Bioautography Assay Research Articles

Isolation and Inhibitory Activity Testing of Alpha-Glucosidase Enzyme from Endophytic Bacteria in Kumis Kucing (Orthosiphon aristatus (Blume) Miq.) Leaves

Orthosiphon aristatus (Blume) Miq., commonly known as kumis kucing, is frequently used in traditional Indonesian medicine to treat diabetes, hypertension, rheumatism, gout, and as a diuretic. However, large-scale production of medicinal products from natural sources requires significant raw material and land, creating a need for alternative methods. This study explores the potential of isolating endophytic bacteria from kumis kucing leaves to serve as α-glucosidase enzyme inhibitors, offering a more sustainable approach. Eight bacterial endophytes were isolated and tested using bioautography assays and IC50 measurements to assess their inhibitory activity. Four isolates—KK 1, KK 3, KK 4, and KK 5—demonstrated significant α-glucosidase inhibition, with IC50 values of 41.35 µg/mL, 57.04 µg/mL, 56.70 µg/mL, and 164.16 µg/mL, respectively. Molecular testing identified KK 1 as Priestia aryabhattai, while KK 3 and KK 5 were identified as Priestia megaterium. These findings suggest that endophytic bacteria isolated from Orthosiphon aristatus leaves have the potential to be developed as natural sources of α-glucosidase inhibitors, which are beneficial for diabetes management.

Ethnobotanical survey and anti-candidal activity of plant species used for oral candidiasis

Background: Oral candidiasis is caused by Candida albicans, which is most prevalent in immunocompromised patients.Aim: The study aimed to investigate the antifungal activity of plant species used for oral candidiasis against C. albicans.Setting: The study was conducted in Aganang Local Municipality, Capricorn District, Limpopo province, South Africa.Methods: A survey was conducted using a semi-structured questionnaire supplemented with guided field walks with traditional health practitioners to gather information on medicinal plants used to treat oral candidiasis. Nine plant species (Artemisia afra Jacq. ex Willd., Blepharis subvolubilis subsp. subvolubilis C.B. Clarke, Enicostemma axillare [Lam.], Helichrysum caespititium [DC.] Harv., Solanum incanum L., Waltheria indica L., Ximenia caffra Sond. var. caffra, Ximenia caffra Sond. var. natalensis and Ziziphus mucronata Willd.) were investigated for antifungal activity. The plant material were extracted with solvents of varying polarities: acetone, dichloromethane, ethyl acetate, ethanol, hexane, methanol, and water. The Micro-dilution and bioautography assays were used to determine the antifungal activity of the plant extracts.Results: Leaf extracts of A. afra and S. incanum were more active against C. albicans with MIC values of 0.02 mg/mL. Bioautography assay demonstrated active compounds in S. incanum, W. indica and X. caffra var. caffra extracts developed in Benzene: Ethanol: Ammonia hydroxide (BEA).Conclusion: An ethnobotanical survey is a worthy starting point in selecting potential plant species for ethnopharmacological studies.Contribution: The effectiveness of oral administrations of the medicinal plants was confirmed by the excellent antifungal activity of the aqueous extracts.

Thin layer chromatography assay to detect laccase inhibitors

Thin-layer chromatography (TLC) hyphenated to bioassays is a modern tool used for discovery of biologically active compounds from complex mixtures. The first bioautographic assay for detecting laccase inhibitors on a TLC plate was developed in this study. The on-plate reaction of laccase with colourless ABTS that renders the blue ABTS∙+ radical was optimised. Combination of the enzymatic TLC-assay with a control TLC-assay, wherein ABTS∙+ radical is chemically generated and then applied on the TLC, allowed to differentiate between the pure laccase inhibitor sodium azide and radical scavengers such as gallic and kojic acids. The limit of detection and quantification for the method were 54.9 and 166 ng of sodium azide respectively. The methodology was applied successfully to a recently discovered laccase inhibitor chemotype: hydrazones. A model hydrazone was compared with several hydrazones synthesized for this study. For the first time, laccase inhibitors separated on a TLC plate can be detected individually.

Preliminary phytochemical and biological activity screening of Sargassum lapazeanum

Algae are exposed to substantial stress. In response, these organisms have developed efficient defense systems, such as protective secondary metabolite synthesis, making algae a primary source of bioactive compounds with a wide spectrum of biological activities. Thus, algae show potential for use in treatments of thrombotic, infectious, and chronic degenerative diseases. Therefore, the objective of this study was to evaluate the phytochemical compounds and pharmacological activity of an extract obtained from Sargassum lapazeanum. Algae were collected in the intertidal zone of Tarabillas beach (Bahía de La Paz, BCS). The biological activities of an ethanolic extract and its fractions were evaluated using chromatographic techniques. In addition, a bioautographic assay of hemolytic activity was conducted, and phytochemical profiles and acute toxicity in Artemia franciscana were evaluated. The relationships among the main extract components were also determined. The ethanolic extract exhibited significant antioxidant and hemolytic activity, which was mainly attributed to its content of anthrones, anthraquinones, and unsaturated triterpenes. Its toxicological activity reached an LC50 value of 225.1 μg mL-1, which was mainly attributed to alkaloids, flavonoids, anthrones, and saponins. The results suggest that Sargassum lapazeanum, which is endemic to the Gulf of California, has great pharmacological potential with biomedical applications.

Benzyl-isoquinoline alkaloids rich extract of Coptis teeta Wall., exhibit potential efficacy in calcium-oxalate and uric-acid linked metabolic disorders

Coptis teeta Wall., an endangered but valuable medicinal species having various folklore uses in Indian and Chinese Traditional system of medicine. Its distribution is restricted to India, China and Tibet. In India, C. teeta is traditionally used in joint disorders, urinary infections and inflammatory diseases, however the scientific validation is missing. Thus, the present study aims to validate the anti-lithiatic and anti-gout activity of C. teeta rhizome extract (CTME) through in-vitro biological assays. The metabolic fingerprinting of CTME through reverse phase-high performance liquid chromatography-photodiode array (RP-HPLC-PDA) showed the presence of five benzyl-isoquinoline alkaloids, namely berberine (2.59%), coptisine (0.746%) jatrorrhizine (0.133%), palmatine (0.03%) and tetrahydropalmatine (0.003%). The anti-gout potency analysed via in-vitro xanthine oxidase (XOD) inhibition assay, followed by HPTLC (High performance thin layer chromatography) mediated bio-autographic inhibition of XOD signifies that CTME exhibit strong inhibition of XOD (IC50: 3.014 μg/ml), insignificantly different (p > 0.05) from allopurinol (IC50: 2.47 μg/ml). The XOD bioautographic assay advocates that the efficacy is primarily due to berberine and coptisine alkaloids. The CTME has significant anti-lithiatic activity, and thereby limiting the progression of crystal nidus formation, mediated via inhibition of calcium oxalate crystals nucleation and aggregation. Additionally, the extract also exhibits potential effect on inhibition of oxidative stress associated inflammation, which plays crucial role in alleviating urolithiasis and gouty conditions. Validating the traditional claims of C. teeta will not only confirm its medicinal benefits for targeted pathological conditions but also enhance its industrial demand.

Antimicrobial Activity of Fungal Endophytes Associated with Peperomia argyreia (Piperaceae)

The endophytic fungal biodiversity of unique plants like Peperomia argyreia (Miq.) É. Morren (Piperaceae) has antimicrobial properties and can be employed for infection treatment. Fungal isolates were obtained from appropriately treated plant tissues cultured in solid media, characterized by morphology, and identified by molecular biology using ITS and NL primers. The antimicrobial properties of fungal extracts were analyzed by combining microdilution and bioautographic assays complemented with metabolic profiling by automated thin-layer chromatography and 1H NMR techniques. Thirty-one filamentous fungi were isolated and characterized by ITS and/or D1/D2 region amplification of rDNA, identified as Thermothielavioides, Trichoderma, Cyphellophora, Cladosporium, Arcopilus, Plectosphaerella; Chaetomium, Sporothrix, Alboefibula, and Penicillium. Thermothielavioides spp. inhibited Staphylococcus aureus ATCC 25923; moreover, Penicillium westlingii P4 showed inhibitory activity on Ascochyta rabiei AR2. The bioactivity-guided fractionation of the EtOAc extract (MIC = 62.5 μg/mL) of P. westlingii P4 allowed the purification of citrinin as the main inhibitory compound (MIC = 62.5 μg/mL). Peperomia argyreia harbors a rich and diverse endophytic community able to produce bioactive molecules. Citrinin, with a minor influence of volatile compounds biosynthesized by P. westlingii P4, was responsible for the inhibition of A. rabiei AR2.

Molecular modelling and anticholinesterase activity of the essential oil from three chemotypes of Lippia alba (Mill.) N.E.Br. ex Britton & P. Wilson (Verbenaceae)

Lippia alba (Mill.) N.E. Brown (Verbenaceae), popularly known as “erva cidreira”, is one of the most used plants in Brazilian folk medicine. The species has several chemotypes and its volatile constituents have already been characterized, and present different chemical markers with known pharmacological properties, such as analgesic, sedative and antifungal properties. The objective of this study was to evaluate the anticholinesterase activity (AChE) of the essential oil of three chemotypes of Lippia alba and, by using molecular anchoring, determine the best receptor-ligand interaction energies of the main constituents present in the samples of oil. The essential oils were obtained via hydrodistillation (LA1 and LA2) and steam drag (LA3), and their volatile constituents determined using GC-MS. For the determination of anticholinesterase activity, direct bioautography and colorimetry assays based on Ellman's method were used. Molecular docking was performed using a multiple solution genetic algorithm and Merck molecular force field 94 (MMFF94) as the scoring function. In the main constituents of the oil samples, three chemotypes were identified for L. alba: LA1 is rich in citral, LA2 is rich in carvone and LA3 is rich in linalool. All L. alba chemotypes showed AChE enzyme inhibition with an IC50 of 3.57 μg/mL (LA1), 0.1 μg/mL (LA2) and 4.34 μg/mL (LA3). The molecular docking study complemented the results of the experiment and demonstrated significant interactions between the main constituents of the oils and the amino acid residues of the AChE enzyme. Irrespective of the chemotype, Lippia alba presents biotechnological potential for the discovery of anticholinesterase substances, with the chemotype LA2 (rich in carvone) being the most active.

Mining Biosynthetic Gene Clusters of Pseudomonas vancouverensis Utilizing Whole Genome Sequencing.

Natural product (NP)-based pesticides have emerged as a compelling alternative to traditional chemical fungicides, attracting substantial attention within the agrochemical industry as the world is pushing toward sustainable and environmentally friendly approaches to safeguard crops. Microbes, both bacteria and fungi, are a huge source of diverse secondary metabolites with versatile applications across pharmaceuticals, agriculture, and the food industry. Microbial genome mining has been accelerated for pesticide/drug discovery and development in recent years, driven by advancements in genome sequencing, bioinformatics, metabolomics/metabologenomics, and synthetic biology. Here, we isolated and identified Pseudomonas vancouverensis that had shown antifungal activities against crop fungal pathogens Colletotrichum fragariae, Botrytis cinerea, and Phomopsis obscurans in a dual-plate culture and bioautography assay. Further, we sequenced the whole bacterial genome and mined the genome of this bacterium to identify secondary metabolite biosynthetic gene clusters (BGCs) using antiSMASH 7.0, PRISM 4, and BAGEL 4. An in-silico analysis suggests that P. vancouverensis possesses a rich repertoire of BGCs with the potential to produce diverse and novel NPs, including non-ribosomal peptides (NRPs), polyketides (PKs), acyl homoserine lactone, cyclodipeptide, bacteriocins, and ribosomally synthesized and post-transcriptionally modified peptides (RiPPs). Bovienimide-A, an NRP, and putidacin L1, a lectin-like bacteriocin, were among the previously known predicted metabolites produced by this bacterium, suggesting that the NPs produced by this bacterium could have biological activities and be novel as well. Future studies on the antifungal activity of these compounds will elucidate the full biotechnological potential of P. vancouverensis.

Antarctic bryophyte Sanionia uncinata (HEDW.) Loeske, Amblystegiaceae, antimicrobial, antioxidant, cytotoxic, and acetylcholinesterase activities.

Sanionia uncinata, or Sickle-leaved-Hook-moss, is a cosmopolitan pleurocarpous moss composing the Antarctic Peninsulae biodiversity, primordially forming dense mats over rocks. The species was collected in 24 different spots located at King George Island and was processed to obtain 24 ethanolic extracts (ADS#) by a serial-24h-maceration, which were prospected for antimicrobial, cytotoxic, antioxidant, and acetylcholinesterase (AChE) inhibition activities by using in vitro tests. Alien material was removed from the non-sterilized plant samples before being submitted for extraction. It was observed that extracts collected in different spots showed different biological activities. Extracts ADS04(10.66±0,17mm) and ADS14(11.37±0,11mm) were active against Staphylococcus aureus, according to the diffusion in bioautography assay. They showed significant antioxidant activity and inhibited AChE; the cytotoxicity observed to the human breast cancer cells MCF-7 and MDA-MB-231 were higher than in normal cell line MCF-10A. ADS04 was 7.62 times more cytotoxic to MCF-7, and ADS14 was 2.03 times more cytotoxic to MDA-MB-231 than to MCF-10A. The extracts showed similar cytotoxicity between PC-3, a human prostate cancer cell line, and MCF-10A. Sanionia uncinata extracts are a vital potential source of biologically active compounds, particularly ADS04 and ADS14, including further prospection on eventual bryophyte's endophytic fungi.

Antifungal activity and toxicity of bioactive compounds isolated from the leaf of Ximenia caffra Sond. var. natalensis

Background: Based on high frequency index, Ximenia caffra Sond. var. natalensis was selected for further phytochemical investigation and biological assays.Aim: The study aimed to isolate the active antifungal compounds from the leaves of X. caffra var. natalensis.Setting: The ethnobotanical study was conducted in Aganang Local Municipality, Capricorn District.Methods: Acetone extract was partitioned five times with hexane, chloroform, ethyl acetate, butanol and water, respectively. Fractions were screened for antifungal activity against Candida albicans using the microplate method and bioautography assays. The structures of isolated compounds were identified using nuclear magnetic resonance (NMR) and mass spectrometry. Cytotoxicity of isolated compounds was determined using 3-(4,5-dimethylthiazol)-2,5-diphenyl tetrazolium bromide (MTT) assay.Results: Bioassay-guided fractionation of the ethyl acetate fraction led to the isolation of four compounds, out of which only two were identified. Compound 1 was identified as epigallocatechin gallate, and Compound 3 was confirmed as kaempferol-3-O-rhamnoside. Epigallocatechin gallate exhibited moderate antifungal activity with minimum inhibitory concentration (MIC) of 0.5mg/mL and less toxic to the cells with LC50 = 32.32 µg/mL.Conclusion: The antifungal activity and cytotoxicity of isolated compounds validate the use of X. caffra Sond. var. natalensis in combating oral candidiasis.Contribution: The results have shown the potential bioactivity of X. caffra Sond. var. natalensis in the treatment of oral candidiasis.

Antifungal and Phytotoxic Activities of Isolated Compounds from Helietta parvifolia Stems

The identification of natural and environmentally friendly pesticides is a key area of interest for the agrochemical industry, with many potentially active compounds being sourced from numerous plant species. In this study, we report the bioassay-guided isolation and identification of phytotoxic and antifungal compounds from the ethyl acetate extract of Helietta parvifolia stems. We identified eight compounds, consisting of two coumarins and six alkaloids. Among these, a new alkaloid, 2-hydroxy-3,6,7-trimethoxyquinoline-4-carbaldehyde (6), was elucidated, along with seven known compounds. The phytotoxicity of purified compounds was evaluated, and chalepin (4) was active against Agrostis stolonifera at 1 mM with 50% inhibition of seed germination and it reduced Lemna pausicotata (duckweed) growth by 50% (IC50) at 168 μM. Additionally, we evaluated the antifungal activity against the fungal plant pathogen Colletotrichum fragariae using a thin-layer chromatography bioautography assay, which revealed that three isolated furoquinoline alkaloids (flindersiamine (3), kokusagenine (7), and maculine (8)) among the isolated compounds had the strongest inhibitory effects on the growth of C. fragariae at all tested concentrations. Our results indicate that these active natural compounds, i.e., (3), (4), (7), and (8), could be scaffolds for the production of more active pesticides with better physicochemical properties.

Screening Amazon rainforest plant extracts for antimicrobial activity: a 15-year commitment to the Brazilian biodiversity

IntroductionThe need for new tools to treat infections is constantly growing due to the possibilities of emerging diseases related to environmental changes, climatic catastrophes, microorganism resistance, and human and animal aging, leading to an evident unbalance in the planet’s health. Brazil contains the most significant portion of world biodiversity, a potential source of new antimicrobial natural products. Nonetheless, its environment, particularly its forests, and rainforests, is under threat, meaning that rapidly conducted, comprehensive research into the potential of antimicrobial activity to address this threat is urgently needed.MethodsIn this study, plants from the Amazon rainforest and the Atlantic forests were collected and tested against several pathogenic microbes relevant to humans, animals, and the environment, and subjected to large-scale susceptibility assays, bioautography, and Artemia salina toxicity assays. From the plants, 2,280 organic and aqueous extracts were obtained from different organs, namely leaves, barks, flowers, fruits, and seeds, and subjected to a large-scale susceptibility screening assay against Staphylococcus aureus, Staphylococcus epidermidis, Enterococcus faecalis, Streptococcus mutans, Streptococcus sanguinis, Escherichia coli, Pseudomonas aeruginosa, Candida albicans, Malassezia pachydermatis, Malassezia furfur, and Listeria monocytogenes. Results and discussionThe selected extracts were subjected to antimicrobial susceptibility tests to determine their inhibition zone diameters and minimum bactericidal concentrations, to bioautography, and to an Artemia salina toxicity assay, which resulted in 154 active extracts. Moreover, 111 out of 154 extracts were ranked based on scores established by the p-values and the mean rank differences in each set of test results. The final ranking identified which extracts should be studied in further phytochemical research using thin-layer chromatography techniques as a priority. The extracts obtained from plants belonging to Combretaceae, Connaraceae, Convolvulaceae, Fabaceae, Malpighiaceae, Moraceae, Piperaceae, Polygonaceae, and Salicaceae were selected as the most promising ones and used to support the identification of plant-based antimicrobial active compounds from the immense biodiversity of Brazilian forests.

Determining the optimum mixture of three essential oils for potato sprout suppression at room temperature storage.

As a staple crop, potatoes (Solanum tuberosum) play an important role in meeting daily caloric needs. To ensure adequate supplies for year-round consumption, potato quality must be maintained throughout lengthy storage periods. Towards this end, potato sprouting during storage must be minimized. Due to changing regulations regarding chemical means of potato sprout suppression, increased focus has turned to alternative products including essential oils (EO) as sprout suppressants in recent years. The complex composition of various EOs promises numerous options for sprout suppression. Furthermore, blends of several EOs may achieve enhanced sprout suppressant properties if synergistic interactions are present. We evaluated Syzygium aromaticum, Artemisia herba-alba, and Laurus nobilis EOs and blends thereof as sprout suppressants in potato cultivar Ranger Russet stored at room temperature and also tested for their antifungal activity against Colletotrichum fragariae, a causal organism of anthracnose disease in strawberries including other vegetables and fruits. A. herba-alba EO was an effective sprout suppressant when used alone and suppressed sprouting over the 90-day storage period. Interactions between A. herba-alba and S. aromaticum affected sprout length whereas interactions between A. herba-alba and L. nobilis EOs affected sprout number. An optimum blend of 50% - 82.31% A. herba-alba, 17.69% - 50% L. nobilis, and 0% - 1.01% S. aromaticum EOs could more effectively minimize tuber sprout length and number than any of the three whole EOs used alone. Among these three EOs, only S. aromaticum EO showed antifungal activity against C. fragariae in bioautography assay. These results exhibit the potential of EOs blends as a novel tactic in potato sprout suppression as well as potential natural product-based fungicides in managing C. fragariae.

Antibacterial effect of essential oils and their components against Xanthomonas arboricola pv. pruni revealed by microdilution and direct bioautographic assays.

Bacterial spot of stone fruits caused by Xanthomonas arboricola pv. pruni (Xap) is one of the most significant diseases of several Prunus species. Disease outbreaks can result in severe economic losses while the control options are limited. Antibacterial efficacy of essential oils (EOs) of thyme, cinnamon, clove, rosemary, tea tree, eucalyptus, lemon grass, citronella grass, and lemon balm was assessed against two Hungarian Xap isolates. The minimal inhibitory concentration (MIC) was determined by broth microdilution assay and for the identification of active EOs' components a newly introduced high-performance thin-layer chromatography (HPTLC)-Xap (direct bioautography) method combined with solid-phase microextraction-gas chromatography/mass spectrometry (SPME-GC/MS) was applied. All EOs inhibited both bacterium isolates, but cinnamon proved to be the most effective EO with MIC values of 31.25 µg/mL and 62.5 µg/mL, respectively. Compounds in the antibacterial HPTLC zones were identified as thymol in thyme, trans-cinnamaldehyde in cinnamon, eugenol in clove, borneol in rosemary, terpinen-4-ol in tea tree, citral (neral and geranial) in lemon grass and lemon balm, and citronellal and nerol in citronella grass. Regarding active compounds, thymol had the highest efficiency with a MIC value of 50 µg/mL. Antibacterial effects of EOs have already been proven for several Xanthomonas species, but to our knowledge, the studied EOs, except for lemon grass and eucalyptus, were tested for the first time against Xap. Furthermore, in case of Xap, this is the first report demonstrating that direct bioautography is a fast and suitable method for screening anti-Xap components of complex matrices, like EOs.

Flowering phenophases influence the antibacterial and anti-biofilm effects of Thymus vulgaris L. essential oil

BackgroundEssential oils are becoming increasingly popular in medicinal applications because of their antimicrobial effect. Thymus vulgaris L. (Lamiaceae) is a well-known and widely cultivated medicinal plant, which is used as a remedy for cold, cough and gastrointestinal symptoms. Essential oil content of thyme is responsible for its antimicrobial activity, however, it has been reported that the chemical composition of essential oils influences its biological activity. In order to explore flowering phenophases influence on the chemical composition of thyme essential oil and its antibacterial and anti-biofilm activity, plant materials were collected at the beginning of flowering, in full bloom and at the end of flowering periods in 2019.MethodsEssential oils from fresh and dried plant materials were distilled and analyzed with gas chromatography-mass spectrometry (GC-MS) and gas chromatography-flame ionization detection (GC-FID). The antibacterial activity was performed by broth microdilution and thin layer chromatography-direct bioautography (TLC-DB) assays and the anti-biofilm effect by crystal violet assay, respectively. Scanning electron microscopy was applied to illustrate the cellular changes of bacterial cells after essential oil treatment.ResultsThymol (52.33–62.46%) was the main component in the thyme essential oils. Thyme oil distilled from fresh plant material and collected at the beginning of flowering period exerted the highest antibacterial and anti-biofilm activity against Haemophilus influenzae, H. parainfluenzae and Pseudomonas aeruginosa.ConclusionThe different flowering periods of Thymus vulgaris influence the antibacterial and anti-biofilm activity of its essential oils, therefore, the collection time has to be taken into consideration and not only the full bloom, but the beginning of flowering period may provide biological active thyme essential oil.

Evaluation of hydromethanolic extract and aglycone fraction of Actiniopteris semiflabellata for antioxidant activity

Actiniopteris semiflabellata Pic-Ser grows in Yemen and is used by the local population for wounds and burns. The goal of the current study was to assess the Actiniopteris semiflabellata's chemical composition and antioxidant activity. Phenols, flavonoids, triterpenes and sterols were identified in the hydromethanolic extract and in the aglycone fraction, while carbohydrates were only found in the hydromethanolic extract. In the hydromethanolic extract and the aglycone fraction, the total phenolic content was evaluated as 17.73 mg/g and 7.22 mg/g respectively, while the total flavonoid content was 11.21 mg/g and 4.13 respectively. Studied extract and fraction exhibited antioxidant activity by TLC bioautography and DPPH radical scavenging activity assay. The IC50 values for the hydromethanolic extract and the aglycone fraction were 79.64 and 148.60 µg/ml respectively. These results confirm the traditional use and are useful for further study of this plant, in addition, it may be a good source of natural antioxidants.

Tamarindus indica Extract as a Promising Antimicrobial and Antivirulence Therapy

The worldwide crises from multi-drug-resistant (MDR) bacterial infections are pushing us to search for new alternative therapies. The renewed interest in medicinal plants has gained the attention of our research group. Tamarindus indica L. (T. indica) is one of the traditional medicines used for a wide range of diseases. Therefore, we evaluated the antimicrobial activities of ethanolic extract of T. indica. The inhibitions zones, minimum inhibitory concentration (MIC), minimum bactericidal concentration (MBC), and fractional inhibitor concentration indices (FICI) against Gram+ve and -ve pathogens were detected. The bioactive compounds from T. indica extract were identified by mass spectroscopy, thin-layer chromatography, and bio-autographic assay. We performed scanning electron microscopy (SEM) and molecular docking studies to confirm possible mechanisms of actions and antivirulence activities, respectively. We found more promising antimicrobial activities against MDR pathogens with MIC and MBC values for Staphylococcus aureus (S. aureus) and Pseudomonas aeruginosa (P. aeruginosa), i.e., (0.78, 3.12 mg/mL) and (1.56, 3.12 mg/mL), respectively. The antimicrobial activities of this extract were attributed to its capability to impair cell membrane permeability, inducing bacterial cell lysis, which was confirmed by the morphological changes observed under SEM. The synergistic interactions between this extract and commonly used antibiotics were confirmed (FICI values < 0.5). The bioactive compounds of this extract were bis (2-ethylhexyl)phthalate, phenol, 2,4-bis(1,1-dimethylethyl), 1,2-benzenedicarboxylic acid, and bis(8-methylnonyl) ester. Additionally, this extract showed antivirulence activities, especially against the S. aureus protease and P. aeruginosa elastase. In conclusion, we hope that pharmaceutical companies can utilize our findings to produce a new formulation of T. indica ethanolic extract with other antibiotics.

Antimicrobial properties and biotransforming ability of fungal endophytes from Ficus carica L. (Moraceae)

ABSTRACT The endophytic fungal community associated with leaves of Ficus carica L. (Moraceae) from Argentina was investigated. Fifteen fungal isolates were isolated and identified by molecular methods into the genera Alternaria, Cladosporium, Curvularia, Diaporthe, Epicoccum, Myrothecium, Neofusicoccum, Nigrospora, Preussia and Ustilago. Cladosporium cladosporioides and Curvularia lunata were the most frequently isolated species. The fungal metabolic profiles were obtained by automated TLC and NMR and analysed by PC Analysis. Antifungal and antibacterial activity was assessed by bioautographic assays. In addition, the biotransforming ability of the fungal isolates was tested on F. carica extracts. Five isolates (33.3%) exhibited inhibitory activity against at least one of the microorganisms tested. Most of the fungal endophytes were able to metabolise the flavonoid rutin 1, and the coumarin psoralen 3 present in F. carica extracts. Further investigations of the psoralen biotransforming ability performed by the selected endophyte Alternaria alternata F8 showed the accumulation of the 6,7-furan-hydrocoumaric acid derivative 4 as the main biotransformation product. Our results corroborate that F. carica can live symbiotically with rich and diverse endophytic communities adding insights about their ecological interactions.

Ethnobotanical Survey, Physiochemical Composition and Preliminary Cytotoxic Evaluation of some Medicinal Plants with Anticancer Potential from Certain Areas in South-West Nigeria

Aims: Medicinal plants used by traditional medical practitioners (TMP) to treat cancers are considered safe when used alone or combined with conventional therapy to ensure their effectiveness and eliminate the toxic effects of orthodox medicines. Using cytotoxic and antioxidant studies, the study attempted to assess some of the commonly used medicinal plants used to cure cancer among Yoruba people in Ogun, Oyo, Osun, and Lagos (South-West, Nigeria).&#x0D; Study Design: Samples of commonly utilized anticancer plants obtained from the chosen areas using physical and virtual oral seminars were studied for physiochemical composition and a possible antioxidant and cytotoxic potential to validate the basis for the use of the selected anticancer plants.&#x0D; Methodology: Online academic literature searches were done on the cited plants to identify the already-exploited anticancer plants. The ethanolic extracts of the plant were examined for the presence of bioactive components and their total flavonoid content, with focusing on quercetin detection using thin layer bioautography (TLB) and brine shrimp lethality assay (BSLA) for cytotoxicity. In comparison to quercetin and ascorbic acid, the scavenging of superoxide radical (SOR), hydrogen peroxide, and 2, 2-Diphenyl-2-picrylhydrazyl (DPPH) radical activity by a model (most biologically active) of the anticancer plant was also evaluated.&#x0D; Results: There were only twelve anticancer species that were not used in related studies: Lannea egregia, Ficus exasperate, Croton zambesicus, Tetrapleurai tetraptera, Terminalia catappa, Zanthoxylum zanthoxyloides, Plumbago zelanica, Hilleria latifolia, Bryophyllum pinntum, Chromolena odorata, Brysocarpus coccineus and Spondias mombin. The anticancer plants contained bioactive and mineral substances like saponins, protein, lipids, magnesium, calcium, iron, zinc, and a decreased Na/K concentration. The plants had a fair amount of flavonoids and variable levels of cytotoxicity. L. egeregia was regarded as the prototype of the anticancer species due to its profound flavonoid concentration (85.40 µg/mL) and cytotoxicity (9.46 µg/mL) compared to other extracts. The TLB also demonstrated the presence of quercetin, with a dose-dependent antioxidant property. The anticancer model's overall antioxidant activity (34.72 µg/mL) was slightly lower than quercetin (30.44 µg/mL) but higher than ascorbic acid (41.68 µg/mL).&#x0D; Conclusion: The results support the traditional use of anticancer species as nutritional and dietary supplements, whose bioactive compounds are relevant in managing cancer patients. The plant’s bioactive principles need to be characterized in future research.

A Comparative Study on Chemical Compositions and Biological Activities of Four Amazonian Ecuador Essential Oils: Curcuma longa L. (Zingiberaceae), Cymbopogon citratus (DC.) Stapf, (Poaceae), Ocimum campechianum Mill. (Lamiaceae), and Zingiber officinale Roscoe (Zingiberaceae).

Essential oils (EOs) and their vapour phase of Curcuma longa (Zingiberaceae), Cymbopogon citratus (Poaceae), Ocimum campechianum (Lamiaceae), and Zingiber officinale (Zingiberaceae) of cultivated plants grown in an Amazonian Ecuador area were chemically characterised by Gas Chromatography-Flame Ionization Detector (GC-FID), Gas Chromatography-Mass Spectrometry (GC-MS), and Head Space-Gas Chromatograph-Flame Ionization Detector-Mass Spectrometry (HS-GC-FID-MS).figure The EOs analyses led to the identification of 25 compounds for C. longa (99.46% of the total; ar-turmerone: 23.35%), 18 compounds for C. citratus (99.59% of the total; geraniol: 39.43%), 19 compounds for O. campechianum (96.24% of the total; eugenol: 50.97%), and 28 for Z. officinale (98.04% of the total; α-Zingiberene: 15.45%). The Head Space fractions (HS) revealed C. longa mainly characterised by limonene and 1,8-cineole (37.35%) and α-phellandrene (32.33%); Z. officinale and C. citratus showed camphene (50.39%) and cis-Isocitral (15.27%) as the most abundant compounds, respectively. O. campechianum EO revealed a higher amount of sesquiterpenes (10.08%), mainly characterised by E-caryophyllene (4.95%), but monoterpene fraction remained the most abundant (89.94%). The EOs were tested for antioxidant, antimicrobial, and mutagen-protective properties and compared to the Thymus vulgaris EO as a positive reference. O. campechianum EO was the most effective in all the bioactivities checked. Similar results emerged from assaying the bioactivity of the vapour phase of O. campechianum EO. The antioxidant and antimicrobial activity evaluation of O. campechianum EO were repeated through HP-TLC bioautography assay, pointing out eugenol as the lead compound for bioactivity. The mutagen-protective evaluation checked through Ames's test properly modified evidenced a better capacity of O. campechianum EO compared with the other EOs, reducing the induced mutagenicity at 0.1 mg/plate. However, even with differences in efficacy, the overall results suggest important perspectives for the functional use of the four studied EOs.