bims-mosdis Biomed News
on Mosquito distribution and disease
Issue of 2020‒05‒24
thirty papers selected by
Richard Halfpenny
Staffordshire University


  1. Insects. 2020 May 14. pii: E307. [Epub ahead of print]11(5):
    Tuno N, Phong TV, Takagi M.
      (1) Dengue is the most spread mosquito-borne viral disease in the world, and vector control is the only available means to suppress its prevalence, since no effective treatment or vaccine has been developed. A biological control program using copepods that feed on mosquito larvae has been practiced in Vietnam and some other countries, but the application of copepods was not always successful. (2) To understand why the utility of copepods varies, we evaluated the predation efficiency of a copepod species (Mesocyclops aspericornis) on a vector species (Aedes aegypti) by laboratory experiments under different temperatures, nutrition and prey-density conditions. (3) We found that copepod predation reduced intraspecific competition among Aedes larvae and then shortened the survivor's aquatic life and increased their pupal weight. In addition, the predatory efficiency of copepods was reduced at high temperatures. Furthermore, performance of copepod offspring fell when the density of mosquito larvae was high, probably because mosquito larvae had adverse effects on copepod growth through competition for food resources. (4) These results suggest that the increase in mosquitoes will not be suppressed solely by the application of copepods if the density of mosquito larvae is high or ambient temperature is high. We need to consider additional control methods in order to maintain the efficiency of copepods to suppress mosquito increase.
    Keywords:  Aedes aegypti; biological control; copepod; dengue fever
    DOI:  https://doi.org/10.3390/insects11050307
  2. Parasit Vectors. 2020 May 20. 13(1): 264
    Hayd RLN, Carrara L, de Melo Lima J, de Almeida NCV, Lima JBP, Martins AJ.
      BACKGROUND: Roraima, the northernmost State in Brazil, borders Venezuela and Guyana. Although mostly covered by the tropical forests, the urban centers of this state are highly infested with Ae. aegypti and are endemic for dengue, Zika and chikungunya. We accessed the insecticide resistance status of Ae. aegypti populations from the capital Boa Vista, two cities on international borders (Pacaraima and Bonfim) and Rorainópolis bordering Amazonas State, in order to evaluate the chemical control efficacy in these localities.METHODS: Tests with World Health Organization (WHO)-like tubes impregnated with the pyrethroid deltamethrin (0.05% and 0.12%) and the organophosphate malathion (0.7%) were conducted with Ae. aegypti from Boa Vista, Pacaraima, Bonfim and Rorainópolis, collected in 2016 and 2018. Genotyping of kdr mutations, related to resistance to pyrethroids, was performed for the SNP variations at sites 1016 and 1534 of the voltage gated sodium channel gene (NaV) with a TaqMan qPCR approach.
    RESULTS: Aedes albopictus was absent in our collections, and therefore only Ae. aegypti was tested. All Ae. aegypti populations were susceptible to 0.7% malathion in 2016; however, mortality dropped to under 90% in Bonfim and Pacaraima populations in 2018. All populations were resistant to 0.05% deltamethrin in both years. The time that 50% of females suffered knockdown (KdT50) under exposure to 0.05% deltamethrin was 3.3-5.9-fold longer in mosquitoes from the natural populations compared to the susceptible Rockefeller strain. Only the Pacaraima population (2018) remained resistant to 0.12% deltamethrin. Kdr genotyping revealed the absence of the wild-type NaVS haplotype (1016Val + 1534Phe) in the populations from Roraima, indicating that all tested insects had a genetic background for pyrethroid resistance. The double kdr NaVR2 haplotype (1016Ile + 15434Cys) was present in higher frequencies in all populations except for Rorainópolis, where this haplotype seems to have arrived recently.
    CONCLUSIONS: These results are important for the knowledge about insecticide resistance status of Ae. aegypti populations from Roraima and will help improve vector control strategies that may be applied to diverse localities under similar geographical and urban conditions.
    Keywords:  Boa Vista; Bonfim; Deltamethrin; Insecticide resistance monitoring; Malathion; Pacaraima; Rorainópolis; Vector control
    DOI:  https://doi.org/10.1186/s13071-020-04127-w
  3. Insects. 2020 May 15. pii: E312. [Epub ahead of print]11(5):
    Mayi MPA, Bamou R, Djiappi-Tchamen B, Fontaine A, Jeffries CL, Walker T, Antonio-Nkondjio C, Cornel AJ, Tchuinkam T.
      To identify potential sylvatic, urban and bridge-vectors that can be involved in current or future virus spillover from wild to more urbanised areas, entomological field surveys were conducted in rural, peri-urban and urban areas spanning the rainy and dry seasons in western Cameroon. A total of 2650 mosquitoes belonging to 37 species and eight genera were collected. Mosquito species richness was significantly influenced by the specific combination of the habitat type and the season. The highest species richness was found in the peri-urban area (S = 30, Chao1 = 121 ± 50.63, ACE = 51.97 ± 3.88) during the dry season (S = 28, Chao1 = 64 ± 25.7, ACE = 38.33 ± 3.1). Aedes (Ae.) africanus and Culex (Cx.) moucheti were only found in the rural and peri-urban areas, while Cx. pipiens s.l. and Ae. aegypti were only found in the urban area. Cx. (Culiciomyia) spp., Cx. duttoni and Ae. albopictus were caught in the three habitat types. Importantly, approximately 52% of the mosquito species collected in this study have been implicated in the transmission of diverse arboviruses. This entomological survey provides a catalogue of the different mosquito species that may be involved in the transmission of arboviruses. Further investigations are needed to study the vectorial capacity of each mosquito species in arbovirus transmission.
    Keywords:  Cameroon; Dschang; arboviruses; emerging vector-borne diseases; mosquito-vectors; urbanisation
    DOI:  https://doi.org/10.3390/insects11050312
  4. Pathogens. 2020 May 15. pii: E380. [Epub ahead of print]9(5):
    Mancini MV, Damiani C, Short SM, Cappelli A, Ulissi U, Capone A, Serrao A, Rossi P, Amici A, Kalogris C, Dimopoulos G, Ricci I, Favia G.
      Mosquitoes can transmit many infectious diseases, such as malaria, dengue, Zika, yellow fever, and lymphatic filariasis. Current mosquito control strategies are failing to reduce the severity of outbreaks that still cause high human morbidity and mortality worldwide. Great expectations have been placed on genetic control methods. Among other methods, genetic modification of the bacteria colonizing different mosquito species and expressing anti-pathogen molecules may represent an innovative tool to combat mosquito-borne diseases. Nevertheless, this emerging approach, known as paratransgenesis, requires a detailed understanding of the mosquito microbiota and an accurate characterization of selected bacteria candidates. The acetic acid bacteria Asaia is a promising candidate for paratransgenic approaches. We have previously reported that Asaia symbionts play a beneficial role in the normal development of Anopheles mosquito larvae, but no study has yet investigated the role(s) of Asaia in adult mosquito biology. Here we report evidence on how treatment with a highly specific anti-Asaia monoclonal antibody impacts the survival and physiology of adult Anopheles stephensi mosquitoes. Our findings offer useful insight on the role of Asaia in several physiological systems of adult mosquitoes, where the influence differs between males and females.
    Keywords:  Anopheles; Asaia; symbiont
    DOI:  https://doi.org/10.3390/pathogens9050380
  5. Arch Virol. 2020 May 21.
    Martin E, Tang W, Briggs C, Hopson H, Juarez JG, Garcia-Luna SM, de Valdez MW, Badillo-Vargas IE, Borucki MK, Frank M, Hamer GL.
      South Texas has experienced local transmission of Zika virus and of other mosquito-borne viruses such as chikungunya virus and dengue virus in the last decades. Using a mosquito surveillance program in the Lower Rio Grande Valley (LRGV) and San Antonio, TX, from 2016 to 2018, we detected the presence of an insect-specific virus, cell fusing agent virus (CFAV), in the Aedes aegypti mosquito population. We tested 6,326 females and 1,249 males from the LRGV and 659 females from San Antonio for CFAV by RT-PCR using specific primers. Infection rates varied from 0 to 261 per 1,000 mosquitoes in the LRGV and 115 to 208 per 1,000 in San Antonio depending on the month of collection. Infection rates per 1,000 individuals appeared higher in females collected from BG Sentinel 2 traps compared to Autocidal Gravid Ovitraps, but the ratio of the percentage of infected pools did not differ by trap type. The natural viral load in individual males ranged from 1.25 x 102 to 5.50 x 106 RNA copies and in unfed females from 5.42 x 103 to 8.70 x 106 RNA copies. Gravid females were found to harbor fewer viral particles than males and unfed females.
    DOI:  https://doi.org/10.1007/s00705-020-04652-0
  6. Parasit Vectors. 2020 May 16. 13(1): 259
    Gowelo S, Chirombo J, Spitzen J, Koenraadt CJM, Mzilahowa T, van den Berg H, Takken W, McCann R.
      BACKGROUND: Application of the larvicide Bacillus thuringiensis var. israelensis (Bti) is a viable complementary strategy for malaria control. Efficacy of Bti is dose-dependent. There is a knowledge gap on the effects of larval exposure to sublethal Bti doses on emerging adult mosquitoes. The present study examined the effect of larval exposure to sublethal doses of Bti on the survival, body size and oviposition rate in adult Anopheles coluzzii.METHODS: Third-instar An. coluzzii larvae were exposed to control and sublethal Bti concentrations at LC20, LC50 and LC70 for 48 h. Surviving larvae were reared to adults under standard colony conditions. Thirty randomly selected females from each treatment were placed in separate cages and allowed to blood feed. Twenty-five gravid females from the blood-feeding cages were randomly selected and transferred into new cages where they were provided with oviposition cups. Numbers of eggs laid in each cage and mortality of all adult mosquitoes were recorded daily. Wing lengths were measured of 570 mosquitoes as a proxy for body size.
    RESULTS: Exposure to LC70Bti doses for 48 h as third-instar larvae reduced longevity of adult An. coluzzii mosquitoes. Time to death was 2.58 times shorter in females exposed to LC70Bti when compared to the control females. Estimated mortality hazard rates were also higher in females exposed to the LC50 and LC20 treatments, but these differences were not statistically significant. The females exposed to LC70 concentrations had 12% longer wings than the control group (P < 0.01). No differences in oviposition rate of the gravid females were observed between the treatments.
    CONCLUSIONS: Exposure of An. coluzzii larvae to sublethal Bti doses reduces longevity of resultant adults and is associated with larger adult size and unclear effect on oviposition. These findings suggest that anopheline larval exposure to sublethal Bti doses, though not recommended, could reduce vectorial capacity for malaria vector populations by increasing mortality of resultant adults.
    Keywords:  Bacillus thuringiensis var. israelensis; Larval source management; Mosquito; Sublethal dose; Vector control
    DOI:  https://doi.org/10.1186/s13071-020-04132-z
  7. Vet Parasitol. 2020 May 01. pii: S0304-4017(20)30085-6. [Epub ahead of print]282 109105
    Todorovic S, McKay T.
      The nematode Dirofilaria immitis causes serious disease of canines in the United States. Transmitted by a variety of mosquito species, several studies have examined the prevalence of D. immitis in mosquitoes. However, no prevalence studies have focused on diurnal mosquitoes that could potentially enter homes. Mosquitoes were collected under doorway awnings at four residences to identify the diurnal blood-feeding mosquito species and prevalence of potential vectors harboring D. immitis in Jonesboro, Arkansas. Mosquitoes were collected from May to December 2016 using pheromone traps and aerial nets. Each mosquito was individually dissected and observed for larvae in the head, thorax, and abdomen to identify infective and non-infective larvae. Ten mosquito species from five genera were identified. Larvae, presumed to be D. immitis, were observed in five mosquito species, including Aedes albopictus, Anopheles quadrimaculatus, Culex pipiens quinquefasciatus, Culiseta inornata, and Psorophora columbiae and four genera, including Anopheles spp., Culex spp., Culiseta spp., and Psorophora spp. The diurnal mosquito, Ae. albopictus composed 3.4% of the total mosquitoes collected. An. quadrimaculatus and Anopheles spp. were the most abundant mosquitoes, composing 59.7 and 19.6% of the total mosquitoes collected, respectively. Of the 536 mosquitoes dissected, 60 (11.2%) were observed with D. immitis, and 33 (55.0%) of the heartworm-positive mosquitoes harbored infective (L3) larvae in the head or proboscis upon dissection. The high proportion of infective D. immitis larvae found in the mosquitoes aggregating under doorway awnings may increase the exposure of indoor dogs and cats to heartworm disease in northeast Arkansas.
    Keywords:  Heartworm; Mosquitoes
    DOI:  https://doi.org/10.1016/j.vetpar.2020.109105
  8. One Health. 2020 Dec;10 100138
    Ahmed MAI, Vogel CFA.
      Synergistic effects of octopamine receptor agonists (OR agonists) have attracted many scientists based on their potent effects on mosquitoes. Herein, we determined the toxicity of selected insect growth regulators (IGRs) on fourth instar larvae of Culex quinquefasciatus. We evaluated the synergistic action of OR agonists on the toxicity of IGR insecticides to achieve a better understanding of their mode of action. As a result, pyriproxyfen was the most potent IGR insecticide (EC50 = 0.049 ng/ml) followed by lufenuron, novaluron, and diflubenzuron according to the IGR bioassay. Further, based on the acute bioassay, lufenuron was the most toxic IGR insecticide (LC50 = 44 ng/ml) after 24-h post treatment followed by pyriproxyfen, novaluron, and diflubenzuron (LC50 = 137, 263, and 1127 ng/ml, respectively). Similar tendency was observed after 48 and 72-h post treatment. Furthermore, OR agonists that combined with pyriproxyfen was the most significant effects after 48 and 72-h of exposure. The synergism with amitraz (AMZ) was more significant when co-treated with IGR insecticides compared to chlordimeform (CDM). These findings suggest that OR agonists are promising tools and are important alternative strategies as synergistic compounds in preventing and controlling Culex quinquefasciatus mosquitoes.
    Keywords:  Culex quinquefasciatus; IGRs; Insecticide resistance; Mosquito control; Octopamine receptor agonists (OR agonists)
    DOI:  https://doi.org/10.1016/j.onehlt.2020.100138
  9. Environ Health Perspect. 2020 May;128(5): 57007
    Khan SU, Ogden NH, Fazil AA, Gachon PH, Dueymes GU, Greer AL, Ng V.
      BACKGROUND: Aedes aegypti and Ae. albopictus are mosquito vectors of more than 22 arboviruses that infect humans.OBJECTIVES: Our objective was to develop regional ecological niche models for Ae. aegypti and Ae. albopictus in the conterminous United States and Canada with current observed and simulated climate and land-use data using boosted regression trees (BRTs).
    METHODS: We used BRTs to assess climatic suitability for Ae. albopictus and Ae. aegypti mosquitoes in Canada and the United States under current and future projected climates.
    RESULTS: Models for both species were mostly influenced by minimum daily temperature and demonstrated high accuracy for predicting their geographic ranges under the current climate. The northward range expansion of suitable niches for both species was projected under future climate models. Much of the United States and parts of southern Canada are projected to be suitable for both species by 2100, with Ae. albopictus projected to expand its range north earlier this century and further north than Ae. aegypti.
    DISCUSSION: Our projections suggest that the suitable ecological niche for Aedes will expand with climate change in Canada and the United States, thus increasing the risk of Aedes-transmitted arboviruses. Increased surveillance for these vectors and the pathogens they carry would be prudent. https://doi.org/10.1289/EHP5899.
    DOI:  https://doi.org/10.1289/EHP5899
  10. Int J Environ Res Public Health. 2020 May 15. pii: E3450. [Epub ahead of print]17(10):
    Ibáñez-Justicia A, Smitz N, den Hartog W, van de Vossenberg B, De Wolf K, Deblauwe I, Van Bortel W, Jacobs F, Vaux AGC, Medlock JM, Stroo A.
      In Europe, the air-borne accidental introduction of exotic mosquito species (EMS) has been demonstrated using mosquito surveillance schemes at Schiphol International Airport (Amsterdam, The Netherlands). Based upon these findings and given the increasing volume of air transport movements per year, the establishment of EMS after introduction via aircraft is being considered a potential risk. Here we present the airport surveillance results performed by the Centre for Monitoring of Vectors of the Netherlands, by the Monitoring of Exotic Mosquitoes (MEMO) project in Belgium, and by the Public Health England project on invasive mosquito surveillance. The findings of our study demonstrate the aircraft mediated transport of EMS into Europe from a wide range of possible areas in the world. Results show accidental introductions of Aedes aegypti and Ae. albopictus, as well as exotic Anopheles and Mansonia specimens. The findings of Ae. albopictus at Schiphol airport are the first evidence of accidental introduction of the species using this pathway in Europe. Furthermore, our results stress the importance of the use of molecular tools to validate the morphology-based species identifications. We recommend monitoring of EMS at airports with special attention to locations with a high movement of cargo and passengers.
    Keywords:  DNA barcoding; disease vector; exotic mosquitoes; globalization; monitoring; public health; real-time PCR; species identification; temperate areas; vector surveillance
    DOI:  https://doi.org/10.3390/ijerph17103450
  11. Open Vet J. 2020 Apr;10(1): 39-43
    Zhang J, Lu G, Kelly P, Li J, Li M, Wang J, Huang K, Qiu H, You J, Zhang R, Wang Y, Zhang Y, Wang C.
      Background: Plasmodium relictum is one of the most important avian malaria species, which is mainly seen in wild birds, with infections reported in more than 70 different species and at high prevalence.Aim: The aim of this study was to determine the molecular prevalence of Plasmodium spp. in mosquitoes collected in China.
    Method: A Plasmodium -specific fluorescence resonance energy transfer (FRET) polymerase chain reaction (PCR) was established in this study to analyze five species of mosquitoes (1,620 Culex pipiens pallens, 806 Aedes albopictus, 377 Armigeres subalbatus, 168 Anopheles sinensis, and 80 Culex tritaeniorhynchus) collected in hand nets from homes in 25 provinces of China.
    Results: Only females originated from six provinces were determined to be positive (0.6%, 10/1,809). Plasmodium species were detected in three mosquito species, such as C. pipiens pallens (0.5%, 8/1,620), A. sinensis (0.6%, 1/168), and A. subalbatus (0.3%, 1/377). Of the three mosquito species positive for P. relictum, only C. pipiens pallens is known to feed on birds and is recognized as the natural vector of P. relictum.
    Conclusion: This is the first time that P. relictum has been detected in A. sinensis and A. subalbatus. P. relictum, the agent of avian malaria, was present in mosquitoes in China, including mosquito species not previously thought to be the vectors.
    Keywords:  Anopheles sinensis; Armigeres subalbatus; China; Mosquito; Plasmodium relictum
    DOI:  https://doi.org/10.4314/ovj.v10i1.7
  12. Acta Trop. 2020 May 13. pii: S0001-706X(19)31785-1. [Epub ahead of print] 105468
    Buchwald AG, Hayden MH, Dadzie SK, Paull SH, Carlton EJ.
      Arboviruses transmitted by Aedes mosquitoes are a growing global concern; however, there remain large gaps in surveillance of both arboviruses and their vectors in West Africa. We reviewed over 50 years of data including outbreak reports, peer-reviewed literature, and prior data compilations describing Zika, dengue, and chikungunya, and their vectors in West Africa. Large outbreaks of dengue, Zika, and chikungunya have recently occurred in the region with over 27,000 cases of Aedes-borne disease documented since 2007. Recent arboviral outbreaks have become more concentrated in urban areas, and Aedes albopictus, a relatively new arrival in the region, has emerged as an important vector in several areas. Seroprevalence surveys suggest reported cases are a gross underestimate of the underlying arboviral disease burden. These findings indicate a shifting epidemiology of arboviral disease in West Africa and highlight a need for increased research and implementation of vector and disease control. Rapid urbanization and climate change may further alter disease patterns, underscoring the need for improved diagnostic capacity, and vector and disease surveillance to address this evolving health challenge.
    Keywords:  Aedes aegypti; Aedes albopictus; Arboviruses; Climate change; Urbanization; West Africa
    DOI:  https://doi.org/10.1016/j.actatropica.2020.105468
  13. G3 (Bethesda). 2020 May 18. pii: g3.401279.2020. [Epub ahead of print]
    Chang X, Zhong D, Wang X, Bonizzoni M, Li Y, Zhou G, Cui L, Wei X, Yan G.
      Anopheles sinensis is a major malaria vector in Southeast Asia. Resistance to pyrethroid insecticides in this species has impeded malaria control in the region. Previous studies found that An. sinensis populations from Yunnan Province, China were highly resistant to deltamethrin and did not carry mutations in the voltage-gated sodium channel gene that cause knockdown resistance. In this study, we tested the hypothesis that other genomic variants are associated with the resistance phenotype. Using paired-end whole genome sequencing (DNA-seq), we generated 108 Gb of DNA sequence from deltamethrin -resistant and -susceptible mosquito pools with an average coverage of 83.3× depth. Using a stringent filtering method, we identified a total of 916,926 single nucleotide variants (SNVs), including 32,240 non-synonymous mutations. A total of 958 SNVs differed significantly in allele frequency between deltamethrin -resistant and -susceptible mosquitoes. Of these, 43 SNVs were present within 37 genes that code for immunity, detoxification, cuticular, and odorant proteins. A subset of 12 SNVs were randomly selected for genotyping of individual mosquitoes by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and showed consistent allele frequencies with the pooled DNA-seq derived allele frequencies. In addition, copy number variations (CNVs) were detected in 56 genes, including 33 that contained amplification alleles and 23 that contained deletion alleles in resistant mosquitoes compared to susceptible mosquitoes. The genomic variants described here provides a useful resource for future studies on the genetic mechanism of insecticide resistance in this important malaria vector species.
    Keywords:  Anopheles sinensis; copy number variation; genomic variant; insecticide resistance; polymerase chain reaction-restriction fragment length polymorphism; whole genome sequencing
    DOI:  https://doi.org/10.1534/g3.120.401279
  14. PLoS Negl Trop Dis. 2020 May 18. 14(5): e0007754
    Brackney DE, Correa MA, Cozens DW.
      Macroautophagy is an evolutionarily conserved cellular process critical for maintaining cellular homeostasis. It can additionally function as an innate immune response to viral infection as has been demonstrated for a number of arthropod-borne (arbo-) viruses. Arboviruses are maintained in a transmission cycle between vertebrate hosts and invertebrate vectors yet the majority of studies assessing autophagy-arbovirus interactions have been limited to the mammalian host. Therefore we evaluated the role of autophagy during arbovirus infection of the invertebrate vector using the tractable Aag2 Aedes aegypti mosquito cell culture system. Our data demonstrates that autophagy is significantly induced in mosquito cells upon infection with two divergent arboviruses: dengue virus-2 (DENV-2; Flaviviridae, Flavivirus) and chikungunya virus (CHIKV; Togaviridae, Alphavirus). While assessing the role of autophagy during arbovirus infection, we observed a somewhat paradoxical outcome. Both induction and suppression of autophagy via torin-1 and spautin-1, respectively, resulted in increased viral titers for both viruses, yet suppression of autophagy-related genes had no effect. Interestingly, chemical modulators of autophagy had either no effect or opposite effects in another widely used mosquito cell line, C6/36 Aedes albopictus cells. Together, our data reveals a limited role for autophagy during arbovirus infection of mosquito cells. Further, our findings suggest that commonly used chemical modulators of autophagy alter mosquito cells in such a way as to promote viral replication; however, it is unclear if this occurs directly through autophagic manipulation or other means.
    DOI:  https://doi.org/10.1371/journal.pntd.0007754
  15. Front Vet Sci. 2020 ;7 243
    Calzolari M, Angelini P, Bolzoni L, Bonilauri P, Cagarelli R, Canziani S, Cereda D, Cerioli MP, Chiari M, Galletti G, Moirano G, Tamba M, Torri D, Trogu T, Albieri A, Bellini R, Lelli D.
      With several human cases reported annually since 2008 and the unapparent risk of infection of blood donors, the West Nile virus (WNV) is emerging as an important health issue in Europe. Italy, as well as other European countries, experienced a recrudescence of the virus circulation in 2018, which led to an increased number of human cases. An integrated surveillance plan was activated in the Emilia-Romagna and Lombardy regions (Northern Italy) since 2008 in order to monitor the intensity and timing of WNV circulation. A fundamental part of this plan consists in entomological surveillance. In 2018, the surveillance plan made it possible to collect 385,293 mosquitoes in 163 stations in the two Regions. In total 269,147 Culex mosquitoes were grouped into 2,337 pools and tested for WNV, which was detected in 232 pools. Circulation started in the central part of the Emilia-Romagna region in the middle of June, about one month before the previous seasons. Circulation suddenly expanded to the rest of the region and reached the Lombardy region in the middle of July. WNV circulated more intensively in the eastern part of the surveyed area, as confirmed by the highest number of human cases. A relationship between the number of mosquitoes collected and the virus incidence emerged, but the data obtained highlighted that the probability of detecting the virus in a given site was less than expected with a higher number of collected mosquitoes. A significant relationship was observed between the temperature recorded one week before the sampling and the number of collected mosquitoes, as well as between the estimated number of WNV-positive mosquitoes and the temperature recorded two weeks before the sampling. The two weeks delay in the influence of temperature on the positive mosquitoes is in line with the time of the virus extrinsic incubation in the mosquito. This finding confirms that temperature is one of the principal drivers in WNV mosquito infection. The surveillance system demonstrated the ability to detect the virus circulation early, particularly in areas where circulation was more intense. This allowed evaluating the effect of mosquito abundance and weather factors on virus circulation.
    Keywords:  Culex pipiens; West Nile virus; infection rate; one-health; surveillance; temperature
    DOI:  https://doi.org/10.3389/fvets.2020.00243
  16. Parasit Vectors. 2020 May 20. 13(1): 266
    Guissou E, Poda S, de Sales Hien DF, Yerbanga SR, Da DF, Cohuet A, Fournet F, Roux O, Maiga H, Diabaté A, Gilles J, Bouyer J, Ouédraogo AG, Rayaissé JB, Lefèvre T, Dabiré KR.
      BACKGROUND: The sterile insect technique (SIT) is a vector control strategy relying on the mass release of sterile males into wild vector populations. Current sex separation techniques are not fully efficient and could lead to the release of a small proportion of females. It is therefore important to evaluate the effect of irradiation on the ability of released females to transmit pathogens. This study aimed to assess the effect of irradiation on the survival and competence of Anopheles arabiensis females for Plasmodium falciparum in laboratory conditions.METHODS: Pupae were irradiated at 95 Gy of gamma-rays, and emerging females were challenged with one of 14 natural isolates of P. falciparum. Seven days post-blood meal (dpbm), irradiated and unirradiated-control females were dissected to assess the presence of oocysts, using 8 parasite isolates. On 14 dpbm, sporozoite dissemination in the head/thorax was also examined, using 10 parasites isolates including 4 in common with the 7 dpbm dissection (oocyst data). The survivorship of irradiated and unirradiated-control mosquitoes was monitored.
    RESULTS: Overall, irradiation reduced the proportion of mosquitoes infected with the oocyst stages by 17% but this effect was highly inconsistent among parasite isolates. Secondly, there was no significant effect of irradiation on the number of developing oocysts. Thirdly, there was no significant difference in both the sporozoite infection rate and load between the irradiated and unirradiated-control mosquitoes. Fourthly, irradiation had varying effects on female survival with either a negative effect or no effect.
    CONCLUSIONS: The effect of irradiation on mosquito competence strongly varied among parasite isolates. Because of such isolate variability and, the fact that different parasite isolates were used to collect oocyst and sporozoite data, the irradiation-mediated reduction of oocyst prevalence was not confirmed for the sporozoite stages. Our data indicate that irradiated female An. arabiensis could contribute to malaria transmission, and highlight the need for perfect sexing tools, which would prevent the release of females as part of SIT programmes.
    Keywords:  Competence; Direct membrane feeding assay; Sterile insect technique (SIT)
    DOI:  https://doi.org/10.1186/s13071-020-04135-w
  17. J Virol Methods. 2020 May 14. pii: S0166-0934(20)30114-2. [Epub ahead of print] 113862
    Ou TP, Yun C, Auerswald H, In S, Leang R, Huy R, Chhoeung R, Dussart P, Duong V.
      Dengue virus (DENV) and Zika virus (ZIKV) are important viral pathogens, known to cause human infections with similar symptoms, are transmitted by common vectors and co-circulate in intertropical regions. Moreover, dengue fever results from infection with one of four different serotypes of dengue virus. Considering the recent ZIKV emergence, multiplex and up-to-date assays are more preferable for detection of both viruses in a single reaction. This study aimed to develop: (i) an one-step duplex real-time reverse transcription polymerase chain reaction (RT-qPCR) assay to efficiently and simultaneously detect and quantify DENV and ZIKV; (ii) a fourplex RT-qPCR to differentiate and quantify the four DENV serotypes. The detection limit of the duplex assay was 0.028 and 0.065 FFU (focus forming unit)/ml for DENV and ZIKV respectively. The lower limit of analytical sensitivity of fourplex assay was 0.01 FFU/ml for DENV-1 and 0.1 FFU/ml for DENV-2,-3 and -4. The assessment of specificity indicated both assays were highly specific to targeted viruses with negative results for other Flaviviridae such as Japanese encephalitis, West Nile, Yellow fever or Hepatitis C viruses. The newly developed RT-qPCRs were shown to be more sensitive than a previously described assay in detecting DENV in clinical samples and are suitable for the routine diagnosis.
    Keywords:  Zika virus; arbovirus; dengue virus; diagnosis; fourplex real-time RT-qPCR; one-step duplex real-time RT-qPCR
    DOI:  https://doi.org/10.1016/j.jviromet.2020.113862
  18. Viruses. 2020 May 18. pii: E558. [Epub ahead of print]12(5):
    Mapder T, Aaskov J, Burrage K.
      The host-vector shuttle and the bottleneck in dengue transmission is a significant aspect with regard to the study of dengue outbreaks. As mosquitoes require 100-1000 times more virus to become infected than human, the transmission of dengue virus from human to mosquito is a vulnerability that can be targeted to improve disease control. In order to capture the heterogeneity in the infectiousness of an infected patient population towards the mosquito population, we calibrate a population of host-to-vector virus transmission models based on an experimentally quantified infected fraction of a mosquito population. Once the population of models is well-calibrated, we deploy a population of controls that helps to inhibit the human-to-mosquito transmission of the dengue virus indirectly by reducing the viral load in the patient body fluid. We use an optimal bang-bang control on the administration of the defective virus (transmissible interfering particles (TIPs)) to symptomatic patients in the course of their febrile period and observe the dynamics in successful reduction of dengue spread into mosquitoes.
    Keywords:  defective particles; dengue transmission; dengue virus; mosquito infectious dose; population of models
    DOI:  https://doi.org/10.3390/v12050558
  19. Viruses. 2020 May 19. pii: E561. [Epub ahead of print]12(5):
    Holicki CM, Ziegler U, Răileanu C, Kampen H, Werner D, Schulz J, Silaghi C, Groschup MH, Vasić A.
      West Nile virus (WNV) is a widespread zoonotic arbovirus and a threat to public health in Germany since its first emergence in 2018. It has become of particular relevance in Germany in 2019 due to its rapid geographical spread and the detection of the first human clinical cases. The susceptibility of indigenous Culex pipiens (biotypes pipiens and molestus) for a German WNV lineage 2 strain was experimentally compared to that of Serbian Cx. pipiens biotype molestus and invasive German Aedes albopictus. All tested populations proved to be competent laboratory vectors of WNV. Culex pipiens biotype pipiens displayed the highest transmission efficiencies (40.0%-52.9%) at 25 °C. This biotype was also able to transmit WNV at 18 °C (transmission efficiencies of 4.4%-8.3%), proving that temperate climates in Central and Northern Europe may support WNV circulation. Furthermore, due to their feeding behaviors, Cx. pipiens biotype molestus and Ae. albopictus can act as "bridge vectors", leading to human WNV infections.
    Keywords:  Aedes albopictus; Culex pipiens; West Nile virus; arbovirus; transmission; vector competence
    DOI:  https://doi.org/10.3390/v12050561
  20. Am J Trop Med Hyg. 2020 May 18.
    Robison A, Young MC, Byas AD, Rückert C, Ebel GD.
      Chikungunya virus (CHIKV) and Zika virus (ZIKV) are arthropod-borne viruses transmitted mainly by Aedes aegypti mosquitoes. These viruses have become endemic in large parts of North, Central, and South America. Arboviruses persistently infect mosquitoes throughout their life span and become infectious (i.e., expectorate infectious virus in saliva) after a period of time called the extrinsic incubation period (EIP). The duration of this infectiousness, however, is not well characterized. This is an important shortcoming because many epidemiological models assume that mosquitoes continue to be infectious for the duration of their life span. To define the duration of infectiousness for CHIKV and ZIKV, mosquitoes were infected orally with these viruses. Every 2 days, legs/wings, midguts, salivary glands, and saliva were collected from 30 to 60 mosquitoes and viral load measured. In CHIKV-infected mosquitoes, infectious virus in saliva peaked early (2-4 dpi), and then decreased rapidly and was rarely observed after 10 dpi. Viral RNA in infected tissues also decreased after the initial peak (4-8 dpi) but did so much less drastically. In ZIKV-infected mosquitoes, the infectious virus in saliva peaked at 12-14 dpi and dropped off only slightly after 14 dpi. In infected tissues, viral RNA increased early during infection, and then plateaued after 6-10 days. Our findings suggest that significant variation exists in the duration of the infectious period for arboviruses that is in part influenced by virus clearance from expectorated saliva.
    DOI:  https://doi.org/10.4269/ajtmh.20-0143
  21. Curr Opin Insect Sci. 2020 Apr 04. pii: S2214-5745(20)30045-6. [Epub ahead of print]40 6-10
    Ignell R, Hill SR.
      The field of mosquito chemical ecology has shifted focus over the past five years, driven by the recognition that odour-mediated behaviours are regulated by distinct chemical codes, that is, odour blends emanating from the natural environment. As a research community, we have shifted from our anthropocentric focus to include other behaviours, including plant seeking and oviposition site seeking, in order to develop new tools to combat residual malaria in the wake of the increased insecticide and behavioural resistance in mosquitoes across sub-Saharan Africa. In this short review, we will outline the progress made, and the future directions, in understanding blend recognition and chemical parsimony, and their implications for preadaptation of the odour coding system in malaria mosquitoes.
    DOI:  https://doi.org/10.1016/j.cois.2020.03.008
  22. Global Health. 2020 May 18. 16(1): 47
    Riccardo F, Bolici F, Fafangel M, Jovanovic V, Socan M, Klepac P, Plavsa D, Vasic M, Bella A, Diana G, Rosi L, Pezzotti P, Andrianou XD, Di Luca M, Venturi G, Maraglino F, Pervanidou D, Cenciarelli O, Baka A, Young J, Bakonyi T, Rezza G, Suk JE.
      BACKGROUND: After Action Reviews (AAR) with a One Health perspective were performed in Slovenia, Italy, Serbia and Greece following a severe West Nile virus (WNV) transmission season in 2018. A protocol combining traditional techniques and organizational process analysis was developed and then implemented in each country.RESULTS: In 2018, response to the unusually intense transmission season of WNV in Slovenia, Italy, Serbia and Greece took place through routine response mechanisms. None of the four countries declared a national or subnational emergency. We found a very strong consensus on the strengths identified in responding to this event. All countries indicated the availability of One Health Plans for surveillance and response; very high laboratory diagnostic capacity in the human, veterinary and entomology sectors and strong inter-sectoral collaboration with strong commitment of engaged institutions as critical in the management of the event. Finally, countries implementing One Health surveillance for WNV (in terms of early warning and early activation of prevention measures) consistently reported a positive impact on their activities, in particular when combining mosquito and bird surveillance with surveillance of cases in humans and equids. Recurring priority areas for improvement included: increasing knowledge on vector-control measures, ensuring the sustainability of vector monitoring and surveillance, and improving capacity to manage media pressure.
    CONCLUSIONS: The AARs presented here demonstrate the benefit of cross-sectoral and cross-disciplinary approaches to preparedness for West Nile virus outbreaks in Europe. In the coming years, priorities include fostering and strengthening arrangements that: enable coordinated One Health surveillance and response during WNV transmission seasons; ensure adequate laboratory capacities; strengthen risk communication; and fund longer-term research to address the knowledge gaps identified in this study.
    Keywords:  After action reviews; Epidemics; Infectious disease; Mosquito-borne disease; Outbreaks; Preparedness; West Nile virus
    DOI:  https://doi.org/10.1186/s12992-020-00568-1
  23. Sci Adv. 2020 May;6(20): eaay5487
    Brydegaard M, Jansson S, Malmqvist E, Mlacha YP, Gebru A, Okumu F, Killeen GF, Kirkeby C.
      Yearly, a quarter billion people are infected and a half a million killed by the mosquito-borne disease malaria. Lack of real-time observational tools for continuously assessing the unperturbed mosquito flight activity in situ limits progress toward improved vector control. We deployed a high-resolution entomological lidar to monitor a half-kilometer static transect adjacent to a Tanzanian village. We evaluated one-third million insect observations during five nights, four days, and one annular solar eclipse. We demonstrate in situ lidar classification of several insect families and their sexes based on their modulation signatures. We were able to compare the fine-scale spatiotemporal activity patterns of malaria vectors during ordinary days and an eclipse to disentangle phototactic activity patterns from the circadian mechanism. We observed an increased insect activity during the eclipse attributable to mosquitoes. These unprecedented findings demonstrate how lidar-based monitoring of distinct mosquito activities could advance our understanding of vector ecology.
    DOI:  https://doi.org/10.1126/sciadv.aay5487
  24. Am J Trop Med Hyg. 2020 May 18.
    Syafruddin D, Asih PBS, Rozi IE, Permana DH, Nur Hidayati AP, Syahrani L, Zubaidah S, Sidik D, Bangs MJ, Bøgh C, Liu F, Eugenio EC, Hendrickson J, Burton T, Baird KJ, Collins F, Grieco JP, Lobo NF, Achee NL.
      A cluster-randomized, double-blinded, placebo-controlled trial was conducted to estimate the protective efficacy (PE) of a spatial repellent (SR) against malaria infection in Sumba, Indonesia. Following radical cure in 1,341 children aged ≥ 6 months to ≤ 5 years in 24 clusters, households were given transfluthrin or placebo passive emanators (devices designed to release vaporized chemical). Monthly blood screening and biweekly human-landing mosquito catches were performed during a 10-month baseline (June 2015-March 2016) and a 24-month intervention period (April 2016-April 2018). Screening detected 164 first-time infections and an accumulative total of 459 infections in 667 subjects in placebo-control households, and 134 first-time and 253 accumulative total infections among 665 subjects in active intervention households. The 24-cluster protective effect of 27.7% and 31.3%, for time to first-event and overall (total new) infections, respectively, was not statistically significant. Purportedly, this was due in part to zero to low incidence in some clusters, undermining the ability to detect a protective effect. Subgroup analysis of 19 clusters where at least one infection occurred during baseline showed 33.3% (P-value = 0.083) and 40.9% (P-value = 0.0236, statistically significant at the one-sided 5% significance level) protective effect to first infection and overall infections, respectively. Among 12 moderate- to high-risk clusters, a statistically significant decrease in infection by intervention was detected (60% PE). Primary entomological analysis of impact was inconclusive. Although this study suggests SRs prevent malaria, additional evidence is required to demonstrate the product class provides an operationally feasible and effective means of reducing malaria transmission.
    DOI:  https://doi.org/10.4269/ajtmh.19-0554
  25. Acta Trop. 2020 May 15. pii: S0001-706X(20)30297-7. [Epub ahead of print]208 105514
    Papa A, Tsioka K, Gewehr S, Kalaitzopoulou S, Pappa S, Mourelatos S.
      West Nile virus is a flavivirus transmitted to humans mainly by mosquito bites. Outbreaks are observed in several European countries, and Greece is one of the most affected countries during the recent years. Thessaly was one of the most affected regions in Greece in 2019. A total of 3,025 Culex spp. mosquitoes collected in Thessaly were grouped into 47 pools and tested for West Nile virus (WNV). Eight (17%) pools were found positive. Whole genome sequences were obtained from two positive pools. Phylogenetic analysis showed that the causative strain was an evolutionary variant of the strains circulating in 2018 belonging to the Balkan subgroup of the Central European subclade of WNV lineage 2.
    Keywords:  Culex spp.; Lineage 2; Mosquitoes; West Nile virus; Whole genome
    DOI:  https://doi.org/10.1016/j.actatropica.2020.105514
  26. Sci Adv. 2020 May;6(20): eaay5898
    Dong Y, Simões ML, Dimopoulos G.
      The malaria parasite's complex journey through the Anopheles mosquito vector provides multiple opportunities for targeting Plasmodium with recombinant effectors at different developmental stages and different host tissues. We have designed and expressed transgenes that efficiently suppress Plasmodium infection by targeting the parasite with multiple independent endogenous and exogenous effectors at multiple infection stages to potentiate suppression and minimize the probability for development of resistance to develop. We have also addressed the fitness impact of transgene expression on the mosquito. We show that highly potent suppression can be achieved by targeting both pre-oocyst stages by transgenically overexpressing either the endogenous immune deficiency immune pathway transcription factor Rel2 or a polycistronic mRNA encoding multiple antiparasitic effectors and simultaneously targeting the sporozoite stages with an anti-sporozoite single-chain antibody fused to the antiparasitic protein Scorpine. Expression of the selected endogenous effector systems appears to pose a lower fitness cost than does the use of foreign genes.
    DOI:  https://doi.org/10.1126/sciadv.aay5898
  27. PLoS Negl Trop Dis. 2020 May 19. 14(5): e0008300
    Zhou CM, Liu JW, Qi R, Fang LZ, Qin XR, Han HJ, Mo RC, Yu H, Jiao YJ, Lin JY, Yu XJ.
      Currently, Zika virus (ZIKV) is spreading across the world and no ZIKV infection cases have ever been reported in China. Here, we aimed to determine whether ZIKV infection exists in China. Blood samples of 273 healthy individuals were collected from Nanning City, Guangxi Province, China in March 2019. We found that 9.5% (26/273) and 1.8% (5/273) of healthy persons were positive to ZIKV total antibody (IgG and/or IgM) IgM antibody, respectively. All ZIKV positive plasma samples were negative to Dengue virus and West Nile virus. Among the ZIKV antibody positive plasma samples, 65.4% (17/26) exhibited neutralizing activity to ZIKV. Followed up studies showed that none had clinical symptoms of ZIKV infection and oversea experience. Together, our study indicates that endemic ZIKV infections emerge in China, which not only suggested that ZIKV posed a potential threat to public health in China, but also expand the ZIKV epidemic areas in East and Southeast Asia.
    DOI:  https://doi.org/10.1371/journal.pntd.0008300
  28. J Biol Chem. 2020 May 22. pii: jbc.RA120.013869. [Epub ahead of print]
    Onofre J, Pacheco S, Torres-Quintero MC, Gill SS, Soberon M, Bravo A.
      Bacillus thuringiensis subsp. israelensis (Bti) produces crystal inclusions composed of three-domain Cry proteins and cytolytic Cyt toxins, which are toxic to different mosquito larvae. A key component is the Cyt toxin, which synergizes the activity of the other Cry toxins, thereby resulting in high toxicity. The precise mechanism of action of Cyt toxins is still debated, and two models have been proposed, the pore formation model and the detergent effect. Here, we performed a systematic structural characterization of the Cyt toxin interaction with different membranes, including in Aedes aegypti larval brush border membrane vesicles (BBMVs), small unilamellar vesicles (SUVs) liposomes, and rabbit erythrocytes. We examined Cyt1Aa insertion into these membranes by analyzing fluorescence quenching in solution and in the membrane-bound state. For this purpose, we constructed several Cyt1Aa variants having substitutions with a single cysteine residue in different secondary structures, enabling Cys labeling with Alexa Fluor-488 for quenching analysis using I-soluble quencher in solution and in the membrane-bound state. We identified the Cyt1Aa residues exposed to the solvent upon membrane insertion, predicting a possible topology of the membrane-inserted toxin in the different membranes. Moreover, toxicity assays with these variants revealed that Cyt1Aa exerts its insecticidal activity and hemolysis through different mechanisms. We found that Cyt1Aa exhibits variable interactions with each membrane system, with deeper insertion into mosquito larva membranes, supporting the pore formation model, whereas in the case of erythrocytes and SUVs, Cyt1Aa's insertion was more superficial, supporting the notion that a detergent effect underlies its hemolytic activity.
    Keywords:  Bacillus; Bacillus thuringiensis; Fluorescence quenching; bacterial toxin; conformational change; fluorescence; membrane; membrane insertion; pore formation; toxin
    DOI:  https://doi.org/10.1074/jbc.RA120.013869
  29. Pest Manag Sci. 2020 May 20.
    Puig-Montserrat X, Flaquer C, Gómez-Aguilera N, Burgas A, Mas M, Tuneu C, Marquès E, López-Baucells A.
      BACKGROUND: The fact that bats suppress agricultural pests has been measured for some particular dyads of predator and prey species both in economic and food security terms. The recent emergence of new molecular techniques allows for wide screenings of bat diet and provide a further evidence that bats consume an ample array of agricultural pest species. The main focus about the regulatory services that bats provide in agroecosystems has been on crop pests that cause yield losses. Rice paddies constitute a particular agronomic system with specific challenges, not only related to crop productivity but also to human health. Dipteran density in such ecosystems poses a serious threat to human wellbeing and hinders crop production. Mosquitoes cause direct harm to human populations transmitting a number of infectious diseases. Non-biting midges (Chironomidae) can consume and weaken rice seedlings and can cause major yield losses.RESULTS: Mosquito populations and bat activity were assessed in rice paddies of Montgrí, Medes i Baix Ter Natural Park (NE Iberian Peninsula). Molecular analyses of bats faeces (6 weekly samples of 15 faeces each between mid-August and September) proved the presence of both mosquitoes and non-biting midges in all of the diet samples. Furthermore, bat activity at the sampling locations was related to adult mosquito density.
    CONCLUSION: Our results suggest that bats actively exploit the emergence of adult mosquitoes and further prove that they prey on mosquitoes, non-biting midges and other deleterious insects. Promoting the presence of bats next to human settlements in such agroecosystems may constitute a biological control system with direct impact on both human health and crop yield. This article is protected by copyright. All rights reserved.
    Keywords:  Chironomidae; Chiroptera; Culicidae; DNA metabarcoding; bats; biological pest control; mosquito-borne disease; rice
    DOI:  https://doi.org/10.1002/ps.5925
  30. Mem Inst Oswaldo Cruz. 2020 ;pii: S0074-02762020000100320. [Epub ahead of print]115 e190437
    Barbosa RMR, Melo-Santos MAV, Silveira JC, Silva-Filha MHNL, Souza WV, Oliveira CMF, Ayres CFJ, Xavier MDN, Rodrigues MP, Santos SAD, Nakazawa MM, Regis LN.
      BACKGROUND Aedes aegypti and Aedes albopictus are the most important arbovirus vectors in the world. OBJECTIVES This study aimed to investigate and compare the infestation pattern of these species in a neighbourhood of Recife, Brazil, endemic for arboviruses in 2005 (T1) and 2013 (T2). METHODS Infestation, distribution and relative abundance of these sympatric species were recorded by egg collection using a network of 59 sentinel ovitraps (s-ovt) at fixed sampling stations for 12 months in T1 and T2. FINDINGS A permanent occupation pattern was detected which was characterised by the presence of egg-laying females of one or both species with a high ovitrap positivity index (94.3 to 100%) throughout both years analysed. In terms of abundance, the total of eggs collected was lower (p < 0.005) in T2 (146,153) than in T1 (281,103), although ovitraps still displayed a high index of positivity. The spatial distribution showed the presence of both species in 65.1% of the 148 s-ovt assessed, while a smaller number of traps exclusively contained Ae. aegypti (22%) or Ae. albopictus (13.2%) eggs. MAIN CONCLUSIONS Our comparative analysis demonstrated the robustness of the spatial occupation and permanence of Ae. aegypti and Ae. albopictus populations in this endemic urban area.
    DOI:  https://doi.org/10.1590/0074-02760190437