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Wolbachia pipientis (= Wolbachia) has promise as a tool to suppress virus transmission by Aedes aegypti mosquitoes. However, Wolbachia can have variable effects on mosquito-borne viruses. This variation remains poorly characterized, yet the multimodal effects of Wolbachia on diverse pathogens could have important implications for public health. Here, we examine the effects of transient somatic infection with two strains of Wolbachia (wAlbB and wMel) on the alphaviruses Sindbis virus (SINV), O'nyong-nyong virus (ONNV), and Mayaro virus (MAYV) in Ae. aegypti. We found variable effects of Wolbachia including enhancement and suppression of viral infections, with some effects depending on Wolbachia strain. Both wAlbB- and wMel-infected mosquitoes showed enhancement of SINV infection rates one week post-infection, with wAlbB-infected mosquitoes also having higher viral titers than controls. Infection rates with ONNV were low across all treatments and no significant effects of Wolbachia were observed. The effects of Wolbachia on MAYV infections were strikingly strain-specific; wMel strongly blocked MAYV infections and suppressed viral titers, while wAlbB had more modest effects. The variable effects of Wolbachia on vector competence underscore the importance of further research into how this bacterium impacts the virome of wild mosquitoes including the emergent human pathogens they transmit. Author summary: In recent years, wild populations of Aedes aegypti mosquitoes in over a dozen countries have been deliberately infected with Wolbachia pipientis ("Wolbachia"); an intracellular bacterium that, in some circumstances, helps to curb the spread of mosquito-brone pathogens including dengue virus. But how does Wolbachia affect the ability of mosquitoes to become infected with and spread the many different viruses they encounter in nature? Here, we use transient somatic infections in Aedes aegypti to characterize the effects of Wolbachia on three different alphaviruses that cause illness in humans: Sindbis virus, O'nyong-nyong virus, and Mayaro virus. We find that transient Wolbachia infections have variable effects on these different pathogens, ranging from significant suppression of Mayaro virus to significant enhancement of Sindbis virus. Our research has important implications for the design of vector control strategies, and suggests further research is needed to understand how Wolbachia shapes the replication and transmission of diverse viruses in mosquitoes. [ABSTRACT FROM AUTHOR]

Background: The decision to imbibe a blood meal is predominantly dependent on the sensitivity and specificity of haematophagous arthropods to blood-derived adenine nucleotides, in particular adenosine triphosphate (ATP). Despite previous efforts to identify and characterise the specificity and sensitivity to ATP and other adenine nucleotides, as well as the role of other blood-derived phagostimulants across the Culicidae, comparisons across species remain difficult. Methods: The feeding response of the yellow fever mosquito Aedes aegypti and the African malaria vector Anopheles gambiae to adenine nucleotides in the presence of a carbonate buffer was assessed using a membrane feeding assay. The proportion of mosquitoes engorged and the volume imbibed by all mosquitoes was scored visually and spectrophotometrically. In addition, the proportion of prediuresing An. gambiae, as well as the volume engorged and prediuresed, was examined. Results: Aedes aegypti was more sensitive to adenine nucleotides than An. gambiae, but both species maintained specificity to these phagostimulants, demonstrating a dose-dependent bimodal feeding pattern, thereby expanding our understanding of the all-or-none blood-feeding hypothesis. Feeding on the bicarbonate buffer by An. gambiae—but not that of Ae. aegypti—demonstrated a species-specific variation in how blood phagostimulants are encoded. Adenine nucleotides, with and without bovine serum albumin, were observed to dose-dependently regulate the proportion of An. gambiae prediuresing and the volumes prediuresed but not volumes engorged. Conclusions: Taken together, the results of this study expand our understanding of how mosquitoes differentially assess and respond to blood meal constituents, and provide a basis for further physiological and molecular studies. [ABSTRACT FROM AUTHOR]

Discussions on the Anthropocene have called for increased attention to how the effects of human actions on the planet are accounted for. While much of this debate remains at a theoretical level, more studies on situated Anthropocene realities have been called for. Contributing to the latter, this article explores how experimental and clinical interventions are being accounted for in life science laboratories. Drawing on three research cases, genetically modified mosquitoes, laboratory-grown stem cell lines and assisted reproduction in the IVF clinic, the article traces how 'strategic naturalizing' is conducted to make sense of and justify human interventions on cells, bodies and ecosystems. Strategic naturalizing in laboratory work is situated, fluid, and also implicates the presence of the social scientist. Although naturalizing biotechnologies is not a new phenomenon, according to our observations scientists increasingly draw on notions of a planet profoundly transformed by human intervention as they conceptualize their own accountability. Consequently, we propose that strategic naturalizing is emerging as an elemental logic in the patchy local enactments of the Anthropocene and its concerns. In the context of experimental and clinical laboratory work, it is a key element in enacting good science and orienting the work towards making biotechnology acceptable in wider society. In studying this, the social scientist also is implicated in situated local enactments of the Anthropocene. [ABSTRACT FROM AUTHOR]

The uptake, accumulation, and ontogenetic transfer of microplastics (MPs) in bloodsucking mosquitoes of the genus Anopheles, vectors of helminth and protozoa parasites in humans and animals, were studied under laboratory conditions. In the experiment, 2‐μm‐diameter fluorescent polystyrene spheres were counted in mosquitoes of all stages, from larvae to adults. Larvae from a natural population readily ingested MPs with food, accumulating on average 3.9 × 106 particles per larva within 3 days. The concentration of MPs decreased significantly in mosquitoes during their transition from stage to stage. The average number of detected MPs attained 110 particles per pupa and 3.0 particles per adult. MP uptake by mosquitoes did not affect their survival, whereas the rate of metamorphosis in MP‐consuming mosquitoes was slightly higher. Our data confirmed that MPs can be transferred from feeding larvae to non‐feeding pupae and adult flying Anopheles mosquitoes from natural populations in aquatic ecosystems. [ABSTRACT FROM AUTHOR]

In this paper, we establish and study a discrete mosquito population model with periodic parameters that takes into account the seasonal variation of environments. We are mainly interested in finding a positive periodic solution that is asymptotically stable and attracts all positive solutions. The existence, uniqueness, and stability of periodic solutions of the model are investigated. We show that the instability of the origin implies that the model has a unique asymptotically stable positive periodic solution and attracts all positive solutions and that the local stability of the origin implies its global asymptotic stability. We also give a necessary and sufficient condition for the origin to be stable. Numerical examples are provided to illustrate our theoretical results. [ABSTRACT FROM AUTHOR]

We propose a reaction-diffusion model to describe the interactive wild and sterile mosquito populations in spatially heterogeneous environments, where the vital dynamics of mosquito populations, e.g., offspring produced per mate, the death rates, the amount of releasing sterile mosquitoes, depend on its location habits and the distribution of wild mosquito populations. First, we show the model may present multiple positive steady states with either Neumann or Dirichlet boundary conditions. Then we investigate the interactive dynamics of mosquito populations with the different initial data. Our results show that wild mosquitoes can be eradicated or suppressed within a certain range with different releasing amounts. Finally, numerical simulations further show that the spatial variability environment within the habitat may play a crucial role in extirpating wild mosquito populations in the context of the sterile insect techniques (SIT). [ABSTRACT FROM AUTHOR]

BACKGROUND: Controlling the spread of arboviral diseases remains a considerable challenge due to the rapid development of insecticide resistance in Aedes mosquitoes. This study evaluated the effects of boric acid‐containing toxic sugar bait (TSB) on field populations of resistant Aedes aegypti mosquitoes. In addition, this study examined the flight activity and wing beat frequency and amplitude of males and the flight activity, fecundity, and insemination of females after pairing with males exposed to TSB. The population dynamics of Aedes mosquitoes under imbalanced sex ratios were examined to simulate realistic field conditions for male suppression under the effect of TSB. RESULTS: The mortality of male mosquitoes was consistently high within 24 h after exposure. By contrast, the mortality of female mosquitoes was inconsistent, with over 70% mortality observed at 168 h. The flight activity and wing beat amplitude of treated males were significantly lower than those of controls, but no significant difference in wing beat frequency was detected. The fecundity and insemination of treated female mosquitoes were lower than those of controls. A simulation study indicated that considerably low male population densities led to mating failures, triggering a mate‐finding Allee effect and resulting in persistently low population levels. CONCLUSION: Boric acid‐containing TSB could effectively complement current chemical intervention approaches to control resistant mosquito populations. TSB is effective in reducing field male populations and impairing male flight activity and female‐seeking behavior, resulting in decreased fecundity and insemination. Male suppression due to TSB potentially results in a small mosquito population. © 2024 Society of Chemical Industry. [ABSTRACT FROM AUTHOR]

Background: Mosquitoes serve as vectors for numerous pathogens, posing significant health risks to humans and animals. Understanding the complex interactions within mosquito microbiota is crucial for deciphering vector-pathogen dynamics and developing effective disease management strategies. Here, we investigated the nested patterns of Wolbachia endosymbionts and Escherichia-Shigella within the microbiota of laboratory-reared Culex pipiens f. molestus and Culex quinquefasciatus mosquitoes. We hypothesized that Wolbachia would exhibit a structured pattern reflective of its co-evolved relationship with both mosquito species, while Escherichia-Shigella would display a more dynamic pattern influenced by environmental factors. Results: Our analysis revealed different microbial compositions between the two mosquito species, although some microorganisms were common to both. Network analysis revealed distinct community structures and interaction patterns for these bacteria in the microbiota of each mosquito species. Escherichia-Shigella appeared prominently within major network modules in both mosquito species, particularly in module P4 of Cx. pipiens f. molestus, interacting with 93 nodes, and in module Q3 of Cx. quinquefasciatus, interacting with 161 nodes, sharing 55 nodes across both species. On the other hand, Wolbachia appeared in disparate modules: module P3 in Cx. pipiens f. molestus and a distinct module with a single additional taxon in Cx. quinquefasciatus, showing species-specific interactions and no shared taxa. Through computer simulations, we evaluated how the removal of Wolbachia or Escherichia-Shigella affects network robustness. In Cx. pipiens f. molestus, removal of Wolbachia led to a decrease in network connectivity, while Escherichia-Shigella removal had a minimal impact. Conversely, in Cx. quinquefasciatus, removal of Escherichia-Shigella resulted in decreased network stability, whereas Wolbachia removal had minimal effect. Conclusions: Contrary to our hypothesis, the findings indicate that Wolbachia displays a more dynamic pattern of associations within the microbiota of Culex pipiens f. molestus and Culex quinquefasciatus mosquitoes, than Escherichia-Shigella. The differential effects on network robustness upon Wolbachia or Escherichia-Shigella removal suggest that these bacteria play distinct roles in maintaining community stability within the microbiota of the two mosquito species. [ABSTRACT FROM AUTHOR]

Anophelinae mosquitoes are exposed to a variety of microbes including Plasmodium parasites that cause malaria. When infected, mosquitoes mount versatile immune responses, including the production of antimicrobial peptides. Cecropins are one of the most widely distributed families of antimicrobial peptides in insects and all previously studied Anopheles members are playing roles in adult mosquito immunity. We have identified and characterized a novel member of the Anopheles gambiae cecropin family, cecropin D (CecD), that is uniquely expressed and immune-responsive at late larval stages to promote microbial clearance through its broad-spectrum antibacterial activity during larval-pupal developmental transition. Interestingly, Cecropin D also exhibited highly potent activity against Plasmodium falciparum sporozoites, the malaria parasite stage that is transmitted from mosquitoes and infects humans and thereby holds promise as a malaria transmission-blocking agent. Finally, we have defined unequivocal cecropin-specific molecular signatures to systematically organize the diversity of the cecropin family in malaria vectors. Author summary: Anopheles mosquitoes that transmit the deadly malaria are exposed to a variety of microbes in their natural habitats. Mosquitoes use their innate immune system that also comprises antimicrobial peptides to fight infections with these microbes including the Plasmodium malaria parasite. Cecropins are one of the most widely distributed antimicrobial peptides in insects and all previously studied Anopheles cecropins are playing roles in adult mosquito immunity. We have identified and characterized a novel Anopheles gambiae cecropin, cecropin D (CecD), that is uniquely produced and immune-responsive at late larval stages to promote broad spectrum microbial clearance during larval-pupal developmental transition. Interestingly, Cecropin D also shows potent activity against Plasmodium falciparum sporozoites, the malaria parasite stage that is transmitted from mosquitoes to humans and could therefore be developed into a malaria control strategy. [ABSTRACT FROM AUTHOR]

This study delves into the often‐overlooked impact of immature mosquitoes on the dynamics of malaria transmission. By employing a mathematical model, we explore how these aquatic stages of the vector shape the spread of the disease. Our analytical findings are corroborated through numerical simulations conducted using the Runge–Kutta fourth‐order method in MATLAB. Our research highlights a critical factor in malaria epidemiology: the basic reproduction number R0. We demonstrate that when R0 is below unity R0<1, the disease‐free equilibrium exhibits local asymptotic stability. Conversely, when R0 surpasses unity R0>1, the disease‐free equilibrium becomes unstable, potentially resulting in sustained malaria transmission. Furthermore, our analysis covers equilibrium points, stability assessments, bifurcation phenomena, and sensitivity analyses. These insights shed light on essential aspects of malaria control strategies, offering valuable guidance for effective intervention measures. [ABSTRACT FROM AUTHOR]

Background: Despite 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). Results: There 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. Conclusions: The 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. [ABSTRACT FROM AUTHOR]

Background: Study of malaria parasite cell biology is challenged by their small size, which can make visualisation of individual organelles difficult or impossible using conventional light microscopy. In recent years, the field has attempted to overcome this challenge through the application of ultrastructure expansion microscopy (U-ExM), which physically expands a biological sample approximately 4.5-fold. To date, U-ExM has mostly been used to visualise blood-stage parasites and used exclusively on parasites in vitro. Methods: Here we develop Mosquito Tissue U-ExM (MoTissU-ExM), a method for preparing dissected mosquito salivary glands and midguts by U-ExM. MoTissU-ExM preserves both host and parasite ultrastructure, enabling visualisation of oocysts and sporozoites in situ. We also provide a point-by-point protocol for how to perform MoTissU-ExM. Results: We validate that MoTissU-ExM samples expand as expected, provide a direct comparison of the same dissected tissues before and after MoTissU-ExM, and highlight some of the key host and parasite structures that can be visualised following MoTissU-ExM. Discussion: We discuss potential use cases for MoTissU-ExM for study of malaria parasite biology, and more broadly. We detail drawbacks or challenges MoTissU-ExM and imaging these expanded tissues, along with information troubleshooting this technique. Finally, we discuss how MoTissU-ExM could be applied and adapted in future to increase its utility. [ABSTRACT FROM AUTHOR]

Accurate species identification of the mosquitoes in the genus Anopheles is of crucial importance to implement malaria control measures and monitor their effectiveness. We use a previously developed amplicon panel (ANOSPP) that retrieves sequence data from multiple short nuclear loci for any species in the genus. Species assignment is based on comparison of samples to a reference index using k-mer distance. Here, we provide a protocol to generate version controlled updates of the reference index and present its latest release, NNv2, which contains 91 species, compared to 56 species represented in its predecessor NNv1. With the updated reference index, we are able to assign samples to species level that previously could not be assigned. We discuss what happens if a species is not represented in the reference index and how this can be addressed in a future update. To demonstrate the increased power of NNv2, we showcase the assignments of 1789 wild-caught mosquitoes from Madagascar and demonstrate that we can detect within species population structure from the amplicon sequencing data. [ABSTRACT FROM AUTHOR]

Background: Plasmodium knowlesi, identified as the fifth human malaria parasite, has rapidly spread across various Southeast Asian countries, yet uncertainties persist regarding its human-mosquito-human transmission. Therefore, this study aims to explore the transmission potential of P. knowlesi from human blood to mosquitoes. Methods: A direct membrane-feeding assay was conducted by infecting laboratory-reared female Anopheles dirus mosquitoes with P. knowlesi-infected human blood from a single patient presenting with febrile malaria. Mosquitoes were dissected 7 days post-infection under a stereomicroscope to detect oocysts in the midgut, stained with mercurochrome. Salivary glands were examined 14 days post-infection for the presence of sporozoites. Malaria diagnosis employed microscopy by expert microscopists and nested PCR assays. Results: Upon dissecting 745 out of 1439 blood-fed An. dirus mosquitoes on day 7 post-infection, two oocysts were identified in the midguts of two mosquitoes (0.27%). An additional 694 mosquitoes were dissected for salivary glands on day 14 post-infection, with three mosquitoes (0.43%) exhibiting sporozoites. Further confirmation by nested-PCR assay verified these sporozoites as belonging to the P. knowlesi species. Conclusions: The findings underscore the potential transmission of P. knowlesi from human blood to mosquitoes. The significance of these findings necessitates further investigation, such as repeating similar experiments among natural vectors, to gain deeper insights into the transmission dynamics of P. knowlesi in Southeast Asia. [ABSTRACT FROM AUTHOR]

The resting and feeding behaviour of the main malaria vectors was studied in rural areas of Côte d'Ivoire, after Long-Lasting Insecticidal Nets (LLINs) national coverage. Adult mosquitoes were sampled by Human Landing Catches (HLC) and Pyrethrum Spray Catches (PSC). The annual entomological inoculation rate (EIR) was calculated for each species. An. coluzzii, An. gambiae s.s. and An. funestus were the vector species identified in Toumbokro while An. coluzzii, An. funestus and An. nili were identified in Kassiapleu. These vectors feed preferentially on humans in the two villages. In Toumbokro, An. gambiae s.l. is exophagic while An. funestus is endophagic. In this village, the annual Entomological Inoculation Rate (EIR) average was high and estimated at 228.49 infectious bites per person per year (ib/p/y) with the contribution 80.81 ib/p/y either 35% for An. gambiae s.l. and 147.68 ib/p/y either 65% for An. funestus. An. funestus has become the main vector and provide its own more than half of this transmission. In Kassiapleu, this EIR was also high and estimated at 348.58 ib/p/y. An. gambiae s.l. and An. funestus have been responsible for 332.15 ib/p/y either 95% and 16.43 ib/p/y either 5%, respectively. An. gambiae s.l. is endophagic and has remained the main vector of malaria. This work shows that malaria transmission remains high and is assured by two main vectors (An. funestus and An. gambiae s.l.) in rural areas of Côte d'Ivoire. These vectors bite both inside and outside the house. Thus, effective malaria control requires combining indoor residual spraying and outdoor control tools at LLINs currently used against the vectors inside the home. [ABSTRACT FROM AUTHOR]

Background: It is widely reported that exotic larvicidal fishes like Gambusia and Poecilia have adverse impact on the native aquatic fauna. The present study highlights the efficacy of two colourful native murrels, primarily designated as ornamental fish, Channa bleheri and Channa stewartii as biocontrol agent of mosquito larvae. Methods: Live specimens of Channa bleheri and C. stewartii, collected from the wetlands of Tinsukia district of Upper Assam, were assessed for their larvivorous potential at individual and group levels during day and night by dividing the specimens into two size groups. After 12 hour and 24 hour starvation, the test specimens (mean size for small group 8.67-9.17 cm and that of large group 11.63-13.27 cm) were given known number of mosquito larvae and recorded the consumption rate. Result: The predation rate varied from 33.3±4.36 to 71.6±5.15/min for Channa bleheri and that of C. stewartii from 16.3±0.95 to 68.2±2.77/min. In both species, smaller sized specimens were better performers as predators. Predation rate at 12 and 24 hrs of starvation and between day and night proved that these native murrels are excellent predators of mosquito larvae. [ABSTRACT FROM AUTHOR]

This study presented the first sequencing and analysis of the mitochondrial genome of Paraclius curvispinus Yang and Saigusa, 2001, a predator fly belonging to the family Dolichopodidae (long-legged fly). The complete mitochondrial genome of P. curvispinus was 16,882 bp long (GenBank accession number: PP498802), which contained 13 protein-coding genes, 22 tRNAs, 2 rRNAs, and a control region. The nucleotide composition of mitochondrial genome of P. curvispinus was 38.48% of A, 36.00% of T, 10.67% of G, and 14.85% of C. Maximum-likelihood (ML) analysis revealed the phylogenetic relationships within the subfamily Dolichopodinae, suggesting the following grouping: ((Dolichopus + Lichtwardtia)+((Hercostomus + Poecilobothrus)+ Gymnopternus))+ Paraclius. This study provides a basis for discussing the evolution of mosquito larval predation habits of Dolichopodinae. [ABSTRACT FROM AUTHOR]

To gain insights into the effectiveness of the newly proposed and developed genetic sterile insect technique in control of mosquitoes, that releases mosquitoes carrying a dominant lethal (RIDL), we formulate discrete-time models based on difference equations. We incorporate, respectively, the four release strategies, early acting bisex (EBS), late acting bisex (LBS), early acting female-killing (EFK), and late acting female-killing (EFK) lethality in the models. We investigate the existence and stability of positive fixed points and derive a release threshold of sterile mosquitoes for each model. We compare the different release strategies by numerical examples. [ABSTRACT FROM AUTHOR]

Mosquito‐borne viruses continue to affect billions of people globally, posing a severe health risk and an economic burden. Aedes albopictus (Skuse), a highly invasive mosquito species, has repeatedly invaded and increased its presence, serving as a key vector of dengue virus, yellow fever virus (YFV), Zika virus (ZIKV), and Chikungunya virus (CHIKV), causing frequent outbreaks of related viral diseases. This study investigated the impact of larval diet quantity on larval duration and adult body size. The effect of adult mosquito body size on various aspects of Ae. albopictus was also examined, including blood‐feeding behavior, follicular development, reproductive capacity, egg retention capacity, preoviposition period, and fecundity. These diverse characteristics all have an effect on arboviruses transmission. The changes in body size (small, medium, and large) are obtained by providing different quantities of larval diet (low, average, and high). The results indicate that the quantity of larval diet directly impacts the adult body size while inversely affecting the larval duration. Furthermore, a positive correlation exists between adult body size and wing length, implying that wing length could be a reliable indicator of adult body size and rearing conditions during the developmental stages. Large females exhibited higher numbers of follicles and greater fecundity. Moreover, a significant correlation was observed between follicle number before the first blood meal and total egg number. In contrast, increasing wing length decreased the number of blood meals, egg retention, and the preoviposition period. The tendency of small females to perform multiple feedings was greater than that of large females. Small females exhibited a higher propensity for multiple feeding activities when compared to their larger counterparts. Most medium‐sized females (92.8%) deposited eggs in their ovaries, however, 7.2% retained a few. In contrast, most large females (87.4%) had complete ovary egg‐laying, whereas a minority (12.6%) retained some of their eggs. About 35.2% of small females showed ovarian egg retention, while 64.8% successfully laid all their eggs. After the first blood meal, the oviposition rate was 92% for large females, 88% for medium females, and 76% for small females. About 69.86% of the follicles in large females underwent vitellogenesis. This finding suggests that small females with low energy reserves exhibited incomplete oviposition and multiple blood feedings to increase their reproductive capacity. Research Highlights: Add more information about arbovirus epidemics and their consequences.Aedes albopictus is a global invasive species that transmit dengue virus, CHIKV, YFV, and ZIKV.A negative correlation was observed between body size, egg retention, and multiple blood feedings in Aedes albopictus.Size of the female's body was positively correlated with fecundity, while it was negatively correlated with the preoviposition period.Size‐dependent multiple blood feeding affects vector‐host contact frequency. [ABSTRACT FROM AUTHOR]

Notonecta species (Hemiptera: Notonectidae) are generalist true predators with a strong impact on the size, structure, and richness of the aquatic communities, particularly on mosquito populations. Juvenile and adult backswimmers not only prey on mosquito larvae (consumptive effect) but their presence can also deter the oviposition of gravid mosquito females (non‐consumptive effect). In this study, we selected a Western Palearctic backswimmer, Notonecta maculata Fabricius, as a predator and deterrent of the highly invasive and health‐threatening Asian tiger mosquito, Aedes albopictus (Skuse) (Diptera: Culicidae). In central Italy, N. maculata is univoltine and overwinters in the adult stage. Females lay eggs from February to the end of May, and the first new adults will emerge in late June; the complete nymphal development requires 44.5 ± 3.1 days. The number of Ae. albopictus larvae preyed on by N. maculata during its growth increases while developing, going from 0.03 ± 0.02 larvae per day for first instar nymphs to 21.38 ± 2.90 for mature adults. Furthermore, the presence of a single N. maculata adult in a mosquito ovitrap is enough to discourage the Ae. albopictus oviposition for up to 5 days. Limiting mosquito populations and, consequently, the transmission of viruses and parasites should be our main goal. Therefore, the integrated management of Ae. albopictus by the use of insecticides and a predator (such as backswimmer juveniles and adults), even if only in specific situations, may be promising and feasible. A more in‐depth knowledge of the biological cycle of N. maculata could help, consequently, to optimize its mass rearing. [ABSTRACT FROM AUTHOR]

We report a convergent synthesis of heteroaryl/aryl-annulated pyridine and quinoline derivatives by a metal-free Povarov reaction. para -Toluene sulfonic acid was used as the catalyst in this reaction, which produced the products in good yields from the corresponding aromatic amines and ethyl vinyl ether. A pyridocoumarin and a pyridopyrimidine product were evaluated for their mosquito larvicidal activity against the third instar larvae of the dengue vector mosquito Aedes aegypti. Examination of morphological changes in the larvae showed damage to the target body part after treatment with both the pyridocoumarin and pyridopyrimidine products at the LC50 concentrations. [ABSTRACT FROM AUTHOR]

It is commonly recognized that predation and the risk of predation can have profound effects on behaviors and population ecology of prey. Comparatively, little attention has been paid to the trade-offs hosts make to manage the risk of parasitism, but there is evidence that hosts make behavioral trade-offs to avoid parasitism in ways that resemble those made by prey animals under the risk of predation. Mosquitoes are common hematophagous ectoparasites which parasitize a wide range of vertebrates, resulting in blood loss, distraction, and disease transmission. Many hosts engage in defensive behaviors to repel or kill mosquitoes and these behaviors likely come at the expense of activities such as foraging and vigilance for predators. We placed a mosquito attractant (BG-Sweetscent) and repellant (allethrin) near artificial foraging patches in areas where mosquitoes were common and measured the amount of grain left after a night of foraging (giving-up density) to investigate whether mosquitoes influenced Cotton Mouse (Peromyscus gossypinus) foraging. We predicted that in patches with mosquito attractants, mosquito harassment would cause mice to engage in defensive behaviors which would distract or deter them from foraging, resulting in less grain consumption relative to control patches. Conversely, we predicted that in mosquito-repellent patches, mice would experience less harassment, resulting in more grain consumption. Indoor arena trials found no evidence that the mosquito treatments influenced cotton mouse behavior in the absence of mosquitoes. However, we found no evidence that mosquito treatments affected giving-up densities. [ABSTRACT FROM AUTHOR]

Phylogenetic analyses showed that the virus responsible for a May 2024 Oropouche fever outbreak in Cuba was closely related to viruses from Brazil in 2023. Pools of Ceratopogonidae spp. biting midges and Culex quinquefasciatus mosquitoes were positive for Oropouche viral RNA. No cases were severe. Virus extension to new areas may increase case numbers and severity [ABSTRACT FROM AUTHOR]

Orthoflaviviruses are arthropod-borne viruses that are transmitted by mosquitoes or ticks and cause a range of significant human diseases. Among the most important tick-borne orthoflaviviruses (TBFVs) is tick-borne encephalitis virus (TBEV), which is endemic in Eurasia, and Powassan virus, which is endemic in Asia and North America. There is a significant controversy regarding species assignment in the tick-borne encephalitis virus complex due to the complex phylogenetic, serological, ecological, and pathogenetic properties of viruses. Comparing the rate of non-synonymous to synonymous substitutions (dN/dS) over the course of tick-borne orthoflavivirus diversification suggests that there is a very strong stabilizing selection (Nei-Gojobori dN/dS < 0.1) among tick-borne orthoflaviviruses that differ by less than 13.5% amino acid/21.4% nucleotide sequences, and discretely more rapid accumulation of non-synonymous substitutions (dN/dS > 0.13) among more divergent viruses that belong to distinct species. This pattern was similarly observed in genome regions encoding structural (E) and non-structural (NS3) proteins. Below this distance threshold, viruses appear fit and strongly tied to their ecological niche, whereas above the threshold, a greater degree of adaptation appears necessary. This species criterion suggests that all subtypes of TBEV, all related ovine/caprine encephalomyelitis viruses, and Omsk hemorrhagic fever virus (OHFV) together correspond to a single species. Within this species, viruses make up 11 subtypes that are reliably segregated by a 10% nucleotide distance cut-off suggested earlier for TBEV. The same 10% subtype cut-off suggests that Powassan virus includes two subtypes, Powassan and Deer Tick virus. [ABSTRACT FROM AUTHOR]

The RNA genome of orthoflaviviruses encodes a methyltransferase within the non-structural protein NS5, which is involved in 2'-O-methylation of the 5'-terminal nucleotide of the viral genome resulting in a cap1 structure. While a 2'-O-unmethylated cap0 structure is recognized in vertebrates by the RNA sensor RIG-I, the cap1 structure allows orthoflaviviruses to evade the vertebrate innate immune system. Here, we analyzed whether the cap0 structure is also recognized in mosquitoes. Replication analyses of 2'-O-methyltransferase deficient yellow fever virus mutants (YFV NS5-E218A) of the vaccine 17D and the wild-type Asibi strain in mosquito cells revealed a distinct downregulation of the cap0 viruses. Interestingly, the level of inhibition differed for various mosquito cells. The most striking difference was found in Aedes albopictus-derived C6/36 cells with YFV-17D cap0 replication being completely blocked. Replication of YFV-Asibi cap0 was also suppressed in mosquito cells but to a lower extent. Analyses using chimeras between YFV-17D and YFV-Asibi suggest that a synergistic effect of several mutations across the viral genome accompanied by a faster initial growth rate of YFV-Asibi cap1 correlates with the lower level of YFV-Asibi cap0 attenuation. Viral growth analyses in Dicer-2 knockout cells demonstrated that Dicer-2 is entirely dispensable for attenuating the YFV cap0 viruses. Translation of a replication-incompetent cap0 reporter YFV-17D genome was reduced in mosquito cells, indicating a cap0 sensing translation regulation mechanism. Further, oral infection of Aedes aegypti mosquitoes resulted in lower infection rates for YFV-Asibi cap0. The latter is related to lower viral loads found in the midguts, which largely diminished dissemination to secondary tissues. After intrathoracic infection, YFV-Asibi cap0 replicated slower and to decreased amounts in secondary tissues compared to YFV-Asibi cap1. These results suggest the existence of an ubiquitously expressed innate antiviral protein recognizing 5'-terminal RNA cap-modifications in mosquitoes, both in the midgut as well as in secondary tissues. Author summary: Arthropod-borne viruses infect both mosquito and vertebrate hosts. One factor influencing the level of viral replication in the host is the ability of the virus to evade or counteract the host's immune system. In vertebrates, it was shown that orthoflaviviruses circumvent the innate immune system by capping their viral genome with a cap1 structure characterized by 2'-O-methylation of the 5'-terminal nucleotide. In contrast, viral genomes lacking 2'-O-methylation (cap0) are recognized as foreign and activate the vertebrate innate immune system. Here, we used yellow fever virus (YFV) strain 17D and Asibi mutants lacking 2'-O-methylation, as well as a cap0 reporter YFV genome, to analyze cap0 recognition in mosquito cells and living mosquitoes. The replication of the YFV cap0 mutants was profoundly impaired in different mosquito cells and cap0 YFV genomes showed reduced translation levels. Based on viral infection experiments in Aedes aegypti mosquitoes, we further demonstrate that these vector mosquitoes recognize the cap0 structure and that a potential recognizing effector protein is localized in the midgut as well as in secondary tissues. Our findings provide the basis for future studies investigating the recognition of foreign RNAs in mosquitoes and should contribute to establish methods to prevent transmission of arboviruses. [ABSTRACT FROM AUTHOR]

The aim of the present study was to evaluate the chemical composition of essential oils from Myroxylon peruiferum and Eugenia gracillima and determine the larvicidal potential of the oils against Aedes aegypti. Essential oils from the leaves and young stems of M. peruiferum (EOLMP and EOAMP) and the leaves of E. gracillima (EOLEG) were obtained through hydrodistillation. The major constituents of EOLMP were α-pinene (18.0%) and δ-cadinene (14.2%); the major constituents of EOAMP were spathulenol (22.9%) and β-elemene (12.4%) and the major constituents of EOLEG were δ-elemene (40.4%) and germacrene D (25.2%). The two oils extracted from M. peruiferum were very active against A. aegypti larvae and had similar lethal concentrations (LC50 = 0.7 mg/mL). The oil extracted from E. gracillima had an LC50 of 1.51 mg/mL. The essential oils from M. peruiferum and E. gracillima constitute promising sources for the formulation of novel insecticides for the control of the mosquito Aedes aegypti. [ABSTRACT FROM AUTHOR]

Simple Summary: Biting hematophagous dipterans are responsible for painful bites and blood spoliation; they induce behavioral modifications, anemia, and significant production losses in cattle. A feedlot of 100 feeder cattle would register a total loss of USD 16,000 within 5 months, which appears to be an unexpectedly huge loss caused by dipterans. Investing part of this money in fly control would probably be beneficial. In addition to blood pathogen transmission, insects of the order Diptera affect livestock through visual and contact harassment; blood-feeders are responsible for painful bites and blood despoliation, generating behavioral modifications, anemia, and production losses. Knowledge of their economic impact is a basis for cost-effective control. Here, we measured the global impact of diptera insects by comparing two batches of six feeder cattle, one in the open air and the other protected by a mosquito net. The analytical data were insect density in the open air and, for feeder cattle, tail flick counts, hematocrit values (Ht), feed intake, feed conversion ratio (FCR), and live body weight gain (LBWG). Over a period of five months, the results showed significant losses in the LBWG of cattle exposed to insects, estimated at 8.0 ± 1.5 kg/month [2.7; 13.3], with a total loss reaching 40.0 ± 5.5 kg/head. Main diurnal insects were Stomoxys spp. and Musca crassirostris. There was a strong correlation between fly density and diurnal tail flicks. Night trapping and tail flicks showed a potentially important role of mosquitoes to be further explored. The Ht levels of exposed animals were 3–4% lower than those of controls. FCRs indicated that exposed animals needed 33% more dry matter intake/kg of LBWG. An economic assessment showed that dipterans were responsible for a 10–11% loss in LBWG during the main growing period of feeder cattle (10–15 months). A feedlot of 100 calves would register a total loss of USD 16,000 within 5 months, which appears to be an unexpectedly huge loss caused by dipterans. Investing part of this money into fly control would probably be beneficial. [ABSTRACT FROM AUTHOR]

Background: Surveillance of the host–anopheline mosquitoes' interaction is important for assessing malaria transmission risk and guiding vector control. We assume that changes in malaria vector species' feeding habits, as well as the surrounding environment, have a substantial impact on varied malaria transmission. In this study, we determined the vertebrate host feeding patterns of anopheline mosquitoes to characterize entomologic risk factors for malaria in Jabi Tehnan, Northwestern Ethiopia. Methods: Blood-fed anophelines surveyed during malaria surveillance in Jabi Tehnan district of northwestern Ethiopia were utilized in this study. They were collected using Centers for Disease Control and Prevention (CDC) light traps deployed in selected households per village, placed indoors and outdoors, spanning three agroecological settings (dry mountain, plateau, and semiarid highlands) between June 2020 and May 2021. The engorged mosquitoes were analyzed for host blood meal sources and Plasmodium infection via polymerase chain reaction (PCR) and/or sequencing. Infection rates and bovine and human blood indices were calculated and compared for abundant species; between indoors and outdoors and between agroecology using a chi-squared test for equality of proportion in R package at a significant level of p ≤ 0.05. Results: A total of 246 mosquitoes were successfully typed (indoor, 121; outdoor, 125), with greater relative abundance indoors in mountain and plateau highlands, and outdoors in semiarid areas. Despite ecological differences in blood-fed capture rates, cattle served as the most utilized blood meal source by 11 anopheline species with an overall bovine blood index (BBI) of 74.4%. This trend was dictated by Anopheles gambiae s.l. (198/246; BBI = 73.7%), which exhibited the most plastic feeding habits that included humans (human blood index = 15.7%) and other livestock and rodents. A total of five anopheline species (An. gambiae s.l., An. funestus s.l., An. coustani s.l., An. pretoriensis, and An. pharoensis) fed on humans, of which the first three were found infected with Plasmodium parasites. Most of the infected specimens were An. arabiensis (5.6%, 11/198) and had recently fed mainly on cattle (72.7%, 8/11); one each of infected An. funestus s.l. and An. coustani s.l. had fed on humans and cattle, respectively. Conclusions: The results demonstrate communal feeding on cattle by anophelines including primary and secondary malaria vectors. This study also indicates the importance of cattle-targeted interventions for sustainable control of malaria vectors in the study areas. [ABSTRACT FROM AUTHOR]

Background: Disease-vector mosquito monitoring is an essential prerequisite to optimize control interventions and evidence-based risk predictions. However, conventional entomological monitoring methods are labor- and time-consuming and do not allow high temporal/spatial resolution. In 2022, a novel system coupling an optical sensor with machine learning technologies (VECTRACK) proved effective in counting and identifying Aedes albopictus and Culex pipiens adult females and males. Here, we carried out the first extensive field evaluation of the VECTRACK system to assess: (i) whether the catching capacity of a commercial BG-Mosquitaire trap (BGM) for adult mosquito equipped with VECTRACK (BGM + VECT) was affected by the sensor; (ii) the accuracy of the VECTRACK algorithm in correctly classifying the target mosquito species genus and sex; (iii) Ae. albopictus capture rate of BGM with or without VECTRACK. Methods: The same experimental design was implemented in four areas in northern (Bergamo and Padua districts), central (Rome) and southern (Procida Island, Naples) Italy. In each area, three types of traps—one BGM, one BGM + VECT and the combination of four sticky traps (STs)—were rotated each 48 h in three different sites. Each sampling scheme was replicated three times/area. Collected mosquitoes were counted and identified by both the VECTRACK algorithm and operator-mediated morphological examination. The performance of the VECTRACK system was assessed by generalized linear mixed and linear regression models. Aedes albopictus capture rates of BGMs were calculated based on the known capture rate of ST. Results: A total of 3829 mosquitoes (90.2% Ae. albopictus) were captured in 18 collection-days/trap/site. BGM and BGM + VECT showed a similar performance in collecting target mosquitoes. Results show high correlation between visual and automatic identification methods (Spearman Ae. albopictus: females = 0.97; males = 0.89; P < 0.0001) and low count errors. Moreover, the results allowed quantifying the heterogeneous effectiveness associated with different trap types in collecting Ae. albopictus and predicting estimates of its absolute density. Conclusions: Obtained results strongly support the VECTRACK system as a powerful tool for mosquito monitoring and research, and its applicability over a range of ecological conditions, accounting for its high potential for continuous monitoring with minimal human effort. [ABSTRACT FROM AUTHOR]

Mosquitoes are significant vectors of deadly diseases such as malaria and dengue, contributing to approximately 300,000 deaths annually, according to the World Health Organization (WHO). Traditional chemical repellents, while effective, often cause severe allergic reactions and contribute to mosquito resistance. This study explores the formulation of GAP-F essential oil-loaded microspheres as a natural and eco-friendly alternative for mosquito repellency. Essential oils have demonstrated insect-repellent properties; however, their volatility limits their effectiveness. Microencapsulation is employed to protect these oils from degradation and facilitate sustained release, enhancing their efficacy against mosquitoes. GAP-F essential oil was sourced from the CSIR-CIMAP laboratory through distillation. Laboratory tests were conducted to evaluate its effectiveness against Aedes aegypti, Anopheles stephensi and wild mosquitoes. At a concentration of 10%, GAP-F essential oil exhibited repellency rates of 70%, 85% and 75% against these species during a one-hour bioassay. Given the volatile nature of essential oils, the formulation of microspheres serves as a protective carrier that enhances stability and prolongs the duration of protection. The essential oil-loaded microspheres demonstrated satisfactory results in terms of repellency, showing effectiveness rates of 56.66%, 51.66% and 55% against Aedes aegypti, Anopheles stephensi and wild mosquitoes, respectively, during the same duration. Mosquitoes aged between three to five days were used for the bioassay. This research highlights the potential of microencapsulated natural repellents as a viable alternative to chemical products in mosquito control strategies, providing an effective solution that minimizes health risks while addressing the growing concern of insecticide resistance in mosquito populations. [ABSTRACT FROM AUTHOR]

Mosquitoes serve as vectors for many arthropod-borne viruses (arboviruses) that are responsible for millions of human infections and thousands of deaths each year. Among these arboviruses, O'nyong-nyong virus (ONNV) is an African alphavirus mainly transmitted by Anopheles mosquitoes. ONNV can be detected through serological or molecular tests, the first showing cross-reactivity to co-circulating alphaviruses and requiring technically demanding confirmation, while the latter, usually based on real-time PCR, are costly and demand specific equipment. Isothermal amplification approaches, such as Loop-Mediated Isothermal Amplification (LAMP), should therefore provide a cost-effective, sensitive, and specific alternative for virus detection, suitable for the resource-limited regions where ONNV circulates up to the present time. Here, we describe the development and optimization of a rapid and highly sensitive (10 pfu/reaction) RT-LAMP assay for ONNV detection. Additionally, we demonstrate that it is possible to bypass the RNA extraction step, reducing sample handling time and costs. The final RT-LAMPONNV is a promising field detection tool for ONNV, enabling a better understanding of its impact and serving as a point-of-care diagnostic method. [ABSTRACT FROM AUTHOR]

Malaria is a major international public health problem. The risk of acquiring malaria varies depending on the intensity of transmission and adherence to mosquito precautions and prophylaxis recommendations. Severe malaria can cause significant multiorgan dysfunction, including acute kidney injury (AKI). Intravenous artesunate is the treatment of choice for severe malaria in non-endemic areas. One of the possible events connected with the lifesaving effects of artemisins is post-artesunate haemolysis (PADH), which may be potentially dangerous and under-recognised. We present a case of a seafarer with severe Plasmodium falciparum malaria complicated with AKI and PADH, with a good response to steroid treatment. This case highlights the need for malaria prophylaxis in business travellers, e.g., seafarers to malara-endemic regions, and close supervision of patients with malaria even after the completion of antimalarial treatment due to the possibility of late complications. [ABSTRACT FROM AUTHOR]

West Nile virus (WNV) is a mosquito-borne pathogen with a worldwide distribution. Climate change and human activities have driven the expansion of WNV into new territories in Europe during the last two decades. Romania is endemic for WNV circulation since at least 1996 when the presence of lineage 1 was documented during an unprecedented outbreak. Lineage 2 was first identified in this country during a second significant human outbreak in 2010. Its continuous circulation is marked by clade replacement, and even co-circulation of different strains of the same clade was observed until 2016. The present study aims to fill the information gap regarding the WNV strains that were circulating in Romania between 2017 and 2023, providing chiefly viral sequences obtained from mosquito samples collected in the Bucharest metropolitan area, complemented by human and bird viral sequences. WNV was detected mainly in Culex pipiens mosquitoes, the vectors of this virus in the region, but also in the invasive Aedes albopictus mosquito species. Lineage 2 WNV was identified in mosquito samples collected between 2017 and 2023, as well as in human sera from patients in southern and central Romania during the outbreaks of 2017 and 2018. Both 2a and 2b sub-lineages were identified, with evidence of multiple clusters and sub-clusters within sub-lineage 2a, highlighting the complex and dynamic circulation of WNV in Romania, as a consequence of distinct introduction events from neighboring countries followed by in situ evolution. [ABSTRACT FROM AUTHOR]

The global distribution of Aedes aegypti mosquitoes, particularly in tropical regions, poses a significant public health risk due to their apparent ability to transmit arboviruses such as West Nile virus (WNV). This study aimed to evaluate the vector competence of Ae. aegypti from São Tomé and Príncipe (STP) for the transmission of the WNV PT6.39 strain, considering its potential role as a bridge vector in a region where Culex quinquefasciatus would be the main vector. Aedes aegypti mosquitoes were collected, reared, and experimentally infected with WNV, with viral dissemination and transmission potential assessed 7, 14, and 21 days post infection (dpi). The results showed an increasing trend in infection rates, from 5% at 7 dpi to 35% at 21 dpi, with corresponding dissemination rates of 0%, 100%, and 43%. The transmission rates also increased from 0% at 7 dpi to 67% at 21 dpi, with a maximum transmission efficiency of 10% observed at the final time point. Although Ae. aegypti from STP demonstrated the potential to transmit WNV, the overall transmission efficiency remained relatively low. These findings provide necessary insights into the vector competence of Ae. aegypti in this region, highlighting the importance of continued monitoring and targeted vector control measures to mitigate the risk of potential WNV outbreaks. [ABSTRACT FROM AUTHOR]

Wolbachia are the most spread bacterial endosymbionts in the world. These bacteria can manipulate host reproduction or block virus transmission in mosquitoes. For this reason, Wolbachia-based strategies for vector control are seriously considered or have already been applied in several countries around the world. In South America, Wolbachia have been studied in human pathogen vectors such as sand flies and mosquitoes. In French Guiana, the diversity and distribution of Wolbachia are not well known in mosquitoes. In this study, we screened for Wolbachia natural infection in mosquitoes in French Guiana by using 16S rRNA, Wolbachia surface protein (WSP), and multi-locus sequence typing (MLST) molecular assays. A total of 29 out of 44 (65.9%) mosquito species were positive for natural Wolbachia infection according to the PCR results, and two Wolbachia strains co-infected three specimens of Mansonia titillans. Then, we analyzed the phylogenetic relationships among the Wolbachia detected. All of the tested specimens of Aedes aegypti, the major dengue vector of French Guiana, were negative. These results regarding Wolbachia strain, distribution, and prevalence in mosquitoes from French Guiana highlight Wolbachia–mosquito associations and pave the way for a future Wolbachia-based strategy for vector control in this Amazonian territory. [ABSTRACT FROM AUTHOR]

BACKGROUND: Vector mosquito control is important for preventing and controlling mosquito‐borne infectious diseases. This study designed and developed a mosquito killer (MK) with a specific light wavelength, simulated human body temperature, human odor, and a photocatalyst to stimulate CO2 based on the physiological characteristics and ecological habits of mosquitoes. We tested the trapping effect of individual and multiple mosquito‐trapping elements of the MK through two‐way selection experiments and compared them with several commercial mosquito traps. RESULTS: The 365 nm wavelength MK was significantly more effective than the 395 nm (Cx. quinquefasciatus: 62.00% vs. 34.25%; Ae. albopictus: 50.75% vs 45.00%, An. sinensis: 49.75% vs 39.00%). Mosquitoes captured by the MK with heaters at 365 nm were significantly more than those captured by the MK without heaters at 365 nm. A trap with a 365 nm wavelength, heating element, and lure showed significantly better capture effectiveness than MK with a 365 nm wavelength, heating element, but without lure (Cx. quinquefasciatus: 67.00% vs. 29.75%, Ae. albopictus: 60.25% vs 36.25%, An. sinensis: 49.75% vs 39.75%). The coated photocatalyst trap with a 365 nm wavelength, heating element, and lure showed significantly better capture effectiveness than the trap without coating (Cx. quinquefasciatus: 54.25% vs. 42.50%; Ae. albopictus: 53.50% vs 44.00%, An. sinensis: 50.00% vs 41.25%). This trap demonstrated a significantly better capture advantage for Cx. quinquefasciatus and Ae. albopictus compared to the three commercial products. CONCLUSION: The developed mosquito trap with multiple attractant factors significantly enhanced the capture effectiveness of common mosquitoes. © 2024 Society of Chemical Industry. [ABSTRACT FROM AUTHOR]

Background: The efficacy of vector control tools depends on the behavior of the vector species. Many studies have sought to determine the feeding behavior of Anopheles mosquitoes in different settings of Ethiopia. We have performed a systematic review aimed to generate pooled evidence on the overall and species-specific blood meal sources of Anopheles mosquitoes in Ethiopia. Methods: A search for relevant articles was performed in two electronic databases (PubMed and Science Direct) and three search engines (Google Scholar, Research Gate and Google) between 11 March and 2 April 2024. Following the initial identification of articles, we used EndNote X8 software and removed duplicate articles and screened the remaining articles by careful reading of their titles and abstracts. The full text of articles that passed this screening phase was retrieved, read and evaluated against predetermined selection criteria. The final decision for inclusion in the systematic review was made after a methodological quality check using the JBI critical appraisal checklist. All relevant data were extracted from tables, figures and texts of the included articles using a premade template in Excel, and the data were analyzed using Stata version 14 software. Results: Of the 2431 studies identified, 27 met the inclusion criteria; all were published between 1997 and 2024. At 215 data points (frequency of tests of each Anopheles species by location and method of mosquito collections), 18,771 Anopheles mosquitoes belonging to 23 species or species complexes were tested for blood meal sources. The commonest sources of blood meals for Anopheles mosquitoes were bovine (36.0%, n = 6758) and human (29.4%, n = 5520). Among the tested anophelines, Anopheles (An.) arabiensis accounted for 67.9% (n = 12,741), followed by An. pharoensis, An. demeilloni and An. stephensi at 10.0%, 5.6% and 4.4%, respectively. Overall, there was no difference in the mean proportion of An. arabiensis detected with domestic animal blood (33.4%, 95% confidence interval [CI] 32.4–34.4%) and those detected with human blood (31.8%, 95% CI 30.9–32.8%). However, a greater proportion of the outdoor collected An. arabiensis were found to feed on bovines (47.9%, 95% CI 35.3–60.6) compared to humans (12.9%, 95% CI 0.8–24.9, P < 0.01). The foraging ratio (FR), which accounts for host availability, was greater for bovines (FR = 0.7) than for humans (FR = 0.2) for An. arabiensis, indicating preferential feeding on bovine hosts. This host preference was supported by the host preference index (human:bovine = 0.4). Anopheles pharoensis was detected with a slightly higher human blood index (53.5%, n = 1005) compared to bovine blood index (45.2%, n = 849). In contrast, An. demeilloni, An. coustani and An. marshalli were detected with a higher bovine blood index. Recently invaded urban malaria vector, An. stephensi was found with a higher ovine blood index. Conclusions: Bovine and human hosts are common sources of a blood meal for Anopheles mosquitoes. In terms of host availability, An. arabiensis showed preferential feeding on bovines/cattle. Targeting domestic animals, bovines and ovines with endectocides could supplement current vector control interventions. Study registration: The protocol of this study was registered on the International Prospective Register of Systematic Reviews, registration no. CRD42024515725. [ABSTRACT FROM AUTHOR]

The main vectors of Zika virus (ZIKV) and dengue virus (DENV) are Aedes aegypti and Ae. albopictus, with Ae. aegypti being more competent. However, the underlying mechanisms remain unclear. Here, we find Ae. albopictus shows comparable vector competence to ZIKV/DENV with Ae. aegypti by blood-feeding after antibiotic treatment or intrathoracic injection. This suggests that midgut microbiota can influence vector competence. Enterobacter hormaechei_B17 (Eh_B17) is isolated from field-collected Ae. albopictus and conferred resistance to ZIKV/DENV infection in Ae. aegypti after gut-transplantation. Sphingosine, a metabolite secreted by Eh_B17, effectively suppresses ZIKV infection in both Ae. aegypti and cell cultures by blocking viral entry during the fusion step, with an IC50 of approximately 10 μM. A field survey reveals that Eh_B17 preferentially colonizes Ae. albopictus compared to Ae. aegypti. And field Ae. albopictus positive for Eh_B17 are more resistant to ZIKV infection. These findings underscore the potential of gut symbiotic bacteria, such as Eh_B17, to modulate the arbovirus vector competence of Aedes mosquitoes. As a natural antiviral agent, Eh_B17 holds promise as a potential candidate for blocking ZIKV/DENV transmission. Here the authors show that arbovirus vector competence of Aedes mosquitoes is modulated by the midgut microbiota and identify the metabolite sphingosine from the gut symbiotic bacterium Enterobacter hormaechei_B17 (Eh_B17) as a factor mediating resistance to ZIKV and DENV infection. [ABSTRACT FROM AUTHOR]

Background: The 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. Methods: A 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. Results: The 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). Conclusions: The 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. [ABSTRACT FROM AUTHOR]

Mosquitoes transmit viruses that affect public health in tropical regions. Identifying factors that influence their fitness can help control mosquito-borne diseases. We investigated the impacts of predation risk effects, from a fish predator, and food availability on the oviposition behavior of Aedes aegypti. We hypothesize that predation risk and food availability interactively affect Ae. aegypti oviposition. Gravid Ae. aegypti females were offered oviposition sites with all possible combinations of predation risk (with vs. without predator cues) and food availability (low vs. high). We found a significant interaction between predation risk and food availability. The magnitude of oviposition and the probability of egg laying in high food availability treatments exceeded the values of low food availability treatments, but only in treatments without predation risk. Predation risk critically modulated the effects of food availability on the oviposition of Ae. aegypti. This study demonstrates the interplay between predation risk and food availability in controlling mosquito-borne diseases. [ABSTRACT FROM AUTHOR]

In recent decades, field and semi-field studies of malaria transmission have gathered geographic-specific information about mosquito ecology, behaviour and their sensitivity to interventions. Mathematical models of malaria transmission can incorporate such data to infer the likely impact of vector control interventions and hence guide malaria control strategies in various geographies. To facilitate this process and make model predictions of intervention impact available for different geographical regions, we developed AnophelesModel. AnophelesModel is an online, open-access R package that quantifies the impact of vector control interventions depending on mosquito species and location-specific characteristics. In addition, it includes a previously published, comprehensive, curated database of field entomological data from over 50 Anopheles species, field data on mosquito and human behaviour, and estimates of vector control effectiveness. Using the input data, the package parameterizes a discrete-time, state transition model of the mosquito oviposition cycle and infers species-specific impacts of various interventions on vectorial capacity. In addition, it offers formatted outputs ready to use in downstream analyses and by other models of malaria transmission for accurate representation of the vector-specific components. Using AnophelesModel, we show how the key implications for intervention impact change for various vectors and locations. The package facilitates quantitative comparisons of likely intervention impacts in different geographical settings varying in vector compositions, and can thus guide towards more robust and efficient malaria control recommendations. The AnophelesModel R package is available under a GPL-3.0 license at https://github.com/SwissTPH/AnophelesModel. [ABSTRACT FROM AUTHOR]

On August 27, 2024, this report was posted as an MMWR Early Release on the MMWR website (https://www.cdc.gov/mmwr). Beginning in late 2023, Oropouche virus was identified as the cause of large outbreaks in Amazon regions with known endemic transmission and in new areas in South America and the Caribbean. The virus is spread to humans by infected biting midges and some mosquito species. Although infection typically causes a self-limited febrile illness, reports of two deaths in patients with Oropouche virus infection and vertical transmission associated with adverse pregnancy outcomes have raised concerns about the threat of this virus to human health. In addition to approximately 8,000 locally acquired cases in the Americas, travel-associated Oropouche virus disease cases have recently been identified in European travelers returning from Cuba and Brazil. As of August 16, 2024, a total of 21 Oropouche virus disease cases were identified among U.S. travelers returning from Cuba. Most patients initially experienced fever, myalgia, and headache, often with other symptoms including arthralgia, diarrhea, nausea or vomiting, and rash. At least three patients had recurrent symptoms after the initial illness, a common characteristic of Oropouche virus disease. Clinicians and public health jurisdictions should be aware of the occurrence of Oropouche virus disease in U.S. travelers and request testing for suspected cases. Travelers should prevent insect bites when traveling, and pregnant persons should consider deferring travel to areas experiencing outbreaks of Oropouche virus disease. [ABSTRACT FROM AUTHOR]

Our knowledge of alphavirus genetic diversity is mainly based on viruses isolated from anthropophilic mosquito species, humans, and livestock during outbreaks. Studies on alphaviruses from sylvatic amplification cycles in sub-Saharan Africa have been conducted less often than from epizootic environments. To gain insight into alphavirus diversity in enzootic transmission cycles, we collected over 23,000 mosquitoes in lowland rainforest and savannah gallery forest in southwestern Uganda and tested them for alphavirus infections. We detected Sindbis virus (SINV) in a Culex Culex sp. mosquito and Middelburg virus (MIDV) in Eretmapodites intermedius and Mansonia africana. MIDV is a mosquito-borne alphavirus that causes febrile illness in sheep, goats, and horses and was previously not known to occur in Uganda. SINV, also a mosquito-borne alphavirus, causes mild infections in humans. Full genomes of SINV and MIDV were sequenced, showing a nucleotide identity of 99% to related strains. Both isolates replicated to high titres in a wide variety of vertebrate cells. Our data suggest endemic circulation of SINV and MIDV in Uganda. [ABSTRACT FROM AUTHOR]

Mosquito-borne diseases are a major global health threat. Traditional morphological or molecular methods for identifying mosquito species often require specialized expertise or expensive laboratory equipment. The use of convolutional neural networks (CNNs) to identify mosquito species based on images may offer a promising alternative, but their practical implementation often remains limited. This study explores the applicability of CNNs in classifying mosquito species. It compares the efficacy of body and wing depictions across three image collection methods: a smartphone, macro-lens attached to a smartphone and a professional stereomicroscope. The study included 796 specimens of four morphologically similar Aedes species, Aedes aegypti, Ae. albopictus, Ae. koreicus and Ae. japonicus japonicus. The findings of this study indicate that CNN models demonstrate superior performance in wing-based classification 87.6% (95% CI: 84.2–91.0) compared to body-based classification 78.9% (95% CI: 77.7–80.0). Nevertheless, there are notable limitations of CNNs as they perform reliably across multiple devices only when trained specifically on those devices, resulting in an average decline of mean accuracy by 14%, even with extensive image augmentation. Additionally, we also estimate the required training data volume for effective classification, noting a reduced requirement for wing-based classification compared to body-based methods. Our study underscores the viability of both body and wing classification methods for mosquito species identification while emphasizing the need to address practical constraints in developing accessible classification systems. [ABSTRACT FROM AUTHOR]

Background: C-type lectins (CTLs) are a large family of proteins with sugar-binding activity. CTLs contain an evolutionarily conserved C-type lectin domain (CTLD) that binds microbial carbohydrates in a calcium-dependent manner, thereby playing a key role in both microbial pathogenesis and innate immune responses. Aedes albopictus is an important vector for transmitting dengue virus (DENV) worldwide. Currently, the molecular characteristics and functions of CTLs in Ae. albopictus are largely unknown. Methods: Transcripts encoding CTL proteins in the Ae. albopictus genome assembly were analyzed via sequence blast. Phylogenetic analysis and molecular characterization were performed to identify the functional domains of the CTLs. Quantitative analysis was performed to determine the gene expression features of CTLs during mosquito development and in different tissues of female adults after blood feeding. In addition, the functional role of CTLs in response to DENV infection was investigated in Ae. albopictus mosquito cells. Results: We identified 39 transcripts encoding CTL proteins in the Ae. albopictus transcriptome. Aedes albopictus CTLs are classified into three groups based on the number of CTLDs and the domain architecture. These included 29 CTL-Ss (single-CTLDs), 1 immulectins (dual-CTLD) and 9 CTL-Xs (CTLDs with other domains). Phylogenetic analysis and structural modeling indicated that CTLs in Ae. albopictus are highly conserved with the homologous CTLs in Aedes aegypti. The expression profile assay revealed differential expression patterns of CTLs in both developmental stages and in adult female tissues. Knockdown and overexpression of three CTLs (CTL-S12, S17 and S19) confirmed that they can promote dengue virus infection in Ae. albopictus cells. Conclusions: The CTL genes in Ae. albopictus mosquito and other mosquito species are evolutionarily conserved and exhibit different developmental and tissue expression features. The functional assay indicated that three CTLs in Ae. albopictus mosquitoes are involved in promoting dengue virus infection. Our study revealed that CTLs play important roles in both the physiological processes and viral infection in mosquito vectors. [ABSTRACT FROM AUTHOR]

The mosquito microbiome significantly influences vector competence, including in Aedes albopictus, a globally invasive vector. Describing the microbiome and Wolbachia strains of Ae. albopictus from different regions can guide area-specific control strategies. Mosquito samples from Spain and São Tomé were analyzed using 16S rRNA gene sequencing and metagenomic sequencing. Wolbachia infection patterns were observed by sex and population. Female mosquitoes were blood-fed, a factor considered in analyzing their microbiota. Results revealed a dominance of dual Wolbachia infections, strains A and B, in the microbiome of both populations of Ae. albopictus, especially among females. Both populations shared a core microbiome, although 5 and 9 other genera were only present in Spain and São Tomé populations, respectively. Genera like Pelomonas and Nevskia were identified for the first time in Aedes mosquitoes. This study is the first to describe the Ae. albopictus bacteriome in Spain and São Tomé, offering insights for the development of targeted mosquito control strategies. Understanding the specific microbiome composition can help in designing more effective interventions, such as microbiome manipulation and Wolbachia-based approaches, to reduce vector competence and transmission potential of these mosquitoes. [ABSTRACT FROM AUTHOR]

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The monitoring of mosquitoes is of great importance due to their vector competence for a variety of pathogens, which have the potential to imperil human and animal health. Until now mosquito occurrence data is mainly obtained with conventional monitoring methods including active and passive approaches, which can be time- and cost-consuming. New monitoring methods based on environmental DNA (eDNA) could serve as a fast and robust complementary detection system for mosquitoes. In this pilot study already existing marker systems targeting the three invasive mosquito species Aedes (Ae.) albopictus, Ae. japonicus and Ae. koreicus were used to detect these species from water samples via microfluidic array technology. We compared the performance of the high-throughput real-time PCR (HT-qPCR) system Biomark HD with real-time PCR (qPCR) and also tested the effect of different filter media (Sterivex® 0.45 µm, Nylon 0.22 µm, PES 1.2 µm) on eDNA detectability. By using a universal qPCR protocol and only 6-FAM-MGB probes we successfully transferred these marker systems on the HT-qPCR platform. All tested marker systems detected the target species at most sites, where their presence was previously confirmed. Filter media properties, the final filtration volume and observed qPCR inhibition did not affect measured Ct values via qPCR or HT-qPCR. The Ct values obtained from HT-qPCR were significantly lower as Ct values measured by qPCR due to the previous preamplification step, still these values were highly correlated. Observed incongruities in eDNA detection probability, as manifested by non-reproducible results and false positive detections, could be the result of methodological aspects, such as sensitivity and specificity issues of the used assays, or ecological factors such as varying eDNA release patterns. In this study, we show the suitability of eDNA-based detection of mosquito species from water samples using a microfluidic HT-qPCR platform. HT-qPCR platforms such as Biomark HD allow for massive upscaling of tested species-specific assays and sampling sites with low time- and cost-effort, thus this methodology could serve as basis for large-scale mosquito monitoring attempts. The main goal in the future is to develop a robust (semi)-quantitative microfluidic-based eDNA mosquito chip targeting all haematophagous culicid species occurring in Western Europe. This chip would enable large-scale eDNA-based screenings to assess mosquito diversity, to monitor species with confirmed or suspected vector competence, to assess the invasion progress of invasive mosquito species and could be used in pathogen surveillance, when disease agents are incorporated. [ABSTRACT FROM AUTHOR]

Abundance and seasonal composition of adult indoor resting Culicine mosquitoes in Dutse were studied between April to September 2021. Culex pipiens complex and Aedes species has been proven as the potential vectors of public health importance. However, their abundance and seasonal composition has not been adequately studied in Dutse, Northwest Nigeria to document their vectorial profile. This study was designed to fill the gap. Adult indoor resting mosquitoes were collected using Pyrethrum Spray catch (PSC) from April - September 2021 and were morphologically and molecularly identified using keys and polymerase chain reaction (PCR). Data generated were analysed using Statistical Package for Social Sciences (SPSS). 2656 mosquitoes, comprising Culex quinquefasciatus (2507) and Aedes species (149) were collected. Culex quinquefasciatus was the most abundant species (94.4%) collected while the least were Aedes species (5.6%). The results revealed that in Jidawa, more Culex quinquefasciatus (80) were collected in August while the least were collected in June (25). For Aedes mosquitoes, higher collection (75) was realised in April while July recorded zero collection. The monthly variation of the mosquitoes showed the peak in the month of April (690) and May (300) in Jidawa when compared to Kargo (430) and (100) respectively. In the dry season, more mosquitoes were collected In Jidawa while in Kargo more were collected during the wet seasons. The result of the molecular identification of the Culex pipiens complex reveals Culex quinquefasciatus as the only sibling species in both Jigawa and Kargo. The occurrence of Culex quinquefasciatus and Aedes species is an indication of the signal against lymphatic filariasis, dengue fever, arbovirus and yellow fever in the area. Therefore, there is need for vector control programmes and public orientation on the drivers of mosquito abundance in the study area, taking into cognisance the seasonality variation of the mosquitoes. [ABSTRACT FROM AUTHOR]

Understanding the biting behaviour of vectors is essential for determining their vectorial potential and, hence, their role in disease transmission. The study was a two-year entomological survey of malaria vectors in three agrarian communities of Kano state designated as large irrigation (LIC), urban irrigation (UIC) and nonirrigation (NIC) communities. Human-baited Centre for Disease Control (CDC) light traps collected outdoor and indoor biting adult mosquitoes overnight. The collected female Anopheles species were morphologically identified to the species level using standard keys. A total of six hundred and thirty-seven (637) female Anopheles species were trapped of which 322 (50.55%) were collected indoors and 315 (49.45%) were collected outdoors. LIC and NIC had more Anopheles mosquitoes collected outdoors (51.08% and 66.04% respectively) than indoors (48.92% and 33.96% respectively) while UIC had more Anopheles mosquitoes collected indoors (67.77%) than outdoors (32,23%). Mosquito biting activity was higher in the wet season than in the dry season but did not differ significantly indoors (F-value = 3.310, p = 0.84) and outdoors (F = 0.651, p = 0.532). Statistical differences were recorded in the mean outdoor Anopheles abundance (F = 4.51 and p = 0.044) across the communities. Single, bimodal, and multiple biting peaks were observed in the study. Early biting peaks (1800-1900 hours) were observed outdoors (wet season) in NIC and indoors in UIC (dry season). The early outdoor biting activities of Anopheles species established in this study suggest the possibility of continued malarial transmission despite household use of bed nets. This implies that this control measure might be insufficient for malaria control in the region, and thus integrated methods for controlling malaria vectors should be considered. [ABSTRACT FROM AUTHOR]