Quantitative analysis of quercetin content of Blumea Balsamifera L. DC Dichloromethane leaf fraction using HPLC-RP-PDA and direct TLC-Bioautography

@#Quercetin, a flavonoid compound which is widely distributed in plants are considered ass beneficial physiologically due to attributed bioactivity such as anti-cancer, immunomodulatory, antidiabetic, and anti-inflammatory. In this study, the quercetin content from the dried Blumea balsamifera L. DC dried leaf was macerated with 95% ethanol and the concentrated extract was purified using Modified Kupchan method and flash chromatography. All fractions were tested for the presence of flavonoids using phytochemical screening and the selected dichloromethane fraction were further purified using another round of flash chromatograph. All resulting fractions and pooled samples were tested for the antioxidant property using the developed Thin Layer Chromatography (TLC)-Bioautography and separated compounds were derivatized with DPPH. Using the optimized TLC-Bioautography method, the quercetin content in the dichloromethane fraction was analyzed and compared with a reversed phase high performance liquid chromatography hyphenated with photodiode array detector (RP-HPLC-PDA). The calculated quercetin content from the pooled sample using TLC-bioautography method is 2.25 mg/ml and from RP-HPLC-PDA is 2.02 mg/ml which was not comparable statistically using unpaired t-test (p<0.05, α=0.05

Partial purification and characterization of antimicrobial peptide(s) from Mimosa pudica (Mimosaceae)

@#Plants have been a major source of natural products for sustaining human health. The use of the different parts of the plant as infusions, decoctions, extracts, and powders are being employed in the treatment of different diseases in humans, plants, and animals. One property of great significance in terms of therapeutic treatments, especially with the emergence of multi-drug resistant microbes, is the antimicrobial activity. A new promising source of antimicrobials that demonstrate novel mechanisms of therapeutic strategies is low molecular weight peptides. In this study, the antimicrobial activities of Mimosa pudica crude and partially purified peptide extracts against Gram-negative Enterobacter cloacae ATCC 23355 and Enterobacter aerogenes ATCC 13048, and Gram-positive Staphylococcus epidermidis ATCC 12228 using resazurin colorimetric assay and tricine SDS-PAGE bioautography were reported. M. pudica crude and partially purified extracts exhibited antimicrobial activity against all the bacteria tested. Specifically, the peptide that was partially purified from M. pudica with a molecular weight of 5.14 kDa inhibited the growth of Enterobacter cloacae.

Anti-Acinetobacter baumannii activity of Rumex crispus L. and Rumex sanguineus L. extracts

Objective: To examine the effect of Rumex crispus (R. crispus) and Rumex sanguineus (R. sanguineus) plant extracts against isolates of Acinetobacter baumannii (A. baumannii) from wounds, including multidrug-resistant strains.Methods: Six prepared Rumex extracts were subjected to liquid chromatography-tandem mass spectrometry. Antimicrobial activity of extracts and pure compounds (catechin, quercetin, isoquercitrin, emodin, and gallic acid) was examined by a microtiter plate method, while for determination of compound binary combinations activity a checkerboard method was applied. Active fractions of extracts were detected by agar-overlay high-performance thin-layer chromatography-bioautography assay followed by liquid chromatography - diode array detection - mass spectrometry analysis. Results: A total of 28 compounds were detected in two extracts of R. crispus and 26 compounds in four different R. sanguineus extracts, with catechin as a dominant component. Anti-A. baumannii activity was confirmed for all six R. sanguineus and R. crispus extracts at the concentration range from 1 to 4 mg/mL. Neither examined single compounds nor their binary combinations exhibited an anti-A. baumannii activity (MIC>256 μg/mL). The bioautography showed that fractions with the most prominent anti-A. baumannii activity tended to contain more polar compounds, predominantly flavonol (quercetin and kaempherol) glycosides; but also fractions containing flavanone (eriodictyol) glycosides and anthraquinone (emodin) glycosides; and less polar eriodictyol aglycone. Conclusions: The results justify and elucidate the traditional application of R. sanguineus and R. crispus extracts for wound healing, indicating the necessity for their further examination in combat against multidrug-resistant A. baumannii isolates from wounds.

Anti-Acinetobacter baumannii activity of Rumex crispus L. And Rumex sanguineus L. extracts

Objective: To examine the effect of Rumex crispus (R. crispus) and Rumex sanguineus (R. sanguineus) plant extracts against isolates of Acinetobacter baumannii (A. baumannii) from wounds, including multidrug-resistant strains. Methods: Six prepared Rumex extracts were subjected to liquid chromatography-tandem mass spectrometry. Antimicrobial activity of extracts and pure compounds (catechin, quercetin, isoquercitrin, emodin, and gallic acid) was examined by a microtiter plate method, while for determination of compound binary combinations activity a checkerboard method was applied. Active fractions of extracts were detected by agar-overlay high-performance thin-layer chromatography-bioautography assay followed by liquid chromatography-diode array detection-mass spectrometry analysis. Results: A total of 28 compounds were detected in two extracts of R. crispus and 26 compounds in four different R. sanguineus extracts, with catechin as a dominant component. Anti-A. baumannii activity was confirmed for all six R. sanguineus and R. crispus extracts at the concentration range from 1 to 4 mg/mL. Neither examined single compounds nor their binary combinations exhibited an anti-A. baumannii activity (MIC>256 μg/mL). The bioautography showed that fractions with the most prominent anti-A. baumannii activity tended to contain more polar compounds, predominantly flavonol (quercetin and kaempherol) glycosides; but also fractions containing flavanone (eriodictyol) glycosides and anthraquinone (emodin) glycosides; and less polar eriodictyol aglycone. Conclusions: The results justify and elucidate the traditional application of R. sanguineus and R. crispus extracts for wound healing, indicating the necessity for their further examination in combat against multidrug-resistant A. baumannii isolates from wounds. Aleksic Sabo Verica 1 Department of Biology and Ecology, Faculty of Sciences, University of Novi Sad, Trg Dositeja Obradovica 3, 21 000 Novi Sad, Vojvodina Svircev Emilija 2 Department of Chemistry, Biochemistry and environmental protection, Faculty of Sciences, University of Novi Sad, Trg Dositeja Obradovica 3, 21 000 Novi Sad, Vojvodina Mimica-Dukic Neda 3 Department of Chemistry, Biochemistry and environmental protection, Faculty of Sciences, University of Novi Sad, Trg Dositeja Obradovica 3, 21 000 Novi Sad, Vojvodina Orcic Dejan 4 Department of Chemistry, Biochemistry and environmental protection, Faculty of Sciences, University of Novi Sad, Trg Dositeja Obradovica 3, 21 000 Novi Sad, Vojvodina Narancic Jelena 5 Department of Biology and Ecology, Faculty of Sciences, University of Novi Sad, Trg Dositeja Obradovica 3, 21 000 Novi Sad, Vojvodina Knezevic Petar 6 Department of Biology and Ecology, Faculty of Sciences, University of Novi Sad, Trg Dositeja Obradovica 3, 21 000 Novi Sad, Vojvodina Almasaudi SB. 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Antimicrobial Assay of Aqueous Extracts of Selected Ethno-pharmacologic Alternatives Used by the Maasai Community of Narok, Kenya

Aims: Antimicrobial resistance motivates the search for new antimicrobials. Besides Methicillin-Resistant Staphylococcus aureus, Carbapenem-Resistant Klebsiella pneumoniae strain has emerged worldwide over the last decade, posing a great challenge to healthcare. This paper reports a survey of Maasai ethno-pharmacy practices. Study Design: Key informant interviews and utilization of e-questionnaires for data collection. Methodology: Plants were identified, and the applicable parts taken as samples, dried, powdered then subjected to aqueous extraction. Using agar well diffusion method, the extracts were screened against gram positive, gram negative and fungal strains to establish antimicrobial activity. Place and Duration of Study: The study was conducted at the School of Pharmacy & Health Sciences of the United States International University, Africa in Nairobi from January 2017 to December 2018. Results: Out of the 24 different plant samples collected, 33% were leaves while 17%, 12.5% and 37.5% were fruits, stem bark and roots, respectively. The highest extract percentage yields were from the leaves of Biden pilosa (5.11%), Psidium guajava (4.65%) and Tarchononthus comphoratus (4.31%). While the minimum extracts yields were from Solanum incum roots (0.08%) and stem bark (0.09%). The extracts of Toddalia asiatica stem bark and roots; Rhamnus staddo roots; Tarchonanthus camphoratus stem bark and roots; and Zanthroxyleum chelybeum stem bark, all exhibited well defined inhibition diameters against M.R.S. aureus in the range 8mm to 14mm as compared to the standard drug (10mm). All these were extracts of non-leafy samples. The significant antimicrobial activity corresponded to presence of flavonoids and alkaloids as seen on TLC plates during phytochemical screening. Conclusion: The results obtained are a good rationale for utilization of the plants identified as alternatives to antibiotics for management of antimicrobial infections.

Solvent-solvent fractionations of Combretum erythrophyllum (Burch.) leave extract: Studies of their antibacterial, antifungal, antioxidant and cytotoxicity potentials

OBJECTIVE@#To evaluate the biological activities of Combretum erythrophyllum (C. erythrophyllum) leaf extracts against infectious diseases' pathogenesis and their cytotoxicity potentials.@*METHODS@#Powdered leaf material (300 g) of C. erythrophyllum was extracted (1:10 w/v) using acetone to obtain the crude extract. Liquid-liquid fractionation was performed on the crude acetone extract (30 g) using solvents of different polarity. The bioautographic method was used to detect the inhibition of bacterial and fungal growth by active compounds present in the crude and fractions. The extracts were then tested on bacterial strains: Staphylococcus aureus, Enterococcus faecalis, Escherichia coli, Pseudomonas aeruginosa; fungal strains: Candida albicans (C. albicans), Cryptococcus neoformans, and Aspergillus fumigatus, by microtitre dilution method for MIC determination.@*RESULTS@#The extracts MIC values ranged between 0.08 and 2.50 mg/mL against the tested pathogens. Water fraction had the highest activity against bacteria strains, while the fungal assay revealed crude acetone extract and ethyl acetate fraction to be active against C. albicans (1.25 mg/mL), dichloromethane extract against C. albicans and A. fumigatus (0.16 mg/mL). Extract fractions showed a good antioxidant activity via DPPH, ABTS and hydroxyl radical scavenging assays, in the order: ethyl acetate > water > acetone > dichloromethane > hexane. The toxicity level of crude extract and fractions evaluated in Vero monkey kidney cells ranged from 34 to 223 μg/mL, while doxorubicin (IC = 7.19 μg/mL) served as the positive control.@*CONCLUSIONS@#It can be concluded that the extracts of C. erythrophyllum are safe for medicinal use in folk medicine for treating infectious and stress related diseases.

Solvent–solvent fractionations of Combretum erythrophyllum (Burch.) leave extract: Studies of their antibacterial, antifungal, antioxidant and cytotoxicity potentials

Objective To evaluate the biological activities of Combretum erythrophyllum (C. erythrophyllum) leaf extracts against infectious diseases’ pathogenesis and their cytotoxicity potentials. Methods Powdered leaf material (300 g) of C. erythrophyllum was extracted (1:10 w/v) using acetone to obtain the crude extract. Liquid–liquid fractionation was performed on the crude acetone extract (30 g) using solvents of different polarity. The bioautographic method was used to detect the inhibition of bacterial and fungal growth by active compounds present in the crude and fractions. The extracts were then tested on bacterial strains: Staphylococcus aureus, Enterococcus faecalis, Escherichia coli, Pseudomonas aeruginosa; fungal strains: Candida albicans (C. albicans), Cryptococcus neoformans, and Aspergillus fumigatus, by microtitre dilution method for MIC determination. Results The extracts MIC values ranged between 0.08 and 2.50 mg/mL against the tested pathogens. Water fraction had the highest activity against bacteria strains, while the fungal assay revealed crude acetone extract and ethyl acetate fraction to be active against C. albicans (1.25 mg/mL), dichloromethane extract against C. albicans and A. fumigatus (0.16 mg/mL). Extract fractions showed a good antioxidant activity via DPPH, ABTS and hydroxyl radical scavenging assays, in the order: ethyl acetate > water > acetone > dichloromethane > hexane. The toxicity level of crude extract and fractions evaluated in Vero monkey kidney cells ranged from 34 to 223 μg/mL, while doxorubicin (IC

TLC identification of quercetin and chlorogenic acid in Ramulus mori and their antioxidant activity screening / 国际药学研究杂志

Objective To establish a thin-layer chrometograrrgy(TLC) method for the identification of quercetin and chlorogenic acid in Ramulus mori, and to screen their antioxidant activity. Methods The quercetin and chlorogenic acid were extracted by ultrasonic method with methanol as a solvent. The effect of different developed system, reagent, temperature, view methods and different silica gel plate on the TLC of quercetin and chlorogenic acid in Ramulus mori were tested to select the best TLC conditions. The antioxidant activity of quercetin and chlorogenic acid was screened with DPPH as a reagent. Results The ethyl acetate:water:formic acid:toluene(17:2:2:0.8) was used as a developing system and 1% AlCl3 as a chromogenic reagent. Quercetin and chlorogenic acid in Ramulus mori were identified under 366 nm, with blue and blue-green spots on silica gel G plate, and yellowish spots under purple background by the test of TLC-bioautography. Both were proven to have antioxidant activity. Conclusion The method is simple, accurate and reliable, and can be used for quality control of Ramulus mori.

TLC identification of quercetin and chlorogenic acid in Ramulus mori and their antioxidant activity screening / 国际药学研究杂志

Objective To establish a thin-layer chrometograrrgy(TLC)method for the identification of quercetin and chloro?genic acid in Ramulus mori,and to screen their antioxidant activity. Methods The quercetin and chlorogenic acid were extracted by ultrasonic method with methanol as a solvent. The effect of different developed system,reagent,temperature,view methods and differ?ent silica gel plate on the TLC of quercetin and chlorogenic acid in Ramulus mori were tested to select the best TLC conditions. The antioxidant activity of quercetin and chlorogenic acid was screened with DPPH as a reagent. Results The ethyl acetate∶water∶formic acid∶toluene(17∶2∶2∶0.8)was used as a developing system and 1%AlCl3 as a chromogenic reagent. Quercetin and chlorogenic acid in Ramulus mori were identified under 366 nm,with blue and blue-green spots on silica gel G plate,and yellowish spots under purple background by the test of TLC-bioautography. Both were proven to have antioxidant activity. Conclusion The method is simple,accu?rate and reliable,and can be used for quality control of Ramulus mori.

Bioautography and its scope in the field of natural product chemistry / 药物分析学报

Medicinal plants, vegetables and fruits are the sources of huge number of bioactive lead/scaffolds with therapeutic and nutraceutical importance. Bioautography is a means of target-directed isolation of active molecules on chromatogram. Organic solvents employed in chromatographic separation process can be completely removed before biological detection because these solvents cause inactivation of enzymes and/or death of living organisms. They offer a rapid and easy identification of bioactive lead/scaffolds in complex matrices of plant extracts. Bioautography is a technique to isolate hit(s)/lead(s) by employing a suitable chromatographic process followed by a biological detection system. This review critically describes the methodologies to identify antimicrobial, antioxidant, enzyme inhibitor lead/scaffolds by employing bioautography. A significant number of examples have been incorporated to authenticate the methodologies.

Partial Purification and Antimycobacterial Screening of the Ethyl Acetate Extract of Alcaligenes faecalis BW1.

Aims: The focus of this study was to evaluate the antimycobacterial activity of Alcaligenes faecalis BW1 extract and to purify it partially. Study design: Partial purification of A. faecalis BW1 extract was performed by using thin layer chromatography and active substances responsible for the biological activity were localized. Place and Duration of Study: The study was carried out at laboratory of Microbial Biotechnology, Department of Biology, Faculty of Sciences and Technical, University Sidi Mohamed Ben Abdellah, BP 2202, Road of Immouzer, Fez, Morocco, during the period from January 2011 to July 2011. Methodology: Crude extract of A. faecalis BW1 was obtained by using ethyl acetate as an organic solvent and its antimycobacterial effect was investigated by agar discs diffusion method. The extract was then fractionated by thin layer chromatography and the bioactivity was assessed with a bioautography technique followed by spots elution tests. Results: The results showed that A. faecalis BW1 produced compounds with antimycobacterial activity. All the detected spots by thin layer chromatography inhibited the growth of M. smegamtis. Conclusion: Various metabolites of A. faecalis BW1 are responsible for the sought effect or they could act synergistically to inhibit mycobacterial growth. These compounds could be used after their total purification in further work against mycobacterial infections.

Isolation and identification of anti-methicillin resistant Staphylococcus aureus compounds from Phyllanthus columnaris stem bark

Aim: Phyllanthus columnaris Müll.Arg. was found to possess anti-methicillin resistant Staphylococcus aureus (antiMRSA) activities. This study aimed at isolating, identifying and evaluating the active compounds from the stem bark of Phyllanthus columnaris Müll.Arg. against MRSA. Methodology and results: Stem bark extracts (methanol, acetone and aqueous) of Phyllanthus columnaris were subjected to anti-MRSA screening by disc diffusion method. MIC and MBC tests were carried out to compare the lowest concentration to inhibit and kill the sixteen MRSA tested among the three extracts. TLC bioautography were performed to detect the bioactive compounds. Isolation of the two active compounds was performed by means of preparative TLC. Morphological and ultra-structure alterations of the MRSA treated with bioactive compounds after 24 h were revealed by scanning and transmission electron microscopy. Both methanol and acetone extracts exhibited good anti-MRSA activity with the lowest minimum inhibitory concentration (MIC) value for both extracts were 0.78 mg/mL and the lowest minimum bactericidal concentration (MBC) were 1.56 mg/mL. Bioassay-guided chromatography by bioautography revealed two active anti-MRSA compounds from both tannin-free methanol and acetone extracts and characterized as stigmasterol and lupeol by nuclear magnetic resonance (NMR) spectral data. Scanning and transmission electron microscopy of MRSA treated with stigmasterol and lupeol showed cell wall disruption, release of cytoplasmic compounds and decreased in cellular volume. Conclusion, significance and impact of study: Results obtained herein, may suggest that the stem bark of Phyllanthus columnaris possess anti-MRSA and the two of the active compounds isolated were stigmasterol and lupeol. Their anti-MRSA effects up to the morphological and ultra-structure studies were not reported earlier

Trividha Snehapaka of Panchagavya Ghrita: A critical comparative evaluation.

Backgorund: The three stages of Snehapaka formulations namely Mridu, Madhyama and Khara Paka have been characteristically advocated for different routes of administration—Nasya, Pana/Basti and Abhyanga, respectively. Guidelines or established method for post-formulation characterization for the same is hardly available. Objective: The present communication is the comparative study of Mridu, Madhyama and Khara Paka of Panchagavya ghrita (PGG). Materials and Methods: Laboratory prepared samples of PGG following classical method were analyzed for different physicochemical, spectroscopic, chromatographic parameters, and antioxidant activity. Results: No significant difference was found among Mridu, Madhyama and Khara Paka in physicochemical parameters as well as chromatographic profiles. The ratio of absorbance at 240 and 294 nm showed steady increase from Mridu to Madhyama to Khara Paka in the ultraviolet (UV)-visible spectra of unsaponifiable matter. The high performance thin layer chromatography (HPTLC)-2,2 Diphenyl-1-picryl hydrazil (DPPH) bioautography assay revealed presence of two antioxidant compounds in low concentration in all the samples. This was further supported by estimation of total reducing power and DPPH assay. No significant difference was found among the three samples. Conclusion: Comparison of various physicochemical parameters, chromatographic profiles, and in vitro antioxidant activity determination is of little help in establishing any significant difference among the samples. However, spectrophotometric analysis of unsaponifiable matter reveals some encouraging characteristic findings which will be useful in establishing difference among the three stages of processing of PGG as well as Snehapaka in general.

Antimicrobial activity of Pavetta indica leaves.

Antimicrobial activity of the aqueous and organic solvent extracts of the leaves of Pavetta indica were tested against Bacillus subtilis, Escherichia coli and Saccharomyces cerevisiae using disc diffusion assay. Most of the leaf extracts showed bactericidal activity against B. subtilis. None of the extracts exhibited any activity against E. coli and S. cerevisiae. Minimum inhibitory concentration (MIC), minimum bactericidal concentration (MBC), thermal stability and qualitative phytochemicals studies were performed. Both MIC and MBC of the aqueous and methanol extracts were found to be between 1.95 - 7.81 mg/ml. The activity of aqueous and methanol extracts were found to be stable despite thermal treatment. Phytochemical analysis of aqueous extract revealed the presence of flavonoids, saponins and carbohydrates. Methanol extract was found to be positive for saponin and cardiac glycosides. TLC and bioautography were also done to identify the active fractions responsible for the antimicrobial activities. Results showed the presence of a number of bactericidal components. The study suggests P. indica to be a source for isolation of antibacterial compounds for human health care and use as preservatives in food processing industry.

Bioautography to assess antibacterial activity of Ottonia martiana Miq. (Piperaceae) on the human oral microbiota

Ottonia martiana Miq. (Piperaceae), planta conhecida popularmente por "anestésia" e empregada no tratamento de odontalgias devido à sua ação anestésica sobre a mucosa oral, foi investigada por meio de ensaios antibacterianos de difusão em disco de papel e de bioautografia frente a microorganismos presentes na microbiota oral humana [Streptococcus mutans (ATCC 25175), Streptococcus mitis (ATCC 49456), Streptococcus pyogenes (ATCC 19615), Streptococcus salivarius (ATCC 25975), Escherichia coli (ATCC 11229 e 25922), Pseudomonas aeruginosa (ATCC 27853), Enterobacter aerogenes (ATCC 27853). Os resultados dos bioensaios mostraram que o extrato bruto de O. martiana (32.9 mg mL-1) apresenta potencial antibacteriano frente às bactérias Gram-positivas testadas. Dentre as substâncias bioativas detectadas foram identificadas a piperovatina (Rf 0.35), piperlonguminina (Rf 0.52) e a isopiperlonguminina (Rf 0.52). A piperovatina e isopiperlonguminina foram isoladadas do extrato das raízes de O. martiana, guiadas pelo teste de bioautografia.

Ottonia martiana Miq. (Piperaceae), a plant known popularly in southern Brazil as "anestésia" and used in the treatment of odontalgia for its anesthetic action on the oral mucosa, was investigated for antibacterial activity by paper disc agar diffusion and bioautographic methods, against microorganisms present in the human oral cavity [Streptococcus mutans (ATCC 25175), Streptococcus mitis (ATCC 49456), Streptococcus pyogenes (ATCC 19615), Streptococcus salivarius (ATCC 25975), Escherichia coli (ATCC 11229 and 25922), Pseudomonas aeruginosa (ATCC 27853) and Enterobacter aerogenes (ATCC 27853).The crude extract of O. martiana (32.9 mg mL-1) had antibacterial potential against all Gram-positive bacteria tested. Analysis of the bioautograms led to the detection of bioactive substances, among which it was possible to identify piperovatine (Rf 0.35), piperlonguminine (Rf 0.52) and isopiperlonguminine (Rf 0.52). The piperovatine and isopiperlonguminine were isolated from the roots of O. martiana, guided by a bioautographic antibacterial bioassay.

Screening, isolation and purification of antibacterial agents from marine actinomycetes.

As marine environmental conditions are extremely different from terrestrial ones, it is surmised that marine actino-mycetes might produce novel bioactive compounds. Hence marine sediments, collected from the coastal areas of Gokharna and Muradeshwara of Karnataka state, were screened Seventeen isolates were obtained on starch-casein agar media by soil dilution technique. However, only six isolates namely ACT-A2, ACT-A3, ACT-A4, ACT-A5, ACT-A7 and ACT-A15 showed significant antibacterial activity against both gram-positive and gram-negative bacteria. Morphological, cultural and biochemical characterization indicated that the isolates belong to Streptomyces genus of Actinomycetes. Further studies were carried out with the most active ACT-A2. Optimization of media, temperature and pH by shake flask fermentation indicated starch-casein, 28°C and 7 to be suitable for ACT-A2. The production of antibiotics began after 24 h, reached maximum at 72 h and maintained at the same level up to 120 h. Ethyl acetate was used to extract antibacterial compounds from the culture filtrate. TLC was done on silica gel using ethyl acetate: methanol (6:4) and direct bioautography has shown the presence of two active substance, one with Rf 0.8 has more activity than the other with Rf 0.4. Further purification is done by column chromatography using a mixture of dicholoromethane and ethyl acetate. The findings from this investigation reveal that the strain ACT-A2 in order exhibited superior antimicrobial activities to other sediment isolates of actinomycetes.

Exploitation of bacteria from forest ecosystem for antimicrobial compounds.

This study deals with bacterial prospecting from forest soil with special reference to antimicrobial substances. Total of 25 morphologically different bacterial colonies were isolated from soil samples collected from Anaimalai forest and Parambikulam tiger reserve forest, Western Ghats, India. About 12 (48%) out of 25 isolates showed antibacterial activity in which strain AF1 showed inhibitory activity against more number of test pathogens. Bioactive substance from strain AF1 was produced by adopting submerged fermentation and extracted using ethyl acetate and chloroform. In disc diffusion method, ethyl acetate extract showed good antibacterial activity (9-17 mm zone of inhibition). Active fraction present in the ethyl acetate extract was determined by thin layer chromatography based bioautography. Findings of this work supported that the forest ecosystems investigated in this study will be potential place for bacterial bioprospecting.

Screening, isolation and purification of antibacterial agents from marine actinomycetes.

As marine environmental conditions are extremely different from terrestrial ones, it is surmised that marine actino-mycetes might produce novel bioactive compounds. Hence marine sediments, collected from the coastal areas of Gokharna and Muradeshwara of Karnataka state, were screened Seventeen isolates were obtained on starch-casein agar media by soil dilution technique. However, only six isolates namely ACT-A2, ACT-A3, ACT-A4, ACT-A5, ACT-A7 and ACT-A15 showed significant antibacterial activity against both gram-positive and gram-negative bacteria. Morphological, cultural and biochemical characterization indicated that the isolates belong to Streptomyces genus of Actinomycetes. Further studies were carried out with the most active ACT-A2. Optimization of media, temperature and pH by shake flask fermentation indicated starch-casein, 28°C and 7 to be suitable for ACT-A2. The production of antibiotics began after 24 h, reached maximum at 72 h and maintained at the same level up to 120 h. Ethyl acetate was used to extract antibacterial compounds from the culture filtrate. TLC was done on silica gel using ethyl acetate: methanol (6:4) and direct bioautography has shown the presence of two active substance, one with Rf 0.8 has more activity than the other with Rf 0.4. Further purification is done by column chromatography using a mixture of dicholoromethane and ethyl acetate. The findings from this investigation reveal that the strain ACT-A2 in order exhibited superior antimicrobial activities to other sediment isolates of actinomycetes.

Evaluation of antimicrobial and phytochemical screening of Fennel, Juniper and Kalonji essential oils against multi drug resistant clinical isolates

Objective: The inhibitory effects of essential oils including fennel, juniper and kalonji from Foeniculum Vulgare, Juniperus Osteosperma and Nigella Sativa on multi drug resistant clinical isolates were investigated. All the oils have been evaluated for phytochemical constituents, antibacterial activity and TLC bioautography assay. Methods: Preliminary phytochemical analysis was performed. The antibacterial potential of essential oils from fennel, juniper and kalonji fennel, juniper and kalonji was evaluated by agar well diffusion method against multi drug resistant clinical isolates. The antibacterial effect was investigated using the TLC-bioautographic method. Results: Preliminary phytochemical analysis demonstrated the presence of most of the phytochemicals including saponins, cardiac glycosides, steroids, terpenoids, flavonoids and tannins. Antibacterial activity of essential oils was assessed on eight multi-drug resistant (MDR) clinical isolates from both Gram-positive and Gram-negative bacteria and two standard strains. All the oils tested showed significant to moderate antibacterial activity toward all tested strains except Acinetobacter sp and Staphylococcus aureus MRSA. The maximum zone of inhibition was found to be 25依0.12 mm for juniper oil followed by 21依0.085 mm for kalonji oil againstStaphylococcus aureus 2. Thin layer chromatography and bioautography assay demonstrated well-defined growth inhibition zones against Staphylococcus aureus 2 and E. coli for juniper essential oil in correspondence with tannins observed at Rf values of 0.07 and 0.57. Conclusions: Based on the present study, the essential oils from juniper and kalonji possess antibacterial activity against several multi drug resistant pathogenic bacteria and thus can be used as a base for the development of new potent drugs and phytomedicine.

Screening of Amazonian plants from the Adolpho Ducke forest reserve, Manaus, state of Amazonas, Brazil, for antimicrobial activity

Tropical forests are species-rich reserves for the discovery and development of antimicrobial drugs. The aim of this work is to investigate the in vitro antimicrobial potential of Amazon plants found within the National Institute on Amazon Research's Adolpho Ducke forest reserve, located in Manaus, state of Amazonas, Brazil. 75 methanol, chloroform and water extracts representing 12 plant species were tested for antimicrobial activity towards strains of Mycobacterium smegmatis, Escherichia coli, Streptococcus sanguis, Streptococcus oralis, Staphylococcus aureus and Candida albicans using the gel-diffusion method. Active extracts were further evaluated to establish minimum inhibitory concentrations (MIC) and antimicrobial profiles using bioautography on normal-phase thin-layer chromatography plates. Diclinanona calycina presented extracts with good antimicrobial activity and S. oralis and M. smegmatis were the most sensitive bacteria. D. calycina and Lacmellea gracilis presented extracts with the lowest MIC (48.8 µg/ml). D. calycina methanol and chloroform leaf extracts presented the best overall antimicrobial activity. All test organisms were sensitive to D. calycina branch chloroform extract in the bioautography assay. This is the first evaluation of the biological activity of these plant species and significant in vitro antimicrobial activity was detected in extracts and components from two species, D. calycina and L. gracilis.