控制多重耐药微生物的抗菌药物生物合成新试验

M. Elaasser
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摘要

本研究试图从当地生境土壤微生物群中寻找一种新的产药微生物,以控制多重耐药问题。抗菌素耐药性(AMR)这个术语是用来描述微生物能够抵抗旨在杀死它们的药物和化学物质的作用。从埃及15个不同地区的土壤样品中分离到76株放线菌,采用琼脂塞扩散法对被试微生物进行了初步的抗菌活性筛选。选取16株分离菌株,在小规模深层发酵系统中进行二次筛选,并用琼脂孔扩散法对病原微生物进行检测。其中,分离物(S1SHA1)对病原菌的抗菌活性最高。通过形态学、生理生化特征和16s rRNA基因序列鉴定该分离物为灰纹链霉菌。研究了影响抗菌剂活性的物理和营养因素。结果表明,当pH为7,培养温度28℃,搅拌150 rpm,碳氮源淀粉1.5%,硝酸钾0.4%,磷2 g/l, NaCl浓度为1%时,抑菌剂活性最佳。采用乙酸乙酯法和制备层析法分别对间歇培养的抗菌剂进行提取和纯化。透射电镜(TEM)测定稻瘟病球菌(S. griseoplanus, S1SHA1)抑菌剂对试验微生物菌株的最低抑菌浓度(MIC)和作用方式。细胞毒性研究表明,即使在高浓度测试时,也没有观察到该化合物的细胞毒性作用
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A NEW TRIAL FOR BIOFORMATION OF ANTIMICROBIAL AGENT CONTROLLING MULTI DRUG RESISTANT MICROORGANISM
In the present investigation, a trial was done to find a new antimicrobial agent producing microbe from soil microbiota of local habitats to control the problem of multiple drug resistance. The term of Antimicrobial resistance (AMR) is used to describe microorganisms that can resist the effects of drugs and chemicals designed to kill them. Seventy six actinomycetes isolates were isolated from fifteen soil samples different localities in Egypt were primary screening for antimicrobial activity by agar plug diffusion method against test microorganisms. Sixteen isolates were selected for secondary screening in small scale submerged fermentation system and assayed against pathogenic tested microorganisms using agar well diffusion method. Among of these isolates tested, the isolate (S1SHA1) showed the highest antimicrobial activity against pathogenic test organisms. This isolate was identified as Streptomyces griseoplanus by morphological, physiological, biochemical characters and 16s rRNA gene sequence. Physical and nutritional factors affecting activity of antimicrobial agent were studied. The results showed that, optimum activity of antimicrobial agent achieved with pH 7, incubation temperature 28 C, for 7 days, at 150 rpm agitation, carbon and nitrogen source starch 1.5% and potassium nitrate 0.4%, as well as phosphorus 2 g/l and NaCl at concentration of 1 %. Antimicrobial agent from batch culture was subjected to extraction and purification processes using ethyle acetate and preparative TLC, respectively. Determination of minimum inhibitory concentrations (MIC) and Mode of action of antimicrobial agent produced by S. griseoplanus (S1SHA1) on the test microbial strains using Transmission Electron Microscopy (TEM). Cytotoxic studies showed that no cytotoxic effects were observed for the compound when tested even at high concentrations
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