Adult Anopheles Research Articles

Repellency Effects, Cytotoxicity Assessment and Chemical Compositions of Citrus limon Seeds Extracts and Fractions Against Anopheles gambiae sl Mosquitoes

Background: Citrus limon seeds have limited exploration, compared to other parts. The increasing prevalence of mosquito menace prompted the investigation into the repellent effects of the seed n-hexane extract and fractions of Citrus limon (Rutaceae) against adult Anopheles gambiae sl mosquitoes. The cytotoxicity and chemical constituents of the 100% n-Hexane Fraction (HF) were also evaluated. Methods: Successive extraction was done with cold-maceration using, n-hexane, Ethyl Acetate (EA) and methanol. The three solvent extracts were screened against Anopheles gambiae sl mosquitos, using human bait method, at doses, 1.5, 2.5 and 5.0 mg/ml and with Deet and acetone used as positive and negative controls respectively. The n-hexane extract was further fractionated with column chromatography. Ten fractions (100% n-hexane, 10, 20, 30, 40, 60, 80% EA in n-hexane, 100% EA, 50% methanol in EA, and 100% methanol) were collected, and all tested at a single optimized dose of 5 mg/ml for 2 hr. The cytotoxicity and the chemical constituents of HF were determined using brine shrimp lethality test and gas chromatography-mass spectrometry respectively. Results: N-hexane extract and HF, both at 5 mg/ml, showed the most protective effect of 97.52 and 89.40% respectively, compared with standard (Odomos Deet) 100% activity, with no significant difference (p>0.05) between the three. HF was toxic with LC50 values of 29.2354 ppm. Thirty compounds representing 79.25% of HF were identified. Major compounds present in the fraction were heptadecanamine (8.31%), N-(3-methylbutyl) acetamide (7.89%), 2-ethyl- 1-hexanamine (6.65%). Conclusion: This promising repellent activity further justified the traditional usage of Citrus limon as mosquito repellent.

Insecticide resistant Anopheles from Ethiopia but not Burkina Faso show a microbiota composition shift upon insecticide exposure

BackgroundMalaria remains a key contributor to mortality and morbidity across Africa, with the highest burden in children under 5. Insecticide-based vector control tools, which target the adult Anopheles mosquitoes, are the most efficacious tool in disease prevention. Due to the widespread use of these interventions, insecticide resistance to the most used classes of insecticides is now pervasive across Africa. Understanding the underlying mechanisms contributing to this phenotype is necessary to both track the spread of resistance and to design new tools to overcome it.MethodsHere, we compare the microbiota composition of insecticide-resistant populations of Anopheles gambiae, An. coluzzii and An. arabiensis from Burkina Faso, and in the latter case additionally from Ethiopia, to insecticide-susceptible populations.ResultsWe show that the microbiota composition between insecticide-resistant and -susceptible populations does not differ in Burkina Faso. This result is supported by data from laboratory colonies originating in Burkina Faso across two countries. In contrast, An. arabiensis from Ethiopia demonstrates clear differences in microbiota composition in those dying from and those surviving insecticide exposure. To further understand resistance in this An. arabiensis population, we performed RNAseq and saw differential expression of detoxification genes associated with insecticide resistance and changes in respiration, metabolism and synapse-related ion channels.ConclusionsOur results indicate that, in addition to changes in the transcriptome, microbiota can contribute to insecticide resistance in certain settings.Graphical

Risk of Aedes-borne diseases in and around the Tanzanian seaport of Tanga despite community members being more concerned about malaria

BackgroundIncreased global trade, while beneficial economically, can also increase the spread of vector-borne diseases, particularly those transmitted by Aedes mosquitoes spreading via trade routes. Given the heightened trade-induced activity at ports of entry, it is particularly crucial to assess the risk of mosquito-borne diseases in these settings. This study compared the risks of Aedes-borne disease in and around the eastern Tanzanian seaport of Tanga.MethodsA 200 m × 200 m grid-based system was used to sample mosquitoes within the port area, and in surrounding areas at 2 km, 2.5 km, and 5 km away, between June and December 2023. We characterized mosquito breeding habitats, collected mosquito larvae using standard dippers and tested susceptibility of raised adult Aedes aegypti populations to different insecticides. Adult mosquitoes were collected using BG sentinel traps (daytime) and Centers for Disease Control (CDC) light traps (night-time). Additionally, more than 200 port users and neighboring residents were surveyed to assess their experiences with and perceptions of mosquito biting and disease risks.ResultsThere were 2931 breeding sites, with (60.8%, n = 1782) positive for Aedes larvae. The percentage of water-holding containers infested with Aedes immatures, i.e., the container index (CI), was highest in the port area (66.2%), and lowest 5 km away (44.6%). The port area also had a greater proportion of temporary breeding sites (64.9%) than did the surrounding areas. The adult mosquito surveys revealed 20,449 mosquito species including: Culex quinquefasciatus (56.2%), Mansonia uniformis (38.6%), Ae. aegypti (5.1%), Anopheles gambiae (0.1%), and Anopheles funestus. Ae. aegypti were more abundant in the port area than in the surrounding areas (P < 0.001), whereas Culex sp., and Mansonia sp., were significantly outside (P < 0.001). Adult Anopheles sp., were found only in the port area, but Anopheles larvae were found both within and outside the port areas. Tests on Ae. aegypti sp., revealed susceptibility to bendiocarb and DDT, and resistance to permethrin. Awareness of mosquito-borne diseases among respondents was high for malaria (64.8%), but low for dengue (26.3%) and Chikungunya (1.7%). Most respondents reported being bothered by mosquitoes mostly at night (53.4%) or in the evening (40.7%). In addition to insecticidal bednets, which are used primarily against malaria, preventive measures for Aedes-borne diseases are limited.ConclusionsThis study identified significant potential risk of Aedes species, specifically Ae. aegypti sp., and associated diseases, but low perception of risk and inadequate personal protection measures in the study area. This low perception of risk highlights the need to improve public knowledge of the transmission and control of Aedes-borne diseases.Graphical

Mosquito breeding water parameters are important determinants for Microsporidia MB in the aquatic stages of Anopheles species

BackgroundMicrosporidia MB disrupts Plasmodium development in Anopheles mosquitoes, making it a possible biocontrol tool for malaria. As a tool for vector/disease control, its ecological distribution and the factors that determine their occurrence must be defined. We investigated the frequency of Microsporidia MB in Anopheles mosquitoes across selected sites in northern and southern Ghana, as well as the physicochemical parameters of mosquito breeding water that are associated with the occurrence of the fungus, by fitting regression models.MethodsA non-column extraction method was used to extract DNA from the abdomens of 4255 adult Anopheles mosquitoes that emerged from larvae and pupae collected between August and October of 2021 and 2022. Detection of Microsporidia MB was achieved using quantitative PCR (qPCR), while mosquito species were molecularly identified using short interspersed nuclear elements (SINE), restriction fragment length polymorphism (RFLP) methods, and the ANOSPP algorithm.ResultsOverall Microsporidia MB distribution was 2.2% (92/4255). Male mosquitoes exhibited a higher frequency of infections and had a predicted probability of infection that was 85% higher than that of females. Sites in Ghana's Savannah zone had the highest Microsporidia MB distribution (68.5%). Biochemical oxygen demand in mosquito breeding water was estimated to be positively associated with and significantly predicts Microsporidia MB in mosquitoes with an accuracy of 94%. Increasing ammonium ion concentrations reduced the chances of finding Microsporidia MB-positive mosquitoes. According to our data, all Anopheles mosquitoes, including minor species such as An. squamosus, An. pretoriensis and An. rufipes, had equal probability of Microsporidia MB infection.ConclusionsThese results provide preliminary information on micro-ecological factors that potentially support the sustainability of Microsporidia MB infection in mosquitoes during their aquatic life stages. It will be important, therefore, to explore the impact of strategies for larval source management on these factors to ensure that the symbiont's persistence during the host's aquatic stages may not be adversely affected should it be used as an integrated approach for mosquito/disease control.Graphical abstract

A survey of malaria vectors feeding preference, biting site and resting behaviour in the malaria elimination settings of Dembiya District, north-western Ethiopia.

Despite the progress in scaling vector control interventions in Ethiopia, malaria is still a major health problem in the country. Monitoring of the local vector populations and the effectiveness of vector control strategies is necessary to guide programme decisions to optimize malaria prevention efforts. This study investigated the feeding preference, the biting behaviour and resting behaviours of Anopheles mosquitoes in selected localities of Dembiya District. Adult Anopheles mosquitoes were sampled indoors and outdoors from June 2018 to May 2019 using CDC light traps, pyrethrum spray catches, artificial pit shelters, and mouth aspirators at both Guramba Bata and Arebiya study sites. Anopheles mosquitoes were identified to the species level. Their blood meal source and Plasmodium sporozoite infections were determined using an enzyme-linked immunosorbent assay. Anopheles mosquitoes belonging to 11 species were identified from 2,055 collected mosquito specimens. Anopheles pharoensis was the predominant species at both the Guramba Bata (46.5%) and Arebiya (46.2%) study sites. The CDC light traps caught the highest number of Anopheles mosquitoes in both study sites. In Guramba Bata the density of outdoor host-seeking and resting Anopheles mosquitoes were higher than indoors (P ≤ 0.05). The human blood indexes (HBI) of indoor and outdoor host-seeking Anopheles arabiensis were 17.4% and 15.3%, respectively. The entomological inoculation rate (EIR) of outdoor host-seeking An. arabiensis was 4.7 infective bites/person/year. Additionally, the outdoor EIR of host-seeking Anopheles coustani was 25.7ib/p/year. Anopheles mosquitoes in Dembiya district were more likely to seek a host and rest outdoors than indoors. A reevaluation of vector control strategies is needed to ensure Ethiopia remains on the path to malaria elimination. The detection of Plasmodium circumsporozoite protein in potential secondary vectors, such as An. coustani requires further investigation to substantiate their role in malaria transmission.

Contrasting patterns of Asaia association with Plasmodium falciparum between field-collected Anopheles gambiae and Anopheles coluzzii from Cameroon.

The widespread prevalence of Asaia in mosquitoes makes it a potential candidate for paratrangenic control in Anopheles. To better understand whether this bacterium could be used for malaria control, we quantified Asaia in An. gambiae s.l populations in malaria endemic regions examining co-infection with Plasmodium falciparum. Adult Anopheles mosquitoes were collected across two different eco-geographical localities in Cameroon, during both the dry and wet seasons. DNA was extracted from whole individual mosquitoes, and real time-qPCR amplification of the 16S ribosomal RNA was used to quantify Asaia in both An. gambiae and An. coluzzii samples. We also detected and quantified P. falciparum infection in the same mosquitoes. The density of Asaia was successfully quantified in a total of 864 field mosquitoes, comprising of 439 An. gambiae from Bankeng and 424 An. coluzii collected from Gounougou. Interestingly, a higher prevalence of Asaia in An. gambiae (88.3%) compared to An. coluzzii (80.9%) was observed. Moreover, the density of Asaia in both species was significantly affected by seasonal changes in the two localities. Furthermore, a significant difference between the infection densities of Asaia and the Plasmodium infection status in the two species was recorded. However, no correlation was observed between the number of Asaia and P. falciparum infections. This study provides evidence that naturally occurring Asaia infection is not correlated to P. falciparum development within An. gambiae and An. coluzzii. Nevertheless, further studies incorporating experimental infections are required to better investigate the correlation between Anopheles mosquitoes, Asaia, and Plasmodium.IMPORTANCEThe symbiont Asaia has emerged as a promising candidate for paratransgenic control of malaria, but further analysis of its biology and genetics across Africa is necessary. In this study, we investigated and quantified the influence of Asaia in naturally infected An. gambiae s.l. populations with the malaria parasite Plasmodium falciparum. Genomic DNA was extracted from whole individual mosquitoes collected from two localities, and Asaia was quantified using real-time qPCR by amplification of the 16S ribosomal RNA gene. We also detected and quantified Plasmodium falciparum infection in the same mosquitoes and established the correlation between Asaia and Plasmodium coinfection. This study provides evidence that naturally occurring Asaia infection is not correlated with P. falciparum development within An. gambiae and An. coluzzii mosquitoes.

Pathogenicity and sub-lethal activity of orally administered entomopathogenic fungi against two adult mosquito species, Aedes aegypti (Diptera: Culicidae) and Anopheles stephensi (Diptera: Culicidae)

Entomopathogenic fungi (EPF) are known for their efficacy in controlling adult mosquito populations by penetrating through their cuticle. However, the effect of oral administration of EPF on the biological parameters of Aedes aegypti and Anopheles stephensi remains largely unexplored. This study aimed to assess the effect of orally administrated EPF isolates on the survival, feeding behavior, fecundity, fertility, follicle development and host-searching behavior in response to yeast-generated CO2 of Ae. aegypti and An. stephensi. An initial screening of 50 isolates involved exposure of adult Ae. aegypti and An. stephensi by integument inoculation. Subsequently, the entomopathogenic effect of the five highly virulent isolates was confirmed through oral administration revealing Beauveria pseudobassiana 42–51 as a potent mosquito killer. B. pseudobassiana 42–51 was administered orally to evaluate sub-lethal effects. The results showed a 63 % and 43 % reduction in blood feeding of Ae. aegypti and An. stephensi, respectively. Furthermore, a decrease in egg hatching rate was observed, with a reduction of 83% for Ae. aegypti and 74% for An. stephensi on the seventh day following fungal administration, showing decreased hatchability in both species. Poor and abnormal follicle development was observed in both mosquito species. Also, the host-searching behavior was evaluated by attraction to CO2 utilizing a Y-tube olfactometer. A tendency of reduction in the attraction rate towards the odor was observed three days post-fungal administration. These findings underscore the significant impact of oral administration of B. pseudobassiana 42–51 on mosquitoes, highlighting not only its lethal effects but also sub lethal impacts on their biology. Moreover, this fungus may exhibit the potential to simultaneously control both mosquito species and serve as a biocontrol agent for the management of vector-borne diseases.

A study on the effectiveness of (+)-usnic acid as oral toxic sugar bait against adult male and female Anopheles gambiae

BackgroundDespite the application of various tools for the control of vectors of Plasmodium falciparum, malaria remains the major killer disease in sub-Saharan Africa accounting for up to 90% of deaths due to the disease. Due to limitations of the useage of chemical insecticides such as resistance, negative impact on the environment and to nontarget organisms, the World Health Organization (WHO) requires that affected countries find alternative vector control tools. This study evaluated the effectiveness of ( +)-usnic acid (UA) as an insecticide through oral administration to male and female Anopheles gambiae as an alternative or additional active ingredient to be used in toxic sugar bait.Methods( +)-usnic acid was diluted using acetone at 5, 10, and 15 mg/ml concentrations in three replicates. A 5 ml mixture of 2% food dye and 10% sugar using chlorine-free water mixed with the dilutions of the ( +)-usnic acid and negative control was made containing 2% food dye and 10% sugar solution. The preparations were soaked on a ball of cotton wool and placed over the net of a cup. 5 male and 5 non-blood-fed female newly hatched starved An. gambiae Kisumu strain were introduced together into a cup and monitored for knockdown and mortalities after 4, 24 48, and 72 h. The data were analysed using a multiple linear regression model using the lm function, a base R function and a posthoc test were conducted on the significant main effects and interaction terms using the emmeans function from the emmeans R package. All analyses were performed in RStudio using base R (version 4.3.3).ResultsThere was high mortality of both male and female An. gambiae after ingestion of the toxic sugar bait. 15 mg/ml usnic acid caused the highest mortality (50%) within the first 4 h compared to 5 and 10 mg/ml ( +)-UA. There was a decline in the mortality rate with increased exposure time from 24 to 72 h, however, there was a significant difference in mortality at 5, 10 and 15 mg/ml. Acute toxicity was associated with ingestion of 15 mg/ml after 24 h. 72 h post-mortality was lower in all concentrations than in the control. High mortality was observed among females over the first 4 h (60%) compared to males (40%) due to higher feeding rate of the toxic agent. The proportion of dead males and females was equal after 24 h while after 48 h, the proportion of dead males was high.There was a significantly lower mortality rate after 72 h for both males and females (0 to 13.3%). Compared to all the treatments, high mortality of males was observed.ConclusionsThe results of this study indicate that ( +)-UA when administered as oral sugar bait to An. gambiae has insecticidal properties and is a suitable ingredient to be used as a toxic agent in the novel attractive toxic sugar bait for the control of malaria vectors. ( +)-UA may be an alternative active ingredient as toxic bait in the effort to reduce and eliminate the transmission of Plasmodium falciparum in Africa.

Urban malaria and its determinants in Eastern Ethiopia: the role of Anopheles stephensi and urbanization

BackgroundMalaria prevention and control strategies have been hampered by urbanization and the spread of Anopheles stephensi. The spread of this vector into Africa further complicates the already complex malaria situation, that could put about 126 million Africans at risk of infection. Hence, this study aimed to assess the determinants of urban malaria, focusing on the role of urbanization and the distribution of An. stephensi in Eastern Ethiopia.MethodsA matched case control study was conducted among febrile urban residents of Dire Dawa (malaria positive as cases and negative as a control). A capillary blood sample was collected for parasite identification using microscopic examination and an interviewer administered questionnaire was used to collect additional data. Centers for Disease Control and Prevention miniature light traps (CDC-LT) and Prokopack aspirator were used to collect adult mosquito vectors from the selected cases and control houses to identify the mosquito vector species. Then, the data were exported to STATA for analysis. Conditional logistic regression was done to identify determinants, and principal component Analysis (PCA) was done for some independent variables.ResultsThis study enrolled 132 cases and 264 controls from urban setting only. Of the 132 cases, 90 cases were positive for Plasmodium falciparum, 34 were positive for Plasmodium vivax and 8 had mixed infections. All cases and controls were similar with regard to their respective age and sex. Travel history (AOR: 13.1, 95% CI 2.8–61.4), presence of eves and holes on walls (AOR: 2.84, 95% CI 1.5–5.5), history of malaria diagnosis (AOR: 2.4, 95% CI 1.1–5.3), owning any livestock (AOR: 7.5, 95% CI 2.4–22.8), presence of stagnant water in the area (AOR: 3.2, 95% CI 1.7–6.1), sleeping under bed net the previous night (AOR: 0.21, 95% CI 0.1–0.6) and knowledge on malaria and its prevention (AOR: 2.2, 95% CI 1.2–4.1) were determinants of urban malaria infection. About 34 adult Anopheles mosquitoes were collected and identified from those selected cases and control houses and 27 of them were identified as An. stephensi.ConclusionAmong the cases, the dominant species were P. falciparum. This study identified travel history, house condition, past infection, livestock ownership, stagnant water, bed net use, and malaria knowledge as determinants of infection. This study also found the dominance of the presence of An. stephensi among the collected mosquito vectors. This suggests that the spread of An. stephensi may be impacting malaria infection in the study area. Hence, strengthening urban-targeted malaria interventions should be enhanced to prevent and control further urban malaria infection and spread.

The impact of Bacillus thuringiensis var. israelensis (Vectobac® WDG) larvicide sprayed with drones on the bio-control of malaria vectors in rice fields of sub-urban Kigali, Rwanda

BackgroundThe core vector control tools used to reduce malaria prevalence are currently long-lasting insecticidal nets (LLINs), and indoor residual spraying (IRS). These interventions are hindered by insecticide resistance and behavioural adaptation by malaria vectors. Thus, for effective interruption of malaria transmission, there is a need to develop novel vector control interventions and technologies to address the above challenges. Larviciding using drones was experimented as an innovative tool that could complement existing indoor interventions to control malaria.MethodsA non-randomized larviciding trial was carried out in irrigated rice fields in sub-urban Kigali, Rwanda. Potential mosquito larval habitats in study sites were mapped and subsequently sprayed using multirotor drones. Application of Bacillus thuringiensis var. israelensis (Bti) (Vectobac® WDG) was followed by entomological surveys that were performed every two weeks over a ten-month period. Sampling of mosquito larvae was done with dippers while adult mosquitoes were collected using CDC miniature light traps (CDC-LT) and pyrethrum spraying collection (PSC) methods. Malaria cases were routinely monitored through community health workers in villages surrounding the study sites.ResultsThe abundance of all-species mosquito larvae, Anopheles larvae and all-species pupae declined by 68.1%, 74.6% and 99.6%, respectively. Larval density was reduced by 93.3% for total larvae, 95.3% for the Anopheles larvae and 61.9% for pupae. The total adult mosquitoes and Anopheles gambiae sensu lato collected using CDC-Light trap declined by 60.6% and 80% respectively. Malaria incidence also declined significantly between intervention and control sites (U = 20, z = − 2.268, p = 0.023).ConclusionsThe larviciding using drone technology implemented in Rwanda demonstrated a substantial reduction in abundance and density of mosquito larvae and, concomitant decline in adult mosquito populations and malaria incidences in villages contingent to the treatment sites. The scaling up of larval source management (LSM) has to be integrated in malaria programmes in targeted areas of malaria transmission in order to enhance the gains in malaria control.

Plasmodium gametocyte carriage in humans and sporozoite rate in anopheline mosquitoes in Gondar zuria district, Northwest Ethiopia.

Although the overall burden of malaria is decreasing in Ethiopia, a recent report of an unpredictable increased incidence may be related to the presence of community-wide gametocyte-carrier individuals and a high proportion of infected vectors. This study aimed to reveal the current prevalence of gametocyte-carriage and the sporozoite infectivity rate of Anopheles vectors for Plasmodium parasites. A community-based cross-sectional study was conducted from May 01 to June 30/2019. A total of 53 households were selected using systematic random sampling and a 242 study participants were recruited. Additionally,515 adult female Anopheles mosquitoes were collected using Center for Diseases Control and Prevention (CDC) light traps and mouth aspirators. Parasite gametocytemia was determined using giemsa stain microscopy, while sporozoite infection was determined by giemsa staining microscopy and enzyme linked immunosorbent assay (ELISA). Among the total 242 study participants, 5.4% (95%, CI = 2.9-8.3) of them were positive for any of the Plasmodium species gametocyte. Furthermore, being female [AOR = 15.5(95%, CI = 1.71-140.39)], age group between 15-29 years old [AOR = 16.914 (95%, CI = 1.781-160.63)], no ITNs utilization [AOR = 16.7(95%, CI = 1.902 -146.727)], and high asexual parasite density [(95%, CI = 0.057-0.176, P = 0.001, F = 18.402)] were identified as statistically significant factors for gametocyte carriage. Whereas sporozoite infection rate was 11.6% (95%, CI = 8.2-15.5) and 12.7% (95%, CI = 9.6-16.3) by microscopy and ELISA, respectively. Overall, this study indicated that malaria remains to be an important public health problem in Gondar Zuria district where high gametocyte carriage rate and sporozoite infection rate could sustain its transmission and burden. Therefore, in Ethiopia, where malaria elimination program is underway, frequent, and active community-based surveillance of gametocytemia and sporozoite infection rate is important.

Residual bioefficacy of attractive targeted sugar bait stations targeting malaria vectors during seasonal deployment in Western Province of Zambia

BackgroundThe primary vector control interventions in Zambia are long-lasting insecticidal nets and indoor residual spraying. Challenges with these interventions include insecticide resistance and the outdoor biting and resting behaviours of many Anopheles mosquitoes. Therefore, new vector control tools targeting additional mosquito behaviours are needed to interrupt transmission. Attractive targeted sugar bait (ATSB) stations, which exploit the sugar feeding behaviours of mosquitoes, may help in this role. This study evaluated the residual laboratory bioefficacy of Westham prototype ATSB® Sarabi v.1.2.1 Bait Station (Westham Ltd., Hod-Hasharon, Israel) in killing malaria vectors in Western Province, Zambia, during the first year of a large cluster randomized phase-III trial (Clinical Trials.gov Identifier: NCT04800055).MethodsThis was a repeat cross-sectional study conducted within three districts, Nkeyema, Kaoma, and Luampa, in Western Province, Zambia. The study was conducted in 12 intervention clusters among the 70 trial clusters (35 interventions, 35 controls) between December 2021 and June 2022. Twelve undamaged bait stations installed on the outer walls of households were collected monthly (one per cluster per month) for bioassays utilizing adult female and male Anopheles gambiae sensu stricto (Kisumu strain) mosquitoes from a laboratory colony.ResultsA total of 84 field-deployed ATSB stations were collected, and 71 ultimately met the study inclusion criteria for remaining in good condition. Field-deployed stations that remained in good condition (intact, non-depleted of bait, and free of dirt as well as mold) retained high levels of bioefficacy (mean induced mortality of 95.3% in males, 71.3% in females, 83.9% combined total) over seven months in the field but did induce lower mortality rates than non-deployed ATSB stations (mean induced mortality of 96.4% in males, 87.0% in females, 91.4% combined total). There was relatively little variation in corrected mortality rates between monthly rounds for those ATSB stations that had been deployed to the field.ConclusionWhile field-deployed ATSB stations induced lower mortality rates than non-deployed ATSB stations, these stations nonetheless retained relatively high and stable levels of bioefficacy across the 7-month malaria transmission season. While overall mean mosquito mortality rates exceeded 80%, mean mortality rates for females were 24 percentage points lower than among males and these differences merit attention and further evaluation in future studies. The duration of deployment was not associated with lower bioefficacy. Westham prototype ATSB stations can still retain bioefficacy even after deployment in the field for 7 months, provided they do not meet predetermined criteria for replacement.

An ecotoxicological view on malaria vector control with ivermectin-treated cattle

Malaria remains an enduring challenge in sub-Saharan Africa, affecting public health and development. Control measures can include the use of insecticides that target adult Anopheles mosquitoes transmitting the malaria-causing Plasmodium parasite. Such mosquitoes can also bite livestock, allowing vector populations to be maintained at levels that enable parasite transmission. Thus, one way to control the spread of malaria includes the use of endectocide-treated livestock which renders the blood of cattle toxic to such mosquito populations. Here we present an ecotoxicological perspective on malaria vector control, using cattle treated with the endectocide ivermectin to target zoophagic and opportunistic Anopheles coluzzii mosquitoes. Our study employs an innovative, long-acting injectable ivermectin formulation with over 6 months of sustained mosquitocidal activity. Robust vector population modelling underscores its promising field effectiveness. Environmental implications (soil sorption and dissipation) of excreted ivermectin and potential ecotoxicological risks to non-target dung organisms in West Africa are discussed, in addition to actionable, locally inspired risk mitigation measures to protect sub-Saharan soils and agroecosystems from chemical pollution. We highlight how ecotoxicology and environmental chemistry improve livestock-based vector control with ivermectin for effective and more sustainable malaria management.

Measuring effects of ivermectin-treated cattle on potential malaria vectors in Vietnam: A cluster-randomized trial.

Malaria elimination using current tools has stalled in many areas. Ivermectin (IVM) is a broad-antiparasitic drug and mosquitocide and has been proposed as a tool for accelerating progress towards malaria elimination. Under laboratory conditions, IVM has been shown to reduce the survival of adult Anopheles populations that have fed on IVM-treated mammals. Treating cattle with IVM has been proposed as an important contribution to malaria vector management, however, the impacts of IVM in this One Health use case have been untested in field trials in Southeast Asia. Through a randomized village-based trial, this study quantified the effect of IVM-treated cattle on anopheline populations in treated vs. untreated villages in Central Vietnam. Local zebu cattle in six rural villages were included in this study. In three villages, cattle were treated with IVM at established veterinary dosages, and in three additional villages cattle were left as untreated controls. For the main study outcome, the mosquito populations in all villages were sampled using cattle-baited traps for six nights before, and six nights after a 2-day IVM-administration (intervention) period. Anopheline species were characterized using taxonomic keys. The impact of the intervention was analyzed using a difference-in-differences (DID) approach with generalized estimating equations (with negative binomial distribution and robust errors). This intervention was powered to detect a 50% reduction in total nightly Anopheles spp. vector catches from cattle-baited traps. Given the unusual diversity in anopheline populations, exploratory analyses examined taxon-level differences in the ecological population diversity. Across the treated villages, 1,112 of 1,523 censused cows (73% overall; range 67% to 83%) were treated with IVM. In both control and treated villages, there was a 30% to 40% decrease in total anophelines captured in the post-intervention period as compared to the pre-intervention period. In the control villages, there were 1,873 captured pre-intervention and 1,079 captured during the post-intervention period. In the treated villages, there were 1,594 captured pre-intervention, and 1,101 captured during the post-intervention period. The difference in differences model analysis comparing total captures between arms was not statistically significant (p = 0.61). Secondary outcomes of vector population diversity found that in three villages (one control and two treatment) Brillouin's index increased, and in three villages (two control and one treatment) Brillouin's index decreased. When examining biodiversity by trapping-night, there were no clear trends in treated or untreated vector populations. Additionally, there were no clear trends when examining the components of biodiversity: richness and evenness. The ability of this study to quantify the impacts of IVM treatment was limited due to unexpectedly large spatiotemporal variability in trapping rates; an area-wide decrease in trapping counts across all six villages post-intervention; and potential spillover effects. However, this study provides important data to directly inform future studies in the GMS and beyond for IVM-based vector control.

Mapping malaria vectors and insecticide resistance in a high-endemic district of Haryana, India: implications for vector control strategies

BackgroundAchieving effective control and elimination of malaria in endemic regions necessitates a comprehensive understanding of local mosquito species responsible for malaria transmission and their susceptibility to insecticides.MethodsThe study was conducted in the highly malaria prone Ujina Primary Health Center of Nuh (Mewat) district of Haryana state of India. Monthly entomological surveys were carried out for adult mosquito collections via indoor resting collections, light trap collections, and pyrethrum spray collections. Larvae were also collected from different breeding sites prevalent in the region. Insecticide resistance bioassay, vector incrimination, blood meal analysis was done with the collected vector mosquitoes.ResultsA total of 34,974 adult Anopheles mosquitoes were caught during the survey period, out of which Anopheles subpictus was predominant (54.7%). Among vectors, Anopheles stephensi was predominant (15.5%) followed by Anopheles culicifacies (10.1%). The Human Blood Index (HBI) in the case of An. culicifacies and An. stephensi was 6.66 and 9.09, respectively. Vector incrimination results revealed Plasmodium vivax positivity rate of 1.6% for An. culicifacies. Both the vector species were found resistant to DDT, malathion and deltamethrin.ConclusionThe emergence of insecticide resistance in both vector species, compromises the effectiveness of commonly used public health insecticides. Consequently, the implementation of robust insecticide resistance management strategies becomes imperative. To effectively tackle the malaria transmission, a significant shift in vector control strategies is warranted, with careful consideration and adaptation to address specific challenges encountered in malaria elimination efforts.

Spatiotemporal distribution and bionomics of Anopheles stephensi in different eco-epidemiological settings in Ethiopia

BackgroundMalaria is a major public health concern in Ethiopia, and its incidence could worsen with the spread of the invasive mosquito species Anopheles stephensi in the country. This study aimed to provide updates on the distribution of An. stephensi and likely household exposure in Ethiopia.MethodsEntomological surveillance was performed in 26 urban settings in Ethiopia from 2021 to 2023. A kilometer-by-kilometer quadrant was established per town, and approximately 20 structures per quadrant were surveyed every 3 months. Additional extensive sampling was conducted in 50 randomly selected structures in four urban centers in 2022 and 2023 to assess households’ exposure to An. stephensi. Prokopack aspirators and CDC light traps were used to collect adult mosquitoes, and standard dippers were used to collect immature stages. The collected mosquitoes were identified to species level by morphological keys and molecular methods. PCR assays were used to assess Plasmodium infection and mosquito blood meal source.ResultsCatches of adult An. stephensi were generally low (mean: 0.15 per trap), with eight positive sites among the 26 surveyed. This mosquito species was reported for the first time in Assosa, western Ethiopia. Anopheles stephensi was the predominant species in four of the eight positive sites, accounting for 75–100% relative abundance of the adult Anopheles catches. Household-level exposure, defined as the percentage of households with a peridomestic presence of An. stephensi, ranged from 18% in Metehara to 30% in Danan. Anopheles arabiensis was the predominant species in 20 of the 26 sites, accounting for 42.9–100% of the Anopheles catches. Bovine blood index, ovine blood index and human blood index values were 69.2%, 32.3% and 24.6%, respectively, for An. stephensi, and 65.4%, 46.7% and 35.8%, respectively, for An. arabiensis. None of the 197 An. stephensi mosquitoes assayed tested positive for Plasmodium sporozoite, while of the 1434 An. arabiensis mosquitoes assayed, 62 were positive for Plasmodium (10 for P. falciparum and 52 for P. vivax).ConclusionsThis study shows that the geographical range of An. stephensi has expanded to western Ethiopia. Strongly zoophagic behavior coupled with low adult catches might explain the absence of Plasmodium infection. The level of household exposure to An. stephensi in this study varied across positive sites. Further research is needed to better understand the bionomics and contribution of An. stephensi to malaria transmission.Graphical

The synthesis of nanosilver and carbamat to control of Anopheles in malaria endemic areas

Malaria is a national and global problem that really needs attention and Indonesia is the second highest country with the highest number of cases 304,607 in 2021. The use of insecticides at inappropriate doses causes resistance to targets. The aim of this study was to analyze the toxicity of silver nanoparticles to adult Anopheles mosquitoes in malaria endemic areas as well as to determine the lethal concentration (LC50) and to project effectiveness in the lifetime. This research is experimental and uses a post-treatment with control design which was carried out in 4 stages from 2019-2022, which includes laboratory scale tests (2019), field laboratory scale tests (2020), ecological endemic area scale tests (2021), and lifetime tests (2022). Calculation of LC50 in nanosilver toxicity 5965 mg/l and Calculation of life time projections produces the formula Y=(-) 0.012Ln(X)+0.6933. Absorption of silver nanoparticles into the bodies of Anopheles sp. occurs through the spiracles and attacks the Ache enzyme in the central nervous system thereby stopping the working system of the mosquito which will then die. Nanosilver is a very effective herbal insecticide used as an effort to control malaria cases in endemic areas with the right dose and method. For life time, a dose of 1500 mg/l nanosilver effectively kills 74% of adult Anopheles sp mosquitoes on day 32 and it is projected that on 1 year the efficacy of nanomaterials still has an efficacy of 62%).

Anopheles arabiensis continues to be the primary vector of Plasmodium falciparum after decades of malaria control in southwestern Ethiopia

BackgroundInvestigating the species distribution and their role in malaria transmission is important as it varies from place to place and is highly needed to design interventions appropriate to the site. The current study aimed to investigate the Anopheles mosquito species distribution and their infection rate in southwestern Ethiopia.MethodsThe study was conducted in 14 malaria-endemic kebeles (the smallest administrative unit), which were situated in eight different malaria-endemic districts and four zones in southwestern Ethiopia. Ten per cent of households in each village were visited to collect adult mosquitoes using Centers for Disease Control and Prevention (CDC) light traps. The larval and pupal collection was done from breeding sites within the villages, and reared to adults. Female mosquitoes were morphologically identified. The head and thorax of adult Anopheles mosquitoes were tested for circumsporozoite proteins (CSPs) using ELISA. At the same time, legs, wings, and abdomen were used to identify sibling species using PCR targeting the rDNA intergenic spacers region for species typing of the Anopheles funestus group and the internal transcribed spacer 2 region genes for Anopheles gambiae complex.ResultsA total of 1445 Anopheles mosquitoes comprising eight species were collected. Of 813 An. gambiae complex tested by PCR, 785 (97%) were Anopheles arabiensis, and the remaining 28 (3%) were not amplified. There were 133 An. funestus group captured and tested to identify the species, of which 117 (88%) were positive for Anopheles parensis, and 15 (11%) were not amplified. A single specimen (1%) showed a band with a different base pair length from the known An. funestus group species. Sequencing revealed this was Anopheles sergentii. Among 1399 Anopheles tested for CSPs by ELISA, 5 (0.4%) An. arabiensis were positive for Plasmodium falciparum and a single (0.07%) was positive for Plasmodium vivax.ConclusionsAnopheles arabiensis continues to play the principal role in malaria transmission despite implementing indoor-based interventions for decades. Sequencing results suggest that An. sergentii was amplified by the An. funestus group primer, producing PCR amplicon size of different length. Therefore, relying solely on amplifying a specific gene of interest in grouping species could be misleading, as different species may share the same gene.

Label-free visualization of internal organs and assessment of anatomical differences among adult Anopheles, Aedes, and Culex mosquito specimens using bidirectional optical coherence tomography

Label-free visualization of internal organs and assessment of anatomical differences among adult Anopheles, Aedes, and Culex mosquito specimens using bidirectional optical coherence tomography

Mosquito Control Strategies and Insecticide Resistance of the Malaria Vector in Urbanized Land Use Types in Suame Municipality, Ghana.

Modified landscapes could create breeding habitats for mosquitoes and affect their community structure and susceptibility with implications for their management. Hence, in this study, household mosquito control methods in two urbanized landscapes; industrial and residential human settlements, in Ghana and insecticide susceptibility of the inhabiting Anopheles populations were assessed. Household knowledge and usage pattern of mosquito control methods in the modified landscapes were obtained using a questionnaire. Female adult Anopheles mosquitoes were also subjected to susceptibility tests using mosquito coils (0.08% meperfluthrin, 0.03% dimefluthrin, and 0.3% transfluthrin) and World Health Organization (WHO) insecticide test papers (0.05% deltamethrin, 4% DDT, 0.1% bendiocarb, 0.25% pirimiphos-methyl, and 5% malathion). Although insecticide-treated nets and aerosols were used for mosquito control, mosquito coils were the most common and widely preferred household method. The Anopheles mosquitoes were resistant to all the insecticides (mosquito coils and WHO test papers) except pirimiphos-methyl. Land use type did not affect insecticide resistance, but the insecticide type did. The findings indicate the effect of household insecticide usage practices on local mosquito populations and their implications for effective vector management and disease control in modified landscapes.