Identification of Three Novel Genes in Phenuiviridae Detected from Aedes Mosquitoes in Hokkaido, Japan.

Mosquitoes are important arthropod vectors of arboviruses. The family Phenuiviridae includes several medically important arboviruses, such as the Rift Valley fever phlebovirus and Toscana phlebovirus. Recent comprehensive genetic analyses have identified many novel mosquito-specific viruses that are phylogenetically related to Phenuiviridae. We collected mosquitoes from Hokkaido in northern Japan, and conducted reverse transcription polymerase chain reactions (RT-PCRs) targeting the RNA-dependent RNA polymerase (RdRp) gene of Phenuiviridae. A total of 285 pools, comprising 3,082 mosquitoes from 2 genera and 8 species, were collected. Partial RdRp sequences were detected in 97 pools, which allowed us to classify the viruses into 3 clusters provisionally designated as Etutanne virus (ETTV) 1, 2, and 3. The virus most closely related to ETTVs is Narangue virus (family Phenuiviridae, genus Mobuvirus), which was detected in Mansonia mosquitoes; the nucleotide and amino acid sequences of the Narangue virus are 58.4-66.2% and 64.7-86.7% similar, respectively, to those of ETTVs. PCR and RT-PCR using DNA and RNase digestion methods showed that the ETTVs are RNA viruses that do not form non-retroviral integrated RNA virus sequences in the mosquito genome.

Flower Mimics Roll Out Multicolored Carpets to Lure and Kill the House Fly.

Flowers and their spatial clustering are important parameters that mediate the foraging behavior and visitation rate of pollinating insects. Visual stimuli are crucial for triggering behavioral changes in the house fly, Musca domestica, which regularly visits plants for feeding and reproduction. The success of bait technology, which is the principal means of combatting flies, is adversely affected by reduced attractiveness and ineffective application techniques. Despite evidence that house flies have color vision capacity, respond to flowers, and exhibit color and pattern preference, the potential of artificial flowers as attractive factors has not been explored. The present study was performed to investigate whether artificial floral designs can lure and kill house flies. Starved wild house flies were presented with equal opportunities to acquire sugar meals, to which boric acid had been added as a toxin, from one flower arrangement (blue-dominated design, BDD; yellow-dominated design, YDD; or pink-dominated design, PDD), and a non-toxic white design (WDD). We also allowed house flies to forage within an enclosure containing two non-toxic floral designs (WDDs). The differences in mortality between the two environments with and without toxicant were examined. The survival rate of Musca domestica was extremely high when WDDs containing non-toxic sugar sources were the only feeding sites available. When given an option to forage in an environment containing a BDD and a WDD, house flies showed a high mortality rate (76%) compared to their counterparts maintained in the WDD environment (2%). When kept in an enclosure containing one YDD and a WDD, flies showed a mortality rate of 88%; however, no mortality occurred among flies confined to a compound with a WDD pair. When provided an even chance of foraging in an enclosure containing a mixed pair of floral arrangements (PDD and WDD) and another with two WDDs, flies showed a higher mortality rate (78%) in the first environment. However, the maximum survival rate (100%) was seen in the WDD environment. Exposure to YDD tended to result in a greater mortality rate than with the two other floral designs. Mortality gradually increased with time among flies exposed to tested artificial floral designs. The results presented here clearly indicated that artificial flower arrangements with a toxic sugar reward were strikingly attractive for house flies when their preferred color (white) was present. These observations offer novel possibilities for future development of flower mimic-based house fly control.

Development of genus-specific universal primers for the detection of flaviviruses.

BACKGROUND: Flaviviruses are representative arboviruses carried by arthropods and/or vertebrates; these viruses can pose a public health concern in many countries. By contrast, it is known that a novel virus group called insect-specific flaviviruses (ISFs) also infects arthropods, although no such virus has yet been isolated from vertebrates. The characteristics of ISFs, which affect replication of human-pathogenic flaviviruses within co-infected mosquito cells or mosquitoes without affecting the mosquitoes themselves, mean that we should pay attention to both ISFs and human-pathogenic flaviviruses, despite the fact that ISFs appear not to be directly hazardous to human health. To assess the risk of diseases caused by flaviviruses, and to better understand their ecology, it is necessary to know the extent to which flaviviruses are harbored by arthropods. METHODS: We developed a novel universal primer for use in a PCR-based system to detect a broad range of flaviviruses. We then evaluated its performance. The utility of the novel primer pair was evaluated in a PCR assay using artificially synthesized oligonucleotides derived from a template viral genome sequence. The utility of the primer pair was also examined by reverse transcription PCR (RT-PCR) using cDNA templates prepared from virus-infected cells or crude supernatants prepared from virus-containing mosquito homogenates. RESULTS: The novel primer pair amplified the flavivirus NS5 sequence (artificially synthesized) in all samples tested (six species of flavivirus that can cause infectious diseases in humans, and flaviviruses harbored by insects). In addition, the novel primer pair detected viral genomes in cDNA templates prepared from mosquito cells infected with live flavivirus under different infectious conditions. Finally, the viral genome was detected with high sensitivity in crude supernatants prepared from pooled mosquito homogenates. CONCLUSION: This PCR system based on a novel primer pair makes it possible to detect arthropod-borne flaviviruses worldwide (the primer pair even detected viruses belonging to different genetic subgroups). As such, an assay based on this primer pair may help to improve public health and safety, as well as increase our understanding of flavivirus ecology.

Zika Virus Potential Vectors among Aedes Mosquitoes from Hokkaido, Northern Japan: Implications for Potential Emergence of Zika Disease.

The Zika virus (ZIKV) is a rapidly expanding mosquito-borne virus that causes febrile illness in humans. Aedes aegypti and Ae. albopictus are the primary ZIKV vectors; however, the potential vector competence of other Aedes mosquitoes distributed in northern Japan (Palearctic ecozone) are not yet known. In this study, the susceptibility to Zika virus infection of three Aedes mosquitoes distributed in the main city of the northern Japan and their capacities as vectors for ZIKV were evaluated. Field-collected mosquitoes were fed ad libitum an infectious blood meal containing the ZIKV PRVABC59. The Zika virus was detected in the abdomen of Ae. galloisi and Ae. japonicus at 2-10 days post infection (PI), and from the thorax and head of Ae. galloisi at 10 days PI, resulting in 17.6% and 5.9% infection rates, respectively. The Zika virus was not detected from Ae. punctor at any time. Some northern Japanese Aedes could be suspected as vectors of ZIKV but the risk may be low when compared with major ZIKV vectors.

New Insights into the Biology of the Emerging Tembusu Virus.

Reported for the first time in 1955 in Malaysia, Tembusu virus (TMUV) remained, for a long time, in the shadow of flaviviruses with human health importance such as dengue virus or Japanese encephalitis virus. However, since 2010 and the first large epidemic in duck farms in China, the threat of its emergence on a large scale in Asia or even its spillover into the human population is becoming more and more significant. This review aims to report current knowledge on TMUV from viral particle organization to the development of specific vaccines and therapeutics, with a particular focus on host-virus interactions.

Relación entre el perfil antropométrico y aptitud física con el equilibrio postural dinámico en surfistas / Relationship between anthropometric profile and physical aptitude with dynamic postural balance in surfers

Introducción: El surf es una disciplina que ha aumentado considerablemente la cantidad de adeptos que practican estedeporte. Las investigaciones se han orientado en describir las principales variables asociadas al rendimiento, pero no lasrelaciones que pueden existir entre ellas. Objetivo: Establecer la relación del equilibrio postural dinámico con respecto al perfil antropométrico y aptitud física ensurfistas. Material y método: Esta investigación es un estudio de diseño observacional y transversal, de tipo descriptivo-correlacional. La muestra incluyo 30 surfistas (8 mujeres y 22 hombres). Las variables del perfil antropométrico estudiadas fueron: masacorporal, estatura bípeda, índice de masa corporal (IMC), suma de pliegues, composición corporal y somatotipo. Además, seevaluó la aptitud física mediante pruebas indirectas. Los datos del perfil antropométrico y aptitud física se correlacionaroncon los resultados de la prueba de equilibrio postural dinámico (Y balance test). Resultados: Para la dirección anterior del Y balance test, las variables que resultaron significativas fueron género, peso corporal,estatura bípeda, suma de pliegues, masa adiposa y altura del salto de Sargent (R2 = 0,55). La dirección posteromedial del Ybalance test arrojó un modelo significativo que indica que el género, suma de pliegues y masa adiposa son las variables quepredicen conjuntamente el equilibrio postural dinámico (R2 = 0,30). Para la dirección posterolateral el modelo señala que elIMC, suma de pliegues, masa adiposa, mesomorfismo, ectomorfismo y rendimiento en la prueba sit and reach son las variablesque influyen en rendimiento de la prueba Y balance test (R2 = 0,55). Conclusión: Se establecieron modelos predictivos para determinar variables antropométricas y de la aptitud física que seríandeterminantes para el desempeño del equilibrio postural de un surfista.(AU)

Introduction: Surfing is a discipline that has considerably increased the number of followers who practice this sport. Re-search has focused on describing the main variables associated with performance, but not the relationships that may existbetween them. Objective: To establish the relationship of dynamic postural balance with respect to the anthropometric profile and physicalaptitude in surfers. Material and method: This research is an observational and cross-sectional study of a descriptive-correlational type. Thesample included 30 surfers (8 women and 22 men). The variables of the anthropometric profile studied were body mass,bipedal height, body mass index (BMI), sum of folds, body composition and somatotype. Furthermore, physical aptitude wasassessed by indirect tests. The data of the anthropometric profile and physical aptitude were correlated with the results ofthe dynamic postural balance test (Y balance test). Results: For the anterior direction of the Y balance test, the variables that were significant were gender, body weight, bipedalheight, sum of folds, adipose mass and Sargent’s jump height (R2 = 0.55). The posteromedial direction of the Y balance testyielded a significant model that indicates that gender, sum of folds and adipose mass are the variables that jointly predictdynamic postural balance (R2 = 0.30). For the posterolateral direction, the model indicates that the BMI, sum of folds, adiposemass, mesomorphism, ectomorphism and performance in the sit and reach test are the variables that influence the perfor-mance of the Y balance test (R2 = 0.55). Conclusion: Predictive models were established to determine anthropometric and physical condition variables that wouldbe decisive for the performance of a surfer’s postural balance.(AU)

Vector Competence for Dengue-2 Viruses Isolated from Patients with Different Disease Severity.

Dynamics of dengue serotype 2 virus isolated from patients with different disease severity, namely flu-like classic dengue fever (DF) and dengue shock syndrome (DSS) were studied in its mosquito vector Aedes aegypti. We compared isolate infectivity and vector competence (VC) among thirty two A. aegypti-viral isolate pairs. Mosquito populations from high dengue incidence area exhibited overall greater VC than those from low dengue incidence area at 58.1% and 52.5%, respectively. On the other hand, the overall infection rates for the isolates ThNR2/772 (DF, 62.3%) and ThNR2/391 (DSS, 60.9%), were significantly higher than those for isolates ThNR2/406 (DF, 55.2%) and ThNR2/479 (DSS, 54.8%). These results suggest that the efficacy of dengue virus circulation was likely to vary according to the combination between the virus strains and origin of the mosquito strains, and this may have epidemiologic implications toward the incidence of flu-like classic dengue fever (DF) and dengue shock syndrome (DSS).

Differential Susceptibility and Innate Immune Response of Aedes aegypti and Aedes albopictus to the Haitian Strain of the Mayaro Virus.

Mayaro (MAYV) is an emerging arthropod-borne virus belonging to the Alphavirus genus of the Togaviridae family. Although forest-dwelling Haemagogus mosquitoes have been considered as its main vector, the virus has also been detected in circulating Aedes ssp mosquitoes. Here we assess the susceptibility of Aedes aegypti and Aedes albopictus to infection with MAYV and their innate immune response at an early stage of infection. Aedes albopictus was more susceptible to infection with MAYV than Ae. aegypti. Analysis of transcript levels of twenty immunity-related genes by real-time PCR in the midgut of both mosquitoes infected with MAYV revealed increased expression of several immune genes, including CLIP-domain serine proteases, the anti-microbial peptides defensin A, E, cecropin E, and the virus inducible gene. The regulation of certain genes appeared to be Aedes species-dependent. Infection of Ae. aegypti with MAYV resulted in increased levels of myeloid differentiation2-related lipid recognition protein (ML26A) transcripts, as compared to Ae. albopictus. Increased expression levels of thio-ester-containing protein 22 (TEP22) and Niemann-Pick type C1 (NPC1) gene transcripts were observed in infected Ae. albopictus, but not Ae. aegypti. The differences in these gene expression levels during MAYV infection could explain the variation in susceptibility observed in both mosquito species.

Interferon-inducible protein (IFI) 16 regulates Chikungunya and Zika virus infection in human skin fibroblasts.

Chikungunya virus (CHIKV), a re-emerging infectious arbovirus, causes Chikungunya fever that is characterized by fever, skin rash, joint pain, arthralgia and occasionally death. Despite it has been described for 66 years already, neither potential vaccine nor a specific drug is available yet. During CHIKV infection, interferon type I signaling pathway is stimulated and releases hundreds of interferon stimulated genes (ISGs). Our previous study reported that IFI16, a member of ISGs, is up-regulated during CHIKV virus infection and the suppression of the gene resulted in increased virus replication. Furthermore, our group also found that inflammasome activation can inhibit CHIKV infection in human foreskin cells (HFF1). Concomitantly, it has been reported that IFI16 activates the inflammasome to suppress virus infection. Therefore, we have hypothesized that IFI16 could be involved in CHIKV infection. In this study, we confirmed the expression level of IFI16 by Western blotting analysis and found that IFI16 was up-regulated following CHIKV infection in both HFF1 and human embryonic kidney cells. We next investigated its antiviral activity and found that forced expression of IFI16 completely restricted CHIKV infection while endogenous silencing of the gene markedly increased virus replication. Furthermore, we have discovered that IFI16 inhibited CHIKV replication, at least, in cell-to-cell transmission as well as the diffusion step. Interestingly, IFI16 also exerted its antiviral activity against Zika virus (ZIKV) infection, the global threat re-emerging virus can cause microcephaly in humans. Taken together, this study provides the first evidence of an antivirus activity of IFI16 during in vitro arbovirus infection, thus expanding its antiviral spectrum that paves the way to further development of antiviral drugs and vaccines.

The electronic song "Scary Monsters and Nice Sprites" reduces host attack and mating success in the dengue vector Aedes aegypti.

Sound and its reception are crucial for reproduction, survival, and population maintenance of many animals. In insects, low-frequency vibrations facilitate sexual interactions, whereas noise disrupts the perception of signals from conspecifics and hosts. Despite evidence that mosquitoes respond to sound frequencies beyond fundamental ranges, including songs, and that males and females need to struggle to harmonize their flight tones, the behavioral impacts of music as control targets remain unexplored. In this study, we examined the effects of electronic music (Scary Monsters and Nice Sprites by Skrillex) on foraging, host attack, and sexual activities of the dengue vector Aedes aegypti. Adults were presented with two sound environments (music-off or music-on). Discrepancies in visitation, blood feeding, and copulation patterns were compared between environments with and without music. Ae. aegypti females maintained in the music-off environment initiated host visits earlier than those in the music-on environment. They visited the host significantly less often in the music-on than the music-off condition. Females exposed to music attacked hosts much later than their non-exposed peers. The occurrence of blood feeding activity was lower when music was being played. Adults exposed to music copulated far less often than their counterparts kept in an environment where there was no music. In addition to providing insight into the auditory sensitivity of Ae. aegypti to sound, our results indicated the vulnerability of its key vectorial capacity traits to electronic music. The observation that such music can delay host attack, reduce blood feeding, and disrupt mating provides new avenues for the development of music-based personal protective and control measures against Aedes-borne diseases.

Geometric morphometrics approach towards discrimination of three member species of Maculatus group in Thailand.

Members of the Maculatus group are important malaria vectors in the border regions of Thailand. However, the role of each species in malaria transmission remains unclear because of their highly similar morphologies, making them difficult to be differentiated. Whereas An. pseudowillmori may be identified by the color pattern of some scales on abdomen and wings, the distinction between An. maculatus and An. sawadwongporni relies on the wings only. Scales are labile structures, as they may be accidentally removed during capture and transportation to the laboratory. To discriminate among the species of this group, we tested the suitability of geometric techniques. Shape variables were used as input for discriminant analyses and validated reclassification. Both landmark- and outline-based geometric techniques disclosed significant differences between the three species. For the delicate An. maculatus - An. sawadwongporni distinction, the outline-based approach appeared as the most promising, with validated reclassification scores reaching 93%, as compared to 77% obtained by landmark data. For An. pseudowillmori, reclassification scores were 100% and 94%, respectively. Geometric morphometrics may provide an alternative and useful complement for discriminating members of the Maculatus group.

Courtship activity, copulation & insemination success in a mosquito vector fed a herbal aphrodisiac: Implications for sterile insect technology.

BACKGROUND & OBJECTIVES: In sterile insect technology (SIT), mating competitiveness is a pre-condition for the reduction of target pest populations and a crucial parameter for judging efficacy. Still, current SIT trials are being hindered by decreased effectiveness due to reduced sexual performance of released males. Here, we explored the possible role of a herbal aphrodisiac in boosting the mating activity of Aedes aegypti. METHODS: Males were fed one of two diets in this study: experimental extract of Eurycoma longifolia (MSAs) and sugar only (MSOs). Differences in life span, courtship latency, copulation activity and mating success were examined between the two groups. RESULTS: No deaths occurred among MSA and MSO males. Life span of MSOs was similar to that of MSAs. The courtship latency of MSAs was shorter than that of MSOs (P<0.01). MSAs had greater copulation success than MSOs (P<0.001). In all female treatments, MSAs mated more than MSOs, but the differences in rate were significant only in the highest female density (P<0.05). In MSAs, mating success varied significantly with female density (P<0.01), with the 20-female group (P<0.01) having the lowest rate. Single MSA had better mating success at the two lowest female densities. In MSOs, there were no significant differences in mating success rate between the different female densities. INTERPRETATION & CONCLUSIONS: Our results suggested that the herbal aphrodisiac, E. longifolia, stimulated the sexual activity of Ae. aegypti and may be useful for improving the mating competitiveness of sterile males, thus improving SIT programmes.

Exposure of a diurnal mosquito vector to floral mimics: Foraging responses, feeding patterns, and significance for sugar bait technology.

Food location by mosquitoes is mediated by resource-derived olfactory and visual signals. Smell sensation is intermittent and dependent on the environment, whereas visual signals are continual and precede olfactory cues. Success of mosquito bait technology, where olfactory cues are used for attraction, is being impeded by reduced attractiveness. Despite proof that mosquitoes respond to colored objects, including those mimicking floral shape, and that they can discriminate among flowers, the impacts of artificial flowers on foraging remain unexplored. Using artificial flowers with sugar rewards, we examined the foraging responses of Aedes aegypti to various colors in equal choice bioassays. Starved adults were exposed to single flowers with petals of a given color (Single Blue Flowers [SBFs]; Single Red Flowers [SRFs]; Single Yellow Flowers [SYFs]; Single Pink Flowers [SPIFs]; and Single Purple Flowers [SPFs]) and two others with white petals (SWFs). Discrepancies in response time, visitation, feeding, and resting of both sexes were compared between colored flowers and SWFs. Ae. aegypti exhibited shorter response times to colored flowers compared to SWFs, but this behavior was mostly seen for SBFs or SYFs in females, and SRFs, SYFs, SPIFs, or SPFs in males. When provided an option to land on colored flowers and SWFs, female visitation occurred at high rates on SBFs, SRFs, SYFs, SPIFs, and SPFs; for males, this preference for colored flowers was seen to a lesser degree on SBF and SPIFs. Both sexes exhibited preference for colored flowers as sugar sources, but with different patterns: SPIFs, SRFs, SYFs, and SPFs for females; SYFs, SPFs, SPIFs and SRFs for males. Females preferentially rested on colored flowers when in competition with SWFs, but this preference was more pronounced for SPFs, SRFs, and SBFs. Males exhibited an increased preference for SRFs, SPFs, and SYFs as resting sites. Our results indicated the attraction of Ae. aegypti to rewarding artificial flowers, in some cases in ways similar to live flowering plants. The discovery that both male and female Ae. aegypti can feed on nectar mimics held by artificial flowers opens new avenues for improving sugar bait technology and for developing new attract-and-kill devices.

Occurrence of sweet refuse at disposal sites: rainwater retention capacity and potential breeding opportunities for Aedes aegypti.

Nectar is the staple diet of adult mosquitoes in the wild, but its availability is inconsistent and can be affected by rainfall. In urban centers, Aedes vectors commonly use man-made containers as their major habitat; however, they can colonize any items replenished by rainfall. Garbage output has increased significantly in recent years, at a time when collection frequency is reducing. Such garbage usually includes organic components, some of which are sweet and can be fed upon by other animals or become can containers for rainwater. Despite evidence that Aedes larvae can thrive in containers comprised of organic waste material, which can be produced by rodents gnawing on fruits or vegetables, and that adults can survive on sweet waste fluids, the capacity of organic waste materials to accumulate rainwater and act as egg deposition sites has not been examined. It is also unknown for how long sweet extracts can sustain the life of adult vectors. Here, we investigated the abundance of sweet leftovers at garbage sites and the rainwater retention capacity of some organic materials through a field survey and laboratory bioassays. We also examined whether sweet waste fluids impact egg hatching success and longevity of Aedes aegypti. The results of this study indicated that sweet products with leftovers are highly prevalent in garbage. When exposed to rain, food items (BAFrc, banana fruit resembling container; and BSPrc, boiled sweet potato resembling container) and the packaging of sweet foods (SMIc, sweetened condensed milk can) retained water. When provided an opportunity to oviposit in cups containing BAF extract (BAFex), BSP extract (BSPex), and SMI extract (SMIex), eggs were deposited in all media. Egg maturation in the BAFex environment resulted in similar larval eclosion success to that resulting from embryo development in a water milieu. Adults maintained on sweet waste extracts had long lifespans, although shorter than that of their sugar solution (SUS)-fed counterparts. Taken together, these results indicated that sweet waste materials are useful to dengue mosquitoes, acting both as oviposition sites and energy sources.

Co-Infection of Mosquitoes with Chikungunya and Dengue Viruses Reveals Modulation of the Replication of Both Viruses in Midguts and Salivary Glands of Aedes aegypti Mosquitoes.

Arthropod-borne virus (arbovirus) infections cause several emerging and resurgent infectious diseases in humans and animals. Chikungunya-affected areas often overlap with dengue-endemic areas. Concurrent dengue virus (DENV) and chikungunya virus (CHIKV) infections have been detected in travelers returning from regions of endemicity. CHIKV and DENV co-infected Aedes albopictus have also been collected in the vicinity of co-infected human cases, emphasizing the need to study co-infections in mosquitoes. We thus aimed to study the pathogen-pathogen interaction involved in these co-infections in DENV/CHIKV co-infected Aedes aegypti mosquitoes. In mono-infections, we detected CHIKV antigens as early as 4 days post-virus exposure in both the midgut (MG) and salivary gland (SG), whereas we detected DENV serotype 2 (DENV-2) antigens from day 5 post-virus exposure in MG and day 10 post-virus exposure in SG. Identical infection rates were observed for singly and co-infected mosquitoes, and facilitation of the replication of both viruses at various times post-viral exposure. We observed a higher replication for DENV-2 in SG of co-infected mosquitoes. We showed that mixed CHIKV and DENV infection facilitated viral replication in Ae. aegypti. The outcome of these mixed infections must be further studied to increase our understanding of pathogen-pathogen interactions in host cells.

Anti-oviposition activities of used sock media against a dengue vector: prospects of eco-friendly control and solutions to pollution.

Yearly, huge amounts of sock refuse are discarded into the environment. Socks contain many molecules, and worn ones, which are rich in smell-causing bacteria, have a strong influence on animals' behaviors. But the impacts of sock odor on the oviposition behavior of dengue vectors are unknown. We assessed whether Aedes albopictus changes its oviposition activity in response to the presence of used socks extract (USEx) in potential breeding grounds, using choice and no-choice bioassays (NCB). When furnished even chances to oviposit in two sites holding USEx and two others containing water (control), Ae. albopictus deposited significantly less eggs in USEx than in water sites. A similar pattern of oviposition preference was also observed when there were more oviposition options in water. When there were greater oviposition opportunities in USEx sites, Ae. albopictus oviposited preferentially in water. Females laid significantly more eggs during the NCB involving water than USEx. Also, significantly more mature eggs were retained by females in the NCB with USEx than in that with water. These observations strongly suggest the presence of molecules with either repellent or deterrent activities against Ae. albopictus females and provide an impetus to advocate the integration of used socks in dengue control programs. Such applications could be a realistic end-of-life recourse to reroute this waste from landfills.

Coffee, its roasted form, and their residues cause birth failure and shorten lifespan in dengue vectors.

In dengue mosquitoes, successful embryonic development and long lifespan are key determinants for the persistence of both virus and vector. Therefore, targeting the egg stage and vector lifespan would be expected to have greater impacts than larvicides or adulticides, both strategies that have lost effectiveness due to the development of resistance. Therefore, there is now a pressing need to find novel chemical means of vector control. Coffee contains many chemicals, and its waste, which has become a growing environmental concern, is as rich in toxicants as the green coffee beans; these chemicals do not have a history of resistance in insects, but some are lost in the roasting process. We examined whether exposure to coffee during embryonic development could alter larval eclosion and lifespan of dengue vectors. A series of bioassays with different coffee forms and their residues indicated that larval eclosion responses of Aedes albopictus and Ae. aegypti were appreciably lower when embryonic maturation occurred in environments containing coffee, especially roasted coffee crude extract (RCC). In addition, the lifespan of adults derived from eggs that hatched successfully in a coffee milieu was reduced, but this effect was less pronounced with roasted and green coffee extracts (RCU and GCU, respectively). Taken together, these findings suggested that coffee and its residues have embryocidal activities with impacts that are carried over onto the adult lifespan of dengue vectors. These effects may significantly reduce the vectorial capacity of these insects. Reutilizing coffee waste in vector control may also represent a realistic solution to the issues associated with its pollution.

A Zika virus from America is more efficiently transmitted than an Asian virus by Aedes aegypti mosquitoes from Asia.

Zika is a mosquito-borne disease associated with neurological disorders that causes an on-going pandemic. The first outbreak was recorded in Micronesia in 2007, then in French Polynesia in 2014 from which it spread to South America in 2015 and ignited a widespread epidemic. Interestingly, Zika outbreaks in Asia remained of moderate intensity although the virus is circulating. To understand these epidemiological variations, we investigated the entomological determinants of ZIKV transmission in Asia. We used oral infection of mosquitoes collected in Singapore to identify the vector species, to quantify the blood infection threshold and to compare transmissibility between an Asian ZIKV strain (H/PF13) and an American strain collected in Brazil (BE H 815744). We have confirmed the vector status of Aedes aegypti and determined that 103 pfu/ml of blood is sufficient to infect mosquitoes. We showed that only the American strain was present in the saliva 3 days post-infection, and that this strain had a 30-40% higher rate of saliva infection in Ae. aegypti from 3 to 14 days post-infection than the Asian strain. Our data suggests that American strains are more efficiently transmitted than Asian strains, which raises concerns about the introduction of American strains in Asia.

Sweet waste extract uptake by a mosquito vector: Survival, biting, fecundity responses, and potential epidemiological significance.

In nature, adult mosquitoes typically utilize nectar as their main energy source, but they can switch to other as yet unidentified sugary fluids. Contemporary lifestyles, with their associated unwillingness to consume leftovers and improper disposal of waste, have resulted in the disposal of huge amounts of waste into the environment. Such refuse often contains unfinished food items, many of which contain sugar and some of which can collect water from rain and generate juices. Despite evidence that mosquitoes can feed on sugar-rich suspensions, semi-liquids, and decaying fruits, which can be abundant in garbage sites, the impacts of sweet waste fluids on dengue vectors are unknown. Here, we investigated the effects of extracts from some familiar sweet home waste items on key components of vectorial capacity of Aedes aegypti. Adult mosquitoes were fed one of five diets in this study: water (WAT); sucrose (SUG); bakery product (remnant of chocolate cake, BAK); dairy product (yogurt, YOG); and fruit (banana (BAN). Differences in survival, response time to host, and egg production were examined between groups. For both males and females, maintenance on BAK extract resulted in marked survival levels that were similar to those seen with SUG. Sweet waste extracts provided better substrates for survival compared to water, but this superiority was mostly seen with BAK. Females maintained on BAK, YOG, and BAN exhibited shorter response times to a host compared to their counterparts maintained on SUG. The levels of egg production were equivalent in waste extract- and SUG-fed females. The findings presented here illustrate the potential of sweet waste-derived fluids to contribute to the vectorial capacity of dengue vectors and suggest the necessity of readdressing the issue of waste disposal, especially that of unfinished sweet foods. Such approaches can be particularly relevant in dengue endemic areas where rainfall is frequent and waste collection infrequent.