Green Synthesis of Silver and Titanium Dioxide Nanoparticles Using Euphorbia prostrata Extract Shows Shift from Apoptosis to G0/G1 Arrest followed by Necrotic Cell Death in Leishmania donovani.

The aim of the present study was to synthesize silver (Ag) and titanium dioxide (TiO2) nanoparticles (NPs) using green synthesis from aqueous leaf extract of Euphorbia prostrata as antileishmanial agents and to explore the underlying molecular mechanism of induced cell death. In vitro antileishmanial activity of synthesized NPs was tested against promastigotes of Leishmania donovani by alamarBlue and propidium iodide uptake assays. Antileishmanial activity of synthesized NPs on intracellular amastigotes was assessed by Giemsa staining. The leishmanicidal effect of synthesized Ag NPs was further confirmed by DNA fragmentation assay and by cell cycle progression and transmission electron microscopy (TEM) of the treated parasites. TEM analysis of the synthesized Ag NPs showed a spherical shape with an average size of 12.82 ± 2.50 nm, and in comparison to synthesized TiO2 NPs, synthesized Ag NPs were found to be most active against Leishmania parasites after 24 h exposure, with 50% inhibitory concentrations (IC50) of 14.94 µg/ml and 3.89 µg/ml in promastigotes and intracellular amastigotes, respectively. A significant increase in G0/G1 phase of the cell cycle with a subsequent decrease in S (synthesis) and G2/M phases compared to controls was observed. The growth-inhibitory effect of synthesized Ag NPs was attributed to increased length of S phase. A decreased reactive oxygen species level was also observed, which could be responsible for the caspase-independent shift from apoptosis (G0/G1 arrest) to massive necrosis. High-molecular-weight DNA fragmentation as a positive consequence of necrotic cell death was also visualized. We also report that the unique trypanothione/trypanothione reductase (TR) system of Leishmania cells was significantly inhibited by synthesized Ag NPs. The green-synthesized Ag NPs may provide promising leads for the development of cost-effective and safer alternative treatment against visceral leishmaniasis.

Antiplasmodial activity of eco-friendly synthesized palladium nanoparticles using Eclipta prostrata extract against Plasmodium berghei in Swiss albino mice.

Malaria is an infectious disease caused by the Plasmodium parasite that continues to be a health issue for humans. It is one of the most common pathogenic factors of morbidity and mortality. Palladium nanoparticles (Pd NPs) have been used as target antimicrobial compounds, as a catalyst to manufacture pharmaceuticals, degrade harmful environmental pollutants, and as sensors for the detection of various analyses. The aim of this study was to investigate the antiplasmodial activity of synthesized Pd NPs by using leaf aqueous extract of Eclipta prostrata against Plasmodium berghei in Swiss albino mice. The synthesized Pd NPs were characterized by X-ray diffraction (XRD), Fourier transform infrared (FTIR), Scanning electron microscopy (SEM) with Energy dispersive X-ray spectroscopy (EDX), and High-resolution transmission electron microscope (HRTEM) with the Selected area (electron) diffraction (SAED). The XRD peaks appeared at 35.61°, 44.27°, 56.40°, and 74.51°, which correspond to (111), (200), (220), and (311) planes for palladium, respectively. The FTIR spectra that were carried out to identify the potential biomolecule of synthesized Pd NPs showed the peaks at 3361, 1540, 1399, 1257, 1049, and 659 in the region of 4000-500 cm(-1). The SEM images showed aggregation of NPs with an average size of 63 ± 1.4. The HRTEM images of the precipitated solid phase obtained after termination of the reaction of E. prostrata aqueous leaf extract were in the range from 18 to 64 nm with an average size of 27 ± 1.3 nm. The in vivo antiplasmodial assay was carried out as per Peters' 4-day suppressive test, and the synthesized Pd NP-treated mice group showed reduction of parasitemia by 78.13% with an inhibitory concentration (IC)50 value of 16.44 mg/kg/body weight. The growth inhibition of E. prostrata aqueous leaf extract, palladium acetate, and synthesized Pd NPs showed the IC20, IC50, and IC90 values of 1.90, 10.29, and 64.11; 4.49, 9.84, and 23.04; and 4.34, 8.70, and 18.49 mg/kg/body weight, respectively against NK65 strain of P. berghei. In vitro cytotoxicity of the aqueous leaf extract of E. prostrata, palladium acetate, and Pd NPs that was evaluated against Hep-G2 cell lines showed the cellular toxicity of 7.5, 12, 22, 32, and 39%; 8.2, 18, 32, 55, and 66.2 %; and 8.5, 24, 48, 65, and 76.5% at 1, 10, 100, 250, and 500 µg/mL, respectively. This green chemistry approach toward the synthesis of Pd NPs has many advantages such as, ease with which the process can be scaled up, and economic viability.

Efficacy of larvicidal activity of green synthesized titanium dioxide nanoparticles using Mangifera indica extract against blood-feeding parasites.

Titanium dioxide nanoparticles (TiO2 NPs) are considered to be among the best photocatalytic materials due to their long-term thermodynamic stability, strong oxidizing power, and relative non-toxicity. Nano-preparations with TiO2 NPs are currently under investigation as novel treatments for acne vulgaris, recurrent condyloma acuminata, atopic dermatitis, hyperpigmented skin lesions, and other non-dermatologic diseases. The present study was to investigate the acaricidal and larvicidal activity of synthesized TiO2 NPs utilizing leaf aqueous extract of Mangifera indica L. (Anacardiaceae) against hematophagous parasites. The anti-parasitic activity of TiO2 NPs against the larvae of Rhipicephalus (Boophilus) microplus, Hyalomma anatolicum anatolicum and Haemaphysalis bispinosa (Acari: Ixodidae), fourth instar larvae of Anopheles subpictus, and Culex quinquefasciatus (Diptera: Culicidae) were assessed. The green synthesized TiO2 NPs were analyzed by UV-Vis, FTIR, X-ray diffraction (XRD), AFM, SEM, and TEM. The XRD analysis of synthesized TiO2 NPs revealed the dominant peak at 2θ value of 27.81 which matched the 110 crystallographic plane of the rutile structure indicating the crystal structure. The FTIR spectra exhibited a prominent peak at 3,448 cm(-1) and showed OH stretching due to the alcoholic group, and the OH group may act as a capping agent. The SEM images of TiO2 NPs displayed spherical, oval in shape, individual, and some in aggregates. Characterization of the synthesized TiO2 NPs using AFM offered three-dimensional visualization and uneven surface morphology. The TEM micrograph showed agglomerates, round and slight elongation with an average size of 30 ± 5 nm. The maximum efficacy was observed in synthesized TiO2 NPs against the larvae of R. microplus, Hyalomma anatolicum anatolicum, Haemaphysalis bispinosa, A. subpictus, and Culex quinquefasciatus with LC50 value of 28.56, 33.17, 23.81, 5.84, and 4.34 mg/L, respectively. In the present study, a novel, simple, and eco-friendly approach has been suggested to control blood-feeding parasites.

Solanum trilobatum extract-mediated synthesis of titanium dioxide nanoparticles to control Pediculus humanus capitis, Hyalomma anatolicum anatolicum and Anopheles subpictus.

Titanium dioxide nanoparticles (TiO2 NPs) are widely used in paints, printing ink, rubber, paper, cosmetics, sunscreens, car materials, cleaning air products, industrial photocatalytic processes, and decomposing organic matters in wastewater due to their unique physical, chemical, and biological properties. The present study was conducted to assess the antiparasitic efficacies of synthesized TiO2 NPs utilizing leaf aqueous extract of Solanum trilobatum against the adult head louse, Pediculus humanus capitis De Geer (Phthiraptera: Pediculidae); larvae of cattle tick Hyalomma anatolicum (a.) anatolicum Koch (Acari: Ixodidae), and fourth instar larvae of malaria vector Anopheles subpictus Grassi (Diptera: Culicidae). The green synthesized TiO2 NPs were analyzed by X-ray diffraction (XRD), Fourier transform infrared spectroscopy (FTIR), Scanning electron microscopy (SEM), Energy-dispersive X-ray spectroscopy analysis (EDX), and Atomic force microscopy (AFM). XRD analysis of synthesized TiO2 NPs revealed that the particles were in the form of nanocrystals as evidenced by the major peaks at 2θ values of 27.52°, 36.21°, and 54.43° identified as 110, 101, and 211 reflections, respectively. FTIR spectra exhibited a prominent peak at 3,466 cm(-1) and showed OH stretching due to the alcoholic group, and the OH group may act as a capping agent. SEM images displayed NPs that were spherical, oval in shape, individual, and some in aggregates with an average size of 70 nm. Characterization of the synthesized TiO2 NPs using AFM offered a three-dimensional visualization and uneven surface morphology. The pediculocidal and acaricidal activities of synthesized TiO2 NPs showed the percent mortality of 31, 42, 63, 82, 100; 36, 44, 67, 89, and 100 at 2, 4, 6, 8, and 10 mg/L, respectively, against P. h. capitis and H. a. anatolicum. The average larval percent mortality of synthesized TiO2 NPs was 38, 47, 66, 79, and 100 at 1, 2, 3, 4, and 5 mg/L, respectively, against A. subpictus. The maximum activity was observed in the aqueous leaf extract of S. trilobatum, TiO(OH)2 solutions (bulk), and synthesized TiO2 NPs with LC50 values of 35.14, 25.85, and 4.34 mg/L; 47.15, 29.78, and 4.11 mg/L; and 28.80, 24.01, and 1.94 mg/L, and r (2) values of 0.982, 0.991, and 0.992; 0.947, 0.987, and 0.997; and 0.965, 0.998 and 0.985, respectively, against P. h. capitis, H. a. anatolicum, and A. subpictus. This study provides the first report on the pediculocidal, acaricidal, and larvicidal activity of synthesized TiO2 NPs. This is an ideal eco-friendly, novel, low-cost, and simple approach to satisfy the requirement of large-scale industrial production bearing the advantage for the control of P. h. capitis, H. a. anatolicum, and A. subpictus.

Bioassay-guided isolation and characterization of active antiplasmodial compounds from Murraya koenigii extracts against Plasmodium falciparum and Plasmodium berghei.

Malaria is an overwhelming impact in the poorest countries in the world due to their prevalence, virulence and drug resistance ability. Currently, there is inadequate armoury of drugs for the treatment of malaria. This underscores the continuing need for the discovery and development of new effective and safe antimalarial drugs. To evaluate the in vitro and in vivo antimalarial activity of the leaf ethyl acetate extract of Murraya koenigii, bioassay-guided chromatographic fractionation was employed for the isolation and purification of antimalarial compounds. The in vitro antimalarial activity was assayed by the erythrocytic stages of chloroquine-sensitive strain of Plasmodium falciparum (3D7) in culture using the fluorescence-based SYBR Green I assay. The in vivo assay was done by administering mice infected with Plasmodium berghei (NK65) four consecutive daily doses of the extracts through oral route following Peter's 4-day curative standard test. The percentage suppression of parasitaemia was calculated for each dose level by comparing the parasitaemia in untreated control with those of treated mice. Cytotoxicity was determined against HeLa cells using MTT assay. Histopathology was studied in kidney, liver and spleen of isolated compound-treated Swiss albino mice. The leaf crude ethyl acetate extract of M. koenigii showed good in vitro antiplasmodial activity against P. falciparum. The in vivo test of the leaf crude ethyl acetate extract (600 mg/kg) showed reduced malaria parasitaemia by 86.6% against P. berghei in mice. Bioassay-guided fractionation of the leaf ethyl acetate extract of M. koenigii led to the isolation of two purified fractions C3B2 (2.84 g) and C3B4 (1.97 g). The purified fractions C3B2 and C3B4 were found to be active with IC50 values of 10.5 ± 0.8 and 8.25 ± 0.2 µg/mL against P. falciparum, and in vivo activity significantly reduced parasitaemia by 82.6 and 88.2% at 100 mg/kg/body weight on day 4 against P. berghei, respectively. The isolated fractions C3B2 and C3B4 were monitored by thin-layer chromatography until a single spot was obtained with R f values of 0.36 and 0.52, respectively. The pure compounds obtained in the present investigation were subjected to UV-visible spectroscopy, Fourier transformer infrared spectroscopy, 1D and 2D (1)H-Nuclear magnetic resonance (NMR), (13)C NMR, DEPT, COSY and Mass spectral analysis. Based on the spectral analysis, it is concluded that the isolated compounds were myristic acid (C3B2) and ß-caryophyllene (C3B4). The cytotoxic effect of myristic acid and ß-caryophyllene showed the TC50 values of >100 and 80.5 µg/mL, respectively against HeLa cell line. The histopathology study showed that protection against nephrotoxicity of kidney, hepatic damage of liver and splenocytes protection in spleen was achieved with the highest dose tested at 100 mg/kg/body weight. The present study provides evidence of antiplasmodial compounds from M. koenigii and is reported for the first time.

Marine algae-mediated synthesis of gold nanoparticles using a novel Ecklonia cava.

In the present study, we report rapid biological synthesis of gold nanoparticles (Au NPs) using a novel marine brown alga Ecklonia cava (Family: Lessoniaceae) by the reduction of chloroauric acid. The formation of Au NPs reaction was complete within 1 min at 80 °C and physiochemically characterized with different analytical techniques. FTIR spectroscopy revealed that Au NPs were functionalized with biomolecules that have primary amine group, hydroxyl group and other stabilizing functional groups. X-ray diffraction pattern showed high purity and face-centered cubic structure of Au NPs. Microscopy results showed that these Au NPs are formed with shapes like spherical and triangular with an average size of 30 ± 0.25 nm. Synthesized Au NPs showed good antimicrobial and biocompatibility with human keratinocyte cell line. Thus, physiochemical characteristic results suggest that Au NPs will have promising biomedical applications in different area such as drug delivery, tissue engineering, biosensor, etc.

Antiplasmodial potential of selected medicinal plants from eastern Ghats of South India.

Malaria caused by the protozoan parasite Plasmodium falciparum, is a major health problem of the developing world. In the present study medicinal plants from Eastern Ghats of South India have been extracted with ethyl acetate and assayed for growth inhibition of asexual erythrocytic stages of chloroquine (CQ)-sensitive (3D7) and (CQ)-resistant (INDO) strains of P. falciparum in culture using the fluorescence-based SYBR Green I assay. Studied extracts showed a spectrum of antiplasmodial activities ranging from (a) very good (IC(50)<10-10 µg/mL: Cyperus rotundus and Zingiber officinale); (b) good (IC(50), >10-15 µg/mL: Ficus religiosa and Murraya koenigii); (c) moderate (IC(50)>15-25 µg/mL: Ficus benghalensis); (d) poor activity (IC(50)>25-60 µg/mL) and (e) inactive (IC(50)>60 µg/mL). Resistance indices ranging from 0.78 to 1.28 suggest that some of these extracts had equal promise against the CQ resistant INDO strain of P. falciparum. Cytotoxicity assessment of the extracts against HeLa cell line using MTT assay revealed that the selectivity indices in the range of 3-15 suggesting a good margin of safety.

Entomopathogenic marine actinomycetes as potential and low-cost biocontrol agents against bloodsucking arthropods.

A novel approach to control strategies for integrated blood-feeding parasite management is in high demand, including the use of biological control agents. The present study aims to determine the efficacy of optimized crude extract of actinomycetes strain LK1 as biological control agent against the fourth-instar larvae of Anopheles stephensi and Culex tritaeniorhynchus (Diptera: Culicidae) and adults of Haemaphysalis bispinosa, Rhipicephalus (Boophilus) microplus (Acari: Ixodidae), and Hippobosca maculata (Diptera: Hippoboscidae). Antiparasitic activity was optimized using the Plackett-Burman method, and the design was developed using the software Design-Expert version 8.0.7.1. The production of the optimized crude actinomycetes LK1 strain extract was performed using response surface methodology to optimize the process parameters of protease inhibitor activity of marine actinobacteria for the independent variables like pH, temperature, glucose, casein, and NaCl at two levels (-1 and +1). The potential actinomycetes strain was identified as Saccharomonas spp., and the metamodeling surface simulation procedure was followed. It was studied using a computer-generated experimental design, automatic control of simulation experiments, and sequential optimization of the metamodels fitted to a simulation response surface function. The central composite design (CCD) used for the analysis of treatment showed that a second-order polynomial regression model was in good agreement with the experimental results at R (2) = 0.9829 (p < 0.05). The optimized values of the variables for antioxidant production were pH 6.00, glucose 1.3%, casein 0.09%, temperature 31.23 °C, and NaCl 0.10%. The LK1 strain-optimized crude extract was purified using reversed-phase high-pressure liquid chromatography, and the isolated protease inhibitor showed antiparasitic activity. The antiparasitic activity of optimized crude extract of LK1 was tested against larvae of A. stephensi (LC50 = 31.82 ppm; r(2) = 0.818) and C. tritaeniorhynchus (LC50 = 26.62 ppm; r(2) = 0.790) and adults of H. bispinosa (LC50 = 106.58 ppm; r(2) = 0.871), R. (B.) microplus (LC50 = 92.96 ppm; r(2) = 0.913), and H. maculata (LC50 = 84.90 ppm; r(2) = 0.857).

Eco-friendly microbial route to synthesize cobalt nanoparticles using Bacillus thuringiensis against malaria and dengue vectors.

The developments of resistance and persistence to chemical insecticides and concerns about the non-target effects have prompted the development of eco-friendly mosquito control agents. The aim of this study was to investigate the larvicidal activities of synthesized cobalt nanoparticles (Co NPs) using bio control agent, Bacillus thuringiensis against malaria vector, Anopheles subpictus and dengue vector, Aedes aegypti (Diptera: Culicidae). The synthesized Co NPs were characterized by X-ray diffraction (XRD), Fourier transform infrared (FTIR), Field-emission scanning electron microscopy (FESEM) with energy dispersive X-ray spectroscopy, and Transmission electron microscopy (TEM). XRD analysis showed three distinct diffraction peaks at 27.03°, 31.00°, and 45.58° indexed to the planes 102, 122, and 024, respectively on the face-centered cubic cobalt acetate with an average size of 85.3 nm. FTIR spectra implicated role of the peak at 3,436 cm(-1) for O-H hydroxyl group, 2924 cm(-1) for methylene C-H stretch in the formation of Co NPs. FESEM analysis showed the topological and morphological appearance of NPs which were found to be spherical and oval in shape. TEM analysis showed polydispersed and clustered NPs with an average size of 84.81 nm. The maximum larvicidal mortality was observed in the cobalt acetate solution, B. thuringiensis formulation, and synthesized Co NPs against fourth instar larvae of A. subpictus and A. aegypti with LC50 values of 29.16, 8.12, 3.59 mg/L; 34.61, 6.94, and 2.87 mg/L; r (2) values of 0.986, 0.933, 0.942; 0.962, 0.957, and 0.922, respectively.

Eco-friendly approach using marine actinobacteria and its compounds to control ticks and mosquitoes.

Ticks and mosquitoes are ectoparasitic arthropods that can transmit a variety of diseases to humans and animals during blood feeding and causing serious infectious disorders. The purpose of the present study was to assess the acaricidal and insecticidal property of ethyl acetate extract and its compounds isolated from marine actinobacteria, Streptomyces VITSTK7 sp. against the larvae of cattle ticks, Haemaphysalis bispinosa and Rhipicephalus (Boophilus) microplus (Acari: Ixodidae); fourth-instar larvae of malaria vector, Anopheles subpictus; and filarial vector, Culex quinquefasciatus (Diptera: Culicidae). The ethyl acetate extract was loaded on silica gel column and separated with chloroform, methanol, and acetone as the solvents system. The separation of fractions was visualized by the thin layer chromatography (TLC) plate, further confirmed by high-performance liquid chromatography, and followed by gas liquid chromatography. Three major fractions were analyzed in mass spectroscopy (MS) and matched with existing compounds in the data base. Based on the fragment pattern, it led to the major compounds which were predicted as cyclopentanepropanoic acid, 3,5-bis(acetyloxy)-2-[3-(methoxyimino)octyl], methyl ester (13.3 %) 1; 5-azidomethyl-3-(2-ethoxy carbonyl-ethyl)-4-ethoxycarbonylmethyl-1H-pyrrole-2-carboxylic acid, ethyl ester (18.2 %) 2; and akuammilan-16-carboxylic acid, 17-(acetyloxy)-10-methoxy, methyl ester (16R) (53.3 %) 3. The maximum efficacy was observed in compounds 1, 2, and 3, and the ethyl acetate extract of Streptomyces VITSTK7 sp. against the larvae of H. bispinosa (LC(50) = 1,573.36, 1,333.09, 1,073.29, and 409.71 ppm; r(2) = 0.0.990, 0.934, 0.935, and 0.908), R. microplus (LC(50) = 1,877.86, 815.83, 1,631.14, and 441.54 ppm; r(2) = 0.981, 0.926, 0.0970, and 0.915), A. subpictus (LC(50) = 273.89, 687.69, 464.75, and 223.83 ppm; r(2) = 0.758, 0.924, 0.841, and 0.902), and C. quinquefasciatus (LC(50) = 430.06, 881.59, 777.0, and 195.70 ppm; r(2) = 0.839, 0.859, 0.870, and 0.882), respectively. Results of the present study provide evidence that the maximum parasitic activity of ethyl acetate extract and a synergistic effect of combinations of different compounds have been suggested. The control (distilled water) showed nil mortality in the concurrent assay. In the present study, a novel, targeted, simple, and eco-friendly approach has been suggested to control blood-feeding parasites.

Larvicidal activity of isolated compound 5-(2,4-dimethylbenzyl) pyrrolidin-2-one from marine Streptomyces VITSVK5 sp. against Rhipicephalus (Boophilus) microplus, Anopheles stephensi, and Culex tritaeniorhynchus.

The aim of the present study was to assess the larvicidal property of marine actinobacterial compound 5-(2,4-dimethylbenzyl) pyrrolidin-2-one (DMBPO) extracted and isolated from Streptomyces VITSVK5 sp. tested against the larvae of Rhipicephalus (Boophilus) microplus Canestrini (Acari: Ixodidae), Anopheles stephensi Liston, and Culex tritaeniorhynchus Giles (Diptera: Culicidae). The isolate bacteria was taxonomically characterized, identified, and designated as Streptomyces VITSVK5 sp. The crude extract was loaded on silica gel column and eluted with chloroform:methanol. The isolated pure compound was analyzed by thin layer chromatography using chloroform and methanol as the solvent system and confirmed by high-performance liquid chromatography. The structure of the purified compound was established from infrared, ultraviolet, (1)H-nuclear magnetic resonance (NMR), (13)C-NMR, and mass spectral data. The chemical shift assignments obtained for the aliphatic compound from (1)H-NMR corresponding to the molecular formula C(13)H(17)NO. Bioassay-guided fractionation led to the isolation of compound which was identified as DMBPO. In the present study, Streptomyces VITSVK5 sp. crude extract and different fractions were tested against the larvae of parasites at the concentration of 1,000 ppm. Those fractions showing 100% mortality in 24 h alone was selected for further column chromatographic separation. The purified compound, C(13)H(17)NO, was tested in the concentrations of 500, 250, 125, 62.5, and 31.25 ppm and observed the percent larval mortality of 100, 70, 64, 40, and 28 against R. microplus; 100, 79, 63, 36, and 22 against A. stephensi; and 100, 84, 67, 42, and 27 against C. tritaeniorhynchus, respectively. The crude extract showed parasitic effects after 24 h of exposure at 1,000 ppm, and parasite mortality was observed against the larvae of R. microplus (LC(50) = 210.39 ppm, r (2) = 0.873); A. stephensi (LC(50) = 169.38 ppm, r (2) = 0.840); and C. tritaeniorhynchus (LC(50) = 198.75 ppm, r (2) = 0.887). The maximum efficacy was observed in purified marine actinobacterial compound DMBPO with LC(50) and r (2) values against the larvae of R. microplus (84.31 ppm, 0.889); A. stephensi (88.97 ppm, 0.817), and C. tritaeniorhynchus (74.95 ppm, 0.781), respectively. The control (distilled water) showed nil mortality in the concurrent assay.

Nano-regenerative medicine towards clinical outcome of stem cell and tissue engineering in humans.

Nanotechnology is a fast growing area of research that aims to create nanomaterials or nanostructures development in stem cell and tissue-based therapies. Concepts and discoveries from the fields of bio nano research provide exciting opportunities of using stem cells for regeneration of tissues and organs. The application of nanotechnology to stem-cell biology would be able to address the challenges of disease therapeutics. This review covers the potential of nanotechnology approaches towards regenerative medicine. Furthermore, it focuses on current aspects of stem- and tissue-cell engineering. The magnetic nanoparticles-based applications in stem-cell research open new frontiers in cell and tissue engineering.

Evaluation of antileishmanial activity of South Indian medicinal plants against Leishmania donovani.

Infections due to protozoa of the genus Leishmania are a major worldwide health problem, with high endemicity in developing countries. The aim of this study was to evaluate the in vitro antileishmanial activity of the acetone and methanol leaf extracts of Anisomeles malabarica, flower of Gloriosa superba, leaf of Ocimum basilicum, leaf and seed of Ricinus communis against promastigotes form of Leishmania donovani. Antiparasitic evaluations of different plant crude extracts were performed on 96 well plates at 37°C for 24-48 h. Out of the 10 experimental plant extracts tested, the leaf methanol extracts of A. malabarica, and R. communis showed good antileishmanial activity (IC(50)=126±19.70 and 184±39.33 µg/mL), respectively against promastigotes. Effective antileishmanial activity was observed making these plants as good candidates for isolation of antiprotozoal compounds which could serve as new lead structures for drug development.

Evaluation of stem aqueous extract and synthesized silver nanoparticles using Cissus quadrangularis against Hippobosca maculata and Rhipicephalus (Boophilus) microplus.

The present study was to determine the efficacies of anti-parasitic activities of synthesized silver nanoparticles (Ag NPs) using stem aqueous extract of Cissus quadrangularis against the adult of hematophagous fly, Hippobosca maculata (Diptera: Hippoboscidae), and the larvae of cattle tick, Rhipicephalus (Boophilus) microplus (Acari: Ixodidae). Contact toxicity method was followed to determine the potential of parasitic activity. Twelve milliliters of stem aqueous extract of C. quadrangularis was treated with 88 ml of 1mM silver nitrate (AgNO(3)) solution at room temperature for 30 min and the resulting solution was yellow-brown color indicating the formation extracellular synthesis of Ag NPs. The synthesized Ag NPs were characterized with UV-visible spectroscopy, X-ray diffraction (XRD), Fourier transform infrared spectroscopy (FTIR), Field emission scanning electron microscope (FESEM) and energy dispersive X-ray (EDX) spectroscopy. The synthesized Ag NPs were recorded by UV-visible spectrum at 420 nm and XRD patterns showed the nanoparticles crystalline in nature. FTIR analysis confirmed that the bioreduction of Ag((+)) ions to Ag NPs were due to the reduction by capping material of plant extract. FESEM image of Ag NPs showed spherical and oval in shape. By using the Bragg's Law and Scherrer's constant, the average mean size of synthesized Ag NPs was 42.46 nm. The spot EDX analysis showed the complete chemical composition of the synthesized Ag NPs. The mortality obtained by the synthesized Ag NPs from the C. quadrangularis was more effective than the aqueous extract of C. quadrangularis and AgNO(3) solution (1 mM). The adulticidal activity was observed in the aqueous extract, AgNO(3) solution and synthesized Ag NPs against the adult of H. maculata with LC(50) values of 37.08, 40.35 and 6.30 mg/L; LC(90) values of 175.46, 192.17 and 18.14 mg/L and r(2) values of 0.970, 0.992 and 0.969, respectively. The maximum efficacy showed in the aqueous extract, AgNO(3) solution and synthesized Ag NPs against the larvae of R. (B.) microplus with LC(50) values of 50.00, 21.72 and 7.61 mg/L; LC(90) values of 205.12, 82.99 and 22.68 mg/L and r(2) values of 0.968, 0.945 and 0.994, respectively. The present study is the first report on antiparasitic activity of the experimental plant extract and synthesized Ag NPs. This is an ideal eco-friendly and inexpensive approach for the control of H. maculata and R. (B.) microplus.

Acaricidal efficacy of synthesized silver nanoparticles using aqueous leaf extract of Ocimum canum against Hyalomma anatolicum anatolicum and Hyalomma marginatum isaaci (Acari: Ixodidae).

The use of acaricides had limited efficacy in reducing tick infestations and is often accompanied by serious drawbacks, including the selection of acaricide resistant ticks, contamination of environment, and milk and meat products with drug residues. The present study was based on assessments of the antiparasitic activities to determine the efficacy of synthesized silver nanoparticles (AgNPs) utilizing aqueous leaf extract of Ocimum canum Sims (Labiatae) against the larvae of Hyalomma anatolicum (a.) anatolicum Koch, 1844 and Hyalomma marginatum (m.) isaaci Sharif, 1928 (Acari: Ixodidae). The synthesized AgNPs results were recorded from UV-vis spectrum, X-ray diffraction (XRD) analysis, Fourier transform infrared spectroscopy (FTIR), scanning electron microscopy (SEM) and energy-dispersive X-ray spectroscopy (EDX) analysis. The production of the AgNPs synthesized from the leaf extract of O. canum was evaluated through UV-visible spectrophotometer in a range of wavelength from 300 to 600 nm. This revealed a peak at 426 nm in leaf extracts of O. canum, indicating the production of AgNPs. The XRD spectrum compared with the standard confirmed spectrum of silver particles formed in the present experiments were in the form of nanocrystals, as evidenced by the peaks at 2θ values of 27.71°, 32.16°, 38.08°, 46.15°, 54.70° and 57.35°. The FTIR spectra of AgNPs exhibited prominent peaks at 818, 1,045, 1,381 and 1,616 in the region 500-3,000 cm(-1). The peaks correspond to the presence of a C-H vibration of the aromatic ring, stretch vibration of C-O, carbonyl groups and flavanones. SEM analyses of the synthesized AgNPs were clearly distinguishable, which measured 25-110 nm in size. It is clear that the rod and cylindrical structures have an average size of 95 nm. The EDX spectra showed the purity of the material and the complete chemical composition of the synthesized AgNPs. Parasite larvae were exposed to varying concentrations of aqueous leaf extract of O. canum and synthesized AgNPs for 24 h. The acaricidal activities of aqueous crude leaf extracts of O. canum against the larvae of H. a. anatolicum and H. m. isaaci have LC(50) and LC(90) values of 15.31 and 13.85 mg/L, and 62.41 and 48.86 mg/L, respectively. The efficacies of 1 mM AgNO(3) solution against H. a. anatolicum and H. m. isaaci were LC(50) = 12.25 and 12.17 mg/L, LC(90) = 49.17 and 46.52 mg/L, respectively, and the maximum efficacy was observed in the synthesized AgNPs against H. a. anatolicum and H. m. isaaci with LC(50) and LC(90) values of 0.78 and 1.00 mg/L, and 1.51 and 1.68 mg/L, respectively. This method is considered as an innovative alternative approach to control parasites.

Evaluation of medicinal plant extracts and isolated compound epicatechin from Ricinus communis against Paramphistomum cervi.

The purpose of this study is to determine the efficacies of hexane, chloroform, ethyl acetate, acetone, and methanol leaf extracts of Euphorbia hirta L., Psidium guajava L., Ricinus communis L., Solanum trilobatum L., and Tridax procumbens L. against sheep fluke Paramphistomum cervi (Digenea: Paramphistomatidae). All plant extracts showed moderate effects after 24 h of exposure; however, the highest parasite mortality was found in the methanol extract of R. communis. In the present study, bioassay-guided fractionation of methanol extract of R. communis led to the separation and identification of epicatechin as a potential new compound (LC(50) = 31.2; LC(90) = 105.0 ppm) against P. cervi. The structures were established from infrared, ultraviolet, (1)H-nuclear magnetic resonance (NMR), (13)C-NMR, and mass spectral data which confirmed the identification of the compound epicatechin from R. communis. Results of this study showed that the methanol extract of R. communis may be considered as a potent source and epicatechin as a new natural parasitic agent.

Feeding deterrent activity of synthesized silver nanoparticles using Manilkara zapota leaf extract against the house fly, Musca domestica (Diptera: Muscidae).

With a greater awareness of the hazards associated with the use of synthetic organic insecticides, there has been an urgent need to explore suitable alternative products for pest control. Musca domestica is ubiquitous insect that has the potential to spread a variety of pathogens to humans and livestock. They are mechanical carriers of more than hundred human and animal intestinal diseases and are responsible for protozoan, bacterial, helminthic, and viral infections. The present work aimed to investigate the feeding deterrent activity of synthesized silver nanoparticles (Ag NPs) using leaf aqueous extract of Manilkara zapota against M. domestica. The synthesized Ag NPs were recorded from UV-vis spectrum at 421 nm and scanning electron microscopy confirm the biosynthesis and characterization of Ag NPs with spherical and oval in shape and size of 70-140 nm. The FTIR analysis of the purified nanoparticles showed the presence of bands 1,079, 1,383, 1,627, 2,353, and 2,648 cm(-1), which were complete synthesis of AgNPs; the XRD pattern of AgNPs showed diffraction peaks at 2θ values of 38.06°, 44.37°, 64.51°, and 77.31° sets of lattice planes were observed (111), (200), (220), and (311) facts of silver, respectively. Adult flies were exposed to different concentrations of the aqueous extract of synthesized Ag NPs, 1 mM silver nitrate (AgNO(3)) solution and aqueous extract of M. zapota for 1, 2, and 3 h; however, AgNPs showed 72% mortality in 1 h, 89% mortality was found in 2 h, and 100% mortality was found in 3 h exposure at the concentration of 10 mg/mL and the leaf aqueous extract showed 32% mortality in 1 h, 48% mortality was found in 2 h, and 83% mortality was found in 3 h exposure at concentration of 50 mg/mL. The most efficient activity was observed in synthesized Ag NPs against M. domestica (LD(50) = 3.64 mg/mL; LD(90) = 7.74 mg/mL), the moderate activity reported in the aqueous extract of M. zapota (LD(50) = 28.35 mg/mL; LD(90) = 89.19 mg/mL) and nil activity were observed in AgNO(3) solution at 3 h exposure time at 10 mg/mL. Dimethyl 2, 2-dichlorovinyl phosphate (DDVP) was used as a positive control and showed the LD(50) value of 3.38 mL/L. These results suggest that the synthesized Ag NPs have the potential to be used as an ideal eco-friendly approach for the control of the adult of M. domestica. This method is considered as a new approach to control sanitary pest. Therefore, this study provides first report on the feeding deterrent activity of synthesized Ag NPs against housefly.

Isolation and characterisation of acaricidal and larvicidal novel compound (2S,5R,6R)-2-hydroxy-3,5,6-trimethyloctan-4-one from Streptomyces sp. against blood-sucking parasites.

The aim of the present study was to assess the acaricidal and larvicidal property of marine actinobacterial compound (2S,5R,6R)-2-hydroxy-3,5,6-trimethyloctan-4-one extracted and isolated from Streptomyces sp. VITDDK3 tested against the larvae of Rhipicephalus (Boophilus) microplus Canestrini (Acari: Ixodidae), Anopheles subpictus Grassi and Culex quinquefasciatus Say (Diptera: Culicidae). The isolate was taxonomically characterised, identified and designated as Streptomyces sp. VITDDK3. The crude compound was loaded on silica gel column and eluted with chloroform-methanol-water. The purity of the compound isolated was analysed by thin layer chromatography using chloroform and methanol as the solvent system and confirmed by high-performance liquid chromatography. The structure of the purified compound was established from infrared, ultraviolet, (1)H-nuclear magnetic resonance (NMR), (13)C-NMR and mass spectral data. The chemical shift assignments obtained for the aliphatic compound from (1)H-NMR corresponding to the molecular formula C(11)H(22)O(2). Bioassay-guided fractionation led to the isolation of compound, which was identified as (2S,5R,6R)-2-hydroxy-3,5,6-trimethyloctan-4-one. In the present study, Streptomyces sp. VITDDK3 crude extract and different fractions were tested against the larvae of parasites at a concentration of 1,000 ppm. Those fractions showing 100% mortality in 24 h alone was selected for further column chromatographic separation. The purified compound (2S,5R,6R)-2-hydroxy-3,5,6-trimethyloctan-4-one was tested in the concentrations of 250, 200, 150, 100 and 50 ppm and observed the per cent mortality of 100, 88, 62, 50 and 36 against R. microplus; 100, 100, 87, 62 and 39 against A. subpitcus; and 100, 94, 79, 51 and 33 against C. quinquefasciatus, respectively. The crude extract showed parasitic effects after 24 h of exposure at 1,000 ppm, and parasite mortality was observed against the larvae of R. microplus (LC(50) = 88.74 ppm; r (2) = 0.865) against the larvae of A. subpictus (LC(50) = 162.59 ppm; r (2) = 0.817) and against C. quinquefasciatus (LC(50) = 120.15 ppm; r (2) = 0.782), respectively. The maximum efficacy was observed in purified marine actinobacterial compound (2S,5R,6R)-2-hydroxy-3,5,6-trimethyloctan-4-one with LC(50) and r (2) values against the larvae of R. microplus (94.49 ppm; 0.982) and A. subpictus (69.65 ppm; 0.906) and against C. quinquefasciatus (82.82 ppm; 0.957), respectively. The control (distilled water) showed nil mortality in the concurrent assay.

Antiplasmodial potential of medicinal plant extracts from Malaiyur and Javadhu hills of South India.

The emergence and spread of Plasmodium falciparum with resistance to chloroquine (CQ), the safest and cheapest anti-malarial drug, coupled with the increasing cost of alternative drugs especially in developing countries have necessitated the urgent need to tap the potential of plants for novel anti-malarials. The present study investigates the anti-malarial activity of the methanolic extracts of 13 medicinal plants from the Malaiyur and Javadhu hills of South India against blood stage CQ-sensitive (3D7) and CQ-resistant (INDO) strains of P. falciparum in culture using the fluorescence-based SYBR Green I assay. Sorbitol-synchronized parasites were incubated under normal culture conditions at 2% hematocrit and 1% parasitemia in the absence or presence of increasing concentrations of plant extracts. CQ and artemisinin were used as positive controls, while 0.4% DMSO was used as the negative control. The cytotoxic effects of extracts on host cells were assessed by functional assay using HeLa cells cultured in RPMI containing 10% fetal bovine serum, 0.21% sodium bicarbonate and 50 µg/mL gentamycin (complete medium). Plant extracts (bark methanol extracts of Annona squamosa (IC(50), 30 µg/mL), leaf extracts of Ocimum gratissimum (IC(50), 32 µg/mL), Ocimum tenuiflorum (IC(50), 31 µg/mL), Solanum torvum (IC(50), 31 µg/mL) and Justicia procumbens (IC(50), 63 µg/mL), showed moderate activity. The leaf extracts of Aristolochia indica (IC(50), 10 µg/mL), Cassia auriculata (IC(50), 14 µg/mL), Chrysanthemum indicum (IC(50), 20 µg/mL) and Dolichos biflorus (IC(50), 20 µg/mL) showed promising activity and low activity was observed in the flower methanol extracts of A. indica , leaf methanol extract of Catharanthus roseus, and Gymnema sylvestre (IC(50), >100 µg/mL). These four extracts exhibited promising IC(50) (µg/mL) of 17, 24, 19 and 24 respectively also against the CQ resistant INDO strain of P. falciparum. The high TC(50) in mammalian cell cytotoxicity assay and the low IC(50) in anti-malarial P. falciparum assay indicates selectivity and good resistance indices in the range of 0.9-1.7 for leaf extracts of A. indica, C. auriculata, C. indicum and D. biflorus suggests that these may serve as anti-malarial agents even in their crude form. These results indicate a possible explanation of the traditional use of some of these medicinal plants against malaria or malaria-like conditions.

Efficacy of plant-mediated synthesized silver nanoparticles against hematophagous parasites.

The purpose of the present study was to investigate the acaricidal and larvicidal activity against the larvae of Haemaphysalis bispinosa Neumann (Acarina: Ixodidae) and larvae of hematophagous fly Hippobosca maculata Leach (Diptera: Hippoboscidae) and against the fourth-instar larvae of malaria vector, Anopheles stephensi Liston, Japanese encephalitis vector, Culex tritaeniorhynchus Giles (Diptera: Culicidae) of synthesized silver nanoparticles (AgNPs) utilizing aqueous leaf extract from Musa paradisiaca L. (Musaceae). The color of the extract changed to light brown within an hour, and later it changed to dark brown during the 30-min incubation period. AgNPs results were recorded from UV-vis spectrum at 426 nm; Fourier transform infrared (FTIR) analysis confirmed that the bioreduction of Ag(+) ions to silver nanoparticles are due to the reduction by capping material of plant extract, X-ray diffraction (XRD) patterns clearly illustrates that the nanoparticles formed in the present synthesis are crystalline in nature and scanning electron microscopy (SEM) support the biosynthesis and characterization of AgNPs with rod in shape and size of 60-150 nm. After reaction, the XRD pattern of AgNPs showed diffraction peaks at 2θ = 34.37°, 38.01°, 44.17°, 66.34° and 77.29° assigned to the (100), (111), (102), (110) and (120) planes, respectively, of a faced centre cubic (fcc) lattice of silver were obtained. For electron microscopic studies, a 25 µl sample was sputter-coated on copper stub, and the images of nanoparticles were studied using scanning electron microscopy. The spot EDX analysis showed the complete chemical composition of the synthesized AgNPs. The parasite larvae were exposed to varying concentrations of aqueous extract of M. paradisiaca and synthesized AgNPs for 24 h. In the present study, the percent mortality of aqueous extract of M. paradisiaca were 82, 71, 46, 29, 11 and 78, 66, 38, 31and 16 observed in the concentrations of 50, 40, 30, 20, 10 mg/l for 24 h against the larvae of H. bispinosa and Hip. maculata, respectively. The maximum efficacy was observed in the aqueous extract of M. paradisiaca against the H. bispinosa, Hip. maculata, and the larvae of A. stephensi, C. tritaeniorhynchus with LC(50) values of 28.96, 31.02, 26.32, and 20.10 mg/lm, respectively (r (2) = 0.990, 0.968, 0.974, and 0.979, respectively). The synthesized AgNPs of M. paradisiaca showed the LC(50) and r (2) values against H. bispinosa, (1.87 mg/l; 0.963), Hip. maculata (2.02 mg/l; 0.976), and larvae of A. stephensi (1.39; 0.900 mg/l), against C. tritaeniorhynchus (1.63 mg/l; 0.951), respectively. The χ (2) values were significant at p < 0.05 level.