淡水振荡蓝藻粗提物体外抗菌、抗真菌及细胞毒活性的筛选。

W. Wijesekara, P. Manage
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引用次数: 4

摘要

蓝藻是一种高度多样化的原核生物,被认为是具有抗病毒、抗菌、抗真菌和抗癌特性的生物活性化合物的有效来源。本研究的目的是筛选来自斯里兰卡Senanayaka Samudraya水库(70 0 11’37.37 0 N 810 31’47.13 0 E)的淡水蓝藻振荡藻胞内次级代谢物的抗菌、抗真菌和细胞毒活性。采用琼脂盘扩散法筛选振荡菌对革兰氏阳性耐甲氧西林金黄色葡萄球菌(MRSA) ATCC 25923、炭疽芽孢杆菌和革兰氏阴性铜绿假单胞菌(ATCC 25853)、伤寒沙门菌和大肠杆菌(ATCC 25922)、真菌、单细胞白色念珠菌(ATCC 60192)和热带念珠菌的体外抗菌和抗真菌活性。测定了振荡藻粗提物的最小抑菌浓度(MIC)、最小杀菌浓度(MBC)、最小杀真菌浓度(MFC)和细胞毒作用(盐水对虾生物测定)。用己烷和甲醇分别提取10%和60%的生物质。采用气相色谱-质谱联用技术(GC-MS)对粗提物中的化合物进行鉴定。粗提物甲醇提取物对金黄色葡萄球菌(19±2 mm)和白色念珠菌(10±1mm)在24 h内的抑菌活性最高,己烷提取物仅对金黄色葡萄球菌(19±2 mm)有抑菌活性,24 h内的平均抑菌带直径为11±1mm。甲醇提取物对金黄色葡萄球菌的最低MIC为156.25µg/ml。甲醇提取物对金黄色葡萄球菌和白色念珠菌的最低MBC和MFC分别为0.63 mg/ml和1.25mg/ml。测定50%粗提物对盐水对虾的致死浓度为2.50 g/l、1.25 g/l和0.625 g/l,间隔6、12、24 h。气相色谱-质谱分析结果表明,该植物甲醇粗提物中含有重要脂肪酸:十六烷酸甲酯、甲基十四烷和十三烷酸,正己烷提取物中含有二(2-乙基己基羟基吡啶氧化物、1 -苯二甲酸单(2-乙基己基)酯、邻苯二甲酸6-乙基-3-辛基庚基酯和邻苯二甲酸十二烷基壬基酯,具有抗菌和抗真菌作用。关键词:振荡菌,MFC, MIC, MBC,细胞毒性
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In vitro screening of, antibacterial antifungal and cytotoxicity activities in crude extract of freshwater cyanobacterium Oscillatoria sp.
Cyanobacteria, highly diverse group of prokaryotes are recognized as a potent source of biologically active compounds with antiviral, antibacterial, antifungal, and anticancer properties.The aim of the present study was to screen antibacterial, antifungal and cytotoxic activities of intracellular secondary metabolites of freshwater cyanobacterium Oscillataria sp. Cyanobacterium Oscillatoria sp. was isolated from Senanayaka Samudraya reservoir (7 0 11’ 37.37 0 N 810 31’ 47.13 0 E), Sri Lanka.In vitro antibacterial and antifungal activity of Oscillatoria sp. was screened against Gram-positive Methicillin-resistant Staphylococcus aureus (MRSA) ATCC 25923, Bacillus anthracis and Gram-negative Pseudomonas aeruginosa (ATCC 25853), Salmonella typhi and Escherichia coli (ATCC 25922) and fungi, unicellular Candida albicans (ATCC 60192) and Candida tropicalis using agar disc diffusion method.The Minimum Inhibitory Concentrations (MIC), Minimum Bacteriocidal Concentration (MBC), Minimum Fungicidal Concentrations (MFC) and cytotoxic effects (brine shrimp bioassay) of Oscillatoria crude extract were determined. 10% and 60% of biomass was extracted with hexane and methanol respectively.Gas Chromatography-Mass Spectrometry (GC-MS) was used to identify compounds in the crude extract.  The highest antibacterial and antifungal activity of crude extract were detectedin methanol extract against S.aureus (19 ± 2 mm)and C.albicans (10 ± 1 mm) within 24 hours wherein the hexane extract, antibacterial activity was detected only for S. aureus and mean diameter of inhibition zone was 11 ± 1mm within 24 hours.The lowest MIC of methanol extract against S. aureus wasfound as 156.25 µg/ml. The lowest MBC and MFC of methanol extract against S. aureus and C.albicans were 0.63 mg/ml and 1.25mg/ml respectively. Lethal concentration, 50% of the crude extract against brine shrimp was recorded at 2.50  g/l, 1.25 g/l and 0.625 g/l for 6, 12, 24 hrs intervals respectively. GC-MS analysis revealed that the methanol crude extract of Oscillatoria sp. contains important fatty acid namely hexadecanoic acid methyl ester, methyl tetradecane and 13-tetradecanoic acid and n-hexane extract contains Bis (2-ethylhexyl hydroxypyridine oxide, 1 2-benzenedicarboxylic acid mono (2-ethylhexyl) ester, Phthalic acid 6-ethyl-3-Octyl heptyl ester and  Phthalic acid  dodecyl nonyl ester which may possess antibacterial  and antifungal properties. Keywords: Oscillatoria sp., MFC, MIC, MBC, Cytotoxicity
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