Duration and dose-dependency of female sexual receptivity responses to seminal fluid proteins in Aedes albopictus and Ae. aegypti mosquitoes

Male mosquitoes transfer seminal fluid proteins (hereafter ‘SFPs’) during mating. These proteins can have profound effects on female behavior in the yellow fever mosquito Aedes aegypti and the Asian tiger mosquito Aedes albopictus. SFPs are thought to be responsible for female refractoriness to mating in both species. However, only limited information is available about the duration of induced refractoriness or the quantity of SFPs required to be effective in Ae. albopictus. Here, we tested the duration of the effect of SFPs on female refractory behavior for both Aedes species. Additionally, we determined the lowest SFP dose required to induce female refractory behavior in Ae. aegypti. Virgin females were injected intra-thoracically with doses ranging from 0.25 to 0.008 equivalents of one male’s SFP amount. Our results demonstrate high sensitivity of female Ae. aegypti and Ae. albopictus to SFPs of their own species, with the majority of females becoming refractory at doses ? 0.031 male-equivalents after injection into the hemocoel. This effect was long-lasting in both species; none of the injected females were inseminated when presented with males of their own species 30 to 34 days post-injection, whereas most saline-injected control females mated at this time point. These results will aid future work to characterize individual SFPs involved in post-mating refractoriness in these two species. Moreover, they show that as is the situation in the mosquito Anopheles gambiae, and unlike Drosophila melanogaster, sperm are not required for the maintenance of a sexual refractoriness response in Ae. aegypti and Ae. albopictus.     

Toxicity of Acalypha indica (Euphorbiaceae) and Achyranthes aspera (Amaranthaceae) leaf extracts to Aedes aegypti (Diptera: Culicidae)

Alternative control strategies for the dengue vector Aedes aegypti L. (Diptera: Culicidae) include botanical insecticides. They are believed to pose little threat to the environment or to human health and may provide practical substitutes for synthetic insecticides. In this study, we determined the biological activities of methanol extracts of Acalypha indica L. (Euphorbiaceae) and Achyranthes aspera L (Amaranthaceae) leaves individually and in combination as botanical insecticides against Ae. aegypti. Based on LC₅₀ values for 4th instar Ae. aegypti, the combined extracts showed the strongest larvicidal activity (277 ppm). A. aspera and A. indica extracts individually gave similar results (409 and 420 ppm, respectively). Respective LC₅₀ values for pupae were 326 ppm, 456 ppm, and 467 ppm. In studies of smoke toxicity, 64% of females exposed to negative control smoke (no extract) blood fed on chicken, whereas 17% blood fed when exposed to smoke containing A. aspera extract and to positive control smoke (0.2% d-allethrin). In the field, treatment of water storage tanks (≈ 0.5 m³) with combined plant extract reduced larval and pupal populations by 97% and 81%, respectively, after 5 days. Given the results of this study, further evaluation of the combined (A. indica + A. aspera) extract as a mosquito larvicide is warranted. Mosquito coils with A. aspera extract also show promise as a practical and potentially economical means for mitigating mosquito blood feeding.     

Effects of AeDNV infection on Aedes aegypti lifespan and reproduction

The effects of Aedes Densovirus (AeDNV) infections on survival, fertility, fecundity and vertical transmission in Aedes aegypti (Diptera: Culicidae) were measured in laboratories in Kiev, Ukraine and Colorado, USA and incorporated into a predictive model of the effects of AeDNV on vector capacity. Adult lifespan and daily survival were reduced in AeDNV infected mosquitoes. This effect was dependent on the dose of the virus. Infected females had decreased fecundity. The oviposition rate was less in infected females and the hatch rate declined in eggs laid by infected females. The amounts of AeDNV in infected females and the infection rate of their offspring were measured with real-time PCR. The average filial transmission rate was ∼70% and larval infection rates from infected females varied between 42 and 62%. Vertically infected larvae, and individual eggs contained ∼1 × 10⁵ AeDNV genome equivalents (geq). Modeling the effects of AeDNV infection on Ae. aegypti populations suggested a large decrease in the numbers of eggs, larvae, pupae, and adults arising from infected mothers and suggested that AeDNV treatment of larvae could cause up to a 76% reduction of infectious mosquito days.     

Chrysanthemum extract and extract prepared silver nanoparticles as biocides to control Aedes aegypti (L.), the vector of dengue fever

Mosquitoes play a key role in the transmission of some important diseases. The need for controlling these insects is critical to reduce their risks to human and domesticated animals. Recently the trend to explore effective chemical compounds from local plants has begun as a safe means of control. The present study aimed to evaluate the anti-larval activity of Chrysanthemum extract and the prepared silver nanoparticle (AgNPs) against the Aedes aegypti mosquito, the dengue vector in Saudi Arabia. A series of different concentrations of ethanol extract and extract prepared AgNPs against the fourth-life larvae was tested. The effective concentrations of crude extract and AgNPs ranged from 50 to 250 and 10 to 30 ppm respectively, and the death percentages corresponding to these concentrations ranged from 18 to 92 and 36 to 96% respectively. According to the LC₅₀ values of treated larvae, AgNPs (12.754 ppm) is more effective against A. aegypti mosquito larvae than the crude extract (228.345 ppm) at about 17.9 times. The mixing of the plant extract with the silver nitrate has led to potentiation. This is due to the synergy that occurs between the extract and the silver particles during the reduction process. The compounds in the extract are related to the surface of the particles, increasing the strength of their effects. It is recommend to separate the active elements in the Chrysanthemum plant and its preparation in the form of nanoparticles as a promising compound in mosquito control programs with least damage to human kind and the environment.     

Potential for integrated control of Culex quinquefasciatus (Diptera: Culicidae) using larvicides and guppies

Acute toxicity tests were carried out to determine the effect of three larvicides, spinosad, pirimiphos methyl, and chlorpyrifos, on Culex quinquefasciatus Say, Anopheles gambiae s.s. Giles (Diptera: Culicidae), and guppies, Poecilia reticulata Peters (Pisces: Poeciliidae). Thereafter, larvae of Cx. quinquefasciatus were introduced to P. reticulata in containers of different volumes with low concentrations of each larvicide at established predator to prey densities of 1–35; 5–70 and 10–350 (fish to larvae) respectively. The experiment was replicated six times, and the larval consumption was counted after 24 h. Spinosad and pirimiphos methyl were significantly toxic to Cx. quinquefasciatus, the less susceptible mosquito species. Guppy consumption of Culex larvae was highest at a predator to prey density 5–70. Feeding activity of guppies increased in the spinosad treatment at 49 μg L⁻¹ compared to control and the synthetic larvicides. The synthetic larvicides generally reduced consumption of larvae except for the lowest concentration of pirimiphos methyl which increased it at the highest predator to prey density in a 3 L container. The highest percentage of Culex larvae was consumed by fish in the presence of spinosad at 49 μg L⁻¹ and a predator to prey density of 5–70 in 0.5 L plastic containers. Thus, predator to prey density, container size, type and concentration of larvicide are all important factors to be considered in integrated management of mosquito larvae.     

Mosquito larvicidal activity of Aloe vera (Family: Liliaceae) leaf extract and Bacillus sphaericus,against Chikungunya vector,Aedes aegypti

The bio-efficacy of Aloe vera leaf extract and bacterial insecticide, Bacillus sphaericus larvicidal activity was assessed against the first to fourth instars larvae of Aedes aegypti, under the laboratory conditions. The plant material was shade dried at room temperature and powdered coarsely. A. vera and B. sphaericus show varied degrees of larvicidal activity against various instars larvae of A. aegypti. The LC₅₀ of A. vera against the first to fourth instars larvae were 162.74, 201.43, 253.30 and 300.05 ppm and the LC₉₀ 442.98, 518.86, 563.18 and 612.96 ppm, respectively. B. sphaericus against the first to fourth instars larvae the LC₅₀ values were 68.21, 79.13, 93.48, and 107.05 ppm and the LC₉₀ values 149.15, 164.67, 183.84, and 201.09 ppm, respectively. However, the combined treatment of A. vera + B. sphaericus (1:2) material shows highest larvicidal activity of the LC₅₀ values 54.80, 63.11, 74.66 and 95.10 ppm; The LC₉₀ values of 145.29, 160.14, 179.74 and 209.98 ppm, against A. aegypti in all the tested concentrations than the individuals and clearly established that there is a substantial amount of synergist act. The present investigation clearly exhibits that both A. vera and B. sphaericus materials could serve as a potential larvicidal agent. Since, A. aegypti is a container breeder vector mosquito this user and eco-friendly and low-cost vector control strategy could be a viable solution to the existing dengue disease burden. Therefore, this study provides first report on the mosquito larvicidal activity the combined effect of A. vera leaf extract and B. sphaericus against as target species of A. aegypti.     

Productivity of natural and artificial containers for Aedes polynesiensis and Aedes aegypti in four American Samoan villages

Six mosquito species were identified in a survey of containers associated with 347 households in four villages in American Samoa. Aedes polynesiensis Marks (Diptera: Culicidae) and Aedes aegypti (L) were the most abundant species, representing 57% and 29% of the mosquitoes identified. Culex quinquefasciatus (Say), Culex annulirostris (Skuse), Aedes oceanicus (Belkin) and Toxorhynchites amboinensis (Doleschall) were also found. Aedes aegypti and Ae. polynesiensis showed distinct differences in their use of containers, preferring large and small containers, respectively. By contrast with previous studies, Ae. polynesiensis utilized domestic and natural containers with equal frequency, whereas Ae. aegypti continued to be found predominantly in domestic containers. Only 15% of containers holding immature mosquitoes included pupae and fewer than 10 Aedes spp. pupae were found in most containers with pupae. An estimated 2289 Ae. polynesiensis and 1640 Ae. aegypti pupae were found in 2258 containers. The presence of both species in the same container did not affect the mean density of either species for larvae or pupae. Glass jars, leaf axils, tree holes and seashells produced few Aedes spp. pupae in any of the study villages. Overall, 75% of Ae. polynesiensis pupae were found in buckets, ice-cream containers and tyres, with <7% being produced in natural containers, whereas 82% of Ae. aegypti pupae were found in 44-gallon (US) drums ( approximately 166L), buckets and tyres. Source reduction efforts targeting these container types may yield significant reductions in both Ae. polynesiensis and Ae. aegypti populations in American Samoa.     

Bioassays for comparative infectivity of mermithid nematodes (Romanomermis iyengari,Romanomermis culicivorax and Strelkovimermis spiculatus) for culicine mosquito larvae

Two improved bioassays were developed to establish infectivity baselines for selection experiments using mermithid nematode variants. Comparative infectivity of Romanomermis iyengari, Romanomermis culicivorax and Strelkovimermis spiculatus using larvae of three mosquito spp. Aedes sierrensis, Aedes aegypti and Culex pipiens were evaluated with “plate” and “tray” bioassays at selected intensity of infections. Using the “plate” bioassay, single mosquito larvae were immersed in 2 ml of water within individual depressions of 12-well, polystyrene tissue culture plates. One, three, or five preparasitic juveniles (J2) were added to each well. In the “tray” bioassay, polyethylene trays containing 500 ml water and 100 mosquito larvae were exposed to 500 (5:1, nematode:insect host) or 1000 (10:1) J2s. Percentage infection (PINF, infectivity) and intensity of infection (IINF, #nematodes per infected larvae) number were determined only after emergence of post-parasitic J3 juveniles. Under the bioassay conditions, all three species of nematodes resulted in infections in all mosquito hosts, but R. iyengari exhibited better effectiveness in the parasitism of mosquito larvae. The three species of mosquitoes presented high levels of susceptibility to each of the three species of nematodes, but in general Cx. pipiens and Ae. sierrensis were slightly more susceptible than Ae. aegypti. The “plate” bioassay was more efficient in measurement of infectivity of the mermithid species and in establishing baseline characteristics for these mosquito-parasitic nematodes. The “tray” bioassay was an effective bioassay for large cohorts of both infective juveniles and host larvae and, potential for field interactions.     

Larvicidal and mosquito repellent action of peppermint (Mentha piperita) oil

Oil of Mentha piperita L. (Peppermint oil), a widely used essential oil, was evaluated for larvicidal activity against different mosquito species: Aedes aegypti, Anopheles stephensi and Culex quinquefasciatus by exposing IIIrd instar larvae of mosquitoes in enamel trays 6 × 4 inch² size filled to a depth of 3 inch with water. Of the three species tested Cx. quinquefasciatus was most susceptible followed by Ae. aegypti and An. stephensi. Application of oil at 3 ml/m² of water surface area resulted in 100% mortality within 24 h for Cx. quinquefasciatus, 90% for Ae. aegypti and 85% for An. stephensi. For Ae. aegypti 100% mortality was achieved at 3 ml/m² in 48 h or 4 ml/m² in 24 h. For An. stephensi 100% mortality was observed at 4 ml/m² in 72 h. The emergence at 3 ml/m² was also inhibited to a great extent and the few adults which emerged did not ovipost even after taking a blood meal. The oil showed strong repellent action against adult mosquitoes when applied on human skin. Percent protection obtained against An. annularis, An. culicifacies, and Cx. quinquefasciatus was 100%, 92.3% and 84.5%, respectively. The repellent action of Mentha oil was comparable to that of Mylol oil consisting of dibutyl and dimethyl phthalates.     

Prevalence and resting behaviour of dengue vectors,Aedes aegypti and Aedes albopictus in dengue high risk urban settings in Colombo,Sri Lanka

Understanding local geographical variation in vector density and bionomics related to virus transmission are critical for planning effective vector control programs to control dengue virus transmission. This study investigated the prevalence and resting behaviour of Aedes aegypti and Ae. albopictus in three dengue high-risk areas in the Colombo District, Sri Lanka. Monthly sampling of resting adult mosquitoes was conducted from August 2019 to February 2020. Thirty-seven percent of the households (289/776) harboured Aedes mosquitoes, and 603 Aedes mosquitoes were collected. The proportion of Ae. aegypti was higher in the overall collection during the collection period (94% [569/603]), and 62% (352/569) were females. Significant monthly variations in Ae. aegypti were observed with respect to the indices: number of females per surveyed house (F/SH; p = 0.001), number of females per Aedes positive house (F/PH; p = 0.029), adult house index (AHI; p = 0.001), adult density (AD; p = 0.005) and resting ratio (RR; p = 0.001). AHI, AD, and RR had statistically significant positive correlations with monthly rainfall (p = 0.001, p = 0.011, and p = 0.002 respectively) and one-month lagged dengue cases (p = 0.002, p = 0.005, and p < 0.001 respectively). A statistically significantly higher proportion of Ae. aegypti females were caught resting indoors (N = 309, 88%) than outdoors (N = 43, 12%; p < 0.001). The most common resting areas were bedrooms (51%) & living/dining rooms (37%), and places were under or on furniture (47%) & hangings (34%) for Ae. aegypti. Conversely, 74% of female Ae. albopictus were collected outdoors. Results of this study could have strong implications to improve vector surveillance and control by early detection of dengue to detect outbreaks and minimization of disease transmission.     

Further study on Ascogregarina culicis in temperate Argentina: Prevalence and intensity in Aedes aegypti larvae and pupae

The bionomics of South American strains of Ascogregarina spp. are poorly known and the first studies were performed a few years ago. Our main objective was to characterize Ascogregarina culicis population in Aedes aegypti immatures in temperate Argentina. A total of 1800 water-filled containers were inspected within a cemetery of Buenos Aires City through a reproductive period of the host (October 2006-June 2007). The parasite was detected in 16.7% (329/1974) of the immatures and 8.5% (15/177) of the breeding sites. The prevalence decreased from 19.9% in larvae to 6.5% in pupae. In those infected breeding sites, about 85% of the immature mosquitoes harbor the parasite with a median intensity of nine trophozoites per larva and six gametocysts per pupa. The prevalence in shaded containers was higher than in sun exposed ones but the intensity of the infection was quite similar between both lighting conditions. Sun-exposed containers recorded water temperatures significantly higher than those under shade throughout the study period. Parasite trophozoites were only found from January to May with a clear seasonal pattern of prevalence. Monthly values of parasite prevalence and mosquito host (percentage of breeding sites and number of immatures) were significantly correlated at p < 0.05 when a temporal delay of two months was considered. Our results suggest that parasite prevalence is spatially and temporally heterogeneous in temperate urban Argentina, and these variations are associated with the host abundance.     

Comparative study of three herbal formulations against dengue vectors Aedes aegypti

The efficacy of three formulations (i.e., natural lavender crude, essential oil, and gel) extracted from Lavender angustifolia was tested against vectors of the epidemic dengue virus, Aedesaegypti, to evaluate their larvicidal activity effect. The ethanolic extract of the lavender crude was prepared using a rotary evaporator, while the other extracts, such as essential oil and gel, were obtained from iHerb, a supplier of medicinal herbs in the US. The mortality rate of larvae was evaluated 24 h after exposure. Larvicidal activity of the lavender crude was 91% mortality at 150 ppm, 94% for essential oil at a concentration of 3000 ppm, and 97% for lavender gel at a 1000 ppm. Natural lavender crude was one of the most promising extracts tested against Ae.aegypti larvae, with lethal concentrations at LC₅₀ and LC₉₀ of 76.4 and 174.5 ppm post-treatment. The essential oil had the least effect on mosquito larvae, with LC₅₀ and LC₉₀ reaching 1814.8 and 3381.9 ppm, respectively. The lavender gel was moderately effective against Ae. aegypti larvae, with LC₅₀ and LC₉₀ values reaching 416.3 and 987.7 ppm after exposure. The occurrence of morphological abnormalities in the larvae treated with the three compounds, in turn, resulted in an incomplete life cycle. Therefore, our results indicated that natural lavender crude displayed the highest larvicidal activity against larvae, followed by gel and essential oil. Thus, this study concluded that lavender crude is an effective, eco-friendly compound that can be used as an alternative to chemical products to control vector-borne epidemic diseases.     

Ongoing challenges in the management of malaria

Background

Mosquitoes transmit serious human diseases, causing millions of deaths every year. Use of synthetic insecticides to control vector mosquitoes has caused physiological resistance and adverse environmental effects in addition to high operational cost. Insecticides of botanical origin have been reported as useful for control of mosquitoes. Azadirachta indica (Meliaceae) and its derived products have shown a variety of insecticidal properties. The present paper discusses the larvicidal activity of neem-based biopesticide for the control of mosquitoes.

Methods

Larvicidal efficacy of an emulsified concentrate of neem oil formulation (neem oil with polyoxyethylene ether, sorbitan dioleate and epichlorohydrin) developed by BMR & Company, Pune, India, was evaluated against late 3^rd and early 4^th instar larvae of different genera of mosquitoes. The larvae were exposed to different concentrations (0.5–5.0 ppm) of the formulation along with untreated control. Larvicidal activity of the formulation was also evaluated in field against Anopheles, Culex, and Aedes mosquitoes. The formulation was diluted with equal volumes of water and applied @ 140 mg a.i./m² to different mosquito breeding sites with the help of pre calibrated knapsack sprayer. Larval density was determined at pre and post application of the formulation using a standard dipper.

Results

Median lethal concentration (LC₅₀) of the formulation against Anopheles stephensi, Culex quinquefasciatus and Aedes aegypti was found to be 1.6, 1.8 and 1.7 ppm respectively. LC₅₀ values of the formulation stored at 26°C, 40°C and 45°C for 48 hours against Ae. aegypti were 1.7, 1.7, 1.8 ppm while LC₉₀ values were 3.7, 3.7 and 3.8 ppm respectively. Further no significant difference in LC₅₀ and LC₉₀ values of the formulation was observed against Ae. aegypti during 18 months storage period at room temperature. An application of the formulation at the rate of 140 mg a.i./m² in different breeding sites under natural field conditions provided 98.1% reduction of Anopheles larvae on day 1; thereafter 100% reduction was recorded up to week 1 and more than 80% reduction up to week 3, while percent reduction against Culex larvae was 95.5% on day 1, and thereafter 80% reduction was achieved up to week 3. The formulation also showed 95.1% and, 99.7% reduction of Aedes larvae on day 1 and day 2 respectively; thereafter 100% larval control was observed up to day 7.

Conclusion

The neem oil formulation was found effective in controlling mosquito larvae in different breeding sites under natural field conditions. As neem trees are widely distributed in India, their formulations may prove to be an effective and eco-friendly larvicide, which could be used as an alternative for malaria control.     

Winter Refuge for Aedes aegypti and Ae. albopictus Mosquitoes in Hanoi during Winter

Dengue occurs throughout the year in Hanoi, Vietnam, despite winter low temperatures <10°C. During July 2010 to March 2012, we surveyed monthly for Aedes larvae and pupae in 120 houses in 8 Hanoi districts. Aedes albopictus preferred discarded containers in summer and pupal density drastically decreased in winter. Aedes aegypti preferred concrete tanks and this preference increased in winter. Even in winter, the lowest water temperature found in concrete tanks was >14°C, exceeding the developmental zero point of Ae. aegypti. Although jars, drums and concrete tanks were the dominant containers previously (1994–97) in Hanoi, currently the percentage of residences with concrete tanks was still high while jars and drums were quite low. Our study showed that concrete tanks with broken lids allowing mosquitoes access were important winter refuge for Ae. aegypti. We also indicate a concern about concrete tanks serving as foci for Ae. aegypti to expand their distribution in cooler regions.     

Immature mosquitoes in agricultural wetlands of the coastal plain of Georgia,U.S.A.: Effects of landscape and environmental habitat characteristics

This study is the first to report on the relationships between immature mosquitoes (larvae and pupae) and landscape and environmental habitat characteristics in wetlands associated with row crop agriculture. Indicator species analysis (ISA) was used to test for associations among mosquito species and groups of wetland sites with similar Landscape Development Intensity (LDI) values. Results indicated that Anopheles quadrimaculatus, Culex erraticus, and Psorophora columbiae were associated with agricultural wetlands (LDI > 2.0), whereas Anopheles crucians and Culex territans were associated with forested reference wetlands (LDI < 2.0) in both wet and dry years. The species fidelity to wetland type, regardless of the hydrologic regime, demonstrates these species are robust indicators of wetland condition. Data on immature mosquito assemblages were compared to selected landscape and environmental habitat variables using Akaike's Information Criterion (AIC_c) model selection. LDI indices, dissolved oxygen concentration, the proportion of emergent vegetation, and the proportion of bare ground in wetlands were important factors associated with the selected mosquito species. These results indicate that LDI indices are useful in predicting the distributions of disease vectors or other nuisance mosquito species across broad geographic areas. Additionally, these results suggest mosquitoes are valuable bioindicators of wetland condition that reflect land use and hydrologic variability.     

Chicory (Cichorium intybus) and wormwood (Artemisia absinthium) extracts exhibit strong larvicidal activity against mosquito vectors of malaria,dengue fever,and filariasis

Vector-borne diseases transmitted by mosquitoes cause globally important diseases such as malaria, dengue fever, and filariasis. The incidence of these diseases can be reduced through mosquito control programs but these control programs currently rely on synthetic insecticides that can impact the environment, and has selected widespread mosquito resistance. Environment friendly and biodegradable natural insecticides discovered in plants offer an alternative approach to mosquito control. Here, we investigated extracts from root or aerial parts of Chicory (Cichorium intybus) and wormwood (Artemisia absinthium) against the early 4th instar larvae of Anopheles stephensi (malaria vector), Aedes aegypti (dengue fever vector), and Culex quinquefasciatus (filariasis vector). The root and aerial parts extracts of A. absinthium and C. intybus at 200, 100, 50, 25 and 12.5?ppm caused significant mortality of the tested mosquito species. Root extracts exhibited higher larvicidal activity that aerial part extracts. The highest larvicidal activity was recorded in methanol extract of roots of C. intybus with LC50?=?66.16, 18.88 and LC¬90?=?197.56, 107.16?ppm for An. stephensi; LC50?=?78.51, 40.15 and LC90?=?277.31, 231.28?ppm for Ae. aegypti and LC50?=?103.99, 64.56 and LC¬90?=?314.04, 247.54?ppm for Cx. quinquefasciatus. These results reveal potent mosquito larvicidal activity against vectors of malaria, dengue fever, and filariasis is present in extracts of chicory and wormwood.     

Earthworm-mediated synthesis of silver nanoparticles: A potent tool against hepatocellular carcinoma,Plasmodium falciparum parasites and malaria mosquitoes

The development of parasites and pathogens resistant to synthetic drugs highlighted the needing of novel, eco-friendly and effective control approaches. Recently, metal nanoparticles have been proposed as highly effective tools towards cancer cells and Plasmodium parasites. In this study, we synthesized silver nanoparticles (EW–AgNP) using Eudrilus eugeniae earthworms as reducing and stabilizing agents. EW–AgNP showed plasmon resonance reduction in UV–vis spectrophotometry, the functional groups involved in the reduction were studied by FTIR spectroscopy, while particle size and shape was analyzed by FESEM. The effect of EW–AgNP on in vitro HepG2 cell proliferation was measured using MTT assays. Apoptosis assessed by flow cytometry showed diminished endurance of HepG2 cells and cytotoxicity in a dose-dependent manner. EW–AgNP were toxic to Anopheles stephensi larvae and pupae, LC₅₀ were 4.8 ppm (I), 5.8 ppm (II), 6.9 ppm (III), 8.5 ppm (IV), and 15.5 ppm (pupae). The antiplasmodial activity of EW–AgNP was evaluated against CQ-resistant (CQ-r) and CQ-sensitive (CQ-s) strains of Plasmodium falciparum. EW–AgNP IC₅₀ were 49.3 μg/ml (CQ-s) and 55.5 μg/ml (CQ-r), while chloroquine IC₅₀ were 81.5 μg/ml (CQ-s) and 86.5 μg/ml (CQ-r). EW–AgNP showed a valuable antibiotic potential against important pathogenic bacteria and fungi. Concerning non-target effects of EW–AgNP against mosquito natural enemies, the predation efficiency of the mosquitofish Gambusia affinis towards the II and II instar larvae of A. stephensi was 68.50% (II) and 47.00% (III), respectively. In EW–AgNP-contaminated environments, predation was boosted to 89.25% (II) and 70.75% (III), respectively. Overall, this research highlighted the EW–AgNP potential against hepatocellular carcinoma, Plasmodium parasites and mosquito vectors, with little detrimental effects on mosquito natural enemies.     

Effects of high-temperature stress and heat shock on two root maggots, Bradysia odoriphaga and Bradysia difformis (Diptera: Sciaridae)

Bradysia odoriphaga and B. difformis (Diptera: Sciaridae) are devastating pests of vegetables, ornamentals and edible mushrooms. In Chinese chive fields, the two Bradysia species occur with similar regularities: outbreaks in spring and autumn, and population decreases in summer. Temperature may be an important factor restricting their population abundance in summer. Here, we performed a life-table study under constant high temperatures and assessed the tolerance of two Bradysia species to heat shock. Life parameters of the Bradysia species indicated slow developmental rates, and low survival rates and fecundity, when the temperature was higher than 30 °C. At 34 °C, individuals were unable to reach the adult stages from eggs. Moreover, temperatures above 36 °C showed lethal effects, decreasing their survival rates. The median lethal time (LT50) values of 4th instar B. odoriphaga and B. difformis larvae were 46.82 and 32.97 h, respectively, while the values at 38 °C were 2.12 and 1.51 h, respectively. The 4th instar larvae and pupae possessed higher thermotolerance levels than adults and eggs, indicating sensitivities to heat stress. Moreover, B. odoriphaga was more thermotolerant than B. difformis. Thus, weak thermotolerance levels may restrict their occurrences during the period of summer heat, and the difference in thermotolerance levels between the two species may be related to their regional distributions.     

Consequences of the introduction of the planarian Girardia anceps (Tricladida: Dugesiidae) in artificial containers with larvae of the mosquitoes Aedes aegypti and Culex pipiens (Diptera: Culicidae) from Argentina

Aedes aegypti and Culex pipiens are container-dwelling mosquito species that are vectors of important diseases to man, such as dengue and lymphatic filariasis, respectively. Predators of these pests are an interesting alternative to be incorporated to biological control measures. We tested the consequences of introducing individuals of Girardia anceps, a native freshwater flatworm species, within artificial water containers where larvae of these mosquitoes thrive. Our goals were to ascertain if mosquito species, density of larvae (high or low), type of water container (tires or ovitraps), and presence or absence of planarians affected mosquito survivorship (measured as number of individuals reaching the pupa stage) in manipulated artificial containers. Furthermore, we monitored ovitraps in the field along several months in order to explore the long-term effect of the presence of planarian on the colonization of these containers by feral mosquitoes under natural conditions. We found that the presence of planarians reduced the number of mosquitoes reaching pupation and that such reduction depends on the initial density of larvae. Reduction of populations of A. aegypti was high along the breeding season of this mosquito, being the effect less evident in C. pipiens. G. anceps could be an agent of control against container-breeding mosquitoes if its release in small water containers is complemented with other suitable management strategies.