Pseudomonas protegens Affects Mosquito Survival and Development.

This study investigated the pathogenic potential of Pseudomonas protegens on mosquito larvae of the two species Culex pipiens and Aedes albopictus, representing major threats for disease transmission in the Mediterranean area and worldwide. The bacterium achieved to kill over 90% of the mosquito larvae within 72 h after exposition to a bacterial concentration of 100 million CFU/ml. These lethal effects were concentration dependent and a significantly higher susceptibility was associated with younger larvae of both mosquito species. Significant slowdown of immature (larval and pupal) development and decrease in adult emergence rate after treatment with sub-lethal doses of the bacterium were also detected. This study reports for the first time the insecticidal activity of a root-associated biocontrol bacterium against aquatic mosquito larvae.

Experiências em divulgação científica e sensibilização da população: importância do controle mecânico do vetor Aedes aegypti / Experiences in scientific outreach and public awareness: the importance of mechanical control of the Aedes aegypti vector / Experiencias en divulgación científica y sensibilización de la población: importancia del control mecánico del vector Aedes aegypti

Aedes aegypti é o principal vetor dos agentes etiológicos de dengue, zika e chikungunya, doenças para as quais não existem vacinas totalmente eficazes. Alternativas de controle visando mitigar essas arboviroses são primordiais. Entre essas, o controle mecânico aborda práticas de eliminação e/ou limpeza de criadouros do vetor. Neste relato, apresentamos e avaliamos criticamente ações realizadas pelo grupo, ocorridas entre 2016 e 2019, nas quais divulgamos informação científica clara através do diálogo com a população. Os métodos utilizados foram: 1) palestras em escolas (público infantojuvenil) utilizando slides, fotos e vídeos; 2) oficinas (público misto), estande com material in vivo do ciclo de vida do Aedes, jogos e desenhos. Analisamos dez palestras em escolas do ensino fundamental e médio e vinte oficinas realizadas em diferentes regiões do Brasil. Concluímos que tais ações e suas análises críticas devem ser realizadas continuamente para que sejam bem-sucedidas

Aedes aegypti is the main vector of dengue, zika, and chikungunya etiological agents, diseases for which no effective vaccines are available. Control alternatives aimed at mitigating these arboviruses are essential. Among such, mechanical control addresses practices of elimination and/or cleaning of vector breeding sites. Here, we presented and critically evaluated actions carried out by ourselves. These actions took place between 2016 and 2019, where we disseminated clear scientific information through dialogue with the population. The following methods were employed: 1) lectures in schools (children and youth audiences) using slides, photos, and videos; 2) workshops (mixed audience), stand with in vivo material from the Aedeslife cycle, games, and drawings. Ten lectures in elementary and high schools and twenty workshops held in different regions of Brazil were analyzed. It was concluded that such actions and their critical analyzes must be carried out continuously to be successful

Aedes aegypti es el principal vector de los agentes etiológicos del dengue, zika y chikungunya, enfermedades para las que no existen vacunas totalmente eficaces. Las alternativas de control para mitigar estas arbovirosis son fundamentales. El control mecánico, representa una de estas alternativas, aborda prácticas de eliminación y/o limpieza de criaderos del vector. En este informe presentamos y evaluamos de manera crítica las acciones realizadas por el grupo entre los años 2016 y 2019. Presentamos información científica clara a través del diálogo con la población mediante los siguientes métodos: 1) conferencias en escuelas (público infantil) utilizando diapositivas, fotos y videos; 2) Talleres (público mixto), stand con material in vivo del ciclo de vida del Aedes, juegos y dibujos. Analizamos diez conferencias en escuelas (primarias y secundarias) y veinte talleres realizados en diferentes regiones de Brasil. Concluimos que tales acciones y el análisis crítico de las mismas deben llevarse a cabo de manera continua para que resulten exitosas.

Occurrence of the Sylvatic Yellow Fever Vector Mosquito Haemagogus leucocelaenus (Diptera: Culicidae) in an Atlantic Forest Fragment of the Touristic State of Rio de Janeiro, Brazil.

The yellow fever virus is estimated to cause 30,000 deaths each year worldwide, with the majority of cases and deaths occurring in Africa. The virus is also endemic to Central and South America, including northern and western Brazil. The sylvatic cycle of the virus is related to wild and rural areas, with nonhuman primates as the primary host and wild mosquitoes, specifically from the genera Haemagogus, as vectors. The diversity of the mosquito community plays a significant role in the increase of pathogen transmission to humans. In the present study, we detected fluctuation in populations of vector mosquitoes using ovitraps for Culicidae egg collection. The study area is a forest fragment of the Atlantic Forest, one of the most threatened biomes in Brazil. This biome has been suffering significant deforestation due to anthropic activity. Worryingly, the proximity of human populations to forest environments increases the risk of spreading disease from forest fragments to urban areas. Our findings showed that the highest egg abundance occurred in December 2019, with a significant difference (p = 0.005) between rainy and dry seasons. Most eggs were collected during the rainy period. Subsequent quantification of specimens from epidemiologically relevant species hatched from field-collected eggs resulted in 1,131 (86%) Haemagogus leucocelaenus (Dyar & Shannon, 1924), 111 (8%) Aedes terrens (Walker, 1856), 47 (4%) Aedes albopictus (Skuse, 1894), and 21 (2%) Haemagogus janthinomys (Dyar, 1921). Finally, we assessed the behavior of different vector species performing oviposition on the same breeding site. The highest correlation coefficient was observed between Ae. albopictus and Ae. terrens (rho = 0.52) concerning other Culicidae species. Therefore, we believe that Culicidae population surveillance is crucial for disease monitoring since the increase in specimens of a number of vector species influences the emergence of yellow fever cases in nonhuman primates and human populations.

Incidência de casos confirmados de dengue no Estado do Paraná Brasil nos anos de 2016 a 2021 / Incidence of confirmed dengue cases in the State of Paraná Brazil in the years of 2016 to 2021 / Incidencia de casos confirmados de dengue en el Estado de Paraná Brasil en los años 2016 a 2021

A dengue é uma doença dolorosa e debilitante transmitida por insetos da espécie Aedes aegypti. Ela é definida como uma doença viral que, nos últimos anos, se espalhou vertiginosamente por todas as regiões tropicais e subtropicais do planeta. Este estudo teve como objetivo identificar e discutir o número e a taxa de incidência de casos de dengue no estado do Paraná utilizando-se dos boletins emitidos por semana epidemiológica nos anos de 2016 a 2021, considerando a sazonalidade da doença. Também se objetivou debater a incidência por macrorregional, as possíveis causas de períodos epidêmicos e ações de combate vetorial para redução dos casos da patologia. Foram utilizados como fonte de informações o banco de dados da Dengue/SVS/SESA, por meio de informes técnicos, disponibilizados pelo portal online de Boletins da Dengue Paraná da Secretaria de Estado de Saúde do Paraná. Conclui-se que o ano epidemiológico de 2019/2020 foi o de maior incidência e os anos epidemiológicos 2016/2017 e 2017/2018 apresentaram os menores casos durante todo período analisado. Dessa forma, a vigilância epidemiológica é muito importante para avaliação espacial da distribuição de casos para execução de ações estratégicas para redução da infestação do vetor. As políticas públicas e a disponibilização de inseticidas para aplicação também são essenciais para o combate da Dengue.

Dengue is a painful and debilitating disease transmitted by insects of the Aedes aegypti species. It is defined as a viral disease that, in recent years, has spread vertiginously throughout the tropical and subtropical regions of the planet. This study aimed to identify and discuss the number and incidence rate of dengue cases in the state of Paraná using the bulletins issued by epidemiological week in the years 2016 to 2021, considering the seasonality of the disease. The aim was also to discuss the incidence per macro-region, the possible causes of epidemic periods, and vectorial combat actions to reduce the cases of the pathology. The Dengue/SVS/SESA database was used as a source of information, through technical reports, made available by the online portal of Dengue Paraná Bulletins of the Paraná State Health Department. It is concluded that the epidemiological year 2019/2020 was the one with the highest incidence and the epidemiological years 2016/2017 and 2017/2018 had the lowest cases during the entire period analyzed. Thus, epidemiological surveillance is very important for the spatial assessment of the distribution of cases to carry out strategic actions to reduce vector infestation. Public policies and the availability of insecticides for application are also essential to combat Dengue.

El dengue es una enfermedad dolorosa y debilitante transmitida por insectos de la especie Aedes aegypti. Se define como una enfermedad viral que, en los últimos años, se ha extendido vertiginosamente por las regiones tropicales y subtropicales del planeta. Este estudio tuvo como objetivo identificar y discutir el número y la tasa de incidencia de los casos de dengue en el estado de Paraná utilizando los boletines emitidos por la semana epidemiológica en los años 2016 a 2021, considerando la estacionalidad de la enfermedad. También se pretendía discutir la incidencia por macrorregiones, las posibles causas de los periodos epidémicos y las acciones de control de vectores para la reducción de los casos de la enfermedad. Se utilizó como fuente de información la base de datos de Dengue/SVS/SESA, por medio de informes técnicos, puestos a disposición por el portal online de Boletines de Dengue Paraná de la Secretaría de Salud del Estado de Paraná. Se concluye que el año epidemiológico 2019/2020 fue el de mayor incidencia y los años epidemiológicos 2016/2017 y 2017/2018 presentaron los menores casos durante todo el periodo analizado. Por lo tanto, la vigilancia epidemiológica es muy importante para la evaluación espacial de la distribución de los casos para la implementación de acciones estratégicas para reducir la infestación del vector. Las políticas públicas y la disponibilidad de insecticidas para su aplicación también son esenciales para combatir el dengue.

Essential functions of mosquito ecdysone importers in development and reproduction.

The primary insect steroid hormone ecdysone requires a membrane transporter to enter its target cells. Although an organic anion-transporting polypeptide (OATP) named Ecdysone Importer (EcI) serves this role in the fruit fly Drosophila melanogaster and most likely in other arthropod species, this highly conserved transporter is apparently missing in mosquitoes. Here we report three additional OATPs that facilitate cellular incorporation of ecdysone in Drosophila and the yellow fever mosquito Aedes aegypti. These additional ecdysone importers (EcI-2, -3, and -4) are dispensable for development and reproduction in Drosophila, consistent with the predominant role of EcI. In contrast, in Aedes, EcI-2 is indispensable for ecdysone-mediated development, whereas EcI-4 is critical for vitellogenesis induced by ecdysone in adult females. Altogether, our results indicate unique and essential functions of these additional ecdysone importers in mosquito development and reproduction, making them attractive molecular targets for species- and stage-specific control of ecdysone signaling in mosquitoes.

Ecological and environmental factors affecting transmission of sylvatic yellow fever in the 2017-2019 outbreak in the Atlantic Forest, Brazil.

BACKGROUND: Yellow fever virus (YFV) is an arbovirus that, despite the existence of a safe and effective vaccine, continues to cause outbreaks of varying dimensions in the Americas and Africa. Between 2017 and 2019, Brazil registered un unprecedented sylvatic YFV outbreak whose severity was the result of its spread into zones of the Atlantic Forest with no signals of viral circulation for nearly 80 years. METHODS: To investigate the influence of climatic, environmental, and ecological factors governing the dispersion and force of infection of YFV in a naïve area such as the landscape mosaic of Rio de Janeiro (RJ), we combined the analyses of a large set of data including entomological sampling performed before and during the 2017-2019 outbreak, with the geolocation of human and nonhuman primates (NHP) and mosquito infections. RESULTS: A greater abundance of Haemagogus mosquitoes combined with lower richness and diversity of mosquito fauna increased the probability of finding a YFV-infected mosquito. Furthermore, the analysis of functional traits showed that certain functional groups, composed mainly of Aedini mosquitoes which includes Aedes and Haemagogus mosquitoes, are also more representative in areas where infected mosquitoes were found. Human and NHP infections were more common in two types of landscapes: large and continuous forest, capable of harboring many YFV hosts, and patches of small forest fragments, where environmental imbalance can lead to a greater density of the primary vectors and high human exposure. In both, we show that most human infections (~ 62%) occurred within an 11-km radius of the finding of an infected NHP, which is in line with the flight range of the primary vectors. CONCLUSIONS: Together, our data suggest that entomological data and landscape composition analyses may help to predict areas permissive to yellow fever outbreaks, allowing protective measures to be taken to avoid human cases.

A comparative study of dengue virus vectors in major parks and adjacent residential areas in Ho Chi Minh City, Vietnam.

The primary dengue virus vectors, Aedes aegypti and Aedes albopictus, are primarily daytime biting mosquitoes. The risk of infection is suspected to be considerable in urban parks due to visitor traffic. Despite the importance of vector control for reducing dengue transmission, little information is available on vector populations in urban parks. The present study characterized mosquito habitats and estimated vector densities in the major urban parks in Ho Chi Minh City, Vietnam and compared them with those in adjacent residential areas. The prevalences of habitats where Aedes larvae were found were 43% and 9% for the parks and residential areas, respectively. The difference was statistically significant (prevalence ratio [PR]: 5.00, 95% CI: 3.85-6.49). The prevalences of positive larval habitats were significantly greater in the parks for both species than the residential areas (PR: 1.52, 95% CI: 1.04-2.22 for A. aegypti, PR: 10.10, 95% CI: 7.23-14.12 for A. albopictus). Larvae of both species were positively associated with discarded containers and planters. Aedes albopictus larvae were negatively associated with indoor habitats, but positively associated with vegetation shade. The adult density of A. aegypti was significantly less in the parks compared with the residential areas (rate ratio [RR]; 0.09, 95% CI: 0.05-0.16), while the density of A. albopictus was significantly higher in the parks (RR: 9.99, 95% CI: 6.85-14.59). When the species were combined, the density was significantly higher in the parks (RR: 2.50, 95% CI: 1.92-3.25). The urban parks provide suitable environment for Aedes mosquitoes, and A. albopictus in particular. Virus vectors are abundant in the urban parks, and the current vector control programs need to have greater consideration of urban parks.

Competition and resource depletion shape the thermal response of population fitness in Aedes aegypti.

Mathematical models that incorporate the temperature dependence of lab-measured life history traits are increasingly being used to predict how climatic warming will affect ectotherms, including disease vectors and other arthropods. These temperature-trait relationships are typically measured under laboratory conditions that ignore how conspecific competition in depleting resource environments-a commonly occurring scenario in nature-regulates natural populations. Here, we used laboratory experiments on the mosquito Aedes aegypti, combined with a stage-structured population model, to investigate this issue. We find that intensified larval competition in ecologically-realistic depleting resource environments can significantly diminish the vector's maximal population-level fitness across the entire temperature range, cause a ~6 °C decrease in the optimal temperature for fitness, and contract its thermal niche width by ~10 °C. Our results provide evidence for the importance of considering intra-specific competition under depleting resources when predicting how arthropod populations will respond to climatic warming.

Exposure to copper sulfate impairs survival, post-embryonic midgut development and reproduction in Aedes aegypti.

Aedes aegypti is a vector of several global human viruses responsible for high human morbidity and mortality. The method to prevent the transmission of vector-borne viruses is mainly based on the control of the insect vector using insecticides. Among these chemicals, copper sulfate is a compound widely used in agriculture with the potential to be used as an alternative to control these insects. This study evaluated the effects of the exposure of A. aegypti larvae to copper sulfate on survival, midgut morphology, blood-feeding and fecundity. The exposure to CuSO4 decreased the survival of A. aegypti during the immature phase. Adults obtained from exposed larvae had their lifespan decreased at all tested concentrations. The exposure to CuSO4 impaired the development in the transition from larvae to pupae and from pupae to adult. The number of eggs laid by females developed from larvae treated with CuSO4 was significantly lower than in control. In addition, the egg hatching rates were also negatively affected. The midguts of treated larvae and pupae showed epithelial disorganization. The number of cleaved caspase-3 cells increased in the midgut of exposed pupae compared to control. Moreover, there was a reduction in proliferating cells in treated larvae and pupae compared to the control. In conclusion, the results reveal that CuSO4 exposure has insecticidal activity against A. aegypti, which may be related to the impairment of the midgut metamorphosis and reduced proliferation of stem cells, with the consequent impairment of female mosquito fertility and fecundity.

Statewide Efficacy Assessment of Insect Growth Regulators Against Aedes albopictus (Diptera: Culicidae) in Sabah, Malaysia: An Alternative Control Strategy?

The efficacy of three groups of insect growth regulators, namely juvenile hormone mimics (methoprene and pyriproxyfen), chitin synthesis inhibitors (diflubenzuron and novaluron), and molting disruptor (cyromazine) was evaluated for the first time, against Aedes albopictus Skuse (Diptera: Culicidae) larvae from 14 districts in Sabah, Malaysia. The results showed that all field populations of Ae. albopictus were susceptible towards methoprene, pyriproxyfen, diflubenzuron, novaluron, and cyromazine, with resistance ratio values ranging from 0.50-0.90, 0.60-1.00, 0.67-1.17, 0.71-1.29, and 0.74-1.07, respectively. Overall, the efficacy assessment of insect growth regulators in this study showed promising outcomes and they could be further explored as an alternative to conventional insecticides.

Impact of Cymbopogon flexuosus (Poaceae) essential oil and primary components on the eclosion and larval development of Aedes aegypti.

The current study describes the effects of sub-lethal concentrations and constituent compounds (citral and geranyl acetate) of Cymbopogon flexuosus essential oil (EO) on the development of Aedes aegypti. We treated eggs with 6, 18, or 30 mg L-1 and larvae with 3 or 6 mg L-1 of EO and its major compounds (citral and geranyl acetate). Citral and geranyl acetate were evaluated at 18, 30, and 42 mg L-1 and compared with commercial growth inhibitors (diflubenzuron and methoprene). We measured larval head diameter, siphon length, and larval length. Finally, we examined concentrations of molt hormone (MH) and juvenile hormone III (JH III) using high-performance liquid chromatography coupled to mass spectrometry. All geranyl acetate concentrations decreased egg hatching, while EO altered molting among larval instars and between larvae and pupae, with an increase in the larval length (3 mg L-1: 6 ± 0.0 mm; 6 mg L-1: 6 ± 0.7 mm) and head width (3 mg L-1: 0.8 ± 0 mm; 6 mg L-1: 0.8 ± 0.0 mm) compared with the control group. We did not detect chromatographic signals of MH and JH III in larvae treated with C. flexuosus EO or their major compounds. The sub-lethal concentrations C. flexuosus EO caused a similar effect to diflubenzuron, namely decreased hormone concentrations, an extended larval period, and death.

Aedes albopictus life table: environment, food, and age dependence survivorship and reproduction in a tropical area.

BACKGROUND: Environmental conditions affect the biology of mosquito vectors. Aedes albopictus is a major vector of many important diseases including dengue, Zika, and chikungunya in China. Understanding the development, fecundity, and survivorship of Ae. albopictus mosquitoes in different environmental conditions is beneficial for the implementation of effective vector control measures. METHODS: Aedes albopictus larval and adult life-table experiments were conducted under natural conditions in indoor, half-shaded, and fully shaded settings, simulating the three major habitat types in Hainan Province, a tropical island in the South China Sea. Temperature, humidity, and light intensity were recorded daily. Larval rearing used habitat water and tap water, with and without additional artificial food. Development time, survivorship, pupation rate, and adult emergence rates were monitored. Adult mosquito survivorship and fecundity were monitored daily and reproductive rates were determined, and age-dependent survivorship and reproduction were analyzed. RESULTS: The pupation time and male and female emergence times were significantly shorter in indoor conditions than in shaded and half-shaded conditions for both tap water with added food and habitat water with added food groups. For habitat water with added food, the shaded environment had the lowest pupation rate among the settings. For tap water with added food group, the shaded environment had the lowest pupation rate. The mean survival time of females was 27.3 ± 0.8 days in the indoor condition, which was significantly longer than that in the half-shaded (18.4 ± 0.6 days) and shaded (13.8 ± 1.2 days) conditions. Adult mortality was age-dependent, and the rate of change in mortality with age was not significantly different among different environmental conditions. The mean net replacement rate (R0) of female mosquitoes showed no significant difference among the three conditions, whereas the per capita intrinsic growth rate (r) in the shaded condition was 42.0% and 20.4% higher than that in the indoor and half-shaded conditions, respectively. Female daily egg mass was also age-dependent in all the settings, decaying exponentially with age. CONCLUSIONS: Our results imply that half-shaded conditions are likely the best natural condition for adult emergence and female reproduction, and food supply is crucial for larval development and pupation. The results provide new avenues for integrated mosquito management in indoor and outdoor areas, especially in half-shaded areas.

High Temperature Cycles Result in Maternal Transmission and Dengue Infection Differences Between Wolbachia Strains in Aedes aegypti.

Environmental factors play a crucial role in the population dynamics of arthropod endosymbionts, and therefore in the deployment of Wolbachia symbionts for the control of dengue arboviruses. The potential of Wolbachia to invade, persist, and block virus transmission depends in part on its intracellular density. Several recent studies have highlighted the importance of larval rearing temperature in modulating Wolbachia densities in adults, suggesting that elevated temperatures can severely impact some strains, while having little effect on others. The effect of a replicated tropical heat cycle on Wolbachia density and levels of virus blocking was assessed using Aedes aegypti lines carrying strains wMel and wAlbB, two Wolbachia strains currently used for dengue control. Impacts on intracellular density, maternal transmission fidelity, and dengue inhibition capacity were observed for wMel. In contrast, wAlbB-carrying Ae. aegypti maintained a relatively constant intracellular density at high temperatures and conserved its capacity to inhibit dengue. Following larval heat treatment, wMel showed a degree of density recovery in aging adults, although this was compromised by elevated air temperatures. IMPORTANCE In the past decades, dengue incidence has dramatically increased all over the world. An emerging dengue control strategy utilizes Aedes aegypti mosquitoes artificially transinfected with the bacterial symbiont Wolbachia, with the ultimate aim of replacing wild mosquito populations. However, the rearing temperature of mosquito larvae is known to impact on some Wolbachia strains. In this study, we compared the effects of a temperature cycle mimicking natural breeding sites in tropical climates on two Wolbachia strains, currently used for open field trials. When choosing the Wolbachia strain to be used in a dengue control program it is important to consider the effects of environmental temperatures on invasiveness and virus inhibition. These results underline the significance of understanding the impact of environmental factors on released mosquitoes, in order to ensure the most efficient strategy for dengue control.

Urbanization favors the proliferation of Aedes aegypti and Culex quinquefasciatus in urban areas of Miami-Dade County, Florida.

Urbanization processes are increasing globally. Anthropogenic alterations in the environment have profound effects on biodiversity. Decreased biodiversity due to biotic homogenization processes as a consequence of urbanization often result in increased levels of mosquito vector species and vector-borne pathogen transmission. Understanding how anthropogenic alterations in the environment will affect the abundance, richness, and composition of vector mosquito species is crucial for the implementation of effective and targeted mosquito control strategies. We hypothesized that anthropogenic alterations in the environment are responsible for increasing the abundance of mosquito species that are adapted to urban environments such as Aedes aegypti and Culex quinquefasciatus. Therefore, our objective was to survey mosquito relative abundance, richness, and community composition in Miami-Dade County, Florida, in areas with different levels of urbanization. We selected 24 areas, 16 remote areas comprised of natural and rural areas, and 8 urban areas comprised of residential and touristic areas in Miami-Dade County, Florida. Mosquitoes were collected weekly in each area for 24 h for 5 consecutive weeks from August to October 2020 using BG-Sentinel traps baited with dry ice. A total of 36,645 mosquitoes were collected, from which 34,048 were collected in the remote areas and 2,597 in the urban areas. Our results show a clear and well-defined pattern of abundance, richness, and community composition according to anthropogenic modifications in land use and land cover. The more urbanized a given area the fewer species were found and those were primary vectors of arboviruses, Ae. aegypti and Cx. quinquefasciatus.

The invasive Korean bush mosquito Aedes koreicus (Diptera: Culicidae) in Germany as of 2020.

BACKGROUND: The Korean bush mosquito Aedes koreicus was recently reported to have established a population in western Germany (Wiesbaden) in 2016. The species is difficult to distinguish morphologically from its close relative, the invasive Japanese bush mosquito Ae. japonicus, which is already widely distributed in many parts of Germany, including the area colonised by Ae. koreicus. Genetic confirmation of morphologically identified "Ae. japonicus" collection material, however, had only been done exceptionally before the German Ae. koreicus population became known. METHODS: Dried archived "Ae. japonicus" specimens both from the municipality of Wiesbaden and from deliberately and randomly selected distribution sites all over Germany were re-examined morphologically and genetically for admixture by Ae. koreicus. Moreover, cemeteries in the greater Wiesbaden area were sampled in 2019 and 2020 to check for Ae. koreicus spread. Korean and Japanese bush mosquitoes submitted to the German citizen science mosquito monitoring scheme "Mueckenatlas" in 2019 and 2020 were also subjected to particularly thorough species identification. The ND4 DNA sequences generated in this study in the context of species identification were phylogenetically compared to respective GenBank entries of Ae. koreicus. As a by-product, several genetic markers were evaluated for their suitability to identify Ae. koreicus. RESULTS: Aedes koreicus specimens could be identified in mosquito collection material and submissions from Wiesbaden from 2015 onwards, suggesting establishment to have happened in the same year as Ae. japonicus establishment. Detections of Ae. koreicus from 2019 and 2020 in Wiesbaden indicate a negligible enlargement of the populated area as described for 2018. Two Ae. koreicus specimens were also submitted from the city of Munich, southern Germany, in 2019 but further specimens could not be identified during immediate local inspections. Comparison of ND4 sequences generated in this and other studies demonstrate a high degree of homology, suggesting that this DNA region is not informative enough for clarification of origins and relationships of Ae. koreicus populations. For genetic identification of Ae. koreicus, PCR primers used for classical CO1 barcoding were found to lead to mismatches and produce no or incorrect amplicons. Alternative CO1 primers or a validated ND4 marker should be used instead. CONCLUSIONS: Aedes koreicus is probably introduced into Germany every now and then but rarely succeeds in becoming established. As with most European populations, the German population is characterised by a limited expansion tendency. Since Ae. koreicus is a potential vector, however, Asian bush mosquitoes found at new places should be examined quite carefully and known distribution areas of Ae. japonicus regularly checked for the presence of Ae. koreicus.

The first record of the invasive mosquito species Aedes albopictus in Chisinӑu, Republic of Moldova, 2020.

BACKGROUND: In Europe, Aedes albopictus is an important vector of chikungunya virus and Dirofilaria nematodes and has been involved in local autochthonous circulation of dengue and Zika viruses. Due to the ongoing spread, targeted field surveillance at potential points of entry of invasive Aedes mosquitoes was initiated by the Republic of Moldova in 2020 as part of the transboundary "Invasive Aedes Mosquitoes COST-Action project." METHODS: In 2020, ovitraps were positioned at each of three locations: the border crossing to Romania in Leuseni (Hancesti region), Chisinӑu International Airport and Chisinӑu Botanical Garden. RESULTS: A total of 188 Aedes spp. eggs were collected at the Chisinӑu International Airport between August and September 2020. Twenty-three adults reared in the laboratory were identified morphologically as Ae. albopictus (Skuse, 1895), and 12 selected specimens were confirmed by molecular barcoding of the cytochrome oxidase subunit I gene region. In addition, one adult Ae. albopictus female at the same site was caught with a manual aspirator. CONCLUSIONS: This is the first documented report of Ae. albopictus in the Republic of Moldova. The presence of immature and adult stages indicates the local reproduction of the species in the country. Therefore, it is crucial to extend and strengthen surveillance of the invasive Aedes mosquitoes to prevent Ae. albopictus and other exotic mosquito species from becoming established in the Republic of Moldova.

Modeling the effects of Aedes aegypti's larval environment on adult body mass at emergence.

Mosquitoes vector harmful pathogens that infect millions of people every year, and developing approaches to effectively control mosquitoes is a topic of great interest. However, the success of many control measures is highly dependent upon ecological, physiological, and life history traits of mosquito species. The behavior of mosquitoes and their potential to vector pathogens can also be impacted by these traits. One trait of interest is mosquito body mass, which depends upon many factors associated with the environment in which juvenile mosquitoes develop. Our experiments examined the impact of larval density on the body mass of Aedes aegypti mosquitoes, which are important vectors of dengue, Zika, yellow fever, and other pathogens. To investigate the interactions between the larval environment and mosquito body mass, we built a discrete time mathematical model that incorporates body mass, larval density, and food availability and fit the model to our experimental data. We considered three categories of model complexity informed by data, and selected the best model within each category using Akaike's Information Criterion. We found that the larval environment is an important determinant of the body mass of mosquitoes upon emergence. Furthermore, we found that larval density has greater impact on body mass of adults at emergence than on development time, and that inclusion of density dependence in the survival of female aquatic stages in models is important. We discuss the implications of our results for the control of Aedes mosquitoes and on their potential to spread disease.

Starvation at the larval stage increases the vector competence of Aedes aegypti females for Zika virus.

Aedes aegypti is the primary vector of Zika virus (ZIKV), a flavivirus which typically presents itself as febrile-like symptoms in humans but can also cause neurological and pregnancy complications. The transmission cycle of mosquito-borne arboviruses such as ZIKV requires that various key tissues in the female mosquito get productively infected with the virus before the mosquito can transmit the virus to another vertebrate host. Following ingestion of a viremic blood-meal from a vertebrate, ZIKV initially infects the midgut epithelium before exiting the midgut after blood-meal digestion to disseminate to secondary tissues including the salivary glands. Here we investigated whether smaller Ae. aegypti females resulting from food deprivation as larvae exhibited an altered vector competence for blood-meal acquired ZIKV relative to larger mosquitoes. Midguts from small 'Starve' and large 'Control' Ae. aegypti were dissected to visualize by transmission electron microscopy (TEM) the midgut basal lamina (BL) as physical evidence for the midgut escape barrier showing Starve mosquitoes with a significantly thinner midgut BL than Control mosquitoes at two timepoints. ZIKV replication was inhibited in Starve mosquitoes following intrathoracic injection of virus, however, Starve mosquitoes exhibited a significantly higher midgut escape and population dissemination rate at 9 days post-infection (dpi) via blood-meal, with more virus present in saliva and head tissue than Control by 10 dpi and 14 dpi, respectively. These results indicate that Ae. aegypti developing under stressful conditions potentially exhibit higher midgut infection and dissemination rates for ZIKV as adults, Thus, variation in food intake as larvae is potentially a source for variable vector competence levels of the emerged adults for the virus.

Ovitrap surveillance of dengue vector mosquitoes in Bandung City, West Java Province, Indonesia.

Larval surveillance is the central approach for monitoring dengue vector populations in Indonesia. However, traditional larval indices are ineffective for measuring mosquito population dynamics and predicting the dengue transmission risk. We conducted a 14-month ovitrap surveillance. Eggs and immature mosquitoes were collected on a weekly basis from an urban village of Bandung, namely Sekejati. Ovitrap-related indices, namely positive house index (PHI), ovitrap index (OI), and ovitrap density index (ODI), were generated and correlated with environmental variables, housing type (terraced or high-density housing), ovitrap placement location (indoor or outdoor; household or public place), and local dengue cases. Our results demonstrated that Aedes aegypti was significantly predominant compared with Aedes albopictus at each housing type and ovitrap placement location. Ovitrap placement locations and rainfall were the major factors contributing to variations in PHI, OI, and ODI, whereas the influences of housing type and temperature were subtle. Indoor site values were significantly positively correlated to outdoor sites' values for both OI and ODI. OI and ODI values from households were best predicted with those from public places at 1- and 0-week lags, respectively. Weekly rainfall values at 4- and 3-week lags were the best predictors of OI and ODI for households and public places, respectively. Monthly mean PHI, OI, and ODI were significantly associated with local dengue cases. In conclusion, ovitrap may be an effective tool for monitoring the population dynamics of Aedes mosquitoes, predicting dengue outbreaks, and serving as an early indicator to initiate environmental clean-up. Ovitrap surveillance is easy for surveyors if they are tasked with a certain number of ovitraps at a designated area, unlike the existing larval surveillance methodology, which entails identifying potential breeding sites largely at the surveyors' discretion. Ovitrap surveillance may reduce the influence of individual effort in larval surveillance that likely causes inconsistency in results.

Photodynamic control of Aedes aegypti larvae with environmentally-friendly tetra-platinated porphyrin.

This work evaluated the photosensitizing activity of isomeric tetra-cationic porphyrins with peripheral [Pt(bpy)Cl]+ to control the larval population of Aedes aegypti by photodynamic action. The photolarvicidal activity of the tetra-platinated porphyrins at meta and para position (3-PtTPyP and 4-PtTPyP) was evaluated under blue (450 nm), green (525 nm), and red (625 nm) light illumination at 55.0 J cm-2. The meta isomer presented an efficient photolarvicidal activity even at a low concentration (1.2 ppm) in the presence of light, while the para counterpart was inactive regardless of the concentration and illumination. The different responses were related to the improved optical features and higher water solubility of 3-PtTPyP compared to 4-PtTPyP. Additionally, the potential environmental toxicity of 3-PtTPyP was tested in a plant model (Allium cepa test), with no toxicity detected for all used concentrations (1.2 to 12 ppm). Hence, this work reveals that 3-PtTPyP has a great potential to be employed to photodynamically control the insect vector population in an environmentally safe way.