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.

Bio-functionalized silver nanoparticles: a novel colorimetric probe for cysteine detection.

Chemical interactions between nanoparticles and biomolecules are vital for applying nanoparticles in medicine and life science. Development of sensitive, rapid, low-cost, and eco-friendly sensors for the detection of molecules acting as disease indicator is need of an hour. In the present investigation, a green trend for silver nanoparticle synthesis was followed using leaf extract of Calotropis procera. Silver nanoparticles exhibited surface plasmon absorption peak at 421 nm, spherical shape with average size of 10 nm, and zeta potential of -22.4 mV. The as-synthesized silver nanoparticles were used for selective and sensitive detection of cysteine. Cysteine induces aggregation in stable silver nanoparticles owing to selective and strong interaction of -SH group of cysteine with silver nanoparticle surface. Cysteine-induced silver nanoparticle aggregation can be observed visually by change in color of silver nanoparticles from yellow to pink. Cysteine concentration was estimated colorimetrically by measuring absorption at surface plasmon wavelength. Limit of detection for cysteine using silver nanoparticles is ultralow, i.e., 100 nM. The mechanistic insight into cysteine detection by silver nanoparticles was investigated using FT-IR, TEM, DLS, and TLC analysis. Proposed method can be applied for the detection of cysteine in blood plasma and may give rise to a new insight into development of eco-friendly fabricated nanodiagnostic device in future.

Innovative approach for urease inhibition by Ficus carica extract-fabricated silver nanoparticles: An in vitro study.

In the present study, a rapid, low-cost, and ecofriendly method of stable silver nanoparticles (AgNPs) synthesis using leaves extract of Ficus carica (F. carica), a plant with diverse metabolic consortium, is reported for the first time. An absorption peak at 422 nm in UV-Vis spectroscopy, a spherical shape with an average size of 21 nm in transmission electron microscopy, and crystalline nature in X-ray powder diffraction studies were observed for the synthesized AgNPs. Fourier transform infrared analysis indicated that proteins of F. carica might have a vital role in AgNP synthesis and stabilization. AgNPs were found to inhibit urease, a key enzyme responsible for the survival and pathogenesis of the bacterium, Helicobacter pylori. Inhibition of urease by AgNPs was monitored spectrophotometrically by the evaluation of ammonia release. The urease inhibition potential of AgNPs can be explored in the treatment of H. pylori by preparing novel combinations of standard drugs with AgNPs- or AgNPs-encapsulated drug molecules.

Biofunctionalized silver nanoparticles as a novel colorimetric probe for melamine detection in raw milk.

Nanoparticles have emerged as a promising analytical tool for monitoring food adulteration and safety. In the present study, silver nanoparticles (AgNPs) were synthesized using leaves' extract of Jatropha gossypifolia. AgNPs revealed a characteristic surface plasmon resonance (SPR) peak at 419 nm and have spherical and grain shape with size range between 18 and 30 nm. A selective and rapid method of melamine detection in raw milk was developed with the use of these biofunctionalized AgNPs. The color change, deviation in SPR spectra, and change in the absorption ratio (A500 /A419 ) of AgNPs occurred after an AgNPs-melamine interaction. The detection limit for melamine up to 2 µM (252 ppb) was attained with this method, which is quite lower than safety level recommendations of regulatory bodies demonstrating sensitivity of the method. Dynamicx light scattering and transmission electron microscopy analyses exhibited an increase in hydrodynamic diameter and size of AgNPs after melamine interaction. Melamine sensing by AgNPs was investigated by different physicochemical and thermal analyses.

Mercury sensing and toxicity studies of novel latex fabricated silver nanoparticles.

Safe and eco-friendly alternatives to currently used hazardous chemico-physical methods of silver nanoparticles (AgNPs) synthesis are need of time. Rapid, low cost, selective detection of toxic metals in environmental sample is important to take safety action. Toxicity assessment of engineered AgNPs is essential to avoid its side effects on human and non-target organisms. In the present study, biologically active latex from Euphorbia heterophylla (Poinsettia) was utilized for synthesis of AgNPs. AgNPs was of spherical shape and narrow size range (20-50 nm). Occurrence of elemental silver and crystalline nature of AgNPs was analyzed. Role of latex metabolites in reduction and stabilization of AgNPs was analyzed by FT-IR, protein coagulation test and phytochemical analysis. Latex-synthesized AgNPs showed potential in selective and sensitive detection of toxic mercury ions (Hg(2+)) with limit of detection around 100 ppb. Addition of Hg(2+) showed marked deviation in color and surface plasmon resonance spectra of AgNPs. Toxicity studies on aquatic non-target species Daphnia magna showed that latex-synthesized AgNPs (20.66 ± 1.52% immobilization) were comparatively very less toxic than chemically synthesized AgNPs (51.66 ± 1.52% immobilization). Similarly, comparative toxicity study on human red blood cells showed lower hemolysis (4.46 ± 0.01%) by latex-synthesized AgNPs as compared to chemically synthesized AgNPs causing 6.14 ± 0.01% hemolysis.

Plant extract: a promising biomatrix for ecofriendly, controlled synthesis of silver nanoparticles.

Uses of plants extracts are found to be more advantageous over chemical, physical and microbial (bacterial, fungal, algal) methods for silver nanoparticles (AgNPs) synthesis. In phytonanosynthesis, biochemical diversity of plant extract, non-pathogenicity, low cost and flexibility in reaction parameters are accounted for high rate of AgNPs production with different shape, size and applications. At the same time, care has to be taken to select suitable phytofactory for AgNPs synthesis based on certain parameters such as easy availability, large-scale nanosynthesis potential and non-toxic nature of plant extract. This review focuses on synthesis of AgNPs with particular emphasis on biological synthesis using plant extracts. Some points have been given on selection of plant extract for AgNPs synthesis and case studies on AgNPs synthesis using different plant extracts. Reaction parameters contributing to higher yield of nanoparticles are presented here. Synthesis mechanisms and overview of present and future applications of plant-extract-synthesized AgNPs are also discussed here. Limitations associated with use of AgNPs are summarised in the present review.

Transformation of aromatic dyes using green synthesized silver nanoparticles.

Nowadays, increasing use of nanoproducts in area of human and environmental applications raises concern about safety aspects of nanoparticles synthesized using traditional physicochemical methods. Silver nanoparticles (AgNPs) synthesis at ambient parameters using latex of medicinally important plant Jatropha gossypifolia (J. gossypifolia) is reported in the present study. Potential of AgNPs in degradation of methylene blue and eosin B was also evaluated. Rapid formation of stable AgNPs was analyzed by visual color change from colorless to yellow-red after addition of latex in AgNO3 solution and by characteristic surface plasmon resonance (SPR) peak at 430 nm in UV-Vis spectroscopy. FT-IR analysis, protein coagulation test showed capping of proteins, flavonoids, terpenoids and polyphenols of latex on surface of AgNPs. FE-SEM, HR-TEM analysis revealed spherical shape of AgNPs. Narrow size range of AgNPs (5-40 nm) observed in HR-TEM analysis. EDS analysis confirms the presence of elemental silver while XRD revealed crystalline nature of AgNPs. Zeta potential of -21.4 mV indicates high stability of AgNPs. Effects of different parameters (pH, temperature, incubation time) on nanosynthesis were studied in the present study. Dye reduction studies were performed using UV-Vis spectroscopy, TLC, FT-IR and HPLC analysis showing decreased absorbance maxima of both dyes with respect to time, change in R f values, changes in wave number, transmittance, and retention time of dyes after AgNPs addition. The rate constant for methylene blue and eosin B reduction by AgNPs was found to be 0.062 and 0.022 min(-1).

Catalytic and synergistic antibacterial potential of green synthesized silver nanoparticles: Their ecotoxicological evaluation on Poecillia reticulata.

In the present study, stable silver nanoparticles (AgNPs) were fabricated at a rapid rate from leaf extract of medicinally important plant Alstonia macrophylla. Biosynthesized AgNPs are of spherical shape and narrow size (70 nm), exhibiting a surface plasmon resonance peak at 435 nm, and a zeta potential of -30.8 mV and have a crystalline nature. A diverse biochemical consortium of protein, terpenoids, phenolics, and flavonoids in leaf extract of A. macrophylla was found to be responsible for AgNP synthesis as evidenced from qualitative-quantitative chemical analysis and Fourier transform infrared spectroscopy studies. Nitroaromatic compounds are anthropogenic pollutants with long-lasting environmental persistence and are needed to transform into less toxic derivatives. 4-Nitrophenol and p-nitroaniline were reduced to less hazardous and commercially useful 4-aminophenol and p-phenylenediamine by phytosynthesized AgNPs. Rate constants of 0.052 and 0.040 Min(-1) were calculated for 4-nitrophenol and p-nitroaniline reduction, respectively. Thin-layer chromatography also confirms the reduction of these nitroaromatic compounds. Combinational studies could be one of the strategies to overcome microbial resistance to antibiotics. In synergistic antibacterial assay, the highest increase in a fold area of 3.84 was reported against Staphylococcus aureus using a combination of AgNPs with penicillin. Biosynthesized AgNPs were found to be less toxic (LC50 = 9.13 ppm) than chemically synthesized AgNPs having a LC50 value of 2.86 ppm against nontarget fish Poecillia reticulata. Our green nanosynthesis method offers a faster rate of formation of stable AgNPs having antibacterial and catalytic potential with lower environmental toxicity.

Mosquito Larvicidal Potential of Gossypium hirsutum (Bt cotton) Leaves Extracts against Aedes aegypti and Anopheles stephensi larvae.

BACKGROUND: We aimed to extract the ingredients from leaves of Gossypium hirsutum (Bt cotton) using different solvents and evaluate for potential use to control different larval stages of mosquito species, Aedes aegypti and Anopheles stephensi. METHODS: Qualitative and quantitative estimation of ingredients from Go. hirsutum (Bt) plant extract was carried out and their inhibitory action against mosquito larvae was determined using mosquito larvicidal assay. RESULTS: LC50 values of water, ethanol, ethyl acetate and hexane extracts for Ae. aegypti were 211.73±21.49, 241.64±19.92, 358.07±32.43, 401.03±36.19 and 232.56±26.00, 298.54±21.78, 366.50±30.59, 387.19±31.82 for 4(th) instar of An. stephensi, respectively. The water extract displayed lowest LC50 value followed by ethanol, ethyl acetate and hexane. Owing to the comparatively better activity of water extract, its efficacy was further evaluated for mosquito larvicidal activity, which exhibited LC50 values of 133.95±12.79, 167.65±11.34 against 2(nd) and 3(rd) instars of Ae. aegypti and 145.48±11.76, 188.10±12.92 against 2(nd) and 3(rd) instars of An. stephensi, respectively. Crude protein from the water extract was precipitated using acetone and tested against 2(nd), 3(rd) and 4(th) instars of Ae. aegypti and An. stephensi. It revealed further decrease in LC50 values as 105.72±25.84, 138.23±23.18, 126.19±25.65, 134.04±04 and 137.88±17.59, 154.25±16.98 for 2(nd), 3(rd) and 4(th) instars of Ae. aegypti and An. stephensi, respectively. CONCLUSION: Leaves extracts of Go. hirsutum (Bt) is potential mosquito larvicide and can be used as a potent alternative to chemical insecticides in integrated pest management.

Larvicidal activity of silver nanoparticles synthesized using Pergularia daemia plant latex against Aedes aegypti and Anopheles stephensi and nontarget fish Poecillia reticulata.

In present study, the bioactivity of latex-producing plant Pergularia daemia as well as synthesized silver nanoparticles (AgNPs) against the larval instars of Aedes aegypti and Anopheles stephensi mosquito larvae was determined. The range of concentrations of plant latex (1,000, 500, 250, 125, 62.25, and 31.25 ppm) and AgNPs (10, 5, 2.5, 1.25, 0.625, and 0.3125 ppm) were prepared. The LC(50) and LC(90) values for first, second, third, and fourth instars of synthesized AgNPs-treated first, second, third, and fourth instars of A. aegypti (LC(50) = 4.39, 5.12, 5.66, 6.18; LC(90) = 9.90, 11.13, 12.40, 12.95 ppm) and A. stephensi (LC(50) = 4.41, 5.35, 5.91, 6.47; LC(90) = 10.10, 12.04, 13.05, 14.08 ppm) were found many fold lower than crude latex-treated A. aegypti (LC(50) = 55.13, 58.81, 75.66, 94.31; LC(90) = 113.00, 118.25, 156.95, 175.71 ppm) and A. stephensi (LC(50) = 81.47, 92.09, 96.07, 101.31; LC(90) = 159.51, 175.97, 180.67, 190.42 ppm). The AgNPs did not exhibit any noticeable effects on Poecillia reticulata after either 24 or 48 h of exposure at their LC(50) and LC(90) values against fourth-instar larvae of A. aegypti and A. stephensi. The UV-visible analysis shows absorbance for AgNPs at 520 nm. TEM reveals spherical shape of synthesized AgNPs. Particle size analysis revealed that the size of particles ranges from 44 to 255 nm with average size of 123.50 nm. AgNPs were clearly negatively charged (zeta potential -27.4 mV). This is the first report on mosquito larvicidal activity P. daemia-synthesized AgNPs.

Larvicidal activity of silver nanoparticles synthesized using Plumeria rubra plant latex against Aedes aegypti and Anopheles stephensi.

In the present study activity of silver nanoparticles (AgNPs) synthesized using Plumeria rubra plant latex against second and fourth larval instar of Aedes aegypti and Anopheles stephensi was determined. Range of concentrations of synthesized AgNps (10, 5, 2.5, 1.25, 0.625, 0.3125 ppm) and aqueous crude latex (1,000, 500, 250, 125, 62.50, 31.25 ppm) were tested against larvae of A. aegypti and A. Stephensi. The synthesized AgNps from P. rubra latex were highly toxic than crude latex extract in both mosquito species. The LC(50) values for second and fourth larval instars after 24 h of crude latex exposure were 1.49, 1.82 ppm against A. aegypti and 1.10, 1.74 ppm against A. stephensi respectively. These figures were 181.67, 287.49 ppm against A. aegypti and 143.69, 170.58 ppm against A. stephensi respectively for crude latex extract. The mortality rates were positively correlated with the concentration of AgNPs. The characterization studies of synthesized AgNPs by UV-Vis spectrophotometry, transmission electron microscopy (TEM), Particle size analysis (PSA) and zeta potential confirmed the spherical shape and size (32-200 nm) of silver nanoparticles along with stability. Toxicity studies carried out against non-target fish species Poecilia reticulata, the most common organism in the habitats of A. aegypti and A. stephensi showed no toxicity at LC(50) and LC(90) doses of the AgNPs. This is the first report on mosquito larvicidal activity of latex synthesized nanoparticles.

Insecticidal potency of bacterial species Bacillus thuringiensis SV2 and Serratia nematodiphila SV6 against larvae of mosquito species Aedes aegypti, Anopheles stephensi, and Culex quinquefasciatus.

The tremendous worldwide efforts to isolate novel mosquito larvicidal bacteria with improved efficacy present significant promise to control vector-borne diseases of public health importance. In the present study, two native bacterial isolates, Bacillus thuringiensis (Bt SV2) and Serratia species (SV6) were evaluated for mosquito larvicidal potential against the early fourth instar larvae of Aedes aegypti, Anopheles stephensi, and Culex quinquefasciatus with reference to B. thuringiensis subsp. israelensis (Bti) H 14. The native Gram-positive, spore-forming Bt SV2 isolate showed 100% mortality against early fourth instars of Aedes aegypti, Anopheles stephensi, and Culex quinquefasciatus, in parallel to Bti H14 strain. After 24 h, Bt SV2 showed 98%, 89%, and 80.67%, and Bti H14 showed 92%, 98.33%, and 60% mortality against Aedes aegypti, Anopheles stephensi, and Culex quinquefasciatus, respectively. Serratia SV6 showed highest activity against Culex quinquefasciatus (100%) followed by Anopheles stephensi (95%) and Aedes aegypti (91%) after 48 h of exposure. The Gram-negative Serratia SV6 showed delayed toxicity compared to Bti H14 and Bt SV2 against early fourth instars of Aedes aegypti, Anopheles stephensi, and Culex quinquefasciatus. The relative mortality of all treatments after 12-h exposures showed the varied toxicity with respect to exposure time, bacterial treatment, and mosquito species. Genetic relatedness of the strains was confirmed on the basis of phylogenetic reconstructions based on alignment of 16S rRNA gene sequences which indicated a strong clustering of the strain SV2 with B. thuringiensis and the strain SV6 with Serratia nematodiphila. In conclusion, the native isolate B. thuringiensis SV2 showed significant toxicity while Serratia SV6 showed less and delayed toxicity against several mosquito species compared with BtiH14. They may be used as novel bacterial insecticidal agents in mosquito vector-borne disease control. To our knowledge, this is the first report on mosquito larvicidal potential of Serratia species.

Prodigiosin produced by Serratia marcescens NMCC46 as a mosquito larvicidal agent against Aedes aegypti and Anopheles stephensi.

Microbial control agents offer alternatives to chemical pest control as they can be more selective than chemical insecticides. The present study evaluates the mosquito larvicidal potential of microbial pigment prodigiosin produced by Serratia marcescens NMCC46 against Aedes aegypti and Anopheles stephensi. The pigment of S. marcescens NMCC46 was extracted after 24 h from mannitol containing nutrient broth media. The effects of crude extracted pigment on the growth, survival, development, and other life cycle aspects were studied. The LC(50) and LC(90) values of second, third, and fourth instars of A. aegypti (LC(50) = 41.65, 139.51, 103.95; LC(90) = 117.81, 213.68, 367.82) and A. stephensi (LC(50) = 51.12, 105.52, 133.07; LC(90) = 134.81, 204.45, 285.35) were determined. At higher concentration (500 ppm), mortality starts within first 6 h of exposure. More than 50% mortality occurs within the first 24 h. The overall observed effects against A. aegypti and A. stephensi larvae after 48 h were increasing percent survival larvae, survival pupation, adult emergence with decreasing crude pigment extract concentration. These ensure that the resultant mosquito population reduction is substantial even where the larvicidal potential is minimal. The UV (λ (max) = 536 nm), TLC (Rf = 0.9), HPLC, and FTIR analysis of crude pigment shows the presence of prodigiosin as active compound. Thus, the active compound produced by this species would be more useful against vectors responsible for diseases of public health importance. This is the first report on mosquito larvicidal activity of prodigiosin produced by Serratia species.

Larvicidal potential of silver nanoparticles synthesized using fungus Cochliobolus lunatus against Aedes aegypti (Linnaeus, 1762) and Anopheles stephensi Liston (Diptera; Culicidae).

Larvicides play a vital role in controlling mosquitoes in their breeding sites. The present study was carried out to establish the larvicidal activities of mycosynthesized silver nanoparticles (AgNPs) against vectors: Aedes aegypti and Anopheles stephensi responsible for diseases of public health importance. The AgNPs synthesized by filamentous fungus Cochliobolus lunatus, characterized by UV-Vis spectrophotometry, Fourier transform infrared spectroscopy, X-ray diffraction, scanning electron microscopy, and transmission electron microscopy. The characterization studies confirmed the spherical shape and size (3-21 nm) of silver nanoparticles. The efficacy of mycosynthesized AgNPs at all the tested concentrations (10, 5, 2.5, 1.25, 0.625, and 0.3125 ppm) against second, third, and fourth instar larvae of A. aegypti (LC(50) 1.29, 1.48, and 1.58; LC(90) 3.08, 3.33, and 3.41 ppm) and against A. stephensi (LC(50) 1.17, 1.30, and 1.41; LC(90) 2.99, 3.13, and 3.29 ppm) were observed, respectively. The mortality rates were positively correlated with the concentration of AgNPs. Significant (P < 0.05) changes in the larval mortality was also recorded between the period of exposure against fourth instar larvae of A. aegypti and A. stephensi. The possible larvicidal activity may be due to penetration of nanoparticles through membrane. Toxicity studies carried out against non-target fish species Poecilia reticulata, the most common organism in the habitats of A. aegypti and A. stephensi showed no toxicity at LC50 and LC90 doses of the AgNPs. This is the first report on mosquito larvicidal activity of mycosynthesized nanoparticles. Thus, the use of fungus C. lunatus to synthesize silver nanoparticles is a rapid, eco-friendly, and a single-step approach and the AgNps formed can be potential mosquito larvicidal agents.

Studies on silver accumulation and nanoparticle synthesis By Cochliobolus lunatus.

Development of reliable and eco-friendly processes for synthesis of metallic nanoparticles is an important step in the field of application of nanotechnology. Biological systems provide a useful option to achieve this objective. In this study, potent fungal strain was selectively isolated from soil samples on silver supplemented medium, followed by silver tolerance (100-1,000 ppm) test. The isolated fungus was subjected to morphological, 18S rRNA gene sequencing and phylogenic studies and confirmed as Cochliobolus lunatus. The silver accumulation and nanoparticle formation potential of wet cell mass of C. lunatus was investigated. The accumulation and nanoparticle formation by wet fungal cell mass with respect to pH change was also studied. The desorbing assay was used to recover accumulated silver from cell mass. C. lunatus was found to produce optimum biomass (0.94 g%) at 635 ppm of silver. Atomic absorption spectroscopy study showed that at optimum pH (6.5 ± 0.2), cell mass accumulates 55.6% of 100 ppm silver. SEM and FTIR studies revealed that the cell wall of C. lunatus is the site of silver sorption, and certain organic groups such as carbonyl, carboxyl, and secondary amines in the fungal cell wall have an important role in biosorption of silver in nanoform. XRD determined the FCC crystalline nature of silver nanoparticles. TEM analysis established the shape of the silver nanoparticles to be spherical with the presence of very small-sized nanoparticles. Average size of silver nanoparticles (14 nm) was confirmed by particle sizing system. This study reports the synthesis and accumulation of silver nanoparticles through reduction of Ag(+) ions by the wet cell mass of fungus C. lunatus.

Studies on characterization of bioflocculant exopolysaccharide of Azotobacter indicus and its potential for wastewater treatment.

Partially characterized bioflocculant exopolysaccharide (EPS) produced from an Azotobacter indicus ATCC 9540 strain reported in our previous study was further characterized, and its flocculant potential was investigated at different pH, temperature, and cations concentrations. Flocculant activity at different concentrations of EPS in the absence of cations was reanalyzed by slight modified flocculant assay. It revealed that flocculant activity increased in a concentration-dependent manner up to a certain limit, with the maximum flocculation of 72% at 500 mg L⁻¹ EPS concentration, even in the absence of cations. At the concentration of 10 mg L⁻¹, CaCl2 showed more significant activity (92%) than AlCl3 and MnSO4. Differential scanning calorimetry study and flocculant assay revealed high temperature stability of EPS up to 97 °C. Molecular weight of the EPS determined by size exclusion chromatography was found to be approximately 2 x 106 kDa. Investigation on flocculation efficacy of the characterized EPS for wastewater treatment of dairy, woolen, starch, and sugar industry suggested it to be effective and stable at wide pH range of 5-10. Wastewater treatment with biopolymer at 500 mg L⁻¹ showed reduction in biochemical oxygen demand (38-80%), chemical oxygen demand (37-79%), and suspended solids (41-68%). This study suggests that Azotobacter polymer has high potential in wastewater treatment as bioflocculant and can be used as a potential alternative to chemical flocculants.

Bioefficacy of Plumbago zeylanica (Plumbaginaceae) and Cestrum nocturnum (Solanaceae) plant extracts against Aedes aegypti (Diptera: Culicide) and nontarget fish Poecilia reticulata.

In a search for natural products that could be used to control the vectors of tropical diseases, extracts of medicinal plants Plumbago zeylanica and Cestrum nocturnum have been tested for larvicidal activity against second, third, and fourth instar larvae of Aedes aegypti. The LC(50) values of all the extracts in different solvents of both the plants were less than 50 ppm (15.40-38.50 ppm) against all tested larval instars. Plant extracts also affected the life cycle of A. aegypti by inhibition of pupal development and adult emergence with increasing concentrations. The larvicidal stability of the extracts at five constant temperatures (19°C, 22°C, 25°C, 28°C, and 31°C) evaluated against fourth instar larvae revealed that toxicity of both plant extracts increases with increase in temperature. Toxicity studies carried out against fish species Poecilia reticulata, the most common nontarget organism in the habitats of A. aegypti, showed almost nil to meager toxicity at LC(50) and LC(90) doses of the plant extracts. The qualitative analysis of crude extracts of P. Zeylanica and C. nocturnum revealed the presence of bioactive phytochemicals with predominance of plumbagin in P. zeylanica and saponins in C. nocturnum. Partially purified plumbagin from P. zeylanica and saponins from C. nocturnum were obtained, and their presence was confirmed by thin-layer chromatography and biochemical tests. The bioassay experiment of partially purified secondary metabolites showed potent mosquito larvicidal activity against the fourth instar larval form. Therefore, this study explored the safer and effective potential of plant extracts against vector responsible for diseases of public health importance.

Bioflocculant exopolysaccharide production by Azotobacter indicus using flower extract of Madhuca latifolia L.

Efficacy of Azotobacter indicus ATCC 9540 strain for production exopolysaccharide (EPS) bioflocculant was investigated. Mahua flower extract (Madhuca latifolia L), a natural substrate at the concentration of 20 g L(-1), gave maximum recovery of EPS followed by sucrose and mannitol as compared to other carbon sources after 172 h. Yeast extract was found to be the most effective nitrogen source as compared to beef extract, sodium nitrate, ammonium sulfate, casein hydrolysate, and urea for the production of EPS. EPS production was increased in presence of nitrogen (5.51 g L(-1)) as compared to nitrogen-free medium (3.51 g L(-1)), and fermentation time was also reduced by 28 h. Maximum EPS production (6.10 g L(-1)) was found in the presence of 20 g L(-1) flower extract and 0.5 g L(-1) yeast extract containing Ashby's media with 180 rpm at 30 degrees C at 144 h, under controlled conditions in 2.5 L fermenter using optimized medium. The isolated EPS showed cation-dependent flocculating activity. Concentration of EPS played an important role in bioflocculating activity which increased in a concentration-dependent manner up to a certain limit, with the maximum flocculation of 72% at 500 mg L(-1) concentration but remained almost static after this concentration. Extracted polymer was characterized by different chemical tests, FT-IR spectroscopy, and TLC which showed presence of uronic acids, O-acetyl groups, and Orcinol with suggestive indication of alginate like polymer. This study suggests that use of M. latifolia L. flowers can be a potential alternative bioresource for production of exopolysaccharide.