地衣型真菌次级代谢产物抗菌活性初步研究

地衣是一种传统的民族药物,能产生多种具有活性的物质。该研究对地衣型真菌(Xanthoria elegans,Myelochroa indica,Ramalina peruviana,Cladonia macilenta,Nephromopsis pallescens,Cladonia coccifera)进行液体培养,2个月后,培养液用乙酸乙酯萃取后获得初提物。该研究采用抑菌圈法评价地衣型真菌初提物对7种致病细菌(Bacillus subtilis,Bacillus cereus,Vibrio parahaemolyticus,Straphylococcus haemolyticus,Pseudomonas aeruginosa,Staphylococcus aureus,Micrococcus luteus)的抗菌活性,并测定最低抑菌浓度(MIC)。结果表明:6种地衣型真菌的初提物均具有一定的抗菌活性,且不同培养基对地衣型真菌产生抗菌物质有显著影响。其中,R.peruviana在MY液体培养基中所产生的次级代谢产物对金黄色葡萄球菌、藤黄微球菌、溶血性葡萄球菌、铜尿假单胞菌具有抑制效果,但在YMG培养基中所得初提物对供试7种致病细菌不具有抑菌效果。X.elegans在YMG培养基中所得初提物对枯草芽孢杆菌具有明显抗菌活性,其抑菌圈直径可达17.77 mm。该研究证实不同地衣型真菌液体培养初提物具有抗菌活性,不同的培养基也直接影响地衣型真菌抗菌效果。该研究结果为地衣型真菌的进一步研究及民族药的开发利用奠定了基础。     

Diversity and exploration of bioactive marine actinomycetes in the Bay of Bengal of the Puducherry coast of India

The present study was designed to investigate the Puducherry coast of the Bay of Bengal, India for the diversity of bioactive actinomycetes. A total of 50 actinomycete strains were isolated from the marine sediments and most of the strains were belongs to Streptomyces. These strains were identified by means of morphological physiological, biochemical and cultural characteristics. The isolates were subjected to shake flask fermentation and the secondary metabolites were extracted with ethyl acetate and screened for cytotoxicity, hemolytic activity and antimicrobial activity against selected bacterial and fungal pathogens. The cytotoxic activity was evaluated using HeLa cell lines by 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide, a tetrazole (MTT) assay, hemolytic activity on mouse erythrocytes and the antifungal activity was evaluated by MTT cytotoxic assay against Aspergillus niger, Aspergillus fumigatus and Candida albicans. The antibacterial activity was studied against Staphylococcus aureus, Bacillus subtilis, Escherichia coli and Klebsiella pneumoniae. The cytotoxicity and antimicrobial activity of secondary metabolite was found to be concentration dependent and nearly 24% of isolates showed significant antimicrobial, hemolytic and cytotoxic activity. The results of our study indicate the diversity and bioactive potential of marine actinomycetes isolated in the Puducherry coast.     

Phytochemical, antimicrobial, antioxidant and antigenotoxic potentials of Cyperus rotundus extracts

The aqueous, ethyl acetate, methanolic and Total Oligomer Flavonoids (TOF) enriched extracts, obtained from the aerial parts of Cyperus rotundus, were investigated for their contents in phenolic compounds. Antioxidative activity using the NBT/riboflavin assay system, antimicrobial activity against Gram positive and Gram negative bacterial reference strains as well as antigenotoxic activity tested with the SOS chromotest assay were also studied. Significant antibacterial activity against reference strains; Staphylococcus aureus, Enterococcus faecalis, Salmonella enteritidis and Salmonella typhimurium, was detected in the presence of ethyl acetate and TOF enriched extracts. In addition to their antimicrobial activity, the same extracts showed a significant ability to inhibit nitroblue tetrazolium reduction by the superoxide radical in a non enzymatic O₂^.− generating system, and were also able to reduce significantly the genotoxicity induced by nifuroxazide and Aflatoxin B1. The antioxidant, antimicrobial and antigenotoxic activities exhibited by C. rotundus depend on the chemical composition of the tested extracts.     

Molecular profiling of marine endophytic fungi from green algae: Assessment of antibacterial and anticancer activities

Bioactive natural metabolites, especially from the marine endophytic fungi, are largely unexplored. Endophytic fungi are being increasingly recognized as a group of organisms that produce novel metabolites of industrial importance. This study investigated the anticancer and antibacterial potential of the marine algal endophyte, Penicillium chrysogenum. The different organic solvent extracts of the endophytic fungi grown on different growth medium were analyzed for anticancer and antibacterial activities. The highest inhibitory activity was observed for the ethyl acetate (EA) extract of the culture filtrate grown in potato dextrose broth (PDB) for 21 days, against the tested human breast cancer cell (MCF-7) line. Similarly, the PDB-EA extract showed an appreciable activity against the human pathogens. The biochemical analysis of the Cha EA metabolites revealed terpenoids, steroids, phenolics and flavones. Gas Chromatography (GCMS) data revealed several bioactive compounds such as anthraquinone and cinnamic acid. The Cha EA extract induced membrane damage and thus, apoptosis in MCF-7cells. The secondary metabolites produced by these marine endophytic fungi have contributed to considerable anticancer and antimicrobial activities and hence, this study is an evidence of potential sources of antimicrobial and anticancer compounds from Penicillium chrysogenum.     

Antibacterial and antilarval-settlement potential and metabolite profiles of novel sponge-associated marine bacteria

In this study, we screened seven novel sponge-associated marine bacteria for their antibacterial and antilarval-settlement activity in order to find possible new sources of non-toxic or less toxic bioactive antifoulants. The anti-bacterial-growth activity of crude extracts of each bacterium was evaluated by the disk-diffusion assay. Extracts of four potent bacteria with high and broad spectra of antibacterial activity were further separated with solvents of different polarities (hexane and ethyl acetate). To evaluate their indirect inhibitive effect on larval settlement, we tested for their antibiofilm formation activity against two of the test bacteria (Vibrio halioticoli and Loktanella hongkongensis) inductive to Hydroides elegans larval settlement. About 60 and 87% of the extracts inhibited biofilm formation by V. halioticoli and by L. hongkongensis respectively. The extracts were also tested for their direct antilarval-settlement activity against the barnacle Balanus amphitrite and the polychaete H. elegans; 87% of the extracts had a strong inhibitive effect on larval settlement of both species. Extracts of two of the isolates completely inhibited larval settlement of B. amphitrite at 70 μg ml⁻¹ and H. elegans at 60 μg ml⁻¹. The organic extracts of Winogradskyella poriferorum effectively inhibited the larval settlement of both H. elegans and B. amphitrite and the biofilm formation of the two bacterial species. The metabolites present in the active crude extracts were profiled using GC MS, and the most prevalent metabolites present in all extracts were identified. This study successfully identified potential new sources of antifouling compounds.

The study of antagonistic interactions among pelagic bacteria: a promising way to coin environmental friendly antifouling compounds

Ten strains of marine bacteria (SCH0401–SCH0410) were isolated from Ayajin, the east coast of South Korea. In spectrophotometer based chemotaxis assay the ethyl acetate extract (300 μg) of SCH0402 decreased the optical density (OD) of the motile target strains SCH0401, SCH0402, SCH0407 and SCH0408 by two to six times when compared to control. Tributyltin oxide (TBTO) decreased the OD of all target strains by only two times. The most active strain SCH0402 was identified as Shewanella oneidensis by using 16S rDNA gene sequence analysis. Similarly, the target motile strains SCH0401, SCH0402, SCH0407 and SCH0408 were identified as Alteromonas marina, Shewanella oneidensis, Roseobacter gallaeciensis and Bacillus atrophaeus, respectively. The growth inhibition zone produced by the test bacterial extracts against the target strains were three to eight times smaller when compared to that of TBTO. Even though, SCH0402 showed six times weaker antibacterial activity, the repellent activity was three times stronger than TBTO. Therefore, the higher negative chemotactic activity would be better to select eco-friendly antifouling compounds than the other antibacterial activities.     

Influence of glucose and oxygen on the production of ethyl acetate and isoamyl acetate by a <Emphasis Type="Italic">Saccharomyces cerevisiae</Emphasis> strain during alcoholic fermentation

The effect of glucose and dissolved oxygen in a synthetic medium simulating the standard composition of grape juice on the production of ethyl acetate and isoamyl acetate by a Saccharomyces cerevisiae strain during alcoholic fermentation was studied. The specific in vitro activity of alcohol acetyltransferase (AATase, EC 2.3.1.84) and esterases (ESase, EC 3.1.1.1; hydrolysis and synthesis of esters) in cell-free extracts was also examined. The specific activity of AATase for ethyl acetate was found to peak at the beginning of the exponential growth phase and that for isoamyl acetate at its end. While the glucose concentration only affected the maximum specific activity of AATase, and only slightly, oxygen inhibited such activity, to a greater extent for isoamyl acetate than for ethyl acetate. On the other hand, esterases were found to catalyse the synthesis of ethyl acetate only at a low or medium glucose concentration (50 or 100 g l^-1, respectively), and to reach their maximum hydrolytic activity on isoamyl acetate during the stationary growth phase. The highest ethyl acetate and isoamyl acetate concentrations in the medium were obtained with a glucose concentration of 250 g l^-1 and semianaerobic conditions.     

Bionectria ochroleuca NOTL33—an endophytic fungus from Nothapodytes foetida producing antimicrobial and free radical scavenging metabolites

Endophytic fungi are reported to produce diverse classes of secondary metabolites. This study investigated the antimicrobial and free radical scavenging activity of a foliar endophytic fungus from Nothapodytes foetida, a medium sized tree known to produce the antineoplastic compound camptothecin. The fungal isolate was identified as Bionectria ochroleuca based on the ITS rDNA analysis. The differences among endophytic, pathogenic and free living Bionectria ochroleuca were established by RNA secondary structure analysis. The metabolites showed a broad spectrum of antibacterial, antifungal and anti-dermatophytic activity. Minimum inhibitory concentration values of ethyl acetate extracts were in the range of 78–625 μg/mL against all test organisms, except for Pseudomonas aeruginosa (5 mg/mL). Antimicrobial components in the ethyl acetate extract were identified by GC-MS analysis. The isolate was also produced volatile antifungal compounds. A dose-dependent free radical quenching was observed in the ethyl acetate extract. This is the first report on Bionectria sp. as an endophyte of N. foetida. The results indicate that the B. ochroleuca NOTL33 isolate is a potential source of antimicrobial agents and could be used as an effective biofumigant.     

Optimization of process parameters for improved production of bioactive metabolite by a novel endophytic fungus <Emphasis Type="Italic">Fusarium</Emphasis> sp. DF2 isolated from <Emphasis Type="Italic">Taxus wallichiana</Emphasis> of North East India

An endophytic fungus having antifungal and antibacterial properties was isolated from Taxus wallichiana of Arunachal Pradesh, India. On the basis of morphological and molecular characteristics, the fungus was identified as Fusarium sp. and designated as DF2. The fungus was optimized for growth and maximum production of the antimicrobial agent. Media with 5% leaf extract (w/v) supplemented with 0.1% dextrose as carbon and yeast extract as nitrogen source favored the growth with temperature optimum 25 ± 2°C and pH 6. Incubation period of 10 days was observed optimum for growth and production of antimicrobial agent. Phenylalanine and dextrose enriched basal medium promoted the antimicrobial agent production, whereas methionine amended in combination with glucose promoted higher biomass accumulation. The TLC purified active compound with UV λ-max 270 nm in ethyl acetate has got the lowest minimum inhibitory concentration (MIC) against Bacillus subtilis, Staphylococcus aureus and Escherichia coli and highest against Pseudomonas aeruginosa.     

Activity of South African medicinal plants against Listeria monocytogenes biofilms,and isolation of active compounds from Acacia karroo

In South Africa, the antimicrobial activity of many indigenous plants has been investigated. In general, studies have focused on planktonic bacteria, with less attention given to bacterial biofilms. Many organisms, however, including the opportunistic pathogen Listeria monocytogenes occur more frequently as biofilms. The aim of this study was to identify and select plants that exhibit the best antilisterial activity, isolate the bioactive compounds and determine their effect on the architecture of listerial biofilms. The ethyl acetate and chloroform extracts of thirteen plants were investigated for antilisterial activity. The ethyl acetate extract of Acacia karroo and Plectranthus ecklonii showed the best antilisterial activity, exhibiting a minimum inhibitory concentration (MIC) of 3.1 mg/ml and 0.5 mg/ml, respectively. These were further selected for the identification of bioactive compounds. Column chromatographic purification of the ethyl acetate extracts of the leaves of A. karroo led to the isolation of three known pure compounds, namely epicatechin (1), β-sitosterol (2) and epigallocatechin (3). Confocal scanning laser microscopy (CSLM) showed that the biomass of the listerial biofilm was reduced when the isolated compounds were added. The aggregation of cells that were exposed to β-sitosterol and epigallocatechin was reduced from 25 μm as observed in untreated cells to < 10 μm in length.     

中华婪步甲共生放线菌的抗菌活性筛选及菌株BJ37的初步鉴定

利用平板分离法从中华婪步甲(Harpalus sinicus)中分离得到56株共生放线菌。其中菌株BJ37具有较好的抗菌活性,其发酵液对枯草芽胞杆菌和白色念珠菌的抑菌圈直径均大于30.0 mm。进一步研究BJ37发酵液不同极性溶剂萃取物的抗菌活性,结果表明在中等极性部位存在活性物质。通过形态学特征和分子生物学分析确定该菌株相似菌株为委内瑞拉链霉菌(Streptomyces venezuelae)。菌株BJ37作为微生物杀菌剂潜在产生菌值得进一步研究。     

Optimization and production of novel antimicrobial agents from sponge associated marine actinomycetes <Emphasis Type="Italic">Nocardiopsis dassonvillei</Emphasis> MAD08

The sponge-associated actinomycetes were isolated from the marine sponge Dendrilla nigra, collected from the southwest coast of India. Eleven actinomycetes were isolated depending upon the heterogeneity and stability in subculturing. Among these, Nocardiopsis dassonvillei MAD08 showed 100% activity against the multidrug resistant pathogens tested. The culture conditions of N. dassonvillei MAD08 was optimized under submerged fermentation conditions for enhanced antimicrobial production. The unique feature of MAD08 includes extracellular amylase, cellulase, lipase, and protease production. These enzymes ultimately increase the scope of optimization using broad range of raw materials which might be efficiently utilized. The extraction of the cell free supernatant with ethyl acetate yielded bioactive crude extract that displayed activity against a panel of pathogens tested. Analysis of the active thin layer chromatography fraction by Fourier transform infrared and gas chromatography-mass spectrometry evidenced 11 compounds with antimicrobial activity. The ammonium sulfate precipitation of the culture supernatant at 80% saturation yielded an anticandidal protein of molecular weight 87.12 kDa. This is the first strain that produces both organic solvent and water soluble antimicrobial compounds. The active extract was non-hemolytic and showed surface active property envisaging its probable role in inhibiting the attachment of pathogens to host tissues, thus, blocking host–pathogen interaction at an earlier stage of pathogenesis.     

The Diversity and Antimicrobial Activity of Preussia sp. Endophytes Isolated from Australian Dry Rainforests

Limited knowledge currently exists regarding species diversity and antimicrobial activity of endophytic isolates of Preussia within Australia. This report describes endophytic Preussia species that were identified through molecular analysis of the internal transcribed spacer region. Screening for antimicrobial secondary metabolites was determined by testing crude ethyl acetate (EtOAc) extracts derived from fungal mycelia against a panel of ATCC type strains which included Bacillus cereus, Escherichia coli, Enterococcus faecalis, Pseudomonas aeruginosa, Serratia marcescens, methicillin-resistant Staphylococcus aureus (MRSA) and the opportunist yeast pathogen Candida albicans. Subsequently, high-performance liquid chromatography generated fractions of bioactive EtOAc extracts which were subject to confirmatory testing using the Clinical and Laboratory Standards Institute reference microdilution antimicrobial activity assay. A total of 18 Preussia were isolated from nine host plants with 6/18 having a <97 % sequence similarity to other known species in Genbank, suggesting that they are new species. In preliminary screening, 13/18 Preussia isolates revealed antimicrobial activity against at least one of the microbes tested, whilst 6/18 isolates, including 4/6 putative new species showed specific antimicrobial activity against MRSA and C. albicans. These results highlight the antimicrobial potential of Australian Preussia spp. and also the importance of Australian dry rainforests as an untapped repository of potentially significant bioactive compounds.     

Composition and biological activities of different extracts of Salvia jurisicii,a rare and endemic Macedonian species

This is the first report on the phenolic composition and biological activities of endemic species Salvia jurisicii Ko?anin (Lamiaceae) originating from Macedonia. Aerial parts of S. jurisicii were extracted with dichloromethane, ethyl acetate, methanol, ethanol and water. All extracts were tested spectrophotometrically for total phenolic and flavonoid contents, while their phenolic composition was analyzed using HPLC-DAD. The antioxidant activity of extracts was studied using 2,2-dyphenyl-1-picrylhydrazyl, 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid), ferric reducing ability of plasma and ß-carotene bleaching assays. Being the most frequently used solvents for extraction of bioactive ingredients from medicinal plants, water and ethanol extracts were chosen for further testing of antimicrobial, cytotoxic and antineurodegenerative activities. More polar solvent extracts showed higher total phenolic content and lower flavonoid content. Ethanol and methanol extracts, followed by water extract exhibited the strongest antioxidant activity. Extracts showed antimicrobial activity against certain bacteria and micromycetes. The cytotoxicity assay showed slight toxicity to HCT-116 cell line. In the antineurodegenerative assays, the extracts performed lower inhibition of acetylcholinesterase and tyrosinase than standards. According to the results of this study, it can be inferred that more polar extracts of S. jurisicii were quantitatively richer in total phenolics, showing stronger antioxidant activity. Ethanol extract showed stronger biological activities comparing to water extract.     

Purification and characterization of antibacterial substances produced by a marine bacterium Pseudoalteromonas haloplanktis strain

Aims: To purify and characterize compounds with antimicrobial activity from Pseudoalteromonas haloplanktis inhibition (INH) strain. Methods and Results: The P. haloplanktis isolated from a scallop hatchery was used to analyse antibacterial activities. Crude extracts were obtained with ethyl acetate of the cultured broth, after separation of bacterial cells, and assays against six strains of marine bacteria and nine clinically important pathogenic bacteria. The active compounds were purified from ethyl acetate extracts, by a combination of SiO₂ column and thin layer chromatography. Two active fractions were isolated, and chemical structures of two products from the major one were unambiguously identified as isovaleric acid (3-methylbutanoic acid) and 2-methylbutyric acid (2-methylbutanoic acid), by comparing their mass spectra and ¹H- and ¹³C-nuclear magnetic resonance spectra to those of authentic compounds. Conclusions: In the antibacterial activity of P. haloplanktis INH strain, extra cell compounds are involucred, mainly isovaleric and 2-methylbutyric acids. Significance and Impact of the Study: Production of antimicrobial compounds by marine micro-organisms has been widely reported; however, the efforts not always are conducted to purification and applications of these active compounds. This study is a significant contribution to the knowledge of compounds unique from marine bacteria as potential sources of new drugs in the pharmacological industry.     

桦褐孔菌提取物抗氧化活性研究

以乙酸乙酯和甲醇为提取剂,采用索氏提取法,剩余残渣采用热水浸提,最终得到桦褐孔菌不同极性提取物,对其DPPH自由基、羟基自由基及超氧阴离子自由基清除活性作用进行了研究,确定桦褐孔菌的抗氧化能力,为深入研究和开发桦褐孔菌功能性食品奠定理论基础。实验结果表明桦褐孔菌具有较好的抗氧化活性,其中乙酸乙酯提取物的DPPH自由基清除率、羟基自由基清除率和超氧阴离子自由基清除率均高于其他两组分及BHT,桦褐孔菌提取物有望成为功能性食品组分中合成抗氧化剂的天然替代品。     

Antimicrobial potential of a lipopeptide biosurfactant derived from a marine Bacillus circulans

Aims: To isolate the biologically active fraction of the lipopeptide biosurfactant produced by a marine Bacillus circulans and study its antimicrobial potentials. Methods and Results: The marine isolate B. circulans was cultivated in glucose mineral salts medium and the crude biosurfactant was isolated by chemical isolation method. The crude biosurfactants were solvent extracted with methanol and the methanol extract was subjected to reverse phase high‐performance liquid chromatography (HPLC). The crude biosurfactants resolved into six major fractions in HPLC. The sixth HPLC fraction eluting at a retention time of 27·3 min showed the maximum surface tension‐reducing property and reduced the surface tension of water from 72 mNm^?1 to 28 mNm^?1. Only this fraction was found to posses bioactivity and showed a pronounced antimicrobial action against a panel of Gram‐positive and Gram‐negative pathogenic and semi‐pathogenic micro‐organisms including a few multidrug‐resistant (MDR) pathogenic clinical isolates. The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of this antimicrobial fraction of the biosurfactant were determined for these test organisms. The biosurfactant was found to be active against Gram‐negative bacteria such as Proteus vulgaris and Alcaligens faecalis at a concentration as low as 10 μg ml^?1. The biosurfactant was also active against methicillin‐resistant Staphylococcus aureus (MRSA) and other MDR pathogenic strains. The chemical identity of this bioactive biosurfactant fraction was determined by post chromatographic detection using thin layer chromatography (TLC) and also by Fourier transform infrared (FTIR) spectroscopy. The antimicrobial HPLC fraction resolved as a single spot on TLC and showed positive reaction with ninhydrin, iodine and rhodamine‐B reagents, indicating its lipopeptide nature. IR absorption by this fraction also showed similar and overlapping patterns with that of other lipopeptide biosurfactants such as surfactin and lichenysin, proving this biosurfactant fraction to be a lipopeptide. The biosurfactant did not show any haemolytic activity when tested on blood agar plates, unlike the lipopeptide biosurfactant surfactin produced by Bacillus subtilis. Conclusions: The biosurfactant produced by marine B. circulans had a potent antimicrobial activity against Gram‐positive and Gram‐negative pathogenic and semi‐pathogenic microbial strains including MDR strains. Only one of the HPLC fractions of the crude biosurfactants was responsible for its antimicrobial action. The antimicrobial lipopeptide biosurfactant fraction was also found to be nonhaemolytic in nature. Significance and impact of the study: This work presents a nonhaemolytic lipopeptide biosurfactant produced by a marine micro‐organism possessing a pronounced antimicrobial action against a wide range of bacteria. There is a high demand for new antimicrobial agents because of the increased resistance shown by pathogenic micro‐organisms against the existing antimicrobial drugs. This study provides an insight into the search of new bioactive molecules from marine micro‐organisms.     

杜仲内生真菌DZ05的鉴定及抗菌生物活性研究

结合菌株的形态学特征和ITS序列分析结果,对1株分离自杜仲茎部的内生真菌菌株DZ05进行鉴定,并对其在PDA液体培养基摇床培养3 d获得的发酵液对多种测试菌进行抗菌活性研究。结果显示:(1)分离自杜仲茎部的内生真菌菌株DZ05经形态学特征和ITS序列分析,被鉴定为淡紫色拟青霉。(2)其发酵液的乙酸乙酯提取物对6种测试细菌和9种测试植物病原菌均具有明显的抑菌活性,抑菌圈直径在13～45 mm之间,其中对番茄灰霉病菌、番茄叶霉病菌和苹果炭疽病菌抑菌圈直径＞40 mm。研究表明,杜仲内生真菌DZ05的代谢产物具有广谱的抗菌活性,在植物病原菌的生物防治领域具有较大的应用前景。     

Optimization and production of pyrrolidone antimicrobial agent from marine sponge-associated Streptomyces sp. MAPS15

Twenty-nine actinobacterial strains were isolated from marine sponge Spongia officinalis and screened for antagonistic activity against various bacterial and fungal pathogens. The active antibiotic producer MAPS15 was identified as Streptomyces sp. using 16S rRNA phylogenetic analysis. The critical control factors were selected from Plackett–Burman (PB) factorial design and the bioprocess medium was optimized by central composite design (CCD) for the production of bioactive metabolite from Streptomyces sp. MAPS15. The maximum biomass and active compound production obtained with optimized medium was 6.13 g/L and 62.41 mg/L, respectively. The economical carbon source, paddy straw was applied for the enhanced production of bioactive compound. The purified active fraction was characterized and predicted as pyrrolidone derivative which showed broad spectrum of bioactivity towards indicator organisms. The predicted antimicrobial spectra suggested that the Streptomyces sp. MAPS15 can produce a suite of novel antimicrobial drugs.     

Antimicrobial activity and phytochemical screening of Ocimum americanum L extracts against pathogenic microorganisms

The aim of the present study is to assess the antimicrobial activities of various leaf extracts of Ocimum americanum were tested against pathogenic microorganisms. Preparation of different extracts viz., aqueous, acetone, ethyl acetate and methanol through soxhlet extraction method. Various extracts were investigated against MTCC strains of Bacillus cereus, Clostridium penfrigens, Klebsilla pnemoniae, Salmonella paratyphi, Candida albicans and Aspergillus niger by agar well diffusion and disc diffusion methods. Minimum inhibitory concentration (MIC), Minimum Bactericidal/Fungicindal Concentration (MBC/MFC) were determined through micro dilution method. Elucidation of phytochemicals and functional groups were observed by HPLC and FT-IR respectively. Ethyl acetate leaf extract of O.americanum showed significant antimicrobial activity against the all tested pathogens in agar well diffusion method in which B.cereus (17 mm) was observed high zone of inhibition. Whereas lowest inhibition was observed in aqueous extract against C.pentrigens (7 mm). The ranges of MIC values from 0.78 μg/ml to 50 μg/ml and MBC/MFC 1.56 μg/ml to 50 μg/ml were observed. Phytochemicals such as alkaloids, steroids, saponins, flavonoids, tannins, terepenes, phenolic compounds cardiac glycosides were detected. Saponinns, flavonoids, tannins, phenolic compounds were observed in only ethyl acetate leaf extracts. Functional group of the leaf extracts was exhibited by FTIR and HPLC analysis of the ethyl acetate leaf extract was elutated at six peaks. Based on the results we concluded that ethyl acetate leaf extract of O.americanum has proved to be potentially effective than the other extracts. Therefore, ethyl acetate leaf extract of O.americanum could act as antimicrobial agent and further studies are recommended for isolation of compounds and toxicological studies.