Enterogenic Stenotrophomonas maltophilia migrates to the mammary gland to induce mastitis by activating the calcium-ROS-AMPK-mTOR-autophagy pathway

Background Mastitis is an inflammatory disease of the mammary gland that has serious economic impacts on the dairy industry and endangers food safety.Our previous study found that the body has a gut/rumen-mammary gland axis and that disturbance of the gut/rumen microbiota could result in‘gastroenterogenic mastitis'.However,the mechanism has not been fully clarified.Recently,we found that long-term feeding of a high-concentrate diet induced mastitis in dairy cows,and the abundance of Stenotrophomonas maltophilia(S.maltophilia)was significantly increased in both the rumen and milk microbiota.Accordingly,we hypothesized that‘gastroenterogenic mastitis'can be induced by the migration of endogenous gut bacteria to the mammary gland.Therefore,this study investigated the mechanism by which enterogenic S.maltophilia induces mastitis.Results First,S.maltophilia was labelled with superfolder GFP and administered to mice via gavage.The results showed that treatment with S.maltophilia promoted the occurrence of mastitis and increased the permeability of the blood-milk barrier,leading to intestinal inflammation and intestinal leakage.Furthermore,tracking of ingested S.maltophilia revealed that S.maltophilia could migrate from the gut to the mammary gland and induce mastitis.Subsequently,mammary gland transcriptome analysis showed that the calcium and AMPK signalling pathways were significantly upregulated in mice treated with S.maltophilia.Then,using mouse mammary epithelial cells(MMECs),we verified that S.maltophilia induces mastitis through activation of the calcium-ROS-AMPK-mTOR-autophagy pathway.Conclusions In conclusion,the results showed that enterogenic S.maltophilia could migrate from the gut to the mammary gland via the gut-mammary axis and activate the calcium-ROS-AMPK-mTOR-autophagy pathway to induce mastitis.Targeting the gut-mammary gland axis may also be an effective method to treat mastitis.

Granulomatous mastitis in a 50-year-old male:A case report and review of literature

BACKGROUND Granulomatous mastitis(GM)an inflammatory disease of the breast that usually affects women of childbearing age,occurs very rarely in males.CASE SUMMARY We present a case study of a 50-year-old male patient with GM.The patient developed a breast lump following the cleaning of a previously embedded dirtfilled nipple.While an initial improvement was noted with antibiotic therapy,a recurrence occurred a year later,showing resistance to the previously effective antibiotics.Subsequently,the lesion was excised.The histopathological examination confirmed the diagnosis of GM.CONCLUSION GM should be considered a possible diagnosis of male breast masses.

Methylome and transcriptome data integration reveals potential roles of DNA methylation and candidate biomarkers of cow Streptococcus uberis subclinical mastitis

Background:Mastitis caused by different pathogens including Streptococcus uberis(S.uberis)is responsible for huge economic losses to the dairy industry.In order to investigate the potential genetic and epigenetic regulatory mecha‑nisms of subclinical mastitis due to S.uberis,the DNA methylome(whole genome DNA methylation sequencing)and transcriptome(RNA sequencing)of milk somatic cells from cows with naturally occurring S.uberis subclinical mastitis and healthy control cows(n=3/group)were studied.Results:Globally,the DNA methylation levels of CpG sites were low in the promoters and first exons but high in inner exons and introns.The DNA methylation levels at the promoter,first exon and first intron regions were nega‑tively correlated with the expression level of genes at a whole‑genome‑wide scale.In general,DNA methylation level was lower in S.uberis‑positive group(SUG)than in the control group(CTG).A total of 174,342 differentially methylated cytosines(DMCs)(FDR<0.05)were identified between SUG and CTG,including 132,237,7412 and 34,693 DMCs in the context of CpG,CHG and CHH(H=A or T or C),respectively.Besides,101,612 methylation haplotype blocks(MHBs)were identified,including 451 MHBs that were significantly different(dMHB)between the two groups.A total of 2130 differentially expressed(DE)genes(1378 with up‑regulated and 752 with down‑regulated expression)were found in SUG.Integration of methylome and transcriptome data with MethGET program revealed 1623 genes with signifi‑cant changes in their methylation levels and/or gene expression changes(MetGDE genes,MethGET P‑value<0.001).Functional enrichment of genes harboring≥15 DMCs,DE genes and MetGDE genes suggest significant involvement of DNA methylation changes in the regulation of the host immune response to S.uberis infection,especially cytokine activities.Furthermore,discriminant correlation analysis with DIABLO method identified 26 candidate biomarkers,including 6 DE genes,15 CpG‑DMCs and 5 dMHBs that discriminated between SUG and CTG.Conclusion:The integration of methylome and transcriptome of milk somatic cells suggests the possible involve‑ment of DNA methylation changes in the regulation of the host immune response to subclinical mastitis due to S.uberis.The presented genetic and epigenetic biomarkers could contribute to the design of management strategies of subclinical mastitis and breeding for mastitis resistance.

Heterogeneity of Outcomes Reporting in Trials Evaluating Traditional Chinese Medicine Breast Massage for Stasis Acute Mastitis: A Methodological Review

Objective To systematically analyze the current status of outcomes reporting in clinical trials on treating stasis acute mastitis with Traditional Chinese Medicine breast massage.Methods We searched CNKI,Wanfang,VIP,Sino Med,Pub Med,Web of Science,Embase,Cochrane library,JBI,CINAHL,Psyc INFO,Clinical Trials Registry Platform portal,Clinical Trials Registry,Australian New Zealand Clinical Trials Registry,Center Watch Registry from inception to May 15,2022 to find randomized controlled trials,non-randomized controlled trials,case series and cohort studies which reported the outcomes of stasis acute mastitis managed with Traditional Chinese Medicine breast massage,with search terms of mastitis,acute mastitis,lactation mastitis,puerperal mastitis,breast problem,breast engorgement,milk stasis,blocked ducked,breast pain,breast massage,and acupoint massage.Outcomes and the measurement schemes(measurement methods,timing of assessing outcome,frequency of assessing outcome,measurers)were extracted from the included studies.We used the Management of Otitis Media with Effusion in Children with Cleft Palate(MOMENT)to assess the quality of each study,then categorized outcomes derived from the included studies into different domains according to the Outcome Measures in Rheumatology Arthritis Clinic Trials(OMERACT)Filter 2.1 framework.Results We identified 85 clinical trials,in which 54 different outcomes were reported.A total of 81.2%(69/85)of studies were assessed as medium quality with a mean score of 2.6,and 18.8%(16/85)as low quality with a mean score of 0.9.These outcomes were organized in three core areas.Lump size(89.4%,76/85)was the most frequently reported outcome,followed by breast pain(69.4%,59/85)and milk excretion(68.2%,58/85).Five methods were used to assess lump size and four methods to assess breast pain.Conclusions The outcomes reported in clinical trials regarding stasis acute mastitis treated by Traditional Chinese Medicine breast massage are heterogeneous.Developing a core outcome set to achieve consistent standards for reporting outcomes and modalities for validation of the outcomes is clearly warranted.

Candidate genes for mastitis resistance in dairy cattle:a data integration approach

Background Inflammation of the mammary tissue(mastitis)is one of the most detrimental health conditions in dairy ruminants and is considered the most economically important infectious disease of the dairy sector.Improving mastitis resistance is becoming an important goal in dairy ruminant breeding programmes.However,mastitis resistance is a complex trait and identification of mastitis-associated alleles in livestock is difficult.Currently,the only applicable approach to identify candidate loci for complex traits in large farm animals is to combine different information that supports the functionality of the identified genomic regions with respect to a complex trait.Methods To identify the most promising candidate loci for mastitis resistance we integrated heterogeneous data from multiple sources and compiled the information into a comprehensive database of mastitis-associated candidate loci.Mastitis-associated candidate genes reported in association,expression,and mouse model studies were collected by searching the relevant literature and databases.The collected data were integrated into a single database,screened for overlaps,and used for gene set enrichment analysis.Results The database contains candidate genes from association and expression studies and relevant transgenic mouse models.The 2448 collected candidate loci are evenly distributed across bovine chromosomes.Data integration and analysis revealed overlaps between different studies and/or with mastitis-associated QTL,revealing promising candidate genes for mastitis resistance.Conclusion Mastitis resistance is a complex trait influenced by numerous alleles.Based on the number of independent studies,we were able to prioritise candidate genes and propose a list of the 22 most promising.To our knowledge this is the most comprehensive database of mastitis associated candidate genes and could be helpful in selecting genes for functional validation studies.

Transcriptome‑wide mapping of milk somatic cells upon subclinical mastitis infection in dairy cattle

Background Subclinical intramammary infection(IMI)represents a significant problem in maintaining dairy cows’health.Disease severity and extent depend on the interaction between the causative agent,environment,and host.To investigate the molecular mechanisms behind the host immune response,we used RNA-Seq for the milk somatic cells(SC)transcriptome profiling in healthy cows(n=9),and cows naturally affected by subclinical IMI from Proto-theca spp.(n=11)and Streptococcus agalactiae(S.agalactiae;n=11).Data Integration Analysis for Biomarker discov-ery using Latent Components(DIABLO)was used to integrate transcriptomic data and host phenotypic traits related to milk composition,SC composition,and udder health to identify hub variables for subclinical IMI detection.Results A total of 1,682 and 2,427 differentially expressed genes(DEGs)were identified when comparing Prototheca spp.and S.agalactiae to healthy animals,respectively.Pathogen-specific pathway analyses evidenced that Proto-theca’s infection upregulated antigen processing and lymphocyte proliferation pathways while S.agalactiae induced a reduction of energy-related pathways like the tricarboxylic acid cycle,and carbohydrate and lipid metabolism.The integrative analysis of commonly shared DEGs between the two pathogens(n=681)referred to the core-mastitis response genes,and phenotypic data evidenced a strong covariation between those genes and the flow cytometry immune cells(r2=0.72),followed by the udder health(r2=0.64)and milk quality parameters(r2=0.64).Variables with r≥0.90 were used to build a network in which the top 20 hub variables were identified with the Cytoscape cyto-hubba plug-in.The genes in common between DIABLO and cytohubba(n=10)were submitted to a ROC analysis which showed they had excellent predictive performances in terms of discriminating healthy and mastitis-affected animals(sensitivity>0.89,specificity>0.81,accuracy>0.87,and precision>0.69).Among these genes,CIITA could play a key role in regulating the animals’response to subclinical IMI.Conclusions Despite some differences in the enriched pathways,the two mastitis-causing pathogens seemed to induce a shared host immune-transcriptomic response.The hub variables identified with the integrative approach might be included in screening and diagnostic tools for subclinical IMI detection.

Clinical efficacy of external treatment with Zi Cao Yang He Tang combined with hormone therapy for granulomatous mastitis and its effect on cellular pyroptosis proteins

Objective:To observe the clinical effect of external application of Zi Cao Yang He Tang combined with dexamethasone ointment in the treatment of granulomatous lobular mastitis in the mass stage and the effect on cellular pyroptosis-related proteins.Methods:Forty patients with granulomatous lobular mastitis were randomly divided into 20 cases in the observation group and 20 cases in the control group.Both groups were given dexamethasone ointment for treatment,and the observation group was treated with external application of Zi Cao Yang He Tang on the basis of the control group.Breast lump vol-ume changes,breast pain and pus symptoms,pathology and histology and immunohistochemistry,cellular death-related indicators expression were compared between the two groups.Results:After treatment,the difference in mass volume of the observation group was significantly greater than that of the control group(16.7±21.5 in the control group and 32.9±27.4 in the experimental group)and the difference between the two groups was statistically significant(P<0.05);the objective remission rate of the patients in the observation group(ORR=85%)was significantly higher than that of the treatment group(ORR=50%,P<0.05).In terms of pus formation rate and pain level,the breast pain score of the observation group was significantly lower than that of the control group after two weeks or three weeks of treatment(P<0.05);the pus formation rate of the observation group was 30%,and the pus formation rate of the control group was 20%,and the difference was not statistically significant(P>0.05).The immunohistochemical expression levels of caspase-1 and GSDMD in the observation group were significantly lower than those in the control group(P<0.05);and in terms of the expression of cellular pyroptosis-related indexes,the expression of GSDMD,cas-pase-1,and NLRP3 in the observation group was significantly lower compared with that in the control group(P<0.05).Conclusion:The efficacy of external application of Zi Cao Yang He Tang combined with dexamethasone ointment in treating granulomatous lobular mastitis in the mass stage is remarkable,which can effectively reduce breast pain,reduce the size of the mass,de-crease the expression of cellular pyroptosisrelated indexes,and have a good anti-inflammatory effect.

Therapeutic Effect of Daphnetin on Mastitis Induced by Staphylococcus aureus in Mice

[Objectives]To observe the effects of daphnetin on mastitis induced by Staphylococcus aureus in mice.[Methods]18 postpartum ICR female mice were used to establish mastitis animal model,and were randomly divided into three groups(A,B,and C)with 6 mice in each group.Group A:blank control group;group B:S.Aureus model group;group C:S.Aureus model+daphnetin group.The experimental groups were injected 1 mL of 1.0×104 CFU/100μL of S.aureus of along the nipple catheter.The suspension was placed in the 3 rd and 4 th pairs of mammary glands,and the control group was injected with the same dose of normal saline.On the second day after infection,the rats in group A,B and C were given drugs by gavage,while the rats in group A and B were given normal saline and the rats in group C were given daphnetin once a day for 6 consecutive days.Blood samples were collected from living eyeballs,and blood cells were analyzed by automatic flow cytometer after anticoagulation.[Results]The NLR and Systemie Immune Inflammati-on Index(SII)in the blood of mastitis mice induced by S.aureus were significantly higher than those in the control group(P<0.01),suggesting that neutrophil to lymphocyte ratio(NLR)and SII can be used as diagnostic indicators of mastitis,and the levels of NLR and SII decreased significantly after daphnetin intervention.[Conclusions]NLR and SII showed high levels in mastitis mice,which are valuable for the diagnosis of mastitis and the evaluation of its prognosis.After the intervention of daphnetin,both of them decreased significantly,indicating that daphnetin has a good prognosis trend in mastitis mice induced by S.aureus.

Exploring the pathogenesis of idiopathic granulomatous mastitis based on the prolactin theory

Prolactin(PRL),as a cytokine,plays a variety of multidirectional roles in immunity and inflammation.An autoimmune condition known as idiopathic granulomatous mastitis(IGM)involves many inflammatory cells and cytokines.PRL links neuroendocrine and immune systems and plays an active role in the inflammatory and immune processes.Based on the theory of PRL and combined with the latest research review at home and abroad,this paper aims to review the mechanism of action of PRL on IGM from the aspects of various inflammatory cells,cytokines and pathways,revealing the influence of PRL theory on IGM and the mechanism of action,so as to effectively control inflammation in the long-term.

Management of granulomatous lobular mastitis: an international multidisciplinary consensus(2021 edition)

Granulomatous lobular mastitis(GLM) is a rare and chronic benign inflammatory disease of the breast. Difficulties exist in the management of GLM for many front-line surgeons and medical specialists who care for patients with inflammatory disorders of the breast. This consensus is summarized to establish evidence-based recommendations for the management of GLM. Literature was reviewed using PubMed from January 1, 1971 to July 31, 2020. Sixty-six international experienced multidisciplinary experts from 11 countries or regions were invited to review the evidence.Levels of evidence were determined using the American College of Physicians grading system, and recommendations were discussed until consensus. Experts discussed and concluded 30 recommendations on historical definitions,etiology and predisposing factors, diagnosis criteria, treatment, clinical stages, relapse and recurrence of GLM. GLM was recommended as a widely accepted definition. In addition, this consensus introduced a new clinical stages and management algorithm for GLM to provide individual treatment strategies. In conclusion, diagnosis of GLM depends on a combination of history, clinical manifestations, imaging examinations, laboratory examinations and pathology.The approach to treatment of GLM should be applied according to the different clinical stage of GLM. This evidencebased consensus would be valuable to assist front-line surgeons and medical specialists in the optimal management of GLM.

Granulomatous Mastitis, Radio-Histological Correlation

Granulomatous mastitis, also called granulomatous lobular mastitis, is a benign pathology of the mammary gland characterized by chronic inflammation of the gland tissue, which becomes erythematous, manifesting itself as a palpable and painful mass on physical examination, sometimes it can be accompanied by abscesses. The following case is a 45-year-old Guatemalan female, with a history of increased volume in the left breast, compromising the upper quadrants. A mammogram was performed, followed by breast ultrasound;the diagnosis was confirmed by histology. The treatment offered was surgery with image controls every six months.

Aetiology of idiopathic granulomatous mastitis

Idiopathic granulomatous mastitis is a rare chronic inflammatory lesion of the breast that can clinically and radiographically mimic breast carcinoma. The most common clinical presentation is an unilateral, discrete breast mass, nipple retraction and even a sinus formation often associated with an inflammation of the overlying skin. The etiology of idiopathic granulomatous mastitis is still obscure. Its treatment remains controversial. The cause may be the autoimmune process, infection, a chemical reaction associated with oral contraceptive pills, or even lactation. Various factors, including hormonal imbalance, autoimmunity, unknown microbiological agents, smoking and α 1-antitrypsin deficiency have been suggested to play a role in disease aetiology. In this review, causing factors in the aetiology of idiopathic granulomatous mastitis are reviewed in detail.

Granulomatous lobular mastitis secondary to Mycobacterium fortuitum

Granulomatous lobular mastitis is a rare inflammatory disease of the breast of unknown etiology. Most present as breast masses in women of child-bearing age. A 29-year-old female presented with a swollen, firm and tender right breast, initially misdiagnosed as mastitis. Core needle biopsy revealed findings consistent with granulomatous lobular mastitis, and cultures were all negative for an infectious etiology. She was started on steroid therapy to which she initially responded well. A few weeks later she deteriorated and was found to have multiple breast abscesses. She underwent operative drainage and cultures grew Mycobacterium fortuitum. Granulomatous lobular mastitis is a rare inflammatory disease of the breast. The definitive diagnose entails a biopsy. Other causes of chronic or granulomatous mastitis should be ruled out, including atypical or rare bacteria such as Mycobacterium fortuitum. This is the first reported case of granulomatous mastitis secondary to Mycobacterium fortuitum. With pathologic confirmation of granulomatous mastitis, an infectious etiology must be ruled out. Atypical bacteria such as Mycobacterium fortuitum may not readily grow on cultures, as with our case. Medical management is appropriate, with surgical excision reserved for refractory cases or for drainage of abscesses.

Genetic characterization of antimicrobial resistance in Staphylococcus aureus isolated from bovine mastitis cases in Northwest China

Staphylococcus aureus is the most common etiological pathogen of bovine mastitis. The resistant strains make the disease difficult to cure. The aim of this study was to characterize the genetic nature of the antimicrobial resistance in S. aureus cultured from bovine mastitis in Northwest China in 2014. A total of 44 S. aureus were isolated for antimicrobial resistance and resistance-related genes. Antimicrobial resistance was determined by disc diffusion and the corresponding resistance genes were detected by PCR. Phenotype indicated that S. aureus isolates were resistant to penicillin （84.09%）, erythromycin （20.45%）, tetracycline （15.91%）, gentamicin （9.09%）, tobramycin （6.82%）, kanamycin （6.82%） and methicillin （2.27%）. 9.09% of the S. aureus isolates were classified as multidrug resistant. In addition, genotypes showed that the isolates were resistant to rifampicin （100%, rpoB）, penicillin （95.45%, blaZ）, tetracycline （22.73%, tetK, tetM, alone or in combination）, erythromycin （22.73%, ermB or ermC）, gentamicin/tobramycin/kanamycin （2.27%, aacA-aphD）, methicillin （2.27%, mecA） and vancomycin （2.27%, vanA）. Resistance to tetracycline was attributed to the genes tetK and tetM （r=0.558, P〈0.001）. This study noted high-level geno- and phenotypic antimicrobial resistance in S. aureus isolates from bovine mastitis cases in Northwest China.

Rumen microbiome structure and metabolites activity in dairy cows with clinical and subclinical mastitis

Background:Due to the high prevalence and complex etiology,bovine mastitis(BM)is one of the most important diseases to compromise dairy cow health and milk quality.The shift in milk compositions has been widely investigated during mastitis,but recent studies suggested that gastrointestinal microorganism also has a crucial effect on the inflammation of other peripheral tissues and organs,including the mammary gland.However,research focused on the variation of rumen inner-environment during mastitis is still limited.Therefore,the ruminal microbial profiles,metabolites,and milk compositions in cows with different udder health conditions were compared in the present study.Furthermore,the correlations between udder health status and ruminal conditions were investigated.Based on the somatic cell counts(SCC),California mastitis test(CMT)parameters and clinical symptoms of mastitis,60 lactating Holstein dairy cows with similar body conditions(excepted for the udder health condition)were randomly divided into 3 groups(n=20 per group)including the healthy(H)group,the subclinical mastitis(SM)group and the clinical mastitis(CM)group.Lactation performance and rumen fermentation parameters were recorded.And rumen microbiota and metabolites were also analyzed via 16S rRNA amplicon sequencing and untargeted metabolomics,respectively.Results:As the degree of mastitis increased,rumen lactic acid(LA)(P<0.01),acetate,propionate,butyrate,valerate(P<0.001),and total volatile fatty acids(TVFAs)(P<0.01)concentrations were significantly decreased.In the rumen of CM cows,the significantly increased bacteria related to intestinal and oral inflammation,such as Lachnospiraceae(FDR-adjusted P=0.039),Moraxella(FDR-adjusted P=0.011)and Neisseriaceae(FDR-adjusted P=0.036),etc.,were accompanied by a significant increase in 12-oxo-20-dihydroxy-leukotriene B4(FDR-adjusted P=5.97×10^(−9))and 10beta-hydroxy-6beta-isobutyrylfuranoeremophilane(FDR-adjusted P=3.88×10^(−10)).Meanwhile,in the rumen of SM cows,the Ruminiclostridium_9(FDR-adjusted P=0.042)and Enterorhabdus(FDR-adjusted P=0.043)were increased along with increasing methenamine(FDR-adjusted P=6.95×10^(−6)),5-hydroxymethyl-2-furancarboxaldehyde(5-HMF)(FDR-adjusted P=2.02×10^(−6))and 6-methoxymellein(FDR-adjusted P=2.57×10^(−5)).The short-chain fatty acids(SCFAs)-producing bacteria and probiotics in rumen,including Prevoterotoella_1(FDRadjusted P=0.045)and Bifidobacterium(FDR-adjusted P=0.035),etc.,were significantly reduced,with decreasing 2-phenylbutyric acid(2-PBA)(FDR-adjusted P=4.37×10^(−6)).Conclusion:The results indicated that there was a significant shift in the ruminal microflora and metabolites associated with inflammation and immune responses during CM.Moreover,in the rumen of cows affected by SM,the relative abundance of several opportunistic pathogens and the level of metabolites which could produce antibacterial compounds or had a competitive inhibitory effect were all increased.

Month-Wise Prevalence of Subclinical Mastitis in Dairy Cows in Guangdong Province,China

To evaluate the month-wise prevalence of subclinical mastitis （SM） and its relations with climatic temperature and humidity, and to provide references for control and prevention of mastitis in dairy cows in Guangdong, China, California mastitis test method was used to determine the monthly prevalence of SM （MPSM, %） of lactating Holstein cows from 11 dairy farms in Guangdong Province, China, including about 516 cows per monthly examination. The average MPSM on a cow and a quarter basis were 30 and 13% respectively, and there was a positive linear correlation between monthly mean air temperature （MT, °C） and MPSM on a cow basis （r=0.763, P=0.004） or a quarter basis （r=0.577, P=0.049）, but there were no correlations between MT and MPSM on a cow or a quarter basis （P〉0.05）. There was a shift in MPSM trend that the highest MPSM （38.4%） was not for the hottest July with MT at 30.8°C, but for the later September at 27.9°C. The farms need to develop new housing conditions to control cowshed temperatures in the hot season to reduce the mastitis prevalences in the post-hot months.

In-vitro antimicrobial activity screening of some ethnoveterinary medicinal plants traditionally used against mastitis,wound and gastrointestinal tract complication in Tigray Region,Ethiopia

Objective:To screen the antibacterial activity of nine ethnoveterinary plants traditionally used for the treatment of mastitis,wound and gastrointestinal complications.Methods:Hydroalcoholic exctracts of medicinal plants namely,Achyranthes aspera(A.aspera) L.(Family Asparagaceae),Ficus caria(F.caria)(Family Moraceae).Malvi parviflora(M.parviflora)(Family Malvaceae),Vernonia species(V.species) docal name Alakit.Family Asteraceae),Solatium hastifolium(S.hastifolium)(Family Solanaceae),Calpurinia aurea(C.aurea)(Ait) Benth(Family Fabaceae),Nicotiana tabacum(N.tabacum) L.(Family Solanaceae),Ziziphus spina-christi(Z.spina-christi)(Family Rhamnaceae).Croton macroslachys(C.macrostachys)(Family Fuphorhiaceae).were screened against clinical bacterial isolates of veterinary importance from October 2007 to April2009.The antibacterial activity was tested using disc diffusion at two concentrations(200 mg/mL and 100 mg/mL) and broth dilution methods using 70%methanol macerated leal extracts.Results:With the exception ol S.hastifolium all plant extracts exhibited antibacterial activity.Among the medicinal plants tested C.aurea.C.macrostachyus,A.aspera,N.tabacum and vcrnonia species(Alakit) showed the most promising antimicrobial properties.Conclusions:It can be concluded that many of the tested plants have antibacterial activity and supports the traditional usage of the plants for mastitis,wound and gastrointestinal complications treatment.Further studies into their toxicity and phytochemistry is advocated.

Relationship between granulomatous lobular mastitis and methylene tetrahydrofolate reductase gene polymorphism

BACKGROUND Variations in the methylene tetrahydrofolate reductase(MTHFR)gene have been reported as risk factors for numerous conditions,including cardiovascular disease,thrombophilia,stroke,hypertension and pregnancy-related complications.Moreover,it was reported there is an association between breast cancer and mutations in MTHFR-C677T.However,whether there is an association between MTHFR gene polymorphism and granulomatous lobular mastitis or not has been rarely investigated.AIM To analyze the association between MTHFR gene polymorphism and granulomatous lobular mastitis.METHODS Fifty-one patients with granulomatous lobular mastitis admitted to The First Hospital of Kunming were selected as study samples.Their hospitalization time ranged from February 2018 to February 2019.The 51 patients were included in the experimental group,and another 51 women who underwent physical examination at The First Hospital of Kunming in the same period were included in the control group.Deoxyribonucleic acid and MTFR genetic polymorphism testing were performed in each group.The association between MTHFR gene polymorphism and granulomatous lobular mastitis was observed.RESULTS There were significant differences in genotype frequency and allele frequency of C/C and C/T between the experimental group and the control group(all P<0.05).However,there was no significant difference in frequency of T/T genotype between the two groups(P>0.05).In addition,there was no significant difference in genotype frequency and allele frequency of A/A,A/C and C/C between the two groups(P>0.05).CONCLUSION MTHFR gene C677T locus polymorphism is closely related to granulomatous lobular mastitis.

Feeding a Saccharomyces cerevisiae fermentation product improves udder health and immune response to a Streptococcus uberis mastitis challenge in mid-lactation dairy cows

Background:We aimed to characterize the protective effects and the molecular mechanisms of action of a Saccharomyces cerevisiae fermentation product(NTK)in response to a mastitis challenge.Eighteen mid-lactation multiparous Holstein cows(n=9/group)were fed the control diet(CON)or CON supplemented with 19 g/d NTK for 45 d(phase 1,P1)and then infected in the right rear quarter with 2500 CFU of Streptococcus uberis(phase 2,P2).After 36-h,mammary gland and liver biopsies were collected and antibiotic treatment started until the end of P2(9 d post challenge).Cows were then followed until day 75(phase 3,P3).Milk yield(MY)and dry matter intake(DMI)were recorded daily.Milk samples for somatic cell score were collected,and rectal and udder temperature,heart and respiration rate were recorded during the challenge period(P2)together with blood samples for metabolite and immune function analyses.Data were analyzed by phase using the PROC MIXED procedure in SAS.Biopsies were used for transcriptomic analysis via RNA-sequencing,followed by pathway analysis.Results:DMI and MY were not affected by diet in P1,but an interaction with time was recorded in P2 indicating a better recovery from the challenge in NTK compared with CON.NTK reduced rectal temperature,somatic cell score,and temperature of the infected quarter during the challenge.Transcriptome data supported these findings,as NTK supplementation upregulated mammary genes related to immune cell antibacterial function(e.g.,CATHL4,NOS2),epithelial tissue protection(e.g.IL17C),and anti-inflammatory activity(e.g.,ATF3,BAG3,IER3,G-CSF,GRO1,ZFAND2A).Pathway analysis indicated upregulation of tumor necrosis factorα,heat shock protein response,and p21 related pathways in the response to mastitis in NTK cows.Other pathways for detoxification and cytoprotection functions along with the tight junction pathway were also upregulated in NTK-fed cows.Conclusions:Overall,results highlighted molecular networks involved in the protective effect of NTK prophylactic supplementation on udder health during a subclinical mastitic event.

Polysaccharide Nucleic Acid of Bacillus Calmette Guerin Modulates Th1/Th2 Cytokine Gene Expression in Lipopolysaccharide-Induced Mastitis in Rats

The aim of this study was to evaluate, in rats, the changes in the T helper type 1（Th1）/Th2 radio in mammary glands after an intramammary infusion of lipopolysaccharide （LPS） and to characterize the moderating effects of the polysaccharide nucleic acid of Bacillus Calmette Guerin （BCG-PSN） on the mammary gland. In the control group, the levels of IL-2 and INF-7 mRNA expression increased, whereas IL-4 mRNA expression decreased after LPS challenge. As a consequence, the INF-γ/IL-4 mRNA ratio was significantly higher at 3, 6, and 9 h post-infusion （PI） compared to the control value （0 h; P〈0.01）. BCG-PSN increased mRNA expression of both INF-γ and IL-4 before infusion of LPS. LPS challenge significantly the reduced Th1/Th2 cytokine ratio due to Thl cytokine IFN-γ suppression and Th2 cytokine IL-4 upregulation compared with the control group. A significant reduction of N-acetyl-β-D-glucosaminidase （NAGase） was observed at 24 h PI in the BCG-PSN treatment group compared to the control group （P〈 0.05）. Thus, it was demonstrated that level of BCG-PSN might change the Th1/Th2 ratio mainly by enhancing the Th2 immune response. This is the first report of a Th1/Th2 change induced by coliform mastitis and characterization of the effect of BCG-PSN on mammary gland inflammation. This study makes a better understanding of the mechanisms of coliform mastitis and provides a putative novel strategy for the prevention and/or treatment of mastitis.