lactosylceramides (LacCer), higher C22–24:C16 ratios, and lower unsaturated:saturated ratios in ceramides (Cer) and sphingomyelins (SM). Although total Cer, SM, and the Cer:SM ratio remained unchanged, specific reductions were observed in both Cer and SM in C14, Cer C18:1, SM C16:1, and SM C23:1, whereas SM C25:0 and C26:0 increased. Sphingosine-1-phosphate (So1P) was positively correlated with IL-10 as well as IL-1α and TNFα, while C18–20 Cer correlated positively with multiple pro-inflammatory cytokines and chemokines such as CXCL8 and CCL2. Ketosis induced subtler changes, primarily an increase in plasma DHSM and DHSM:SM ratio (driven by C16:0), an increase in C22–24:C16 DHCer ratio, and a decrease in both LacSo:LacCer and unsaturated:saturated ratios in C23-SM. In this group, So1P correlated positively with CXCL8 and CCL2. Moreover C18–20 Cer and DHCer were positively associated with CXCL8, CCL2, CCL3, and CCL4, which also showed correlations with most LacCer species. Analysis of chronic mastitis cases yielded a clear separation from controls in multivariate analysis but only minimal changes in SL concentrations and ratios, maybe due to the localized nature of the inflammatory response.ConclusionsIn summary, heightened inflammatory response in early post-partum is associated with the strongest systemic effects on SL metabolism, followed by ketosis, while mastitis induced only modest alterations. These findings highlight condition-specific patterns of SL regulation postpartum and suggest potential immunometabolic biomarkers of disease.Supplementary InformationThe online version contains supplementary material available at 10.1186/s40104-025-01325-3.

Chronic Non-bacterial Mastitis (CNBM) is a benign and heterogeneous breast disease whose etiology is not yet fully understood and primarily includes plasma cell mastitis and idiopathic granulomatous mastitis. Its management is challenged by limited therapeutic choices, poor treatment outcomes, and considerable adverse effects. The emergence of molecular targeted therapy has presented new alternatives and hope for the treatment of this condition. The marked dysregulation of critical inflammatory pathways, including NF-κB, JAK-STAT, and MAPK, observed during the onset and progression of CNBM, suggests that molecular agents targeting these signaling cascades may hold therapeutic promise. While preclinical studies in animal models have validated the effectiveness of agents such as pathway inhibitors, clinical evidence is still largely confined to case reports and inferences drawn from other autoimmune diseases. A distinct research gap in the molecular targeted treatment of CNBM persists, owing to the absence of large-scale randomized controlled trials. Advancements in future research will hinge on a multi-pronged approach: utilizing multi-omics methods to establish molecular classifications for precision medicine; designing novel, high-selectivity molecular inhibitors and exploring possibilities for drug repurposing; and studying combined therapeutic regimens to improve efficacy while minimizing toxicity. Through interdisciplinary collaboration and sustained investigation, these initiatives are designed to deliver more effective and safer therapeutic solutions for patients, with the ultimate goal of ameliorating the clinical prognosis of this challenging disease and improving their quality of life.

Acute mastitis remains a major production-limiting disease in dairy buffaloes, particularly during early lactation. The present clinical study evaluated the therapeutic efficacy of a combination of Ceftriaxone and Tazobactam along with multivitamin supplementation in recently calved Murrah buffaloes. Nine clinical cases were subjected to bacteriological examination and antibiogram assay prior to treatment. Ceftriaxone exhibited the highest zone of inhibition (32 mm) against isolated pathogens and was selected for therapy. Animals were treated intramuscularly for five consecutive days. Significant reduction in Somatic Cell Count (SCC) and Total Bacterial Count (TBC) was observed by day 7 and day 16 post-treatment. Complete clinical recovery and improvement in milk quality were recorded, with no recurrence up to 21 days post-therapy. The findings support the use of Ceftriaxone–Tazobactam combination therapy guided by antibiogram testing for effective management of acute mastitis in buffaloes.

ABSTRACTThis study investigated whether the inflammatory component levels in bovine milk measured on the day of mastitis diagnosis were associated with recovery outcomes 7 days later. Milk was collected on Day 0 (the initial day of mastitis diagnosis) and Day 7 from 40 quarters of 32 dairy cows diagnosed with spontaneous acute mastitis. The pH, somatic cell count (SCC), and concentrations of serum amyloid A (SAA), interleukin‐8, lactoferrin (LF), and sodium in milk were measured on Day 0 to evaluate their association with SCC on Day 7. A positive correlation was identified between SCC on Day 7 and SAA, LF, and Na levels on Day 0. For receiver operating characteristic analysis, a log10SCC > 4.8 (70,000 cells/mL) on Day 7 was considered indicative of mastitis. The cutoff values for SAA and LF on Day 0 were 21.6 and 81 μg/mL, respectively, with sensitivities of 79.3% and 86.2%, specificities of 77.8% and 66.7%, and areas under the curve of 0.784 and 0.762, respectively. These results indicate that measuring SAA and LF concentrations in milk at the initial mastitis diagnosis may predict intramammary conditions 7 days later, potentially aiding in understanding the pathology of mastitis and informing treatment strategies.

Introduction Recently, neutralizing autoantibodies against different cytokines have been found to explain susceptibility to infections. Because the clinical pictures mimic innate immune disorders (IIDs), they have been defined as phenocopies of IIDs. Autoantibodies against GM-CSF have recently been associated with disseminated cryptococcosis and alveolar proteinosis. Clinical Case The patient was a 17-year-old female with no family or personal history of disease. She presented with clinical symptoms for one year, characterized by fainting episodes, headaches, and vomiting. She underwent multiple medical evaluations, but no diagnostic conclusions were reached. Over time, the headache intensity increased and was associated with fever, asthenia, refusal of food, and subcutaneous lesions on the abdomen and chest that appeared to be lipomas. Ultimately, her condition was associated with mastitis, a solid nodule, and inflamed lymph nodes in the axillary regions. The biopsy revealed chronic granulomatous mastitis with Langhans giant cells and structures consistent with Cryptococcus spp. The cytochemical study of the cerebrospinal fluid showed hyperproteinorrhea and hypoglycorrhachia. The brain MRI revealed a lesion in the right basal ganglia related to cryptococcosis. Disseminated cryptococcosis was confirmed, and she completed six weeks of amphotericin B. It was suspended due to kidney damage and changed to fluconazole, which remains on to date. The immunological approach included a complete blood count, immunoglobulins, and lymphocyte subpopulations with normal values for her age. The autoimmunity approach and HIV test were negative. Antibodies against cytokines were requested, with identification of anti–GM-CSF autoantibodies. Discussion In all patients with disseminated cryptococcosis, we must look for immunological abnormalities. If no monogenic defects are found, we must look for autoantibodies against GM-CSF. In Mexico, we have implemented this search in the immunodeficiency laboratory. Patients may develop alveolar proteinosis, so they should be followed up by a pneumologist. Autoantibodies are compatible with autoimmunity, so treatment is immunosuppressive and immunomodulatory.

Lycopene exerts a treatment effect on bovine mastitis induced by Escherichia coli infection via the MAPK pathway using network pharmacology, a mice model and molecular docking.

Background: Acute or chronic mastitis in non-lactating female is rising now a days.There is a spectrum of etiological factors ranging from infection to autoimmune disorders.The distinction among different forms of mastitis is of great importance since the management of these different entities varies.This study aimed to explore the clinical presentation and etiological factors of various mastitis in non-lactating female during their reproductive period. Methods:A cross-sectional descriptive study was carried out on 100 female patients of reproductive period presented with the complains of different varieties of mastitis non related to lactation between 01-04-2018 to 30-04-2019 in Ad-Din Women's Medical College & Hospital.Patients were evaluated by history, clinical examination, imaging, FNACor tru cut biopsy, histopathology & pus culture. Results:We found out of 100 patients, 58 cases were breast abscess (58%), 19 cases were granulomatous mastitis (19%), 11 cases were ANDI (fibrocystic disease) (11%), 8 cases were Idiopathic granulomatous mastitis (8%) & 4 cases were periductal mastitis (4%).The age of the patients was in between 18 to 50 years & the mean age was in between 31.777.99 year.Staphylococcus aureus was most commonly isolated organism (61.97%).Rest of the patients culture showed S. epidermidis in 8.45% patients & Klebsiella in 2.81% patients.No growth was found in 26.76% cases.conclusion: Non-lactational mastitis causes considerable morbidity & psychological distress in females.The purpose of this study was to analyze spectrum of mastitis in non-lactating females thus help to provide prompt & appropriate management to prevent complications.

Staphylococcus aureus is the leading cause of acute mastitis during lactation. This species has a remarkable ability to form biofilms and to develop antibiotic resistance, which hampers the effectiveness of current therapeutic approaches. This study aims to evaluate the therapeutic potential of three lactic acid bacteria (LAB) strains (Limosilactobacillus fermentum I7, Limosilactobacillus reuteri 7SNG3-30 and Ligilactobacillus salivarius 22SNG3-30) to interfere with biofilms formed by two S. aureus strains isolated from milk of women with acute mastitis. Both live LAB cells and their respective cell-free supernatants were able to disrupt the S. aureus biofilm structure and significantly reduce its cellular viability. However, the effectiveness of these treatments was dependent on the S. aureus strain, the LAB strain, and the type of LAB preparation (active culture or cell-free supernatant) involved in each interaction. Overall, our results suggest that the tested LAB strains have the potential to be used either as probiotics or postbiotics complementing the current therapies against acute mastitis and other staphylococcal infections.

Lactation mastitis is a common postpartum disease. Flucloxacillin is frequently used for treatment, but its efficacy is limited in some cases, potentially causing adverse outcomes such as premature cessation of breastfeeding. This study aimed to identify risk factors for poor treatment outcomes in lactational mastitis managed with flucloxacillin and provide evidence for precise clinical management. A retrospective cohort study was conducted from January to December 2022 at the Chongqing Health Center for Women and Children in China. It included 133 hospitalized patients with acute lactational mastitis treated with flucloxacillin, divided into effective treatment (n = 93) and treatment failure (n = 40) groups. Effective treatment was defined as clinical improvement characterized by a reduction in breast pain and swelling, along with normalization of body temperature within 48 hours of initiating flucloxacillin therapy. Treatment failure was defined as the absence of clinical improvement or worsening of symptoms, necessitating a change in the antimicrobial regimen. Data on patient demographic and clinical characteristics, laboratory findings, and treatment outcomes were collected and analyzed. Among 133 patients, 93 (69.9%) were effectively treated with flucloxacillin, while 40 (30.1%) experienced treatment failure. Binary logistic regression identified onset time ≥3 days (Adjusted OR 2.76, 95% CI 1.22, 6.28) and inflammation located in the nipple/areola area (Adjusted OR 3.28, 95% CI 1.27, 8.49) as independent risk factors for treatment failure. The treatment failure group had longer median fever duration (3 days vs. 1 day, p < 0.001), longer median hospital stay (7 days vs. 4 days, p < 0.001), higher median hospitalization costs (3868 yuan [US$553] vs. 2000 yuan [US$286], p < 0.001), and higher lactation suppression rates (20% vs. 2.2%, p = 0.001). Staphylococcus aureus was the predominant isolate, with a higher isolation rate in the treatment effective group (88% vs. 50%, OR 7.0, 95% CI 1.78, 27.42). The present study identified that treatment failure of flucloxacillin for acute lactational mastitis was significantly associated with disease onset ≥3 days, inflammation in the nipple/areola area, and non-Staphylococcus aureus infections. Early identification and intervention are essential for better outcomes. Developing guidelines that emphasize early intervention may contribute to reducing fever duration, patient discomfort, breastfeeding cessation rates, and healthcare burden.

Rare breast tumors

Background: Caprine arthritis encephalitis (CAE) is a major viral disease of goats, caused by small ruminant lentivirus (SRLV), associated with chronic arthritis, mastitis, pneumonia, and encephalitis, leading to economic losses and reduced animal welfare. This study aimed to estimate the true prevalence of CAE in Hungarian goat herds, based on nationwide sampling and statistical modeling. Methods: Blood samples from 1218 goats in 53 herds were tested using ELISA, and true prevalence was estimated by Bayesian analysis. Results: The mean herd true prevalence (HTP) was 29.1% (95% CrI: 20.8–38.5%), while within the infected herds, the conditional within herd prevalence (CWHP) reached 58% ± 27.1%. Medium- and large-sized herds (>50 animals) showed the highest mean HTP (77.8% and 74.9%, respectively). No significant regional differences were observed, indicating that CAE is uniformly distributed across the country. Conclusions: Our findings place Hungary among moderately to highly affected European countries and highlight the need for a nationwide control strategy integrating routine serological surveillance, biosecurity improvements, farmer education, and long-term tools such as selective breeding.

Mastitis is a common pathological condition in dairy cattle. If not promptly and effectively treated, it can progress to chronic subclinical mastitis, which is characterized by fibrosis of mammary gland tissue and results in a marked decline in both milk yield and quality. During chronic inflammation, transforming growth factor-β1 (TGF-β1) is often overexpressed, driving epithelial-mesenchymal transition (EMT) and subsequent tissue fibrosis. However, the molecular mechanisms by which TGF-β1 induces EMT in bovine mammary epithelial cells (BMECs) remain poorly understood. The compound 6-bromo-indirubin 3'-oxime (6BIO) is known for its anti-inflammatory, antibacterial, and anti-proliferative activities, but its role in EMT regulation has not been fully defined. Using primary BMEC cultures, this study examined the regulatory function of TGF-β1 in EMT induction and evaluated the modulatory effects of 6BIO on cell morphology, inflammatory responses, and EMT progression. Results showed that TGF-β1 triggered inflammation in BMECs, promoted their phenotypic transition to an interstitial state, induced EMT, and activated the TGF-β/Smad signaling pathway. In contrast, 6BIO inhibited this phenotypic transition, suppressed TGF-β1-induced EMT, and blocked TGF-β/Smad activation. Additionally, 6BIO directly interacted with Smad3 proteins. Overall, these findings indicate that aberrant TGF-β1 upregulation promotes EMT in BMECs, whereas 6BIO counteracts this process through interaction with Smad3. This study provides new insights into strategies for preventing and managing chronic subclinical mastitis and establishes a basis for developing anti-EMT therapeutic agents.

Acute mastitis is a common infection during lactation, primarily caused by Staphylococcus aureus, a bacterium known for its ability to form biofilms within mammary ducts and develop antibiotic resistance. This study aimed to genomically characterize S. aureus strains isolated from women with acute mastitis and healthy asymptomatic women to better understand how S. aureus strains transition from harmless components of the human milk microbiota to pathogenic agents responsible for mastitis. Whole-genome sequencing was performed on nine S. aureus strains-six from women with mastitis and three from healthy women-followed by in silico analyses of core and accessory genes, resistome, virulome, mobilome, and secondary metabolite synthesis to identify genes related to virulence, antibiotic resistance, biofilm formation, and mobile genetic elements such as plasmids, bacteriophages, and pathogenicity islands. Antimicrobial resistance profiles were evaluated using Sensititre EUST2 plates, antimicrobial activity by an agar diffusion method, biofilm formation in 96-well plates, and siderophore production with the Chrome Azurol S assay. Results showed that complete bacteriophage genomes were only present in S. aureus strains isolated from mastitis cases. Some virulence genes, including fnbB and cna, were absent in strains from healthy women. Both types of S. aureus strains exhibited biofilm formation capacity, with mastitis-associated strains SA4 and SA5 being the highest biofilm producers. Similarly, although all strains secreted siderophores, SA4 and SA55 exhibited the strongest siderophore production, indicating a link between this trait and virulence in mastitis-associated strains. The analysis of key genomic features, including virulence factors, resistance genes, and biofilm-forming capabilities, revealed some mechanisms by which S. aureus contributes to the pathogenesis of mastitis.IMPORTANCEAcute mastitis is a widespread infection in lactating women, and its main cause, Staphylococcus aureus, has developed resistance to antibiotics, making treatment challenging. The ability of this bacterium to form biofilms complicates its eradication from the mammary glands. Understanding the genomic and phenotypic characteristics of S. aureus strains associated with mastitis, compared to those isolated from asymtomatic women, is critical for developing better treatment strategies. This study provides new insights into the genetic features, such as virulence factors, antibiotic resistance profiles, and presence of bacteriophages, that make S. aureus strains pathogenic in mastitis. It also highlights the potential of biofilm formation and siderophore production as key factors in mastitis progression. These findings could guide the development of novel therapeutic approaches, such as targeted therapies or probiotics, which can more effectively treat mastitis and reduce reliance on antibiotics, ultimately improving maternal and infant health.

Bovine mastitis is one of the most prevalent and economically significant diseases affecting dairy cows worldwide, with Escherichia coli (E. coli) recognized as one of the principal pathogens causing acute mastitis. The innate immune system plays a crucial role in the defense of the bovine mammary gland, serving as the first line of defense against pathogen invasion. This study elucidated the pathological mechanisms and immune response-related molecular regulatory networks involved in E. coli-induced bovine mastitis. Histopathological and apoptosis analyses of mammary tissues were performed using hematoxylin-eosin (HE) staining and TUNEL staining, respectively, while RNA sequencing (RNA-seq) was conducted to identify differentially expressed genes (DEGs) and their associated signaling pathways. HE staining revealed typical inflammatory lesions in the mammary glands of mastitis cows. TUNEL staining further confirmed that the level of apoptosis in the mastitis group was significantly higher than in the healthy control group (p < 0.0001). RNA-seq analysis identified 2717 DEGs, with 2238 upregulated and 479 downregulated genes. The top 20 significantly upregulated genes (e.g., S100A12, IL1RN, IL1R2, CXCL8, SAA3, S100A8, S100A9, TREML2, TREM1, M-SAA3.2, PTX3, MMP9) were predominantly involved in inflammatory immune regulation, acute phase responses (e.g., HP, SAA3), and cellular signal transduction (e.g., PLEK, LPAR3). Gene Ontology (GO) enrichment analysis revealed that these DEGs were mainly associated with biological processes, such as signal transduction, immune response, inflammatory response, and transcriptional regulation. Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis indicated that these DEGs were significantly enriched in key inflammatory and immune regulatory pathways, including the TNF signaling pathway, C-type lectin receptor signaling pathway, Chemokine signaling pathway, NOD-like receptor signaling pathway, NF-κ B signaling pathway, and IL-17 signaling pathway, suggesting that these pathways play central roles in the mammary immune defense against E. coli infection. In conclusion, this study demonstrated at the histopathological, cellular apoptosis, and transcriptomic levels that E. coli infection induces mammary tissue damage and apoptosis by activating immune and inflammation-related genes (S100A12, IL1RN, IL1R2, CXCL8, SAA3, S100A8, S100A9, TREML2, TREM1, M-SAA3.2, PTX3, MMP9) and key signaling pathways (TNF signaling pathway, C-type lectin receptor signaling pathway, Chemokine signaling pathway, NOD-like receptor signaling pathway, NF-κ B signaling pathway, IL-17 signaling pathway). The findings of this study provide a theoretical basis for probing into the pathogenesis of bovine mastitis and the development of targeted interventions.