Vectors and Vector-borne Diseases

A special issue of Insects (ISSN 2075-4450). This special issue belongs to the section "Insect Pest and Vector Management".

Deadline for manuscript submissions: closed (30 September 2019) | Viewed by 58373

Special Issue Editors


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Guest Editor
Department of Biological Sciences, Texas Tech University, Box 43131, Lubbock, TX 79409, USA
Interests: vector control; Wolbachia; vector-borne disease

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Guest Editor
Assistant Professor, Department of Biochemistry and Molecular Biology, University of Nevada, Reno, NV 89557, USA
Interests: arthropod genomics; arthropod physiology; vector biology; vector-pathogen interactions

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Guest Editor
Baylor University, Waco, TX, USA
Interests: sensory biology; urban medical entomology; chemical ecology

Special Issue Information

Dear Colleagues,

Vector biology is the study of arthropod vectors and their role in pathogen transmission. Despite significant advances in the study of vector biology, vector-borne diseases still account for more than 17% of infectious diseases. Unfortunately, these advances and the contribution of the vector to the process of pathogen transmission is often overlooked, regardless of the potential for new methods to combat vector-borne diseases. For this Special Issue, we encourage authors to contribute original research articles on the current status of arthropod vectors and vector-borne diseases, with a special emphasis on alternative approaches to controlling vectors of disease, host–pathogen–symbiont interactions, insect pathogen resistance, functional genomics, vector ecology, and vector competence.

Prof. Corey Brelsfoard
Prof. Monika Gulia-Nuss
Prof. Jason Pitts
Guest Editors

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Keywords

  • Vector biology
  • host pathogen interactions
  • vector competence
  • vector ecology

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Published Papers (12 papers)

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Research

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14 pages, 1909 KiB  
Article
Blood Digestion by Trypsin-Like Serine Proteases in the Replete Lyme Disease Vector Tick, Ixodes scapularis
by Jeremiah Reyes, Cuauhtemoc Ayala-Chavez, Arvind Sharma, Michael Pham, Andrew B. Nuss and Monika Gulia-Nuss
Insects 2020, 11(3), 201; https://doi.org/10.3390/insects11030201 - 23 Mar 2020
Cited by 18 | Viewed by 6506
Abstract
Ixodes scapularis is the major vector of Lyme disease in the Eastern United States. Each active life stage (larva, nymph, and adult) takes a blood meal either for developing and molting to the next stage (larvae and nymphs) or for oviposition (adult females). [...] Read more.
Ixodes scapularis is the major vector of Lyme disease in the Eastern United States. Each active life stage (larva, nymph, and adult) takes a blood meal either for developing and molting to the next stage (larvae and nymphs) or for oviposition (adult females). This protein-rich blood meal is the only food taken by Ixodes ticks and therefore efficient blood digestion is critical for survival. Studies in partially engorged ticks have shown that the initial stages of digestion are carried out by cathepsin proteases within acidic digestive cells. In this study, we investigated the potential role of serine proteases in blood digestion in replete ticks. RNA interference was used for functional analysis and a trypsin-benzoyl-D, L-arginine 4-nitoanilide assay was used to measure active trypsin levels. Hemoglobinolytic activity was determined in vitro, with or without a serine protease inhibitor. Our data suggest that trypsin levels increase significantly after repletion. Knockdown of serine proteases negatively impacted blood feeding, survival, fecundity, levels of active trypsin in the midgut, and resulted in lower hemoglobin degradation. Incubation of midgut extract with a trypsin inhibitor resulted in 65% lower hemoglobin degradation. We provide evidence of the serine proteases as digestive enzymes in fully engorged, replete females. Understanding the digestive profile of trypsin during blood meal digestion in I. scapularis improves our understanding of the basic biology of ticks and may lead to new methods for tick control. Full article
(This article belongs to the Special Issue Vectors and Vector-borne Diseases)
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15 pages, 4252 KiB  
Article
The Influence of New Surveillance Data on Predictive Species Distribution Modeling of Aedes aegypti and Aedes albopictus in the United States
by Hannah S. Tiffin, Steven T. Peper, Alexander N. Wilson-Fallon, Katelyn M. Haydett, Guofeng Cao and Steven M. Presley
Insects 2019, 10(11), 400; https://doi.org/10.3390/insects10110400 - 12 Nov 2019
Cited by 8 | Viewed by 3227
Abstract
The recent emergence or reemergence of various vector-borne diseases makes the knowledge of disease vectors’ presence and distribution of paramount concern for protecting national human and animal health. While several studies have modeled Aedes aegypti or Aedes albopictus distributions in the past five [...] Read more.
The recent emergence or reemergence of various vector-borne diseases makes the knowledge of disease vectors’ presence and distribution of paramount concern for protecting national human and animal health. While several studies have modeled Aedes aegypti or Aedes albopictus distributions in the past five years, studies at a large scale can miss the complexities that contribute to a species’ distribution. Many localities in the United States have lacked or had sporadic surveillance conducted for these two species. To address these gaps in the current knowledge of Ae. aegypti and Ae. albopictus distributions in the United States, surveillance was focused on areas in Texas at the margins of their known ranges and in localities that had little or no surveillance conducted in the past. This information was used with a global database of occurrence records to create a predictive model of these two species’ distributions in the United States. Additionally, the surveillance data from Texas was used to determine the influence of new data from the margins of a species’ known range on predicted species’ suitability maps. This information is critical in determining where to focus resources for the future and continued surveillance for these two species of medical concern. Full article
(This article belongs to the Special Issue Vectors and Vector-borne Diseases)
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16 pages, 2132 KiB  
Article
Insecticidal and Antifeedant Activities of Malagasy Medicinal Plant (Cinnamosma sp.) Extracts and Drimane-Type Sesquiterpenes against Aedes aegypti Mosquitoes
by Edna Alfaro Inocente, Bao Nguyen, Preston K. Manwill, Annecie Benatrehina, Eliningaya Kweka, Sijin Wu, Xiaolin Cheng, L. Harinantenaina Rakotondraibe and Peter M. Piermarini
Insects 2019, 10(11), 373; https://doi.org/10.3390/insects10110373 - 25 Oct 2019
Cited by 18 | Viewed by 5550
Abstract
The overuse of insecticides with limited modes of action has led to resistance in mosquito vectors. Thus, insecticides with novel modes of action are needed. Secondary metabolites in Madagascan plants of the genus Cinnamosma (Canellaceae) are commonly used in traditional remedies and known [...] Read more.
The overuse of insecticides with limited modes of action has led to resistance in mosquito vectors. Thus, insecticides with novel modes of action are needed. Secondary metabolites in Madagascan plants of the genus Cinnamosma (Canellaceae) are commonly used in traditional remedies and known to elicit antifeedant and toxic effects in insect pests. Here we test the hypothesis that extracts of Cinnamosma sp. enriched in drimane sesquiterpenes are toxic and/or antifeedant to the yellow fever mosquito Aedes aegypti. We show that the bark and root extracts, which contain a higher abundance of drimane sesquiterpenes compared to leaves, were the most efficacious. Screening isolated compounds revealed cinnamodial to be the primary driver of adulticidal activity, whereas cinnamodial, polygodial, cinnafragrin A, and capsicodendrin contributed to the larvicidal activity. Moreover, an abundant lactone (cinnamosmolide) in the root extract synergized the larvicidal effects of cinnamodial. The antifeedant activity of the extracts was primarily contributed to cinnamodial, polygodial, and cinnamolide. Parallel experiments with warburganal isolated from Warburgia ugandensis (Canellaceae) revealed that aldehydes are critical for—and a hydroxyl modulates—insecticidal activity. Our results indicate that plant drimane sesquiterpenes provide valuable chemical platforms for developing insecticides and repellents to control mosquito vectors. Full article
(This article belongs to the Special Issue Vectors and Vector-borne Diseases)
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12 pages, 1372 KiB  
Article
Comparative Microbiome Profiles of Sympatric Tick Species from the Far-Western United States
by Betsabel Chicana, Lisa I. Couper, Jessica Y. Kwan, Enxhi Tahiraj and Andrea Swei
Insects 2019, 10(10), 353; https://doi.org/10.3390/insects10100353 - 18 Oct 2019
Cited by 43 | Viewed by 4919
Abstract
Insight into the composition and function of the tick microbiome has expanded considerably in recent years. Thus far, tick microbiome studies have focused on species and life stages that are responsible for transmitting disease. In this study we conducted extensive field sampling of [...] Read more.
Insight into the composition and function of the tick microbiome has expanded considerably in recent years. Thus far, tick microbiome studies have focused on species and life stages that are responsible for transmitting disease. In this study we conducted extensive field sampling of six tick species in the far-western United States to comparatively examine the microbial composition of sympatric tick species: Ixodes pacificus, Ixodes angustus, Dermacentor variabilis, Dermacentor occidentalis, Dermacentor albipictus, and Haemaphysalis leporispalustris. These species represent both common vectors of disease and species that rarely encounter humans, exhibiting a range of host preferences and natural history. We found significant differences in microbial species diversity and composition by tick species and life stage. The microbiome of most species examined were dominated by a few primary endosymbionts. Across all species, the relative abundance of these endosymbionts increased with life stage while species richness and diversity decreased with development. Only one species, I. angustus, did not show the presence of a single dominant microbial species indicating the unique physiology of this species or its interaction with the surrounding environment. Tick species that specialize in a small number of host species or habitat ranges exhibited lower microbiome diversity, suggesting that exposure to environmental conditions or host blood meal diversity can affect the tick microbiome which in turn may affect pathogen transmission. These findings reveal important associations between ticks and their microbial community and improve our understanding of the function of non-pathogenic microbiomes in tick physiology and pathogen transmission. Full article
(This article belongs to the Special Issue Vectors and Vector-borne Diseases)
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13 pages, 1782 KiB  
Article
Temperature and Sugar Feeding Effects on the Activity of a Laboratory Strain of Aedes aegypti
by Irvin Forde Upshur, Elizabeth Annadel Bose, Cameron Hart and Chloé Lahondère
Insects 2019, 10(10), 347; https://doi.org/10.3390/insects10100347 - 16 Oct 2019
Cited by 12 | Viewed by 5937
Abstract
Aedes aegypti is an invasive mosquito species that is expected to expand its global distribution through climate change. As poikilotherms, mosquitoes are greatly affected by the temperature of the environment which can impact host-seeking, blood-feeding, and flight activity as well as survival and [...] Read more.
Aedes aegypti is an invasive mosquito species that is expected to expand its global distribution through climate change. As poikilotherms, mosquitoes are greatly affected by the temperature of the environment which can impact host-seeking, blood-feeding, and flight activity as well as survival and ability to transmit pathogens. However, an important aspect of mosquito biology on which the effect of temperature has not been investigated is water and sugar-feeding and how access to a sugar source might affect the insect’s activity and survival under different thermal conditions. To close this knowledge gap, we relied on actometer experiments to study the activity of both female and male Ae. aegypti at 20 °C, 25 °C, and 30 °C, providing either water or 10% sucrose to the insects. We then measured the total carbohydrate contents of alive mosquitoes using the anthrone protocol. Survival was assessed and compared between all groups. Results from this study will inform on the thermal biology of Ae. aegypti mosquitoes and how access to sugar affects their activity. Full article
(This article belongs to the Special Issue Vectors and Vector-borne Diseases)
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9 pages, 1812 KiB  
Article
Inhibitors of Eicosanoid Biosynthesis Reveal that Multiple Lipid Signaling Pathways Influence Malaria Parasite Survival in Anopheles gambiae
by Hyeogsun Kwon and Ryan C. Smith
Insects 2019, 10(10), 307; https://doi.org/10.3390/insects10100307 - 20 Sep 2019
Cited by 5 | Viewed by 3295
Abstract
Eicosanoids are bioactive signaling lipids derived from the oxidation of fatty acids that act as important regulators of immune homeostasis and inflammation. As a result, effective anti-inflammatory drugs have been widely used to reduce pain and inflammation which target key eicosanoid biosynthesis enzymes. [...] Read more.
Eicosanoids are bioactive signaling lipids derived from the oxidation of fatty acids that act as important regulators of immune homeostasis and inflammation. As a result, effective anti-inflammatory drugs have been widely used to reduce pain and inflammation which target key eicosanoid biosynthesis enzymes. Conserved from vertebrates to insects, the use of these eicosanoid pathway inhibitors offer opportunities to evaluate the roles of eicosanoids in less-characterized insect systems. In this study, we examine the potential roles of eicosanoids on malaria parasite survival in the mosquito Anopheles gambiae. Using Plasmodium oocyst numbers to evaluate parasite infection, general or specific inhibitors of eicosanoid biosynthesis pathways were evaluated. Following the administration of dexamethasone and indomethacin, respective inhibitors of phospholipid A2 (PLA2) and cyclooxygenase (COX), oocyst numbers were unaffected. However, inhibition of lipoxygenase (LOX) activity through the use of esculetin significantly increased oocyst survival. In contrast, 12-[[(tricyclo[3.3.1.13,7]dec-1-ylamino)carbonyl]amino]-dodecanoic acid (AUDA), an inhibitor of epoxide hydroxylase (EH), decreased oocyst numbers. These experiments were further validated through RNAi experiments to silence candidate genes homologous to EH in An. gambiae to confirm their contributions to Plasmodium development. Similar to the results of AUDA treatment, the silencing of EH significantly reduced oocyst numbers. These results imply that specific eicosanoids in An. gambiae can have either agonist or antagonistic roles on malaria parasite survival in the mosquito host. Full article
(This article belongs to the Special Issue Vectors and Vector-borne Diseases)
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13 pages, 15672 KiB  
Article
Host Associations of Culex (Melanoconion) atratus (Diptera: Culicidae) and Culex (Melanoconion) pilosus from Florida, USA
by Lawrence E. Reeves, Isaiah Hoyer, Carolina Acevedo and Nathan D. Burkett-Cadena
Insects 2019, 10(8), 239; https://doi.org/10.3390/insects10080239 - 3 Aug 2019
Cited by 13 | Viewed by 4735
Abstract
Characterizing the host-use patterns of mosquitoes is an essential component of understanding the transmission dynamics of mosquito-vectored pathogens. The host associations of two species of the medically important Culex subgenus Melanoconion, Culex atratus, and Culex pilosus are unknown or unclear, respectively. Both [...] Read more.
Characterizing the host-use patterns of mosquitoes is an essential component of understanding the transmission dynamics of mosquito-vectored pathogens. The host associations of two species of the medically important Culex subgenus Melanoconion, Culex atratus, and Culex pilosus are unknown or unclear, respectively. Both species have wide neotropical distributions. In the United States of America (USA), Culex pilosus occurs throughout the southeastern coastal plain, while Culex atratus is restricted to the southern Florida Peninsula. Using PCR-based blood meal analysis, we investigated the host associations of Culex atratus and Culex pilosus that were collected from Everglades National Park, Florida, USA We identified the host species of 208 Culex atratus and 168 Culex pilosus. Both species were narrowly associated with reptilian host species, particularly native and non-native lizards of the genus Anolis. Sampled Culex atratus exclusively fed on reptilian hosts, with >99% of blood meals derived from Anolis lizards. Culex pilosus fed predominantly from reptiles, but avian and mammalian blood meals were also detected. Of these, 92% of blood meals were derived from Anolis species. For both species, Anolis sagrei, an invasive exotic lizard in Florida, was the most frequently detected host species. These data indicate that Culex atratus and Culex pilosus are specialists of reptilian hosts, particularly Anolis lizards. Full article
(This article belongs to the Special Issue Vectors and Vector-borne Diseases)
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11 pages, 1801 KiB  
Article
Detection of Leishmania and Trypanosoma DNA in Field-Caught Sand Flies from Endemic and Non-Endemic Areas of Leishmaniasis in Southern Thailand
by Pimpilad Srisuton, Atchara Phumee, Sakone Sunantaraporn, Rungfar Boonserm, Sriwatapron Sor-suwan, Narisa Brownell, Theerakamol Pengsakul and Padet Siriyasatien
Insects 2019, 10(8), 238; https://doi.org/10.3390/insects10080238 - 2 Aug 2019
Cited by 32 | Viewed by 4672
Abstract
Phlebotomine sand flies are tiny, hairy, blood-sucking nematoceran insects that feed on a wide range of hosts. They are known as a principal vector of parasites, responsible for human and animal leishmaniasis worldwide. In Thailand, human autochthonous leishmaniasis and trypanosomiasis have been reported. [...] Read more.
Phlebotomine sand flies are tiny, hairy, blood-sucking nematoceran insects that feed on a wide range of hosts. They are known as a principal vector of parasites, responsible for human and animal leishmaniasis worldwide. In Thailand, human autochthonous leishmaniasis and trypanosomiasis have been reported. However, information on the vectors for Leishmania and Trypanosoma in the country is still limited. Therefore, this study aims to detect Leishmania and Trypanosoma DNA in field-caught sand flies from endemic areas (Songkhla and Phatthalung Provinces) and non-endemic area (Chumphon Province) of leishmaniasis. A total of 439 sand flies (220 females and 219 males) were collected. Head and genitalia dissection of female sandflies were done for morphology identification, and the remaining parts of those sand flies were then used for the detection of Leishmania and Trypanosoma parasites. The DNA was extracted from individual female sand flies. Polymerase chain reaction (PCR) anneal, specific to the ITS1 and SSU rRNA gene regions, was used to detect Leishmania and Trypanosoma DNA, respectively. The positive PCR products were cloned and sequenced. The results showed that the female sand fly species in this study consisted of Sergentomyia khawi (35.9%); Se. anodontis (23.6%); Phlebotomus betisi (18.6%); Ph. kiangsuensis (9.5%); Ph. asperulus (6.4%); Se. barraudi (2.3%); 0.9% of each Se. indica, Ph. stantoni, and Ph. major major; and 0.5% of each Se. sylvatica and Ph. mascomai. The PCR and sequence analysis were able to detect Leishmania and Trypanosoma DNA in sand fly samples, which were identified as L. martiniquensis, 1/220 (0.45%) in Se. khawi, 3/220 (1.36%) of T. noyesi in Se. anodontis, and Ph. asperulus. Fourteen (6.36%) of the unidentified trypanosome species in Se. khawi, Se. indica, Se. anodontis, Ph. asperulus, and Ph. betisi were found in all of the areas of this study. Interestingly, we found a 1/220 (0.45%) co-infection sample of L. martiniquensis and Trypanosoma in Se. khawi from Songkhla Province. These data indicate that several species of sand flies might be potential vectors of Leishmania and Trypanosoma parasites in southern Thailand. However, more extensive study for potential vectors using a larger number of sand flies should be conducted to prove whether these sand flies can be natural vectors of leishmaniasis and trypanosomiasis in both humans and animals. In addition, our study could be useful for the future study of infection prevention, including effective vector control for leishmaniasis and trypanosomiasis in Thailand. Full article
(This article belongs to the Special Issue Vectors and Vector-borne Diseases)
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15 pages, 2353 KiB  
Article
Aedes aegypti Males as Vehicles for Insecticide Delivery
by Corey L. Brelsfoard, James W. Mains, Steve Mulligan, Anthony Cornel, Jodi Holeman, Susanne Kluh, Andrea Leal, Lawrence J. Hribar, Harold Morales, Tanya Posey and Stephen L. Dobson
Insects 2019, 10(8), 230; https://doi.org/10.3390/insects10080230 - 1 Aug 2019
Cited by 15 | Viewed by 4122
Abstract
Aedes aegypti continues to spread globally and remains a challenge to control, in part due to its ‘cryptic behavior’ in that it often deposits eggs (oviposits) in larval habitats that are difficult to find and treat using traditional methods. Auto-dissemination strategies target these [...] Read more.
Aedes aegypti continues to spread globally and remains a challenge to control, in part due to its ‘cryptic behavior’ in that it often deposits eggs (oviposits) in larval habitats that are difficult to find and treat using traditional methods. Auto-dissemination strategies target these cryptic breeding sites by employing mosquitoes to deliver lethal doses of insecticide. This report describes the initial field trials of an application known as Autodissemination Augmented by Males (ADAM), utilizing A. aegypti males dusted with pyriproxyfen (PPF). Findings presented here are drawn from both caged and field trial studies. Together, these trials examined for the ability of A. aegypti males to disseminate PPF and to impact field populations. PPF-dusted males were able to effectively deliver lethal doses of PPF to oviposition sites under the conditions tested. Results from field trials in Florida and California demonstrated reduced A. aegypti populations in treated areas, compared to areas where PPF-treated males were not released. These results indicate that the release of PPF-dusted A. aegypti males can impact A. aegypti populations as measured by both reduced larval survival and lower numbers of adult female A. aegypti. We propose the ADAM approach as an addition to existing mosquito control techniques targeting A. aegypti and other mosquitoes that utilize cryptic larval habitats. Full article
(This article belongs to the Special Issue Vectors and Vector-borne Diseases)
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10 pages, 2805 KiB  
Article
High Prevalence of a Newly Discovered Wutai Mosquito Phasivirus in Mosquitoes from Rio de Janeiro, Brazil
by Mário Sérgio Ribeiro, Tania Ayllón, Viviana Malirat, Daniel Cardoso Portela Câmara, Cristina Maria Giordano Dias, Guilherme Louzada, Davis Fernandes-Ferreira, Roberto de Andrade Medronho and Renata Campos Acevedo
Insects 2019, 10(5), 135; https://doi.org/10.3390/insects10050135 - 7 May 2019
Cited by 4 | Viewed by 3078
Abstract
Many RNA viruses have recently emerged, threatening humans and causing harm to animals and plants. Bunyaviruses represent one of the largest groups of RNA viruses and are able to infect a wide range of hosts (invertebrates, vertebrates, and plants). Recently, new insect-specific viruses [...] Read more.
Many RNA viruses have recently emerged, threatening humans and causing harm to animals and plants. Bunyaviruses represent one of the largest groups of RNA viruses and are able to infect a wide range of hosts (invertebrates, vertebrates, and plants). Recently, new insect-specific viruses have been isolated from mosquitoes and phlebotomine sandflies worldwide. Little is known regarding the impact of these viruses on the vector life cycles and the stages of oviposition, breeding, blood feeding, and the mosquito’s lifespan. This study describes, for the first time in South America, the detection and characterization of a recently discovered bunyavirus corresponding to the Wutai mosquito phasivirus, confirming its high prevalence in the Culex spp. and Aedes spp. mosquitoes collected in the urban environment of Rio de Janeiro city, Brazil. The knowledge of the mosquito’s insect-specific virus infection can improve virus evolution studies and may contribute to the understanding of intrinsic factors that influence vector competence to transmit pathogenic viruses. Full article
(This article belongs to the Special Issue Vectors and Vector-borne Diseases)
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Review

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25 pages, 1132 KiB  
Review
Biological Adaptations Associated with Dehydration in Mosquitoes
by Christopher J. Holmes and Joshua B. Benoit
Insects 2019, 10(11), 375; https://doi.org/10.3390/insects10110375 - 28 Oct 2019
Cited by 18 | Viewed by 7121
Abstract
Diseases that are transmitted by mosquitoes are a tremendous health and socioeconomic burden with hundreds of millions of people being impacted by mosquito-borne illnesses annually. Many factors have been implicated and extensively studied in disease transmission dynamics, but knowledge regarding how dehydration impacts [...] Read more.
Diseases that are transmitted by mosquitoes are a tremendous health and socioeconomic burden with hundreds of millions of people being impacted by mosquito-borne illnesses annually. Many factors have been implicated and extensively studied in disease transmission dynamics, but knowledge regarding how dehydration impacts mosquito physiology, behavior, and resulting mosquito-borne disease transmission remain underdeveloped. The lapse in understanding on how mosquitoes respond to dehydration stress likely obscures our ability to effectively study mosquito physiology, behavior, and vectorial capabilities. The goal of this review is to develop a profile of factors underlying mosquito biology that are altered by dehydration and the implications that are related to disease transmission. Full article
(This article belongs to the Special Issue Vectors and Vector-borne Diseases)
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Other

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7 pages, 603 KiB  
Brief Report
RNAi by Soaking Aedes aegypti Pupae in dsRNA
by Fiza Arshad, Arvind Sharma, Charleen Lu and Monika Gulia-Nuss
Insects 2021, 12(7), 634; https://doi.org/10.3390/insects12070634 - 13 Jul 2021
Cited by 5 | Viewed by 3520
Abstract
RNA-interference (RNAi) is a standard technique for functional genomics in adult mosquitoes. However, RNAi in immature, aquatic mosquito stages has been challenging. Several studies have shown successful larval RNAi, usually in combination with a carrier molecule. Except for one study in malaria mosquito, [...] Read more.
RNA-interference (RNAi) is a standard technique for functional genomics in adult mosquitoes. However, RNAi in immature, aquatic mosquito stages has been challenging. Several studies have shown successful larval RNAi, usually in combination with a carrier molecule. Except for one study in malaria mosquito, Anopheles gambiae, none of the previous studies has explored RNAi in mosquito pupae. Even in the study that used RNAi in pupae, double stranded RNA (dsRNA) was introduced by microinjection. Here, we describe a successful method by soaking pupae in water containing dsRNA without any carrier or osmotic challenge. The knockdown persisted into adulthood. We expect that this simple procedure will be useful in the functional analysis of genes that highly express in pupae or newly emerged adults. Full article
(This article belongs to the Special Issue Vectors and Vector-borne Diseases)
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