Isolation and identification of multidrug-resistant Staphylococcus haemolyticus from a laboratory-breeding mouse

Objective:To analysis and identify a bacterium strain isolated from laboratory breeding mouse far away from a hospital.Methods:Phenotype of the isolate was investigated by conventional microbiological methods,including Gram-staining,colony morphology,tests for haemolysis, catalase,coagulase,and antimicrobial susceptibility test.The meek and 16S rRNA genes were amplified by the polymerase chain reaction(PCR) and sequenced.The base sequence of the PCR product was compared with known 16S rRNA gene sequences in the CenBank database by phylogenetic analysis and multiple sequence alignment.Results:The isolate in this study was a gram positive,coagulase negative,and catalase positive coccus.The isolate was resistant to oxacillin,methicillin,penicillin,ampicillin,cefazolin,cipr of loxacin erythromycin,et al.PCR results indicated that the isolate was meek gene positive and its 16S rRNA was 1465 bp.Phylogenetic analysis of the resultant 16S rRNA indicated the isolate belonged to genus Saphylococcus,and multiple sequence alignment showed that the isolate was Saphylococcus haemolyticus with only one base difference from the corresponding 16S rRNA deposited in the CenBank.Conclusions: 16S rRNA gene sequencing is a suitable technique for non-specialist researchers.Laboratory animals are possible sources of lethal pathogens,and researchers must adapt protective measures when they manipulate animals.

Inhibition of quorum sensing in drug-resistant Staphylococcus epidermidis by flavaspidic acid BB

Background:To investigate the effect of flavaspidic acid BB(BB)on biofilms and quorum sensing-related genes of drug-resistant Staphylococcus epidermidis(SE)and to provide a theoretical basis for the development of BB as a new type of quorum sensing inhibitor.Methods:The microdilution method was applied to screen 15 clinical isolates of S.epidermidis for drug-resistant S.epidermidis to be used as the test strain.The effects of BB on the biofilms of drug-resistant S.epidermidis at different growth stages were studied using the cell counting kit-8 assay and scanning electron microscopy.The gene expression of sigB,sarA,and luxS,regulators involved in quorum sensing of biofilm formation,was measured by PCR.Results:The minimum inhibitory concentrations of BB against 15 clinical strains of S.epidermidis ranged 11.67-66.67μg/mL.BB significantly inhibited biofilm formation by drug-resistant SE06 in the aggregation and maturation stages,with an activity superior to those of mupirocin and fusidic acid(P<0.05).At 40μg/mL,BB reduced the secretion of extracellular polymers and promoted the disruption of biofilm structure.At 80μg/mL,BB specifically interfered with the quorum sensing system of drug-resistant SE06 by downregulating sarA and luxS,thereby inhibiting this microbe’s biofilm formation.Conclusion:BB exhibited strong inhibitory activity against SE06 by suppressing biofilm formation and downregulating quorum sensing-related genes and was identified as a new potential quorum sensing inhibitor against S.epidermidis.

A Review of <i>Staphylococcus aureus</i>and the Emergence of Drug-Resistant Problem

There are various bacteria living in this world. The most common one is Staphylococcus aureus. Almost everyone has heard of it. It is easy to find their habitats, such as hospitals, homes, parks, schools etc. Some of them are difficult to be eliminated because of drug-resistant mutations. Hence, lots of researchers devoted their efforts to eliminate them. This review illustrates the characteristics of the Staphylococcus aureus and the main threat of their drug-resistant strains, especially methicillin-resistant S. aureus. What’s more, the article also highlights the plight in the drug development.

Metal nanoparticles as inhibitors of enzymes and toxins of multidrug-resistant Staphylococcus aureus

Staphylococcus aureus is an aerobic Gram-positive spherical bacterium known to cause a broad range of infections worldwide.It is a major cause of infective skin and soft infections and severe and life-threatening conditions,such as pneumonia,bloodstream infections,and endocarditis.The emergence of drug-resistant strains of S aureus,par-ticularly methicillin-resistant S aureus(MRSA),has become a significant concern in the healthcare community.Antibiotic-resistant S aureus is commonly acquired in hospitals and long-term care facilities.It often affects pa-tients with weakened immune systems,those undergoing invasive medical procedures,or those who have been hospitalized for extended periods.In the US,S aureus is known to cause potentially fatal illnesses,such as toxic shock syndrome(TSS)and acute-onset toxic shock syndrome(TSS),which are characterized by fever and hy-potension.It develops resistance to antibiotics through several mechanisms,such as the production of enzymes that inactivate antibiotics,target site modification,efflux pumps,and plasmid-mediated resistance.Therefore,preventing the spread of drug-resistant S aureus is needed,and there is an urgent need to explore novel ap-proaches in the development of anti-staphylococcal agents.This article reviews the principal infections caused by S aureus,major virulence factors,mechanisms of resistance development,and nanotechnology-based solutions for the control of drug-resistant S aureus.

甘草提取物对Staphylococcus aureus的抑菌活性及作用机理

食源性病菌为食品安全带来了巨大挑战,甘草提取物作为食品添加剂对金黄色葡萄球菌(Staphylococcus aureus)具有良好的抑菌作用,可作为天然食品防腐剂的候选原料,但目前对其抑菌机理的研究还不深入,影响了其应用。为探究甘草提取物对S.aureus的抑菌机理,该研究通过生长曲线、氧化损伤实验、细胞膜壁分析、蛋白质分析和DNA分析,评价了甘草提取物对S.aureus的抑菌作用机制。结果表明,甘草提取物导致S.aureus核酸渗漏,说明其膜完整性被破坏;同时,甘草提取物降低了几种能量代谢酶:琥珀酸脱氢酶(succinate dehydrogenase,SDH)和总ATP酶的活力;此外,光谱和竞争分析表明,甘草提取物与DNA发生了静电结合和凹槽结合。总之,甘草提取物主要是通过对S.aureus细胞壁膜、蛋白质合成、细菌代谢活力和遗传物质发挥作用,从而抑制其生长。该研究为甘草提取物在食品防腐方面的应用提供了理论基础。

Community-acquired multidrug-resistant pneumonia,bacteraemia,and infective endocarditis:A case report

BACKGROUND The prevalence of multidrug-resistant(MDR)bacteria has increased globally,with extensive drug-resistant(XDR)bacteria posing a threat to patients.CASE SUMMARY This case report describes a young man admitted for suspected tropical fever infections who experienced rapid deterioration in health.Despite negative results for tropical fever infections,he had neutrophilic leucocytosis,acute kidney injury,and chest imaging findings suggestive of bilateral consolidations.On day two,he was diagnosed with infective endocarditis with possible rheumatic heart disease and MDR methicillin-resistant Staphylococcus aureus bacteraemia,and communityacquired pneumonia.Despite treatment with broad-spectrum antibiotics,he did not respond and succumbed to death on day five.CONCLUSION This case highlights that clinicians/public should be aware of MDR communityacquired pneumonia,bacteraemia,and endocarditis which ultimately culminate in high rates of morbidity and mortality.Early identification of pathogenic strain and prompt antibiotic treatment are a mainstay for the management and prevention of early fatalities.Simultaneously,route cause analysis of communityacquired MDR/XDR pathogens is a global need.

Global burden of HIV-negative multidrug-and extensively drug-resistant tuberculosis based on Global Burden of Disease Study 2021

Background:Tuberculosis(TB),caused by Mycobacterium tuberculosis,remains the second leading cause of death from a single infectious disease globally and poses a significant economic and clinical burden in the world in 2022.Of particular concern is the emergence of drug-resistant TB,accounting for 15%-20%of TB deaths.It is imperative to delve into the global trends of incidence and death rate for multidrug-resistant tuberculosis(MDRTB)and extensively drug-resistant tuberculosis(XDR-TB),drawing upon the comprehensive Global Burden of Disease(GBD)2021 drug-resistant tuberculosis dataset.Methods:From the GBD 2021,data on incidence,prevalence,disability-adjusted life years(DALYs),and death of MDR-TB and XDR-TB from 1990 to 2021 were collected.We calculated the estimated annual percentage changes in age standardized incidence rate(ASIR)and age-standardized death rate(ASDR),segmented by age,sex,and socio-demographic index(SDI).The impacts of various risk factors on MDR-TB and XDR-TB were also analyzed.Results:In 2021,there were an estimated 443,680(95%uncertainty interval[UI]:259,196-766,545)incident cases of MDR-TB,and an estimated 106,818(95%UI:41,612-211,854)death cases of MDR-TB,while there were an estimated 24,036(95%UI:17,144-34,587)incident cases of XDR-TB and 7,946(95%UI:3,326-14,859)death cases of XDR-TB.The incidence and death cases of MDR-TB were lowest in high SDI regions,whereas the incidence rates of XDR-TB in high-middle SDI regions were higher than those in middle SDI and high SDI regions.Conclusion:This study reported the disease burden of drug-resistant TB from 1990 to 2021.Until 2021,drugresistant TB is still a serious problem in low SDI countries,especially for high-risk age populations with highrisk factors.Controlling drug-resistant TB requires effective control strategies and healthcare systems.

金黄色葡萄球菌(Staphylococcus aureus)液体增菌培养基的优化

为研发一种金黄色葡萄球菌(Staphylococcus aureus)选择性增菌液,选用针对革兰氏阴性菌及非葡萄球菌属菌株具有选择抑制作用的7.5%氯化钠肉汤作为基础培养基,探索氨基酸、丙酮酸钠、甘露醇对金黄色葡萄球菌的促生长能力,通过正交试验确定培养基的最优配方,并使用优化后的培养基与原始培养基作用于来源不同的四株金黄色葡萄球菌标准菌株,对其增菌效果和抑制大肠杆菌(Escherichia coli)的效果进行对比。结果表明,最佳培养基配方为在基础培养基上添加L-丙氨酸1.0 g/L、甘露醇6.0 g/L、丙酮酸钠1.2 g/L。菌株在优化后培养基上增菌浊度及转接至Baird-Parker的菌落数上均有促进效果,最终菌株的OD550 nm值有0.28~0.34的增长,转接Baird-Parker后的菌落数也有10~100倍的增长,且在促进目标菌生长的同时对大肠杆菌的抑制性也没有受到影响,表明该增菌培养基选择性良好。

Staphylococcus aureus and biofilms:transmission, threats, and promising strategies in animal husbandry

Staphylococcus aureus(S. aureus) is a common pathogenic bacterium in animal husbandry that can cause diseases such as mastitis, skin infections, arthritis, and other ailments. The formation of biofilms threatens and exacerbates S. aureus infection by allowing the bacteria to adhere to pathological areas and livestock product surfaces, thus triggering animal health crises and safety issues with livestock products. To solve this problem, in this review, we provide a brief overview of the harm caused by S. aureus and its biofilms on livestock and animal byproducts(meat and dairy products). We also describe the ways in which S. aureus spreads in animals and the threats it poses to the livestock industry. The processes and molecular mechanisms involved in biofilm formation are then explained. Finally, we discuss strategies for the removal and eradication of S. aureus and biofilms in animal husbandry, including the use of antimicrobial peptides, plant extracts, nanoparticles, phages, and antibodies. These strategies to reduce the spread of S. aureus in animal husbandry help maintain livestock health and improve productivity to ensure the ecologically sustainable development of animal husbandry and the safety of livestock products.

Structural insights on anti-biofilm mechanism of heated slightly acidic electrolyzed water technology against multi-resistant Staphylococcus aureus biofilm on food contact surface

Slightly acidic electrolyzed water(SAEW)has proven to be an efficient and novel sanitizer in food and agriculture field.This study assessed the efficacy of SAEW(30 mg/L)at 40℃on the inactivation of foodbome pathogens and detachment of multi-resistant Staphylococcus aureus(MRSA)biofilm.Furthermore.the underlying mechanism of MRS A biofilm under heated SAEW at 40℃treatment on metabolic profiles was investigated.The results showed that the heated SAEW at 40℃significantly effectively against foodbome pathogens of 1.96-7.56(lg(CFU/g))reduction in pork,chicken,spinach,and lettuce.The heated SAEW at 40℃treatment significantly reduced MRS A biofilm cells by 2.41(lg(CFU/cm^(2))).The synergistic effect of SAEW treatment showed intense anti-biofilm activity in decreasing cell density and impairing biofilm cell membranes.Global metabolic response of MRSA biofilms,treated by SAEW at 40℃,revealed the alterations of intracellular metabolites,including amino acids,organic acid,fatty acid,and lipid.Moreover,signaling pathways involved in amino acid metabolism,energy metabolism,nucleotide synthesis,carbohydrate metabolites,and lipid biosynthesis were functionally disrupted by the SAEW at 40℃treatment.As per our knowledge,this is the first research to uncover the potential mechanism of heated SAEW treatment against MRSA biofilm on food contact surface.

Elimination of methicillin‑resistant Staphylococcus aureus biofilms on titanium implants via photothermally‑triggered nitric oxide and immunotherapy for enhanced osseointegration

Background:Treatment of methicillin-resistant Staphylococcus aureus(MRSA)biofilm infections in implant placement surgery is limited by the lack of antimicrobial activity of titanium(Ti)implants.There is a need to explore more effective approaches for the treatment of MRSA biofilm infections.Methods:Herein,an interfacial functionalization strategy is proposed by the integration of mesoporous polydopamine nanoparticles(PDA),nitric oxide(NO)release donor sodium nitroprusside(SNP)and osteogenic growth peptide(OGP)onto Ti implants,denoted as Ti-PDA@SNP-OGP.The physical and chemical properties of Ti-PDA@SNP-OGP were assessed by scanning electron microscopy,X-ray photoelectron spectroscope,water contact angle,photothermal property and NO release behavior.The synergistic antibacterial effect and elimination of the MRSA biofilms were evaluated by 2′,7′-dichlorofluorescein diacetate probe,1-N-phenylnaphthylamine assay,adenosine triphosphate intensity,O-nitrophenyl-β-D-galactopyranoside hydrolysis activity,bicinchoninic acid leakage.Fluorescence staining,assays for alkaline phosphatase activity,collagen secretion and extracellular matrix mineralization,quantitative real‑time reverse transcription‑polymerase chain reaction,and enzyme-linked immunosorbent assay(ELISA)were used to evaluate the inflammatory response and osteogenic ability in bone marrow stromal cells(MSCs),RAW264.7 cells and their co-culture system.Giemsa staining,ELISA,micro-CT,hematoxylin and eosin,Masson's trichrome and immunohistochemistry staining were used to evaluate the eradication of MRSA biofilms,inhibition of inflammatory response,and promotion of osseointegration of Ti-PDA@SNP-OGP in vivo.Results:Ti-PDA@SNP-OGP displayed a synergistic photothermal and NO-dependent antibacterial effect against MRSA following near-infrared light(NIR)irradiation,and effectively eliminated the formed MRSA biofilms by inducing reactive oxygen species(ROS)-mediated oxidative stress,destroying bacterial membrane integrity and causing leakage of intracellular components(P<0.01).In vitro experiments revealed that Ti-PDA@SNP-OGP not only facilitated osteogenic differentiation of MSCs,but also promoted the polarization of pro-inflammatory M1 macrophages to the anti-inflammatory M2-phenotype(P<0.05 or P<0.01).The favorable osteo-immune microenvironment further facilitated osteogenesis of MSCs and the anti-inflammation of RAW264.7 cells via multiple paracrine signaling pathways(P<0.01).In vivo evaluation confirmed the aforementioned results and revealed that Ti-PDA@SNP-OGP induced ameliorative osseointegration in an MRSA-infected femoral defect implantation model(P<0.01).Conclusions:Ti-PDA@SNP-OGP is a promising multi-functional material for the high-efficient treatment of MRSA infections in implant replacement surgeries.

The ever-changing microenvironment of Staphylococcus aureus in cutaneous infections

Background:Staphylococcus aureus is responsible for the majority of skin and soft tissue infections,which are often diagnosed at a late stage,thereby impacting treatment efficacy.Our study was designed to reveal the physiological changes at different stages of infection by S.aureus through the combined analysis of variations in the skin microenvironment,providing insights for the diagnosis and treatment of S.aureus infections.Methods:We established a murine model of skin and soft tissue infection with S.aureus as the infectious agent to investigate the differences in the microenvironment at different stages of infection.By combining analysis of the host immune status and histological observations,we elucidate the progression of S.aureus infection in mice.Results:The results indicate that the infection process in mice can be divided into at least two stages:early infection(1–3 days post-i nfection)and late infection(5–7 days post-i nfection).During the early stage of infection,notable symptoms such as erythema and abundant exudate at the infection site were observed.Histological examination revealed infiltration of numerous neutrophils and bacterial clusters,accompanied by elevated levels of cytokines(IL-6,IL-10).There was a decrease in microbial alpha diversity within the microenvironment(Shannon,Faith's PD,Chao1,Observed species,Simpson,Pielou's E).In contrast,during the late stage of infection,a reduction or even absence of exudate was observed at the infected site,accompanied by the formation of scabs.Additionally,there was evidence of fibroblast proliferation and neovascularization.The levels of cytokines and microbial composition gradually returned to a healthy state.Conclusion:This study reveals synchrony between microbial composition and histological/immunological changes during S.aureus-i nduced SSTIs.

Eugenol targeting CrtM inhibits the biosynthesis of staphyloxanthin in Staphylococcus aureus

Staphylococcus aureus is a serious foodborne pathogen threatening food safety and public health.Especially the emergence of methicillin-resistant Staphylococcus aureus(MRSA)increased the difficulty of S.aureus treatment.Staphyloxanthin is a crucial virulence factor of S.aureus.Blocking staphyloxanthin production could help the host immune system counteract the invading S.aureus cells.In this study,we first screened for staphyloxanthin inhibitors using a virtual screening method.The outcome of the virtual screening method resulted in the identification of eugenol(300μg/mL),which significantly inhibits the staphyloxanthin production in S.aureus ATCC 29213,S.aureus Newman,MRSA ATCC 43300 and MRSA ATCC BAA1717by 84.2%,63.5%,68.1%,and 79.5%,respectively.The outcome of the growth curve assay,field-emission scanning electron,and confocal laser scanning microscopy analyses confirmed that eugenol at the test concentration did not affect the morphology and growth of S.aureus.Moreover,the survival rate of S.aureus ATCC 29213 and MRSA ATCC 43300 under H_(2)O_(2) pressure decreased to 51.9%and 45.5%in the presence of eugenol,respectively.The quantitative RT-PCR and molecular simulation studies revealed that eugenol targets staphyloxanthin biosynthesis by downregulating the transcription of the crtM gene and inhibiting the activity of the CrtM enzyme.Taken together,we first determined that eugenol was a prominent compound for staphyloxanthin inhibitor to combat S.aureus especially MRSA infections.

Electrochemical and colorimetric dual-signal detection of Staphylococcus aureus enterotoxin B based on AuPt bimetallic nanoparticles loaded Fe-N-C single atom nanocomposite

Sensitive detection of Staphylococcus aureus enterotoxin B(SEB)is of importance for preventing food poisoning from threatening human health.In this work,an electrochemical and colorimetric dual-signal detection assay of SEB was developed.The probe(Ab2/AuPt@Fe-N-C)was bound to SEB captured by Ab1,where the Ab2/AuPt@Fe-N-C triggered methylene blue degradation and resulted in the decrease of electrochemical signal.Furthermore,the probe catalyzed the oxidation of 3,3’,5,5’-tetramethyl biphenyl to generate a colorimetric absorbance at 652 nm.Once the target was captured and formed a sandwich-like complex,the color changed from colorless to blue.SEB detection by colorimetric and electrochemical methods showed a linear relationship in the concentration ranges of 0.0002-10.0000 and 0.0005-10.0000 ng/mL,with limits of detection of 0.0667 and 0.1670 pg/mL,respectively.The dual-signal biosensor was successfully used to detect SEB in milk and water samples,which has great potential in toxin detection in food and the environment.

Community-acquired pneumonia:The importance of the early detection of drug-resistant organisms

Pneumonia is a disease associated with significant healthcare burden with over 1.5 million hospitalizations annually and is the eighth leading cause of death in the United States.While community-acquired pneumonia(CAP)is generally considered an acute time-limited illness,it is associated with high long-term mortality,with nearly one-third of patients requiring hospitalization dying within one year.An increasing trend of detecting multidrug-resistant(MDR)organisms causing CAP has been observed,especially in the Western world.In this editorial,we discuss about a publication by Jatteppanavar et al which reported that a case of a MDR organism was the culprit in developing pneumonia,bacteremia,and infective endocarditis that led to the patient’s death.The early detection of these resistant organisms helps improve patient outcomes.Significant advances have been made in the biotechnological and research space,but preventive measures,diagnostic techniques,and treatment strategies need to be developed.

Antimicrobial and synergistic effects of lemongrass and geranium essential oils against Streptococcus mutans,Staphylococcus aureus,and Candida spp.

BACKGROUND The oral cavity harbors more than 700 species of bacteria,which play crucial roles in the development of various oral diseases including caries,endodontic infection,periodontal infection,and diverse oral diseases.AIM To investigate the antimicrobial action of Cymbopogon Schoenanthus and Pelargonium graveolens essential oils against Streptococcus mutans,Staphylococcus aureus,Candida albicans,Ca.dubliniensis,and Ca.krusei.METHODS Minimum microbicidal concentration was determined following Clinical and Laboratory Standards Institute documents.The synergistic antimicrobial activity was evaluated using the Broth microdilution checkerboard method,and the antibiofilm activity was evaluated with the 3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide assay.Data were analyzed by one-way analysis of variance followed by the Tukey post-hoc test(P≤0.05).RESULTS C.schoenanthus and P.graveolens essential oils were as effective as 0.12%chlorhexidine against S.mutans and St.aureus monotypic biofilms after 24 h.After 24 h P.graveolens essential oil at 0.25%was more effective than the nystatin group,and C.schoenanthus essential oil at 0.25%was as effective as the nystatin group.CONCLUSION C.schoenanthus and P.graveolens essential oils are effective against S.mutans,St.aureus,Ca.albicans,Ca.dubliniensis,and Ca.krusei at different concentrations after 5 min and 24 h.

葡萄球菌Staphylococcus condimenti对广式腊肠风味的影响

采用固相微萃取方法结合气质联用仪,对Staphylococcus condimenti发酵广式腊肠中风味物质成分进行测定,分析接种分离菌株对广式腊肠风味的影响。从对照组和接种组(SC)中均鉴定出22种风味物质成分,主要为醇类、醛类、酮类和酯类物质,还有部分烃类和环类化合物。研究表明,Staphylococcus condiment可以加速脂肪和蛋白质的降解与氧化,有助于腊肠风味的改善。

Cloning and Prokaryotic Expression of FnBP Ligand Binding Gene of Staphylococcus aureus

[Objective] The study aimed to clone the FnBP ligand binding gene of Staphylococcus aureus and run prokaryotic expression by constructing a prokaryotic expression vector. [Method] The gene encoding FnBP ligand binding gene was amplified from S.aureus chromosomal DNA by PCR technique. After T-A cloning, plasmid pMD18- FnBP was constructed. pMD18- FnBP and pET28a(+)were digested by BamH Ⅰ and EcoR Ⅰ double enzymes, then the purified FnBP ligand binding gene was subcloned into the expression vector pET28a(+), and the prokaryotic expression vector pET28a-FnBP was thus constructed. The constructed plasmid pET28a-FnBP was transformed into Escherichia coli BL21(DE3) competent cells. The bacterium was induced by IPTG and the expressed products were analyzed by SDS-PAGE and Western blot. [Result] The gene fragment with the length of 370 bp was amplified by PCR approach. One approximately 30 kD exogenous protein was observed in SDS-PAGE analysis. Western blot analysis indicates the protein has antigenicity of S.aureus. [Conclusion] The FnBP ligand binding gene of S.aureus was successfully cloned and expressed in prokaryotic cells.

The Effect of Culture Condition on Type 5 Capsular Polysaccharide Production of Staphylococcus aureus from Diary Cattle

[Objective] The effect of different culture conditions on type 5 capsular polysaccharide production of Staphylococcus aureus from diary cattle was studied to provide simple way for CP production and preparation and laid foundation for carrying out new polysaccharide vaccine research. [Method] Staph-ylococcus aureus was isolated from milk sample of sick dairy cattle and capsular polysaccharide serotypes were identified. Type 5 capsular polysaccharide was cultured on BHI,solid columbia and mod110 culture media. Glucose and lactose were taken as carbon sources for every culture media in solid and liquid state. Therefore 9 different culture conditions were taken to study the effect of culture conditions on capsular polysaccharide production. [Result] Different culture conditions indicated that compared with columbia culture media, BHI culture media could decline capsular polysaccharide production and mod110 culture media could increase capsular polysaccharide production. While for same culture media, solid culture media was better for capsular polysaccharide production,meanwhile,taken lactose as carbon source could increase capsular polysaccharide production.

Inhibitory Effects of Bacillus subtilis on Staphylococcus aureus

[Objective] To produce drug resistance, seek non-toxic environmental so as to change the current biological drugs that did not excessive use of antibiotics. [Methods] A strain of Bacillus was purified and isolated from fresh and healthy in- testines of grass carps. Biochemical identification was carried out by conventional bacterial biochemical test method. Two pairs of primers were designed, 16S rRNA detection and sequencing analysis were carried out. Drug sensitive test was carried out by agar diffusion method. In vitro inhibition test on Staphylococcus aureus was carried out by Oxford cup method. [Results] The isolated bacterium had basically the same biochemical characters as Bacillus subtilis; and the homology reached 100%. Thus, the isolated bacterium was identified to be Bacillus subtilis. It was insensitive to amoxicillin, ampicillin, penicillin G and so on, but sensitive to amikacin, cefalexin, ciprofloxacin and cefradine. The inhibitory effects of Bacillus subtilis on Staphylococ- cus aureus were significant. The minimum inhibitory concentration （MIC） was 2.8×10^8×2^-5/ml and minimum bactericidal concentration （MBC） was 2.8×10^8×2^-2/ml. [Conclusions] The isolated Bacillus subtilis could be used to prevent and control diseases caused by Staphylococcus aureus, and reduce the abuse of antibiotics.