Malaria Vector Anopheles Stephensi Research Articles

One-step synthesis of polydispersed silver nanocrystals using Malva sylvestris: an eco-friendly mosquito larvicide with negligible impact on non-target aquatic organisms.

The synthesis of eco-friendly nanoparticles is evergreen branch of nanoscience with a growing number of biomedical implications. In this study, we investigated the synthesis of polydisperse and stable silver nanoparticles (AgNP) using a cheap leaf extract of Malva sylvestris (Malvaceae). Bio-reduced AgNP were characterized by UV-visible spectrophotometry, Fourier transform infrared spectroscopy (FTIR), X-ray diffraction analysis (XRD), atomic force microscopy (AFM), scanning electron microscopy (SEM) and transmission electron microscopy (TEM). The acute toxicity of M. sylvestris leaf extract and green-synthesized AgNP was evaluated against larvae of the malaria vector Anopheles stephensi, the dengue vector Aedes aegypti and the filariasis vector Culex quinquefasciatus. Compared to the leaf aqueous extract, AgNP showed higher toxicity against A. stephensi, A. aegypti, and C. quinquefasciatus with LC50 values of 10.33, 11.23, and 12.19μg/mL, respectively. Green-synthesized AgNP were found safer to non-target organisms Diplonychus indicus and Gambusia affinis, with respective LC50 values ranging from 813.16 to 1044.52μg/mL. Overall, this research firstly shed light on the mosquitocidal potential of M. sylvestris, a potential bio-resource for rapid, cheap and effective synthesis of polydisperse and highly stable silver nanocrystals.

Chemical composition, toxicity and non-target effects of Pinus kesiya essential oil: An eco-friendly and novel larvicide against malaria, dengue and lymphatic filariasis mosquito vectors

Mosquitoes (Diptera: Culicidae) are vectors of important parasites and pathogens causing death, poverty and social disability worldwide, with special reference to tropical and subtropical countries. The overuse of synthetic insecticides to control mosquito vectors lead to resistance, adverse environmental effects and high operational costs. Therefore, the development of eco-friendly control tools is an important public health challenge. In this study, the mosquito larvicidal activity of Pinus kesiya leaf essential oil (EO) was evaluated against the malaria vector Anopheles stephensi, the dengue vector Aedes aegypti and the lymphatic filariasis vector Culex quinquefasciatus. The chemical composition of the EO was analyzed by gas chromatography–mass spectroscopy. GC–MS revealed that the P. kesiya EO contained 18 compounds. Major constituents were α-pinene, β-pinene, myrcene and germacrene D. In acute toxicity assays, the EO showed significant toxicity against early third-stage larvae of An. stephensi, Ae. aegypti and Cx. quinquefasciatus, with LC50 values of 52, 57, and 62µg/ml, respectively. Notably, the EO was safer towards several aquatic non-target organisms Anisops bouvieri, Diplonychus indicus and Gambusia affinis, with LC50 values ranging from 4135 to 8390µg/ml. Overall, this research adds basic knowledge to develop newer and safer natural larvicides from Pinaceae plants against malaria, dengue and filariasis mosquito vectors.

Genome Sequence of Stenotrophomonas maltophilia Strain SmAs1, Isolated From the Asian Malaria Mosquito Anopheles stephensi.

An isolate of Stenotrophomonas maltophilia was cultured from the Asian malaria vector Anopheles stephensi. Here, we present the annotated draft genome sequence of this S. maltophilia strain. This genomic resource will facilitate further characterization of bacteria associated with mosquitoes.

Characterization and expression analysis of gene encoding heme peroxidase HPX15 in major Indian malaria vector Anopheles stephensi (Diptera: Culicidae)

The interaction of mosquito immune system with Plasmodium is critical in determining the vector competence. Thus, blocking the crucial mosquito molecules that regulate parasite development might be effective in controlling the disease transmission. In this study, we characterized a full-length AsHPX15 gene from the major Indian malaria vector Anopheles stephensi. This gene is true ortholog of Anopheles gambiae heme peroxidase AgHPX15 (AGAP013327), which modulates midgut immunity and regulates Plasmodium falciparum development. We found that AsHPX15 is highly induced in mosquito developmental stages and blood fed midguts. In addition, this is a lineage-specific gene that has identical features and 65–99% amino acids identity with other HPX15 genes present in eighteen worldwide-distributed anophelines. We discuss that the conserved HPX15 gene might serve as a common target to manipulate mosquito immunity and arresting Plasmodium development inside the vector host.

Mosquito repellent potential of Pithecellobium dulce leaf and seed against malaria vector Anopheles stephensi (Diptera: Culicidae)

Objective: To determine the repellent properties of hexane, benzene, ethyl acetate, chloroform and methanol extract of Pithecellobium dulce (P. dulce) leaf and seed against Anopheles stephensi (An. stephensi). Methods: Repellent activity assay was carried out in a net cage (45 cm 伊 30 cm 伊 25 cm) containing 100 blood starved female mosquitoes of An. stephensi. This assay was carried out in the laboratory conditions according to the WHO 2009 protocol. Plant crude extracts of P. dulce were applied at 1.0, 2.5, and 5.0 mg/cm 2 separately in the exposed fore arm of study subjects. Ethanol was used as the sole control. Results: In this study, the applied plant crude extracts were observed to protect against mosquito bites. There were no allergic reactions experienced by the study subjects. The repellent activity of the extract was dependent on the concentration of the extract. Among the tested solvents, the leaf and seed methanol extract showed the maximum efficacy. The highest concentration of 5.0 mg/cm 2 leaf and seed methanol extract of P. dulce provided over 180 min and 150 min protection, respectively. Conclusions: Crude extracts of P. dulce exhibit the potential for controlling malaria vector mosquito An. stephensi. Journal homepage: www.jclmm.com

Larvicidal and repellent potential of Zingiber nimmonii (J. Graham) Dalzell (Zingiberaceae) essential oil: an eco-friendly tool against malaria, dengue, and lymphatic filariasis mosquito vectors?

Mosquitoes (Diptera: Culicidae) are important vectors of terms of public health relevance, especially in tropical and sub-tropical regions. The continuous and indiscriminate use of conventional pesticides for the control of mosquito vectors has resulted in the development of resistance and negative impacts on non-target organisms and the environment. Therefore, there is a need for development of effective mosquito control tools. In this study, the larvicidal and repellent activity of Zingiber nimmonii rhizome essential oil (EO) was evaluated against the malaria vector Anopheles stephensi, the dengue vector Aedes aegypti, and the lymphatic filariasis vector Culex quinquefasciatus. The chemical composition of the EO was analyzed by gas chromatography-mass spectroscopy (GC-MS). GC-MS revealed that the Z. nimmonii EO contained at least 33 compounds. Major constituents were myrcene, β-caryophyllene, α-humulene, and α-cadinol. In acute toxicity assays, the EO showed significant toxicity against early third-stage larvae of An. stephensi, Ae. aegypti, and Cx. quinquefasciatus, with LC50 values of 41.19, 44.46, and 48.26μg/ml, respectively. Repellency bioassays at 1.0, 2.0, and 5.0mg/cm(2) of Z. nimmonii EO gave 100% protection up to 120, 150, and 180min. against An. stephensi, followed by Ae. aegypti (90, 120, and 150min) and Cx. quinquefasciatus (60, 90, and 120min). Furthermore, the EO was safer towards two non-target aquatic organisms, Diplonychus indicus and Gambusia affinis, with LC50 values of 3241.53 and 9250.12μg/ml, respectively. Overall, this research adds basic knowledge to develop newer and safer natural larvicides and repellent from Zingiberaceae plants against malaria, dengue, and filariasis mosquito vectors.

A yeast strain associated to Anopheles mosquitoes produces a toxin able to kill malaria parasites.

BackgroundMalaria control strategies are focusing on new approaches, such as the symbiotic control, which consists in the use of microbial symbionts to prevent parasite development in the mosquito gut and to block the transmission of the infection to humans. Several microbes, bacteria and fungi, have been proposed for malaria or other mosquito-borne diseases control strategies. Among these, the yeast Wickerhamomyces anomalus has been recently isolated from the gut of Anopheles mosquitoes, where it releases a natural antimicrobial toxin. Interestingly, many environmental strains of W. anomalus exert a wide anti-bacterial/fungal activity and some of these ‘killer’ yeasts are already used in industrial applications as food and feed bio-preservation agents. Since a few studies showed that W. anomalus killer strains have antimicrobial effects also against protozoan parasites, the possible anti-plasmodial activity of the yeast was investigated.MethodsA yeast killer toxin (KT), purified through combined chromatographic techniques from a W. anomalus strain isolated from the malaria vector Anopheles stephensi, was tested as an effector molecule to target the sporogonic stages of the rodent malaria parasite Plasmodium berghei, in vitro. Giemsa staining was used to detect morphological damages in zygotes/ookinetes after treatment with the KT. Furthermore, the possible mechanism of action of the KT was investigated pre-incubating the protein with castanospermine, an inhibitor of β-glucanase activity.ResultsA strong anti-plasmodial effect was observed when the P. berghei sporogonic stages were treated with KT, obtaining an inhibition percentage up to around 90 %. Microscopy analysis revealed several ookinete alterations at morphological and structural level, suggesting the direct implication of the KT-enzymatic activity. Moreover, evidences of the reduction of KT activity upon treatment with castanospermine propose a β-glucanase-mediated activity.ConclusionThe results showed the in vitro killing efficacy of a protein produced by a mosquito strain of W. anomalus against malaria parasites. Further studies are required to test the KT activity against the sporogonic stages in vivo, nevertheless this work opens new perspectives for the possible use of killer strains in innovative strategies to impede the development of the malaria parasite in mosquito vectors by the means of microbial symbionts.Electronic supplementary materialThe online version of this article (doi:10.1186/s12936-015-1059-7) contains supplementary material, which is available to authorized users.

Lectin-carbohydrate recognition mechanism of Plasmodium berghei in the midgut of malaria vector Anopheles stephensi using quantum dot as a new approach

Potential targets of Plasmodium ookinetes at the mosquito midgut walls were investigated in relation to interfering malarial transmission. In this study, the essential application of Quantum Dots (QDs) was used to examine the interaction between Plasmodium berghei ookinetes and the Anopheles stephensi midgut, based on lectin-carbohydrate recognition. Two significant lectins were utilized to determine this interaction.Two QDs, cadmium telluride (CdTe)/CdS and cadmium selenide (CdSe)/CdS, were employed in staining Plasmodium ookinete to study its interaction in the midgut of the mosquito vector in vivo. Concurrently, two lectins, wheat germ agglutinin (WGA) and concanavalin A (Con A), were inadvertently exploited to mask lectin binding sites between ookinetes and mosquito midgut cells. The numbers of ookinetes in both lumen and epithelial cells were eventually counted, following adequate preparation of wax sections extracted from whole midgut, and subsequent examination using a differential interference contrast a fluorescence microscopic technique.Interestingly, we detected that neither of the QDs mutated ookinete invasion into the midgut cells of the investigated mosquitoes. QD staining of ookinetes remained permanent despite the effective embedding procedure. The massive binding potency of ookinetes to midgut cells of the cross-examined mosquitoes undoubtedly revealed that Con A did not interrupt ookinete penetration into the midgut wall. In contrast, WGA inhibited ookinete invasion into the midgut cells. The results proved that QD nanoparticles are biocompatible, non-toxic to P. berghei and stable to photobleaching. The QDs staining, which was successfully implemented for ookinete labelling, is a simple and effective tool which plays a crucial role in bioimaging including the study of parasite-vector interactions.

Larvicidal potential of carvacrol and terpinen-4-ol from the essential oil of Origanum vulgare (Lamiaceae) against Anopheles stephensi, Anopheles subpictus, Culex quinquefasciatus and Culex tritaeniorhynchus (Diptera: Culicidae)

Mosquito-borne diseases represent a deadly threat for millions of people worldwide. However, the use of synthetic insecticides to control Culicidae may lead to resistance, high operational costs and adverse non-target effects. Nowadays, plant-borne mosquitocides may serve as suitable alternative in the fight against mosquito vectors. In this study, the mosquito larvicidal activity of Origanum vulgare (Lamiaceae) leaf essential oil (EO) and its major chemical constituents was evaluated against the malaria vectors Anopheles stephensi and An. subpictus, the filariasis vector Culex quinquefasciatus and the Japanese encephalitis vector Cx. tritaeniorhynchus. The chemical composition of the EO was analyzed by gas chromatography–mass spectroscopy. GC–MS revealed that the essential oil of O. vulgare contained 17 compounds. The major chemical components were carvacrol (38.30%) and terpinen-4-ol (28.70%). EO had a significant toxic effect against early third-stage larvae of An. stephensi, An. subpictus, Cx. quinquefasciatus and Cx. tritaeniorhynchus, with LC50 values of 67.00, 74.14, 80.35 and 84.93μg/ml. The two major constituents extracted from the O. vulgare EO were tested individually for acute toxicity against larvae of the four mosquito vectors. Carvacrol and terpinen-4-ol appeared to be most effective against An. stephensi (LC50=21.15 and 43.27μg/ml, respectively) followed by An. subpictus (LC50=24.06 and 47.73μg/ml), Cx. quinquefasciatus (LC50=26.08 and 52.19μg/ml) and Cx. tritaeniorhynchus (LC50=27.95 and 54.87μg/ml). Overall, this research adds knowledge to develop newer and safer natural larvicides against malaria, filariasis and Japanese encephalitis mosquito vectors.

Highly efficient Cas9-mediated gene drive for population modification of the malaria vector mosquito Anopheles stephensi

Genetic engineering technologies can be used both to create transgenic mosquitoes carrying antipathogen effector genes targeting human malaria parasites and to generate gene-drive systems capable of introgressing the genes throughout wild vector populations. We developed a highly effective autonomous Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)-associated protein 9 (Cas9)-mediated gene-drive system in the Asian malaria vector Anopheles stephensi, adapted from the mutagenic chain reaction (MCR). This specific system results in progeny of males and females derived from transgenic males exhibiting a high frequency of germ-line gene conversion consistent with homology-directed repair (HDR). This system copies an ∼ 17-kb construct from its site of insertion to its homologous chromosome in a faithful, site-specific manner. Dual anti-Plasmodium falciparum effector genes, a marker gene, and the autonomous gene-drive components are introgressed into ∼ 99.5% of the progeny following outcrosses of transgenic lines to wild-type mosquitoes. The effector genes remain transcriptionally inducible upon blood feeding. In contrast to the efficient conversion in individuals expressing Cas9 only in the germ line, males and females derived from transgenic females, which are expected to have drive component molecules in the egg, produce progeny with a high frequency of mutations in the targeted genome sequence, resulting in near-Mendelian inheritance ratios of the transgene. Such mutant alleles result presumably from nonhomologous end-joining (NHEJ) events before the segregation of somatic and germ-line lineages early in development. These data support the design of this system to be active strictly within the germ line. Strains based on this technology could sustain control and elimination as part of the malaria eradication agenda.

Plant-mediated biosynthesis of nanoparticles as an emerging tool against mosquitoes of medical and veterinary importance: a review.

Mosquitoes (Diptera: Culicidae) are a key threat for millions of people worldwide, since they act as vectors for devastating parasites and pathogens. Mosquito young instars are usually targeted with organophosphates, insect growth regulators and microbial control agents. Indoors residual spraying and insecticide-treated bed nets are also employed. However, these chemicals have strong negative effects on human health and the environment. Newer and safer tools have been recently implemented to enhance control of mosquitoes. In this review, I focus on characterization, effectiveness, and non-target effects of mosquitocidal nanoparticles synthesized using botanical products (mosquitocidal nanoparticles, MNP). The majority of plant-fabricated MNP are silver ones. The synthesis of MNP is usually confirmed by UV-visualization spectroscopy, followed by scanning electron microscopy or transmission electron microscopy, energy-dispersive X-ray spectroscopy, Fourier transform infrared spectroscopy, and X-ray diffraction studies. Interestingly, plant-synthesized metal nanoparticles have been reported as effective ovicides, larvicides, pupicides, adulticides, and oviposition deterrents against different mosquito species of medical and veterinary importance. Few parts per million of different MNP are highly toxic against the malaria vector Anopheles stephensi, the dengue vector Aedes aegypti, and the filariasis mosquito Culex quiquefasciatus. However, despite the growing number of evidences about the effectiveness of MNP, moderate efforts have been carried out to shed light on their possible non-target effects against mosquito's natural enemies and other aquatic organisms. In the final section, particular attention was dedicated to this issue. A number of hot areas that need further research and cooperation among parasitologists and entomologists are highlighted.

Phospholipid Topography of Whole-Body Sections of the Anopheles stephensi Mosquito, Characterized by High-Resolution Atmospheric-Pressure Scanning Microprobe Matrix-Assisted Laser Desorption/Ionization Mass Spectrometry Imaging.

High-resolution atmospheric-pressure scanning microprobe matrix-assisted laser desorption/ionization mass spectrometry imaging (AP-SMALDI MSI) has been employed to study the molecular anatomical structure of rodent malaria vector Anopheles stephensi mosquitoes. A dedicated sample preparation method was developed which suits both, the special tissue properties of the sample and the requirements of high-resolution MALDI imaging. Embedding in 5% carboxymethylcellulose (CMC) was used to maintain the tissue integrity of the whole mosquitoes, being very soft, fragile, and difficult to handle. Individual lipid compounds, specifically representing certain cell types, tissue areas, or organs, were detected and imaged in 20 μm-thick whole-body tissue sections at a spatial resolution of 12 μm per image pixel. Mass spectrometric data and information quality were based on a mass resolution of 70,000 (at m/z 200) and a mass accuracy of better than 2 ppm in positive-ion mode on an orbital trapping mass spectrometer. A total of 67 imaged lipids were assigned by database search and, in a number of cases, identified via additional MS/MS fragmentation studies directly from tissue. This is the first MSI study at 12 μm spatial resolution of the malaria vector Anopheles. The study provides insights into the molecular anatomy of Anopheles stephensi and the distribution and localization of major classes of glycerophospholipids and sphingolipids. These data can be a basis for future experiments, investigating, e.g., the metabolism of Plasmodium-infected and -uninfected Anopheles mosquitoes.

Eco-friendly control of malaria and arbovirus vectors using the mosquitofish Gambusia affinis and ultra-low dosages of Mimusops elengi-synthesized silver nanoparticles: towards an integrative approach?

Mosquito-borne diseases represent a deadly threat for millions of people worldwide. However, the use of synthetic insecticides to control Culicidae may lead to high operational costs and adverse non-target effects. Plant-borne compounds have been proposed for rapid extracellular synthesis of mosquitocidal nanoparticles. Their impact against biological control agents of mosquito larval populations has been poorly studied. We synthesized silver nanoparticles (AgNP) using the aqueous leaf extract of Mimusops elengi as a reducing and stabilizing agent. The formation of AgNP was studied using different biophysical methods, including UV-vis spectrophotometry, TEM, XRD, EDX and FTIR. Low doses of AgNP showed larvicidal and pupicidal toxicity against the malaria vector Anopheles stephensi and the arbovirus vector Aedes albopictus. AgNP LC50 against A. stephensi ranged from 12.53 (I instar larvae) to 23.55ppm (pupae); LC50 against A. albopictus ranged from 11.72ppm (I) to 21.46ppm (pupae). In the field, the application of M. elengi extract and AgNP (10 × LC50) led to 100% larval reduction after 72h. In adulticidal experiments, AgNP showed LC50 of 13.7ppm for A. stephensi and 14.7ppm for A. albopictus. The predation efficiency of Gambusia affinis against A. stephensi and A. albopictus III instar larvae was 86.2 and 81.7%, respectively. In AgNP-contaminated environments, predation was 93.7 and 88.6%, respectively. This research demonstrates that M. elengi-synthesized AgNP may be employed at ultra-low doses to reduce larval populations of malaria and arbovirus vectors, without detrimental effects on predation rates of mosquito natural enemies, such as larvivorous fishes.

Seaweed-synthesized silver nanoparticles: an eco-friendly tool in the fight against Plasmodium falciparum and its vector Anopheles stephensi?

Malaria, the most widespread mosquito-borne disease, affects 350-500 million people each year. Eco-friendly control tools against malaria vectors are urgently needed. This research proposed a novel method of plant-mediated synthesis of silver nanoparticles (AgNP) using a cheap seaweed extract of Ulva lactuca, acting as a reducing and capping agent. AgNP were characterized by UV-vis spectrophotometry, Fourier transform infrared (FTIR) spectroscopy, energy-dispersive X-ray spectroscopy (EDX), scanning electron microscopy (SEM), and X-ray diffraction (XRD). The U. lactuca extract and the green-synthesized AgNP were tested against larvae and pupae of the malaria vector Anopheles stephensi. In mosquitocidal assays, LC50 values of U. lactuca extract against A. stephensi larvae and pupae were 18.365 ppm (I instar), 23.948 ppm (II), 29.701 ppm (III), 37.517 ppm (IV), and 43.012 ppm (pupae). LC50 values of AgNP against A. stephensi were 2.111 ppm (I), 3.090 ppm (II), 4.629 ppm (III), 5.261 ppm (IV), and 6.860 ppm (pupae). Smoke toxicity experiments conducted against mosquito adults showed that U. lactuca coils evoked mortality rates comparable to the permethrin-based positive control (66, 51, and 41%, respectively). Furthermore, the antiplasmodial activity of U. lactuca extract and U. lactuca-synthesized AgNP was evaluated against CQ-resistant (CQ-r) and CQ-sensitive (CQ-s) strains of Plasmodium falciparum. Fifty percent inhibitory concentration (IC50) values of U. lactuca were 57.26 μg/ml (CQ-s) and 66.36 μg/ml (CQ-r); U. lactuca-synthesized AgNP IC50 values were 76.33 μg/ml (CQ-s) and 79.13 μg/ml (CQ-r). Overall, our results highlighted out that U. lactuca-synthesized AgNP may be employed to develop newer and safer agents for malaria control.

Mosquitocidal and antiplasmodial activity of Senna occidentalis (Cassiae) and Ocimum basilicum (Lamiaceae) from Maruthamalai hills against Anopheles stephensi and Plasmodium falciparum.

Each year, mosquito-borne diseases infect nearly 700 million people, resulting to more than 1 million deaths. In this study, we evaluated the larvicidal, pupicidal, and smoke toxicity of Senna occidentalis and Ocimum basilicum leaf extracts against the malaria vector Anopheles stephensi. Furthermore, the antiplasmodial activity of plant extracts was evaluated against chloroquine (CQ)-resistant (CQ-r) and CQ-sensitive (CQ-s) strains of Plasmodium falciparum. In larvicidal and pupicidal experiments, S. occidentalis LC50 ranged from 31.05 (I instar larvae) to 75.15 ppm (pupae), and O. basilicum LC50 ranged from 29.69 (I instar larvae) to 69 ppm (pupae). Smoke toxicity experiments conducted against adults showed that S. occidentalis and O. basilicum coils evoked mortality rates comparable to the pyrethrin-based positive control (38, 52, and 42%, respectively). In antiplasmodial assays, Senna occidentalis 50% inhibitory concentration (IC50) were 48.80 μg/ml (CQ-s) and 54.28 μg/ml (CQ-r), while O. basilicum IC50 were 68.14 μg/ml (CQ-s) and 67.27 μg/ml (CQ-r). Overall, these botanicals could be considered as potential sources of metabolites to build newer and safer malaria control tools.

Bioefficacy of Morinda tinctoria and Pongamia glabra plant extracts against the malaria vector Anopheles stephensi (Diptera: Culicidae)

Mosquito-borne diseases have an economic impact, including loss in commercial and labour outputs, particularly in countries with tropical and subtropical climates; however, no part of the world is free from vector-borne diseases. The aim of the present study was to investigate the larvicidal, adulticidal and ovicidal activity of dried leaf chloroform, ethyl acetate, acetone, aqueous, and methanol extracts of <em>Morinda</em> <em>tinctoria</em> and <em>Pongamia</em> <em>glabra</em> against larvae of <em>Anopheles</em> <em>stephensi</em> (Diptera: Culicidae). Larvae were exposed to varying concentrations of plant extracts for 24 h. All extracts showed moderate larvicidal effects after 24 h of exposure; however, the highest larval mortality was found with the leaf methanol extracts of <em>M. tinctoria</em> and <em>P. glabra</em> against the larvae of <em>A. stephensi</em> lethal concentration (LC)<sub>50</sub>=136.24 and 141.05 ppm; LC<sub>90</sub>=342.67 and 368.89 ppm, respectively. The results of the adulticidal activity assays of chloroform, ethyl acetate, acetone, aqueous, and methanol extracts of <em>M. tinctoria</em> and <em>P. glabra</em> showed significant mortality against larvae of <em>A. stephensi</em>. The methanol extract showed maximum activity compared with the other extracts. The greatest effect on mean percentage hatch in the ovicidal assays was observed 48 h post-treatment. Percent hatch was inversely proportional to the concentration of extract, and directly proportional to the number of eggs. A mortality of 100% was observed with 100-400 ppm methanol extracts and 200-400 ppm aqueous extracts of <em>M. tinctoria</em>, and 200-400 ppm aqueous and methanol extracts of <em>P. glabra</em>. This study provides the first report of the larvicidal, adulticidal and ovicidal activities of <em>M. tinctoria</em> and <em>P. glabra</em> plant extracts against the malaria vector, A. stephensi, representing an ideal eco-friendly approach for its control.

Cymbopogon citratus-synthesized gold nanoparticles boost the predation efficiency of copepod Mesocyclops aspericornis against malaria and dengue mosquitoes

Plant-borne compounds can be employed to synthesize mosquitocidal nanoparticles that are effective at low doses. However, how they affect the activity of mosquito predators in the aquatic environment is unknown. In this study, we synthesized gold nanoparticles (AuN) using the leaf extract of Cymbopogon citratus, which acted as a reducing and capping agent. AuN were characterized by a variety of biophysical methods and sorted for size in order to confirm structural integrity. C. citratus extract and biosynthesized AuN were tested against larvae and pupae of the malaria vector Anopheles stephensi and the dengue vector Aedes aegypti. LC50 of C. citratus extract ranged from 219.32 ppm to 471.36 ppm. LC50 of AuN ranged from 18.80 ppm to 41.52 ppm. In laboratory, the predatory efficiency of the cyclopoid crustacean Mesocyclops aspericornis against A. stephensi larvae was 26.8% (larva I) and 17% (larva II), while against A. aegypti was56% (I) and 35.1% (II). Predation against late-instar larvae was minimal. In AuN-contaminated environment,predation efficiency against A. stephensi was 45.6% (I) and 26.7% (II), while against A. aegypti was 77.3% (I) and 51.6% (II). Overall, low doses of AuN may help to boost the control of Anopheles and Aedes larval populations in copepod-based control programs.

Mosquitocidal and antibacterial activity of green-synthesized silver nanoparticles from Aloe vera extracts: towards an effective tool against the malaria vector Anopheles stephensi?

Mosquitoes represent an important threat for lives of millions of people worldwide, acting as vectors for devastating pathogens, such as malaria, yellow fever, dengue, and West Nile. In addition, pathogens and parasites polluting water also constitute a severe plague for populations of developing countries. Here, we investigated the mosquitocidal and antibacterial properties of Aloe vera leaf extract and silver nanoparticles synthesized using A. vera extract. Mosquitocidal properties were assessed in laboratory against larvae (I-IV instar) and pupae of the malaria vector Anopheles stephensi. Green-synthesized silver nanoparticles were tested against An. stephensi also in field conditions. Antibacterial properties of nanoparticles were evaluated against Bacillus subtilis, Klebsiella pneumoniae, and Salmonella typhi using the agar disk diffusion and minimum inhibitory concentration protocol. The synthesized silver nanoparticles were characterized by UV-vis spectrum, Fourier transform infrared spectroscopy (FTIR), scanning electron microscopy (SEM), and X-ray diffraction (XRD). In laboratory conditions, the A. vera extract was toxic against An. stephensi larvae and pupae, even at low dosages. LC50 were 48.79 ppm (I instar), 59.09 ppm (II instar), 70.88 ppm (III instar), 83.58 ppm (IV instar), and 152.55 ppm (pupae). Green-synthesized silver nanoparticles were highly toxic against An. stephensi. LC50 were 3.825 ppm (I instar), 4.119 ppm (II instar), 4.982 ppm (III instar), 5.711 ppm (IV instar), and 6.113 ppm (pupae). In field conditions, the application of A. vera-synthesized silver nanoparticles (10 × LC50) leads to An. stephensi larval reduction of 74.5, 86.6, and 97.7%, after 24, 48, and 72 h, respectively. Nanoparticles also showed antibacterial properties, and the maximum concentration tested (150 mg/L) evoked an inhibition zone wider than 80 mm in all tested bacterium species. This study adds knowledge about the use of green synthesis of nanoparticles in medical entomology and parasitology, allowing us to propose A. vera-synthesized silver nanoparticles as effective candidates to develop newer and safer mosquitocidal control tools.

Biosynthesized silver nanoparticles using floral extract of Chrysanthemum indicum L.--potential for malaria vector control.

Mosquitoes transmit serious human diseases, causing millions of deaths every year. The use of synthetic insecticides to control vector mosquitoes has caused physiological resistance and adverse environmental effects in addition to high operational cost. Insecticides synthesized of natural products for vector control have been a priority in this area. In the present study, silver nanoparticles (Ag NPs) were green-synthesized using a floral extract of Chrysanthemum indicum screened for larvicidal and pupicidal activity against the first to fourth instar larvae and pupae of the malaria vector Anopheles stephensi mosquitoes. The synthesized Ag NPs were characterized by using UV-vis absorption, X-ray diffraction, transmission electron microscopy, and energy-dispersive X-ray spectroscopy techniques. The textures of the yielded Ag NPs were found to be spherical and polydispersed with a mean size in the range of 25-59nm. Larvae and pupae were exposed to various concentrations of aqueous extract of C. indicum and synthesized Ag NPs for 24h, and the maximum mortality was observed from the synthesized Ag NPs against the vector A. stephensi (LC50 = 5.07, 10.35, 14.19, 22.81, and 35.05ppm; LC90 = 29.18, 47.15, 65.53, 87.96, and 115.05ppm). These results suggest that the synthesized Ag NPs have the potential to be used as an ideal eco-friendly approach for the control of A. stephensi. Additionally, this study provides the larvicidal and pupicidal properties of green-synthesized Ag NPs with the floral extract of C. indicum against vector mosquito species from the geographical location of India.

Impact of repeated NeemAzal-treated blood meals on the fitness of Anopheles stephensi mosquitoes.

BackgroundHerbal remedies are widely used in many malaria endemic countries to treat patients, in particular in the absence of anti-malarial drugs and in some settings to prevent the disease. Herbal medicines may be specifically designed for prophylaxis and/or for blocking malaria transmission to benefit both, the individual consumer and the community at large. Neem represents a good candidate for this purpose due to its inhibitory effects on the parasite stages that cause the clinical manifestations of malaria and on those responsible for infection in the vector. Furthermore, neem secondary metabolites have been shown to interfere with various physiological processes in insect vectors. This study was undertaken to assess the impact of the standardised neem extract NeemAzal® on the fitness of the malaria vector Anopheles stephensi following repeated exposure to the product through consecutive blood meals on treated mice.MethodsBatches of An. stephensi mosquitoes were offered 5 consecutive blood meals on female BALB/c mice treated with NeemAzal® at an azadirachtin A concentration of 60, 105 or 150 mg/kg. The blood feeding capacity was estimated by measuring the haematin content of the rectal fluid excreted by the mosquitoes during feeding. The number of eggs laid was estimated by image analysis and their hatchability assessed by direct observations.ResultsA dose and frequency dependent impact of NeemAzal® treatment on the mosquito feeding capacity, oviposition and egg hatchability was demonstrated. In the 150 mg/kg treatment group, the mosquito feeding capacity was reduced by 50% already at the second blood meal and by 50 to 80% in all treatment groups at the fifth blood meal. Consequently, a 50 – 65% reduction in the number of eggs laid per female mosquito was observed after the fifth blood meal in all treatment groups. Similarly, after the fifth treated blood meal exposure, hatchability was found to be reduced by 62% and 70% in the 105 and 150 mg/kg group respectively.ConclusionsThe findings of this study, taken together with the accumulated knowledge on neem open the challenging prospects of designing neem-based formulations as multi-target phytomedicines exhibiting preventive, parasite transmission-blocking as well as anti-vectorial properties.