SACCHAROMONOSPORA OCEANI VJDS‑3, A POTENT ACTINOBACTERIAL STRAIN FROM MANGROVE ECOSYSTEM

Objective: The aim of the present study was to isolate, identify, and analyze the phylogenetic characteristics of a rare actinobacterial strain VJDS‑3with antagonistic activities isolated from Mangrove ecosystems of Nizampatnam, Guntur District, Andhra Pradesh, India.Methods: Soil samples collected were pre‑treated with calcium carbonate and used for isolation of potent actinobacterial strain designated as VJDS‑3.Identification of the strain was carried out by studying the micro‑morphological, cultural, biochemical, and physiological methods. The phylogeneticstudy of the strain was carried out by employing 16S rDNA sequence‑based analysis. The phylogenetic tree was constructed using the MolecularEvolutionary Genetic Analysis software version 6.Results: The potent actinobacterial strain was identified as Saccharomonospora oceani VJDS‑3, and the bioactive metabolites produced by the straininhibited Gram‑positive bacteria (Bacillus megaterium, Bacillus subtilis), Gram‑negative bacteria (Xanthomonas campestris, Pseudomonas aeruginosa,Proteus vulgaris, and Escherichia coli), and fungi (Aspergillus niger, Botrytis cinerea, Fusarium solani, Fusarium Oxysporum, and Candida albicans).Conclusion: The results of the experiment showed that the crude ethyl acetate extract of S. oceani VJDS‑3 showed significant antimicrobial potential,and hence it can be used for isolation of compounds with pharmaceutical importance.Keywords: Mangrove ecosystems, Phylogenetic study, Saccharomonospora oceani VJDS‑3, Bioactive compounds

Production of pullulan using jaggery as substrate by Aureobasidium pullulans MTCC 2195

Shake-flask fermentation, under batch cultivation, was investigated for the production of fungal exopolysaccharide, pullulan using jaggery (a traditional concentrated sugar cane juice) as a carbon substrate by Aureobasidium pullulans MTCC 2195. Change in the initial pH (from 3.0 to 7.0) of media containing jaggery was varied to study the effect of pH in the fermentation and maximum pullulan yield was obtained at a pH of 5.0. An increase in the initial concentrations (50, 75, 100 g/L) of jaggery in the media produced the maximum pullulan content as 21.6, 19.7 and 18.6 g per 100 g of jaggery, respectively, used. A sucrose based defined media were also used for comparison purposes. Fourier Transform InfraRed (FTIR) spectroscopic analysis was done to confirm the functional groups of synthesized pullulan and compared with that of commercial pullulan.Shake-flask fermentation, under batch cultivation, was investigated for the production of fungal exopolysaccharide, pullulan using jaggery (a traditional concentrated sugar cane juice) as a carbon substrate by Aureobasidium pullulans MTCC 2195. Change in the initial pH (from 3.0 to 7.0) of media containing jaggery was varied to study the effect of pH in the fermentation and maximum pullulan yield was obtained at a pH of 5.0. An increase in the initial concentrations (50, 75, 100 g/L) of jaggery in the media produced the maximum pullulan content as 21.6, 19.7 and 18.6 g per 100 g of jaggery, respectively, used. A sucrose based defined media were also used for comparison purposes. Fourier Transform InfraRed (FTIR) spectroscopic analysis was done to confirm the functional groups of synthesized pullulan and compared with that of commercial pullulan

Anticancer potential of Solanum lycopersicum L. extract in human lung epithelial cancer cells A549

The study aimed to reveal the phytochemical profile, free radical scavenging potential, and anticancer activity of Solanum lycopersicum L. leaf extract (SLLE). According to the study, SLLE contains plant secondary metabolites that are beneficial for health, like phenolics, flavonoids, ascorbic acid, alkaloids, and terpenoids. The SLLE has shown potential free radical scavenging potential in DPPH and ABTS free radical scavenging analysis and its EC50 values (concentration required to inhibit 50% of free radicals) were determined as 481.29 ± 33.82 and 527.56 ± 20.34 µg/mL, respectively. The SLLE has the ability to scavenge free radicals and could be used to treat illnesses brought on by oxidative stress. The anticancer activity of SLLE was assessed by MTT, LDH, micro-morphological, live/dead dual staining, and caspase-3 analysis. In the MTT assay, the IC50 value (concentration required to inhibit 50% of cell viability) of SLLE was determined as 190.41 ± 4.77 µg/mL. Furthermore, SLLE has shown potential anticancer activity by adversely affecting the plasma membrane integrity and escalating the caspase-3 levels. In the biomedical field, SLLE could be highly useful to treat cancer

Application of Syzygium aromaticum, Ocimum sanctum, and Cananga odorata essential oils for management of Ochratoxin A content by Aspergillus ochraceus and Penicillium verrucosum: An in vitro assessment in maize grains

172-182The study is directed to establish the minimizing effects of Syzygium aromaticum, Ocimum sanctum, and Cananga odorata essential oils on the growth and ochratoxin A (OTA) level of Aspergillus ochraceus and Penicillium verrucosum in maize grains. S. aromaticum essential oil (SAEO), O. sanctum essential oil (OSEO), and C. odorata essential oil (COEO) were extracted by hydro-distillation technique, and a total of 50, 44, and 48 chemical constituents were identified by gas chromatography-mass spectrometry (GC-MS), respectively.The SAEO and OSEO belong to the chemotype of eugenol, whereas, COEO was found to be the chemotype of thymol, limonene, and α-ylangene. The antifungal activity of essential oils (EOs) was determined by the micro-well dilution technique. The SAEO showed superior antifungal activity compared to OSEO, COEO, and synthetic antifungal agent nystatin, and its minimum inhibitory concentration (MIC) and minimum fungicidal concentration (MFC) values against A. ochraceous and P. verrucosum were noticed as 1251 ± 42.32 and 1878 ± 28.47 μg/mL, and 0815 ± 22.69 and 1146 ± 51.19 μg/mL, respectively.The antifungal mechanism of EOs was unveiled by assessing the intracellular reactive oxygen species (ROS), ergosterol content, and membrane integrity. The antifungal investigations found that EOs caused fungal mortality by increasing the intracellular ROS, depleting ergosterol synthesis, and distracting membrane integrity. Finally, antifungal and antimycotoxin activity of EOs was demonstrated in maize grains. The SAEO, OSEO, and COEO have reduced the complete fungal growth and OTA level of A. ochraceous and P.verrucosum correspondingly at 2500 and 2500, 3500 and 2500, and 3500 and 3500 μg/g in maize. The EOs could act asnatural antifungal agents; protect foodstuffs from fungal infection and mycotoxins during storage

Development of Monoclonal Antibodies Against Cry1Ac/Ab Protein for Designing of Sandwich ELISA to Detect BT Toxin from Cotton Seeds and Leaves

The design of the study is to develop monoclonal antibodies against Cry1Ac/Ab protein for designing os sandwich ELISA(hybridoma technology). Hybridoma technology was invented by Cesar Milstein and Georges J.F Kohler in the year 1975 and is an unique method used to produce identical antibodies in maximum quantities. Monoclonal antibodies were developed by immunization of Balb/C mice with Cry1Ac/Ab Protein. Titer values of mice tail bleeds were checked and the best mice with higher titer value was used for fusion. Immunized mice spleen cells were fused with Myeloma cells (SP2-O), using polyethylene glycol (PEG) and the fused cells were incubated with HAT medium for 12 days and initially 400 positive hybridoma clones were obtained, of which 13 potential clones were selected using indirect ELISA against Cry1Ac/Ab recombinant antigen. Cross reactivity was ruled out using indriet ELSA against cry proteins such as Cry2A, Cry1F and CP4EPSPS using. Cloning was carried out twice for all 13 clones by limiting dilution factor and pure single clones were selected. The class IgG/IgM/IgA and sub classes IgG1, IgG2, IgG3 antibodies are determined by isotyping. Determination of class and subclass of an antibody is very important for selecting proper purification methods. Commercially available rapid isotyping kits were used for isotyping which provides the information of 1) IgG, IgM, IgA, IgG2a, IgG2b or IgG3 2) Light chain identification as either kappa or lambda. All pure clones were preserved in Liquid Nitrogen for future use to develop immunological kits for detection of Cry1Ac/Ab present in the plant tissue.The design of the study is to develop monoclonal antibodies against Cry1Ac/Ab protein for designing os sandwich ELISA(hybridoma technology). Hybridoma technology was invented by Cesar Milstein and Georges J.F Kohler in the year 1975 and is an unique method used to produce identical antibodies in maximum quantities. Monoclonal antibodies were developed by immunization of Balb/C mice with Cry1Ac/Ab Protein. Titer values of mice tail bleeds were checked and the best mice with higher titer value was used for fusion. Immunized mice spleen cells were fused with Myeloma cells (SP2-O), using polyethylene glycol (PEG) and the fused cells were incubated with HAT medium for 12 days and initially 400 positive hybridoma clones were obtained, of which 13 potential clones were selected using indirect ELISA against Cry1Ac/Ab recombinant antigen. Cross reactivity was ruled out using indriet ELSA against cry proteins such as Cry2A, Cry1F and CP4EPSPS using. Cloning was carried out twice for all 13 clones by limiting dilution factor and pure single clones were selected. The class IgG/IgM/IgA and sub classes IgG1, IgG2, IgG3 antibodies are determined by isotyping. Determination of class and subclass of an antibody is very important for selecting proper purification methods. Commercially available rapid isotyping kits were used for isotyping which provides the information of 1) IgG, IgM, IgA, IgG2a, IgG2b or IgG3 2) Light chain identification as either kappa or lambda. All pure clones were preserved in Liquid Nitrogen for future use to develop immunological kits for detection of Cry1Ac/Ab present in the plant tissue

Biomimetic of hydroxyapatite with Tridax procumbens leaf extract and investigation of antibiofilm potential in Staphylococcus aureus and Escherichia coli

In the last few decades, hydroxyapatite (HA) has become one of the most highly prized biominerals in the biomedical industry for orthopedic and dental applications. The focus of this research was to synthesize biomimetic HA from Tridax procumbens (TP) leaf extract and investigate their antibiofilm properties. The HA was made using the sol-gel method and the HA-TP biocomposite was made by precipitation method. The d.nm size of HA and HA-TP biocomposite was determined as 193.28 and 258.14 d.nm, respectively. The zeta potential of HA and HA-TP biocomposite was determined as −21.2 and −18.3 mV, respectively, and found highly stable. The FTIR study revealed that phytochemicals of TP were successfully impregnated into HA-TP biocomposite. The HA and HA-TP biocomposite were found spherical and agglomerated from SEM analysis. In HR-TEM analysis, the average diameter of the HA and HA-TP biocomposite were 16.57 – 64.22 nm and 51.71 – 138.68 nm, respectively. According to the EDX analysis, HA is primarily composed of calcium, oxygen, and phosphate, whereas, HA-TP biocomposite is primarily composed of calcium, phosphate, oxygen, and carbon. In the antioxidant assay, the IC50 value (concentration required to scavenge 50% of free radicals) of HA-TP biocomposite was determined as 156.69 ± 14.02 and 180.21 ± 12.84 µg/mL in DPPH and ABTS free radical scavenging assays, respectively. The MIC (minimum inhibitory concentration) and MBC (minimum bactericidal concentration) of as-synthesized HA-TP biocomposite against Staphylococcus aureus – ATCC 13565 and Escherichia coli – MTCC 41 were observed as 181.09 ± 21.47 and 317.30 ± 41.03, and 157.59 ± 32.18 and 264.03 ± 21.58 µg/mL, respectively. The as-synthesized HA-TP biocomposite has detrimentally affected the biofilm formation of both the tested bacteria S. aureus – ATCC 13565 and E. coli – MTCC 41. The study concluded that the as-synthesized HA-TP biocomposite could be highly helpful in the biomedical field for alleviating oxidative-stress-related disorders and inhibiting microbial biofilm formation

Hibiscus tiliaceus mediated phytochemical reduction of zinc oxide nanoparticles and demonstration of their antibacterial, anticancer, and dye degradation capabilities

565-574The present research focused on the green, non-toxic, low-cost synthesis of zinc oxide nanoparticles (ZnO NPs) using aqueous extract of Hibiscus tiliaceus leaves as a reducing and stabilizing agent. Thus, synthesized ZnO NPs were characterized by nanotechnological applications, i.e., ultraviolet-visible spectroscopy (UV-vis), dynamic light scattering (DLS), zeta potential, X-ray diffraction (XRD), Fourier transform infrared spectroscopy (FTIR), and high-resolution transmission electron microscopy (HR-TEM). The nanotechnological applications showed that as-synthesized ZnO NPs have bandgap energy of 2.97 eV, zeta potential of 1.2 mV, crystalline in nature (JCPDS data card no-89-1397), and an average size of 30 to 60 nm. The FTIR showed that ZnO NPs have coated with plant secondary metabolites and assisted in the process of green synthesis. The ZnO NPs exhibited broad-spectrum antibacterial activity on Gram-positive and Gramnegative bacteria. The ZnO NPs showed potential anticancer activity against human breast cancer cells MCF-7 and determined the IC50 value as 65.83 ± 2.57 μg/mL by MTT assay. Furthermore, ZnO NPs were used as nano-catalyst for dye degradation of methylene blue, methyl orange, and malachite green with NABH4 as a reducing agent. The ZnO NPs exhibited potent dye degradation capability and followed pseudo-first order kinetics. The study concluded that ZnO NPs could be highly useful as anticancer and antibacterial agents in the biomedical field, and as an environmental cleaning agent for dye degradation in textile industries

Chemical characterization & bioactivity of diketopiperazine derivatives from the mangrove derived Pseudonocardia endophytica

Sediment samples from the mangrove ecosystem of Nizampatnam have been analyzed for actinomycetes as an elite source to screen for the formulation and production of antimicrobial and cytotoxic compounds. The actinomycetes strain VUK-10 has an interesting bioactivity profile and was isolated during our systematic study of mangrove actinomycetes. It was identified as Pseudonocardia endophytica with the aid of polyphasic taxonomy. The ethyl acetate extract of the actinobacterial culture filtrate has been purified by gel-filtration and silica gel column chromatographic purifications led to the isolation of two diketopiperazine compounds, (3S,8aS)-3-isobutylhexahydropyrrolo[1,2-a]pyrazine-1,4-dione (1) and (3R,8aS)-3-isobutylhexahydropyrrolo[1,2-a]pyrazine-1,4-dione (2). The compounds listed, alluring cytotoxic activity against MDA-MB-231, HeLa, MCF-7 and OAW-42 cancer cell lines and also exhibited antimicrobial activities against gram-positive, gram-negative bacteria and fungi. Compound 1 recorded significant antibacterial activity against Xanthomonas campestris and Escherichia coli (8 μg/ml) and compound 2 presented highest activity against Bacillus subtilis (4 μg/ml). Compounds 1 and 2 were active against pathogenic fungi to plants and human beings. The activity was compared with griseofulvin and amphotericin-B, which are standard fungicides. The activity of 16 μg/ml by compound 1 was recorded against Epidermophyton floccosum; an anthropophilic dermatophyte responsible for tinea pedis, tinea cruris and tinea corporis. To the best of our knowledge this is the first narration on the isolation of supra said compounds from the genus Pseudonocardia

Anticancer potential of Solanum lycopersicum L. extract in human lung epithelial cancer cells A549

76-85The study aimed to reveal the phytochemical profile, free radical scavenging potential, and anticancer activity of Solanum lycopersicum L. leaf extract (SLLE). According to the study, SLLE contains plant secondary metabolites that are beneficial for health, like phenolics, flavonoids, ascorbic acid, alkaloids, and terpenoids. The SLLE has shown potential free radical scavenging potential in DPPH and ABTS free radical scavenging analysis and its EC50 values (concentration required to inhibit 50% of free radicals) were determined as 481.29 ± 33.82 and 527.56 ± 20.34 μg/mL, respectively. The SLLE has the ability to scavenge free radicals and could be used to treat illnesses brought on by oxidative stress. The anticancer activity of SLLE was assessed by MTT, LDH, micro-morphological, live/dead dual staining, and caspase-3 analysis. In the MTT assay, the IC50 value (concentration required to inhibit 50% of cell viability) of SLLE was determined as 190.41 ± 4.77 μg/mL. Furthermore, SLLE has shown potential anticancer activity by adversely affecting the plasma membrane integrity and escalating the caspase-3 levels. In the biomedical field, SLLE could be highly useful to treat cancer

Application of Syzygium aromaticum, Ocimum sanctum, and Cananga odorata essential oils for management of Ochratoxin A content by Aspergillus ochraceus and Penicillium verrucosum: An in vitro assessment in maize grains

The study is directed to establish the minimizing effects of Syzygium aromaticum, Ocimum sanctum, and Cananga odorata essential oils on the growth and ochratoxin A (OTA) level of Aspergillus ochraceus and Penicillium verrucosum in maize grains. S. aromaticum essential oil (SAEO), O. sanctum essential oil (OSEO), and C. odorata essential oil (COEO) were extracted by hydro-distillation technique, and a total of 50, 44, and 48 chemical constituents were identified by gas chromatography-mass spectrometry (GC-MS), respectively.The SAEO and OSEO belong to the chemotype of eugenol, whereas, COEO was found to be the chemotype of thymol, limonene, and α-ylangene. The antifungal activity of essential oils (EOs) was determined by the micro-well dilution technique. The SAEO showed superior antifungal activity compared to OSEO, COEO, and synthetic antifungal agent nystatin, and its minimum inhibitory concentration (MIC) and minimum fungicidal concentration (MFC) values against A. ochraceous and P. verrucosum were noticed as 1251 ± 42.32 and 1878 ± 28.47 µg/mL, and 0815 ± 22.69 and 1146 ± 51.19 µg/mL, respectively.The antifungal mechanism of EOs was unveiled by assessing the intracellular reactive oxygen species (ROS), ergosterol content, and membrane integrity. The antifungal investigations found that EOs caused fungal mortality by increasing the intracellular ROS, depleting ergosterol synthesis, and distracting membrane integrity. Finally, antifungal and antimycotoxin activity of EOs was demonstrated in maize grains. The SAEO, OSEO, and COEO have reduced the complete fungal growth and OTA level of A. ochraceous and P. verrucosum correspondingly at 2500 and 2500, 3500 and 2500, and 3500 and 3500 µg/g in maize. The EOs could act as natural antifungal agents; protect foodstuffs from fungal infection and mycotoxins during storage