Keywords: Arbovirus
Wolbachia as a transformative tool for mosquito-borne disease control: a comprehensive review of mechanisms, efficacy, and future directions
35325Minaei, M. E., Yousefi Nojookambari, N., Ghodraty, M., & Yazdansetad, S., Pathogens and Global Health, 2025-12-23 12:15:44.
Mosquito-borne infectious diseases such as dengue, Zika, chikungunya, and malaria continue to pose significant public health challenges worldwide. Traditional control methods, including the use of insecticides and environmental management, have shown limited effectiveness due to insecticide resistance and logistical challenges. Wolbachia-based strategies, which leverage the bacterium’s unique ability to manipulate mosquito reproduction and block pathogens, present a promising, albeit not universally applicable, method for reducing the transmission of mosquito-borne diseases. Field trials have demonstrated significant but variable efficacy. This systematic review synthesizes the latest findings on Wolbachia-based interventions, focusing on their mechanisms, efficacy, and potential for large-scale implementation. We conducted a comprehensive literature search across multiple databases, including PubMed, Scopus, and Web of Science, to identify relevant studies published between 1990 and 2025. Our review highlights the promising results of Wolbachia-based strategies in reducing mosquito populations and disease transmission, while also addressing the challenges and limitations of this approach. We conclude that Wolbachia-based strategies hold great potential for revolutionizing mosquito-borne disease control, but further research and careful consideration of ecological and ethical implications are necessary for their successful implementation.
Rethinking the future of mosquito control
35341Kabirul Bashar, Dr Zonaed Siddiki. Dr Zonaed Siddiki, et al., Dhaka Tribune, 2025-12-17 11:37:26.
Bangladesh is once again in the grip of a severe dengue and chikungunya crisis in 2025. Hospitals are overwhelmed, fever wards are full, and doctors are working tirelessly. Families live in fear of mosquito bites, while daily news reports chronicle rising infections and deaths. As of December 16, 2025, the Directorate General of Health Services (DGHS) has reported more than 100,000 confirmed dengue cases and 409 deaths. However, this figure represents only the tip of the iceberg, as it reflects data solely from hospitalized patients, specifically from 77 hospitals in Dhaka and reports from 64 civil surgeon offices. A substantial number of dengue patients are receiving treatment at home or in various small and large hospitals and clinics that are not included in the official count. This underscores the overwhelming pressure on the country’s health system. At the same time, the Institute of Epidemiology, Disease Control and Research (IEDCR) has recorded a resurgence of chikungunya, reporting 337 suspected cases, 153 of which were laboratory-confirmed in Dhaka between January and May 2025. Researchers estimate that the true number of chikungunya infections this year may be close to one hundred thousand. As chikungunya testing is available only in a limited number of major hospitals in Dhaka, many cases remain undiagnosed and unreported.
Bacteria-mediated dsRNA delivery for mosquito-borne virus control
35219Mine Altinli, Sean P. Leonard, Alain Kohl, et al., Trends in Parasitology, 41:880-893. 2025-10-05 14:37:07.
Mosquito-borne viruses represent an increasing global public health threat, exacerbated by urbanisation and climate change, thus making effective mosquito control essential. RNA interference (RNAi), a sequence-specific gene regulation mechanism, can be a flexible vector control tool. RNAi effectors, such as double-stranded RNA (dsRNA), can target mosquito genes or the viruses they carry, disrupting development or suppressing infection. However, current RNAi delivery methods are ineffective. Engineered bacterial symbionts offer a promising alternative for delivery, as they can produce dsRNA directly within mosquitoes. However, bacterial RNAi delivery in mosquitoes remains underexplored. We review emerging genetic tools, insights from RNAi and bacteria–mosquito interactions to outline priorities for realising bacterial RNAi as an efficient and sustainable vector control strategy.
Implications of successive blood feeding on Wolbachia-mediated dengue virus inhibition in Aedes aegypti mosquitoes
34989Johnson, R.M., Breban, M.I., Nolan, B.L. et al., Nat Commun, 16. 2025-07-30 10:55:57.
Wolbachia is a promising strategy to inhibit dengue virus (DENV) transmission by Ae. aegypti mosquitoes. Laboratory studies assessing DENV inhibition by Wolbachia typically have not considered natural frequent mosquito blood feeding behavior. Here, we determine the impact of successive feeding on DENV-2 transmission by Ae. aegypti in the presence or absence of Wolbachia (wAlbB and wMelM strains). We show that successive feeding shortens the extrinsic incubation period (EIP) in wildtype (WT; without Wolbachia) and wAlbB mosquitoes through enhanced dissemination. Feeding empirical data into models showed that successive feeding increases the probability of WT and wAlbB mosquitoes surviving beyond the EIP. Importantly, the more epidemiologically relevant comparison of the odds of wAlbB mosquitoes surviving beyond the EIP relative to WT, reveals a larger impact of successive feeding on WT than wAlbB. This indicates a strong inhibitory effect of Wolbachia even in the context of natural frequent mosquito blood feeding behavior.
Exploiting Wolbachia as a Tool for Mosquito-Borne Disease Control: Pursuing Efficacy, Safety, and Sustainability
34677Moretti, R., Lim, J. T., Ferreira, A. G. A., et al, Pathogens, 14:285. 2025-03-28 09:41:16.
Despite the application of control measures, mosquito-borne diseases continue to pose a serious threat to human health. In this context, exploiting Wolbachia, a common symbiotic bacterium in insects, may offer effective solutions to suppress vectors or reduce their competence in transmitting several arboviruses. Many Wolbachia strains can induce conditional egg sterility, known as cytoplasmic incompatibility (CI), when infected males mate with females that do not harbor the same Wolbachia infection. Infected males can be mass-reared and then released to compete with wild males, reducing the likelihood of wild females encountering a fertile mate. Furthermore, certain Wolbachia strains can reduce the competence of mosquitoes to transmit several RNA viruses. Through CI, Wolbachia-infected individuals can spread within the population, leading to an increased frequency of mosquitoes with a reduced ability to transmit pathogens. Using artificial methods, Wolbachia can be horizontally transferred between species, allowing the establishment of various laboratory lines of mosquito vector species that, without any additional treatment, can produce sterilizing males or females with reduced vector competence, which can be used subsequently to replace wild populations. This manuscript reviews the current knowledge in this field, describing the different approaches and evaluating their efficacy, safety, and sustainability. Successes, challenges, and future perspectives are discussed in the context of the current spread of several arboviral diseases, the rise of insecticide resistance in mosquito populations, and the impact of climate change. In this context, we explore the necessity of coordinating efforts among all stakeholders to maximize disease control. We discuss how the involvement of diverse expertise—ranging from new biotechnologies to mechanistic modeling of eco-epidemiological interactions between hosts, vectors, Wolbachia, and pathogens—becomes increasingly crucial. This coordination is especially important in light of the added complexity introduced by Wolbachia and the ongoing challenges posed by global change.
Exploiting venom toxins in paratransgenesis to prevent mosquito-borne disease
34533French, S., Da Silva, R., Storm, J. et al., Parasites & Vectors, 18. 2025-02-21 11:33:53.
Mosquitoes are responsible for the transmission of numerous pathogens, including Plasmodium parasites, arboviruses and filarial worms. They pose a significant risk to public health with over 200 million cases of malaria per annum and approximately 4 billion people at risk of arthropod-borne viruses (arboviruses). Mosquito populations are geographically expanding into temperate regions and their distribution is predicted to continue increasing. Mosquito symbionts, including fungi, bacteria and viruses, have desirable traits for mosquito disease control including spreading horizontally and vertically through mosquito populations and potentially colonising multiple important vector species. Paratransgenesis, genetic modification of mosquito symbionts with effectors to target the pathogen rather than the vector, is a promising strategy to prevent the spread of mosquito-borne diseases. A variety of effectors can be expressed but venom toxins are excellent effector candidates because they are target specific, potent and stable. However, the only toxins to be explored in mosquito paratransgenesis to date are scorpine and mutated phospholipase A2. To enhance the scope, effectiveness and durability of paratransgenesis, an expanded arsenal of effectors is required. This review discusses other potential toxin effectors for future paratransgenesis studies based on prior in vitro and in vivo antiparasitic and antiviral studies and highlights the need for further research and investment in this area. In terms of mosquito-borne diseases, paratransgenesis strategies have been developed to target Plasmodium. We postulate the potential to apply this principle to target arboviruses using antiviral toxin effectors.
Man Vs. Mosquito
33147Aman Vora, Brown Political Review, 2024-10-31 08:49:52.
It is a middle school math teacher’s favorite trivia question: What is the world’s deadliest animal? After images of a hunting tiger or towering gorilla flash through our imagination, we remember that it is the humble mosquito, whose terrible impact on human lives and healthcare systems is only projected to grow. As carbon emissions continue to rise with no plateau in sight, one oft-forgotten implication of increasing global temperatures is the devastating impact they will wreak on public health, with historically ignored diseases now able to thrive in a new, warmer climate. As global temperatures barrel toward the preferred range for mosquitoes, the number of individuals at risk for contracting malaria and dengue fever may increase by four to seven billion by 2070 relative to 1999. This threat is already a reality. Take dengue, for example: From 1980 to 1989, there were 1.5 million reported cases globally. Compare that to 2019 alone, when 5.2 million cases were reported. World Health Organization (WHO) officials described this astronomical rise in dengue as a “canary in the coalmine of the climate crisis.” No longer will mosquito-borne diseases primarily threaten equatorial regions—northern cities globally are all at risk due to the rise of Aegypti and Anopheles mosquitoes. Science: 0. Mosquitoes: 1. From bed nets to insecticides, progress is being made to combat this terrifying rise. But the current generation of anti-mosquito tools is not aggressive enough to mitigate this deadly problem: Bed nets do little to stop Aegypti, which primarily feed on blood during the day, and toxic insecticides have done little but harm the environment and drive mosquito resistance. In order to save lives from this man-made and mosquito-driven catastrophe, humanity must embrace its most promising scientific technologies: genetic engineering and Wolbachia bacteria, conscious that we are fighting against both Mother Nature and human nature itself.
Wolbachia-based emerging strategies for control of vector-transmitted disease
32708Diego Montenegro, Gerardo Cortés-Cortés, María Guadalupe Balbuena-Alonso, et al., Acta Tropica, 260. 2024-10-22 17:28:54.
Dengue fever is a mosquito-transmitted disease of great public health importance. Dengue lacks adequate vaccine protection and insecticide-based methods of mosquito control are proving increasingly ineffective. Here we review the emerging use of mosquitoes transinfected with the obligate intracellular bacterium Wolbachia pipientis for vector control. Wolbachia often induces cytoplasmic incompatibility in its mosquito hosts, resulting in infertile progeny between an infected male and an uninfected female. Wolbachia infection also suppresses the replication of pathogens in the mosquito, a process known as “pathogen blocking”. Two strategies have emerged. The first one releases Wolbachia carriers (both male and female) to replace the wild mosquito population, a process driven by cytoplasmic incompatibility and that becomes irreversible once a threshold is reached. This suppresses disease transmission mainly by pathogen blocking and frequently requires a single intervention. The second strategy floods the field population with an exclusively male population of Wolbachia-carrying mosquitoes to generate infertile hybrid progeny. In this case, transmission suppression depends largely on decreasing the population density of mosquitoes driven by infertility and requires continued mosquito release. The efficacy of both Wolbachia-based approaches has been conclusively demonstrated by randomized and non-randomized studies of deployments across the world. However, results conducted in one setting cannot be directly or easily extrapolated to other settings because dengue incidence is highly affected by the conditions into which the mosquitoes are released. Compared to traditional vector control methods, Wolbachia-based approaches are much more environmentally friendly and can be effective in the medium/long term. On the flip side, they are much more complex and cost-intensive operations, requiring a substantial investment, infrastructure, trained personnel, coordination between agencies, and community engagement. Finally, we discuss recent evidence suggesting that the release of Wolbachia-transinfected mosquitoes has a moderate potential risk of spreading potentially dangerous genes in the environment.
Evaluation of Wolbachia infection in Aedes aegypti suggests low prevalence and highly heterogeneous distribution in Medellín, Colombia
32509rley Calle-Tobón, Raúl Rojo-Ospina, et al., Acta Tropica, 260. 2024-10-08 09:19:14.
Dengue virus, transmitted mainly by Aedes aegypti mosquitoes, is a significant public health challenge in tropical and subtropical countries, with an incidence that is growing at an alarming rate. The release of Wolbachia-carrying mosquitoes has been suggested as a strategy to reduce the incidence of multiple arboviruses. In Medellín, Colombia, large-scale releases of Wolbachia-infected Ae. aegypti mosquitoes were performed between 2017 and 2022 by the World Mosquito Program to facilitate population replacement. In this study, we evaluated the prevalence and distribution of Wolbachia-infected Ae. aegypti two years after completion of these releases. We conducted the sampling across 19 communes in Medellín, using 416 ovitraps to collect Ae. aegypti eggs from epidemiological weeks 26 to 41 in 2023. Upon hatching the collected eggs, we identified and pooled adult female Ae. aegypti for DNA extraction. Subsequently, we conducted PCR assays for the detection of Wolbachia infection in these mosquitoes. We used maximum likelihood estimation (MLE) and Bayesian methods to estimate the prevalence of Wolbachia infection, while using QGIS to analyze spatial distribution of infection in the region. A total of 774 female Ae. aegypti mosquitoes from 182 pools were evaluated. We detected Wolbachia in 33.5 % of pools, with an estimated individual minimum infection rate of 9.5 % and a maximum of 33.2 %. The prevalence varied significantly across communes, with the highest rates observed in the northeastern and southwestern areas. Spatial analysis revealed a highly heterogeneous island-like distribution of Wolbachia across Medellín with a few hotspots. The observed Wolbachia prevalence in this work was lower than previously reported. We suspect a decline in the prevalence of Wolbachia-infected Ae. aegypti mosquitoes in Medellín following the completion of their release.
Current status of the sterile insect technique for the suppression of mosquito populations on a global scale
32019Bouyer, J., Infectious Diseases of Poverty, 13. 2024-10-01 16:03:32.
The World Health Organization (WHO) has emphasized the urgent need for alternative strategies to chemical insecticides for controlling mosquito populations, particularly the invasive Aedes species, which are known vectors of arboviruses. Among these alternative approaches, the sterile insect technique (SIT) is experiencing rapid development, with numerous pilot trials being conducted worldwide. This review aims to elucidate the principles of SIT and highlight the significant recent advancements that have facilitated its scalability. I also employ a phased conditional approach to categorize the progression of 39 projects, drawing on peer reviewed studies, press releases and direct communication with project managers. This review indicates that a substantial number of projects illustrate the efficacy of SIT in suppressing Aedes populations, with one project even demonstrating a reduction in dengue incidence. I offer several recommendations to mitigate potential failures and address the challenges of compensation and overcompensation when implementing SIT field trials. Furthermore, I examine the potential implications of male mating harassment on the effectiveness of SIT in reducing disease transmission. This comprehensive assessment underscores the promise of SIT as a viable strategy for mosquito control. The insights gained from these trials not only contribute to the understanding of SIT’s effectiveness but also highlight the importance of careful project management and ecological considerations in the pursuit of public health objectives.
Mark–Release–Recapture Trial with Aedes albopictus (Diptera, Culicidae) Irradiated Males: Population Parameters and Climatic Factors
31590Amaro FIF, Soares P, Velo E, et al, Insects, 15. 2024-09-17 12:23:28.
Aedes albopictus mosquitoes spread arboviruses like dengue, Zika, or chikungunya. The Sterile Insect Technique (SIT) can be used as a prevention and control tool against Ae. albopictus populations. Mark–release–recapture (MRR) trials are fundamental to estimate the size of the wild population in target areas and to determine the dispersal capacity and survival of sterile males in open field conditions. Environmental conditions can affect the MRR trial’s results; thus, the influence of climatic factors on the first MRR trial with Ae. albopictus marked sterile males conducted in Portugal was analyzed. In October 2022, for three consecutive weeks at two different release points, 84,000 sterile males were released over 50 ha of the study area, in the municipality of Faro, Southern Portugal, and mosquitoes were recaptured by human landing collection (HLC) one, two, four, and six days after release. Distance travelled, daily survival, and life expectancy in the field were estimated for the released sterile males and the influence of climatic factors was analyzed. Despite no statistically significant association being found with humidity, temperature, and precipitation, it is crucial to plan MRRs, considering weather conditions for a more efficient application of the SIT in an integrated vector management program.
Bunyamwera Virus Infection of Wolbachia-Carrying Aedes aegypti Mosquitoes Reduces Wolbachia Density
31318Lefteri, D.A.; Rainey, S.M.; Murdochy, S.M.; Sinkins, S.P., Viruses, 16. 2024-09-03 15:42:46.
Wolbachia symbionts introduced into Aedes mosquitoes provide a highly effective dengue virus transmission control strategy, increasingly utilised in many countries in an attempt to reduce disease burden. Whilst highly effective against dengue and other positive-sense RNA viruses, it remains unclear how effective Wolbachia is against negative-sense RNA viruses. Therefore, the effect of Wolbachia on Bunyamwera virus (BUNV) infection in Aedes aegypti was investigated using wMel and wAlbB, two strains currently used in Wolbachia releases for dengue control, as well as wAu, a strain that typically persists at a high density and is an extremely efficient blocker of positive-sense viruses. Wolbachia was found to reduce BUNV infection in vitro but not in vivo. Instead, BUNV caused significant impacts on density of all three Wolbachia strains following infection of Ae. aegypti mosquitoes. The ability of Wolbachia to successfully persist within the mosquito and block virus transmission is partially dependent on its intracellular density. However, reduction in Wolbachia density was not observed in offspring of infected mothers. This could be due in part to a lack of transovarial transmission of BUNV observed. The results highlight the importance of understanding the complex interactions between multiple arboviruses, mosquitoes and Wolbachia in natural environments, the impact this can have on maintaining protection against diseases, and the necessity for monitoring Wolbachia prevalence at release sites.
Gene Drives: A Powerful and Controversial Genetic Technology
31456Biomedical and Life Sciences Simplified, YouTube, 2024-09-03 09:01:56.
Gene drives represent a powerful yet controversial genetic technique that challenges traditional Mendelian inheritance. By introducing self-propagating changes into an organism's genome, gene drives can ensure that a specific mutation becomes homozygous in offspring, effectively spreading it through a population. This powerful tool has potential applications in eradicating mosquito-borne diseases like malaria and Zika, combating drug-resistant pathogens, and controlling invasive species. But how exactly does it work? This video breaks down the science behind gene drives and the role of CRISPR technology.
Wolbachia endosymbionts in Drosophila regulate the resistance to Zika virus infection in a sex dependent manner
30938Ghada Tafesh-Edwards, Margarita Kyza Karaviot, Klea Markollari, et al, Frontiers in Microbiology, 15. 2024-07-24 16:02:48.
Drosophila melanogaster has been used extensively for dissecting the genetic and functional bases of host innate antiviral immunity and virus-induced pathology. Previous studies have shown that the presence of Wolbachia endosymbionts in D. melanogaster confers resistance to infection by certain viral pathogens. Zika virus is an important vector-borne pathogen that has recently expanded its range due to the wide geographical distribution of the mosquito vector. Here, we describe the effect of Wolbachia on the immune response of D. melanogaster adult flies following Zika virus infection. First, we show that the presence of Wolbachia endosymbionts promotes the longevity of uninfected D. melanogaster wild type adults and increases the survival response of flies following Zika virus injection. We find that the latter effect is more pronounced in females rather than in males. Then, we show that the presence of Wolbachia regulates Zika virus replication during Zika virus infection of female flies. In addition, we demonstrate that the antimicrobial peptide-encoding gene Drosocin and the sole Jun N-terminal kinase-specific MAPK phosphatase Puckered are upregulated in female adult flies, whereas the immune and stress response gene TotM is upregulated in male individuals. Finally, we find that the activity of RNA interference and Toll signaling remain unaffected in Zika virus-infected female and male adults containing Wolbachia compared to flies lacking the endosymbionts. Our results reveal that Wolbachia endosymbionts in D. melanogaster affect innate immune signaling activity in a sex-specific manner, which in turn influences host resistance to Zika virus infection. This information contributes to a better understanding of the complex interrelationship between insects, their endosymbiotic bacteria, and viral infection. Interpreting these processes will help us design more effective approaches for controlling insect vectors of infectious disease.
Culex-Transmitted Diseases: Mechanisms, Impact, and Future Control Strategies using Wolbachia
31030Madhav, M.; Blasdell, K.R.; Trewin, B.; Paradkar, P.N.; López-Denman, A.J., Viruses, 16:1134. 2024-07-15 11:40:32.
Mosquitoes of the Culex genus are responsible for a large burden of zoonotic virus transmission globally. Collectively, they play a significant role in the transmission of medically significant diseases such as Japanese encephalitis virus and West Nile virus. Climate change, global trade, habitat transformation and increased urbanisation are leading to the establishment of Culex mosquitoes in new geographical regions. These novel mosquito incursions are intensifying concerns about the emergence of Culex-transmitted diseases and outbreaks in previously unaffected areas. New mosquito control methods are currently being developed and deployed globally. Understanding the complex interaction between pathogens and mosquitoes is essential for developing new control strategies for Culex species mosquitoes. This article reviews the role of Culex mosquitos as vectors of zoonotic disease, discussing the transmission of viruses across different species, and the potential use of Wolbachia technologies to control disease spread. By leveraging the insights gained from recent successful field trials of Wolbachia against Aedes-borne diseases, we comprehensively discuss the feasibility of using this technique to control Culex mosquitoes and the potential for the development of next generational Wolbachia-based control methods.
Scientists use gut bacteria to prevent mosquito-borne diseases
29900CGTN, 2024-05-07 17:04:49.
Chinese scientists have developed a more natural strategy to prevent mosquito-borne diseases by changing insects' gut microbes, which might be used as an alternative to controversial experiments that see genetically-modified mosquitoes released in Florida. Mosquito-borne viruses, such as dengue and Zika, cause several potentially fatal human viral infections. Dengue viruses infect approximately 390 million each year globally. An epidemic survey over the past decade documented frequent dengue outbreaks in Xishuangbanna and Lincang, both in southwest China's Yunnan Province. But few have been reported in neighboring cities of Wenshan and Pu'er. The marked different prevalence stimulated the curiosity of researchers from Tsinghua University and the Yunnan Academy of Animal Science and Veterinary Sciences.
Bacteria found in mosquito guts could help scientists fight dengue, Zika
29548Catherine Offord, Science, 2024-04-25 17:12:41.
A team in China probing the guts of local mosquitoes has found a potential helper in the fight against two human diseases. Researchers identified a new bacterium that disables the viruses responsible for dengue and Zika before they can establish an infection in the insects. Although early stage, the work, reported this week in Science, paves the way for studying the bacterium’s effect on disease transmission in the real world. It wouldn’t be the first time a microbe is used to thwart mosquito-borne diseases. About 15 years ago, researchers discovered that a different bacterium, Wolbachia, reduces the insects’ ability to transmit dengue, among other viruses. Following successful field trials, Wolbachia is now used to help control dengue in more than a dozen countries. But an extra weapon to help control mosquito-borne diseases is welcome—especially as the insects become resistant to current insecticides. The new study is “promising,” says Nsa Dada, a vector biologist at Arizona State University who was not involved in the work. “It’s important that we explore … new tools that can complement existing control measures.” Lacking proven treatments or widely available vaccines, dengue is the most common mosquito-borne viral disease worldwide, infecting some 400 million people each year. Although most cases are mild or asymptomatic, about one in 20 people who get sick develop severe dengue, which can be fatal.
The Perpetual Vector Mosquito Threat and Its Eco-Friendly Nemeses
29088Miranda, L.S.; Rudd, S.R.; Mena, O.; Hudspeth, P.E.; Barboza-Corona, J.E.; Park, H.-W.; Bideshi, D.K., Biology, 13:182. 2024-04-04 09:42:02.
Aedes, Culex, and Anopheles mosquitoes are the most prolific arthropod vectors of viral and parasitic agents of debilitating and lethal diseases in humans and animals. Despite some success in integrated pest management programs to control vectors, mosquito-borne diseases, such as dengue and dengue hemorrhagic fever, yellow fever, chikungunya, West Nile, and Zika, and parasitic diseases, such as malaria, lymphatic filariasis, and river blindness, continue to threaten the health and well-being of half the world’s population, many of whom live in economically and medically challenged societies. The perpetual problem inflicted by vector-borne diseases is compounded by the selection for resistance to synthetic pesticides, globalization, and climate change. The latter appears to be the most significant factor implicated in the geographic expansion of mosquitoes. Here, we present a review of these challenges and highlight traditional vector control strategies that employ synthetic pesticides, and “green” eco-friendly technologies that include SIT, IIT, RIDL, CRISPR/Cas9/Cas13 gene drive systems, and biological control, with an emphasis on Lysinibacillus sphaericus and Bacillus thuringiensis subsp. israelensis (Bti).
Wolbachia Infection through Hybridization to Enhance an Incompatible Insect Technique-Based Suppression of Aedes albopictus in Eastern Spain
29086Cholvi M, Trelis M, Bueno-Marí R, Khoubbane M, Gil R, Marcilla A, Moretti R., Insects, 15. 2024-04-04 09:29:30.
Wolbachia bacteria occur naturally as symbionts of many insect species and are responsible for various phenomena that modify the hosts’ reproductive biology. Among them, cytoplasmic incompatibility (CI) refers to the sterility of eggs produced by crosses between infected males and females that are uninfected or infected by a non-compatible strain of these bacteria. CI can be exploited for vector control by establishing an opportune Wolbachia infection in a laboratory population of a target insect species and then releasing the infected males into the environment as sterilizing agents. In the present work, a suitable Wolbachia strain was introduced into a Spanish population of the Asian tiger mosquito, Aedes albopictus, through hybridization with the laboratory line, ARwP, already tested as an efficient control tool against this vector. The obtained hybrids were compared with the ARwP to ascertain the effects derived from transferring the infection to a different Ae. albopictus population. No significant differences between lines were found regarding survival, female fecundity, and egg fertility. Importantly, the eggs produced by crosses between males of the hybrid lines and unmodified wild females were 99.9% sterile. This result encourages further studies to explore the feasibility of a Wolbachia-based control program against the Asian tiger mosquito in Spain.
Detection and quantification of natural Wolbachia in Aedes aegypti in Metropolitan Manila, Philippines using locally designed primers
29084Reyes JIL, Suzuki T, Suzuki Y, Watanabe K., Frontiers in Cellular and Infection Microbiology, 2024-04-04 09:23:33.
The Philippines bears health and economic burden caused by high dengue cases annually. Presently, the Philippines still lack an effective and sustainable vector management. The use of Wolbachia, a maternally transmitted bacterium, that mitigate arbovirus transmission has been recommended. Cytoplasmic incompatibility and viral blocking, two characteristics that make Wolbachia suitable for vector control, depend on infection prevalence and density. There are no current Wolbachia release programs in the Philippines, and studies regarding the safety of this intervention. Here, we screened for Wolbachia in Aedes aegypti collected from Metropolitan Manila, Philippines. We designed location-specific primers for qPCR to test whether this improved Wolbachia detection in Ae. aegypti. We explored if host sex and Wolbachia strain could be potential factors affecting Wolbachia density.
Intra-lineage microevolution of Wolbachia leads to the emergence of new cytoplasmic incompatibility patterns
28889Namias A, Ngaku A, Makoundou P, Unal S, Sicard M, Weill M, PLoS Biology, 2024-02-13 20:25:15.
Mosquitoes of the Culex pipiens complex are worldwide vectors of arbovirus, filarial nematodes, and avian malaria agents. In these hosts, the endosymbiotic bacteria Wolbachia induce cytoplasmic incompatibility (CI), i.e., reduced embryo viability in so-called incompatible crosses. Wolbachia infecting Culex pipiens (wPip) cause CI patterns of unparalleled complexity, associated with the amplification and diversification of cidA and cidB genes, with up to 6 different gene copies described in a single wPip genome. In wPip, CI is thought to function as a toxin-antidote (TA) system where compatibility relies on having the right antidotes (CidA) in the female to bind and neutralize the male’s toxins (CidB). By repeating crosses between Culex isofemale lines over a 17 years period, we documented the emergence of a new compatibility type in real time and linked it to a change in cid genes genotype. We showed that loss of specific cidA gene copies in some wPip genomes results in a loss of compatibility. More precisely, we found that this lost antidote had an original sequence at its binding interface, corresponding to the original sequence at the toxin’s binding interface. We showed that these original cid variants are recombinant, supporting a role for recombination rather than point mutations in rapid CI evolution. These results strongly support the TA model in natura, adding to all previous data acquired with transgenes expression.
Maxizyme-mediated suppression of chikungunya virus replication and transmission in transgenic Aedes aegypti mosquitoes
28819Mishra P, Balaraman V, Fraser Jr. M, Frontiers in Microbiology, 14. 2024-01-23 14:40:52.
Chikungunya virus (CHIKV) is an emerging mosquito-borne pathogen of significant public health importance. There are currently no prophylactic vaccines or therapeutics available to control CHIKV. One approach to arbovirus control that has been proposed is the replacement of transmission-competent mosquitoes with those that are refractory to virus infection. Several transgene effectors are being examined as potentially useful for this population replacement approach. We previously demonstrated the successful use of hammerhead ribozymes (hRzs) as an antiviral effector transgene to control CHIKV infection of, and transmission by, Aedes mosquitoes. In this report we examine a maxizyme approach to enhance the catalytic activity and prevent virus mutants from escaping these ribozymes. We designed a maxizyme containing minimized (monomer) versions of two hRzs we previously demonstrated to be the most effective in CHIKV suppression. Three versions of CHIKV maxizyme were designed: Active (Mz), inactive (ΔMz), and a connected CHIKV maxizyme (cMz). The maxizymes with their expression units (Ae-tRNA val promoter and its termination signal) were incorporated into lentivirus vectors with selection and visualization markers. Following transformation, selection, and single-cell sorting of Vero cells, clonal cell populations were infected with CHIKV at 0.05 and 0.5 MOI, and virus suppression was assessed using TCID50-IFA, RT-qPCR, and caspase-3 assays. Five transgenic mosquito lines expressing cMz were generated and transgene insertion sites were confirmed by splinkerette PCR. Our results demonstrate that Vero cell clones expressing Mz exhibited complete inhibition of CHIKV replication compared to their respective inactive control version or the two parent hRzs. Upon oral challenge of transgenic mosquitoes with CHIKV, three out of the five lines were completely refractory to CHIKV infection, and all five lines tested negative for salivary transmission. Altogether, this study demonstrates that maxizymes can provide a higher catalytic activity and viral suppression than hRzs.
Wolbachia still works when it is warm
28809McKay, A., Nature Ecology and Evolution, 8. 2024-01-16 13:59:40.
Wolbachia is a maternally inherited endosymbiotic bacterium that can impede the transmission of viruses such as dengue and Zika by some mosquito vectors to humans. Over the past decade, this self-sustaining disease-control method has been rolled out in cities of increasing size; 2023 saw the largest demonstration of efficacy to date, in Medellín, Colombia. However, two key biological mechanisms that enable Wolbachia-carrying mosquitoes to spread in a population — cytoplasmic incompatibility and maternal transmission — have been shown in laboratory conditions to decline under high temperatures. Writing in Nature Climate Change in August 2023, Vásquez and colleagues conducted computational simulations to explore whether near-future warming scenarios are likely to affect the efficacy of Wolbachia-based biocontrol. The models combine empirical estimates of thermal sensitivity for one strain of Wolbachia with projections of future average temperatures and heatwaves for Cairns, Australia and Nha Trang City, Vietnam, two locations where the intervention has been deployed. Estimates of the bacterium’s successful persistence in the mosquito population remain high under tested projections of moderate and severe average warming to 2050. However, the simulations also show reduced efficacy under more variable thermal extremes; these interventions have the potential to fail under extreme warming with long heatwave durations. We selected this paper for our Year in Review collection because it exemplifies how insect thermal biology and global change analysis can offer insights relevant to human health. Testing whether climate change influences Wolbachia-based biocontrol helps to illuminate whether the promising technology is likely to remain feasible in coming years.
Antiviral Wolbachia strains associate with Aedes aegypti endoplasmic reticulum membranes and induce lipid droplet formation to restrict dengue virus replication
28757Robson K. Loterio, Ebony A. Monson, Rachel Templin, Jyotika T. de Bruyne, Heather A. Flores, Jason M. Mackenzie, Georg Ramm, Karla J. Helbig, Cameron P. Simmons, Johanna E. Fraser, Applied and Environmental Microbiology, 2023-12-22 16:16:14.
Wolbachia are a genus of insect endosymbiotic bacteria which includes strains wMel and wAlbB that are being utilized as a biocontrol tool to reduce the incidence of Aedes aegypti-transmitted viral diseases like dengue. However, the precise mechanisms underpinning the antiviral activity of these Wolbachia strains are not well defined. Here, we generated a panel of Ae. aegypti-derived cell lines infected with antiviral strains wMel and wAlbB or the non-antiviral Wolbachia strain wPip to understand host cell morphological changes specifically induced by antiviral strains. Antiviral strains were frequently found to be entirely wrapped by the host endoplasmic reticulum (ER) membrane, while wPip bacteria clustered separately in the host cell cytoplasm. ER-derived lipid droplets (LDs) increased in volume in wMel- and wAlbB-infected cell lines and mosquito tissues compared to cells infected with wPip or Wolbachia-free controls. Inhibition of fatty acid synthase (required for triacylglycerol biosynthesis) reduced LD formation and significantly restored ER-associated dengue virus replication in cells occupied by wMel. Together, this suggests that antiviral Wolbachia strains may specifically alter the lipid composition of the ER to preclude the establishment of dengue virus (DENV) replication complexes. Defining Wolbachia’s antiviral mechanisms will support the application and longevity of this effective biocontrol tool that is already being used at scale.
Viruses that ‘infect’ viruses: Cas12f1 and Cas9 gene drive in HSV1
28673Hongsheng Dai, Qiaorui Yao, Zhuangjie Lin, Keyuan Lai, Xianyin Zeng, Guangxiong Lei, Tongwen Zhang, bioRxiv, 2023-12-06 12:08:32.
Cas9-based synthetic gene drives constitute some minimal elements capable of editing DNA with sequence specificity. However, they face high resistance rate and mitigation strategies developed so far are difficult to implement. Here, we engineered herpes simplex virus type 1 (HSV1) as a vector for gene drive, generated viruses carrying either Cas9 or Cas12f1 gene drives, and compared their performance in spreading the gene drive and causing resistance. Based on phenotypical staining and NGS sequencing, we showed that HSV1 supported fast and effective transmission of gene drive, and Cas12f1 gene drive yielded higher conversion and lower resistance than Cas9 gene drive. Cas12f1 gene drive is a promising alternative to Cas9 gene drive, and HSV1 is a reliable and fast platform to evaluate gene drive. Gene drive viruses worked like pathogens that specifically infect viruses and could potentially be used to attenuate viral infection.
Male-killing virus leads to more female moths
28365Anonymous, Nature, 2023-11-07 09:51:08.
Keisuke Nagamine at Minami Kyushu University in Miyazaki, Japan, and his colleagues have identified another virus that kills male embryos of the tobacco caterpillar, Spodoptera litura. Female moths infected with the virus produced an equal number of male and female embryos, but almost no male embryos survived. The virus doesn’t share male-killing genes with other known male-killing viruses or bacteria, suggesting that these mechanisms evolved independently.
Male-killing virus in a noctuid moth Spodoptera litura
28367K. Nagamine, Y. Kanno, K. Sahara, T. Fujimoto, A. Yoshido, Y. Ishikawa, M. Terao, D. Kageyama and Y. Shintani, Proceedings of the National Academy of Sciences, 120:e2312124120. 2023-11-06 09:56:12.
A female-biased sex ratio is considered advantageous for the cytoplasmic elements that inhabit sexually reproducing organisms. There are numerous examples of bacterial symbionts in the arthropod cytoplasm that bias the host sex ratio toward females through various means, including feminization and male killing. Recently, maternally inherited RNA viruses belonging to the family Partitiviridae were found to cause male killing in moths and flies, but it was unknown whether male-killing viruses were restricted to Partitiviridae or could be found in other taxa. Here, we provide compelling evidence that a maternally inherited RNA virus, Spodoptera litura male-killing virus (SlMKV), selectively kills male embryos of the tobacco caterpillar Spodoptera litura, resulting in all-female broods. SlMKV injected into uninfected S. litura can also be inherited maternally and causes male killing. SlMKV has five genomic segments encoding seven open reading frames, has no homolog of known male-killing genes, and belongs to an unclassified group of arthropod-specific viruses closely related to Tolivirales. When transinfected into larvae, both male and female recipients allow SlMKV to proliferate, but only males die at the pupal stage. The viral RNA levels in embryonic and pupal male killing suggest that the mechanism of male killing involves the constitutive expression of viral products that are specifically lethal to males, rather than the male-specific expression of viral products. Our results, together with recent findings on male-killing partiti-like viruses, suggest that diverse viruses in arthropods tend to acquire male killing independently and that such viruses may be important components of intragenomic conflict in arthropods.
Wolbachia interferes with Zika virus replication by hijacking cholesterol metabolism in mosquito cells
28094B. Edwards, E. A.-O. X. Ghedin and D. A.-O. Voronin, Microbiology Spectrum, 2023-10-09 07:37:16.
Zika virus is a member of the arbovirus Flaviviridae family transmitted by Aedes mosquitos and it is associated with microcephaly in infants born to infected mothers. Wolbachia is an intracellular gram-negative alpha-proteobacteria that infects many species of arthropods, including mosquitos. The presence of Wolbachia in mosquitos has been shown to control the vector population and suppress arbovirus transmission. One mechanism of Wolbachia-mediated interference with virus replication is competition over host resources between Wolbachia and the virus. We hypothesize that cholesterol metabolism is involved in Wolbachia-mediated virus suppression due to its important role in Zika virus replication. In this study, we determined that Wolbachia impacted virus replication by altering cholesterol biosynthesis in Aedes albopictus C6/36 cells, diverting resources from the host cell mevalonate (MVA) pathway to fulfill the needs of the bacteria. This resulted in a decrease of total cholesterol, increased Wolbachia loads, and decreased viral titers. Inhibition of the MVA pathway using fluvastatin decreased total cholesterol and viral titers, mimicking the effects of Wolbachia on the virus in Wolbachia-free cells. We also found that Wolbachia-infected cells had depleted lipid droplets, the main component of which is cholesterol esters. We confirmed that cholesterol esterases were upregulated in response to virus infection in C6/36 cells. Functional analysis showed that alteration of cholesterol metabolism simulated Wolbachia-mediated inhibition of virus infection in C6/36 cells. Our study provides a mechanism behind Wolbachia-induced interference of arbovirus replication and could help advance strategies to control arbovirus pathogens in insect vectors and human infections. IMPORTANCE Arthropod-borne viruses are emerging pathogens that are spread widely by mosquitos. Zika virus is an arbovirus that can infect humans and be transmitted from an infected mother to the fetus, potentially leading to microcephaly in infants. One promising strategy to prevent disease caused by arboviruses is to target the insect vector population. Recent field studies have shown that mosquito populations infected with Wolbachia bacteria suppress arbovirus replication and transmission. Here, we describe how intracellular bacteria redirect resources within their host cells and suppress Zika virus replication at the cellular level. Understanding the mechanism behind Wolbachia-induced interference of arbovirus replication could help advance strategies to control arbovirus pathogens in insect vectors and human populations. FAU - Edwards, Brent
Jamestown Canyon virus is transmissible by Aedes aegypti and is only moderately blocked by Wolbachia co-infection
27755M. J. Lau, H. L. C. Dutra, M. J. Jones, B. P. McNulty, A. M. Diaz, F. Ware-Gilmore and E. A. McGraw, PLOS Neglected Tropical Diseases, 17. 2023-09-05 07:50:16.
Jamestown Canyon virus (JCV), a negative-sense arbovirus, is increasingly common in the upper Midwest of the USA. Transmitted by a range of mosquito genera, JCV's primary amplifying host is white-tailed deer. Aedes aegypti is responsible for transmitting various positive-sense viruses globally including dengue (DENV), Zika, chikungunya, and Yellow Fever. Ae. aegypti's distribution, once confined to the tropics, is expanding, in part due to climate change. Wolbachia, an insect endosymbiont, limits the replication of co-infecting viruses inside insects. The release and spread of the symbiont into Ae. aegypti populations have been effective in reducing transmission of DENV to humans, although the mechanism of Wolbachia-mediated viral blocking is still poorly understood. Here we explored JCV infection potential in Ae. aegypti, the nature of the vector's immune response, and interactions with Wolbachia infection. We show that Ae. aegypti is highly competent for JCV, which grows to high loads and rapidly reaches the saliva after an infectious blood meal. The mosquito immune system responds with strong induction of RNAi and JAK/STAT. Neither the direct effect of viral infection nor the energetic investment in immunity appears to affect mosquito longevity. Wolbachia infection blocked JCV only in the early stages of infection. Wolbachia-induced immunity was small compared to that of JCV, suggesting innate immune priming does not likely explain blocking. We propose two models to explain why Wolbachia's blocking of negative-sense viruses like JCV may be less than that of positive-sense viruses, relating to the slowdown of host protein synthesis and the triggering of interferon-like factors like Vago. In conclusion, we highlight the risk for increased human disease with the predicted future overlap of Ae. aegypti and JCV ranges. We suggest that with moderate Wolbachia-mediated blocking and distinct biology, negative-sense viruses represent a fruitful comparator model to other viruses for understanding blocking mechanisms in mosquitoes.
wMel Wolbachia alters female post-mating behaviors and physiology in the dengue vector mosquito Aedes aegypti
27649J. Osorio, S. Villa-Arias, C. Camargo, L. F. Ramírez-Sánchez, L. M. Barrientos, C. Bedoya, G. Rúa-Uribe, S. Dorus, C. Alfonso-Parra and F. W. Avila, Communications Biology, 6:865. 2023-08-21 06:53:59.
Globally invasive Aedes aegypti disseminate numerous arboviruses that impact human health. One promising method to control Ae. aegypti populations is transinfection with Wolbachia pipientis, which naturally infects ~40–52% of insects but not Ae. aegypti. Transinfection of Ae. aegypti with the wMel Wolbachia strain induces cytoplasmic incompatibility (CI), allows infected individuals to invade native populations, and inhibits transmission of medically relevant arboviruses by females. Female insects undergo post-mating physiological and behavioral changes—referred to as the female post-mating response (PMR)—required for optimal fertility. PMRs are typically elicited by male seminal fluid proteins (SFPs) transferred with sperm during mating but can be modified by other factors, including microbiome composition. Wolbachia has modest effects on Ae. aegypti fertility, but its influence on other PMRs is unknown. Here, we show that Wolbachia influences female fecundity, fertility, and re-mating incidence and significantly extends the longevity of virgin females. Using proteomic methods to examine the seminal proteome of infected males, we found that Wolbachia moderately affects SFP composition. However, we identified 125 paternally transferred Wolbachia proteins, but the CI factor proteins (Cifs) were not among them. Our findings indicate that Wolbachia infection of Ae. aegypti alters female PMRs, potentially influencing control programs that utilize Wolbachia-infected individuals.
Wolbachia -mediated resistance to Zika virus infection in Aedes aegypti is dominated by diverse transcriptional regulation and weak evolutionary pressures
26604E. C. Boehm, A. S. Jaeger, H. J. Ries, D. Castañeda, A. M. Weiler, C. C. Valencia, J. Weger-Lucarelli, G. D. Ebel, S. L. O'Connor, T. C. Friedrich, M. Zamanian and M. T. Aliota, bioRxiv, 2023-06-26 13:09:35.
A promising candidate for arbovirus control and prevention relies on replacing arbovirus-susceptible Aedes aegypti populations with mosquitoes that have been colonized by the intracellular bacterium Wolbachia and thus have a reduced capacity to transmit arboviruses. This reduced capacity to transmit arboviruses is mediated through a phenomenon referred to as pathogen blocking. Pathogen blocking has primarily been proposed as a tool to control dengue virus (DENV) transmission, however it works against a range of viruses, including Zika virus (ZIKV). Despite years of research, the molecular mechanisms underlying pathogen blocking still need to be better understood. Here, we used RNA-seq to characterize mosquito gene transcription dynamics in Ae. aegypti infected with the w Mel strain of Wolbachia that are being released by the World Mosquito Program in Medellín, Colombia. Comparative analyses using ZIKV-infected, uninfected tissues, and mosquitoes without Wolbachia revealed that the influence of w Mel on mosquito gene transcription is multifactorial. Importantly, because Wolbachia limits, but does not completely prevent, replication of ZIKV and other viruses in coinfected mosquitoes, there is a possibility that these viruses could evolve resistance to pathogen blocking. Therefore, to understand the influence of Wolbachia on within-host ZIKV evolution, we characterized the genetic diversity of molecularly barcoded ZIKV virus populations replicating in Wolbachia -infected mosquitoes and found that within-host ZIKV evolution was subject to weak purifying selection and, unexpectedly, loose anatomical bottlenecks in the presence and absence of Wolbachia . Together, these findings suggest that there is no clear transcriptional profile associated with Wolbachia -mediated ZIKV restriction, and that there is no evidence for ZIKV escape from this restriction in our system. AUTHOR SUMMARY: When Wolbachia bacteria infect Aedes aegypti mosquitoes, they dramatically reduce the mosquitoes' susceptibility to infection with a range of arthropod-borne viruses, including Zika virus (ZIKV). Although this pathogen-blocking effect has been widely recognized, its mechanisms remain unclear. Furthermore, because Wolbachia limits, but does not completely prevent, replication of ZIKV and other viruses in coinfected mosquitoes, there is a possibility that these viruses could evolve resistance to Wolbachia -mediated blocking. Here, we use host transcriptomics and viral genome sequencing to examine the mechanisms of ZIKV pathogen blocking by Wolbachia and viral evolutionary dynamics in Ae. aegypti mosquitoes. We find complex transcriptome patterns that do not suggest a single clear mechanism for pathogen blocking. We also find no evidence that Wolbachia exerts detectable selective pressures on ZIKV in coinfected mosquitoes. Together our data suggest that it may be difficult for ZIKV to evolve Wolbachia resistance, perhaps due to the complexity of the pathogen blockade mechanism.
Wolbachia -induced inhibition of O’nyong nyong virus in Anopheles mosquitoes is mediated by Toll signaling and modulated by cholesterol
26465S. Pujhari, G. L. Hughes, N. Pakpour, Y. Suzuki and J. L. Rasgon, bioRxiv, 10.1101/2023.05.31.543096. 2023-06-01 07:41:43.
Enhanced host immunity and competition for metabolic resources are two main competing hypotheses for the mechanism of Wolbachia -mediated pathogen inhibition in arthropods. Using an Anopheles mosquito - somatic Wolbachia infection - O'nyong nyong virus (ONNV) model, we demonstrate that the mechanism underpinning Wolbachia -mediated virus inhibition is up-regulation of the Toll innate immune pathway. However, the viral inhibitory properties of Wolbachia were abolished by cholesterol supplementation. This result was due to Wolbachia -dependent cholesterol-mediated suppression of Toll signaling rather than competition for cholesterol between Wolbachia and virus. The inhibitory effect of cholesterol was specific to Wolbachia -infected Anopheles mosquitoes and cells. These data indicate that both Wolbachia and cholesterol influence Toll immune signaling in Anopheles mosquitoes in a complex manner and provide a functional link between the host immunity and metabolic competition hypotheses for explaining Wolbachia -mediated pathogen interference in mosquitoes. In addition, these results provide a mechanistic understanding of the mode of action of Wolbachia -induced pathogen blocking in Anophelines, which is critical to evaluate the long-term efficacy of control strategies for malaria and Anopheles -transmitted arboviruses. HIGHLIGHTS: Wolbachia inhibits O'nyong nyong virus (ONNV) in Anopheles mosquitoes. Enhanced Toll signaling is responsible for Wolbachia -induced interference of ONNV. Cholesterol suppresses Toll signaling to modulate Wolbachia -induced ONNV interference.
Wolbachia protects Drosophila melanogaster against two naturally occurring and virulent viral pathogens
25211G. Bruner-Montero and F. M. Jiggins, Scientific Reports, 13:8518. 2023-05-25 13:38:46.
Wolbachia is a common endosymbiont that can protect insects against viral pathogens. However, whether the antiviral effects of Wolbachia have a significant effect on fitness remains unclear. We have investigated the interaction between Drosophila melanogaster, Wolbachia and two viruses that we recently isolated from wild flies, La Jolla virus (LJV; Iflaviridae) and Newfield virus (NFV; Permutotetraviridae). Flies infected with these viruses have increased mortality rates, and NFV partially sterilizes females. These effects on fitness were reduced in Wolbachia-infected flies, and this was associated with reduced viral titres. However, Wolbachia alone also reduces survival, and under our experimental conditions these costs of the symbiont can outweigh the benefits of antiviral protection. In contrast, protection against the sterilizing effect of NFV leads to a net benefit of Wolbachia infection after exposure to the virus. These results support the hypothesis that Wolbachia is an important defense against the natural pathogens of D. melanogaster. Furthermore, by reducing the cost of Wolbachia infection, the antiviral effects of Wolbachia may aid its invasion into populations and help explain why it is so common in nature.
Dengue Exposure and Wolbachia wMel Strain Affects the Fertility of Quiescent Eggs of Aedes aegypti
25102M. T. Petersen, D. Couto-Lima, G. A. Garcia, M. G. Pavan, M. R. David and R. Maciel-de-Freitas, Viruses, 15. 2023-04-12 10:20:54.
(1) Background: The deployment of the bacterium Wolbachia to reduce arbovirus transmission is ongoing in several countries worldwide. When Wolbachia-carrying Aedes aegypti are released and established in the field, females may feed on dengue-infected hosts. The effects of simultaneous exposure on life-history traits of Ae. aegypti to Wolbachia wMel strain and dengue-1 virus DENV-1 remain unclear. (2) Methods: We monitored 4 groups (mosquitoes with either DENV-1 or Wolbachia, coinfected with DENV-1 and Wolbachia, as well as negative controls) to estimate Ae. aegypti survival, oviposition success, fecundity, collapsing and fertility of quiescent eggs for 12 weeks. (3) Results: Neither DENV-1 nor Wolbachia had a significant impact on mosquito survival nor on mosquito fecundity, although the last parameter showed a tendency to decrease with ageing. There was a significant decrease in oviposition success in individuals carrying Wolbachia. Wolbachia infection and storage time significantly increased egg collapse parameter on the egg viability assay, while DENV-1 had a slight protective effect on the first four weeks of storage. (4) Conclusions: Despite limitations, our results contribute to better understanding of the tripartite interaction of virus, bacteria and mosquito that may take place in field conditions and aid in guaranteeing the Wolbachia strategy success.
Scientists disable protective gene in mosquitoes, making them susceptible to disease
24937Texas A&M University, Phys Org, 2023-03-14 10:09:07.
Immune pathways that protect mosquitoes from human pathogens, including West Nile, Zika and dengue viruses were disabled by Texas A&M AgriLife Research scientists. The research study, "RNA interference is essential to modulating the pathogenesis of mosquito-borne viruses in the yellow fever mosquito Aedes aegypti," published in the Proceedings of the National Academy of Sciences, showed that the mosquito vector species Aedes aegypti becomes acutely susceptible to disease when the protective immune pathway is disabled. This discovery provides new insight into the ecological and evolutionary interactions occurring between mosquitoes and the pathogens they transmit to humans and animals.
A male-killing gene encoded by a symbiotic virus of Drosophila
24838D. Kageyama, T. Harumoto, K. Nagamine, A. Fujiwara, T. N. Sugimoto, A. Jouraku, M. Tamura, T. K. Katoh and M. Watada, Nature Communications, 14:1357. 2023-03-13 08:49:35.
In most eukaryotes, biparentally inherited nuclear genomes and maternally inherited cytoplasmic genomes have different evolutionary interests. Strongly female-biased sex ratios that are repeatedly observed in various arthropods often result from the male-specific lethality (male-killing) induced by maternally inherited symbiotic bacteria such as Spiroplasma and Wolbachia. However, despite some plausible case reports wherein viruses are raised as male-killers, it is not well understood how viruses, having much smaller genomes than bacteria, are capable of inducing male-killing. Here we show that a maternally inherited double-stranded RNA (dsRNA) virus belonging to the family Partitiviridae (designated DbMKPV1) induces male-killing in Drosophila. DbMKPV1 localizes in the cytoplasm and possesses only four genes, i.e., one gene in each of the four genomic segments (dsRNA1−dsRNA4), in contrast to ca. 1000 or more genes possessed by Spiroplasma or Wolbachia. We also show that a protein (designated PVMKp1; 330 amino acids in size), encoded by a gene on the dsRNA4 segment, is necessary and sufficient for inducing male-killing. Our results imply that male-killing genes can be easily acquired by symbiotic viruses through reassortment and that symbiotic viruses are hidden players in arthropod evolution. We anticipate that host-manipulating genes possessed by symbiotic viruses can be utilized for controlling arthropods.
GeneConvene Global Collaborative Webinar Series | Emerging Gene Drive Systems 2023
24769David O'Brochta, 2023-02-22 15:50:48.
Gene drive systems are being engineered in the laboratory and in some cases shown to be effective at rapidly altering target-gene frequencies in experimental populations. Much of this foundational work has been conducted in insects in the laboratory. This webinar series will focus on emerging potential applications of gene drive technology in a wide variety of organisms. These webinars are intended to inform audiences of the rationale for these development efforts, the current state of research and development and outstanding challenges.
Engineered Antiviral Sensor Targets Infected Mosquitoes
24617E. Dalla Benetta, A. J. Lopez-Denman, H.-H. Li, R. A. Masri, D. J. Brogan, M. Bui, T. Yang, M. Li, M. Dunn, M. J. Klein, S. Jackson, K. Catalan, K. R. Blasdell, P. Tng, I. Antoshechkin, L. S. Alphey, P. N. Paradkar and O. Akbari, bioRxiv, 2023.01.27.525922. 2023-01-27 09:22:47.
Escalating vector disease burdens pose significant global health risks, so innovative tools for targeting mosquitoes are critical. We engineered an antiviral strategy termed REAPER (vRNA Expression Activates Poisonous Effector Ribonuclease) that leverages the programmable RNA-targeting capabilities of CRISPR Cas13 and its potent collateral activity. Akin to a stealthy Trojan Horse hiding in stealth awaiting the presence of its enemy, REAPER remains concealed within the mosquito until an infectious blood meal is up taken. Upon target viral RNA infection, REAPER activates, triggering programmed destruction of its target arbovirus such as chikungunya. Consequently, Cas13 mediated RNA targeting significantly reduces viral replication and its promiscuous collateral activity can even kill infected mosquitoes. This innovative REAPER technology adds to an arsenal of effective molecular genetic tools to combat mosquito virus transmission.Competing Interest StatementO.S.A is a founder of both Agragene, Inc. and Synvect, Inc. with equity interest. The terms of this arrangement have been reviewed and approved by the University of California, San Diego in accordance with its conflict of interest policies. L.A is an adviser to Synvect, Inc and Biocentis Ltd., with financial interest in each. All other authors declare no competing interests.
Wolbachia-Virus interactions and arbovirus control through population replacement in mosquitoes
23675T. H. Ant, M. V. Mancini, C. J. McNamara, S. M. Rainey and S. P. Sinkins, Pathogens and Global Health, 2022-10-07 08:27:10.
Following transfer into the primary arbovirus vector Aedes aegypti, several strains of the intracellular bacterium Wolbachia have been shown to inhibit the transmission of dengue, Zika, and chikungunya viruses, important human pathogens that cause significant morbidity and mortality worldwide. In addition to pathogen inhibition, many Wolbachia strains manipulate host reproduction, resulting in an invasive capacity of the bacterium in insect populations. This has led to the deployment of Wolbachia as a dengue control tool, and trials have reported significant reductions in transmission in release areas. Here, we discuss the possible mechanisms of Wolbachia-virus inhibition and the implications for long-term success of dengue control. We also consider the evidence presented in several reports that Wolbachia may cause an enhancement of replication of certain viruses under particular conditions, and conclude that these should not cause any concerns with respect to the application of Wolbachia to arbovirus control.
Externalities modulate the effectiveness of the Wolbachia release programme
23611E. E. Ooi and A. Wilder-Smith, The Lancet Infectious Diseases, 2022-09-28 09:16:47.
Despite the remarkable outcome in Yogyakarta, the wMel approach also has some challenges. In particular, the extent to which ecological, weather, and other external factors influence the dissemination and establishment of wMel in complex urban environments remains unclear. Whether the high and sustained penetration rate of wMel and the efficacy in preventing dengue that was observed in Yogyakarta are readily reproducible in other regions of the world that are vulnerable to repeated outbreaks of such viral diseases is also unclear. We therefore welcome the study into the real-world effectiveness of large-scale wMel release in the urban setting of Rio de Janeiro.3 The study assessed the rate of wMel introgression at the neighbourhood level and its effect on the localised incidence of dengue and chikungunya. Mosquitoes were released over a 2·5-year period starting in August, 2017.
Outbreaks of arboviruses, biotechnological innovations and vector control: facing the unexpected
23438C. Boëte, Innovative Strategies for Vector Control, 6:219-231. 2022-08-19 09:19:15.
Outbreaks of arboviruses have occurred in the last decades in many places around the world and a variety of responses have been taken in order to control them. Responses ranged from vaccination campaigns to the use of conventional vector control methods. Innovative approaches relying on biotechnological novelties, often still under development, have been considered despite the lack of solid evidence of their efficacy. While discussing these different aspects of the fight against vector-borne diseases with a focus on the context of outbreaks, this chapter considers the social and ethical aspects related to both the rhetoric and the discussion about the implementation of new and innovative approaches.
Changing mosquito genes, spreading bacteria: Science sees success vs dengue
23421C. E. Baclig, INQUIRER.NET, 2022-08-10 08:51:03.
Wolbachia, according to WMP, are extremely common bacteria that occur naturally in 50 percent of insect species, including mosquitoes, fruit flies, moths, dragonflies, and butterflies. Aedes aegypti or dengue-carrying mosquitoes, however, do not normally carry Wolbachia. Studies showed that the bacteria make it difficult for viruses, such as dengue, Zika, chikungunya, and yellow fever, to reproduce inside the Aedes aegypti mosquitoes—making the mosquitoes much less likely to spread viruses when they bite people. “This means that when Aedes aegypti mosquitoes carry natural Wolbachia bacteria, the transmission of viruses like dengue, Zika, chikungunya, and yellow fever is reduced,” the organization explained. The WMP breeds Wolbachia-carrying mosquitoes and releases them into areas hit by mosquito-borne diseases. This means there will be “less risk of disease in communities where Wolbachia is established in the local mosquito population.”
Wolbachia wPip Blocks Zika Virus Transovarial Transmission in Aedes albopictus
23333Y. Guo, J. Guo, Y. Li, X. Zheng and Y. Wu, Microbiol Spectrum, e0263321. 2022-07-27 08:36:55.
Area-wide application of Wolbachia to suppress mosquito populations and their transmitted viruses has achieved success in multiple countries. However, the mass release of Wolbachia-infected male mosquitoes involves a potential risk of accidentally releasing fertile females. In this study, we employed ovarian cells of the Ae. albopictus GUA, HC, and GT lines, which exhibit key traits, and compared them to better understand how Wolbachia inhibits ZIKV transovarial transmission. Our results showed an almost complete blockade of ZIKV transmission in HC female mosquitoes. Wolbachia in natively infected GUA mosquitoes negative affected ZIKV, and this interference was shown by slightly lower loads than those in HC mosquitoes. Overall, our work helps show how Wolbachia blocks ZIKV expansion and maintenance in the ovaries of Ae. albopictus and aids in understanding Wolbachia-ZIKV interactions in mosquitoes.
Intron-derived small RNAs for silencing viral RNAs in mosquito cells
23054P. Y. L. Tng, L. Z. Carabajal Paladino, M. A. E. Anderson, Z. N. Adelman, R. Fragkoudis, R. Noad and L. Alphey, PLOS Neglected Tropical Diseases, 16:e0010548. 2022-06-23 15:14:23.
Aedes aegypti and Ae. albopictus are the main vectors of mosquito-borne viruses of medical and veterinary significance. Many of these viruses have RNA genomes. Exogenously provided, e.g. transgene encoded, small RNAs could be used to inhibit virus replication, breaking the transmission cycle. We tested, in Ae. aegypti and Ae. albopictus cell lines, reporter based strategies for assessing the ability of two types of small RNAs to inhibit a chikungunya virus (CHIKV) derived target. Both types of small RNAs use a Drosophila melanogasterpremiRNA-1 based hairpin for their expression, either with perfect base-pairing in the stem region (shRNA-like) or containing two mismatches (miRNA-like). The pre-miRNA-1 stem loop structure was encoded within an intron; this allows co-expression of one or more proteins, e.g. a fluorescent protein marker tracking the temporal and spatial expression of the small RNAs in vivo. Three reporter-based systems were used to assess the relative silencing efficiency of ten shRNA-like siRNAs and corresponding miRNA-like designs. Two systems used a luciferase reporter RNA with CHIKV RNA inserted either in the coding sequence or within the 3’ UTR. A third reporter used a CHIKV derived split replication system. All three reporters demonstrated that while silencing could be achieved with both miRNA-like and shRNA-like designs, the latter were substantially more effective. Dcr-2 was required for the shRNA-like siRNAs as demonstrated by loss of inhibition of the reporters in Dcr-2 deficient cell lines. These positive results in cell culture are encouraging for the potential use of this pre-miRNA-1-based system in transgenic mosquitoes.
Modeling the impact of genetically modified male mosquitoes in the spatial population dynamics of Aedes aegypti
23212M. R. da Silva, P. H. G. Lugão, F. Prezoto and G. Chapiro, Scientific Reports, 12:9112. 2022-06-01 06:29:36.
The mosquito Aedes aegypti is the primary vector of diseases such as dengue, Zika, chikungunya, and yellow fever. Improving control techniques requires a better understanding of the mosquito’s life cycle, including spatial population dynamics in endemic regions. One of the most promising techniques consists of introducing genetically modified male mosquitoes. Several models proposed to describe this technique present mathematical issues or rely on numerous parameters, making their application challenging to real-world situations. We propose a model describing the spatial population dynamics of the Aedes aegypti in the presence of genetically modified males. This model presents some mathematical improvements compared to the literature allowing deeper mathematical analysis. Moreover, this model relies on few parameters, which we show how to obtain or estimate from the literature. Through numerical simulations, we investigate the impacts of environmental heterogeneity, the periodicity of genetically modified male releases, and released genetically modified males quantity on the population dynamics of Aedes aegypti. The main results point to that the successful application of this vector control technique relies on releasing more than a critical amount of modified males with a frequency exceeding a specific critical value.
Novel molecular approaches to combat vectors and vector-borne viruses: Special focus on RNA interference (RNAi) mechanisms
22657A. Agarwal, D. K. Sarma, D. Chaurasia and H. S. Maan, Acta Tropica, 2022-05-24 06:45:12.
Vector-borne diseases, such as dengue, chikungunya, zika, yellow fever etc pose significant burden among the infectious diseases globally, especially in tropical and sub-tropical regions. Globalization, deforestation, urbanization, climate change, uncontrolled population growth, inadequate waste management and poor vector-management infrastructure have all contributed to the expansion of vector habitats and subsequent increase in vector-borne diseases throughout the world. Conventional vector control methods, such as use of insecticides, have significant negative environmental repercussions in addition to developing resistance in vectors. Till date, a very few vaccines or antiviral therapies have been approved for the treatment of vector borne diseases. In this review, we have discussed emerging molecular approaches like CRISPR (clustered regularly interspaced short palindromic repeats)/Cas-9, sterile insect technique (SIT), release of insects carrying a dominant lethal (RIDL), Wolbachia (virus transmission blocking) and RNA interference (RNAi) to combat vector and vector-borne viruses. Due to the extensive advancements in RNAi research, a special focus has been given on its types, biogenesis, mechanism of action, delivery and experimental studies evaluating their application as anti-mosquito and anti-viral agent. These technologies appear to be highly promising in terms of contributing to vector control and antiviral drug development, and hence can be used to reduce global vector and vector-borne disease burden.
Mosquitoes Genetically Modified to Stop Disease Pass Early Test
22448L. Rapaport, WebMD, 2022-05-09 08:57:53.
Genetically modified mosquitoes released in the U.S. appear to have passed an early test that suggests they might one day help reduce the population of insects that transmit infectious diseases. As part of the test, scientists released nearly 5 million genetically engineered male Aedes aegypti mosquitoes over the course of 7 months in the Florida Keys. Male mosquitoes don’t bite people, and these were also modified so they would transmit a gene to female offspring that causes them to die before they can reproduce. In theory, this means the population of Aedes aegypti mosquitoes would die off over time, so they wouldn’t spread diseases any more. The goal of this pilot project in Florida was to see if these genetically modified male mosquitoes could successfully mate with females in the wild, and to confirm whether their female offspring would indeed die before they could reproduce. On both counts, the experiment was a success, Oxitec, the biotechnology company developing these engineered Aedes aegypti mosquitoes, said in a webina
Transient Introgression of Wolbachia into Aedes aegypti Populations Does Not Elicit an Antibody Response to Wolbachia Surface Protein in Community Members
22655E. Lee, T. Hien Nguyen, T. Yen Nguyen, S. Nam Vu, N. Duong Tran, L. Trung Nghia, Q. Mai Vien, T. Dong Nguyen, R. Kriiger Loterio, I. Iturbe-Ormaetxe, H. A. Flores, S. L. O'Neill, D. Anh Dang, C. P. Simmons and J. E. Fraser, Pathogens, 11. 2022-05-03 06:41:58.
Wolbachia is an endosymbiotic bacterium that can restrict the transmission of human pathogenic viruses by Aedes aegypti mosquitoes. Recent field trials have shown that dengue incidence is significantly reduced when Wolbachia is introgressed into the local Ae. aegypti population. Female Ae. aegypti are anautogenous and feed on human blood to produce viable eggs. Herein, we tested whether people who reside on Tri Nguyen Island (TNI), Vietnam developed antibodies to Wolbachia Surface Protein (WSP) following release of Wolbachia-infected Ae. aegypti, as a measure of exposure to Wolbachia. Paired blood samples were collected from 105 participants before and after mosquito releases and anti-WSP titres were measured by ELISA. We determined no change in anti-WSP titres after ~30 weeks of high levels of Wolbachia-Ae. aegypti on TNI. These data suggest that humans are not exposed to the major Wolbachia surface antigen, WSP, following introgression of Wolbachia-infected Ae. aegypti mosquitoes.
A metapopulation approach to identify targets for Wolbachia-based dengue control
22221A. Reyna-Lara, D. Soriano-Paños, J. H. Arias-Castro, H. J. Martínez and J. Gómez-Gardeñes, Chaos, 32:041105. 2022-04-18 15:42:29.
Over the last decade, the release of Wolbachia-infected Aedes aegypti into the natural habitat of this mosquito species has become the most sustainable and long-lasting technique to prevent and control vector-borne diseases, such as dengue, zika, or chikungunya. However, the limited resources to generate such mosquitoes and their effective distribution in large areas dominated by the Aedes aegypti vector represent a challenge for policymakers. Here, we introduce a mathematical framework for the spread of dengue in which competition between wild and Wolbachia-infected mosquitoes, the cross-contagion patterns between humans and vectors, the heterogeneous distribution of the human population in different areas, and the mobility flows between them are combined. Our framework allows us to identify the most effective areas for the release of Wolbachia-infected mosquitoes to achieve a large decrease in the global dengue prevalence.
Genetically Modified Mosquitoes May Be Released in California, Experts Express Concern
21719Z. Papadakis, NEWSMAX, 2022-04-11 09:25:49.
Millions of genetically engineered mosquitos could soon be set loose in California in an effort to curb the disease-carrying Aedes aegypti mosquito population — but some experts are concerned that it could backfire. On March 7, Oxitec, a private company, obtained a permit from the U.S. Environmental Protection Agency to release its mosquitos in specific districts in Florida and California. The company reasoned that its genetically modified insects could help to save the world half of the world's population from the invasive Aedes aegypti mosquito which, according to Oxitec, increases the risk of transmission of dengue, chikungunya, Zika, yellow fever, and other diseases. "Given the growing health threat this mosquito poses across the U.S., we're working to make this technology available and accessible," Grey Frandsen, CEO of Oxitec, said in a statement. "These pilot programs, wherein we can demonstrate the technology's effectiveness in different climate settings, will play an important role in doing so. We look forward to getting to work this year."
Genetic Stability and Fitness of Aedes aegypti Red-Eye Genetic Sexing Strains With Pakistani Genomic Background for Sterile Insect Technique Applications
22452M. Misbah-ul-Haq, D. O. Carvalho, L. D. de la Fuente, A. A. Augustinos and K. Bourtzis, Frontiers in Bioengineering and Biotechnology, 10. 2022-03-31 09:37:56.
The mosquito species Aedes aegypti is the primary transmitter of viruses that cause endemic diseases like dengue in Pakistan. It is also a cause of other vector-borne diseases like yellow fever, Zika fever, and chikungunya, which significantly impact human health worldwide. In the absence of efficient vaccines (except for yellow fever) or drugs, vector control methods, such as the sterile insect technique (SIT), have been proposed as additional tools for the management of these diseases. Mosquito SIT programs are based on the release of sterile males and it is important female releases to be ideally zero or to be kept at a minimum, since females are the ones that bite, blood-feed and transmit pathogens. Recently, an Ae. aegypti genetic sexing strain (GSS), with and without a recombination-suppressing inversion (Inv35), was developed using the eye color as a selectable marker, with males having black eyes and females red eyes. In the present study, we introgressed the sexing features and the Inv35 of the Ae. aegypti red-eye GSS into the Pakistani genomic background aiming to their future use for SIT applications in the country. Both introgressed strains, the Red-eye GSS-PAK and the Red-eye GSS/Inv35-PAK, were evaluated in respect to their genetic stability and biological quality by assessing parameters like recombination rate, fecundity, fertility, pupal and adult recovery, time of development, pupal weight, survival, and flight ability in comparison with a wild Pakistani population (PAK). The results suggest that the sexing features and the recombination suppression properties of Inv35 were not affected after their introgression into the local genomic background; however, some biological traits of the two newly constructed strains were affected, positively or negatively, suggesting that a thorough quality control analysis should be performed after the introgression of a GSS into a new genomic background prior to its use in SIT field trials or applications. The importance of using GSS with local genomic background for SIT applications against Aedes aegypti is also discussed.
Differential viral RNA methylation contributes to pathogen blocking in Wolbachia-colonized arthropods
20977T. Bhattacharya, L. Yan, J. M. Crawford, H. Zaher, I. L. G. Newton and R. W. Hardy, PLoS Pathogens, 18:e1010393. 2022-03-16 07:40:04.
Arthropod endosymbiont Wolbachia pipientis is part of a global biocontrol strategy to reduce the replication of mosquito-borne RNA viruses such as alphaviruses. We previously demonstrated the importance of a host cytosine methyltransferase, DNMT2, in Drosophila and viral RNA as a cellular target during pathogen-blocking. Here we report a role for DNMT2 in Wolbachia-induced alphavirus inhibition in Aedes species. Expression of DNMT2 in mosquito tissues, including the salivary glands, is elevated upon virus infection. Notably, this is suppressed in Wolbachia-colonized animals, coincident with reduced virus replication and decreased infectivity of progeny virus. Ectopic expression of DNMT2 in cultured Aedes cells is proviral, increasing progeny virus infectivity, and this effect of DNMT2 on virus replication and infectivity is dependent on its methyltransferase activity. Finally, examining the effects of Wolbachia on modifications of viral RNA by LC-MS show a decrease in the amount of 5-methylcytosine modification consistent with the down-regulation of DNMT2 in Wolbachia colonized mosquito cells and animals. Collectively, our findings support the conclusion that disruption of 5-methylcytosine modification of viral RNA is a vital mechanism operative in pathogen blocking. These data also emphasize the essential role of epitranscriptomic modifications in regulating fundamental alphavirus replication and transmission processes.
Wolbachia Impacts Anaplasma Infection in Ixodes scapularis Tick Cells
20436K. M. Skinner, J. Underwood, A. Ghosh, A. S. Oliva Chavez and C. L. Brelsfoard, International Journal of Environmental Research and Public Health, 19. 2022-02-16 08:58:32.
The specific interactions of members of tick bacterial microbiota and their effects on pathogen transmission remains relatively unexplored. Here, we introduced a novel Wolbachia infection type into Ixodes scapularis tick cells and examined the antipathogenic effects on the intracellular pathogen Anaplasma phagocytophilum. An increase in A. phagocytophilum replication was observed in Wolbachia-infected tick cells. However, Wolbachia infection densities decreased when cells were serially passaged and ultimately the infection was lost. Host-cell immune response was also examined as an additional factor that could have affected A. phagocytophilum replication in Wolbachia-infected cells. In early passages post-Wolbachia infection, a decreased immune response was observed, but in later passages of cells with low Wolbachia densities, there was no change in the immune response. The results are discussed in relation to the importance of studying the interactions of the tick microbiota, the host cell, and the pathogen and the development of novel tick and tick-borne disease-control approaches.
Assessing Aedes aegypti candidate genes during viral infection and Wolbachia-mediated pathogen blocking
20315L. T. Sigle, M. Jones, M. Novelo, S. A. Ford, N. Urakova, K. Lymperopoulos, R. T. Sayre, Z. Xi, J. L. Rasgon and E. A. McGraw, Insect Molecular Biology, 2022-02-03 09:03:09.
Abstract One approach to control dengue virus transmission is the symbiont Wolbachia, that limits viral infection in mosquitoes. Despite plans for its widespread use in Aedes aegypti, Wolbachia's mode of action remains poorly understood. Many studies suggest that the mechanism is likely multifaceted, involving aspects of immunity, cellular stress, and nutritional competition. A previous study from our group used artificial selection to identify a new mosquito candidate gene related to viral blocking; alpha-mannosidase-2a (alpha-Mann-2a) with a predicted role in protein glycosylation. Protein glycosylation pathways tend to be involved in complex host-viral interactions; however, the function of alpha-mannosidases has not been described in mosquito-virus interactions. We examined alpha-Mann-2a expression in response to virus and Wolbachia infections and whether reduced gene expression, caused by RNA interference (RNAi), affected viral loads. We show that dengue virus (DENV) infection affects the expression of alpha-Mann-2a in a tissue- and time-dependent manner, whereas Wolbachia infection had no effect. In the midgut, DENV prevalence increased following knockdown of alpha-Mann-2a expression in Wolbachia-free mosquitoes, suggesting that alpha-Mann-2a interferes with infection. Expression knockdown had the same effect on the togavirus chikungunya virus (CHIKV), indicating that alpha-Mann-2a may have broad antivirus effects in the midgut. Interestingly, we were unable to knockdown the expression in Wolbachia-infected mosquitoes. We also provide evidence that alpha-Mann-2a may affect the transcriptional level of another gene predicted to be involved in viral blocking and cell adhesion; cadherin87a. These data support the hypothesis that glycosylation and adhesion pathways may broadly be involved in viral infection in Ae. aegypti. This article is protected by copyright. All rights reserved.
CRISPR Technology Can Eliminate Disease-Spreading Mosquitoes
20133S. Krishana, Now, 2022-01-19 13:10:41.
Scientists have uncovered a new technique they call the “precision-guided sterile insect technique,” or pgSIT. While most CRISPR procedures affect organisms that spread diseases by passing a gene change down generations, this system is more limited. It targets male mosquito genes that are linked to fertility. As a result of changing these genes, any offspring these mosquitoes have would be infertile. “pgSIT is a new scalable genetic control system that uses a CRISPR-based approach to engineer deployable mosquitoes that can suppress populations,” said Omar Akbari, one of the study’s authors. “Males don’t transmit diseases, so the idea is that, as you release more and more sterile males, you can suppress the population without relying on harmful chemicals and insecticides.” But it’s the female population that spreads diseases, so pgSIT targets them, as well. According to the study, the CRISPR technology treatment renders female mosquitoes unable to fly or hold their wings up. It also makes them slower and more lethargic in their movements. Combined, these effects lower the chances that these female mosquitoes will mate or successfully find a blood source and attach to it to spread disease.
Wolbachia: Biological Control Strategy Against Arboviral Diseases
19967I. Mohanty, A. Rath and R. K. Hazra, Genetically Modified and other Innovative Vector Control Technologies, 2021-12-21 11:27:44.
Arboviral diseases like dengue, chikungunya, and Zika are among the major causes of mortality and morbidity in human population. The limited control methods together with lack of antiviral therapies and effective vaccines have paved way for new approaches. One such approach to reduce the ever alarming conflagration of vector-borne diseases is based on biological strategy that reduces or blocks pathogen transmission in the vector. In this context, Wolbachia, an endosymbiont in mosquitoes, is explored as a novel and ecofriendly control strategy. Wolbachia seems to confer resistance to diverse RNA viruses protecting lives from virus-induced mortality. This review envisages the deployment of Wolbachia technology in controlling several arboviral diseases.
Genomic insertion locus and Cas9 expression in the germline affect CRISPR/Cas9-based gene drive performance in the yellow fever mosquito Aedes aegypti
19603W. R. Reid, J. Lin, A. E. Williams, R. Juncu, K. E. Olson and A. W. E. Franz, bioRxiv, 2021.12.08.471839. 2021-12-08 18:56:04.
The yellow fever mosquito Aedes aegypti is a major vector of arthropod-borne viruses, including dengue, chikungunya, and Zika. A novel approach to mitigate arboviral infections is to generate mosquitoes refractory to infection by overexpressing antiviral effector molecules. Such an approach requires a mechanism to spread these antiviral effectors through a population, for example, by using CRISPR/Cas9-based gene drive systems. Here we report an autonomous single-component gene drive system in Ae. aegypti that is designed for persistent population replacement. Critical to the design of a single-locus autonomous gene drive is that the selected genomic locus be amenable to both gene drive and the appropriate expression of the antiviral effector. In our study, we took a reverse engineering approach to target two genomic loci ideal for the expression of antiviral effectors and further investigated the use of three promoters for Cas9 expression (nanos, β2-tubulin, or zpg) for the gene drive. We found that both promoter selection and genomic target site strongly influenced the efficiency of the drive, resulting in 100% inheritance in some crosses. We also observed the formation of inheritable gene drive blocking indels (GDBI) in the genomic locus with the highest levels of gene drive. Overall, our drive system forms a platform for the further testing of driving antipathogen effector genes through Ae. aegypti populations.Competing Interest StatementThe authors have declared no competing interest.
Wolbachia reduces virus infection in a natural population of Drosophila
19437R. Cogni, S. D. Ding, A. C. Pimentel, J. P. Day and F. M. Jiggins, Communications Biology, 4:1327. 2021-11-25 20:42:30.
Wolbachia is a maternally transmitted bacterial symbiont that is estimated to infect approximately half of arthropod species. In the laboratory it can increase the resistance of insects to viral infection, but its effect on viruses in nature is unknown. Here we report that in a natural population of Drosophila melanogaster, individuals that are infected with Wolbachia are less likely to be infected by viruses. By characterising the virome by metagenomic sequencing and then testing individual flies for infection, we found the protective effect of Wolbachia was virus-specific, with the prevalence of infection being up to 15% greater in Wolbachia-free flies. The antiviral effects of Wolbachia may contribute to its extraordinary ecological success, and in nature the symbiont may be an important component of the antiviral defences of insects.
Prevalence and molecular characterization of Wolbachia in field-collected Aedes albopictus, Anopheles sinensis, Armigeres subalbatus, Culex pipiens and Cx. tritaeniorhynchus in China
19078Y. Yang, Y. He, G. Zhu, J. Zhang, Z. Gong, S. Huang, G. Lu, Y. Peng, Y. Meng, X. Hao, C. Wang, J. Sun and S. Shang, PLOS Neglected Tropical Diseases, 15:e0009911. 2021-10-29 20:41:21.
Wolbachia are maternally transmitted intracellular bacteria that can naturally and artificially infect arthropods and nematodes. Recently, they were applied to control the spread of mosquito-borne pathogens by causing cytoplasmic incompatibility (CI) between germ cells of females and males. The ability of Wolbachia to induce CI is based on the prevalence and polymorphism of Wolbachia in natural populations of mosquitoes. In this study, we screened the natural infection level and diversity of Wolbachia in field-collected mosquitoes from 25 provinces of China based on partial sequence of Wolbachia surface protein (wsp) gene and multilocus sequence typing (MLST). Among the samples, 2489 mosquitoes were captured from 24 provinces between July and September, 2014 and the remaining 1025 mosquitoes were collected month-by-month in Yangzhou, Jiangsu province between September 2013 and August 2014. Our results showed that the presence of Wolbachia was observed in mosquitoes of Aedes albopictus (97.1%, 331/341), Armigeres subalbatus (95.8%, 481/502), Culex pipiens (87.0%, 1525/1752), Cx. tritaeniorhynchus (17.1%, 14/82), but not Anopheles sinensis (n = 88). Phylogenetic analysis indicated that high polymorphism of wsp and MLST loci was observed in Ae. albopictus mosquitoes, while no or low polymorphisms were in Ar. subalbatus and Cx. pipiens mosquitoes. A total of 12 unique mutations of deduced amino acid were identified in the wsp sequences obtained in this study, including four mutations in Wolbachia supergroup A and eight mutations in supergroup B. This study revealed the prevalence and polymorphism of Wolbachia in mosquitoes in large-scale regions of China and will provide some useful information when performing Wolbachia-based mosquito biocontrol strategies in China.
A decade of stability for wMel Wolbachia in natural Aedes aegypti populations
19080P. A. Ross, K. L. Robinson, Q. Yang, A. G. Callahan, T. L. Schmidt, J. K. Axford, M. P. Coquilleau, K. M. Staunton, M. Townsend, S. A. Ritchie, M.-J. Lau, X. Gu and A. A. Hoffmann, bioRxiv, 2021.10.27.466190. 2021-10-28 20:41:44.
Mosquitoes carrying Wolbachia endosymbionts are being released in many countries for arbovirus control. The wMel strain of Wolbachia blocks Aedes-borne virus transmission and can spread throughout mosquito populations by inducing cytoplasmic incompatibility. Aedes aegypti mosquitoes carrying wMel were first released into the field in Cairns, Australia, over a decade ago, and with wider releases have resulted in the near elimination of local dengue transmission. The long-term stability of Wolbachia effects is critical for ongoing disease suppression, requiring tracking of phenotypic and genomic changes in Wolbachia infections following releases. We used a combination of field surveys, phenotypic assessments, and Wolbachia genome sequencing to show that wMel has remained stable in its effects for up to a decade in Australian Ae. aegypti populations. Phenotypic comparisons of wMel-infected and uninfected mosquitoes from near-field and long-term laboratory populations suggest limited changes in the effects of wMel on mosquito fitness. Treating mosquitoes with antibiotics used to cure the wMel infection had limited effects on fitness in the next generation, supporting the use of tetracycline for generating uninfected mosquitoes without off-target effects. wMel has a temporally stable within-host density and continues to induce complete cytoplasmic incompatibility. A comparison of wMel genomes from pre-release (2010) and nine years post-release (2020) populations show few genomic differences and little divergence between release locations, consistent with the lack of phenotypic changes. These results indicate that releases of Wolbachia-infected mosquitoes for population replacement are likely to be effective for many years, but ongoing monitoring remains important to track potential evolutionary changes.Competing Interest StatementThe authors have declared no competing interest.
RNA virome diversity and Wolbachia infection in individual Drosophila simulans flies
19073A. S. Ortiz-Baez, M. Shi, A. A. Hoffmann and E. C. Holmes, Journal of General Virology, 102. 2021-10-28 20:29:18.
The endosymbiont bacteria of the genus Wolbachia are associated with multiple mutualistic effects on insect biology, including nutritional and antiviral properties. Members of the genus Wolbachia naturally occur in fly species of the genus Drosophila, providing an operational model host for studying how virome composition may be affected by its presence. Drosophila simulans populations can carry a variety of strains of members of the genus Wolbachia, with the wAu strain associated with strong antiviral protection under experimental conditions. We used D. simulans sampled from the Perth Hills, Western Australia, to investigate the potential virus protective effect of the wAu strain of Wolbachia on individual wild-caught flies. Our data revealed no appreciable variation in virus composition and abundance between individuals infected or uninfected with Wolbachia associated with the presence or absence of wAu. However, it remains unclear whether wAu might affect viral infection and host survival by increasing tolerance rather than inducing complete resistance. These data also provide new insights into the natural virome diversity of D. simulans. Despite the small number of individuals sampled, we identified a repertoire of RNA viruses, including nora virus, galbut virus, thika virus and La Jolla virus, that have been identified in other species of the genus Drosophila. Chaq virus-like sequences associated with galbut virus were also detected. In addition, we identified five novel viruses from the families Reoviridae, Tombusviridae, Mitoviridae and Bunyaviridae. Overall, this study highlights the complex interaction between Wolbachia and RNA virus infections and provides a baseline description of the natural virome of D. simulans.
Novel Symbiotic Genome-Scale Model Reveals Wolbachia’s Arboviral Pathogen Blocking Mechanism in Aedes aegypti
18936N. E. Jiménez, Z. P. Gerdtzen, Á. Olivera-Nappa, J. C. Salgado and C. Conca, mBio, e0156321. 2021-10-13 20:29:16.
Arboviral diseases such as Zika and Dengue have been on the rise mainly due to climate change, and the development of new treatments and strategies to limit their spreading is needed. The use of Wolbachia as an approach for disease control has motivated new research related to the characterization of the mechanisms that underlie its pathogen-blocking properties. In this work, we propose a new approach for studying the metabolic interactions between Aedes aegypti and Wolbachia using genome-scale models, finding that pathogen blocking is mainly influenced by competition for the resources required for Wolbachia and viral replication.
Structural and mechanistic insights into the complexes formed by Wolbachia cytoplasmic incompatibility factors
18908Y. Xiao, H. Chen, H. Wang, M. Zhang, X. Chen, J. M. Berk, L. Zhang, Y. Wei, W. Li, W. Cui, F. Wang, Q. Wang, C. Cui, T. Li, C. Chen, S. Ye, L. Zhang, X. Ji, J. Huang, W. Wang, Z. Wang, M. Hochstrasser and H. Yang, Proceedings of the National Academy of Sciences, 118. 2021-10-09 14:35:56.
Wolbachia bacteria, inherited through the female germ line, infect a large fraction of arthropod species. Many Wolbachia strains manipulate host reproduction, most commonly through cytoplasmic incompatibility (CI). CI, a conditional male sterility, results when Wolbachia-infected male insects mate with uninfected females; viability is restored if the female is similarly infected (called "rescue"). CI is used to help control mosquito-borne viruses such as dengue and Zika, but its mechanisms remain unknown. The coexpressed CI factors CifA and CifB form stable complexes in vitro, but the timing and function of this interaction in the insect are unresolved. CifA expression in the female germ line is sufficient for rescue. We report high-resolution structures of a CI-factor complex, CinA-CinB, which utilizes a unique binding mode between the CinA rescue factor and the CinB nuclease; the structures were validated by biochemical and yeast growth analyses. Importantly, transgenic expression in Drosophila of a nonbinding CinA mutant, designed based on the CinA-CinB structure, suggests CinA expressed in females must bind CinB imported by sperm in order to rescue embryonic viability. Binding between cognate factors is conserved in an enzymatically distinct CI system, CidA-CidB, suggesting universal features in Wolbachia CI induction and rescue.
Wolbachia-Conferred Antiviral Protection Is Determined by Developmental Temperature
18357E. Chrostek, N. Martins, M. S. Marialva and L. Teixeira, mBio, e0292320. 2021-09-08 14:13:12.
Overall, we show that Wolbachia-conferred antiviral protection is temperature dependent, being present or absent depending on the environmental conditions. This interaction likely impacts Wolbachia-host interactions in nature and, as a result, frequencies of host and symbionts in different climates. Dependence of Wolbachia-mediated pathogen blocking on developmental temperature could be used to dissect the mechanistic bases of protection and influence the deployment of Wolbachia to prevent transmission of arboviruses. IMPORTANCE Insects are often infected with beneficial intracellular bacteria. The bacterium Wolbachia is extremely common in insects and can protect them from pathogenic viruses. This effect is being used to prevent transmission of dengue and Zika viruses by Wolbachia-infected mosquitoes. To understand the biology of insects in the wild, we need to discover which factors affect Wolbachia-conferred antiviral protection. Here, we show that the temperature at which insects develop from eggs to adults can determine the presence or absence of antiviral protection. The environment, therefore, strongly influences this insect-bacterium interaction. Our work may help to provide insights into the mechanism of viral blocking by Wolbachia, deepen our understanding of the geographical distribution of host and symbiont, and incentivize further research on the temperature dependence of Wolbachia-conferred protection for control of mosquito-borne disease.
wMel Wolbachia genome remains stable after 7 years in Australian Aedes aegypti field populations
18281K. R. Dainty, J. Hawkey, L. M. Judd, E. C. Pacidônio, J. M. Duyvestyn, D. S. Gonçalves, S. Y. Lin, T. B. O'Donnell, S. L. O'Neill, C. P. Simmons, K. E. Holt and H. A. Flores, Microbial Genomics, 7. 2021-09-02 14:23:53.
Infection of wMel Wolbachia in Aedes aegypti imparts two signature features that enable its application for biocontrol of dengue. First, the susceptibility of mosquitoes to viruses such as dengue and Zika is reduced. Second, a reproductive manipulation is caused that enables wMel introgression into wild-type mosquito populations. The long-term success of this method relies, in part, on evolution of the wMel genome not compromising the critical features that make it an attractive biocontrol tool. This study compared the wMel Wolbachia genome at the time of initial releases and 1-7 years post-release in Cairns, Australia. Our results show the wMel genome remains highly conserved up to 7 years post-release in gene sequence, content, synteny and structure. This work suggests the wMel genome is stable in its new mosquito host and, therefore, provides reassurance on the potential for wMel to deliver long-term public-health impacts.
Wolbachia as translational science: controlling mosquito-borne pathogens
17799E. P. Caragata, H. L. C. Dutra, P. H. F. Sucupira, A. G. A. Ferreira and L. A. Moreira, Trends in Parasitology, 2021-07-22 20:54:55.
In this review we examine how exploiting the Wolbachia?mosquito relationship has become an increasingly popular strategy for controlling arbovirus transmission. Field deployments of Wolbachia-infected mosquitoes have led to significant decreases in dengue virus incidence via high levels of mosquito population suppression and replacement, emphasizing the success of Wolbachia approaches. Here, we examine how improved knowledge of Wolbachia?host interactions has provided key insight into the mechanisms of the essential phenotypes of pathogen blocking and cytoplasmic incompatibility. And we discuss recent studies demonstrating that extrinsic factors, such as ambient temperature, can modulate Wolbachia density and maternal transmission. Finally, we assess the prospects of using Wolbachia to control other vectors and agricultural pest species.
Genetically Modifying Bats Could Prevent the Next Pandemic, Scientists Say
17744G. Dutton, BioSpace, 2021-07-15 13:26:12.
The next COVID pandemic could be prevented by using a gene drive to preemptively edit the genome of bats to prevent them from becoming hosts for coronaviruses, according to a proposal by scientists from Israel’s Interdisciplinary Center (IDC) Herzelia and the National Institutes of Health (NIH). Meanwhile, a team of researchers from Imperial College London is devising a way to prevent gene drives from spreading and conferring heritable, anti-competitive traits to entire populations. The two projects may be in conflict with one another, or the London project may provide a degree of safety that could manage unintended consequences. The IDC/NIH plan, Preventing COVID-59, was published recently on GitHub by Uaniv Erlich of the (IDC) and Daniel Douek of the Vaccine Research Center, National Institute of Allergies and Infectious Diseases at the NIH in the U.S. Its premise is that the SARS-CoV-2 virus – the third such virus to infect humans in the past 20 years – is part of a growing pattern of betacoronaviruses infecting human populations.
Could editing the genomes of bats prevent future coronavirus pandemics? Two scientists think it’s worth a try
17602E. C. Hayden, STAT, 2021-07-01 14:21:06.
Amid the devastating Covid-19 pandemic, two researchers are proposing a drastic way to stop future pandemics: using a technology called a gene drive to rewrite the DNA of bats to prevent them from becoming infected with coronaviruses. The scientists aim to block spillover events, in which viruses jump from infected bats to humans — one suspected source of the coronavirus that causes Covid. Spillover events are thought to have sparked other coronavirus outbreaks as well, including SARS-1 in the early 2000s and Middle East respiratory syndrome (MERS). This appears to be the first time that scientists have proposed using the still-nascent gene drive technology to stop outbreaks by rendering bats immune to coronaviruses, though other teams are investigating its use to stop mosquitoes and mice from spreading malaria and
Preventing COVID-59
17604Y. Erlich and D. Douek, github, 2021-06-30 14:30:52.
SARS-CoV-2 is the third betacoronavirus to enter the human population in the past 20 years, revealing a concerning pattern. Clearly, preventing a future pandemic from such viruses is a critical priority. Previous studies have shown that shRNAs can be powerful suppressors of RNA viruses in transgenic animals and substantially reduce transmission. Thus, we propose the introduction of anti-betacoronavirus shRNAs using CRISPR/CAS9 gene drive into the horseshoe bat population, the natural reservoir of those viruses, to combat this pandemic threat at its source. Importantly, our approach is not expected to create any harm to bats and can benefit other animals in the ecosystem that contract betacoronaviruses from bats. We map the ethical and the technical aspects and suggest guidelines for moving forward with this proposal.
The (Losing) Battle Against Mosquitoes In Texas
17599J. Clayton, Texas Public Radio, 2021-06-26 14:13:20.
Jerry Clayton: Mosquitoes are a fact of life in Texas, and the battle against the pesky biting insects is never ending. But there are some new weapons on the horizon. Zach Adleman is an associate professor of entomology at Texas A&M University. He joins us today. Thanks for being here, Zach.
Targeting conserved sequences circumvents the evolution of resistance in a viral gene drive against human cytomegalovirus
17054M. Walter, R. Perrone and E. Verdin, Journal of virology, 2021-05-19 13:32:53.
Here, we analyze in cell culture experiments the evolution of resistance in a viral gene drive against human cytomegalovirus. We report that, after an initial invasion of the wildtype population, a drive-resistant population is positively selected over time and outcompetes gene drive viruses. However, we show that targeting evolutionary conserved sequences ensures that drive-resistant viruses acquire long-lasting mutations and are durably attenuated. As a consequence, and even though engineered viruses do not stably persist in the viral population, remaining viruses have a replication defect, leading to a long-term reduction of viral levels. This marks an important step toward developing effective gene drives in herpesviruses, especially for therapeutic applications.<b>Importance</b>The use of defective viruses that interfere with the replication of their infectious parent after co-infecting the same cells - a therapeutic strategy known as viral interference - has recently generated a lot of interest. The CRISPR-based system that we recently reported in herpesviruses represents a novel interfering strategy that causes the conversion of wildtype viruses into new recombinant viruses and drives the native viral population to extinction. In this report, we analyzed how targeted viruses evolved resistance against the technology. Through numerical simulations and cell culture experiments with human cytomegalovirus, we show that, after the initial propagation, a resistant viral population is positively selected and outcompetes engineered viruses over time. We show however that targeting evolutionary conserved sequences ensures that resistant viruses are mutated and attenuated, which leads to a long-term reduction of viral levels. This marks an important step toward the development of novel therapeutic strategies against herpesviruses.
Current Effector and Gene-Drive Developments to Engineer Arbovirus-Resistant Aedes aegypti (Diptera: Culicidae) for a Sustainable Population Replacement Strategy in the Field
16596W. R. Reid, K. E. Olson and A. W. E. Franz, J Med Entomol, 2021-03-12 20:35:28.
Conventional mosquito control efforts based on insecticide treatments and/or the use of bednets and window curtains are currently insufficient to reduce arbovirus prevalence in affected regions. Novel, genetic strategies that are being developed involve the genetic manipulation of mosquitoes for population reduction and population replacement purposes. Population replacement aims at replacing arbovirus-susceptible wild-type mosquitoes in a target region with those that carry a laboratory-engineered antiviral effector to interrupt arboviral transmission in the field.
How to engage communities on a large scale? Lessons from World Mosquito Program in Rio de Janeiro, Brazil [version 2; peer review: 1 approved, 2 approved with reservations]
16361G. B. Costa, R. Smithyman, S. L. O'Neill and L. A. Moreira, Gates Open Research, 2021-02-04 19:26:09.
Here we discuss and analyse the framework for community engagement implemented by the WMP in Brazil, during the large-scale deployment of the method in the municipalities of Niterói and Rio de Janeiro, Brazil. Our experience indicates that the community engagement work for arboviruses control should be understood as an opportunity for local development. It is necessary, based on an integrated analysis of the territory, to understand that the actions for arboviruses control could be a catalyst for the necessary socioenvironmental, cultural and public health changes. Furthermore, it is essential to understand that community engagement goes beyond informing or asking for population consent, but it constitutes a possibility for dialogue and exchange between the various stakeholders present in the territories, to build on cooperation for mosquito-borne disease control.
Mosquitoes genetically modified to be resistant to Zika
16259Staff, Lab+Life Scientist, 2021-02-02 15:55:50.
Researchers have wrestled with different strategies for controlling the spread of Zika virus, which is transmitted to humans from female mosquito bites. One approach, which has been approved by the US Environmental Protection Agency, will see more than 750 million genetically modified mosquitoes released into the Florida Keys in 2021 and 2022. These ‘suicide mosquitoes’ are genetically altered to produce offspring that die before emerging into adults and therefore cannot bite humans and spread disease.
Sexual Competitiveness and Induced Egg Sterility by Aedes aegypti and Aedes albopictus Gamma-Irradiated Males: A Laboratory and Field Study in Mexico
17760J. G. Bond, S. Aguirre-Ibáñez, A. R. Osorio, C. F. Marina, Y. Gómez-Simuta, R. Tamayo-Escobar, A. Dor, P. Liedo, D. O. Carvalho and T. Williams, Insects, 12. 2021-01-29 14:44:07.
The sterile insect technique may prove useful for the suppression of mosquito vectors of medical importance in regions where arboviruses pose a serious public health threat. In the present study, we examined the effects of sterilizing irradiation doses across different ratios of fertile:irradiated males on the mating competitiveness of Ae. aegypti and Ae. albopictus under laboratory and field-cage conditions. For both species, the percentage of females inseminated and the number of eggs laid over two gonotrophic cycles varied significantly in mating treatments involving 1:1, 1:5, and 1:10 fertile:irradiated males compared to controls of entirely fertile or entirely irradiated males but was not generally affected by the irradiation dose. Egg hatching was negatively affected in females exposed to increasing proportions of irradiated males in both laboratory and field cages. Male competitiveness (Fried’s index) values varied from 0.19 to 0.58 in the laboratory and were between 0.09 and 1.0 in field cages, depending on th species. Competitiveness values were negatively affected by th eirradiation dose in both species under field-cage conditions, whereas in the laboratory, Ae. albopictus was sensitive to the dose but Ae. aegypti was not. In general, male competitiveness was similar across all mating regimes. Most importantly, induced egg sterility was positively correlated with the proportion of irradiated males present in the mating treatments, reaching a maximum of 88% under field-cage conditions for both Ae. aegypti and Ae. albopictus males treated with 50 and 40 Gy irradiation, respectively. These results indicate that sterile males produced at our facility are suitable and competitive enough for field pilot SIT projects and provide guidance to decide the optimal sterile:fertile ratios.
Genetically-modified mosquitoes key to stopping Zika virus spread
16208University of Missouri, Medical Xpress, 2021-01-26 13:52:44.
Alexander Franz, an associate professor in the MU College of Veterinary Medicine, collaborated with researchers at Colorado State University by using CRISPR gene-editing technology to produce mosquitoes that are unable to replicate Zika virus and therefore cannot infect a human through biting. "We genetically manipulated these mosquitoes by inserting an artificial gene into their genome that triggers one of the immune pathways in the midgut to recognize and destroy the RNA genome of Zika virus," Franz said. "By developing these mosquitoes that are resistant to the virus, the disease cycle is interrupted so transmission to humans can no longer take place." Franz added that the genetic modification is inheritable, so future generations of the altered mosquitoes would be resistant to Zika virus as well.
Targeting evolutionary conserved sequences circumvents the evolution of resistance in a viral gene drive against human cytomegalovirus
15943M. Walter, R. Perrone and E. Verdin, bioRxiv, 2021.01.08.425902. 2021-01-08 16:26:29.
Here, we analyze in cell culture experiments the evolution of resistance in a gene drive against human cytomegalovirus. We report that after an initial invasion of the wildtype population, a drive-resistant population is positively selected over time and outcompetes gene drive viruses. However, we show that targeting evolutionary conserved regions ensures that drive-resistant viruses have a replication defect, leading to a long-term reduction of viral levels. This marks an important step toward developing effective gene drives in viruses, especially for therapeutic applications.
Wolbachia strain wAlbB maintains high density and dengue inhibition following introduction into a field population of Aedes aegypti
15689N. A. Ahmad, M.-V. Mancini, T. H. Ant, J. Martinez, G. M. R. Kamarul, W. A. Nazni, A. A. Hoffmann and S. P. Sinkins, Philosophical Transactions of the Royal Society B: Biological Sciences, 376:20190809. 2020-12-28 15:06:28.
Here, wAlbB-carrying Ae. aegypti collected from the field 20 months after the cessation of releases showed no reduction in Wolbachia density or tissue distribution changes compared to a wAlbB laboratory colony. The wAlbB strain continued to induce complete unidirectional cytoplasmic incompatibility, showed perfect maternal transmission under laboratory conditions, and retained its capacity to inhibit dengue. Additionally, a field-collected wAlbB line was challenged with Malaysian dengue patient blood, and showed significant blocking of virus dissemination to the salivary glands.
CRISPR/Cas9 knockout of female-biased genes AeAct-4 or myo-fem in Ae. aegypti results in a flightless phenotype in female, but not male mosquitoes
15617S. O’Leary and Z. N. Adelman, PLOS Neglected Tropical Diseases, 14:e0008971. 2020-12-18 16:05:35.
Author summary Ae. aegypti is the most important vector of arboviruses throughout the world, and new strategies are urgently needed to add to our existing arsenal of control methods to prevent or halt disease transmission. Importantly, only female Ae. aegypti mosquitoes transmit arboviruses due to their need for vertebrate blood to support egg production. Here, we identify several genes that are critical only for female mosquito flight; genetic ablation of each gene resulted in flightless females, but flying males. Importantly, the female flightless phenotype was completely penetrant (100%), and the use of sex-biased flight muscle proteins appeared to be conserved throughout mosquito evolution. These data could be used to support the development of novel genetic control approaches to reduce the number of biting females across a wide range of disease vector species.
The Antiviral Small-Interfering RNA Pathway Induces Zika Virus Resistance in Transgenic Aedes aegypti
15482A. E. Williams, I. Sanchez-Vargas, W. R. Reid, J. Y. Lin, A. W. E. Franz and K. E. Olson, Viruses, 12:18. 2020-12-15 14:45:29.
We used CRISPR/Cas9 to re-target a previously characterized locus (Chr2:321382225) and engineered mosquitoes expressing an inverted repeat (IR) dsRNA against the NS3/4A region of the ZIKV genome. Small RNA analysis revealed that the IR effector triggered the mosquito's siRNA antiviral pathway in bloodfed females. Nearly complete (90%) inhibition of ZIKV replication was found in vivo in both midguts and carcasses at 7 or 14 days post-infection (dpi). Furthermore, significantly fewer transgenic mosquitoes contained ZIKV in their salivary glands (p = 0.001), which led to a reduction in the number of ZIKV-containing saliva samples as measured by transmission assay. Our work shows that Ae. aegypti innate immunity can be co-opted to engineer mosquitoes resistant to ZIKV.
Reply to: “Enhancement of Aedes aegypti susceptibility to dengue by Wolbachia is not supported”
15264C. Souto-Maior, J. G. King, L. M. Sartori, R. Maciel-de-Freitas and M. G. M. Gomes, Nature Communications, 11:6113. 2020-11-30 19:09:46.
Ant et al.4 claim that concerns with the data and broader analysis make our conclusions misleading. We herein respond to their comments by demonstrating the robustness of our results to different treatments of the data, and expand our arguments for replacing currently adopted methods by those introduced in our paper.
Enhancement of Aedes aegypti susceptibility to dengue by Wolbachia is not supported
15258T. H. Ant, M.-V. Mancini, J. Martinez and S. P. Sinkins, Nature Communications, 11:6111. 2020-11-30 18:48:33.
King et al.3 used DENV infection and transmission modelling to reinterpret experimental data from two previous studies4,5. The authors claimed that wMel Wolbachia increase the mean susceptibility of Ae. aegypti to DENV, contradicting various other studies6,7,8,9,10,11,12. Here, we raise concerns with the experimental approaches used to generate one of the primary datasets on which the modelling is based, and we discuss how these limitations could make some of the original conclusions misleading.
Stable Introduction of Plant-Virus-Inhibiting Wolbachia into Planthoppers for Rice Protection
19719J. T. Gong, Y. Li, T. P. Li, Y. Liang, L. Hu, D. Zhang, C. Y. Zhou, C. Yang, X. Zhang, S. S. Zha, X. Z. Duan, L. A. Baton, X. Y. Hong, A. A. Hoffmann and Z. Xi, Current Biology, 30:4837-4845.e5. 2020-10-10 12:20:07.
Progress has been made in developing the maternally inherited endosymbiotic bacterium Wolbachia as a tool for protecting humans from mosquito-borne diseases. In contrast, Wolbachia-based approaches have not yet been developed for the protection of plants from insect pests and their associated diseases, with a major challenge being the establishment of artificial Wolbachia infections expressing desired characteristics in the hemipterans that transmit the majority of plant viruses. Here, we report stable introduction of Wolbachia into the brown planthopper, Nilaparvata lugens, the most destructive rice pest that annually destroys millions of hectares of staple crops. The Wolbachia strain wStri from the small brown planthopper, Laodelphax striatellus, was transferred to this new host, where it showed high levels of cytoplasmic incompatibility, enabling rapid invasion of laboratory populations. Furthermore, wStri inhibited infection and transmission of Rice ragged stunt virus and mitigated virus-induced symptoms in rice plants, opening up the development of Wolbachia-based strategies against major agricultural pests and their transmitted pathogens
Gene Drives Could Kill Mosquitoes And Suppress Herpesvirus Infections
14588A. Berezow, American Council on Science and Health, 2020-09-30 20:03:47.
A team of researchers writing in the journal Nature Communications has shown that a gene drive can be used to suppress infection with cytomegalovirus, a type of herpesvirus.
Incorporating Characteristics of Gene Drive Engineered Ae. aegypti as Methods to Reduce Dengue and Zika Virus into the Bayesian Network – Relative Risk Model, Using Ponce, Puerto Rico as a Case Study
16097S. R. Eikenbary, WWU Graduate School Collection, 2020-08-10 19:24:02.
The Bayesian network relative risk model can perform the risk assessment of gene drive engineered Ae. aegypti for vector control and as part of an adaptive management strategy to reduce dengue and Zika transmission. This study illustrates how the BN-RRM can integrate gene drive related information within a risk assessment framework suitable for adaptive management of these novel stressors.
Artificial Selection Finds New Hypotheses for the Mechanism of Wolbachia-Mediated Dengue Blocking in Mosquitoes
13600S. A. Ford, I. Albert, S. L. Allen, S. F. Chenoweth, M. Jones, C. Koh, A. Sebastian, L. T. Sigle and E. A. McGraw, Frontiers in Microbiology, 11:1456. 2020-07-07 13:18:12.
We recently used experimental evolution to reveal that Wolbachia-mediated dengue blocking could be selected upon in the A. aegypti host and showed evidence that strong levels of blocking could be maintained by natural selection. In this study, we investigate the genetic variation associated with blocking and use these analyses to generate testable hypotheses surrounding the mechanism of Wolbachia-mediated dengue blocking.
Genetically Modified Mosquitoes Approved For Insect Population Control In The U.S.
12705J. Blum, HUFFPOST, 2020-06-18 20:45:30.
Genetically modified mosquitoes with the ability to prevent other mosquitoes from spreading deadly diseases may be making their way to Florida backyards in the near future. British biotech group Oxitec announced on Tuesday that the company had won both federal and state approval to release its so-called “Friendly” mosquitoes in the U.S. on an experimental trial basis, expected to last until 2022, according to documents provided by the Environmental Protection Agency. The insects will first be released in Monroe County, Florida, and Oxitec has plans to also bring them tT Harris County, Texas.
Florida says ‘this is fine’ to release of genetically modified mosquitoes
12703J. K. Elliot, Global News, 2020-06-18 20:43:23.
That’s the question hanging over a recent decision by state regulators in Florida, which would allow the biotech company Oxitec to unleash hundreds of millions of genetically modified male mosquitoes in the Florida Keys. The lab-altered, patented insects are members of Aedes aegypti, the species of mosquito that spreads diseases such as yellow fever, malaria and chikungunya. However, they’ve been genetically altered to artificially reduce future mosquito populations.
Plan to Release GMO Mosquitoes Moves Ahead
12700A. Dier, newser, 2020-06-18 20:40:30.
A plan to set loose 750 million genetically modified mosquitoes in Florida and Texas will move forward despite concerns from environmentalists who liken it to a "Jurassic Park experiment." The non-biting male Aedes aegypti mosquitoes developed by British biotechnology company Oxitec contain a protein, passed to biting female offspring, that is designed to reduce the insect's chance of surviving into adulthood and therefore prevent the spread of mosquito-borne diseases including Zika and West Nile, reports the Guardian. The EPA has approved two trials, in which the mosquitoes are to be released in Florida's Monroe County this summer and in Texas' Harris County in 2021, though it's facing a lawsuit over its alleged failure to investigate the environmental impact, per the Hill. Facing similar complaints, Florida nonetheless approved its trial on Tuesday.
Genetically engineered mosquitoes get EPA approval for Florida release despite objections from environmental groups
12697S. LaMotte, CNN Health, 2020-06-18 20:37:52.
A genetically modified male mosquito named OX5034 has received both state and federal approval to be released into the Florida Keys now through 2022, against the objection of many local residents and a coalition of environmental advocacy groups. "The administration has used tax dollars and government resources for a Jurassic Park experiment, except without the island," said Jaydee Hanson, policy director at the International Center for Technology Assessment and Center for Food Safety, in a statement.
Florida Keys plans killer insect attack on disease-carrying mosquitoes
12693P. Brinkmann, UPI, 2020-06-18 20:33:39.
The Florida Keys is close to adding a new weapon to help control a mosquito-borne disease -- genetically modified mosquitoes that produce dead offspring. Outbreaks of dengue fever in the Keys in recent years prompted local authorities to consider the genetically modified bugs because the Aedes aegypti mosquito, which spreads that and other diseases, has shown resistance to pesticides.
Plan to release genetically modified mosquitoes in Florida gets go-ahead
12695O. Milman, The Guardian, 2020-06-17 20:35:58.
A plan to release a horde of 750 million genetically modified mosqutioes in Florida and Texas is a step closer to fruition after a state regulator approved the idea, over the objections of many environmentalists.
Fact check: Genetically modified mosquitoes are cleared for release in the US
12443A. Staver, USA Today, 2020-06-13 20:24:44.
The Environmental Protection Agency approved an experimental use permit May 1 that allows Oxitec to release genetically modified mosquitoes in the Florida Keys and Harris County, Texas, where Houston is located.
Editorial Expression of Concern: Transgenic Aedes aegypti Mosquitoes Transfer Genes into a Natural Population
16199B. R. Evans, P. Kotsakiozi, A. L. Costa-Da-Silva, R. S. Ioshino, L. Garziera, M. C. Pedrosa, A. Malavasi, J. F. Virginio, M. L. Capurro and J. R. Powell, Scientific Reports, 10:2. 2020-03-24 15:17:41.
Shortly after publication of this Article in September 2019, the Editors were alerted to concerns regarding the interpretation of the data and some of the conclusions.
Antiviral effectors and gene drive strategies for mosquito population suppression or replacement to mitigate arbovirus transmission by Aedes aegypti
6634A. E. Williams, A. W. E. Franz, W. R. Reid and K. E. Olson, Insects, 11:1-18. 2020-01-12 20:05:33.
The mosquito vector Aedes aegypti transmits arthropod-borne viruses (arboviruses) of medical importance, including Zika, dengue, and yellow fever viruses. Controlling mosquito populations remains the method of choice to prevent disease transmission. Novel mosquito control strategies based on genetically manipulating mosquitoes are being developed as additional tools to combat arbovirus transmission. Genetic control of mosquitoes includes two basic strategies: population suppression and population replacement. The former aims to eliminate mosquito populations while the latter aims to replace wild populations with engineered, pathogen-resistant mosquitoes. In this review, we outline suppression strategies being applied in the field, as well as current antiviral effector genes that have been characterized and expressed in transgenic Ae. aegypti for population replacement. We discuss cutting-edge gene drive technologies that can be used to enhance the inheritance of effector genes, while highlighting the challenges and opportunities associated with gene drives. Finally, we present currently available models that can estimate mosquito release numbers and time to transgene fixation for several gene drive systems. Based on the recent advances in genetic engineering, we anticipate that antiviral transgenic Ae. aegypti exhibiting gene drive will soon emerge; however, close monitoring in simulated field conditions will be required to demonstrate the efficacy and utility of such transgenic mosquitoes.
Transgenic Aedes aegypti Mosquitoes Transfer Genes into a Natural Population
16197B. R. Evans, P. Kotsakiozi, A. L. Costa-da-Silva, R. S. Ioshino, L. Garziera, M. C. Pedrosa, A. Malavasi, J. F. Virginio, M. L. Capurro and J. R. Powell, Scientific Reports, 9:6. 2019-09-10 15:09:04.
We genotyped the release strain and the target Jacobina population before releases began for >21,000 single nucleotide polymorphisms (SNPs). Genetic sampling from the target population six, 12, and 27-30 months after releases commenced provides clear evidence that portions of the transgenic strain genome have been incorporated into the target population. Evidently, rare viable hybrid offspring between the release strain and the Jacobina population are sufficiently robust to be able to reproduce in nature. The release strain was developed using a strain originally from Cuba, then outcrossed to a Mexican population. Thus, Jacobina Ae. aegypti are now a mix of three populations. It is unclear how this may affect disease transmission or affect other efforts to control these dangerous vectors. These results highlight the importance of having in place a genetic monitoring program during such releases to detect un-anticipated outcomes.
Viral gene drive in herpesviruses
5917Walter, M. and E. Verdin, bioRxiv, 2019:717017. 2019-07-30 16:38:14.
Herpesviruses are ubiquitous pathogens in need of novel therapeutic solutions. Current engineered gene drive strategies rely on sexual reproduction, and are thought to be restricted to sexual organisms. Here, we report on the design of a novel gene drive system that allows the spread of an engineered trait in populations of DNA viruses and, in particular, herpesviruses. We describe the successful transmission of a gene drive sequence between distinct strains of human cytomegalovirus (human herpesvirus 5) and show that gene drive viruses can efficiently target and replace wildtype populations in cell culture experiments. Our results indicate that viral gene drives can be used to suppress a viral infection and may represent a novel therapeutic strategy against herpesviruses.
Variability in the durability of CRISPR-Cas immunity
3899Chabas, HN, A.; Meaden, S.; Westra, E. R.; Tremblay, D. M.; Pradier, L.; Lion, S.; Moineau, S.; Gandon, S., Philosophical Transactions of the Royal Society B-Biological Sciences, 374:1-9. 2019-01-17 00:00:00.
The durability of host resistance is challenged by the ability of pathogens to escape the defence of their hosts. Understanding the variability in the durability of host resistance is of paramount importance for designing more effective control strategies against infectious diseases. Here, we study the durability of various clustered regularly interspaced short palindromic repeats-Cas (CRISPR-Cas) alleles of the bacteria Streptococcus thermophilus against lytic phages. We found substantial variability in durability among different resistant bacteria. Since the escape of the phage is driven by a mutation in the phage sequence targeted by CRISPR-Cas, we explored the fitness costs associated with these escape mutations. We found that, on average, escape mutations decrease the fitness of the phage. Yet, the magnitude of this fitness cost does not predict the durability of CRISPR-Cas immunity. We contend that this variability in the durability of resistance may be because of variations in phage mutation rate or in the proportion of lethal mutations across the phage genome. These results have important implications on the coevolutionary dynamics between bacteria and phages and for the optimal deployment of resistance strategies against pathogens and pests. Understanding the durability of CRISPR-Cas immunity may also help develop more effective gene-drive strategies based on CRISPR-Cas9 technology. This article is part of a discussion meeting issue 'The ecology and evolution of prokaryotic CRISPR-Cas adaptive immune systems'.
Engineered resistance to Zika virus in transgenic Aedes aegypti expressing a polycistronic cluster of synthetic small RNAs
3895Buchman, AG, S.; Li, M.; Antoshechkin, I.; Li, H. H.; Wang, H. W.; Chen, C. H.; Klein, M. J.; Duchemin, J. B.; Paradkar, P. N.; Akbari, O. S., Proceedings of the National Academy of Sciences of the United States of America, 116:3656-3661. 2019-01-13 00:00:00.
Recent Zika virus (ZIKV) outbreaks have highlighted the necessity for development of novel vector control strategies to combat arboviral transmission, including genetic versions of the sterile insect technique, artificial infection with Wolbachia to reduce population size and/or vectoring competency, and gene drive-based methods. Here, we describe the development of mosquitoes synthetically engineered to impede vector competence to ZIKV. We demonstrate that a polycistronic cluster of engineered synthetic small RNAs targeting ZIKV is expressed and fully processed in Aedes aegypti, ensuring the formation of mature synthetic small RNAs in the midgut where ZIKV resides in the early stages of infection. Critically, we demonstrate that engineered Ae. aegypti mosquitoes harboring the anti-ZIKV transgene have significantly reduced viral infection, dissemination, and transmission rates of ZIKV. Taken together, these compelling results provide a promising path forward for development of effective genetic-based ZIKV control strategies, which could potentially be extended to curtail other arboviruses.
A secret weapon against Zika and other mosquito-borne diseases
6372Nina Federoff, TEDxMidAtlantic, 2017-07-15 15:37:47.
Where did Zika come from, and what can we do about it? Molecular biologist Nina Fedoroff takes us around the world to understand Zika's origins and how it spread, proposing a controversial way to stop the virus -- and other deadly diseases -- by preventing infected mosquitoes from multiplying.
Genetically Modified Mosquitoes Probably Headed to Florida Keys to Fight Zika
23519T. Elfrink, Miami New Times, 2016-04-11 05:12:57.
The bad news: Zika is coming to Florida. This past Friday, two new cases of the virus linked to serious birth defects were reported in the state, and scientists believe it could spread rapidly come summer as mosquito populations explode. And we're still a long way away from a vaccine for the virus. The good news: Scientists still might have a way to stop its spread — by releasing a swarm of genetically modified mosquitoes across the Florida Keys. Yes, it sounds like the premise of pulpy Michael Crichton novel that ends with a zombie horde infected with GMO mosquito viruses. But it's looking ever more likely to happen. The U.S. Food and Drug Administration (FDA) has given tentative approval to the plan, and the New York Times published a lengthy op-ed this weekend arguing in favor of the plan. And several new polls show that most of the public supports giving it a sho
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