Improved naringinase double screen plate assay: progress towards the perfect screening.

Naringinase is an important enzyme for commercial purposes due to its dual activity as both α-l-rhamnosidase and ß-d-glucosidase. The traditional method for screening microbes that produce naringinase involves growing them on naringin agar, but this method has limitations and result in false positive results. This is because the growth on the naringin agar plate could be due to the presence of other organisms that produce rhamnosidase or other glucosidases, or those that use agar as a carbon source, rather than actual naringinase producers. To address these limitations, a double screen plate assay was developed using synthetic substrates, to separately test for ß-d-glucosidase and α-l-rhamnosidase activity. The presence of a yellow zone of p-nitrophenol indicates the action of these enzymes, and the intensity of the yellow colour zone indicates the potential for naringinase production. This new screening method is a significant improvement in identifying real naringinase producers and represents progress towards a more reliable screening assay.

Naringenin biosynthesis and fabrication of naringenin mediated nano silver conjugate for antimicrobial potential.

The development of resistance, instability and high doses are some drawbacks of biologically active natural products. Modification of natural compounds to make it broad spectrum is the standard approach in drug design. This paper sets to modify the naringenin by silver nanoparticle conjugation to enhance its already reported pharmacological activities. The naringenin-nano silver conjugate was synthesized by one-step green synthesis, that is, sunlight exposure confirmed by UV spectroscopy. The biosynthesized naringenin-nanosilver conjugate was tested for antiacanthamoebal and antimicrobial potential. The antibacterial potential was increased by 5.8-6.14 fold against Gram positive bacteria, that is, S. aureus and Bacillus subtilis and 4.5-13.6 fold against Gram negative bacteria, that is, Escherichia coli and Pseudomonas aeruginosa. The standard naringenin-nanosilver conjugate significantly reduced the LC50 values against the Acanthamoeba cells, by, 66% and 36%, as compared to substrate naringin and standard naringenin respectively while biotransformed naringinin-nanosilver conjugate reduced LC50 by 50.56%, compared with biotransformed naringenin. Hence modification of natural product as nanoconjugate is the best practice for improvement as an effective drug.

Rhizobium pusense-Mediated Selenium Nanoparticles-Antibiotics Combinations against Acanthamoeba sp.

Severe ocular infections by Acanthamoeba sp. lead to keratitis, resulting in irreversible vision loss in immune-compromised individuals. When a protozoal infection spreads to neural tissues, it causes granulomatous encephalitis, which can be fatal. Treatment often takes longer due to the transition of amoeba from trophozoites to cyst stages, cyst being the dormant form of Acanthamoeba. A prolonged use of therapeutic agents, such as ciprofloxacin (Cipro), results in severe side effects; thus, it is critical to improve the therapeutic efficacy of these widely used antibiotics, possibly by limiting the drug-sensitive protozoal-phase transition to cyst formation. Owing to the biomedical potential of selenium nanoparticles (SeNPs), we evaluated the synergistic effects of ciprofloxacin and Rhizobium pusense-biogenic SeNPs combination. SeNPs synthesized using Rhizobium pusense isolated from root nodules were characterized using UV-Visible spectrophotometer, FT-IR, SEM with EDX, particle size analysis, and Zeta potential. The combination was observed to reduce the sub-lethal dose of Cipro, which may help reduce its side effects. The selenium and ciprofloxacin (SeNPs-Cipro) combination reduced the LC50 by 33.43%. The anti-protozoal efficacy of SeNPs-Cipro was found to transduce through decreased protozoal-cyst formations and the inhibition of the galactosidase and protease enzymes of trophozoites. Furthermore, high leakage of sugar, proteins, and amino acids during the SeNPs-Cipro treatment was one primary reason for killing the trophozoites. These experimental results may be helpful in the further pre-clinical evaluation of SeNPs-Cipro to combat protozoal infections. Future studies for combinations of SeNPs with other antibiotics need to be conducted to know the potential of SeNPs against antibiotic resistance in Acanthamoeba.

Moringa Tree, Gift of Nature: a Review on Nutritional and Industrial Potential.

Purpose of Review: Worldwide occurring Moringa plant is commonly famous as a fruit vegetable, known as drumstick or shevga all over India. The miraculous nutritional potential of the drumstick plant was already proved by worldwide research. But in the common population, it is unknown for the nutritional potential of its leaves. The majority of the population is known it only as a fruit vegetable. The Moringa leaves contain almost all essential nutrients, growth factors, vitamins, amino acids, proteins, minerals, and metals like potassium, iron, and zinc. Besides these, nowadays, plant leaves may be used to prepare various nutritional supplements and medicine. Recent Findings: Besides this, this review takes into account some joint efforts of NASI, Allahabad-funded project to use these Moringa leaves for different formulations and its popularization efforts for malnutrition eradication in tribal, i.e., development of recipes of Moringa leaves that will not only make easy preparations but also help to make habitual use of Moringa leaves today. Summary: This review describes the morphology, occurrence, and distribution of Moringa sp., chemical constitutions of Moringa leaves, its potential as anticancer, antidiabetes, and antimicrobial agent and as a nutritional supplement and the commercial future of various products.

Enzymatic response of Moina macrocopa to different sized zinc oxide particles: An aquatic metal toxicology study.

Zinc oxide particles (ZnOPs) of both nanometer and sub-micron sizes are important components of high demand consumer products such as sunscreen, paint, textile, food packaging, and agriculture. Their ultimate discharge in the aquatic ecosystem is nearly unavoidable. For sustainable use of ZnOPs, there is an urgent need to assess its ecotoxicity using ecological indicator organisms. Moina macrocopa, an important component of the aquatic ecosystem is one such less explored indicator organism. In the present investigation, ZnOPs of two different sizes (250 ± 20 and 500 ± 50 nm) were selected for risk assessment as most of the previous reports were based on the use of 10-100 nm ZnOPs. ZnOPs of 500 nm were more lethal than that of 250 nm size, with respective LC50 of 0.0092 ± 0.0012 and 0.0337 ± 0.0133 mg/L against M. macrocopa after 48 h of exposure. We further used a sublethal concentration of 500 nm (0.00336 mg/L) and 250 nm (0.00092 mg/L) ZnOPs followed by measurement of enzymatic biomarkers of toxicity (acetylcholinesterase, digestive enzymes, antioxidant enzymes). A size-dependent variation in enzymatic response to 250 and 500 nm ZnOPs was seen. Exposure to ZnOPs inhibited acetylcholinesterase and digestive enzymes (trypsin, amylase), and elevated antioxidant enzymes (catalase, glutathione S-transferase) levels. The exposure also decreased the superoxide dismutase activity and increased that of ß-galactosidase. Microscopic investigation revealed the accumulation of ZnOPs in the digestive tract of M. macrocopa that possibly disrupts enzyme activities. The present study will contribute to establishing regulatory policy on the maximum permissible limit of ZnOPs in different water bodies.

Bacterial Pigment Prodigiosin Demonstrates a Unique Antiherpesvirus Activity That Is Mediated through Inhibition of Prosurvival Signal Transducers.

Herpes simplex virus (HSV) is among the most prevalent viral infections worldwide and remains incurable. While nucleoside analogs are used to relieve symptoms of infection, they suffer from having serious adverse effects and are unable to abolish the virus from the host. Here, we demonstrate a unique antiviral effect of prodigiosin (PG), a natural secondary metabolite produced by Serratia marcescens, on HSV infection. We show that PG naturally exerts antiviral activity against HSV-1 and HSV-2 infections. PG treatment resulted in robust inhibition of viral replication in vitro and ex vivo in cultured porcine corneas. Additionally, PG protected against HSV-1 infection and disease progression in a murine model of ocular infection. In our quest to determine the molecular mechanisms of its antiviral activity, we show that PG specifically inhibits NF-κB and Akt signaling pathways and promotes accelerated cell death in HSV-infected cells. Our findings reveal novel antiviral properties of PG, suggesting its high potential as an alternative treatment for herpetic diseases. They also provide new information on antiviral effects of HSV-bacterial metabolite interactions.IMPORTANCE In this article, we provide a new role for a commonly found bacterial pigment in controlling herpes simplex virus infection, for which diverse and multimodal antiviral agents are needed to prevent drug resistance. Serratia marcescens is a red pigment (prodigiosin)-producing Gram-negative bacillus that is naturally found in soil and water. It is associated with many kinds of human infections, including wound and eye infections, and meningitis. Taking cues from previous studies on prodigiosin, including possible proapoptotic anticancer properties, we investigated how it might affect HSV infection. Interestingly, we found that it is a potent virucidal compound that disrupts host signaling pathways needed for HSV growth and survival. The mode of antiviral action suggests potentially broad activity against enveloped viruses. Our results also indicate that interactions with commensal bacteria may inhibit HSV infection, underscoring the importance of studying these microbial metabolites and their implications for viral pathogenesis and treatment.

Production and characterization of multifacet exopolysaccharide from an agricultural isolate, Bacillus subtilis.

This study aims to explore the fermentative production and physicochemical properties of an exopolysaccharide (EPS) produced from agricultural isolate, Bacillus subtilis S1 in submerged culture. The structural characterization (Ultraviolet-visible spectroscopy, Fourier transform infrared spectroscopy, and 13 C Nuclear magnetic resonance spectrometry) revealed that the EPS is an acidic heteropolymer consisting of glucose, glucuronic acid, pyruvic acid, and succinic acid. The non-Newtonian shear thickening nature of EPS with a 1.55 × 107  Da molecular weight is confirmed by rheology analysis. The extracted EPS was 61.3% amorphous with partial crystallinity (38.7%) as confirmed by X-ray diffraction analysis. The EPS shows two-step decomposition and thermal stability up to 300 °C as confirmed by thermogravimetric analysis and differential scanning calorimetry analysis. The EPS has a small Z-average particle size (74.29 nm), high porosity (92.99%), high water holding (92.39%), and absorption capacity (1,198%). The biocompatible nature is confirmed by cytotoxic testing on the human keratinocytes cell line. The demonstrated unique characteristics of Bacillus EPS presents it as a choice of biomaterial for diverse applications.

Nano-eco toxicity study of gold nanoparticles on aquatic organism Moina macrocopa: As new versatile ecotoxicity testing model.

In the field of nanoecotoxicology, very few reports have focused on biochemical changes in non-target organisms after nanoexposure. A less explored aquatic non-target crustacean, Moina macrocopa, was used in the present study to analyze toxicity effects of gold nanoparticles (AuNPs), an emerging nanomaterial. AuNPs was fabricated using tannic acid and were 29 ± 2 nm in size. The 48 h LC50 value of AuNPs was 14 ± 0.14 mg/L against M. macrocopa. The sub-lethal exposure of M. macrocopa juveniles to AuNPs (1.47 and 2.95 mg/L) decreased the activities of acetyl cholinesterase and digestive enzymes (trypsin and amylase). A concentration dependant increase in the activities of antioxidant enzymes such as catalase, superoxide dismutase and glutathione S-transferase suggested the generation of oxidative stress in M. macrocopa after AuNPs exposure. Changes in enzyme activity can be utilized as biomarker(s) for early detection of nanoparticle contamination in aquatic habitat. AuNPs accumulation in gut of M. macrocopa increased the metal bio burden (11 mg/L) and exhibited inhibitory action on digestive enzymes. Complete depuration of AuNPs was not observed after transferring nano-exposed M. macrocopa to normal medium without AuNPs. AuNPs tended to adhere on external body parts such as setae, carapace of M. macrocopa which interfered with swimming activity and also changed the behavioral pattern. AuNPs underwent agglomeration in the medium used for maintenance of M. macrocopa. As nanomaterials are emerging pollutants in aquatic systems, the present work highlights the hazardous effect of AuNPs and development of enzymatic biomarkers to curtail it at community level.

A novel screening method for potential naringinase-producing microorganisms.

Naringinase has high industrial importance, and the progress in naringinase research is still quite slow. The unavailability of an effective, simple screening method, which will be applicable to different microorganisms such as bacteria, fungi, and actinomycetes, is one of the main reasons for this gap. Therefore, a simple plate assay was developed for effective screening of microorganisms for naringinase by exposing to iodine vapors. This plate assay will fill the technological void for simple screening method and will lead to screen more potent industrially important naringinase-producing microorganisms.

Moina macrocopa as a non-target aquatic organism for assessment of ecotoxicity of silver nanoparticles: Effect of size.

The release of nanomaterials in water reservoirs is hazardous. Very few reports are available on the interaction of different sized nanoparticles with aquatic organisms and aquatic environment. In the present study, silver nanoparticles (AgNPs) having an average particle size of 20.80 ±â€¯2.31 and 40.04 ±â€¯4.72 nm were synthesized using polyvinylpyrrolidone and l-tyrosine. Ecotoxicological effects of AgNPs were evaluated on less explored crustacean species, Moina macrocopa. The 48 h lethal values (48 h LC50) of 20 and 40 nm AgNPs were 0.11 ±â€¯0.02 and 0.12 ±â€¯0.03 mg/L, respectively. Further, a size dependent inhibition of AgNPs on acetyl cholinesterase and digestive enzymes (trypsin, amylase, ß-galactosidase) was observed, while that of the antioxidant enzymes (catalase, superoxide dismutase, glutathione-S-transferase) and alkaline phosphatase were enhanced as compared to control group. These results strengthen the potential of enzymes as biomarker in environmental risk assessment of AgNPs. AgNPs accumulated in the gut of M. macrocopa which could not be completely eliminated, thereby resulting in an increased metal body burden. The accumulation of AgNPs of 20 nm was lower than that of 40 nm indicating the influence of size of nanoparticles on uptake and toxicity. AgNPs agglomerated in moderately hard water medium (MHWM) and this agglomeration influenced the exposure the organism thereto. The size of AgNPs influenced the toxicity to M. macrocopa through interplay between uptake, accumulation, aggregation, and excretion in the organism and environment.

Heavy Metal Stress and Its Consequences on Exopolysaccharide (EPS)-Producing Pantoea agglomerans.

Currently, the heavy metal pollution is of grave concern, and the part of microorganism for metal bioremediation should take into account as an efficient and economic strategy. On this framework, the heavy metal stress consequences on exopolysaccharide (EPS)-producing agricultural isolate, Pantoea agglomerans, were studied. The EPS production is a protective response to stress to survive and grow in the metal-contaminated environment. P. agglomerans show tolerance and mucoid growth in the presence of heavy metals, i.e., mercury, copper, silver, arsenic, lead, chromium, and cadmium. EDX first confirmed the metal accumulation and further, FTIR determined the functional groups involved in metal binding. The ICP-AES identified the location of cell-bound and intracellular metal accumulation. Metal deposition on cell surface has released more Ca2+. The effect on bacterial morphology investigated with SEM and TEM revealed the sites of metal accumulation, as well as possible structural changes. Each heavy metal caused distinct change and accumulated on cell-bound EPS with some intracellular deposits. The metal stress caused a decrease in total protein content and increased in total carbohydrate with a boost in EPS. Thus, the performance of P. agglomerans under metal stress indicated a potential candidate for metal bioremediation. Graphical Abstract ᅟ.

Extracellular red Monascus pigment-mediated rapid one-step synthesis of silver nanoparticles and its application in biomedical and environment.

The development of a safe and eco-friendly method for metal nanoparticle synthesis has an increasing demand, due to emerging environmental and biological harms of hazardous chemicals used in existing nanosynthesis methods. The present investigation reports a rapid one-step, eco-friendly and green approach for the formation of nanosized silver particles (AgNPs) using extracellular non-toxic-colored fungal metabolites (Monascus pigments-MPs). The formation of nanosized silver particles utilizing Monascus pigments was confirmed after exposure of reaction mixture to sunlight, by visually color change and further established by spectrophotometric analysis. The size, shape, and topography of synthesized MPs-AgNPs were well-defined using different microscopic and spectroscopic techniques, i.e., FE-SEM, HR-TEM, and DLS. The average size of MPs-AgNPs was found to be 10-40 nm with a spherical shape which was highly stable and dispersed in the solution. HR-TEM and XRD confirmed crystalline nature of MPs-AgNPs. The biocidal potential of MPs-AgNPs was evaluated against three bacterial pathogens such as Pseudomonas aeruginosa, Escherichia coli, and Staphylococcus aureus and it was observed that the MPs-AgNPs significantly inhibited the growth of all three bacterial pathogens. The anti-biofilm activity of MPs-AgNPs was recorded against antibiotic-resistant P. aeruginosa. Besides, the colorimetric metal sensing using MPs-AgNPs was studied. Among the metals tested, the selective Hg2+-sensing potential at micromolar concentration was observed. In conclusion, this is the rapid one-step (within 12-15 min), environment-friendly method for synthesis of AgNPs and synthesized MPs-AgNPs could be used as a potential antibacterial agent against antibiotic-resistant bacterial pathogens.

Prospective of Microbial Exopolysaccharide for Heavy Metal Exclusion.

Metals as a resource are depleting, and on another side, it fetches serious environmental pollution causing a threat to human health and ecosystem. The heavy metal accumulation due to anthropogenic activities results in toxicological manifestation. The traditional methods of remediation are not cost effective, efficient, and ecofriendly which necessitate and motivate towards the safe, effective, and ecofriendly biological methods. The increasing presence of heavy metals in the microbial habitat compels the microbes to develop the ability to tolerate or resist the presence of heavy metals. Exopolysaccharide (EPS) production is one of the strategies of microbes to fight against metal stress. EPS is a microbial biopolymer which is generally produced under stress from harsh environment and nutrition conditions. EPSs are cell-associated or secreted outside the cell and comprised organic macromolecules such as polysaccharides, proteins, and phospholipids in addition to some non-polymeric molecules. EPSs work as competent biosorbents with an anionic reactant group that effectively sequesters cationic heavy metals by electrostatic interactions. The present paper summarizes the EPSs with its types, role, and biosynthesis and an endeavor to elucidate the interaction mechanism of EPSs with heavy metal with supportive and distinctive applications for heavy metal exclusion. The review concluded with the current challenges and future prospects to make the EPS an efficient biosorbent.

Mechanistic approach for fabrication of gold nanoparticles by Nitzschia diatom and their antibacterial activity.

The problem of chemically synthesized nanoproducts motivated scientific community to explore ecofriendly methods of nanosynthesis. Diatoms belong to a group of aquatic, unicellular, photosynthetic microalgae have been scarcely investigated as a source of reducing and capping agents for nanosynthesis of pesticides and antibiotics. The present study reports a novel ecofriendly method for the fabrication of bioactive gold nanoparticles using locally isolated Nitzschia diatoms. The diatom-fabricated gold nanoparticles show characteristic ruby red colored with sharp absorbance peak at 529 nm. Electron microscopy confirmed irregular shape of gold nanoparticles, with average size of 43 nm and zeta potential of -16.8 mV. The effects of gold nanoparticles on diatom viability were investigated using light and electron microscopy. The mechanistic approach to shed light on how diatoms reacted after exposure to gold metal salt revealed that exposure to gold chloride triggers elevated levels of catalase and peroxidase (12.76 and 14.43 unit/mg protein, respectively) to relieve reactive oxygen species (ROS) stress induced by gold salt exposure. Investigation studies on mechanisms behind Nitzschia-mediated gold nanoparticles fabrication outlined the role of diatom proteins, polysaccharides in reduction, and stabilization of nanoparticles as confirmed by FT-IR analysis. Bioactivity of gold nanoparticles was accessed by coupling them with antibiotics (penicillin and streptomycin), which increased their antibacterial activity compared to individual nanoparticles and antibiotics (Escherichia coli, Pseudomonas aeruginosa, and Staphylococcus aureus). Overall, the present novel phyco-nanotechnological approach is a promising tool to be used as sustainable strategy in green nanotechnology as well as to reduce use of antibiotics in microbial control.

Fluconazole treatment enhances extracellular release of red pigments in the fungus Monascus purpureus.

Traditional methods for the production of food grade pigments from the fungus Monascus spp. mostly rely on submerged fermentation. However, the cell-bound nature and intracellular accumulation of pigments in Monascus spp. is a major hurdle in pigment production by submerged fermentation. The present study focused on the investigation of the effect of the antifungal agent fluconazole on red pigment production from Monascus purpureus (NMCC-PF01). At the optimized concentration of fluconazole (30 µg ml-1), pigment production was found to be enhanced by 88% after 96 h and it remained constant even after further incubation up to 168 h. Ergosterol, a sterol specific to fungi, was also extracted and estimated as a function of fungal growth. The concentration of ergosterol in fluconazole-treated fermentation broth was reduced by 49% as compared to control broth. Thus it could be responsible for facilitating the release of intracellular and cell-bound pigments. Nevertheless, the role of cell transporters in transporting out the red pigments cannot be ignored and deserves further attention. Qualitative analysis of red pigment by thin layer chromatography, UV spectroscopy and mass spectrometric analysis (ESIMS) has confirmed the presence of the well-known pigment rubropunctamine. In addition, this fermentation process produces citrinin-free pigments. This novel approach will be useful to facilitate increased pigment production by the release of intracellular or cell-bound Monascus pigments.

Antimicrobial activity of prodigiosin is attributable to plasma-membrane damage.

The bacterial pigment prodigiosin has various biological activities; it is, for instance, an effective antimicrobial. Here, we investigate the primary site targeted by prodigiosin, using the cells of microbial pathogens of humans as model systems: Candida albicans, Escherichia coli, Staphylococcus aureus. Inhibitory concentrations of prodigiosin; leakage of intracellular K+ ions, amino acids, proteins and sugars; impacts on activities of proteases, catalases and oxidases; and changes in surface appearance of pathogen cells were determined. Prodigiosin was highly inhibitory (30% growth rate reduction of C. albicans, E. coli, S. aureus at 0.3, 100 and 0.18 µg ml-1, respectively); caused leakage of intracellular substances (most severe in S. aureus); was highly inhibitory to each enzyme; and caused changes to S. aureus indicative of cell-surface damage. Collectively, these findings suggest that prodigiosin, log Poctanol-water 5.16, is not a toxin but is a hydrophobic stressor able to disrupt the plasma membrane via a chaotropicity-mediated mode-of-action.

Trypsin inactivation by latex fabricated gold nanoparticles: A new strategy towards insect control.

Before applying nanotechnologies in biomedical and environmental areas it is advised to study interactions of nanoparticles and other nanomaterials with biomacromolecule present in living system. Moreover there is scarcity of reports on interactions between nanoparticles and biomaterials. In present report a rapid, ecofriendly method of fabricating stable gold nanoparticles (AuNPs) using latex of Jatropha curcas is reported for the first time. AuNPs found to have characteristic absorption maxima centered at 540nm, multiple irregular shapes with size range from 20 to 50nm and have crystalline nature. Latex fabricated AuNPs were found to inhibit catalytic potential of trypsin (a vital enzyme responsible for digestion, insecticide resistance and in several disease conditions). The interactions between AuNPs and trypsin were analyzed by UV-vis spectrophotometry and microwave plasma-atomic emission spectrometry which suggests formation of trypsin-AuNPs complex responsible for lowering catalytic activity of trypsin. Transmission electron microscopy, Fourier transform infrared spectroscopy and particle size distribution studies further confirm complex formation between trypsin and AuNPs. Diverse interactions of metal nanoparticles with proteins such as covalent interaction, electrostatic interactions and binding to SH group of amino acid may be the reasons behind inhibition of trypsin activity. In vivo studies on serum of several vectors and agriculturally important pests supported instrumental results on AuNPs induced trypsin inhibition. This work will bring a new research direction to explore eco-friendly nanoparticle in insect control via inhibition of enzyme catalytic potential.

In situ development of nanosilver-impregnated bacterial cellulose for sustainable released antimicrobial wound dressing.

PURPOSE: Bacterial cellulose (BC) is an interesting biomaterial found application in various fields due to its novel characteristics like purity, water holding capacity, degree of polymerization and mechanical strength. BC as wound dressing material has limitation because it has no antimicrobial activity. To circumvent this problem, the present study was carried out by impregnation of silver on bacterial cellulose surface. METHODS: Bacterial cellulose was produced by Gluconoacetobacter hansenii (strain NCIM 2529) by shaking culture method. The sodium borohydride and classical Tollens reaction was used for silver nanoparticle synthesis. RESULTS: The effectiveness of sodium borohydride method compared with Tollens reaction was evaluated on the basis of silver nanoparticle formation and its impregnation on BC as evidenced by UV-Vis spectrum analysis, FE-SEM-EDS analysis and FT-IR spectrum. The potential of nano silver impregnated BC was determined for sustained release antimicrobial wound dressing material by swelling ratio, mechanical properties and antimicrobial activity against Staphylococcus aureus. CONCLUSIONS: Thus the nanosilver impregnated bacterial cellulose as promising antimicrobial wound dressing material was evidenced.

Mosquito larvicidal and pupaecidal potential of prodigiosin from Serratia marcescens and understanding its mechanism of action.

Mosquitoes spread lethal diseases like malaria and dengue fever to humans. Considering mosquito vector control as one of the best alternatives to reduce new infections, here we have analyzed the effect of purified pigment prodigiosin extracted from Serratia marcescens (NMCC 75) against larval and pupal stages of Aedes aegypti and Anopheles stephensi mosquitoes. Mosquito larvicidal activities of purified prodigiosin revealed LC50 values of 14 ± 1.2, 15.6 ± 1.48, 18 ± 1.3, 21 ± 0.87 µg/ml against early IInd, IIIrd, IVth instar and pupal stages of Ae. aegypti, respectively. LC50 values for An. stephensi were found to be 19.7 ± 1.12, 24.7 ± 1.47, 26.6 ± 1.67, 32.2 ± 1.79 µg/ml against early IInd, IIIrd, IVth instar and pupae of An. stephensi, respectively. Further investigations toward understanding modes of action revealed variations in the activities of esterases, acetylcholine esterases, phosphatases, proteases and total proteins in the fourth instar larvae of Ae. aegypti indicating intrinsic difference in biochemical features due to prodigiosin treatment. Although there was no inhibition of enzymes like catalase and oxidase but may have profound inhibitory effect on carbonic anhydrase or H(+)-V-ATPase which is indicated by change in the pH of midgut and caeca of mosquito larvae. This reduced pH may be possibly due to the proton pump inhibitory activity of prodigiosin. Pure prodigiosin can prove to be an important molecule for mosquito control at larval and pupal stages of Ae. aegypti and An. stephensi. This is the first report on the mosquito pupaecidal activity of prodigiosin and its possible mechanism of action.

Effect of different carbon sources on morphology and silver accumulation in Cochliobolus lunatus.

The morphology of filamentous fungi plays very important role in uptake of metabolites and enzyme production. A filamentous fungus may be fibrous, hyphae, pellets, clumps, etc. Cochliobolus lunatus is a fungus which has previously been reported for silver accumulation and nanoparticles formation. The present study investigated the role of various carbon sources on morphology, biochemical profile, silver accumulation, and biosynthesis of silver nanoparticles by fungal strain C. lunatus. In this investigation, effect of different carbon sources was studied on morphology of C. lunatus and its silver accumulating ability. As a result of different carbon sources like carboxymethyl cellulose (CMC), pectin, starch, agar, sucrose, and mannitol, the organism showed three kinds of morphologies like homogenous smooth branched clumps, tough short fibrous filaments, and tough pellets, as well as silver accumulating ability. Atomic absorption spectroscopy (AAS) studies showed maximum uptake of Ag(+): 87.44 ± 0.23 and 82.57 ± 0.19 % in pectin- and CMC-grown biomass, respectively. The crystalline nature of silver nanoparticles (AgNPs) was confirmed by X-ray diffraction studies. Transmission electron microscopy (TEM) micrographs of silver nanoparticles confirmed size ranging from 5 to 38 nm.