Keywords: Other Symbionts
Evaluating paratransgenesis using engineered symbiotic bacteria for Plasmodium inhibition in mosquito vectors: A systematic review
35538Cleanclay WD, Kernyuy FB, Kintung IF, et al, PLoS Neglected Tropical Diseases, 2026-02-12 16:54:19.
Malaria is one of the key world health problems, especially in sub-Saharan Africa, where the rising resistance to insecticides and antimalarial drugs is posing a threat to the current control mechanisms. New strategies are urgently needed to break the cycle of the spread of Plasmodium parasites by the Anopheles mosquito. A potential solution is paratransgenesis, which involves the genetic modification of naturally occurring mosquito-associated microbes to produce molecules that prevent the development of parasites in the gut of the mosquito. In this systematic review, we examined experimental studies that investigated paratransgenesis as a malaria control measure. Using the PRISMA guideline, we identified ten eligible studies that had engineered bacterial or fungal symbionts to express antiplasmodial effector proteins. These studies showed that feeding mosquitoes a sugar meal containing engineered microbes resulted in a stable colonization of the mosquito midgut. Several of the molecules tested were highly effective in reducing parasite development, with scorpine consistently showing strong transmission-blocking activity, achieving parasite inhibition rates greater than 90% in different experimental systems. Inhibitory effects were also further augmented by the combination of several effector molecules. Overall, the finding suggests that paratransgenesis has strong potential as an additional malaria control measure. Nevertheless, contests concerning ecological safety, microbial stability, and field implementation will need to be solved before large-scale implementation can be done.
The Genomic Arms Race in Mosquito-Borne Diseases: Integrating Entomopathogenic Fungi, Gene Drive, and Symbiont Technologies for Sustainable Vector Control
35503Rajendran Yamini, Pagalahalli Sankaran Shanmugam, Marimuthu Murugan, et al., J Pure Appl Microbiol., 20:53-65. 2026-01-20 09:32:56.
Mosquito-borne diseases such as malaria, dengue, Zika, chikungunya, and lymphatic filariasis continue to impose enormous health and economic burdens worldwide. The traditional reliance on chemical insecticides has been undermined by the rapid evolution of resistance, ecological concerns, and declining efficacy. Next-generation biocontrol strategies are framed within the concept of a “genomic arms race” between mosquitoes, pathogens, and microbial agents. Entomopathogenic fungi are eco-friendly bioinsecticides with demonstrated efficacy in laboratory, semi-field, and transgenic applications. Symbiont-based approaches, particularly those involving Wolbachia, have been evaluated for their ability to reduce vector competence and spread through populations. Parallel advances in CRISPR-based gene drive technologies have provided transformative tools for population suppression and modification, although their deployment is limited by ethical, ecological, and regulatory concerns. An integrated vector management (IVM) framework combining fungi, gene drives, and symbiont-based tools is proposed as the most promising approach for sustainable mosquito management. This multipronged strategy has the potential to reduce disease transmission, delay resistance development, and minimize ecological disruption, paving the way for resilient, eco-friendly solutions against vector-borne diseases.
Monitoring the Capacity of Microsporidia MB Transgenerational Spread in Anopheles arabiensis Populations
35317Boanyah, G. Y., Koekemoer, L. L., Herren, J. K., & Bukhari, T., Insects, 16. 2026-01-05 11:06:39.
Microsporidia MB is a natural symbiont of Anopheles arabiensis Patton, 1905 that blocks Plasmodium transmission without compromising mosquito fitness. Developing this symbiont for malaria control requires a thorough understanding of its transmission dynamics across generations under environmentally relevant conditions. This study aimed to evaluate the transgenerational spread of Microsporidia MB in semi-field settings. From field-collected infected females (G0), several independent replicates of Microsporidia MB-infected An. arabiensis were established and maintained under semi-field conditions for six generations (F1–F6). All the mosquitoes were screened for Microsporidia MB prevalence and infection intensity using qPCR after death. Temperature and humidity and wing size were recorded. Two replicates sustained through F6. Microsporidia MB prevalence and intensity increased from F1 to F3–F4 before declining. A weak positive correlation was observed between prevalence and temperature, but not humidity. Female mosquitoes exhibited higher infection intensities. Female wing size varied across generations, while male size remained consistent. This first study on Microsporidia MB transgenerational transmission under semi-field conditions revealed a pattern of initial increase followed by decline in prevalence and intensity. The stability of male size supports the potential use of infected males in release programs. These findings underscore additional investigations into environmental drivers for mosquito symbiont prevalence.
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.
Emerging challenges for mosquito-borne disease control and the promise of symbiont-based transmission-blocking strategies
35064Gao H, Hu W, Cui C, Wang Y, Zheng Y, Jacobs-Lorena M, et al., PLoS Pathog, 21. 2025-08-26 17:16:49.
Mosquitoes serve as vectors for a variety of pathogens that cause life-threatening diseases, such as malaria, dengue, Zika, and yellow fever. With the rise of antimalarial drug resistance and a lack of therapeutics or prophylactics for dengue and Zika, current disease control strategies rely heavily on mosquito population management. However, the effectiveness of conventional approaches is increasingly compromised, highlighting an urgent need for innovative tools to combat mosquito-borne diseases. One promising strategy for blocking the transmission of these diseases is to populate mosquitoes with anti-pathogen gut symbionts. Here, we discuss the major challenges facing current mosquito-borne disease control efforts and explore how mosquito gut microbiota-based control strategies may address them. We highlight recent advances that may accelerate field applications and offer perspectives on future directions and the translational potential of symbiont-based strategies for mitigating mosquito-borne disease transmission.
An eco-epidemiological model for malaria with Microsporidia MB as bio-control agent
34793Mfangnia CNT, Tonnang HEZ, Tsanou B, Keith Herren J, Modeling Earth Systems and Environment, 11. 2025-04-23 10:24:11.
Microsporidia MB is an endosymbiont which naturally infects Anopheles mosquitoes. Due to its ability to block Plasmodium transmission, it shows potential as a bio-based agent for the control of malaria. Its self-sustainability is promising, as it can spread through both vertical and horizontal transmissions. However, its low prevalence in mosquito populations remains a challenge. We develop an eco-epidemiological mathematical model describing the co-dynamics of Microsporidia MB (within mosquito population) and malaria (within human population). The model is used to assess the potential of Microsporidia MB-infected mosquitoes on the control of malaria infection. The results on the basic reproduction numbers, the stability of the equilibria, and the existence of bifurcations are obtained, providing conditions for the extinction and persistence of MB-infected mosquitoes. We highlight relevant threshold parameters for the elimination and persistence of MB-infected mosquitoes and malaria-infected individuals. Using real data from Kenya, we found that, given a horizontal transmission rate between 0 and 0.5, a minimum vertical rate of 0.55 is required to avoid extinction of MB-infected mosquitoes. The predicted prevalence of MB-infected mosquitoes using transmission rates reported from lab experiments align with the observed low prevalence of MB-infected mosquitoes in the field, thereby validating our model and results. Finally, predictions indicate that increasing MB mosquito infection could effectively control malaria, with target prevalence varying by region: 15% in Highland, 40% on the coast, and 70% in the Lake region. This study offers insights into the use of bio-based vector population replacement solutions to reduce malaria incidence in regions where Microsporidia MB is prevalent.
Wolbachia symbionts control sex in a parasitoid wasp using a horizontally acquired gene
29885Li C, Li CQ, Chen ZB, Liu BQ, Sun X, Wei KH, Li CY, Luan JB., Current Biology, 2024-05-07 15:55:59.
Host reproduction can be manipulated by bacterial symbionts in various ways. Parthenogenesis induction is the most effective type of reproduction manipulation by symbionts for their transmission. Insect sex is determined by regulation of doublesex (dsx) splicing through transformer2 (tra2) and transformer (tra) interaction. Although parthenogenesis induction by symbionts has been studied since the 1970s, its underlying molecular mechanism is unknown. Here we identify a Wolbachia parthenogenesis-induction feminization factor gene (piff) that targets sex-determining genes and causes female-producing parthenogenesis in the haplodiploid parasitoid Encarsia formosa. We found that Wolbachia elimination repressed expression of female-specific dsx and enhanced expression of male-specific dsx, which led to the production of wasp haploid male offspring. Furthermore, we found that E. formosa tra is truncated and non-functional, and Wolbachia has a functional tra homolog, termed piff, with an insect origin. Wolbachia PIFF can colocalize and interact with wasp TRA2. Moreover, Wolbachia piff has coordinated expression with tra2 and dsx of E. formosa. Our results demonstrate the bacterial symbiont Wolbachia has acquired an insect gene to manipulate the host sex determination cascade and induce parthenogenesis in wasps. This study reveals insect-to-bacteria horizontal gene transfer drives the evolution of animal sex determination systems, elucidating a striking mechanism of insect-microbe symbiosis.
Comparative analysis of Wolbachia maternal transmission and localization in host ovaries
29029Michael T.J. Hague, Timothy B. Wheeler, Brandon S. Cooper, bioRxiv, 2024-03-19 13:26:24.
Wolbachia Transinfection Via Embryonic Microinjection
28593Zhang, M., Xi, Z., Methods in Molecular Biology, 2739. 2023-11-29 16:22:25.
The process of transferring Wolbachia from one species to another to establish a stable, maternally inherited infection in the target species is known as transinfection. The success of transinfection is primarily achieved through embryonic microinjection, which is the most direct and efficient means of delivering Wolbachia into the germline of the target species and establishing stable maternal transmission. For the fundamental studies, transinfection is often used to characterize Wolbachia-host interactions, including Wolbachia host range, the role of host or bacterial factors in symbiosis, and evolution of Wolbachia-host associations. For the applied studies, use of transinfection to generate a novel infection in the target species is the first step to build the weapon for both population replacement and population suppression for controlling insect pests or their transmitted diseases. For the primary dengue vector Aedes aegypti and Anopheles vectors of malaria, which either do not naturally carry Wolbachia or are infected with strains that lack necessary features for implementation, transinfection can be established by introducing a novel strain capable of inducing both cytoplasmic incompatibility (CI) and pathogen blocking. For A. albopictus and Culex mosquito species, which naturally harbor CI-inducing Wolbachia, transinfection can be achieved by either introducing a novel strain to generate superinfection or replacing the native infection with a different Wolbachia strain in a symbiont-free line, which is derived from antibiotic treatment of the wild type. Here, we use A. aegypti as an example to describe the Wolbachia transinfection method, which can be adapted to other insect species, such as planthoppers, according to their specific developmental requirements.
Biotechnological Potential of Microorganisms for Mosquito Population Control and Reduction in Vector Competence
28272R. D. Katak, A. M. Cintra, B. C. Burini, O. Marinotti, J. A. Souza-Neto and E. M. Rocha, Insects, 14. 2023-10-30 07:59:35.
Mosquitoes transmit pathogens that cause human diseases such as malaria, dengue fever, chikungunya, yellow fever, Zika fever, and filariasis. Biotechnological approaches using microorganisms have a significant potential to control mosquito populations and reduce their vector competence, making them alternatives to synthetic insecticides. Ongoing research has identified many microorganisms that can be used effectively to control mosquito populations and disease transmission. However, the successful implementation of these newly proposed approaches requires a thorough understanding of the multipronged microorganism-mosquito-;pathogen-environment interactions. Although much has been achieved in discovering new entomopathogenic microorganisms, antipathogen compounds, and their mechanisms of action, only a few have been turned into viable products for mosquito control. There is a discrepancy between the number of microorganisms with the potential for the development of new insecticides and/or antipathogen products and the actual available products, highlighting the need for investments in the intersection of basic research and biotechnology.
A recombinant Aspergillus oryzae fungus transmitted from larvae to adults of Anopheles stephensi mosquitoes inhibits malaria parasite oocyst development
27665L. Kianifard, A. M. Rafiqi, O. Akcakir, A. S. I. Aly, P. F. Billingsley and S. Uysal, Scientific Reports, 13:12177. 2023-08-25 06:27:41.
The control of malaria parasite transmission from mosquitoes to humans is hampered by decreasing efficacies of insecticides, development of drug resistance against the last-resort antimalarials, and the absence of effective vaccines. Herein, the anti-plasmodial transmission blocking activity of a recombinant Aspergillus oryzae (A. oryzae-R) fungus strain, which is used in human food industry, was investigated in laboratory-reared Anopheles stephensi mosquitoes. The recombinant fungus strain was genetically modified to secrete two anti-plasmodial effector peptides, MP2 (midgut peptide 2) and EPIP (enolase-plasminogen interaction peptide) peptides. The transstadial transmission of the fungus from larvae to adult mosquitoes was confirmed following inoculation of A. oryzae-R in the water trays used for larval rearing. Secretion of the anti-plasmodial effector peptides inside the mosquito midguts inhibited oocyst formation of P. berghei parasites. These results indicate that A. oryzae can be used as a paratransgenesis model carrying effector proteins to inhibit malaria parasite development in An. stephensi. Further studies are needed to determine if this recombinant fungus can be adapted under natural conditions, with a minimal or no impact on the environment, to target mosquito-borne infectious disease agents inside their vectors.
Wolbachia has subtle effects on thermal preference in highly inbred Drosophila melanogaster which vary with life stage and environmental conditions
27676A. Strunov, C. Schoenherr and M. Kapun, Scientific Reports, 13:13792. 2023-08-23 07:09:52.
Temperature fluctuations are challenging for ectotherms which are not able to regulate body temperature by physiological means and thus have to adjust their thermal environment via behavior. However, little is yet known about whether microbial symbionts influence thermal preference (Tp) in ectotherms by modulating their physiology. Several recent studies have demonstrated substantial effects of Wolbachia infections on host Tp in different Drosophila species. These data indicate that the direction and strength of thermal preference variation is strongly dependent on host and symbiont genotypes and highly variable among studies. By employing highly controlled experiments, we investigated the impact of several environmental factors including humidity, food quality, light exposure, and experimental setup that may influence Tp measurements in adult Drosophila melanogaster flies. Additionally, we assessed the effects of Wolbachia infection on Tp of Drosophila at different developmental stages, which has not been done before. We find only subtle effects of Wolbachia on host Tp which are strongly affected by experimental variation in adult, but not during juvenile life stages. Our in-depth analyses show that environmental variation has a substantial influence on Tp which demonstrates the necessity of careful experimental design and cautious interpretations of Tp measurements together with a thorough description of the methods and equipment used to conduct behavioral studies.
A mosquito symbiont takes down malaria
27667A. Taglialegna, Nature Reviews Microbiology, 2023-08-14 06:27:59.
Malaria, a parasitic infection transmitted by Anopheles mosquitoes, is globally prevalent. Control strategies for malaria include insecticides and antiparasitic drugs, which target the mosquito vector or the parasite in the human host, respectively. The effectiveness of these methods can be undermined by resistance; hence, new containment approaches are needed. In this study, Huang et al. discover Delftia tsuruhatensis TC1, a mosquito symbiotic bacterium that inhibits parasite development via the secretion of a small molecul
Malaria’s latest foe? Bacteria.
27592M. Coulson, Johns Hopkins, 2023-08-07 06:38:09.
In a new paper published in Science, Marcelo Jacobs-Lorena, PhD, professor emeritus in Molecular Microbiology and Immunology, and his colleagues, in collaboration with researchers at GSK Global Health Medicines R&D, show that a naturally occurring bacterium and a chemical it secretes inhibit the malaria parasite’s development in mosquitoes—meaning they can’t transmit the parasite to humans. The method has distinct advantages: It’s low-tech, easily reproducible, and uses no genetic modification techniques. And, Jacobs-Lorena says, it can work in perfect concert with existing effective control strategies.
A naturally occurring bacteria can stop the malaria parasite right in a mosquito’s gut
27573A. Bhattacharya, Quartz, 2023-08-04 07:55:34.
Scientists at a GlaxoSmithKline (GSK) research facility in Spain discovered that a strain of Delftia tsuruhatensis bacterium, named Tres Cantos 1 (TC1), inhibits the malaria parasite in mosquitoes, known as Plasmodium. Researchers suspected something was going on when the mosquitoes they were using to study malaria were resisting Plasmodium infections. As noted in the study published in the peer-reviewed journal Science yesterday (Aug. 3), TC1 secretes a molecule called harmane that attacks the Plasmodium parasite, which is transmitted to humans by the bite of a mosquito.When the researchers fed the existing strain to other malaria-spreading Anopholes mosquitoes—without any human tinkering like with the genetically-modified microbes—they found the bacteria “drastically reduces malaria parasite burden in the mosquito, potentially reducing transmission to humans significantly,” GSK said in an Aug. 3 press release.
Delftia tsuruhatensis TC1 symbiont suppresses malaria transmission by anopheline mosquitoes
27565W. Huang, J. Rodrigues, E. Bilgo, J. R. Tormo, J. D. Challenger, C. De Cozar-Gallardo, I. Pérez-Victoria, F. Reyes, P. Castañeda-Casado, E. J. Gnambani, D. F. d. S. Hien, M. Konkobo, B. Urones, I. Coppens, A. Mendoza-Losana, L. Ballell, A. Diabate, T. S., Science, 381:533-540. 2023-08-04 07:30:48.
Malaria control demands the development of a wide range of complementary strategies. We describe the properties of a naturally occurring, non?genetically modified symbiotic bacterium, Delftia tsuruhatensis TC1, which was isolated from mosquitoes incapable of sustaining the development of Plasmodium falciparum parasites. D. tsuruhatensis TC1 inhibits early stages of Plasmodium development and subsequent transmission by the Anopheles mosquito through secretion of a small-molecule inhibitor. We have identified this inhibitor to be the hydrophobic molecule harmane. We also found that, on mosquito contact, harmane penetrates the cuticle, inhibiting Plasmodium development. D. tsuruhatensis TC1 stably populates the mosquito gut, does not impose a fitness cost on the mosquito, and inhibits Plasmodium development for the mosquito?s life. Contained field studies in Burkina Faso and modeling showed that D. tsuruhatensis TC1 has the potential to complement mosquito-targeted malaria transmission control. Malaria mosquitoes can act as hosts to several microorganisms, including commensal bacterial species. Huang et al. noticed that some laboratory colonies of anopheline mosquitoes were incapable of transmitting malaria parasites. These insects also harbored a few cells of a bacterium called Delftia tsuruhatensis TC1, which produces a toxic alkaloid called harmane. Bacteria-produced harmane inhibited the development of female Plasmodium parasite gametes in the mosquito gut. Harmane was found to be a contact poison that could also cross the mosquito cuticle to kill developing malaria parasites. Contained field trials in Burkina Faso, coupled with modeling studies, showed that the bacterium has the potential to be deployed in mosquito breeding sites as a component of malaria control. ?Caroline Ash A naturally occurring bacterium of anopheline mosquitoes produces an alkaloid that arrests the development of co-occurring Plasmodium oocytes
Microbe stops mosquitoes from harboring malaria parasite
27567C. Offord, Science, 2023-08-03 07:31:08.
Researchers have tried to use microbes to control mosquito-borne diseases before. The virus-fighting bacterium Wolbachia pipientis has shown particular promise against dengue fever in recent clinical trials and is already used in some areas of the world. But most methods for blocking malaria-causing Plasmodium parasites, which are transmitted by different mosquito species from dengue, have relied on genetically modified bacteria. That’s a major obstacle to regulatory and public acceptance, Barillas-Mury notes, given the unknowns of releasing edited organisms into the wild. The latest news, commentary, and research, free to your inbox daily The bacterium in the new study, published today in Science, inhibits the malaria parasite without any genetic tinkering by humans. Janneth Rodrigues, a scientific lead in global health medicines R&D at GlaxoSmithKline, and colleagues stumbled across the microbe at a GSK research center in Spain, after noticing the mosquitoes they were using for malaria research were getting harder to infect with Plasmodium.
Engineered Gut Symbiotic Bacterium-Mediated RNAi for Effective Control of Anopheles Mosquito Larvae
27559J. J. Ding, C. L. Cui, G. D. Wang, G. Wei, L. Bai, Y. F. Li, P. L. Sun, L. Dong, Z. C. Liu, J. Q. Yun, F. Li, K. Li, L. He and S. B. Wang, Microbiology Spectrum, 2023-07-17 06:46:02.
Anopheles mosquitoes are the primary vectors for the transmission of malaria parasites, which poses a devastating burden on global public health and welfare. The recent invasion of Anopheles stephensi in Africa has made malaria eradication more challenging due to its outdoor biting behavior and widespread resistance to insecticides. To address this issue, we developed a new approach for mosquito larvae control using gut microbiota-mediated RNA interference (RNAi). We engineered a mosquito symbiotic gut bacterium, Serratia fonticola, by deleting its RNase III gene to produce double-stranded RNAs (dsRNAs) in the mosquito larval gut. We found that the engineered S. fonticola strains can stably colonize mosquito larval guts and produce dsRNAs dsMet or dsEcR to activate RNAi and effectively suppress the expression of methoprene-tolerant gene Met and ecdysone receptor gene EcR, which encode receptors for juvenile hormone and ecdysone pathways in mosquitoes, respectively. Importantly, the engineered S. fonticola strains markedly inhibit the development of A. stephensi larvae and leads to a high mortality, providing an effective dsRNA delivery system for silencing genes in insects and a novel RNAi-mediated pest control strategy. Collectively, our symbiont-mediated RNAi (smRNAi) approach offers an innovative and sustainable method for controlling mosquito larvae and provides a promising strategy for combating malaria.IMPORTANCE Mosquitoes are vectors for various diseases, imposing a significant threat to public health globally. The recent invasion of A. stephensi in Africa has made malaria eradication more challenging due to its outdoor biting behavior and widespread resistance to insecticides. RNA interference (RNAi) is a promising approach that uses dsRNA to silence specific genes in pests. This study presents the use of a gut symbiotic bacterium, Serratia fonticola, as an efficient delivery system of dsRNA for RNAi-mediated pest control. The knockout of RNase III, a dsRNA-specific endonuclease gene, in S. fonticola using CRISPR-Cas9 led to efficient dsRNA production. Engineered strains of S. fonticola can colonize the mosquito larval gut and effectively suppress the expression of two critical genes, Met and EcR, which inhibit mosquito development and cause high mortality in mosquito larvae. This study highlights the potential of exploring the mosquito microbiota as a source of dsRNA for RNAi-based pest control. Mosquitoes are vectors for various diseases, imposing a significant threat to public health globally. The recent invasion of A. stephensi in Africa has made malaria eradication more challenging due to its outdoor biting behavior and widespread resistance to insecticides.
Molecular detection of reproductive symbionts and parthenogenesis experiments in Tuta absoluta from Argentina: facing potential for sustainable and specific pest control strategies
26184C. Cagnotti, C. Conte, J. Kramar, S. Lanzavecchia and S. López, Entomologia Experimentalis et Applicata, 2023-06-07 12:24:43.
Tomato leafminer, Tuta absoluta (Meyrick) (Lepidoptera: Gelechiidae), is a key pest of tomato crops. Specific and environmentally friendly control strategies against this pest, such as the sterile insect technique (SIT), are under development and sexual reproduction has emerged as a key focal point to ensure its success. In the present study, we analyzed four representative T. absoluta populations from Argentina, focusing on the molecular detection of endosymbionts potentially associated with reproductive abnormalities and on determining whether parthenogenesis can occur under laboratory conditions. Larvae of T. absoluta were collected in commercial tomato plantations from La Plata and Mar del Plata (both in Buenos Aires province), Bella Vista (Corrientes province), and Yuto (Jujuy province). Tuta absoluta adults were evaluated by standard PCR for the presence of reproductive endosymbionts (Wolbachia sp., Spiroplasma sp., Arsenophonus sp., Cardinium sp., and Rickettsia sp.). For parthenogenesis assays, 20–23 virgin females from each T. absoluta geographic population were life-long isolated in Petri dishes with a tomato leaf. Fecundity, fertility, and female longevity were estimated and compared with results from mated females from each population. Our results showed the presence of a unique endosymbiont (Wolbachia sp. supergroup B, sequence type 41). Fecundity of the unmated females was significantly lower than fecundity of mated females in all T. absoluta populations. In addition, unmated females only laid infertile eggs. In conclusion, the absence of parthenogenesis in T. absoluta from representative geographic populations supports the implementation of SIT and mating disruption as effective control strategies against this pest in Argentina. © 2023 Netherlands Entomological Society.
Holobiont perspectives on tripartite interactions among microbiota, mosquitoes, and pathogens
25208R. Zheng, Q. Wang, R. Wu, P. N. Paradkar, A. A. Hoffmann and G. H. Wang, ISME, 2023-05-25 13:32:28.
Mosquito-borne diseases like dengue and malaria cause a significant global health burden. Unfortunately, current insecticides and environmental control strategies aimed at the vectors of these diseases are only moderately effective in decreasing disease burden. Understanding and manipulating the interaction between the mosquito holobiont (i.e., mosquitoes and their resident microbiota) and the pathogens transmitted by these mosquitoes to humans and animals could help in developing new disease control strategies. Different microorganisms found in the mosquito's microbiota affect traits related to mosquito survival, development, and reproduction. Here, we review the physiological effects of essential microbes on their mosquito hosts; the interactions between the mosquito holobiont and mosquito-borne pathogen (MBP) infections, including microbiota-induced host immune activation and Wolbachia-mediated pathogen blocking (PB); and the effects of environmental factors and host regulation on the composition of the microbiota. Finally, we briefly overview future directions in holobiont studies, and how these may lead to new effective control strategies against mosquitoes and their transmitted diseases.
Combined actions of bacteriophage-encoded genes in Wolbachia-induced male lethality
25420H. Arai, H. Anbutsu, Y. Nishikawa, M. Kogawa, K. Ishii, M. Hosokawa, S. R. Lin, M. Ueda, M. Nakai, Y. Kunimi, T. Harumoto, D. Kageyama, H. Takeyama and M. N. Inoue, iScience, 26:106842. 2023-05-10 08:02:36.
Some Wolbachia endosymbionts induce male killing, whereby male offspring of infected females are killed during development; however, the origin and diversity of the underlying mechanisms remain unclear. In this study, we identified a 76 kbp prophage region specific to male-killing Wolbachia hosted by the moth Homona magnanima. The prophage encoded a homolog of the male-killing gene oscar in Ostrinia moths and the wmk gene that induces various toxicities in Drosophila melanogaster. Upon overexpressing these genes in D. melanogaster, wmk-1 and wmk-3 killed all males and most females, whereas Hm-oscar, wmk-2, and wmk-4 had no impact on insect survival. Strikingly, co-expression of tandemly arrayed wmk-3 and wmk-4 killed 90% of males and restored 70% of females, suggesting their conjugated functions for male-specific lethality. While the male-killing gene in the native host remains unknown, our findings highlight the role of bacteriophages in male-killing evolution and differences in male-killing mechanisms among insects.
A rapidly spreading deleterious aphid endosymbiont that uses horizontal as well as vertical transmission
25082X. Gu, P. A. Ross, A. Gill, Q. Yang, E. Ansermin, S. Sharma, S. Soleimannejad, K. Sharma, A. Callahan, C. Brown, P. A. Umina, T. N. Kristensen and A. A. Hoffmann, Proceedings of the National Academy of Sciences, 120:e2217278120. 2023-04-24 06:34:04.
Endosymbiotic bacteria that live inside the cells of insects are typically only transmitted maternally and can spread by increasing host fitness and/or modifying reproduction in sexual hosts. Transinfections of Wolbachia endosymbionts are now being used to introduce useful phenotypes into sexual host populations, but there has been limited progress on applications using other endosymbionts and in asexual populations. Here, we develop a unique pathway to application in aphids by transferring the endosymbiont Rickettsiella viridis to the major crop pest Myzus persicae. Rickettsiella infection greatly reduced aphid fecundity, decreased heat tolerance, and modified aphid body color, from light to dark green. Despite inducing host fitness costs, Rickettsiella spread rapidly through caged aphid populations via plant-mediated horizontal transmission. The phenotypic effects of Rickettsiella were sensitive to temperature, with spread only occurring at 19 °C and not 25 °C. Body color modification was also lost at high temperatures despite Rickettsiella maintaining a high density. Rickettsiella shows the potential to spread through natural M. persicae populations by horizontal transmission and subsequent vertical transmission. Establishment of Rickettsiella in natural populations could reduce crop damage by modifying population age structure, reducing population growth and providing context-dependent effects on host fitness. Our results highlight the importance of plant-mediated horizontal transmission and interactions with temperature as drivers of endosymbiont spread in asexual insect populations.
A male-killing Wolbachia endosymbiont is concealed by another endosymbiont and a nuclear suppressor
24920K. M. Richardson, P. A. Ross, B. S. Cooper, W. R. Conner, T. Schmidt and A. A. Hoffmann, PLoS Biol, 21:e3001879. 2023-03-22 13:30:17.
Bacteria that live inside the cells of insect hosts (endosymbionts) can alter the reproduction of their hosts, including the killing of male offspring (male killing, MK). MK has only been described in a few insects, but this may reflect challenges in detecting MK rather than its rarity. Here, we identify MK Wolbachia at a low frequency (around 4%) in natural populations of Drosophila pseudotakahashii. MK Wolbachia had a stable density and maternal transmission during laboratory culture, but the MK phenotype which manifested mainly at the larval stage was lost rapidly. MK Wolbachia occurred alongside a second Wolbachia strain expressing a different reproductive manipulation, cytoplasmic incompatibility (CI). A genomic analysis highlighted Wolbachia regions diverged between the 2 strains involving 17 genes, and homologs of the wmk and cif genes implicated in MK and CI were identified in the Wolbachia assembly. Doubly infected males induced CI with uninfected females but not females singly infected with CI-causing Wolbachia. A rapidly spreading dominant nuclear suppressor genetic element affecting MK was identified through backcrossing and subsequent analysis with ddRAD SNPs of the D. pseudotakahashii genome. These findings highlight the complexity of nuclear and microbial components affecting MK endosymbiont detection and dynamics in populations and the challenges of making connections between endosymbionts and the host phenotypes affected by them.
First report of natural Wolbachia infections in mosquitoes from Cuba
24933A. Ruiz, G. Gutiérrez-Bugallo, R. Rodríguez-Roche, L. Pérez, R. González-Broche, L. A. Piedra, L. C. Martínez, Z. Menéndez, A. Vega-Rúa and J. A. Bisset, Acta Tropica, 242:106891. 2023-03-11 08:20:20.
Mosquitoes are extensively responsible for the transmission of pathogens. Novel strategies using Wolbachia could transform that scenario, since these bacteria manipulate mosquito reproduction, and can confer a pathogen transmission-blocking phenotype in culicids. Here, we screened the Wolbachia surface protein region by PCR in eight Cuban mosquito species. We confirmed the natural infections by sequencing and assessed the phylogenetic relationships among the Wolbachia strains detected. We identified four Wolbachia hosts: Aedes albopictus, Culex quinquefasciatus, Mansonia titillans, and Aedes mediovittatus (first report worldwide). Knowledge of Wolbachia strains and their natural hosts is essential for future operationalization of this vector control strategy in Cuba.
Wbm0076, a candidate effector protein of the Wolbachia endosymbiont of Brugia malayi, disrupts eukaryotic actin dynamics
24745M. K. Mills, L. G. McCabe, E. M. Rodrigue, K. F. Lechtreck and V. J. Starai, PLoS Pathogens, 19:e1010777. 2023-02-17 18:26:34.
Brugia malayi, a parasitic roundworm of humans, is colonized by the obligate intracellular bacterium, Wolbachia pipientis. The symbiosis between this nematode and bacterium is essential for nematode reproduction and long-term survival in a human host. Therefore, identifying molecular mechanisms required by Wolbachia to persist in and colonize B. malayi tissues will provide new essential information regarding the basic biology of this endosymbiosis. Wolbachia utilize a Type IV secretion system to translocate so-called "effector" proteins into the cytosol of B. malayi cells to promote colonization of the eukaryotic host. However, the characterization of these Wolbachia secreted proteins has remained elusive due to the genetic intractability of both organisms. Strikingly, expression of the candidate Wolbachia Type IV-secreted effector protein, Wbm0076, in the surrogate eukaryotic cell model, Saccharomyces cerevisiae, resulted in the disruption of the yeast actin cytoskeleton and inhibition of endocytosis. Genetic analyses show that Wbm0076 is a member of the family of Wiskott-Aldrich syndrome proteins (WAS [p]), a well-conserved eukaryotic protein family required for the organization of actin skeletal structures. Thus, Wbm0076 likely plays a central role in the active cell-to-cell movement of Wolbachia throughout B. malayi tissues during nematode development. As most Wolbachia isolates sequenced to date encode at least partial orthologs of wBm0076, we find it likely that the ability of Wolbachia to directly manipulate host actin dynamics is an essential requirement of all Wolbachia endosymbioses, independent of host cell species.
Engineered symbiotic bacteria interfering Nosema redox system inhibit microsporidia parasitism in honeybees
24493H. Lang, H. Wang, H. Wang, X. Xie, X. Hu, X. Zhang and H. Zheng, bioRxiv, 2023.01.13.524015. 2023-01-14 16:42:03.
Nosema ceranae is an emergent microsporidia parasite of the European honey bee (Apis mellifera), which causes serious nosemosis implicated in honeybee colony losses worldwide. N. ceranae is an obligate intracellular eukaryotic parasite that mainly invades the midgut of honeybees. Recent studies find that bee gut microbiota is potentially involved in protecting against parasitism. Here, using laboratory-generated bees mono-associated with gut members, we find that Snodgrassella alvi inhibited microsporidia proliferation, potentially via the stimulation of host oxidant-mediated immune response. Accordingly, N. ceranae employs the thioredoxin and glutathione systems to defend against oxidative stress and maintain a balanced redox equilibrium, which is essential for the infection process. We knocked down the gene expression using nanoparticle-mediated RNA interference, which targets the γ-glutamyl-cysteine synthetase and thioredoxin reductase genes of microsporidia. It significantly reduces the spore load, confirming the importance of the antioxidant mechanism for the intracellular invasion of the N. ceranae parasite. Finally, we genetically modified the symbiotic S. alvi to deliver dsRNA corresponding to the genes involved in the redox system of the microsporidia. The engineered S. alvi induces RNA interference and represses parasite gene expression, thereby inhibits the parasitism by up to 99.8%. Specifically, N. ceranae was most suppressed by the recombinant strain corresponding to the glutathione synthetase or by a mixture of bacteria expressing variable dsRNA. Our findings extend our previous understanding of the protection of gut symbionts against N. ceranae and provide a symbiont-mediated RNAi system for inhibiting microsporidia infection in honeybees.Competing Interest StatementThe authors have declared no competing interest.
Cell-based analysis reveals that sex-determining gene signals in Ostrinia are pivotally changed by male-killing Wolbachia
24288B. Herran, T. N. Sugimoto, K. Watanabe, S. Imanishi, T. Tsuchida, T. Matsuo, Y. Ishikawa and D. Kageyama, PNAS Nexus, pgac293. 2022-12-13 08:59:49.
Wolbachia, a maternally transmitted bacterium, shows male-killing, an adaptive phenotype for cytoplasmic elements, in various arthropod species during the early developmental stages. In lepidopteran insects, lethality of males is accounted for by improper dosage compensation in sex-linked genes owing to Wolbachia-induced feminization. Herein, we established Ostrinia scapulalis cell lines that retained sex specificity per the splicing pattern of the sex-determining gene doublesex (Osdsx). We found that Wolbachia transinfection in male cell lines enhanced the female-specific splice variant of Osdsx (OsdsxF) while suppressing the male-specific variant (OsdsxM), indicating that Wolbachia affects sex-determining gene signals even in vitro. Comparative transcriptome analysis isolated only two genes that behave differently upon Wolbachia infection. The two genes were respectively homologous to Masculinizer (BmMasc) and zinc finger-2 (Bmznf-2), male-specifically expressed sex-determining genes of the silkworm Bombyx mori that encode CCCH-type zinc finger motif proteins. By using cultured cells and organismal samples, OsMasc and Osznf-2 were found to be sex-determining genes of O. scapulalis that are subjected to sex-specific alternative splicing depending upon the chromosomal sex, developmental stage, and infection status. Overall, our findings expound the cellular autonomy in insect sex determination and the mechanism through which sex is manipulated by intracellular selfish microbes.
Tolerance-conferring defensive symbionts and the evolution of parasite virulence
24126C. A. Smith and B. Ashby, bioRxiv, 2022-12-05 09:03:54.
Defensive symbionts in the host microbiome can confer protection from infection or reduce the harms of being infected by a parasite. Defensive symbionts are therefore promising agents of biocontrol that could be used to control or ameliorate the impact of infectious diseases. Previous theory has shown how symbionts can evolve along the parasitism-mutualism continuum to confer greater or lesser protection to their hosts, and in turn how hosts may coevolve with their symbionts to potentially form a mutualistic relationship. However, the consequences of introducing a defensive symbiont for parasite evolution and how the symbiont may coevolve with the parasite have yet to be explored theoretically. Here, we investigate the ecological and evolutionary implications of introducing a tolerance-conferring defensive symbiont into an established host-parasite system. We show that while the defensive symbiont may initially have a positive impact on the host population, parasite and symbiont evolution tend to have a net negative effect on the host population in the long-term. This is because the introduction of the defensive symbiont always selects for an increase in parasite virulence and may cause diversification into high- and low-virulence strains. Even if the symbiont experiences selection for greater host protection, this simply increases selection for virulence in the parasite, resulting in a net negative effect on the host population. Our results therefore suggest that tolerance-conferring defensive symbionts may be poor biocontrol agents for population-level infectious disease control.Competing Interest StatementThe authors have declared no competing interest.
A comprehensive overview of the existing microbial symbionts in mosquito vectors: An important tool for impairing pathogentransmission
24088V. Vandana, M. P. Kona, J. Kumar, O. P. Singh and K. C. Pandey, Experimental Parasitology, 243. 2022-11-30 09:06:12.
The emergence of drug-resistant parasites and/or insecticide-resistant mosquito vectors necessitates developing alternative tools that either supplement or replace the conventional malaria control strategies. Trans-infecting the mosquito vector with symbionts that can either compete with a targeted pathogen or manipulate the host biology by reducing its vectorial capacity could be a promising and innovative biological approach for the control of infectious diseases This idea could be utilized to develop a novel and efficient vector control strategy; symbionts are dispersed into vector populations to reduce their ability to transmit human pathogens. Here, we reported the natural existence of Microsporidian (an obligate fungus) in the field-collected An. stephensi mosquito. However, laboratory-reared An. stephensi and An. culicifacies did not exhibit microsporidian infection. Similarly, 16s rRNA PCR identified -1kb amplicons in laboratory-reared An. stephensi and An. culicifacies, indicating the presence of naturally residing different bacterial species. DNA sequencing of these amplicons revealed the identities of different bacteria which are not well-characterized in terms of plasmodia-interaction activity in the Indian malaria vector. This article summarizes an overview of the previously studied microbial symbionts for their role in Plasmodium transmission along with a list of new or unexplored symbionts in the disease transmitting mosquito vectors. The summarized information could be utilized to explore such microbial symbionts for their role in Plasmodium-transmission biology in-depth and implementation in the malaria control interventions globally.
A Wolbachia factor for male killing in lepidopteran insects
24016S. Katsuma, K. Hirota, N. Matsuda-Imai, T. Fukui, T. Muro, K. Nishino, H. Kosako, K. Shoji, H. Takanashi, T. Fujii, S.-i. Arimura and T. Kiuchi, Nature Communications, 13:6764. 2022-11-14 12:59:35.
Bacterial symbionts, such as Wolbachia species, can manipulate the sexual development and reproduction of their insect hosts. For example, Wolbachia infection induces male-specific death in the Asian corn borer Ostrinia furnacalis by targeting the host factor Masculinizer (Masc), an essential protein for masculinization and dosage compensation in lepidopteran insects. Here we identify a Wolbachia protein, designated Oscar, which interacts with Masc via its ankyrin repeats. Embryonic expression of Oscar inhibits Masc-induced masculinization and leads to male killing in two lepidopteran insects, O. furnacalis and the silkworm Bombyx mori. Our study identifies a mechanism by which Wolbachia induce male killing of host progeny.
Wolbachia inhibits ovarian formation and increases blood feeding rate in female Aedes aegypti
23858M.-J. Lau, P. A. Ross, N. M. Endersby-Harshman, Q. Yang and A. A. Hoffmann, PLOS Neglected Tropical Diseases, 16:e0010913. 2022-11-11 09:59:48.
Author summary Wolbachia bacteria reduce the transmission of dengue and other arboviruses transmitted by their mosquito hosts once they have invaded host populations. This invasion process is being undertaken in Aedes aegypti mosquito populations throughout the world but can be slowed by deleterious fitness costs that the Wolbachia bacteria have on their mosquito hosts. Here we investigate in detail a recently identified cost where one of the Wolbachia strains being used in invasions causes infertility in females when the females develop from eggs that have been stored for a period. We show that slowed larval development (through food reduction) can cause the same phenomenon and that the emerged females lack functional ovaries. These infertile females also take blood meals more often. The results have implications for the successful invasion of Wolbachia under ecological circumstances where the mosquito eggs have to persist in the absence of water for a period and where larval food is limiting.
Combining transgenesis with paratransgenesis to fight malaria
23802W. Huang, J. Vega-Rodriguez, C. Kizito, S.-J. Cha and M. Jacobs-Lorena, eLife, 11:e77584. 2022-10-25 06:30:55.
Malaria is among the deadliest infectious diseases, and Plasmodium, the causative agent, needs to complete a complex development cycle in its vector mosquito for transmission to occur. Two promising strategies to curb transmission are transgenesis, consisting of genetically engineering mosquitoes to express antimalarial effector molecules, and paratransgenesis, consisting of introducing into the mosquito commensal bacteria engineered to express antimalarial effector molecules. Although both approaches restrict parasite development in the mosquito, it is not known how their effectiveness compares. Here we provide an in-depth assessment of transgenesis and paratransgenesis and evaluate the combination of the two approaches. Using the Q-system to drive gene expression, we engineered mosquitoes to produce and secrete two effectors – scorpine and the MP2 peptide – into the mosquito gut and salivary glands. We also engineered Serratia, a commensal bacterium capable of spreading through mosquito populations to secrete effectors into the mosquito gut. Whereas both mosquito-based and bacteria-based approaches strongly reduced the oocyst and sporozoite intensity, a substantially stronger reduction of Plasmodium falciparum development was achieved when transgenesis and paratransgenesis were combined. Most importantly, transmission of Plasmodium berghei from infected to naïve mice was maximally inhibited by the combination of the two approaches. Combining these two strategies promises to become a powerful approach to combat malaria.
Hidden endosymbionts: A male-killer concealed by another endosymbiont and a nuclear suppressor
23793K. M. Richardson, P. A. Ross, B. S. Cooper, W. R. Conner, T. Schmidt and A. A. Hoffmann, bioRxiv, 2022.10.19.512817. 2022-10-22 06:41:08.
Maternally transmitted endosymbiotic bacteria that cause male killing (MK) have only been described from a few insects, but this may reflect challenges in their detection rather than a rarity of MK. Here we identify MK Wolbachia in populations of Drosophila pseudotakahashii, present at a low frequency (around 4%) in natural populations and previously undetected due to a different fixed Wolbachia strain in this species expressing a different reproductive manipulation, cytoplasmic incompatibility (CI). The MK phenotype was eliminated after tetracycline treatment that removed Wolbachia. Molecular analyses indicated the MK phenotype to be expressed when a second Wolbachia strain was present alongside the CI Wolbachia. A genomic analysis highlighted Wolbachia regions diverged between the strains involving 17 genes and also identified the Wolbachia as representing an outgroup to a clade of Wolbachia infecting melanogaster-group species, including wRi-like and wMel-like strains. Doubly infected males induced CI with uninfected females but not females singly infected with CI-causing Wolbachia. The MK phenotype manifested at the larval stage and was transmitted maternally at a high fidelity but with occasional loss of the MK Wolbachia strain. A rapidly spreading dominant nuclear suppressor genetic element affecting MK was identified through backcrossing and subsequent analysis with ddRAD SNPs of the D. pseudotakahashii genome. These findings highlight the complexity of nuclear and microbial components affecting MK endosymbiont detection and dynamics in populations, and the challenges of making connections between endosymbionts and the host phenotypes affected by them.Competing Interest StatementThe authors have declared no competing interest.
An evaluation of fusion partner proteins for paratransgenesis in Asaia bogorensis
24274C. Grogan, M. Bennett and D. J. Lampe, Plos One, 17:18. 2022-09-01 14:11:14.
Mosquitoes transmit many pathogens responsible for human diseases, such as malaria which is caused by parasites in the genus Plasmodium. Current strategies to control vector-transmitted diseases are increasingly undermined by mosquito and pathogen resistance, so additional methods of control are required. Paratransgenesis is a method whereby symbiotic bacteria are genetically modified to affect the mosquito's phenotype by engineering them to deliver effector molecules into the midgut to kill parasites. One paratransgenesis candidate is Asaia bogorensis, a Gram-negative bacterium colonizing the midgut, ovaries, and salivary glands of Anopheles sp. mosquitoes. Previously, engineered Asaia strains using native signals to drive the release of the antimicrobial peptide, scorpine, fused to alkaline phosphatase were successful in significantly suppressing the number of oocysts formed after a blood meal containing P. berghei. However, these strains saw high fitness costs associated with the production of the recombinant protein. Here, we report evaluation of five different partner proteins fused to scorpine that were evaluated for effects on the growth and fitness of the transgenic bacteria. Three of the new partner proteins resulted in significant levels of protein released from the Asaia bacterium while also significantly reducing the prevalence of mosquitoes infected with P. berghei. Two partners performed as well as the previously tested Asaia strain that used alkaline phosphatase in the fitness analyses, but neither exceeded it. It may be that there is a maximum level of fitness and parasite inhibition that can be achieved with scorpine being driven constitutively, and that use of a Plasmodium specific effector molecule in place of scorpine would help to mitigate the stress on the symbionts.
Bacterial supergroup-specific “cost” of Wolbachia infections in Nasonia vitripennis
23640A. Tiwary, R. Babu, R. Sen and R. Raychoudhury, Ecology and Evolution, 2022-09-01 08:00:24.
Abstract The maternally inherited endosymbiont, Wolbachia, is known to alter the reproductive biology of its arthropod hosts for its own benefit and can induce both positive and negative fitness effects in many hosts. Here, we describe the effects of the maintenance of two distinct Wolbachia infections, one each from supergroups A and B, on the parasitoid host Nasonia vitripennis. We compare the effect of Wolbachia infections on various traits between the uninfected, single A-infected, single B-infected, and double-infected lines with their cured versions. Contrary to some previous reports, our results suggest that there is a significant cost associated with the maintenance of Wolbachia infections where traits such as family size, fecundity, longevity, and rates of male copulation are compromised in Wolbachia-infected lines. The double Wolbachia infection has the most detrimental impact on the host as compared to single infections. Moreover, there is a supergroup-specific negative impact on these wasps as the supergroup B infection elicits the most pronounced negative effects. These negative effects can be attributed to a higher Wolbachia titer seen in the double and the single supergroup B infection lines when compared to supergroup A. Our findings raise important questions on the mechanism of survival and maintenance of these reproductive parasites in arthropod hosts.
Rational engineering of a synthetic insect-bacterial mutualism
23473Y. Su, H.-C. Lin, L. S. Teh, F. Chevance, I. James, C. Mayfield, K. G. Golic, J. A. Gagnon, O. Rog and C. Dale, Current Biology, 2022-08-29 07:21:11.
Summary Many insects maintain mutualistic associations with bacterial endosymbionts, but little is known about how they originate in nature. In this study, we describe the establishment and manipulation of a synthetic insect-bacterial symbiosis in a weevil host. Following egg injection, the nascent symbiont colonized many tissues, including prototypical somatic and germinal bacteriomes, yielding maternal transmission over many generations. We then engineered the nascent symbiont to overproduce the aromatic amino acids tyrosine and phenylalanine, which facilitate weevil cuticle strengthening and accelerated larval development, replicating the function of mutualistic symbionts that are widely distributed among weevils and other beetles in nature. Our work provides empirical support for the notion that mutualistic symbioses can be initiated in insects by the acquisition of environmental bacteria. It also shows that certain bacterial genera, including the Sodalis spp. used in our study, are predisposed to develop these associations due to their ability to maintain benign infections and undergo vertical transmission in diverse insect hosts, facilitating the partner-fidelity feedback that is critical for the evolution of obligate mutualism. These experimental advances provide a new platform for laboratory studies focusing on the molecular mechanisms and evolutionary processes underlying insect-bacterial symbiosis.
Biologists engineered insect-bacterial mutualism in ‘bucket list’ achievement
23475Annonymous, @THEU, 2022-08-23 07:28:20.
A new paper in Current Biology authored by Crystal Su and other collaborators in the School of Biological Sciences describes the development of a novel, synthetic insect-bacterial symbiosis that is sustained through many insect generations by transovarial bacterial transmission. The symbiotic bacteria express a red fluorescent protein that is visible through the insect cuticle, facilitating characterization of the mechanics of infection and transmission in insect tissues and cells. In addition, Su et. al.engineered the bacteria to modify their ability to synthesize aromatic amino acids, which are used by the insect host to fuel cuticle strengthening. Correspondingly, insects maintaining bacteria that overproduce these nutrients exhibited stronger cuticles, signifying mutualistic function. The establishment of this synthetic symbiosis will facilitate detailed molecular genetic analysis of symbiotic interactions and presents a foundation for the use of genetically modified symbionts in the engineering of insects that transmit diseases of medical and agricultural importance. The paper is titled “Rational engineering of a synthetic insect-bacterial mutualism.”
Aedes aegypti and Ae. albopictus microbiome/virome: new strategies for controlling arboviral transmission?
23413M. Gómez, D. Martinez, M. Muñoz and J. D. Ramírez, Parasites and Vectors, 15:287. 2022-08-09 12:21:12.
Aedes aegypti and Aedes albopictus are the main vectors of highly pathogenic viruses for humans, such as dengue (DENV), chikungunya (CHIKV), and Zika (ZIKV), which cause febrile, hemorrhagic, and neurological diseases and remain a major threat to global public health. The high ecological plasticity, opportunistic feeding patterns, and versatility in the use of urban and natural breeding sites of these vectors have favored their dispersal and adaptation in tropical, subtropical, and even temperate zones. Due to the lack of available treatments and vaccines, mosquito population control is the most effective way to prevent arboviral diseases. Resident microorganisms play a crucial role in host fitness by preventing or enhancing its vectorial ability to transmit viral pathogens. High-throughput sequencing and metagenomic analyses have advanced our understanding of the composition and functionality of the microbiota of Aedes spp. Interestingly, shotgun metagenomics studies have established that mosquito vectors harbor a highly conserved virome composed of insect-specific viruses (ISV). Although ISVs are not infectious to vertebrates, they can alter different phases of the arboviral cycle, interfering with transmission to the human host. Therefore, this review focuses on the description of Ae. aegypti and Ae. albopictus as vectors susceptible to infection by viral pathogens, highlighting the role of the microbiota-virome in vectorial competence and its potential in control strategies for new emerging and re-emerging arboviruses.
Sexual transmission of Anopheles gambiae densovirus (AgDNV) leads to disseminated infection in mated females
23030K. L. Werling, R. M. Johnson, H. C. Metz and J. L. Rasgon, Parasites and Vectors, 15:219. 2022-06-20 07:20:44.
Anopheles gambiae densovirus (AgDNV) is an insect-specific, single-stranded DNA virus that infects An. gambiae sensu stricto (s.s.), the major mosquito species responsible for transmitting malaria parasites throughout sub-Saharan Africa. AgDNV is a benign virus that is very specific to its mosquito host and therefore has the potential to serve as a vector control tool via paratransgenesis (genetic modification of mosquito symbionts) to limit transmission of human pathogens. Prior to being engineered into a control tool, the natural transmission dynamics of AgDNV between An. gambiae mosquitoes needs to be fully understood. Additionally, improved knowledge of AgDNV infection in male mosquitoes is needed. In the study presented here, we examined the tissue tropism of AgDNV in the male reproductive tract and investigated both venereal and vertical transmission dynamics of the virus.
Male-killing-associated bacteriophage WO identified from comparisons of Wolbachia endosymbionts of Homona magnanima
22901H. Arai, H. Anbutsu, Y. Nishikawa, M. Kogawa, K. Ishii, M. Hosokawa, S.-R. Lin, M. Ueda, M. Nakai, Y. Kunimi, T. Harumoto, D. Kageyama, H. Takeyama and M. N. Inoue, bioRxiv, 2022-06-13 06:22:33.
The origin and mechanism of male-killing, an advantageous strategy employed by maternally transmitted symbionts such as Wolbachia, remain unclear. We compared genomes of four Wolbachia strains derived from Homona magnanima, a male-killing strain wHm-t (1.5 Mb), and three non-male-killing strains, wHm-a (1.1 Mb), wHm-b (1.3 Mb), and wHm-c (1.4 Mb). A wHm-t-specific 76-kbp prophage region harboured two tandemly arrayed WO-mediated killing (wmk) gene homologs (wmk-1/wmk-2 and wmk-3/wmk-4). Of these, wmk-1 or wmk-3 killed almost all Drosophila melanogaster individuals when transgenically overexpressed. Dual expression of wmk-3 and wmk-4 killed all males and rescued females. We propose a novel hypothesis wherein horizontally transmitted proto-Wolbachia with a single wmk killed both sexes, and tandem duplication of wmk allowed an evolutionary transition to a vertically transmitted symbiont, causing male-killing. Our study highlights the bacteriophage as a critical driver of the evolution of male-killing and argues for a conserved male-killing mechanism in diverse insects.Competing Interest StatementThe authors have declared no competing interest.
Wolbachia interacts with the microbiome to shape fitness-associated traits during seasonal adaptation in Drosophila melanogaster
22693L. P. Henry, M. Fernandez, S. Wolf and J. Ayroles, bioRxiv, 2022.05.31.494239. 2022-06-01 14:39:02.
The microbiome contributes to many different host traits, but its role in host adaptation remains enigmatic. The fitness benefits of the microbiome often depend on ecological conditions, but fluctuations in both the microbiome and environment modulate these fitness benefits. Moreover, vertically transmitted bacteria might constrain the ability of both the microbiome and host to respond to changing environments. Drosophila melanogaster provides an excellent system to investigate the evolutionary effects of interactions between the microbiome and the environment. To address this question, we created field mesocosms of D. melanogaster undergoing seasonal adaptation with and without the vertically transmitted bacteria, Wolbachia pipientis. Sampling temporal patterns in the microbiome revealed that Wolbachia constrained microbial diversity. Furthermore, interactions between Wolbachia and the microbiome contributed to fitness-associated traits. Wolbachia often exerted negative fitness effects on hosts, and the microbiome modulated these effects. Our work supports recent theoretical advances suggesting that hosts in temporally fluctuating environments benefit from flexible microbial associations with low transmission fidelity--specifically when changes in the microbiome can better enable host phenotypes to match environment change. We conclude by exploring the consequences of complex interactions between Wolbachia and the microbiome for our understanding of eco-evolutionary processes and the utility of Wolbachia in combating vector-borne disease.Competing Interest StatementThe authors have declared no competing interest.
New weapons to fight malaria transmission: A historical view
22487W. Huang, S.-J. Cha and M. Jacobs-Lorena, Entomological Research, 2022-05-02 07:19:32.
The stagnation of our fight against malaria in recent years, mainly due to the development of mosquito insecticide resistance, argues for the urgent development of new weapons. The dramatic evolution of molecular tools in the last few decades led to a better understanding of parasite?mosquito interactions and coalesced in the development of novel tools namely, mosquito transgenesis and paratransgenesis. Here we provide a historical view of the development of these new tools and point to some remaining challenges for their implementation in the field.
Wolbachia endosymbionts in two Anopheles species indicates independent acquisitions and lack of prophage elements
21788S. Quek, L. Cerdeira, C. L. Jeffries, S. Tomlinson, T. Walker, G. L. Hughes and E. Heinz, Microbial Genomics, 8. 2022-04-21 06:52:58.
Wolbachia is a genus of obligate bacterial endosymbionts that infect a diverse range of arthropod species as well as filarial nematodes, with its single described species, Wolbachia pipientis, divided into several ‘supergroups’ based on multilocus sequence typing. Wolbachia strains in mosquitoes have been shown to inhibit the transmission of human pathogens, including Plasmodium malaria parasites and arboviruses. Despite their large host range, Wolbachia strains within the major malaria vectors of the Anopheles gambiae and Anopheles funestus complexes appear at low density, established solely on PCR-based methods. Questions have been raised as to whether this represents a true endosymbiotic relationship. However, recent definitive evidence for two distinct, high-density strains of supergroup B Wolbachia within Anopheles demeilloni and Anopheles moucheti has opened exciting possibilities to explore naturally occurring Wolbachia endosymbionts in Anopheles for biocontrol strategies to block Plasmodium transmission. Here, we utilize genomic analyses to demonstrate that both Wolbachia strains have retained all key metabolic and transport pathways despite their smaller genome size, with this reduction potentially attributable to degenerated prophage regions. Even with this reduction, we confirmed the presence of cytoplasmic incompatibility (CI) factor genes within both strains, with wAnD maintaining intact copies of these genes while the cifB gene was interrupted in wAnM, so functional analysis is required to determine whether wAnM can induce CI. Additionally, phy logenetic analysis indicates that these Wolbachia strains may have been introduced into these two Anopheles species via horizontal transmission events, rather than by ancestral acquisition and subsequent loss events in the Anopheles gambiae species complex. These are the first Wolbachia genomes, to our knowledge, that enable us to study the relationship between natural strain Plasmodium malaria parasites and their anopheline hosts.
Overview of paratransgenesis as a strategy to control pathogen transmission by insect vectors
21579N. A. Ratcliffe, J. P. Furtado Pacheco, P. Dyson, H. C. Castro, M. S. Gonzalez, P. Azambuja and C. B. Mello, Parasites and Vectors, 15:112. 2022-03-31 12:47:27.
This article presents an overview of paratransgenesis as a strategy to control pathogen transmission by insect vectors. It first briefly summarises some of the disease-causing pathogens vectored by insects and emphasises the need for innovative control methods to counter the threat of resistance by both the vector insect to pesticides and the pathogens to therapeutic drugs. Subsequently, the state of art of paratransgenesis is described, which is a particularly ingenious method currently under development in many important vector insects that could provide an additional powerful tool for use in integrated pest control programmes. The requirements and recent advances of the paratransgenesis technique are detailed and an overview is given of the microorganisms selected for genetic modification, the effector molecules to be expressed and the environmental spread of the transgenic bacteria into wild insect populations. The results of experimental models of paratransgenesis developed with triatomines, mosquitoes, sandflies and tsetse flies are analysed. Finally, the regulatory and safety rules to be satisfied for the successful environmental release of the genetically engineered organisms produced in paratransgenesis are considered.
Mathematical modelling to assess the feasibility of Wolbachia in malaria vector biocontrol
22650S. Andreychuk and L. Yakob, Journal of Theoretical Biology, 542. 2022-03-29 06:30:06.
Releasing mosquitoes transinfected with the endosymbiotic bacterium Wolbachia is a novel strategy for interrupting vector-borne pathogen transmission. Following its success in controlling arboviruses spread by Aedes aegypti, this technology is being adapted for anopheline malaria vectors. However, antagonistic interactions between Wolbachia and naturally resident Asaia bacteria in malaria vectors have been demonstrated experimentally, potentially jeopardising Wolbachia biocontrol. We developed the first mathematical model accounting for interspecific competition between endosymbionts to assess the feasibility of this novel strategy for controlling malaria. First, Asaia prevalences among natural mosquito populations were compared with simulations parametrized with rates of Asaia transmission reported from laboratory studies. Discrepancies between projections and natural Asaia prevalences indicated potential overestimation of Asaia transmissibility in artificial laboratory settings. With parametrization that matches natural Asaia prevalence, simulations identified redundancies in Asaia's many infection routes (vertical, sexual and environmental). This resilience was only overcome when Wolbachia conferred very high resistance to environmental infection with Asaia, resulting in Wolbachia fixation and Asaia exclusion. Wolbachia's simulated spread was prevented when its maternal transmission was impeded in coinfected mosquitoes and the pre-control Asaia prevalence was beyond a threshold of 60-75%. This theoretical assessment highlights critical next steps in laboratory experiments to inform this strategy's feasibility. (c) 2022 The Author(s). Published by Elsevier Ltd.
Symbionts and gene drive: two strategies to combat vector-borne disease
21029G.-H. Wang, J. Du, C. Y. Chu, M. Madhav, G. L. Hughes and J. Champer, Trends in Genetics, 2022-03-18 07:56:11.
Mosquitoes bring global health problems by transmitting parasites and viruses such as malaria and dengue. Unfortunately, current insecticide-based control strategies are only moderately effective because of high cost and resistance. Thus, scalable, sustainable, and cost-effective strategies are needed for mosquito-borne disease control. Symbiont-based and genome engineering-based approaches provide new tools that show promise for meeting these criteria, enabling modification or suppression approaches. Symbiotic bacteria like Wolbachia are maternally inherited and manipulate mosquito host reproduction to enhance their vertical transmission. Genome engineering-based gene drive methods, in which mosquitoes are genetically altered to spread drive alleles throughout wild populations, are also proving to be a potentially powerful approach in the laboratory. Here, we review the latest developments in both symbionts and gene drive-based methods. We describe some notable similarities, as well as distinctions and obstacles, relating to these promising technologies.
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.
Quality over quantity: unraveling the contributions to cytoplasmic incompatibility caused by two coinfecting Cardinium symbionts
20317M. R. Doremus, C. M. Stouthamer, S. E. Kelly, S. Schmitz-Esser and M. S. Hunter, Heredity, 2022-02-05 09:08:33.
Cytoplasmic incompatibility (CI) is a common form of reproductive sabotage caused by maternally inherited bacterial symbionts of arthropods. CI is a two-step manipulation: first, the symbiont modifies sperm in male hosts which results in the death of fertilized, uninfected embryos. Second, when females are infected with a compatible strain, the symbiont reverses sperm modification in the fertilized egg, allowing offspring of infected females to survive and spread the symbiont to high frequencies in a population. Although CI plays a role in arthropod evolution, the mechanism of CI is unknown for many symbionts. Cardinium hertigii is a common CI-inducing symbiont of arthropods, including parasitoid wasps like Encarsia partenopea. This wasp harbors two Cardinium strains, cEina2 and cEina3, and exhibits strong CI. The strains infect wasps at different densities, with the cEina3 present at a lower density than cEina2, and it was previously not known which strain caused CI. By differentially curing wasps of cEina3, we found that this low-density symbiont is responsible for CI and modifies males during their pupal stage. cEina2 does not modify host reproduction and may spread by ‘hitchhiking’ with cEina3 CI or by conferring an unknown benefit. The cEina3 strain also shows a unique localization pattern in male reproductive tissues. Instead of infecting sperm like other CI-inducing symbionts, cEina3 cells are found in somatic cells at the testis base and around the seminal vesicle. This may allow the low-density cEina3 to efficiently modify host males and suggests that cEina3 uses a different modification strategy than sperm-infecting CI symbionts.
Recently introduced Wolbachia reduces bacterial species richness and reshapes bacterial community structure in Nilaparvata lugens
20141T.-P. Li, C.-Y. Zhou, J.-T. Gong, Z. Xi and X.-Y. Hong, Pest Management Science, 2022-01-21 13:28:22.
BACKGROUND Wolbachia has been developed as an effective tool to suppress insect pests and arbovirus transmission. Recently, the brown planthopper Nilaparvata lugens, a serious agricultural pest, has been successfully transinfected with Wolbachia strain wStri from Laodelphax striatellus. However, before conducting the field experiments, the impacts of wStri on the bacterial microbiota in N. lugens and how it differs from native Wolbachia wLug strain have not been clarified. RESULTS Here, we found that wStri reduced bacterial diversity and shaped bacterial community structure more than wLug in both developmental stage and different adult tissues. Overall, the relative abundance of Wolbachia was negatively correlated with bacterial diversity, but the bacterial diversity gradually decreased only when the relative abundance of Wolbachia was higher than 60%. Further analysis found that wStri reduced species richness of other bacteria but not their evenness. wStri infection also affected many bacterial functions (e.g., amino acid metabolism & signaling and cellular processes) in the developmental stages, with a stronger effect than wLug in nymphs. Moreover, although Wolbachia occupied a high relative abundance in infected individuals, Acinetobacter was consistently a core part of microbiome. CONCLUSION These results showed the significant impacts of recently introduced wStri on bacterial microbiota in N. lugens, with the effects differing from native wLug. This study will aid in understanding the relationship between Wolbachia, its host and the host's microbiota, and provide a reference for future field experiments. This article is protected by copyright. All rights reserved.
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.
Positive selection and horizontal gene transfer in the genome of a male-killing Wolbachia
19006T. Hill, R. L. Unckless and J. I. Perlmutter, Molecular Biology and Evolution, 2021-10-18 15:21:59.
Wolbachia are a genus of widespread bacterial endosymbionts in which some strains can hijack or manipulate arthropod host reproduction. Male killing is one such manipulation in which these maternally transmitted bacteria benefit surviving daughters in part by removing competition with the sons for scarce resources. Despite previous findings of interesting genome features of microbial sex ratio distorters, the population genomics of male-killers remain largely uncharacterized. Here, we uncover several unique features of the genome and population genomics of four Arizonan populations of a male-killing Wolbachia strain, wInn, that infects mushroom-feeding Drosophila innubila. We first compared the wInn genome to other closely related Wolbachia genomes of Drosophila hosts in terms of genome content and confirm that the wInn genome is largely similar in overall gene content to the wMel strain infecting D. melanogaster. However, it also contains many unique genes and repetitive genetic elements that indicate lateral gene transfers between wInn and non-Drosophila eukaryotes. We also find that, in line with literature precedent, genes in the Wolbachia prophage and Octomom regions are under positive selection. Of all the genes under positive selection, many also show evidence of recent horizontal transfer among Wolbachia symbiont genomes. These dynamics of selection and horizontal gene transfer across the genomes of several Wolbachia strains and diverse host species may be important underlying factors in Wolbachia’s success as a male-killer of divergent host species.
Microsporidia MB is found predominantly associated with Anopheles gambiae s.s and Anopheles coluzzii in Ghana
18931J. Akorli, E. A. Akorli, S. N. A. Tetteh, G. K. Amlalo, M. Opoku, R. Pwalia, M. Adimazoya, D. Atibilla, S. Pi-Bansa, J. Chabi and S. K. Dadzie, Scientific Reports, 11:5. 2021-09-20 19:43:33.
A vertically transmitted microsporidian, Microsporidia MB, with the ability to disrupt Plasmodium development was reported in Anopheles arabiensis from Kenya, East Africa. To demonstrate its range of incidence, archived DNA samples from 7575 Anopheles mosquitoes collected from Ghana were screened. MB prevalence was observed at 1.8%. An. gambiae s.s constituted 87% of positive mosquitoes while the remaining were from An. coluzzii. Both sibling species had similar positivity rates (24% and 19%; p = 0.42) despite the significantly higher number of An. gambiae s.s analysed (An. gambiae s.s = 487; An. coluzzii = 94; p = 0.0005). The microsporidian was also more prevalent in emerged adults from field-collected larvae than field-caught adults (p < 0.0001) suggestive of an efficient vertical transmission and/or horizontal transfer among larvae. This is the first report of Microsporidia MB in Anopheles mosquitoes in West Africa. It indicates possible widespread among malaria vector species and warrants investigations into the symbiont's diversity across sub-Saharan Africa.
Two newly introduced Wolbachia endosymbionts induce cell host differences in competitiveness and metabolic responses
18510T. P. Li, S. S. Zha, C. Y. Zhou, X. Xia, A. A. Hoffmann and X. Y. Hong, Appl Environ Microbiol, Aem0147921. 2021-09-09 14:34:33.
Wolbachia endosymbionts can induce multiple reproductive manipulations in their hosts, with cytoplasmic incompatibility (CI) being one of the most common manipulations. The important agricultural pests, white-backed planthopper (Sogatella furcifera) and brown planthopper (Nilaparvata lugens), are usually infected with CI-inducing Wolbachia wFur and non-CI-inducing Wolbachia wLug, respectively. The biological effects of these infections when present in a host cell are unknown. Here, we introduced the two Wolbachia strains into an Aedes albopictus cell line to stably establish a wFur-infected cell line (WFI) and a wLug-infected cell line (WLI). In a mixed culture, WFI cells were completely replaced by WLI cells, pointing to a stronger competitiveness of the WLI cell line. We found that infection by both Wolbachia strains reduced cell growth rates, but WLI had a faster cell growth rate than WFI, and this difference in cell growth rate combined with possible Wolbachia differences in diffusivity may have affected cell competitiveness. By examining gene expression and metabolites in the two lines, we found that some genes and key metabolites responded to differences in cell competitiveness. These results point to potential mechanisms that could contribute to the relative performance of hosts infected by these strains and also highlight the substantial impact of a non-CI Wolbachia on metabolism, which may in turn influence fitness of its native host. IMPORTANCE Wolbachia transinfection in insects can be used to suppress pests and block virus transmission. We stably introduced two Wolbachia strains from rice planthoppers into cell lines of an important arbovirus mosquito vector, Aedes albopictus. The competitiveness of host cells from the lines infected by the two Wolbachia strains was different, as were metabolic responses of the cell lines. These results suggest potential metabolic effects of Wolbachia on native hosts which could be exploited when they are transinfected into novel hosts for pest control.
Breakthrough in non-GMO malaria control
17914C. Robinson and J. Matthews, GM Watch, 2021-07-29 15:27:47.
A just-published study carried out in a high-security lab claims to show that a CRISPR gene drive (a way of forcing a heritable genetic modification through a whole species or population) can crash populations of malaria-spreading mosquitoes. But why crash mosquito populations with a risky experimental technology if you can completely stop them spreading malaria naturally? A naturally occurring biocontrol agent – a microbe – that inhibits the development of the malaria parasite in the mosquito Anopheles arabiensis, which spreads malaria in Sub-Saharan Africa, has recently been reported in the journal Nature Communications. Among the notable features of this approach are: • The microbe doesn’t seem to harm the mosquitoes in any way • Because it doesn’t kill the mosquitoes or cut their numbers, it should not have an impact on ecosystems dependent on them for food • The microbe seems to give the mosquitoes lifelong protection from malaria infection. The researchers are based at the International Centre of Insect Physiology and Ecology (ICIPE), Kenya, and in the UK. The microbe featured in their recent publication is a microsporidian. Microsporidia are unicellular spore-forming parasites that are now recognised as fungi, or as being related to fungi. All major animal groups harbour them, particularly insects. They spread horizontally, via spores that are ingested by a new host, but many also undergo vertical transmission to the next generation, via infected eggs (known as transovarial transmission).
Horizontal Transmission of the Symbiont Microsporidia MB in Anopheles arabiensis
17856G. Nattoh, T. Maina, E. E. Makhulu, L. Mbaisi, E. Mararo, F. G. Otieno, T. Bukhari, T. O. Onchuru, E. Teal, J. Paredes, J. L. Bargul, D. M. Mburu, E. A. Onyango, G. Magoma, S. P. Sinkins and J. K. Herren, Frontiers in Microbiology, 12. 2021-07-28 13:43:29.
The recently discovered Anopheles symbiont, Microsporidia MB, has a strong malaria transmission-blocking phenotype in Anopheles arabiensis, the predominant Anopheles gambiae species complex member in many active transmission areas in eastern Africa. The ability of Microsporidia MB to block Plasmodium transmission together with vertical transmission and avirulence makes it a candidate for the development of a symbiont-based malaria transmission blocking strategy. We investigate the characteristics and efficiencies of Microsporidia MB transmission between An. arabiensis mosquitoes. We show that Microsporidia MB is not transmitted between larvae but is effectively transmitted horizontally between adult mosquitoes. Notably, Microsporidia MB was only found to be transmitted between male and female An. arabiensis, suggesting sexual horizontal transmission. In addition, Microsporidia MB cells were observed infecting the An. arabiensis ejaculatory duct. Female An. arabiensis that acquire Microsporidia MB horizontally are able to transmit the symbiont vertically to their offspring. We also investigate the possibility that Microsporidia MB can infect alternate hosts that live in the same habitats as their An. arabiensis hosts, but find no other non-anopheline hosts. Notably, Microsporidia MB infections were found in another primary malaria African vector, Anopheles funestus s.s. The finding that Microsporidia MB can be transmitted horizontally is relevant for the development of dissemination strategies to control malaria that are based on the targeted release of Microsporidia MB infected Anopheles mosquitoes.
A natural symbiotic bacterium drives mosquito refractoriness to Plasmodium infection via secretion of an antimalarial lipase
17027H. Gao, L. Bai, Y. M. Jiang, W. Huang, L. L. Wang, S. G. Li, G. D. Zhu, D. Q. Wang, Z. H. Huang, X. S. Li, J. Cao, L. B. Jiang, M. Jacobs-Lorena, S. Zhan and S. B. Wang, Nature Microbiology, 25. 2021-05-06 11:00:53.
The stalling global progress in the fight against malaria prompts the urgent need to develop new intervention strategies. Whilst engineered symbiotic bacteria have been shown to confer mosquito resistance to parasite infection, a major challenge for field implementation is to address regulatory concerns. Here, we report the identification of a Plasmodium-blocking symbiotic bacterium, Serratia ureilytica Su_YN1, isolated from the midgut of wild Anopheles sinensis in China that inhibits malaria parasites via secretion of an antimalarial lipase. Analysis of Plasmodium vivax epidemic data indicates that local malaria cases in Tengchong (Yunnan province, China) are significantly lower than imported cases and importantly, that the local vector A. sinensis is more resistant to infection by P. vivax than A. sinensis from other regions. Analysis of the gut symbiotic bacteria of mosquitoes from Yunnan province led to the identification of S. ureilytica Su_YN1. This bacterium renders mosquitoes resistant to infection by the human parasite Plasmodium falciparum or the rodent parasite Plasmodium berghei via secretion of a lipase that selectively kills parasites at various stages. Importantly, Su_YN1 rapidly disseminates through mosquito populations by vertical and horizontal transmission, providing a potential tool for blocking malaria transmission in the field. The symbiotic bacterium Serratia ureilytica can disseminate through mosquito populations and render mosquitoes resistant to Plasmodium infection by secreting an antimalarial lipase.
Engineering a Culturable Serratia symbiotica Strain for Aphid Paratransgenesis
19389K. M. Elston, J. Perreau, G. P. Maeda, N. A. Moran and J. E. Barrick, Applied Environmental Microbiology, 87. 2020-12-06 17:18:42.
Aphids are global agricultural pests and important models for bacterial symbiosis. To date, none of the native symbionts of aphids have been genetically manipulated, which limits our understanding of how they interact with their hosts. Serratia symbiotica CWBI-2.3(T) is a culturable, gut-associated bacterium isolated from the black bean aphid. Closely related Serratia symbiotica strains are facultative aphid endosymbionts that are vertically transmitted from mother to offspring during embryogenesis. We demonstrate that CWBI-2.3(T) can be genetically engineered using a variety of techniques, plasmids, and gene expression parts. Then, we use fluorescent protein expression to track the dynamics with which CWBI-2.3(T) colonizes the guts of multiple aphid species, and we measure how this bacterium affects aphid fitness. Finally, we show that we can induce heterologous gene expression from engineered CWBI-2.3(T) in living aphids. These results inform the development of CWBI-2.3(T) for aphid paratransgenesis, which could be used to study aphid biology and enable future agricultural technologies.IMPORTANCE Insects have remarkably diverse and integral roles in global ecosystems. Many harbor symbiotic bacteria, but very few of these bacteria have been genetically engineered. Aphids are major agricultural pests and an important model system for the study of symbiosis. This work describes methods for engineering a culturable aphid symbiont, Serratia symbiotica CWBI-2.3(T) These approaches and genetic tools could be used in the future to implement new paradigms for the biological study and control of aphids.
Microbiome Innovation in Agriculture: Development of Microbial Based Tools for Insect Pest Management
19716M. Qadri, S. Short, K. Gast, J. Hernandez and A. C.-N. Wong, Frontiers in Sustainable Food Systems, 4. 2020-10-06 12:11:19.
This review emphasizes the potential and use of microbes in sustainable insect pest management. We first review the diverse insect traits shaped by insect-microbe associations that span nutrition, immunity, ecological interactions with natural enemy, insecticide resistance, and behavior. This is followed by discussing different microbiome manipulation approaches to alter pest traits, describing some of the opportunities and obstacles for each approach. We then highlight microbiomes as untapped chemical inventories to discover novel biopesticides, including plant-incorporated protectants and semiochemicals. The last topic covered is the use of beneficial microbes to improve mass-reared insects' performance for autocidal programs, including sterile insect technique and incompatible insect technique, in which we identify topics where data are limited or inconclusive, for future research.
The Evolving Arsenal Against Mosquito-Born Diseases
14369J. Smith, Labiotech.eu, 2020-09-10 16:01:02.
As the global climate continues to warm, disease-spreading mosquitoes such as Aedes aegypti are expected to establish themselves in the US and Europe.
Prospects and Pitfalls: Next-Generation Tools to Control Mosquito-Transmitted Disease
14366E. P. Caragata, S. Dong, Y. Dong, M. L. Simões, C. V. Tikhe and G. Dimopoulos, Annual Review of Microbiology, 74:455-475. 2020-09-08 15:57:19.
A diverse array of next-generation tools has been designed to eliminate mosquito populations or to replace them with mosquitoes that are less capable of transmitting key pathogens.
Engineered symbionts activate honey bee immunity and limit pathogens
19387P. Leonard Sean, J. E. Powell, J. Perutka, P. Geng, C. Heckmann Luke, D. Horak Richard, W. Davies Bryan, D. Ellington Andrew, E. Barrick Jeffrey and A. Moran Nancy, Science, 367:573-576. 2020-01-31 17:14:03.
Honey bees are essential pollinators threatened by colony losses linked to the spread of parasites and pathogens. Here, we report a new approach for manipulating bee gene expression and protecting bee health. We engineered a symbiotic bee gut bacterium, Snodgrassella alvi, to induce eukaryotic RNA interference (RNAi) immune responses. We show that engineered S. alvi can stably recolonize bees and produce double-stranded RNA to activate RNAi and repress host gene expression, thereby altering bee physiology, behavior, and growth. We used this approach to improve bee survival after a viral challenge, and we show that engineered S. alvi can kill parasitic Varroa mites by triggering the mite RNAi response. This symbiont-mediated RNAi approach is a tool for studying bee functional genomics and potentially for safeguarding bee health.
A bacterial gene-drive system efficiently edits and inactivates a high copy number antibiotic resistance locus
5595Valderrama, J. A., S. S. Kulkarni, V. Nizet and E. Bier, Nature Communications, 10:5726. 2019-12-17 15:48:28.
Gene-drive systems in diploid organisms bias the inheritance of one allele over another. CRISPR-based gene-drive expresses a guide RNA (gRNA) into the genome at the site where the gRNA directs Cas9-mediated cleavage. In the presence of Cas9, the gRNA cassette and any linked cargo sequences are copied via homology-directed repair (HDR) onto the homologous chromosome. Here, we develop an analogous CRISPR-based gene-drive system for the bacterium Escherichia coli that efficiently copies a gRNA cassette and adjacent cargo flanked with sequences homologous to the targeted gRNA/Cas9 cleavage site. This “pro-active” genetic system (Pro-AG) functionally inactivates an antibiotic resistance marker on a high copy number plasmid with ~ 100-fold greater efficiency than control CRISPR-based methods, suggesting an amplifying positive feedback loop due to increasing gRNA dosage. Pro-AG can likewise effectively edit large plasmids or single-copy genomic targets or introduce functional genes, foreshadowing potential applications to biotechnology or biomedicine.
New CRISPR system efficiently battles antibiotic resistance
5673Black, Samantha, The Science Advisory Board, 2019-12-16 19:02:07.
Researchers from the University of California San Diego have developed a brand new CRISPR-based gene-drive system that dramatically increases the efficiency of inactivation of genes responsible for antibiotic resistance. The new system is detailed on December 16 in Nature Communications. Genes conferring antibiotic resistance are often found on plasmids, circular forms of DNA that can replicate independently of the bacterial genome. Amplification effects of these plasmids can lead to the transfer of antibiotic resistance among bacteria. This poses a significant challenge to disrupt this function. Researchers have developed several cut-and-destroy methods but have had only moderate success with them.
Antibiotic resistance hoops countered by gene drive feedback loops
5670GEN, Genetic Engineering & Biotechnology News, 2019-12-16 18:56:57.
Where a gene drive that cuts and destroys plasmids may fail, a gene drive that cuts, pastes, and copies plasmids may succeed. The “where,” in this case, is an antibiotic-resistant bacterium that carries multiple copies of an antibiotic-resistance gene. That is, the target gene occurs on multiple copies of an “amplified” plasmid. If some plasmids remain intact, the antibiotic-resistance gene persists and may even be passed to other bacteria. To overcome this problem, scientists based at the University of California (UC), San Diego, developed a gene drive that incorporates a self-amplifying mechanism. Called Pro-AG, for “proactive” active genetics, it increases its efficiency through a positive feedback loop.
Tackling antibiotic resistance head-on with CRISPR
5937Thomas, L., News Medical Life Sciences, 2019-12-16 17:58:59.
With modern advances in genetic engineering occurring almost every day, the latest discovery concerns antibiotic resistance. Using the powerful gene editor CRISPR, scientists reported the development of a gene-drive system that is 100 times as efficient as other current systems at inactivating a specific bacterial gene responsible for making the bacterium antibiotic-resistant and which is present as multiple copies within the same bacterial cell. The paper, published on December 16, 2019, in the journal Nature Communications, uses the technology called active genetics, pioneered by biologists at UC San Diego.
Predicting the spatial dynamics of Wolbachia infections in Aedes aegypti arbovirus vector populations in heterogeneous landscapes
3917Hancock, PAR, S. A.; Koenraadt, C. J. M.; Scott, T. W.; Hoffmann, A. A.; Godfray, H. C. J., Journal of Applied Ecology, 56:1674-1686. 2019-01-15 00:00:00.
A promising strategy for reducing the transmission of dengue and other arboviral human diseases by Aedes aegypti mosquito vector populations involves field introductions of the endosymbiotic bacteria Wolbachia. Wolbachia infections inhibit viral transmission by the mosquito, and can spread between mosquito hosts to reach high frequencies in the vector population. Wolbachia spreads by maternal transmission, and spread dynamics can be variable and highly dependent on natural mosquito population dynamics, population structure and fitness components. We develop a mathematical model of an A. aegypti metapopulation that incorporates empirically validated relationships describing density-dependent mosquito fitness components. We assume that density dependent relationships differ across subpopulations, and construct heterogeneous landscapes for which model-predicted patterns of variation in mosquito abundance and demography approximate those observed in field populations. We then simulate Wolbachia release strategies similar to that used in field trials. We show that our model can produce rates of spatial spread of Wolbachia similar to those observed following field releases. We then investigate how different types of spatio-temporal variation in mosquito habitat, as well as different fitness costs incurred by Wolbachia on the mosquito host, influence predicted spread rates. We find that fitness costs reduce spread rates more strongly when the habitat landscape varies temporally due to stochastic and seasonal processes. Synthesis and applications: Our empirically based modelling approach represents effects of environmental heterogeneity on the spatial spread of Wolbachia. The models can assist in interpreting observed spread patterns following field releases and in designing suitable release strategies for targeting spatially heterogeneous vector populations.
Design, execution, and analysis of CRISPR-Cas9-based deletions and genetic interaction networks in the fungal pathogen Candida albicans
3916Halder, VP, C. B. M.; Chavez, A.; Shapiro, R. S., Nature Protocols, 14:955-975. 2019-01-14 00:00:00.
The study of fungal pathogens is of immediate importance, yet progress is hindered by the technical challenges of genetic manipulation. For Candida species, their inability to maintain plasmids, unusual codon usage, and inefficient homologous recombination are among the obstacles limiting efficient genetic manipulation. New advances in genomic biotechnologies -particularly CRISPR-based tools-have revolutionized genome editing for many fungal species. Here, we present a protocol for CRISPR-Cas9-based manipulation in Candida albicans using a modified gene-drive-based strategy that takes similar to 1 month to complete. We detail the generation of Candida-optimized Cas9-based plasmids for gene deletion, an efficient transformation protocol using C. albicans haploids, and an optimized mating strategy to generate homozygous single- and double-gene diploid mutants. We further describe protocols for quantifying cell growth and analysis pipelines to calculate fitness and genetic interaction scores for genetic mutants. This protocol overcomes previous limitations associated with genetic manipulation in C. albicans and advances researchers' ability to perform genetic analysis in this pathogen; the protocol also has broad applicability to other mating-competent microorganisms.
CRISPR in Parasitology: Not Exactly Cut and Dried!
3894Bryant, JMB, S.; Glover, L.; Hutchinson, S.; Rachidi, N., Trends in Parasitology, 35:409-422. 2019-01-12 00:00:00.
CRISPR/Cas9 technology has been developing rapidly in the field of parasitology, allowing for the dissection of molecular processes with unprecedented efficiency. Optimization and implementation of a new technology like CRISPR, especially in nonmodel organisms, requires communication and collaboration throughout the field. Recently, a 'CRISPR in Parasitology' symposium was held at the Institut Pasteur Paris, bringing together scientists studying Leishmania, Plasmodium, Trypanosoma, and Anopheles. Here we share technological advances and challenges in using CRISPR/Cas9 in the parasite and vector systems that were discussed. As CRISPR/Cas9 continues to be applied to diverse parasite systems, the community should now focus on improvement and standardization of the technique as well as expanding the CRISPR toolkit to include Cas9 alternatives/derivatives for more advanced applications like genome-wide functional screens.
Two-By-One model of cytoplasmic incompatibility: Synthetic recapitulation by transgenic expression of cifA and cifB in Drosophila
3944Shropshire, JDB, S. R., PLOS Genetics, 15:e1008221. 2019-01-02 00:00:00.
Wolbachia are maternally inherited bacteria that infect arthropod species worldwide and are deployed in vector control to curb arboviral spread using cytoplasmic incompatibility (CI). CI kills embryos when an infected male mates with an uninfected female, but the lethality is rescued if the female and her embryos are likewise infected. Two phage WO genes, cifA(wMel) and cifB(wMel) from the wMel Wolbachia deployed in vector control, transgenically recapitulate variably penetrant CI, and one of the same genes, cifA(wMel), rescues wild type CI. The proposed Two-by-One genetic model predicts that CI and rescue can be recapitulated by transgenic expression alone and that dual cifA(wMel) and cifB(wMel) expression can recapitulate strong CI. Here, we use hatch rate and gene expression analyses in transgenic Drosophila melanogaster to demonstrate that CI and rescue can be synthetically recapitulated in full, and strong, transgenic CI comparable to wild type CI is achievable. These data explicitly validate the Two-by-One model in wMel-infected D. melanogaster, establish a robust system for transgenic studies of CI in a model system, and represent the first case of completely engineering male and female animal reproduction to depend upon bacteriophage gene products. Author summary Releases of Wolbachia-infected mosquitos are underway worldwide because Wolbachia block replication of Zika and Dengue viruses and spread themselves maternally through arthropod populations via cytoplasmic incompatibility (CI). The CI drive system depends on a Wolbachia-induced sperm modification that results in embryonic lethality when an infected male mates with an uninfected female, but this lethality is rescued when the female and her embryos are likewise infected. We recently reported that the phage WO genes, cifA and cifB, cause the sperm modification and cifA rescues the embryonic lethality caused by the wMel Wolbachia strain deployed in vector control. These reports motivated proposal of the Two-by-One model of CI whereby two genes cause lethality and one gene rescues it. Here we provide unequivocal support for the model in the Wolbachia strain used in vector control via synthetic methods that recapitulate CI and rescue in the absence of a Wolbachia infections. Our results reveal the set of phage WO genes responsible for this powerful genetic drive system, act as a proof-of-concept that these genes alone can induce gene drive like crossing patterns, and establish methodologies and hypotheses for future studies of CI in Drosophila. We discuss the implications of the Two-by-One model towards functional mechanisms of CI, the emergence of incompatibility between Wolbachia strains, vector control applications, and CI gene nomenclature.
Engineered integrative and conjugative elements for efficient and inducible DNA transfer to undomesticated bacteria
3960Brophy, JANT, Alexander J.; Adams, Bryn L.; Renberg, Rebecca L.; Stratis-Cullum, Dimitra N.; Grossman, Alan D.; Voigt, Christopher A., Nature Microbiology, 3:1043-1053. 2018-01-18 00:00:00.
Engineering microorganisms to promote human or plant health will require manipulation of robust bacteria that are capable of surviving in harsh, competitive environments. Genetic engineering of undomesticated bacteria can be limited by an inability to transfer DNA into the cell. Here we developed an approach based on the integrative and conjugative element from Bacillus subtilis (ICEBs1) to overcome this problem. A donor strain (XPORT) was built to transfer miniaturized integrative and conjugative elements (mini-ICEBs1) to undomesticated bacteria. The strain was engineered to enable inducible control over conjugation, to integrate delivered DNA into the chromosome of the recipient, to restrict spread of heterologous DNA through separation of the type IV secretion system from the transferred DNA, and to enable simple isolation of engineered bacteria through a d-alanine auxotrophy. Efficient DNA transfer (10–1 to 10–7 conjugation events per donor) is demonstrated using 35 Gram-positive strains isolated from humans (skin and gut) and soil. Mini-ICEBs1 was used to rapidly characterize the performance of an isopropyl-?-d-thiogalactoside (IPTG)-inducible reporter across dozens of strains and to transfer nitrogen fixation to four Bacillus species. Finally, XPORT was introduced to soil to demonstrate DNA transfer under non-ideal conditions.
Rapid comeback of males: evolution of male-killer suppression in a green lacewing population
3979Hayashi, MN, M.; Kageyama, D., Proceedings of the Royal Society B-Biological Sciences, 285:6. 2018-01-17 00:00:00.
Evolutionary theory predicts that the spread of cytoplasmic sex ratio distorters leads to the evolution of host nuclear suppressors, although there are extremely few empirical observations of this phenomenon. Here, we demonstrate that a nuclear suppressor of a cytoplasmic male killer has spread rapidly in a population of the green lacewing Mallada desjardinsi. An M. desjardinsi population, which was strongly female-biased in 2011 because of a high prevalence of the male-killing Spiroplasma endosymbiont, had a sex ratio near parity in 2016, despite a consistent Spiroplasma prevalence. Most of the offspring derived from individuals collected in 2016 had 1 : 1 sex ratios in subsequent generations. Contrastingly, all-female or female-biased broods appeared frequently from crossings of these female offspring with males derived from a laboratory line founded by individuals collected in 2011. These results suggest near-fixation of a nuclear suppressor against male killing in 2016 and reject the notion that a non-male-killing Spiroplasma variant has spread in the population. Consistently, no significant difference was detected in mitochondrial haplotype variation between 2011 and 2016. These findings, and earlier findings in the butterfly Hypolimnas bolina in Samoa, suggest that these quick events of male recovery occur more commonly than is generally appreciated.
Development of a multi-locus CRISPR gene drive system in budding yeast
4029Yan, YF, Gregory C., Scientific reports, 8:17277-17277. 2018-01-07 00:00:00.
The discovery of CRISPR/Cas gene editing has allowed for major advances in many biomedical disciplines and basic research. One arrangement of this biotechnology, a nuclease-based gene drive, can rapidly deliver a genetic element through a given population and studies in fungi and metazoans have demonstrated the success of such a system. This methodology has the potential to control biological populations and contribute to eradication of insect-borne diseases, agricultural pests, and invasive species. However, there remain challenges in the design, optimization, and implementation of gene drives including concerns regarding biosafety, containment, and control/inhibition. Given the numerous gene drive arrangements possible, there is a growing need for more advanced designs. In this study, we use budding yeast to develop an artificial multi-locus gene drive system. Our minimal setup requires only a single copy of S. pyogenes Cas9 and three guide RNAs to propagate three gene drives. We demonstrate how this system could be used for targeted allele replacement of native genes and to suppress NHEJ repair systems by modifying DNA Ligase IV. A multi-locus gene drive configuration provides an expanded suite of options for complex attributes including pathway redundancy, combatting evolved resistance, and safeguards for control, inhibition, or reversal of drive action.
A transatlantic perspective on 20 emerging issues in biological engineering
4073Wintle, BCB, C. R.; Rhodes, C.; Molloy, J. C.; Millett, P.; Adam, L.; Breitling, R.; Carlson, R.; Casagrande, R.; Dando, M.; Doubleday, R.; Drexler, E.; Edwards, B.; Ellis, T.; Evans, N. G.; Hammond, R.; Haseloff, J.; Kahl, L.; Kuiken, T.; Lichman, B. R.; Matthewman, C. A.; Napier, J. A.; OhEigeartaigh, S. S.; Patron, N. J.; Perello, E.; Shapira, P.; Tait, J.; Takano, E.; Sutherland, W. J., eLife, 6:21. 2017-01-11 00:00:00.
Advances in biological engineering are likely to have substantial impacts on global society. To explore these potential impacts we ran a horizon scanning exercise to capture a range of perspectives on the opportunities and risks presented by biological engineering. We first identified 70 potential issues, and then used an iterative process to prioritise 20 issues that we considered to be emerging, to have potential global impact, and to be relatively unknown outside the field of biological engineering. The issues identified may be of interest to researchers, businesses and policy makers in sectors such as health, energy, agriculture and the environment.
Concept and history of genetic control
6014Scott, M. J. and Benedict, M. Q., Genetic Control of Malaria and Dengue, 2:31-54. 2016-12-30 20:16:45.
Genetic control of insects is an established method, mainly for insects that are important crop and veterinary pests such as medflies and screwworm. Efforts to use the same technologies against insects of medical importance, especially mosquitoes, have had limited success. The successes against mosquitoes have been accomplished using forms of both conventional and modern methods, both of which are promising. In this chapter, we provide highlights of the development of genetic control of agricultural pests and describe how the development of methods against mosquitoes reflects those advances. While admiring successful genetic control programs is motivating, we suggest that much can also be learned from both past successful and failed efforts, as doing so will increase our ability to improve future activities.
Contact Us
Alex Sullivan
Foundation for the
National Institutes of Health
geneconvenevi@fnih.org