scholarly journals Cas9-Mediated Gene-Editing in the Malaria Mosquito Anopheles stephensi by ReMOT Control

2020 ◽  
Vol 10 (4) ◽  
pp. 1353-1360 ◽  
Author(s):  
Vanessa M. Macias ◽  
Sage McKeand ◽  
Duverney Chaverra-Rodriguez ◽  
Grant L. Hughes ◽  
Aniko Fazekas ◽  
...  
Keyword(s):  
Gene Editing ◽  
Molecular Mechanisms ◽  
Anopheles Stephensi ◽  
Mosquito Control ◽  
Control Strategies ◽  
Mosquito Species ◽  
Adult Mosquito ◽  
Malaria Mosquito ◽  
Powerful Method ◽  
Injection Methods

Innovative tools are essential for advancing malaria control and depend on an understanding of molecular mechanisms governing transmission of malaria parasites by Anopheles mosquitoes. CRISPR/Cas9-based gene disruption is a powerful method to uncover underlying biology of vector-pathogen interactions and can itself form the basis of mosquito control strategies. However, embryo injection methods used to genetically manipulate mosquitoes (especially Anopheles) are difficult and inefficient, particularly for non-specialist laboratories. Here, we adapted the ReMOT Control (Receptor-mediated Ovary Transduction of Cargo) technique to deliver Cas9 ribonucleoprotein complex to adult mosquito ovaries, generating targeted and heritable mutations in the malaria vector Anopheles stephensi without injecting embryos. In Anopheles, ReMOT Control gene editing was as efficient as standard embryo injections. The application of ReMOT Control to Anopheles opens the power of CRISPR/Cas9 methods to malaria laboratories that lack the equipment or expertise to perform embryo injections and establishes the flexibility of ReMOT Control for diverse mosquito species.

scholarly journals Cas9-mediated gene-editing in the malaria mosquito Anopheles stephensi by ReMOT Control

2019 ◽  
Author(s):  
Vanessa M. Macias ◽  
Sage McKeand ◽  
Duverney Chaverra-Rodriguez ◽  
Grant L. Hughes ◽  
Aniko Fazekas ◽  
...  
Keyword(s):  
Malaria Vector ◽  
Gene Editing ◽  
Reverse Genetics ◽  
Molecular Mechanisms ◽  
Anopheles Stephensi ◽  
Control Strategies ◽  
Adult Mosquito ◽  
Pathogen Control ◽  
Innovative Tool ◽  
Injection Methods

AbstractInnovative tool development is essential for continued advancement in malaria control and depends on a deeper understanding of the molecular mechanisms that govern transmission of malaria parasites by Anopheles mosquitoes. Targeted disruption of genes in mosquito vectors is a powerful method to uncover the underlying biology of vector-pathogen interactions, and genome manipulation technologies can themselves form the basis of mosquito and pathogen control strategies. However, the embryo injection methods used to genetically manipulate mosquitoes, and in particular Anopheles species, are difficult and inefficient, particularly for non-specialist laboratories. We have adapted a strategy called ReMOT Control (Receptor-mediated Ovary Transduction of Cargo) to deliver the Cas9 ribonucleoprotein complex to adult mosquito ovaries and generate targeted and heritable mutations in the malaria vector Anopheles stephensi. We found that gene editing by ReMOT Control in Anopheles mosquitoes was comparable to the technique in Ae. aegypti and as efficient in editing as standard embryo injections. The adaptation of this technology to Anopheles mosquitoes opens up the power of reverse genetics to malaria vector labs that do not have the equipment or technical expertise to perform embryo injections and establishes the flexibility of ReMOT Control for gene-editing in non-Aedes species.

scholarly journals Inhibition of Asaia in Adult Mosquitoes Causes Male-Specific Mortality and Diverse Transcriptome Changes

Pathogens ◽  
2020 ◽  
Vol 9 (5) ◽  
pp. 380 ◽  
Author(s):  
Maria Vittoria Mancini ◽  
Claudia Damiani ◽  
Sarah M. Short ◽  
Alessia Cappelli ◽  
Ulisse Ulissi ◽  
...  
Keyword(s):  
Mosquito Control ◽  
Control Strategies ◽  
Mosquito Species ◽  
Acetic Acid Bacteria ◽  
Adult Mosquito ◽  
Great Expectations ◽  
Innovative Tool ◽  
Male Specific

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.

scholarly journals Mosquito Control Priorities in Florida—Survey Results from Florida Mosquito Control Districts

Pathogens ◽  
2021 ◽  
Vol 10 (8) ◽  
pp. 947
Author(s):  
Rishi Kondapaneni ◽  
Ashley N. Malcolm ◽  
Brian M. Vazquez ◽  
Eric Zeng ◽  
Tse-Yu Chen ◽  
...  
Keyword(s):  
State Government ◽  
Mosquito Control ◽  
Control Strategies ◽  
Mosquito Species ◽  
Research Priorities ◽  
Current Control ◽  
Control Programs ◽  
Common Control ◽  
Surveillance Programs ◽  
Research Questions

Florida lies within a subtropical region where the climate allows diverse mosquito species including invasive species to thrive year-round. As of 2021, there are currently 66 state-approved Florida Mosquito Control Districts, which are major stakeholders for Florida public universities engaged in mosquito research. Florida is one of the few states with extensive organized mosquito control programs. The Florida State Government and Florida Mosquito Control Districts have long histories of collaboration with research institutions. During fall 2020, we carried out a survey to collect baseline data on the current control priorities from Florida Mosquito Control Districts relating to (1) priority control species, (2) common adult and larval control methods, and (3) major research questions to address that will improve their control and surveillance programs. The survey data showed that a total of 17 distinct mosquito species were considered to be priority control targets, with many of these species being understudied. The most common control approaches included truck-mounted ultra-low-volume adulticiding and biopesticide-based larviciding. The districts held interest in diverse research questions, with many prioritizing studies on basic science questions to help develop evidence-based control strategies. Our data highlight the fact that mosquito control approaches and priorities differ greatly between districts and provide an important point of comparison for other regions investing in mosquito control, particularly those with similar ecological settings, and great diversity of potential mosquito vectors, such as in Florida. Our findings highlight a need for greater alignment of research priorities between mosquito control and mosquito research. In particular, we note a need to prioritize filling knowledge gaps relating to understudied mosquito species that have been implicated in arbovirus transmission.

scholarly journals Mosquitoes of Etiological Concern in Kenya and Possible Control Strategies

Insects ◽  
2019 ◽  
Vol 10 (6) ◽  
pp. 173 ◽  
Author(s):  
Samuel Karungu ◽  
Evans Atoni ◽  
Joseph Ogalo ◽  
Caroline Mwaliko ◽  
Bernard Agwanda ◽  
...  
Keyword(s):  
Environmental Changes ◽  
Mosquito Control ◽  
Control Strategies ◽  
Mosquito Species ◽  
Disease Outbreaks ◽  
Health Concern ◽  
Public Health Concern ◽  
Vector Management ◽  
Invasive Mosquito ◽  
Parasitic Pathogens

Kenya is among the most affected tropical countries with pathogen transmitting Culicidae vectors. For decades, insect vectors have contributed to the emergence and distribution of viral and parasitic pathogens. Outbreaks and diseases have a great impact on a country’s economy, as resources that would otherwise be used for developmental projects are redirected to curb hospitalization cases and manage outbreaks. Infected invasive mosquito species have been shown to increasingly cross both local and global boarders due to the presence of increased environmental changes, trade, and tourism. In Kenya, there have been several mosquito-borne disease outbreaks such as the recent outbreaks along the coast of Kenya, involving chikungunya and dengue. This certainly calls for the implementation of strategies aimed at strengthening integrated vector management programs. In this review, we look at mosquitoes of public health concern in Kenya, while highlighting the pathogens they have been linked with over the years and across various regions. In addition, the major strategies that have previously been used in mosquito control and what more could be done to reduce or combat the menace caused by these hematophagous vectors are presented.

scholarly journals Molecular Characterization of Culturable Aerobic Bacteria in the Midgut of Field-Caught Culex tritaeniorhynchus, Culex gelidus, and Mansonia annulifera Mosquitoes in the Gampaha District of Sri Lanka

2020 ◽  
Vol 2020 ◽  
pp. 1-13
Author(s):  
Nayana Gunathilaka ◽  
Koshila Ranasinghe ◽  
Deepika Amarasinghe ◽  
Wasana Rodrigo ◽  
Harendra Mallawarachchi ◽  
...  
Keyword(s):  
Sri Lanka ◽  
Vector Control ◽  
Control Strategies ◽  
Mosquito Species ◽  
Diversity Indices ◽  
Plate Count ◽  
Adult Mosquito ◽  
Relative Distribution ◽  
Bacterial Isolates ◽  
Culex Tritaeniorhynchus

Background. Larval and adult mosquito stages harbor different extracellular microbes exhibiting various functions in their digestive tract including host-parasite interactions. Midgut symbiotic bacteria can be genetically exploited to express molecules within the vectors, altering vector competency and potential for disease transmission. Therefore, identification of mosquito gut inhabiting microbiota is of ample importance before developing novel vector control strategies that involve modification of vectors. Method. Adult mosquitoes of Culex tritaeniorhynchus, Culex gelidus, and Mansonia annulifera were collected from selected Medical Officer of Health (MOH) areas in the Gampaha district of Sri Lanka. Midgut lysates of the field-caught non-blood-fed female mosquitoes were cultured in Plate Count Agar medium, and Prokaryotic 16S ribosomal RNA partial genes of the isolated bacteria colonies were amplified followed by DNA sequencing. Diversity indices were used to assess the diversity and richness of the bacterial isolates in three mosquito species. The distribution pattern of bacterial isolates between different mosquito species was assessed by Distance-Based Redundancy Analysis (dbRDA). Results. A total of 20 bacterial species (Staphylococcus pasteuri, Bacillus megaterium, Staphylococcus cohnii, Pantoea dispersa, Staphylococcus chromogenes, Bacillus aquimaris, Staphylococcus arlettae, Staphylococcus sciuri, Staphylococcus warneri, Moraxella osloensis, Enterobacter sp., Klebsiella michiganensis, Staphylococcus hominis, Staphylococcus saprophyticus, Streptomyces sp., Bacillus niacin, Cedecea neteri, Micrococcus luteus, Lysinibacillus sphaericus, and Bacillus licheniformis) were identified. All of these species belonged to three phyla, Proteobacteria, Firmicutes, and Actinobacteria, out of which phylum Firmicutes (71.1%) was the most prominent. The least number of species was recorded from Actinobacteria. The relative distribution of midgut microbes in different mosquito species differed significantly among mosquito species (Chi-square, χ2=486.091; df=36; P≤0.001). Midgut microbiota of Cx. tritaeniorhynchus and Cx. gelidus indicated a similarity of 21.51%, while Ma. annulifera shared a similarity of 6.92% with the cluster of above two species. The gut microbiota of Cx. tritaeniorhynchus was also significantly more diverse and more evenly distributed compared to Ma. annulifera. Simpson’s diversity, Margalef’s diversity, and Menhinick’s diversity indices were higher in Cx. gelidus. Of the recorded species, P. dispersa and strains of nonpathogenic species in Bacillaceae family (B. megaterium, B. niacini, B. licheniformis, and L. sphaericus) can be recommended as potential candidates for paratransgenesis. Conclusion. The relative distribution of midgut microbes in different mosquito species differed significantly among the three studied adult mosquito species. The present data strongly encourage further investigations to explore the potential usage of these microbes through paratransgenic approach for novel eco-friendly vector control strategies.

scholarly journals An overview on nano-materials as mosquito controlling agents against Anopheles stephensi

2020 ◽  
Vol 4 (3) ◽  
Keyword(s):  
Urban Areas ◽  
Anopheles Stephensi ◽  
Mosquito Control ◽  
Control Strategies ◽  
Cost Effective ◽  
Control Agent ◽  
Mosquito Vector ◽  
Nano Materials ◽  
Vector Borne Diseases ◽  
Plant Parts

Mosquitoes are very obnoxious insects and vectors of many diseases like malaria, filariasis, dengue, yellow fever, encephalitis etc. Among these, malaria is transmitted by Anopheles stephensi mainly in urban areas across the temperate country. So, there is an imperative requirement to imply some mechanisms to reduce the mosquito vector to facilitate the reduction of vector borne diseases throughout the globe. But, mosquito control strategies are now in front of a threat because mosquito populations get resistance against the available synthetic insecticides. Besides the chemical insecticides, several other ways to control the vector population proved unsuccessful in most of the cases. Now, various researchers are trying to develop an alternate effective approach to eradicate malaria from the earth by controlling its vector by using a product which is ecofreindly, cost effective, biodegradable, non-toxic and selective in nature. So, by the amalgamation of the two fields namely the nanotechnology and the vector managements, a new field is now opened up for decades, “the green synthesized nanoparticles as mosquito control agents”. The plant-based, fabricated nanoparticles are beneficial over the synthetic one since it is economical, eco-friendly and non-toxic to nature. Silver, gold, zinc, cobalt, copper etc. nanoparticles are green fabricated with different plant parts, fungi as well as bacteria. These nano-materials are of less than 100 nm in diameter and are currently used in various fields of industry. So, adapting the nano-biotechnology to control the mosquito vector is the need of the hour. In this present review we focused on potential role of nano-materials as mosquito control agent against An. stephensi at different life stages of it.

scholarly journals Genome Features of Asaia sp. W12 Isolated from the Mosquito Anopheles stephensi Reveal Symbiotic Traits

Genes ◽  
2021 ◽  
Vol 12 (5) ◽  
pp. 752
Author(s):  
Shicheng Chen ◽  
Ting Yu ◽  
Nicolas Terrapon ◽  
Bernard Henrissat ◽  
Edward D. Walker
Keyword(s):  
Molecular Mechanisms ◽  
Anopheles Stephensi ◽  
Environmental Changes ◽  
Plant Biomass ◽  
Close Association ◽  
Mosquito Species ◽  
Regulatory Protein ◽  
Terminal Oxidases ◽  
Multiple Organs ◽  
Insect Gut

Asaia bacteria commonly comprise part of the microbiome of many mosquito species in the genera Anopheles and Aedes, including important vectors of infectious agents. Their close association with multiple organs and tissues of their mosquito hosts enhances the potential for paratransgenesis for the delivery of antimalaria or antivirus effectors. The molecular mechanisms involved in the interactions between Asaia and mosquito hosts, as well as Asaia and other bacterial members of the mosquito microbiome, remain underexplored. Here, we determined the genome sequence of Asaia strain W12 isolated from Anopheles stephensi mosquitoes, compared it to other Asaia species associated with plants or insects, and investigated the properties of the bacteria relevant to their symbiosis with mosquitoes. The assembled genome of strain W12 had a size of 3.94 MB, the largest among Asaia spp. studied so far. At least 3585 coding sequences were predicted. Insect-associated Asaia carried more glycoside hydrolase (GH)-encoding genes than those isolated from plants, showing their high plant biomass-degrading capacity in the insect gut. W12 had the most predicted regulatory protein components comparatively among the selected Asaia, indicating its capacity to adapt to frequent environmental changes in the mosquito gut. Two complete operons encoding cytochrome bo3-type ubiquinol terminal oxidases (cyoABCD-1 and cyoABCD-2) were found in most Asaia genomes, possibly offering alternative terminal oxidases and allowing the flexible transition of respiratory pathways. Genes involved in the production of 2,3-butandiol and inositol have been found in Asaia sp. W12, possibly contributing to biofilm formation and stress tolerance.

scholarly journals Do tigers hunt during the day? Diel activity of the Asian tiger mosquito, Aedes albopictus (Diptera: Culicidae), in urban and suburban habitats of North America

2021 ◽  
Vol 15 (8) ◽  
pp. e0009438
Author(s):  
Isik Unlu ◽  
Ary Faraji ◽  
Nicholas Indelicato ◽  
James R. McNelly
Keyword(s):  
Public Health ◽  
Aedes Albopictus ◽  
Mosquito Control ◽  
Mosquito Species ◽  
Diel Activity ◽  
Adult Mosquito ◽  
Subtropical Climate ◽  
Public Health Importance ◽  
Late Season ◽  
Solar Noon

Background Aedes (Stegomyia) albopictus (Skuse) impacts human outdoor activity because of its aggressive biting behavior, and as a major vector of mosquito-borne diseases, it is also of public health importance. Although most mosquito species exhibit crepuscular activity by primarily host seeking at dawn and dusk, Ae. albopictus has been traditionally characterized as a diurnal or day-biting mosquito. With the global expansion and increased involvement of Ae. albopictus in mosquito-borne diseases, it is imperative to elucidate the diel activity of this species, particularly in newly invaded areas. Methodology and principal findings Human sweep netting and carbon dioxide-baited rotator traps were used to evaluate the diel activity of Ae. albopictus in two study sites. Both trapping methods were used in New Jersey’s Mercer County, USA (temperate/urban), while only human sweep netting was used in Florida’s Volusia County, USA (subtropical/suburban). Human sweep netting was performed to determine adult mosquito activity at Sunrise, Solar Noon, Sunset, and Lunar Midnight. Because New Jersey is in a temperate area, diel activity was investigated during the early season (3–19 July), peak season (25 July-19 September), and late season (22 September- 22 October). Aedes albopictus showed the highest activity during peak and late seasons at Solar Noon (P < 0.05). At Sunrise and Sunset during the peak season, Ae. albopictus activity was similar. Lunar Midnight activity was significantly lower than Sunrise and Solar Noon (P < 0.05) but was similar to that of Sunset. In the late season, the highest activity was observed during Solar Noon while the least activity was observed during Sunrise and Lunar Midnight (P<0.05). Bottle rotator traps used in conjunction with the human sweep net technique exhibited similar results. Seasonal activity was not differentiated in Florida due to the consistent subtropical climate. The highest adult activity was observed at Sunrise using human sweep netting, but it was not significantly different from Solar Noon and Sunset. The lowest adult activity was observed at Lunar Midnight; however, it was not significantly different from Solar Noon and Sunset. These results provide evidence that the diel activity of Ae. albopictus, contrary to the common perception of its diurnal activity, is much more varied. Conclusion/Significance Involvement of Ae. albopictus in the transmission of debilitating mosquito-borne pathogens such as chikungunya, dengue, and Zika virus, coupled with its affinity to thrive in human peridomestic environments, substantiates that our findings have global implications in areas where Ae. albopictus populations established. It also highlights the importance of behavioral studies of vector species which will not only help mosquito control professionals plan the timing of their control efforts but also provides empirical evidence against conventional wisdoms that may unjustly persist within public health stewards.

scholarly journals Trend of mosquito (Diptera: Culicidae) monthly distribution in Sanandaj County of Iran

2021 ◽  
Vol 22 (11) ◽  
Author(s):  
Bahadin Salavati ◽  
AMIR HOSSEIN ZAHIRNIA ◽  
Hassan Nasirian ◽  
SHAHYAD AZARI-HAMIDIAN
Keyword(s):  
Urban Areas ◽  
Mosquito Control ◽  
Mosquito Species ◽  
Control Measures ◽  
Adult Mosquito ◽  
Rural And Urban Areas ◽  
Rural And Urban ◽  
Significant Difference ◽  
Monthly Distribution ◽  
Mosquito Breeding

Abstract. Salavati B, Zahirnia AH, Nasirian H, Hamidian SA. 2021. Trend of mosquito (Diptera: Culicidae) monthly distribution in Sanandaj County of Iran. Biodiversitas 22: 4705-4715. Mosquitoes are the most significant arthropods of medical importance due to their burden of transmitting pathogens and parasites. The present study was designed to investigate the distribution of mosquitos (Diptera: Culicidae) in Sanandaj County in 2018. Sanandaj County was divided into four areas in rural and urban areas, including north, south, east, and west. Four rural and urban areas were randomly selected from four geographical directions. A sampling of larval and adult mosquitoes was performed using standard methods. The dipping technique was used to collect the mosquito larvae. A sampling of adult mosquitoes was performed during feeding on humans and animal baits and resting in the natural indoors, outdoors, and artificial areas. Light traps were also used to collect adult mosquitoes. The larval and adult specimens were identified using the morphology-based key. In total, nine mosquito species, including Anopheles maculipennis s.l., Anopheles superpictus s.l., Culiseta longiareolata, Culiseta subochrea, Culex hortensis, Culex mimeticus, Culex modestus, Culex pipiens, and Culex theileri were inhabited in Sanandaj County. The larval and adult mosquito species trend remains constant for at least 6 months of a year, from May to October. One-way ANOVA didn't reveal a significant difference between mosquito breeding places' habitats and water types, mosquito localities and life stages, monthly mosquito distribution, and adult mosquito resting places. The frequency trends of larval and adult mosquito species remain constant at least for 6 months of a year. It needs to be ready for timely mosquito control measures.

scholarly journals Genome Features of Asaia sp. W12 Isolated from the Mosquito Anopheles stephensi Reveal Symbiotic Traits

2021 ◽  
Author(s):  
Shicheng Chen ◽  
Ting Yu ◽  
Nicolas Terrapon ◽  
Bernard Henrissat ◽  
Edward Walker
Keyword(s):  
Molecular Mechanisms ◽  
Anopheles Stephensi ◽  
Environmental Changes ◽  
Close Association ◽  
Mosquito Species ◽  
Regulatory Protein ◽  
Terminal Oxidases ◽  
Cytochrome Bo3 ◽  
Multiple Organs ◽  
Protein Components

Asaia bacteria commonly comprise part of the microbiome of many mosquito species in the genera Anopheles and Aedes, including important vectors of infectious agents. Their close association with multiple organs and tissues of their mosquito hosts enhances the potential for paratransgenesis for delivery of anti-malaria or anti-virus effectors. The molecular mechanisms involved in the interactions between Asaia and mosquito hosts, as well as Asaia and other bacterial members of the mosquito microbiome, remained unexplored. Here, we determined the genome sequence of the strain W12 isolated from Anopheles stephensi mosquitoes, compared them to other Asaia species associated with plants or insects, and investigated some properties of the bacteria relevant to their symbiosis with host mosquitoes. The assembled genome of strain W12 has a size of 3.94 MB, which is the largest among Asaia spp studied so far. At least 3,585 coding sequences were predicted. The insect-associated Asaia including strain W12 carried more glycoside hydrolase (GH) encoding genes (31 per genome) than those isolated from plants (22 per genome). W12 had the most predicted regulatory protein components (213) among the selected Asaia (ranging from 131 to 211), indicating its great capability to adapt to frequent environmental changes in the mosquito gut. Two complete operons encoding cytochrome bo3-type ubiquinol terminal oxidases (cyoABCD-1 and cyoABCD-2) were found in most of Asaia genomes, which possibly offer alternative terminal oxidases and allow the flexible transition of respiratory pathways. Genes involved in the production of acetoin and 2,3-butandiol have been identified in Asaia sp. W12.

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