Phytoconstituents Assisted Biofabrication of Copper Oxide Nanoparticles and Their Antiplasmodial, and Antilarval Efficacy: A Novel Approach for the Control of Parasites.

The present work aimed to biofabricate copper oxide nanoparticles (CuO NPs) using Tinospora cordifolia leaf extract. The biofabricated CuO NPs were treated against the malarial parasite of chloroquine-resistant Plasmodium falciparum (INDO) and the antilarval efficacy was evaluated against the malaria vector Anopheles stephensi and dengue vector Aedes aegypti. The prominence at 285 nm in the UV-visible spectrum helped to identify the produced CuO NPs. Based on the XRD patterns, the concentric rings correspond to reflections at 38.26° (111), 44.11° (200), 64.58° (220), and 77.34° (311). These separations are indicative of CuO's face-centered cubic (fcc) structure. The synthesized CuO NPs have FTIR spectra with band intensities of 3427, 2925, 1629, 1387, 1096, and 600 cm-1. The absorbance band at 3427 cm-1 is known to be associated with the stretching O-H due to the alcoholic group. FTIR proved that the presence of the -OH group is responsible for reducing and capping agents in the synthesis of nanoparticles (NPs). The synthesized CuO NPs were found to be polymorphic (oval, elongated, and roughly spherical) in form with a size range of 11-47 nm and an average size of 16 nm when the morphology was examined using FESEM and HRTEM. The highest antiplasmodial efficacy against the chloroquine-resistant strain of P. falciparum (INDO) was found in the synthesized CuO NPs, with LC50 values of 19.82 µg/mL, whilst HEK293 cells are the least toxic, with a CC50 value of 265.85 µg/mL, leading to a selectivity index of 13.41. However, the antiplasmodial activity of T. cordifolia leaf extract (TCLE) and copper sulfate (CS) solution showed moderate activity, with LC50 values of 52.24 and 63.88 µg/mL, respectively. The green synthesized NPs demonstrated extremely high antilarval efficacy against the larvae of An. stephensi and Ae. aegypti, with LC50 values of 4.06 and 3.69 mg/L, respectively.

Toxicity studies of nanofabricated palladium against filariasis and malaria vectors.

The present study was carried out to establish the biofabrication of palladium nanoparticles (PdNPs) using the plant leaf extract of Tinospora cordifolia Miers and its toxicity studies on the larvae of filariasis vector, Culex quinquefasciatus Say and malaria vector, Anopheles subpictus Grassi. The biofabricated PdNPs were characterized by using UV-visible spectrum, FTIR, XRD, FESEM, EDX and HRTEM. HRTEM confirmed the PdNPs were slightly agglomerated and spherical in shape and the average size was 16 nm. Gas chromatography and mass spectrometry analysis result revealed that the major constituent present in the T. cordifolia leaf extract is 2,4-di-tert-butylphenol (31.79%) whereas the minor compounds are 1-hexadecanol (7.97%), 1-octadecanol (7.70%), 1-eicosanol (6.85%), behenic alcohol (5.36%), 1-tetradecene (6.22%), cyclotetradecane (6.23%), 1-hexadecene (7.97%), 1-octadecene (7.70%), 1-eicosene (6.85%), and 1-docosene (5.36%). T. cordifolia leaf extract exhibited the larvicidal activity against the fourth instar larvae of C. quinquefasciatus and A. subpictus with the values of LC50 = 59.857 and 54.536 mg/L; LC90 = 113.445 and 108.940 mg/L, respectively. The highest toxicity was observed in the biofabricated PdNPs against the fourth instar larvae of C. quinquefasciatus and A. subpictus with the values of LC50 = 6.090 and 6.454 mg/L; LC90 = 13.689 and 13.849 mg/L, respectively. Concerning non-target effects, Poecilia reticulata were exposed to PdNPs for 24 h and did not exhibit any noticeable toxicity. Overall, our findings strongly suggest that PdNPs is a perfect ecological and inexpensive approach for the control of filariasis and malaria vectors.

Evaluation of antiplasmodial activity of medicinal plants from North Indian Buchpora and South Indian Eastern Ghats.

BACKGROUND: Development of resistance against the frontline anti-malarial drugs has created an alarming situation, which requires intensive drug discovery to develop new, more effective, affordable and accessible anti-malarial agents. METHODS: Inspired by their ethnobotanical reputation for being effective against febrile diseases, antiplasmodial potential of ethyl acetate extracts (EAE) and methanol extracts (ME) of 17 medicinal plants collected from the Eastern Ghats of South India and Buchpora, North India were explored against Plasmodium falciparum in vitro using the SYBR Green assay. The results were validated both by confirmation that the fall in fluorescence signal was not due to quenching effects mediated by phytochemical extracts and by Giemsa-stained microscopy. RESULTS: Using EAE or ME, promising antiplasmodial activity (IC50 Pf3D7 ≤ 20 µg/ml), was seen in Aerva lanata (Whole aerial parts-EAE), Anisomeles malabarica (Leaf-EAE), Anogeissus latifolia (bark-EAE), Cassia alata (leaves-EAE), Glycyrrhiza glabra (root-EAE), Juglans regia (seed-ME), Psidium guajava (leaf-ME and EAE) and Solanum xanthocarpum (Whole aerial parts-EAE). EAEs from leaves of Couroupita guianensis, Euphorbia hirta, Pergularia daemia, Tinospora cordifolia and Tridax procumbens as also ME from Ricinus communis (leaf and seed) showed good antiplasmodial activity (Pf 3D7 IC50 21 - 40 µg/ml). Moderate activity (Pf 3D7 IC50: 40-60 µg/mL) was shown by the leaf EAEs of Cardiospermum halicacabum, Indigofera tinctoria and Ricinus communis while the remaining extracts showed marginal (Pf 3D7 IC50 60 to >100 µg/ml) activities. The promising extracts showed good resistance indices (0.41 - 1.4) against the chloroquine resistant INDO strain of P. falciparum and good selectivity indices (3 to > 22.2) when tested against the HeLa cell line. CONCLUSION: These results provide validity to the traditional medicinal usage of some of these plants and further make a case for activity-guided purification of new pharmacophores against malaria.

Green synthesis of titanium dioxide nanoparticles using Psidium guajava extract and its antibacterial and antioxidant properties.

OBJECTIVE: To determine the efficacies of antibacterial and antioxidant activities of aqueous leaf extract of Psidium guajava mediated biosynthesis of titanium dioxide nanoparticles (TiO2 NPs). METHODS: Synthesized TiO2 NPs were tested by disc diffusion method against against human pathogenic bacteria. The total antioxidant activity and phenolic content (Folin-Ciocalteau method) of synthesized TiO2 NPs and aqueous plant extract were determined. The scavenging radicals were estimated by DPPH method. The synthesized TiO2 NPs were characterized by XRD, FTIR, FESEM and EDX. RESULTS: FTIR spectra of synthesized TiO2 NPs exhibited prominent peaks at 3 410 cm(-1) (alkynes), 1 578 cm(-1), 1 451 cm(-1) (alkanes), and 1 123 cm(-1)(C-O absorption). The morphological characterization of synthesized TiO2 NPs was analysed by FESEM which showed spherical shape and clusters with an average size of 32.58 nm. The maximum zone of inhibition was observed in the synthesized TiO2 NPs (20 µg/mL) against Staphylococcus aureus (25 mm) and Escherichia coli (23 mm). The synthesized TiO2 NPs showed more antibacterial activity than the standard antibiotic disk, tetracycline which drastically reduces the chances for the development of antibiotics resistance of bacterial species. The plant aqueous extract and synthesized TiO2 NPs were found to possess maximum antioxidant activity when compared with ascorbic acid. The content of phenolic compounds (mg/g) in leaf aqueous extract and synthesized TiO2 NPs were found to be 85.4 and 18.3 mgTA/g, respectively. CONCLUSIONS: Green synthesized TiO2 NPs provides a promising approach can satisfy the requirement of large-scale industrial production bearing the advantage of low-cost, eco-friendly and reproducible.

Effect of sub-acute exposure to nickel nanoparticles on oxidative stress and histopathological changes in Mozambique tilapia, Oreochromis mossambicus.

The aim of the present study was to assess the oxidative stress, antioxidant response and histopathological changes of nickel nanoparticles (Ni NPs) exposure (14 days) in Mozambique tilapia, Oreochromis mossambicus. Ni NPs were synthesized by metal salt reduction method and characterized by X-ray diffraction (XRD) and Transmission electron microscopy (TEM). The XRD peaks at 44°, 51° and 76° were indexed to the (111), (200) and (220) Bragg's reflections of cubic structure of Nickel, respectively. The crystallite sizes were calculated using Scherrer's formula applied to the major intense peaks and found to be the size of 56nm. TEM images showed that the synthesized Ni NPs are spherical in shape. Biochemical analysis indicated that the superoxide dismutase (SOD), catalase (CAT) and peroxidase (POD) activity was significantly affected by Ni NPs treated O. mossambicus. Reduced antioxidant enzymes and the contents of antioxidants were lowered in the liver and gills of fishes treated with Ni NPs. After 14 days of exposure, a significant accumulation of Ni in the Ni NPs in experimental group was observed in the gill and skin tissues, with the highest levels found in the liver. Ni NPs exposed fish showed nuclear hypertrophy (NH), nuclear degeneration (ND), necrosis (NC) and irregular-shaped nuclei were observed in liver tissue. The hyperplasia of the gill epithelium (GE), lamellar fusion of secondary lamellae (LF), dilated marginal channel (MC), epithelial lifting (EL) and epithelial rupture were observed in gill tissue. Degeneration in muscle bundles (DM), focal area of necrosis (NC) vacuolar degeneration in muscle bundles (VD), edema between muscle bundles (ED) and splitting of muscle fibers were noticed in skin tissue. Further ecotoxicological evaluation will be made concerning the risk of Ni NPs on aquatic environment.

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.

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.

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.

Acaricidal activity of synthesized titanium dioxide nanoparticles using Calotropis gigantea against Rhipicephalus microplus and Haemaphysalis bispinosa.

OBJECTIVE: To assess the acaricidal activity of titanium dioxide nanoparticles (TiO2 NPs) synthesized from flower aqueous extract of Calotropis gigantea(C. gigantea) against the larvae of Rhipicephalus (Boophilus) microplus [R. (B.) microplus] and the adult of Haemaphysalis bispinosa (H. bispinosa). METHODS: The lyophilized C. gigantea flower aqueous extract of 50 mg was added with 100 mL of TiO(OH)2 (10 mM) and magnetically stirred for 6 h. Synthesized TiO2 NPs were characterized by X-ray diffraction (XRD), Fourier transform infrared spectroscopy (FTIR), Scanning electron microscopy (SEM), and Energy dispersive X-ray spectroscopy (EDX). The synthesised TiO2 NPs were tested against the larvae of R. (B.) microplus and adult of H. bispinosa were exposed to filter paper impregnated method. RESULTS: XRD confirmed the crystalline nature of the nanoparticles with the mean size of 10.52 nm. The functional groups for synthesized TiO2 NPs were 1 405.19, and 1 053.45 cm(-1) for -NH2 bending, primary amines and amides and 1 053.84 and 1 078.45 cm(-1) for C-O. SEM micrographs of the synthesized TiO2 NPs showed the aggregated and spherical in shape. The maximum efficacy was observed in the aqueous flower extract of C. gigantea and synthesized TiO2 NPs against R. (B.) microplus (LC50=24.63 and 5.43 mg/L and r(2)=0.960 and 0.988) and against H. bispinosa (LC50= 35.22 and 9.15 mg/L and r(2) = 0.969 and 0.969), respectively. CONCLUSIONS: The synthesized TiO2 NPs were highly stable and had significant acaricidal activity against the larvae of R. (B.) microplus and adult of H. bispinosa. This study provides the first report of synthesized TiO2 NPs and possessed excellent anti-parasitic activity.

Biological approach to synthesize TiO2 nanoparticles using Aeromonas hydrophila and its antibacterial activity.

Nanosized materials have been an important subject in basic and applied sciences. A novel, low-cost, green and reproducible bacteria, Aeromonas hydrophila mediated biosynthesis of titanium dioxide nanoparticles (TiO2 NPs) was reported. The resulting nanoparticles were characterized by FTIR, XRD, AFM and FESEM with EDX. FTIR showed characteristic bands (1643 and 3430 cm(-1)) finds the role of carboxyl group OH stretching amine NH stretch in the formation of TiO2 NPs. The XRD spectrum confirmed that the synthesized TiO2 NPs were in the form of nanocrystals, as evidenced by the peaks at 2θ values of 27.47°, 31.77°, 36.11°, 41.25°, 54.39°, 56.64° and 69.54° were identified as 110, 100, 101, 111, 211, 220 and 301 reflections, respectively. The crystallite sizes were calculated using Scherrer's formula applied to the major intense peaks and found to be the size of 40.50 nm. The morphological characterization was analyzed by FESEM and the analysis showed the NPs smooth shaped, spherical and uneven. GC-MS analysis showed the main compounds found in A. hydrophila were uric acid (2.95%), glycyl-L-glutamic acid (6.90%), glycyl-L-proline (74.41%) and L-Leucyl-D-leucine (15.74%). The potential glycyl-L-proline could have played an important role as a capping agent. A possible mechanism for the biosynthesis of TiO2 NPs has been proposed. The antibacterial activity of the synthesized TiO2 NPs was assessed by well diffusion method toward A. hydrophila, Escherichia coli, Pseudomonas aeruginosa, Staphylococcus aureus, Streptococcus pyogenes and Enterococcus faecalis and showed effective inhibitory activity against S. aureus (33 mm) and S. pyogenes (31 mm).

Novel and simple approach using synthesized nickel nanoparticles to control blood-sucking parasites.

The present study was on assessment of the anti-parasitic activities of nickel nanoparticles (Ni NPs) against the larvae of cattle ticks Rhipicephalus (Boophilus) microplus and Hyalomma anatolicum (a.) anatolicum (Acari: Ixodidae), fourth instar larvae of Anopheles subpictus, Culex quinquefasciatus and Culex gelidus (Diptera: Culicidae). The metallic Ni NPs were synthesized by polyol process from Ni-hydrazine as precursor and Tween 80 as both the medium and the stabilizing reagent. The synthesized Ni NPs were characterized by Fourier transform infrared (FTIR) spectroscopy analysis which indicated the presence of Ni NPs. Synthesized Ni NPs showed the X-ray diffraction (XRD) peaks at 42.76°, 53.40°, and 76.44°, identified as 111, 220, and 200 reflections, respectively. Scanning electron microscopy (SEM) analysis of the synthesized Ni NPs clearly showed that the Ni NPs were spherical in shape with an average size of 150 nm. The Ni NPs showed maximum activity against the larvae of R. (B.) microplus, H. a. anatolicum, A. subpictus, C. quinquefasciatus and C. gelidus with LC(50) values of 10.17, 10.81, 4.93, 5.56 and 4.94 mg/L; r(2) values of 0.990, 0.993, 0.992, 0.950 and 0.988 and the efficacy of Ni-hydrazine complexes showed the LC(50) values of 20.35, 22.72, 8.29, 9.69 and 7.83 mg/L; r(2) values of 0.988, 0.986, 0.989, 0.944 and 0.978, respectively. The findings revealed that synthesized Ni NPs possess excellent larvicidal parasitic activity. To the best of our knowledge, this is the first report on larvicidal activity of blood feeding parasites using synthesized Ni NPs.

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.

Antimalarial activities of medicinal plants traditionally used in the villages of Dharmapuri regions of South India.

ETHNOPHARMACOLOGICAL RELEVANCE: An ethnopharmacological investigation of medicinal plants traditionally used to treat diseases associated with fevers in Dharmapuri region of South India was undertaken. Twenty four plants were identified and evaluated for their in vitro activity against Plasmodium falciparum and assessed for cytotoxicity against HeLa cell line. AIM OF THE STUDY: This antimalarial in vitro study was planned to correlate and validate the traditional usage of medicinal plants against malaria. MATERIALS AND METHODS: An ethnobotanical survey was made in Dharmapuri region, Tamil Nadu, India to identify plants used in traditional medicine against fevers. Selected plants were extracted with ethyl acetate and methanol and evaluated for antimalarial activity against erythrocytic stages of chloroquine (CQ)-sensitive 3D7 and CQ-resistant INDO strains of Plasmodium falciparum in culture using the fluorescence-based SYBR Green I assay. Cytotoxicity was determined against HeLa cells using MTT assay. RESULTS: Promising antiplasmodial activity was found in Aegle marmelos [leaf methanol extract (ME) (IC(50)=7 µg/mL] and good activities were found in Lantana camara [leaf ethyl acetate extract (EAE) IC(50)=19 µg/mL], Leucas aspera (flower EAE IC(50)=12.5 µg/mL), Momordica charantia (leaf EAE IC(50)=17.5 µg/mL), Phyllanthus amarus (leaf ME IC(50)=15 µg/mL) and Piper nigrum (seed EAE IC(50)=12.5 µg/mL). The leaf ME of Aegle marmelos which showed the highest activity against Plasmodium falciparum elicited low cytotoxicity (therapeutic index>13). CONCLUSION: These results provide validation for the traditional usage of some medicinal plants against malaria in Dharmapuri region, Tamil Nadu, India.

Acaricidal, insecticidal, and larvicidal efficacy of fruit peel aqueous extract of Annona squamosa and its compounds against blood-feeding parasites.

Plant products may be alternative sources of parasitic control agents, since they constitute a rich source of bioactive compounds that are eco-friendly and nontoxic products. The plant extracts are good and safe alternatives due to their low toxicity to mammals and easy biodegradability. In the present study, fruit peel aqueous extract of Annona squamosa (Annonaceae) extracted by immersion method exhibited adulticidal activity against Haemaphysalis bispinosa (Acarina: Ixodidae) and the hematophagous fly, Hippobosca maculata (Diptera: Hippoboscidae), and larvicidal activity against the cattle tick Rhipicephalus (Boophilus) microplus (Acari: Ixodidae), Anopheles subpictus, and Culex quinquefasciatus (Diptera: Culicidae). The chemical composition of A. squamosa fruit peel aqueous extract was analyzed by gas chromatography-mass spectrometry. The major chemical constituent of peel aqueous extract of A. squamosa was identified as 1H- cycloprop[e]azulen-7-ol decahydro-1,1,7-trimethyl-4-methylene-[1ar-(1aα,4aα, 7ß, 7 a, ß, 7bα)] (28.55%) by comparison of mass spectral data and retention times. The other major constituents present in the aqueous extract were retinal 9-cis- (12.61%), 3,17-dioxo-4-androsten-11alpha-yl hydrogen succinate (6.86%), 1-naphthalenepentanol decahydro-5-(hydroxymethyl)-5,8a-dimethyl-y,2-bis(methylene)-(1α,4aß,5α,8aα) (14.83%), 1-naphthalenemethanol decahydro -5-(5-hydroxy-3-methyl-3-pentenyl)- 1,4a-di methyl - 6-methylene -(1S-[1α, 4aα, 5α(E), 8aß] (4.44%), (-)-spathulenol (20.75%), podocarp-7-en-3-one13ß-methyl-13-vinyl- (5.98%), and 1-phenanthrene carboxaldehyde 7-ethenyl-1,2,3,4,4a,4,5,6,7,9,10,10a-dodecahydro-1,4a,7-trimethyl-[1R-(1α,4aß.4bα,7ß, 10aα)]-(5.98%). The adult and larval parasitic mortalities observed in fruit peel aqueous extract of A. squamosa were 31, 59, 80, 91, and100%; 27, 42, 66, 87, and 100%; and 33, 45, 68, 92, and 100% at the concentrations of 250, 500, 1,000, 1,500, and 2,000 ppm, respectively, against Haemaphysalis bispinosa, Hippobosca maculata, and R. microplus. The observed larvicidal efficacies were 36, 55, 72, 92, 100% and 14, 34, 68, 89, and 100% at 200, 400, 600, 800, and 1,000 ppm, respectively, against A. subpictus and C. quinquefasciatus. The highest parasite mortality was found after 24 h of exposure against Haemaphysalis bispinosa (LC(50) = 404.51 ppm, r (2) = 0.890), Hippobosca maculata (LC(50) = 600.75 ppm, r (2) = 0.983), the larvae of R. microplus (LC(50) = 548.28 ppm, r (2) = 0.975), fourth-instar larvae of A. subpictus (LC(50) = 327.27 pm, r (2) = 0.970), and C. quinquefasciatus (LC(50) = 456.29 ppm, r (2) = 0.974), respectively. The control (distilled water) showed nil mortality in the concurrent assay. The χ (2) values were significant at p < 0.05 level. Therefore, the eco-friendly and biodegradable compounds from fruit peel aqueous extract of A. squamosa may be an alternative to conventional synthetic chemicals, particularly in integrated approach for the control of Haemaphysalis bispinosa, Hippobosca maculata, R. microplus, and the medically important vectors A. subpictus and C. quinquefasciatus.

Evaluation of Catharanthus roseus leaf extract-mediated biosynthesis of titanium dioxide nanoparticles against Hippobosca maculata and Bovicola ovis.

The purpose of the present study was based on assessments of the antiparasitic activities of synthesized titanium dioxide nanoparticles (TiO(2) NPs) utilizing leaf aqueous extract of Catharanthus roseus against the adults of hematophagous fly, Hippobosca maculata Leach (Diptera: Hippoboscidae), and sheep-biting louse, Bovicola ovis Schrank (Phthiraptera: Trichodectidae). The synthesized TiO(2) NPs were analyzed by X-ray diffraction (XRD), Fourier transform infrared spectroscopy (FTIR), scanning electron microscopy (SEM), and atomic force microscopy (AFM). The formation of the TiO(2) NPs synthesized from the XRD spectrum compared with the standard confirmed spectrum of titanium particles formed in the present experiments were in the form of nanocrystals, as evidenced by the peaks at 2θ values of 27.43°, 36.03°, and 54.32°. The FTIR spectra of TiO(2) NPs exhibited prominent peaks at 714 (Ti-O-O bond), 1,076 (C-N stretch aliphatic amines), 1,172 (C-O stretching vibrations in alcoholic groups), 1,642 (N-H bend bond), and 3,426 (O-H stretching due to alcoholic group). SEM analysis of the synthesized TiO(2) NPs clearly showed the clustered and irregular shapes, mostly aggregated and having the size of 25-110 nm. By Bragg's law and Scherrer's constant, it is proved that the mean size of synthesized TiO(2) NPs was 65 nm. The AFM obviously depicts the formation of the rutile and anatase forms in the TiO(2) NPs and also, the surface morphology of the particles is uneven due to the presence of some of the aggregates and individual particles. Adulticidal parasitic activity was observed in varying concentrations of aqueous leaf extract of C. roseus, TiO(2) solution, and synthesized TiO(2) NPs for 24 h. The maximum parasitic activity was observed in aqueous crude leaf extracts of C. roseus against the adults of H. maculata and B. ovis with LD(50) values of 36.17 and 30.35 mg/L, and r (2) values of 0.948 and 0.908, respectively. The highest efficacy was reported in 5 mM TiO(2) solution against H. maculata and B. ovis (LD(50) = 33.40 and 34.74 mg/L; r (2) = 0.786 and 0.873), respectively, and the maximum activity was observed in the synthesized TiO(2) NPs against H. maculata and B. ovis with LD(50) values of LD(50) = 7.09 and 6.56 mg/L, and r (2) values of 0.880 and 0.913, respectively. This method is considered as an innovative alternative approach to control the hematophagous fly and sheep-biting louse.

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.

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.

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.

Lousicidal activity of synthesized silver nanoparticles using Lawsonia inermis leaf aqueous extract against Pediculus humanus capitis and Bovicola ovis.

In the present work, we describe inexpensive, nontoxic, unreported and simple procedure for synthesis of silver nanoparticles (Ag NPs) using leaf aqueous extract of Lawsonia inermis as eco-friendly reducing and capping agent. The aim of the present study was to assess the lousicidal activity of synthesized Ag NPs against human head louse, Pediculus humanus capitis De Geer (Phthiraptera: Pediculidae), and sheep body louse, Bovicola ovis Schrank (Phthiraptera: Trichodectidae). Direct contact method was conducted to determine the potential of pediculocidal activity and impregnated method was used with slight modifications to improve practicality and efficiency of tested materials of synthesized Ag NPs against B. ovis. The synthesized Ag NPs characterized with the UV showing peak at 426 nm. X-ray diffraction (XRD) spectra clearly shows that the diffraction peaks in the pattern indexed as the silver with lattice constants. XRD analysis showed intense peaks at 2θ values of 38.34°, 44.59°, 65.04°, and 77.77° corresponding to (111), (200), (220), and (311) Bragg's reflection based on the fcc structure of Ag NPs. Fourier transform infrared spectroscopy (FTIR) spectra of Ag NPs exhibited prominent peaks at 3,422.13, 2,924.12, 2,851.76, 1,631.41, 1,381.60, 1,087.11, and 789.55 cm(-1). Scanning electron microscopy (SEM) micrograph showed mean size of 59.52 nm and aggregates of spherical shape Ag NPs. Energy dispersive X-ray spectroscopy (EDX) showed the complete chemical composition of the synthesized Ag NPs. In pediculocidal activity, the results showed that the optimal times for measuring percent mortality effects of synthesized Ag NPs were 26, 61, 84, and 100 at 5, 10, 15, and 20 min, respectively. The average percent mortality for synthesized Ag NPs was 33, 84, 91, and 100 at 10, 15, 20, and 35 min, respectively against B. ovis. The maximum activity was observed in the aqueous leaf extract of L. inermis, 1 mM AgNO(3) solution, and synthesized Ag NPs against P. humanus capitis with LC(50) values of 18.26, 7.77, and 1.33 mg l(-1) and r (2) values of 0.863, 0.900, and 0.803 and against B. ovis showed with LC(50) values of 21.19, 8.49, and 1.41 mg l(-1) and r (2) values of 0.920, 0.938 and 0.870, respectively. The findings revealed that synthesized Ag NPs possess excellent anti-lousicidal activity.

Efficacy of adulticidal and larvicidal properties of botanical extracts against Haemaphysalis bispinosa, Hippobosca maculata, and Anopheles subpictus.

The aim of the present study was to investigate the adulticidal and larvicidal activity of dried leaf hexane, ethyl acetate, acetone, and methanol extracts of Nelumbo nucifera, Manilkara zapota, Ipomoea staphylina, and Acalypha indica against the adults of Haemaphysalis bispinosa (Acarina: Ixodidae), hematophagous fly Hippobosca maculata (Diptera: Hippoboscidae), and fourth instar larvae of malaria vector Anopheles subpictus (Diptera: Culicidae). Parasites were exposed to varying concentrations of plant extracts for 24 h. All extracts showed moderate parasitic effects; however, the percent parasitic mortality observed in the crude leaf hexane, ethyl acetate, acetone, and methanol extracts of N. nucifera and M. zapota against H. bispinosa were 80, 74, 72, and 100 and 100, 83, 74, and 91, respectively, and the activity for I. staphylina and A. indica against Hip. maculata were 100, 93, 87, and 66 and 78, 90, 87, and 100 at 2,000 ppm, respectively; the larvicidal activity for the same extracts of I. staphylina against A. subpictus were 76, 82, 84, and 100 at 100 ppm, respectively. The maximum efficacy was observed in the leaf methanol extract of N. nucifera, hexane extract of M. zapota and leaf hexane extract of I. staphylina, and methanol extract of A. indica against the adults of H. bispinosa and Hip. maculata with LC(50) and LC(90) values of 437.14 and 200.81, and 415.14 and 280.72 ppm, 1,927.57 and 703.52 ppm, and 1,647.70 and 829.39 ppm, respectively. The effective larvicidal activity was observed in leaf methanol extract of I. staphylina against A. subpictus with LC(50) and LC(90) values of 10.39 and 37.71 ppm, respectively. Therefore, this study provides the first report on the adulticidal and larvicidal activity of crude solvent extracts. This is an ideal eco-friendly approach for the control of H. bispinosa, Hip. maculata, and the medically important vector A. subpictus.