Oxidative stress induction by essential oil from Piper alatipetiolatum (Piperaceae) triggers lethality in the larvae of Culex quinquefasciatus (Diptera: Culicidae).

Culex quinquefasciatus is the main vector of lymphatic filariasis in Brazil, which present resistance to commercial insecticides. Nowadays, essential oils (EOs) exhibiting larvicidal activity, such as those derived from Piper alatipetiolatum, provide a promising alternative for vector control, including Culex species. This study aimed to investigate the larvicidal activity and the oxidative stress indicators of the EO from P. alatipetiolatum in Cx. quinquefasciatus larvae. The EO was extracted from P. alatipetiolatum leaves using the hydrodistillation method, resulting in a yield of 7.2 ± 0.1%, analysed by gas chromatography coupled with spectrometry and gas chromatography coupled with flame ionization detector (GC-MS and GC-FID), and evaluated against Cx. quinquefasciatus larvae. Reactive Oxygen and Nitrogen Species (RONS), Catalase (CAT), glutathione-S-transferase (GST), acetylcholinesterase (AChE), and Thiol levels were used as oxidative stress indicators. Analysis by CG-MS and CG-FID revealed that the main compound in the EO was the oxygenated sesquiterpene ishwarone, constituting 78.6% of the composition. Furthermore, the EO exhibited larvicidal activity, ranging from 26 to 100%, with an LC50 of 4.53 µg/mL and LC90 of 15.37 µg/mL. This activity was accompanied by a significant increase in RONS production, alterations in CAT, GST, AChE activity, and thiol levels compared to the control groups (p < 0.05). To the best of our knowledge, this is the first report describing the larvicidal activity and oxidative stress induced by the EO from P. alatipetiolatum against Cx. quinquefasciatus larvae. Therefore, we propose that this EO shows promise as larvicidal agent for the effective control of Cx. quinquefasciatus larvae.

Identification of nectar sources foraged by female mosquitoes in Canada.

For many mosquito species, the females must obtain vertebrate blood to complete a gonotrophic cycle. These blood meals are frequently supplemented by feeding on sugary plant nectar, which sustains energy reserves needed for flight, mating, and overall fitness. Our understanding of mosquito nectar foraging behaviors is mostly limited to laboratory experiments and direct field observations, with little research into natural mosquito-host plant relationships done in North America. In this study, we collected nectar-fed female mosquitoes over a 2-year period in Manitoba, Canada, and amplified a fragment of the chloroplast rbcL gene to identify the plant species fed upon. We found that mosquitoes foraged from diverse plant families (e.g., grasses, trees, ornamentals, and legumes), but preferred certain species, most notably soybean and Kentucky blue grass. Moreover, there appeared to be some associations between plant feeding preferences and mosquito species, date of collection, landscape, and geographical region. Overall, this study implemented DNA barcoding to identify nectar sources forage by mosquitoes in the Canadian Prairies.

Development and laboratory validation of a plant-derived repellent blend, effective against Aedes aegypti [Diptera: Culicidae], Anopheles gambiae [Diptera: Culicidae] and Culex quinquefasciatus [Diptera: Culicidae].

Mosquitoes of the genera Aedes, Anopheles and Culex vector a wide range of pathogens seriously affecting humans and livestock on a global scale. Over-reliance on insecticides and repellents has driven research into alternative, naturally-derived compounds to fulfil the same objectives. Steam distilled extracts of four plants with strong, yet attractive, volatile profiles were initially assessed for repellency in a dual-port olfactometer using Aedes aegypti as the model species. Picea sitchensis was found to be the most repellent, proving comparable to leading products when applied at 100% (p = 1.000). Key components of conifer-derived volatile profiles were then screened via electroantennography before those components eliciting an electrophysiological response were assayed individually in the olfactometer; according to WHO protocol. The most promising 5 were selected for reductive analyses to produce an optimised semiochemical blend. This combination, and a further two variations of the blend, were then progressed to a multi-species analysis using the BG-test whereby bite-attempt frequency on hands was assessed under different repellent treatments; assays were compared between Aedes aegypti, Anopheles gambiae and Culex quinquefasciatus. Efficacy was found against all three species, although it was found that Ae. aegypti was the most susceptible to the repellent, with An. gambiae being the least. Here, a novel, naturally-derived blend is presented with weak spatial repellency, as confirmed in laboratory assays. Further work will be required to assess the full extent of the potential of the products, both in terms of field application and species screening; however, the success of the products developed demonstrate that plant metabolites have great capacity for use in the repellent sector; both to improve upon known compounds and to reduce the usage of toxic products currently on the market.

Sustainable use of plastic-derived nanocarbons as a promising larvicidal and growth inhibitor agent towards control of mosquitoes.

Nanoscale carbon was obtained from six widely used plastics (PET, HDPE, PVC, LDPE, PP and PP) via thermal degradation (600 °C) under inert atmosphere. The thermally degraded products were processed through bath sonication followed by lyophilisation and the same was characterized through proximate analysis, UV-Vis spectroscopy, Scanning electron micrograph (SEM) with energy dispersive X-ray (EDX) analysis, Transmission electron micrograph (TEM), Dynamic light scattering (DLS) and Fourier transform infrared spectroscopy (FTIR). A series of aqueous solution of nanoscale carbon (5-30 mg/L) were prepared and same were used as both mosquito growth inhibitor and larvicidal agent against 3rd and 4th instar larvae of Culex pipiens. The significant percent mortality results were recorded for LDPE (p < 0.007) with average particle size of 3.01 nm and 62.95 W% of carbon and PS (p < 0.002) with average particle size of 12.80 nm and 58.73 W% of carbon against 3rd instar larvae, respectively. Similarly, for 4th instar larvae, both significant pupicidal and adulticidal activity were also recorded for PET (F = 24.0, p < 0.0001 and F = 5.73, p < 0.006), and HDPE (F = 26.0, p < 0.0001) and F = 5.30, p < 0.008). However, significant pupicidal activity were observed for PVC (F = 6.90, p < 0.003), and PS (F = 21.30, p < 0.0001). Histological, bio-chemical and microscopic studies were revealed that nanoscale carbon causes mild to severe damage of external and internal cellular integrity of larvae. However, nanoscale carbon does not exhibit any chromosomal abnormality and anatomical irregularities in Allium cepa and Cicer arietinum, respectively. Similarly, non-significant results with respect to blood cell deformation were also recorded from blood smear of Poecilia reticulata. Therefore, it can be concluded that plastic origin nanoscale carbon could be a viable sustainable nano-weapon towards control of insects.

Relationships between water quality and mosquito presence and abundance: a systematic review and meta-analysis.

Mosquito-borne diseases (MBDs) are emerging in response to climate and land use changes. As mosquito (Diptera: Culicidae) habitat selection is often contingent on water availability for egg and larval development, studies have recognized water quality also influences larval habitats. However, underlying species-, genera-, and mosquito level preferences for water quality conditions are varied. This systematic review and meta-analysis aimed to identify, characterize, appraise, and synthesize available global data on the relationships between water quality and mosquito presence and abundance (MPA); with the goal to further our understanding of the geographic expansion of MBD risks. A systematic review was conducted to identify studies investigating the relationships between water quality properties and MPA. Where appropriate, random-effects meta-analyses were conducted to provide pooled estimates for the association between the most reported water quality properties and MPA. The most reported water quality parameters were pH (87%), nitrogen concentrations (56%), turbidity (56%), electrical conductivity (54%), dissolved oxygen (43%), phosphorus concentrations (30%), and alkalinity (10%). Overall, pH (P = 0.05), turbidity (P < 0.0001), electrical conductivity (P = 0.005), dissolved oxygen (P < 0.0001), nitrogen (P < 0.0001), and phosphorus (P < 0.0001) showed significantly positive pooled correlations with MPA, while alkalinity showed a nonsignificant null pooled correlation (P = 0.85). We observed high heterogeneity in most meta-analyses, and climate zonation was shown to influence the pooled estimates. Linkages between MPA and water quality properties will enhance our capacity to predict MBD risks under changing environmental and land use changes.

Vegetable oil-based surfactants are adjuvants that enhance the efficacy of neonicotinoid insecticides and can bias susceptibility testing in adult mosquitoes.

BACKGROUND: The standard operating procedure for testing the susceptibility of adult mosquitoes to neonicotinoid or butenolide insecticides recommends using a vegetable oil ester (Mero) as a surfactant. However, there is growing evidence that this adjuvant contains surfactants that can enhance insecticide activity, mask resistance and bias the bioassay. METHODOLOGY/PRINCIPAL FINDINGS: Using standard bioassays, we tested the effects of commercial formulations of vegetable oil-based surfactants similar to Mero on the activity of a spectrum of active ingredients including four neonicotinoids (acetamiprid, clothianidin, imidacloprid and thiamethoxam) and two pyrethroids (permethrin and deltamethrin). We found that three different brands of linseed oil soap used as cleaning products drastically enhanced neonicotinoid activity in Anopheles mosquitoes. At 1% (v/v), the surfactant reduced the median lethal concentration, LC50, of clothianidin more than 10-fold both in susceptible and in resistant populations of Anopheles gambiae. At 1% or 0.5% (v/v), linseed oil soap restored the susceptibility of adult mosquitoes fully to clothianidin, thiamethoxam and imidacloprid and partially to acetamiprid. By contrast, adding soap to the active ingredient did not significantly affect the level of resistance to permethrin or deltamethrin suggesting that vegetable oil-based surfactants specifically enhance the potency of some classes of insecticides. CONCLUSIONS/SIGNIFICANCE: Our findings indicate that surfactants are not inert ingredients, and their use in susceptibility testing may jeopardize the ability to detect resistance. Further research is needed to evaluate the potential, the limitations and the challenges of using some surfactants as adjuvants to enhance the potency of some chemicals applied in mosquito control.

Chemical Composition and Larvicidal Properties of Essential Oils from Wild and Cultivated Artemisia campestris L., an Endemic Plant in Morocco.

The Asteraceae family is well known for its toxic and repellent activity against mosquitoes. In this study, essential oils (EOs) extracted from the aerial parts of both wild and cultivated Artemisia campestris L. plants were tested for larvicidal activity against Culex pipiens (Diptera: Culicidae), a pest mosquito widely suspected to be the vector responsible for West Nile virus transmission. The research aims at comparing the chemical composition and insecticidal activity of cultivated and wild A. campestris EOs. The EOs were obtained by hydrodistillation from the plant's aerial parts and were analyzed using GC-MS. Furthermore, the larviciding experiment was carried out following the standard WHO protocol. The result showed that wild and cultivated plant EOs differed only quantitatively, while the qualitative profile revealed a nearly identical chemical composition. Camphor (18.98%), car-3-en-5-one (11.25%), thujone (6.36%), chrysanthenone (6.24%), filifolone (4.56%), and borneol (3.56%) dominate the wild plant EO. Camphor (21.01%), car-3-en-5-one (17%), chrysanthenone (10.15%), filifolone (7.90%), borneol (3.38%), and thujone (3.08%) are the major compounds of the cultivated plant. Cultivation did not affect the EO production since the yield of the cultivated plant was 0.5 ± 0.1% and 0.6 ± 0.2% for the wild plant. The cultivated A. campestris EO had the highest insecticidal activity (LC50 = 9.79 µg/ml), and no significant difference was noticed between wild and cultivated A. campestris EO in terms of LC90. These findings could pave the way for a new method of producing biocides to control major disease vectors and offer a potential alternative for pest control.

West Nile virus in India: An update on its genetic lineages.

West Nile virus (WNV) is a rapidly spreading mosquito-transmitted zoonotic flavivirus. Mosquitoes belonging to the genus Culex are incriminated as the principal vectors of the virus, which causes West Nile fever (WNF) in humans. Manifestations of WNF include a mild, self-limiting, flu-like illness, which in severe cases (rare) may progress to encephalitis, resulting in life-threatening consequences. WNV is geographically distributed worldwide, covering Africa, the Americas, Europe, and Asia (except Antarctica). The virus exists in a bird-mosquito transmission cycle in nature, with humans and horses as incidental/accidental hosts. The virus can infect a large variety of hosts worldwide, i.e., about 300 birds and around 70 different mosquito species belonging to several genera. For a long time, it was believed that WNV was not highly virulent and caused only mild infection globally. However, the recent frequent and increasing incidence of clinically severe WNV infections, such as encephalitis in humans and horses with significant mortality, has been reported in the Americas, Europe, and several East Asian countries. The emergence of lineage 2 strains endemic to Africa, with epidemic potential in humans and horses in Europe, is considered a serious global health concern. Although WNV is known to circulate in India since 1952, its re-emergence with severe neuro-invasive pathogenic potential in humans in Assam, Kerala, West Bengal and Tamil Nadu states signals urgent efforts to understand the dynamics of circulating strains with regard to its vector, hosts, and environment. This could be done by prioritizing "One Health" approach for developing effective preventive and control strategies. In view of the global interest, we present an overview of the circulating genetic lineages of WNV in India in comparison with the global scenario. In addition, we stress on holistic approaches of "One Health" strategy as the current need of the hour for designing effective preventive and control strategies in the country.

Maize pollen diet enhances malaria mosquito longevity and infectivity to Plasmodium parasites in Ethiopia.

Although larval diet quality may affect adult mosquito fitness, its impact on parasite development is scarce. Plant pollen from Zea mays, Typha latifolia, and Prosopis juliflora was ultraviolet-sterilized and examined for effects on larval development, pupation rate, adult mosquito longevity, survival and infectivity. The control larvae were fed Tetramin fish food as a comparator food. Four treatment and two control groups were used for each pollen diet, and each experimental tray had 25 larvae. Female An. arabiensis were starved overnight and exposed to infectious blood using a membrane-feeding system. The Kaplan-Meier curves and log-rank test were used for analysis. The Z. mays pollen diet increased malaria mosquito survival and pupation rate (91.3%) and adult emergence (85%). Zea mays and Tetramin fish food had comparable adulthood development times. Adults who emerged from larvae fed Z. mays pollen had the longest average wing length (3.72 mm) and were more permissive to P. vivax (45%) and P. falciparum (27.5%). They also survived longer after feeding on infectious blood and had the highest number of P. vivax oocysts. Zea mays pollen improved larval development, adult mosquito longevity, survival and infectivity to Plasmodium. Our findings suggest that malaria transmission in Z. mays growing villages should be monitored.

The Mosquito Larvicidal Activity of Lignans from Branches of Cinnamomum camphora chvar. Borneol.

The chemical investigation of branches of Cinnamomum camphora chvar. Borneol guided by mosquito larvicidal activity led to the isolation of fourteen known lignans (1-14). Their structures were elucidated unambiguously based on comprehensive spectroscopic analysis and comparison with the literature data. This is the first report of these compounds being isolated from branches of Cinnamomum camphora chvar. Borneol. Compounds 3-5 and 8-14 were isolated from this plant for the first time. All compounds isolated were subjected to anti-inflammatory, mosquito larvicidal activity and cytotoxic activity evaluation. Compounds (1-14) showed significant mosquito larvicidal activity against Culex pipiens quinquefasciatus with lethal mortality in 50% (LC50), with values ranging from 0.009 to 0.24 µg/mL. Among them, furofuran lignans(1-8) exhibited potent mosquito larvicidal activity against Cx. p. quinquefasciatus, with LC50 values of 0.009-0.021 µg/mL. From the perspective of a structure-activity relationship, compounds with a dioxolane group showed high mosquito larvicidal activity and have potential to be developed into a mosquitocide.

Development of a recombinant reporter Getah virus for antiviral drug screening assays.

Getah virus (GETV), is an often neglected and re-emerging mosquito-borne RNA virus. GETV can cause illness accompanied with high fever, rash, incapacitating arthralgia and chronic arthritis or encephalitic disease in affected animals. Currently, there is no specific treatment or vaccine against GETV infection. In this study, we developed three recombinant viruses by inserting different reporter protein genes between the Cap and pE2 genes. The reporter viruses exhibited high replication capacity similar to the parental virus. The rGECiLOV and rGECGFP viruses were genetically stable within at least ten rounds of passages in BHK-21 cells. We confirmed that the reporter virus, rGECGFP, facilitated the antiviral assays against GETV by testing it with the known inhibitor, ribavirin. It was also found that the compound, doxycycline, showed an inhibitory effect on GETV replication. In addition, rGECGFP was found to be an authentic mimic of the parental virus infection in 3-day-old mice, but with milder pathogenicity. The reporter viruses will contribute to the assessment of viral replication and proliferation, tracking and elucidating of alphavirus-host interactions. In addition, they will help in the screening of potential antiviral compounds.

Tracking antimalarial drug resistance using mosquito blood meals: a cross-sectional study.

BACKGROUND: Strong surveillance systems with wide geographic coverage are needed to detect and respond to reports of antimalarial drug resistance on the African continent. We aimed to assess the utility and feasibility of using blood-fed mosquitos (xenomonitoring) to conduct rapid surveillance of molecular markers associated with resistance in human populations. METHODS: We conducted three cross-sectional surveys in two rainy seasons and the interim dry season in southwest Burkina Faso between Oct 10, 2018, and Sept 17, 2019. We collected human blood samples and blood-fed mosquitos residing in household clusters across seven village sectors. Samples were assessed for Plasmodium falciparum with ultrasensitive quantitative PCR, genotyped for two markers of reduced drug susceptibility, pfmdr1 256A>T (Asn86Tyr) and pfcrt 227A>C (Lys76Thr), and sequenced for four markers of clonality. We assessed statistical equivalence using a 10% margin of equivalence. FINDINGS: We identified 551 infections in 1483 human blood samples (mean multiplicity of infection [MOI] 1·94, SD 1·47) and 346 infections in 2151 mosquito blood meals (mean MOI 2·2, SD 1·67). The frequency of pfmdr1 Asn86Tyr was 4% in survey 1, 2% in survey 2, and 12% in survey 3 in human samples, and 3% in survey 1, 0% in survey 2, and 8% in survey 3 in mosquito blood meals, and inter-host frequencies were statistically equivalent in surveys 1 and 2 (p<0·0001) but not Survey 3 (p=0·062) within a tolerability of 0·10. The frequency of pfcrt Lys76Thr was 16% in survey 1, 55% in survey 2, and 11% in survey 3 in humans and 40% in survey 1, 72% in survey 2, and 13% in survey 3 in mosquitos, and inter-host frequencies were equivalent in survey 3 only (p=0·032) within a tolerability of 0·10. In simulations, multiple but not preferential feeding behaviour in mosquitos reduced the accuracy of frequency estimates between hosts, particularly for markers circulating at higher frequencies. INTERPRETATION: Molecular markers in mosquito blood meals and in humans exhibited similar temporal trends but frequencies were not statistically equivalent in all scenarios. More work is needed to determine empirical and pragmatic thresholds of difference. Xenomonitoring might be an efficient tool to provide rapid information on emerging antimalarial resistance in regions with insufficient surveillance. FUNDING: National Institute of Allergy and Infectious Diseases. TRANSLATION: For the French translation of the abstract see Supplementary Materials section.

Bioactivity of brassica seed meals and its compounds as ecofriendly larvicides against mosquitoes.

Strategic, sustainable, and ecofriendly alternatives to chemical pesticides are needed to effectively control mosquitoes and reduce the incidence of their vectored diseases. We evaluated several Brassicaceae (mustard family) seed meals as sources of plant derived isothiocyanates produced from the enzymatic hydrolysis of biologically inactive glucosinolates for the control of Aedes aegypti (L., 1762). Five defatted seed meals (Brassica juncea (L) Czern., 1859, Lepidium sativum L., 1753, Sinapis alba L., 1753, Thlaspi arvense L., 1753, and Thlaspi arvense-heat inactivated and three major chemical products of enzymatic degradation (allyl isothiocyanate, benzyl isothiocyanate and 4-hydroxybenzyl isothiocyanate) were assayed to determine toxicity (LC50) to Ae. aegypti larvae. All seed meals except the heat inactivated T. arvense were toxic to mosquito larvae. L. sativum seed meal was the most toxic treatment to larvae (LC50 = 0.04 g/120 mL dH2O) at the 24-h exposure. At the 72-h evaluation, the LC50 values for B. juncea, S. alba and T. arvense seed meals were 0.05, 0.08 and 0.1 g/120 mL dH2O, respectively. Synthetic benzyl isothiocyanate was more toxic to larvae 24-h post treatment (LC50 = 5.29 ppm) compared with allyl isothiocyanate (LC50 = 19.35 ppm) and 4-hydroxybenzyl isothiocyanate (LC50 = 55.41 ppm). These results were consistent with the higher performance of the benzyl isothiocyanate producing L. sativum seed meal. Isothiocyanates produced from seed meals were more effective than the pure chemical compounds, based on calculated LC50 rates. Using seed meal may provide an effective method of delivery for mosquito control. This is the first report evaluating the efficacy of five Brassicaceae seed meals and their major chemical constituent against mosquito larvae and demonstrates how natural compounds from Brassicaceae seed meals can serve as a promising ecofriendly larvicides to control mosquitoes.

Chemistry and Bioactivity of Croton Essential Oils: Literature Survey and Croton hirtus from Vietnam.

Using essential oils to control vectors, intermediate hosts, and disease-causing microorganisms is a promising approach. The genus Croton in the family Euphorbiaceae is a large genus, with many species containing large amounts of essential oils, however, essential oil studies are limited in terms of the number of Croton species investigated. In this work, the aerial parts of C. hirtus growing wild in Vietnam were collected and analyzed by gas chromatography/mass spectrometry (GC/MS). A total of 141 compounds were identified in C. hirtus essential oil, in which sesquiterpenoids dominated, comprising 95.4%, including the main components ß-caryophyllene (32.8%), germacrene D (11.6%), ß-elemene (9.1%), α-humulene (8.5%), and caryophyllene oxide (5.0%). The essential oil of C. hirtus showed very strong biological activities against the larvae of four mosquito species with 24 h LC50 values in the range of 15.38-78.27 µg/mL, against Physella acuta adults with a 48 h LC50 value of 10.09 µg/mL, and against ATCC microorganisms with MIC values in the range of 8-16 µg/mL. In order to provide a comparison with previous works, a literature survey on the chemical composition, mosquito larvicidal, molluscicidal, antiparasitic, and antimicrobial activities of essential oils of Croton species was conducted. Seventy-two references (seventy articles and one book) out of a total of two hundred and forty-four references related to the chemical composition and bioactivity of essential oils of Croton species were used for this paper. The essential oils of some Croton species were characterized by their phenylpropanoid compounds. The experimental results of this research and the survey of the literature showed that Croton essential oils have the potential to be used to control mosquito-borne and mollusk-borne diseases, as well as microbial infections. Research on unstudied Croton species is needed to search for species with high essential oil contents and excellent biological activities.

Larvicidal evaluation of two novel cationic gemini surfactants against the potential vector of West Nile virus Culex pipiens Linnaeus (Diptera: Culicidae).

The development of insecticide resistance is a serious consequence of the widespread applications of synthetic insecticides. Recent studies have provided alternatives to currently available insecticides. Here, novel cationic gemini surfactants were synthesized to assess their insecticidal activities using laboratory and field strains larvae of Culex pipiens Linnaeus (Diptera: Culicidae). The efficacy of these surfactants was compared to that of clove oil and spinosad. The two surfactants G1 and G2 showed good insecticidal activities in laboratory strain with LC50 0.013 and 0.054 ppm, respectively, relative to spinosad with LC50 0.027 ppm, 48 h posttreatment. Although spinosad showed high efficiency against lab strain, it exhibited a high resistance ratio (RR) of 15.111 and 13.111 toward the field strain at 24 and 48 h posttreatment, respectively. The two gemini surfactants have a good safety profile and low RR (RR <5), which is close to clove oil; however, G1 and G2 presented high activities with 11,043.230 and 2658.648 folds, respectively, compared to clove oil. The treated Cx. pipiens larvae showed severe morphological malformations after treatment with gemini surfactants. The results of this study are promising in terms of developing novel, effective, affordable, and safe approaches for mosquito control strategies to reduce the risk of arbovirus transmission, which remains a global public health threat.

Larvicidal activity of hexane extract, fatty acids, and methyl esters from Tecoma stans pericarps against Culex quinquefasciatus.

Culex quinquefasciatus (Diptera: Culicidae) is responsible for the transmission of lymphatic filariasis. The search for natural sources of new insecticides to control mosquitoes has intensified in recent years. Tecoma stans is an exotic species in Brazil, known as 'ipê de jardim'. T. stans pericarps were extracted with hexane in Soxhlet apparatus, and fatty acids (FA) and methyl esters (FAME) were obtained by transesterification reaction of hexane extract (HE). HE, FA, and FAME were evaluated against 3rd. and 4th. instar larvae of Culex quinquefasciatus. Analysis by gas chromatography-mass spectrometry (GC/MS) identified the hydrocarbon nonacosane (81.69%) as the major compound in HE, and linolenic (16.89%), linoleic (16.83%), and palmitic acids (21.00%) were predominant in FA. FA and HE, at a concentration of 250 µg/mL, promoted mortality of 81.67% and 68.66% of larvae, respectively. HE and FA obtained from T. stans pericarps have larvicidal potential for the control of C. quinquefasciatus.

Larvicidal Effect of Hyptis suaveolens (L.) Poit. Essential Oil Nanoemulsion on Culex quinquefasciatus (Diptera: Culicidae).

Mosquitoes can be vectors of pathogens and transmit diseases to both animals and humans. Species of the genus Culex are part of the cycle of neglected diseases, especially Culex quinquefasciatus, which is an anthropophilic vector of lymphatic filariasis. Natural products can be an alternative to synthetic insecticides for vector control; however, the main issue is the poor water availability of some compounds from plant origin. In this context, nanoemulsions are kinetic stable delivery systems of great interest for lipophilic substances. The objective of this study was to investigate the larvicidal activity of the Hyptis suaveolens essential oil nanoemulsion on Cx. quinquefasciatus. The essential oil showed a predominance of monoterpenes with retention time (RT) lower than 15 min. The average size diameter of the emulsions (sorbitan monooleate/polysorbate 20) was ≤ 200 nm. The nanoemulsion showed high larvicidal activity in concentrations of 250 and 125 ppm. CL50 values were 102.41 (77.5253−149.14) ppm and 70.8105 (44.5282−109.811) ppm after 24 and 48 h, respectively. The mortality rate in the surfactant control was lower than 9%. Scanning micrograph images showed changes in the larvae's integument. This study achieved an active nanoemulsion on Cx. quinquefasciatus through a low-energy-input technique and without using potentially toxic organic solvents. Therefore, it expands the scope of possible applications of H. suaveolens essential oil in the production of high-added-value nanosystems for tropical disease vector control.

Plasmodium falciparum Sexual Commitment Rate Variation among Clinical Isolates and Diverse Laboratory-Adapted Lines.

Asexual blood-stage malaria parasites must produce sexual progeny to infect mosquitoes. It is important to understand the scope and causes of intraspecific variation in sexual commitment rates, particularly for the major human parasite P. falciparum. First, two alternative assay methods of measuring sexual commitment were compared to test a genetically modified P. falciparum line with elevated commitment rates inducible by overexpression of GDV1. The methods yielded correlated measurements with higher sensitivity and precision being achieved by one employing detection of the early gametocyte differentiation marker Pfs16. Thus, this was used to survey a diverse range of parasite lines and test each in multiple biological replicate assays in a serum-free medium supplemented with Albumax. There were differences among six recent clinical isolates from Ghana in their mean rates of sexual commitment per cycle, ranging from 3.3% to 12.2%. Among 13 diverse long-term laboratory-adapted lines, mean sexual commitment rates for most ranged from 4.7% to 13.4%, a few had lower rates with means from 0.3 to 1.6%, and one with a nonfunctional ap2-g gene always showed zero commitment. Among a subset of lines tested for the effects of exogenous choline to suppress commitment, there were significant differences. As expected, there was no effect in a line that had lost the gdv1 gene and that had generally low commitment, whereas the others showed quantitatively variable but significant responses to choline, suggesting potential trait variation. The results indicated the value of performing multiple replicate assays for understanding the variation of this key reproductive trait that likely affects transmission. IMPORTANCE Only sexual-stage malaria parasites are transmitted from human blood to mosquitoes. Thus, it is vital to understand variations in sexual commitment rates because these may be modifiable or susceptible to blocking. Two different methods of commitment rate measurement were first compared, demonstrating higher sensitivity and precision by the detection of an early differentiation marker, which was subsequently used to survey diverse lines. Clinical isolates from Ghana showed significant variation in mean per-cycle commitment rates and variation among biological replicates. Laboratory-adapted lines of diverse origins had a wider range with most being within the range observed for the clinical isolates, while a minority consistently had lower or zero rates. There was quantitative variation in the effects when adding choline to suppress commitment, indicating differing responsiveness of parasites to this environmental modification. Performing multiple assay replicates and comparisons of diverse isolates was important to understand this trait and its potential effects on transmission.

Mosquito Larvicides of Partial and Combinations Extract of Ethnobotanical Plant from North Sulawesi, Indonesia.

<b>Background and Objective:</b> Diseases caused by microbes vectored by mosquitoes are still a health problem in tropical countries today. DHF and Malaria are the two primary diseases vectored by mosquitoes, the morbidity and mortality rates have increased in low countries until now. However, the best way to control these two diseases is to control vectors, namely mosquitoes. Research has been conducted to determine the bioactive content and larvicidal activity of local plant extracts of North Sulawesi. <b>Materials and Methods:</b> The clove and trumpet flower samples were obtained from Minahasa, while the nutmeg samples were obtained from Sitaro Regency. Empirically, people use plant parts to repel mosquitoes. Extraction of plant simplicia was carried out by the maceration method. Qualitative and quantitative methods carried out the phytochemical content analysis. Qualitative analysis uses Harborne's (1996) method while qualitative analysis uses the UV Vis Spectrophotometer method. Toxicity tests were carried out on mosquito larvae developed in the laboratory. <b>Results:</b> The results showed that combining clove leaf extract, nutmeg flesh extract and trumpet flower synergistically increased the bioactive content. Flavonoids increased in the combination of extracts compared to partial extracts. The combination of extracts showed the highest toxicity to mosquito larvae (LC₅₀: 22.541 mg L¹), while the lowest was the partial extract of clove leaves with LC₅₀ (54.965 mg L¹). <b>Conclusion:</b> The combination of extracts showed the best toxicity activity on mosquito larvae. Research on bioactive characteristics and toxicity in adult mosquitoes needs to be carried out in the future.

Deciphering the role of polyphenol in defence mechanism against tea mosquito bug (Helopeltis theivora Waterhouse.) in cocoa (Theobroma cocoa L.).

Tea mosquito bug (TMB) is a serious pest of cocoa whose prevalence is high, mostly during summer and post monsoon season. Three species of tea mosquito bug have been reported on cocoa: Helopeltis antonii Signoret, H. theivora Waterhouse, and H. bradyi Waterhouse. H. theivora is the most prevalent one causing damage to young shoots, cherelles and pods. Rearing of tea mosquito bug on cocoa was found to be a failure in the present study hence Helopeltis theivora Waterhouse was maintained on the alternate host mile-a-minute (Mikania micrantha Kunth) under laboratory condition in insect rearing cages. Using freshly reared tea mosquito bugs twenty cocoa hybrids were screened for resistance and were ranked after 72 hours of screening. All the hybrids having less than three lesions per plant in seedlings and less than 33 lesions on pods were ranked as highly resistant. It was observed that hybrids classified as highly resistant had significantly higher phenol content than those classified as susceptible. The significantly low phenol content in the susceptible hybrids suggests that phenolics have a function in mediating resistance to tea mosquito bug in cocoa. From correlation and regression analysis it is confirmed that phenol content can be used as a potential marker indicating the level of resistance of cocoa hybrids against tea mosquito bug resistance.