Zinc oxide nanoparticles using plant Lawsonia inermis and their mosquitocidal, antimicrobial, anticancer applications showing moderate side effects.

Microbes or parasites spread vector-borne diseases by mosquitoes without being affected themselves. Insecticides used in vector control produce a substantial problem for human health. This study synthesized zinc oxide nanoparticles (ZnO NPs) using Lawsonia inermis L. and were characterized by UV-vis, FT-IR, SEM with EDX, and XRD analysis. Green synthesized ZnO NPs were highly toxic against Anopheles stephensi, whose lethal concentrations values ranged from 5.494 ppm (I instar), 6.801 ppm (II instar), 9.336 ppm (III instar), 10.736 ppm (IV instar), and 12.710 ppm (pupae) in contrast to L. inermis treatment. The predation efficiency of the teleost fish Gambusia affinis and the copepod Mesocyclops aspericornis against A. stephensi was not affected by exposure at sublethal doses of ZnO NPs. The predatory potency for G. affinis was 45 (I) and 25.83% (IV), copepod M. aspericornis was 40.66 (I) and 10.8% (IV) while in an ZnO NPs contaminated environment, the predation by the fish G. affinis was boosted to 71.33 and 34.25%, and predation of the copepod M. aspericornis was 60.35 and 16.75%, respectively. ZnO NPs inhibited the growth of several microbial pathogens including the bacteria (Escherichia coli and Bacillus subtilis) and the fungi (Alternaria alternate and Aspergillus flavus), respectively. ZnO NPs decreased the cell viability of Hep-G2 with IC50 value of 21.63 µg/mL (R2 = 0.942; P < 0.001) while the concentration increased from 1.88 to 30 µg/mL. These outcomes support the use of L. inermis mediated ZnO NPs for mosquito control and drug development.

Dietary Supplementation With Vitamins and Minerals Improves Larvae and Adult Rearing Conditions of Anopheles darlingi (Diptera: Culicidae).

Several experiments with Anopheles darlingi Root, an important malaria vector in the Amazon region, were carried out in the laboratory, depending on the large-scale production of viable larvae and adults. Certainly, improvements in rearing conditions, including dietary requirements, can strongly affect mosquito production. In order to increase the production of this species in the laboratory, we first supplemented the regular larval diet (TetraMin Tropical Flakes) with different concentrations of vitamins and minerals and recorded several biological variables: survival and larval development time, emergence ratio, and adult longevity under a small-scale rearing condition. Second, we established an experimental design under regular lab-rearing conditions based on the concentration of vitamins and minerals that best contributed to the development of these anophelines, and evaluated the biological parameters already mentioned. Moreover, under regular rearing conditions, we recorded sex ratio, adult size, and longevity of adults fed with supplemented sucrose. The lowest concentration of vitamins (V5) and the average concentration of minerals (M3) increased larval survival and decreased larval development time compared with the control. Under regular rearing conditions, minerals provided higher larval survival and increased the longevity of adults fed with supplemented sucrose. Supplementing the regular larval diet and sucrose solutions with vitamins and minerals increased the production of immatures and the longevity of An. darlingi adults.

The potential use of Azolla pinnata as an alternative bio-insecticide.

Four different tests showed the effectiveness of Azolla pinnata plant extracts against Aedes aegypti and Aedes albopictus mosquitoes. In the adulticidal test, there was a significant increase in mortality as test concentration increases and A. pinnata extracts showed LC50 and LC95 values of 2572.45 and 6100.74 ppm, respectively, against Ae. aegypti and LC50 and LC95 values of 2329.34 and 5315.86 ppm, respectively, against Ae. albopictus. The ovicidal test showed 100% eggs mortality for both species tested for all the concentrations tested at 1500 ppm, 1000 ppm, 500 ppm, 250 ppm and 125 ppm. Both tested samples of Ae. aegypti and Ae. albopictus did not lay any eggs in the plastic cups filled with the A. pinnata extract but instead opted to lay eggs in the plastic cups filled with water during the oviposition deterrence test. Similarly, the non-choice test of Ae. aegypti mosquitoes laid eggs on the sucrose solution meant for the nutrient source of the mosquitoes instead of in the plastic cup that was designed to facilitate oviposition filled with the extract. This clearly indicates the presence of bioactive compounds which are responsible in adulticidal and ovicidal activity in Aedes mosquitoes and at the same time inducing repellence towards the mosquitoes. The LC-MS results showed mainly three important chemical compounds from A. pinnata extracts such as 1-(O-alpha-D-glucopyranosyl)-(1,3R,25R)-hexacosanetriol, Pyridate and Nicotinamide N-oxide. All these chemicals have been used for various applications such as both emulsion and non-emulsion type of cosmetics, against mosquito vector such as Culex pipens and Anopheles spp. Finally, the overall view of these chemical components from A. pinnata extracts has shown the potential for developing natural product against dengue vectors.

Malaria risk map for India based on climate, ecology and geographical modelling.

Mapping the malaria risk at various geographical levels is often undertaken considering climate suitability, infection rate and/or malaria vector distribution, while the ecological factors related to topography and vegetation cover are generally neglected. The present study abides a holistic approach to risk mapping by including topographic, climatic and vegetation components into the framework of malaria risk modelling. This work attempts to delineate the areas of Plasmodium falciparum and Plasmodium vivax malaria transmission risk in India using seven geo-ecological indicators: temperature, relative humidity, rainfall, forest cover, soil, slope, altitude and the normalized difference vegetation index using multi-criteria decision analysis based on geographical information system (GIS). The weight of the risk indicators was assigned by an analytical hierarchical process with the climate suitability (temperature and humidity) data generated using fuzzy logic. Model validation was done through both primary and secondary datasets. The spatio-ecological model was based on GIS to classify the country into five zones characterized by various levels of malaria transmission risk (very high; high; moderate; low; and very low. The study found that about 13% of the country is under very high malaria risk, which includes the malaria- endemic districts of the states of Chhattisgarh, Odisha, Jharkhand, Tripura, Assam, Meghalaya and Manipur. The study also showed that the transmission risk suitability for P. vivax is higher than that for P. falciparum in the Himalayan region. The field study corroborates the identified malaria risk zones and highlights that the low to moderate risk zones are outbreak-prone. It is expected that this information will help the National Vector Borne Disease Control Programme in India to undertake improved surveillance and conduct target based interventions.

Effect of water source and feed regime on development and phenotypic quality in Anopheles gambiae (s.l.): prospects for improved mass-rearing techniques towards release programmes.

BACKGROUND: In many malaria-endemic sub-Saharan countries, insecticide resistance poses a threat to existing mosquito control measures, underscoring the need for complementary control methods such as sterile and/or genetically-modified mosquito release programmes. The sibling species Anopheles gambiae and An. coluzzii are responsible for malaria transmission in most of this region. In their natural habitat, these species generally breed in clean, soft water and it is believed that divergent preference in their larval breeding sites have played a role in their speciation process. Mosquito release programmes rely on the rearing of mosquitoes at high larval densities. Current rearing protocols often make use of deionised water regardless of the strain reared. They also depend on a delicate balance between the need for adequate feeding and the negative effect of toxic ammonia and food waste build-up on mosquito development, making managing and improving water quality in the insectary imperative. METHODS: Here, we investigated the impact of water source and feed regimes on emergence rate and phenotypic quality of mosquitoes in the insectary. First-instar larvae of An. gambiae (Kisumu strain) and An. coluzzii (Mopti and VK3 strains) were reared in three water sources with varying degrees of hardness (deionised, mineral and a mix of the two), with a daily water change. Larvae were fed daily using two standardised feeding regimes, solution and powder feed. RESULTS: Water source had a significant impact on mosquito size and development time for all strains. Earlier emergence of significantly larger mosquitoes was observed in mineral water with the smallest mosquitoes developing later from deionised water. Wing-length was significantly longer in mineral, mixed water and in powder feed, irrespective of sex, strains or water types. Deionised water was the least favourable for mosquito quality across all strains. CONCLUSIONS: Mineral water and powder feed should be used in rearing protocols to improve mosquito quality where the optimal quality of mosquitoes is desired. Although results obtained were not significant for improved mosquito numbers, the phenotypic quality of mosquitoes reared was significantly improved in mineral water and mix water. Further studies are recommended on the impact mineral water has on other fitness traits such as longevity, fecundity and mating competitiveness.

Assessment of the developmental success of Anopheles coluzzii larvae under different nutrient regimes: effects of diet quality, food amount and larval density.

BACKGROUND: In a context of increasing resistance of both vectors toward main classes of insecticides used in public health and parasites toward anti-malarial drugs, development of new and complementary molecules or control approaches is fundamental to achieve the objective of controlling or even eliminating malaria. Concerning vector control, the sterile insect technique and other genetic control approaches are among promising complementary tools in an integrated management strategy for malaria control. These approaches rely not only on a good understanding of vector biology (especially during larval stages), but also on the availability of adequate supplies and protocols for efficient mosquito rearing. The aim of this study was to assess the factors impacting the life history of Anopheles coluzzii mosquitoes at the larval stage, in the context of genetic and sterile insect approaches to control malaria vectors. METHODS: The effect of different larval diets and larval rearing densities on the development of An. coluzzii were evaluated in the laboratory. Emergence rate (ER), pre-imaginal developmental time (DT) and adult wing length (WL) were measured under different food regimes. Four diets were tested among which three were provided by the Insect Pest Control Laboratory (IPCL) of the FAO/IAEA Joint division. RESULTS: Data showed significant differences in the quality of the different diets and suggested a negative density dependence in all three life history parameters measured under tested rearing conditions. ER and WL increased with food availability, but decreased with increasing larval density. Conversely DT was shortened with increasing food availability but increased with larval density. These data demonstrates intraspecific larval competition modulated by food amount and space availability. Of the four diets tested, the one made of a mix of tuna meal, bovine liver powder, brewer's yeast, squid liver powder and vitamin mix (diet 2) yielded the best results as it produced a good balance between ER, DT and WL. Food availability for optimal development (highest survival at shortest time) was in the range of 180-400 µg/larvae/day for the three diets provided by the IPCL. CONCLUSION: There is an interaction between diet type, diet concentration and larval density. Best results in terms of optimal larvae development parameters happen when moderately high values of those three variables are observed.

Target and non-target toxicity of fern extracts against mosquito vectors and beneficial aquatic organisms.

Dengue and malaria are significant mosquito-borne diseases that are rapidly spread worldwide, mainly in temperate countries. Pteridophytes were identified to be a significant source of novel mosquitocidal agents. The present research was to explore the eco-friendly larvicides from methanol extracts of ferns, viz., Actiniopteris radiata, Adiantum caudatum, Cheilanthes swartzii, Hemionitis arifolia and Lycopodium clavatum. The larvicidal potential of the extracts screened using larvae of dengue vector Aedes aegypti (III and IV instar) and malarial vector Anopheles stephensi (III and IV instar), showed 10-100% mortality rates. Biosafety assessment was made on embryos of Danio rerio and Artemia nauplii. The phyto-constituents of the methanol extract of A. radiata leaves were identified through gas chromatography-mass spectrometry (GC-MS). Methanolic leaf extracts of A. radiata, A. caudatum and C. swartzii exhibited larvicidal activity against III and IV instar larvae of Ae. aegypti (LC50: 37.47, 74.51 and 152.38 and 67.58, 95.89 and 271.46 ppm) and An. stephensi (LC50: 70.35, 112.12 and 301.05 and 113.83, 175.30 and 315.19 ppm), respectively. The GC-MS of the methanol extract of A. radiata leaves revealed the presence of 7 phyto-components among which, Carbamic acid, phenyl-, (2-Nitrophenyl) methyl ester (1), Benzoic acid, 3- methylbenzoate (2) and 4-(benzylimino)- 1,4-dihydro-1-(p-toluoylmethyl) pyridine (3) were dominant. Biosafety assessment of methanol extract of A. radiata leaves on embryos of Danio rerio (Zebra fish) and Artemia nauplii (micro crustacean) revealed that there were no destructive or teratogenic effects. To conclude, the larvicidal activity and insignificant toxicity to non-target aquatic organisms of A. radiata leaves makes it a potential and environment safe biocontrol agent against dengue and malarial vectors.

Larvicidal activity of Zanthoxylum acanthopodium essential oil against the malaria mosquitoes, Anopheles anthropophagus and Anopheles sinensis.

BACKGROUND: Zanthoxylum acanthopodium has insecticidal effect in Chinese traditional medicine. In this study, the essential oil from the dried Zanthoxylum plant was used as a larvicidal compound against the malaria mosquitoes, Anopheles anthropophagus and Anopheles sinensis. METHODS: Compounds in the Zanthoxylum essential oil were investigated by gas chromatography and mass spectroscopy (GC-MS). The larvicidal bioassays of the whole oil, as well as the main compounds in the oil (estragole and eucalyptol) were performed using WHO method. RESULTS: In total, 63 main compounds (99.32%) were found in the oils, including estragole (15.46%), eucalyptol (10.94%), ß-caryophyllene (5.52%), cis-linalool oxide (3.76%), cis-limonene oxide (3.06%). A dose-dependent effect on mortality was recorded with increasing concentrations of essential oil and compounds increasing mortality of the larvae. Larvicidal bioassays revealed that 24 h LC50 of the whole essential oil was 36.00 mg/L and LC90 was 101.49 mg/L against An. anthropophagus, while LC50 was 49.02 mg/L and LC90 was 125.18 mg/L against An. sinensis. Additionally, 24 h LC50 of estragole were 38.56 and 41.67 mg/L against An. anthropophagus and An. sinensis, respectively, while the related LC90 were 95.90 and 107.89 mg/L. LC50 of eucalyptol were 42.41 and 45.49 mg/L against An. anthropophagus and An. sinensis, while the related LC90 were 114.45 and 124.95 mg/L. CONCLUSION: The essential oil of Z. acanthopodium and its several major compounds may have potential for use in the control of malaria mosquitoes.

Adenosine Triphosphate-Binding Cassette Transporters Are Not Involved In the Detoxification of Azadirachta indica Extracts In Anopheles stephensi Larvae.

Detoxifying pathways of mosquitoes against the neem (Azadirachta indica) extracts are still unclear. The aim of the present study was to investigate the role of adenosine triphosphate-binding cassette (ABC) transporters in this process in Anopheles stephensi, one of the main malaria vectors in southern Asia. Third-stage larvae of An. stephensi were fed with fish food alone or in combination with neem extract at 0.5%, 1%, 5%, and 10%. Six ABC-transporter genes from 3 different subfamilies (B, C, and G) were analyzed to assess their relative expression compared with controls. A bioassay was also performed to assess larval mortality rate at different concentrations and in combination with verapamil, an ABC-transporter inhibitor. No significant variation in the expression levels of any transporter belonging to the B, C, and G subfamilies was detected. Furthermore, the use of verapamil did not induce an increase in mortality at any of the tested neem extract concentrations, indicating that ABC transporters are not involved in the detoxification of neem extracts in An. stephensi larvae.

[Susceptibility status of Anopheles gambiae s.s. and An. coluzzii to Organophosphates and Carbamates in the Agricultural Exploitation Zones of oil Palm at Mouila, Gabon]. / Susceptibilité d'Anopheles gambiae s.s. et An. coluzzii aux organophosphorés et aux carbamates en zones d'exploitation industrielle de palmiers à huile à Mouila, Gabon.

This study was conducted during the rainy season of 2017 in the agricultural areas at Mouila, with the aim to assess the determination of susceptibility of Anopheles gambiae complex to insecticides and the detection of Ace 1R mutation. Mosquito larvae were collected by using the dipping method and nursed to adult stage. The susceptibility of adult Anopheles gambiae s.l. populations to organophosphates and carbamates was assessed using the standard WHO susceptibility test protocol. The tested mosquitoes species were identified by PCR. These tested mosquitoes were used to search for the Ace 1R mutation. The insecticides used were pirimiphos-methyl 0.25% and 1.25%, fenitrothion 1%, malathion 5%, propoxur 0.1%, bendiocarb 0.1%, 0.5% and 1%. In the prospected areas, An. gambiae s.l. included An. gambiae s.s. and An. coluzzii sympatric in their larval habitats. However, An. gambiae s.s. was predominant in all studied areas (99.1%) comparatively to An. coluzzii Moreover, the susceptibility tests of Anopheles to organophosphates and carbamates revealed mortality rates of 100% regardless of the origin of the mosquitoes. PCR diagnosis of Ace 1R mutation showed that none tested mosquito (An. gambiae s.s. and An. coluzzii) from prospected areas harbored the Ace 1R gene in all prospected areas. The lack of Ace 1R mutation in An. gambiae s.l. reveals the current effectiveness of organophosphates and carbamates for the control of these mosquitoes. Bboth these classes of insecticides could be used for the anopheline populations control in the agricultural spaces at Mouila.

Cette étude a été menée en 2017 pendant la saison des pluies dans les zones d'exploitations agricoles à Mouila, afin de déterminer le statut de sensibilité des membres du complexe Anopheles gambiae aux insecticides et de rechercher la présence de la mutation Ace 1R. Les moustiques ont été collectés au stade larvaire par la méthode du « dipping ¼, puis élevés jusqu'au stade adulte. La susceptibilité des adultes d'An. gambiae s.l. aux organophosphorés et aux carbamates a été évaluée par la méthode standard de l'OMS concernant les tests de sensibilité aux insecticides. Les classes d'insecticides testées étaient les organophosphorés (pirimiphos-méthyl 0,25 % et 1,25 %, fénitrothion 1 %, malathion 5 %) et les carbamates (propoxur 0,1 %, bendiocarb 0,1 %, 0,5 % et 1 %). Les moustiques testés ont été identifiés par PCR. Les anophèles issus des tests ont été utilisés pour la recherche de la mutation Ace 1R par PCR. Dans les sites d'étude, Anopheles gambiae s.l. était composé d'An. gambiae s.s. et An. coluzzii vivant en sympatrie dans leurs habitats larvaires, avec une prédominance d'An. gambiae s.s. (99,1 %) sur An. coluzzii. Les tests de sensibilité des anophèles aux organophosphorés et aux carbamates ont révélé des taux de mortalité de 100 % quelle que soit l'origine des anophèles. La PCR diagnostique de la mutation Ace 1R a montré qu'aucun moustique issu des tests de sensibilité n'était porteur de gènes de résistance Ace 1R. L'absence de mutation Ace 1R chez An. gambiae s.l. révèle l'efficacité actuelle des carbamates et des organophosphorés dans la lutte contre ces moustiques. Ces familles d'insecticides pourraient être utilisées pour la lutte contre les anophèles dans les espaces agricoles de Mouila.

Larvicidal, Ovicidal, and Histopathological Effects of the Sulphur Polypore Mushroom, Laetiporus sulphureus (Agaricomycetes), Collected from Tamil Nadu, India against Mosquitoes.

This study was conducted to determine the mosquitocidal activity of 6 species of higher Basidiomycetes collected from various places in the state of Tamil Nadu, India. The mushrooms were identified using MycoKey MMI software. We used morphological characteristics such as size, color, height, width (of the cap, lamellae, stipe, and fruiting body), spore deposits, and the presence or absence of ring to identify species. Eggs and larvae of Culex quinquefasciatus and Anopheles stephensi were exposed to various concentrations (62.5, 125.0, 250.0, and 500.0 ppm) of extracts from the 6 mushrooms. The extracts were dissolved in dimethyl sulfoxide. Among the 6 mushrooms, the methanol extract of Laetiporus sulphureus showed high toxicity toward mosquitoes at different life stages, with 96% larvicidal activity against A. stephensi and 76% activity against C. quinquefasciatus. After 24 hours, we found half-maximal lethal concentrations of 155.862 ppm for A. stephensi and 227.225 ppm for C. quinquefasciatus, and 90% lethal concentrations of 424.128 ppm for A. stephensi and 1011.663 ppm for C. quinquefasciatus. The extracts also showed strong ovicidal activity against mosquito eggs: 100% activity against A. stephensi and 91% activity against C. quinquefasciatus were observed at an extract concentration of 500 ppm 120 hours after treatment. The methanol extract of L. sulphureus was tested for toxicity against a nontarget organism, but we found none. Histopathological studies of the treated larvae revealed serious damage to cells in the midgut. These results suggest that the methanol extract of L. sulphureus is a good natural source for controlling mosquitoes.

Essential oils and their components as an alternative in the control of mosquito vectors of disease.

More than half of the human population is exposed to mosquito-borne infections. Climate change and the emergence of strains resistant to traditionally used insecticides have motivated the search of new agents for mosquito population control. Essential oils have been effective repellents and larvicidal agents.The aim of this work was to review research studies conducted in recent years on the larvicidal activity of essential oils and their components against Aedes, Anopheles and Culex mosquitoes, as well as the latest reports about their possible mechanism of action.

Grass Pollen Affects Survival and Development of Larval Anopheles arabiensis (Diptera: Culicidae).

Nutrients in breeding sites are critical for the survival and development of malaria mosquitoes, having a direct impact on vectorial capacity. Yet, there is a limited understanding about the natural larval diet and its impact on the individual fitness of mosquitoes. Recent studies have shown that gravid Anopheles arabiensis Patton (Diptera: Culicidae) are attracted by and oviposit in grass-associated habitats. The pollen provided by these grasses is a potential source of nutrients for the larvae. Here, we assess the effect of Typha latifolia L. (Poales: Typhaceae), Echinochloa pyramidalis Lamarck, Pennisetum setaceum Forsskål, and Zea mays L. pollen on larval survival and rate of development in An. arabiensis under laboratory conditions. In addition, we characterize the carbon to nitrogen ratio and the size of pollen grains as a measure of diet quality. Carbon-rich pollen with a small grain size (T. latifolia and P. setaceum; 9.7 ± 0.3 × 103 and 5.5 ± 0.2 × 104 µm3, respectively) resulted in enhanced rates of development of An. arabiensis. In contrast, the larva fed on the nitrogen-rich control diet (TetraMin) was slower to develop, but demonstrated the highest larval survival. Larvae fed on carbon-rich and large-grained Z. mays pollen (4.1 ± 0.2 × 105 µm3) survived at similar levels as those fed on the control diet and also took a longer time to develop compared with larvae fed on the other pollens. While males and females did not appear to develop differently on the different pollen diets, males consistently emerged faster than their female counterparts. These results are discussed in relation to integrated vector management.

Aceites esenciales y sus componentes como una alternativa en el control de mosquitos vectores de enfermedades / Essential oils and their components as an alternative in the control of mosquito vectors of disease

Resumen Más de la mitad de la población humana está expuesta a contraer infecciones transmitidas por mosquitos. El cambio climático y la aparición de cepas resistentes a los insecticidas tradicionalmente utilizados han motivado la búsqueda de nuevos agentes capaces de controlar las poblaciones de mosquitos. Los aceites esenciales han resultado ser eficaces agentes repelentes y larvicidas. El objetivo de este trabajo fue revisar las investigaciones llevadas a cabo en los últimos años sobre la actividad larvicida de los aceites esenciales y sus componentes contra mosquitos de los géneros Aedes, Anopheles y Culex, así como los últimos reportes sobre su posible mecanismo de acción.

Abstract More than half of the human population is exposed to mosquito-borne infections. Climate change and the emergence of strains resistant to traditionally used insecticides have motivated the search of new agents for mosquito population control. Essential oils have been effective repellents and larvicidal agents. The aim of this work was to review research studies conducted in recent years on the larvicidal activity of essential oils and their components against Aedes, Anopheles and Culex mosquitoes, as well as the latest reports about their possible mechanism of action.

Green tea proanthocyanidins cause impairment of hormone-regulated larval development and reproductive fitness via repression of juvenile hormone acid methyltransferase, insulin-like peptide and cytochrome P450 genes in Anopheles gambiae sensu stricto.

Successful optimization of plant-derived compounds into control of nuisance insects would benefit from scientifically validated targets. However, the close association between the genotypic responses and physiological toxicity effects mediated by these compounds remains underexplored. In this study, we evaluated the sublethal dose effects of proanthocyanidins (PAs) sourced from green tea (Camellia sinensis) on life history traits of Anopheles gambiae (sensu stricto) mosquitoes with an aim to unravel the probable molecular targets. Based on the induced phenotypic effects, genes selected for study targeted juvenile hormone (JH) biosynthesis, signal transduction, oxidative stress response and xenobiotic detoxification in addition to vitellogenesis in females. Our findings suggest that chronic exposure of larval stages (L3/L4) to sublethal dose of 5 ppm dramatically extended larval developmental period for up to 12 days, slowed down pupation rates, induced abnormal larval-pupal intermediates and caused 100% inhibition of adult emergence. Further, females exhibited significant interference of fecundity and egg hatchability relative to controls (p < 0.001). Using reverse transcription quantitative polymerase chain reaction (RT-qPCR), our findings show that PA-treated larvae exhibited significant repression of AgamJHAMT (p < 0.001), AgamILP1 (p < 0.001) and AgamCYP6M2 (p < 0.001) with up-regulation of Hsp70 (p < 0.001). Females exposed as larvae demonstrated down-regulation of AgamVg (p = 0.03), AgamILP1 (p = 0.009), AgamCYP6M2 (p = 0.05) and AgamJHAMT (p = 0.02). Our findings support that C. sinensis proanthocyanidins affect important vectorial capacity components such as mosquito survival rates and reproductive fitness thus could be potentially used for controlling populations of malaria vectors.

Larvicidal activity of phytosteroid compounds from leaf extract of Solanum nigrum against Culex vishnui group and Anopheles subpictus.

BACKGROUND: Vector control is facing a menace due to the appearance of resistance to synthetic insecticides. Insecticides of plant origin may provide appropriate substitute biocontrol techniques in the future. The present study was carried out to investigate the bio control potentiality of active ingredient isolated from chloroform: methanol (1:1 v/v) extract of mature leaves of Solanum nigrum L. (Solanaceae) against early 3rd instar larvae of Culex vishnui group (comprising of Cx. vishnui Theobald, Cx. pseudovishnui Colless and Cx. tritaeniorhynchus Giles) and Anopheles subpictus Grassi. S. nigrum is a common plant distributed in many parts of India with medicinal properties. METHODS: Bioactive compound isolated from chloroform: methanol (1:1 v/v) extract of mature leaves of S. nigrum was (25, 45, 60 mg/L) tested against early 3rd instar larvae of Cx. vishnui group and An. subpictus. The lethal concentration was determined by log probit analysis. The chemical nature of the active substance was also evaluated following gas chromatography-mass spectroscopy (GC-MS) and infrared (IR) analysis. The compound was also studied on non target organisms such as Daphnia sp. and Diplonychus annulatum. RESULTS: TLC spot having Rf value of 0.94 (Rf = 14.1/15 = 0.94) showed larvicidal activity. In a 72 h bioassay experiment, mortality rate at 60 mg/L was significantly higher (P < 0.05) than those at 25 and 45 mg/L against early 3rd instar. Result of log-probit analysis (at 95% confidence level) revealed that LC50 and LC90 values gradually decreased with the exposure period showing the lowest value at 72 h of exposure. A clear dose-dependent mortality was observed, as the rate of mortality (Y) was positively correlated with the concentration (X) having regression coefficient value close to one in each case. The compound was found to be eco-friendly as it did not show any adverse effect to the studied non target organisms. Chemical characterization (GC-MS and IR analyses) of the active ingredient revealed the presence of phytosteroid compounds responsible for mosquito larvicidal activity. CONCLUSION: Leaf extract of S. nigrum has great potential as bio control agent against Cx. vishnui group and An. subpictus. In near future the isolated bioactive phytochemical could be used as a source of an effective mosquitocidal agent.

Larvicidal activity of Blumea eriantha essential oil and its components against six mosquito species, including Zika virus vectors: the promising potential of (4E,6Z)-allo-ocimene, carvotanacetone and dodecyl acetate.

The effective and environmentally sustainable control of mosquitoes is a challenge of essential importance. This is due to the fact that some invasive mosquitoes, with special reference to the Aedes genus, are particularly difficult to control, due to their high ecological plasticity. Moreover, the indiscriminate overuse of synthetic insecticides resulted in undesirable effects on human health and non-target organisms, as well as resistance development in targeted vectors. Here, the leaf essential oil (EO) extracted from a scarcely studied plant of ethno-medicinal interest, Blumea eriantha (Asteraceae), was tested on the larvae of six mosquitoes, including Zika virus vectors. The B. eriantha EO was analyzed by GC and GC-MS. The B. eriantha EO showed high toxicity against 3rd instar larvae of six important mosquito species: Anopheles stephensi (LC50=41.61 µg/ml), Aedes aegypti (LC50=44.82 µg/ml), Culex quinquefasciatus (LC50 =48.92 µg/ml), Anopheles subpictus (LC50=51.21 µg/ml), Ae. albopictus (LC50=56.33 µg/ml) and Culex tritaeniorhynchus (LC50=61.33 µg/ml). The major components found in B. eriantha EO were (4E,6Z)-allo-ocimene (12.8%), carvotanacetone (10.6%), and dodecyl acetate (8.9%). Interestingly, two of the main EO components, (4E,6Z)-allo-ocimene and carvotanacetone, achieved LC50 lower than 10 µg/ml on all tested mosquito species. The acute toxicity of B. eriantha EO and its major constituents on four aquatic predators of mosquito larval instars was limited, with LC50 ranging from 519 to 11.431 µg/ml. Overall, the larvicidal activity of (4E,6Z)-allo-ocimene and carvotanacetone far exceed most of the LC50 calculated in current literature on mosquito botanical larvicides, allowing us to propose both of them as potentially alternatives for developing eco-friendly mosquito control tools.

Toxicity of ar-curcumene and epi-ß-bisabolol from Hedychium larsenii (Zingiberaceae) essential oil on malaria, chikungunya and St. Louis encephalitis mosquito vectors.

Mosquitoes act as vectors of key pathogens and parasites. Plant essential oils have been recognized as important sources of biopesticides, which do not induce resistance and have limited toxic effects on human health and non-target organisms. In this research, we evaluated the larvicidal and oviposition deterrence activity of Hedychium larsenii essential oil (EO) and its major compounds ar-curcumene and epi-ß-bisabolol. Both molecules showed high toxicity against early third instars of Anopheles stephensi (LC50=10.45 and 14.68µg/ml), Aedes aegypti (LC50=11.24 and 15.83µg/ml) and Culex quinquefasciatus (LC50=12.24 and 17.27µg/ml). In addition, low doses of ar-curcumene and epi-ß-bisabolol were effective as oviposition deterrents against the three tested mosquito species. Notably, the acute toxicity of H. larsenii oil and its major compounds against the mosquito biocontrol agent Poecilia reticulata was low, with LC50 higher than 1500ppm. Overall, the results from this study revealed that ar-curcumene and epi-ß-bisabolol from the H. larsenii oil can be considered for the development of novel and effective mosquito larvicides.

Methanolic extract of Agerantum conyzoides exhibited toxicity and growth disruption activities against Anopheles gambiae sensu stricto and Anopheles arabiensis larvae.

BACKGROUND: Vector control remains the mainstay to effective malaria management. The negative implications following persistent application of synthetic insecticides geared towards regulation of mosquito populations have necessitated prospection for ecofriendly effective chemistries. Plant-derived compounds have the potential to control malaria-transmitting mosquito populations. Previously, Agerantum conyzoides extracts have demonstrated toxicity effects on disease-transmitting mosquitoes. However, their efficacy in controlling Afrotropical malaria vectors remains unclear. Herein, the toxicity and growth disruption activities of crude methanolic leaf extract of A. conyzoides on Anopheles gambiae sensu stricto and An. arabiensis larvae were assessed. METHODS: Late third (L3) instars of An. gambiae s.s and An. arabiensis larvae were challenged with increasing doses of crude methanolic extract of A. conyzoides. The larval mortality rates were recorded every 24 h and the LC50 values determined at their associated 95% confidence levels. ANOVA followed by Post-hoc Student-Newman-Keuls (SNK) test was used to compare results between treatment and control groups. Phytochemical profiling of the extract was performed using standard chemical procedures. RESULTS: Treatment of larvae with the methanolic extract depicted dose-dependent effects with highest mortality percentages of ≥ 69% observed when exposed with 250 ppm and 500 ppm for 48 h while growth disruption effects were induced by sublethal doses of between 50-100 ppm for both species. Relative to experimental controls, the extract significantly reduced larval survival in both mosquito species (ANOVA, F(8,126) = 43.16776, P < 0.001). The LC50 values of the extract against An. gambiae s.s ranged between 84.71-232.70 ppm (95% CI 81.17-239.20), while against An. arabiensis the values ranged between 133.46-406.35 ppm (95% CI 131.51-411.25). The development of the juvenile stages was arrested at pupal-larval intermediates and adult emergence. The presence of alkaloids, aglycone flavonoids, triterpenoids, tannins and coumarins can partly be associated with the observed effects. CONCLUSION: The extract displayed considerable larvicidal activity and inhibited emergence of adult mosquitoes relative to experimental controls, a phenomenon probably associated with induced developmental hormone imbalance. Optimization of the bioactive compounds could open pathways into vector control programmes for improved mosquito control and reduced malaria transmission rates.

Traditional herbal remedies and dietary spices from Cameroon as novel sources of larvicides against filariasis mosquitoes?

In Cameroon, many dietary spices are used by traditional healers to cure several diseases such as cancer and microbial infections. Aframomum daniellii, Dichrostachys cinerea and Echinops giganteus are Cameroonian spices widely used as flavourings and as food additives. Moreover, they are traditionally herbal remedies employed to treat several diseases, as well as to control populations of insect pests. In this research, we analysed the chemical composition of A. daniellii, D. cinerea and E. giganteus essential oils and we evaluated their larvicidal potential against larvae of the filariasis and West Nile virus vector Culex quinquefasciatus. The essential oils were obtained from different plant parts by hydrodistillation and their composition was analysed by GC-MS. The three spices exhibited different volatile chemical profiles, being characterized by 1,8-cineole, sabinene and ß-pinene (A. daniellii), geraniol and terpinen-4-ol (D. cinerea), and silphiperfol-6-ene and presilphiperfolan-8-ol (E. giganteus). Results showed that the highest larvicidal toxicity on Cx. quinquefasciatus was exerted by D. cinerea essential oil (LC50 = 39.1 µL L-1), followed by A. daniellii (pericarp essential oil: LC50 = 65.5 µL L-1; leaves: LC50 = 65.5µL L-1; seeds: LC50 = 106.5µL L-1) and E. giganteus (LC50 = 227.4 µL L-1). Overall, the chance to use the D. cinerea essential oil against Cx. quinquefasciatus young instars seems promising, since it is effective at moderate doses and could be an advantageous alternative to build newer mosquito control tools.