Keywords: Cytoplasmic incompatibility
Dengue Suppression by Male Wolbachia-Infected Mosquitoes
35443Jue Tao Lim, Ph.D., Chee-Seng Chong, Ph.D., Chia-Chen Chang, Ph.D., et al., New England Journal of Medicine, 2026-02-12 17:46:04.
Wild-type female Aedes aegypti mosquitoes that mate with male A. aegypti mosquitoes that have been infected with the wAlbB strain of Wolbachia pipientis bacteria produce nonviable offspring owing to cytoplasmic incompatibility. Repeated releases of wolbachia-infected males can potentially suppress wild-type mosquito populations and reduce the risk of dengue virus infection. We conducted a trial involving the release of male A. aegypti mosquitoes infected with the wAlbB strain of wolbachia bacteria for the control of dengue in Singapore, a tropical city-state. In this cluster-randomized trial with test-negative controls, we divided 15 geographic population clusters into two groups: 8 clusters received deployments of male wolbachia-infected mosquitoes (intervention clusters) and 7 clusters received no deployments (control clusters). The primary end point was the diagnosis of symptomatic dengue virus infection of any severity caused by any serotype of the virus, as measured by the odds ratio for the distribution of wolbachia exposure among laboratory-confirmed reported dengue cases as compared with test-negative controls. A total of 393,236 residents lived in the intervention clusters, and 331,192 lived in the control clusters. Adult wild-type A. aegypti populations were suppressed across the intervention clusters. The baseline average abundance of the mosquitoes (number of adult female mosquitoes trapped divided by number of traps) was 0.18 and 0.19 in the intervention and control clusters, respectively; from 3 months after the initiation of the intervention until the end of the 24-month trial period, the average abundance was 0.041 and 0.277, respectively. In the intention-to-treat analysis at 6 months or more, the percentage of residents in the intervention clusters who were dengue-positive was lower than that in the control clusters (354 of 5722 tests [6%] vs. 1519 of 7080 tests [21%]). The protective efficacy of the intervention, calculated as (1−odds ratio)×100, ranged from 71 to 72% with 3 to 12 months or more of wolbachia mosquito exposure, as represented by odds ratios of 0.28 to 0.29. Release of sterile wolbachia-infected male A. aegypti mosquitoes reduced vector populations and the risk of dengue infection in Singapore.
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.
Spatial Dynamics and Sterilization Range of Incompatible Aedes albopictus Males: Advancing Toward an Optimized IIT Approach
35474Lampazzi, E., Virgillito, C., Caputo, B., et al., Tropical Medicine and Infectious Disease, 11. 2026-02-06 18:17:05.
The Incompatible Insect Technique (IIT) is a species-specific, eco-friendly mosquito control method that relies on releasing Wolbachia-infected males, which induce cytoplasmic incompatibility (CI), rendering eggs inviable when mating with wild females. Aiming at optimizing IIT protocols in terms of cost-effectiveness, data on incompatible male dispersal and survival and the distance- and time-related impact of induced sterility are fundamental. This study plans to fill this gap and reports findings from a two-year field trial (2022–2023) at the ENEA-Casaccia Research Center, based on single-spot releases of incompatible Aedes albopictus males (ARwP strain). Male releases were carried out in late September 2022 (~15,000 released males) and the early Ae. albopictus season (at the end of June 2023; ~24,000 released males). Fifty-eight ovitraps were located at a 20–900 m distance from the ARwP release spot and were monitored weekly from May to November to assess egg hatching rates and measure CI effects in relation to both distance and time. Following the 2023 release, samples of adults were collected at increasing distances from the release site and at multiple post-release time points to assess, individually, wild female fertility and ARwP male dispersal and survival using Wolbachia as a genetic marker. Statistical analyses revealed that: (a) the highest reduction in the egg hatching was found within 100 m from the release spot (46.5% and 19.9%, respectively, in 2022 and 2023) but remained significant even at greater distances (29.9% and 7.7% at 300 m, respectively, in 2022 and 2023); (b) accordingly, the highest reduction in the wild female fertility occurred within 100 m from the release spot (47.3%), but similar effects were recognizable up to 600 m; (c) the overflooding ratio of the ARwP males did not significantly differ between 3 and 11 days after the release, with ARwP males remaining active up to 18 days and dispersing as far as 400 m. These results demonstrate the potential of localized, non-inundative IIT trials to furnish clues for the setup of spatially optimized release strategies, especially in scaled-up applications. The study also emphasizes the need for standardized assessment tools and further research regarding environmental and behavioral factors influencing long-term suppression outcomes.
Wolbachia-Based biocontrol of Aedes aegypti: Current Progress, Challenges, and future prospects
35217Ashif Ahamed, Safdar Ali, Mehboob Hoque, Journal of Invertebrate Pathology, 2025-10-05 13:03:30.
Wolbachia is used as potential bio-control tool to reduce the dengue mosquitoes and also to reduce dengue virus (DENV) load in transinfected Aedes aegypti. Different field trials including the ones by the World Mosquito Program (WMP) have deployed Wolbachia carrying DENV primary vectors at different target sites worldwide. Field trials suggest that through the Population Replacement Strategy (PRS) and Incompatible Insect Technique (IIT), Wolbachia can introgress into the local Ae. aegypti population or reduce the population size of the Aedes aegypti by cytoplasmic incompatibility phenomenon and subsequent reduction in dengue incidences at target sites. Challenges for this strategy are to establish and maintain the transinfected mosquito population, choosing the appropriate strain of the bacteria, long term establishment of transinfected dengue mosquitoes for several subsequent generations, selecting the appropriate approach of mosquito field release, operational challenges, and the viral evolution. Moreover, the outcome of Wolbachia mediated biological control of Aedes aegypti in a broad scale is yet to be seen and the long-term stability of Wolbachia in transinfected mosquitoes remains unconfirmed. Significant success has been recorded in a broad scale deployment of wAlbB-carrying Aedes aegypti through the ongoing Wolbachia Malaysia project, resulting in a reduction in dengue fever cases. Successful large-deployment of this strategy has also been performed in Colombia and Indonesia (1.7 to 3.3 million people and 135–540 km2 area). Similar investigations on broad scale for longer period across different environmental conditions using transinfected mosquito deployment and associated risk analysis are imperative before adopting this biological control approach as a potent dengue control strategy worldwide.
Genomic Insights into Wolbachia Strain wCin2USA1 Reveal Promising Cytoplasmic Incompatibility Potential and Next-Generation Dengue Biocontrol
35098Istiaque Zaeem, Nurnabi Azad Jewel, Mohimenul Haque Rolin, et al., bioRxiv, 2025-09-01 19:59:21.
Dengue fever is one of the fastest-growing mosquito-borne diseases in the world, causing illness and economic challenges in many countries. Current mosquito control methods, such as insecticides, are often costly, less effective over time, and harmful to the environment. An alternative approach uses naturally occurring bacteria called Wolbachia, which live inside insects. When certain Wolbachia strains are introduced into mosquitoes, they can reduce the insects’ ability to spread viruses like dengue. In our study, we compared the genetic makeup of several Wolbachia strains to identify new candidates that may work better for controlling dengue. We discovered that a strain called wCin2USA1 has strong potential because it shares important features with two strains already used successfully, while also having unique advantages. These include genes that help prevent mosquitoes from reproducing normally when carrying different Wolbachia strains, which can reduce mosquito populations. Our findings suggest that wCin2USA1 could be developed as a new, environmentally friendly tool to help reduce dengue transmission.
Global asymptotic stability in a delay stage structured model for mosquito population suppression
34720Huang, Mg., Yu, Js., Applied Mathematics, 40:112-136. 2025-03-31 14:42:25.
A promising avenue to control mosquito-borne diseases such as dengue, malaria, and Zika involves releasing male mosquitoes carrying the bacterium Wolbachia in wild areas to drive female sterility by a mechanism called cytoplasmic incompatibility (CI). In this work, we initiate a preliminary assessment of how the combined impact of dispersal, incomplete CI and mating competitiveness on mosquito population suppression by a delay differential equation model. Our theoretical analyses indicate that the immigration of eggs plays a significant role in the suppression dynamics. For the case without egg immigration, we identify a threshold dispersal rate v* of adult mosquitoes, threshold CI density ξ*, and threshold release ratio r*. A successful mosquito suppression would be established only when v < v*, ξ > ξ*, and r(t) ≥ r* uniformly. The immigration of eggs causes the threshold dynamics to be invalid, and warns an absolute failure of population suppression. The monotonicity of the adult steady-state in the dispersal rate and CI intensity indicates that choosing a suitable Wolbachia strain with strong CI intensity, or bringing down the dispersal rate of mosquitoes by blocking the suppression zones is a feasible strategy to obtain a better suppression level.
Exploiting Wolbachia as a Tool for Mosquito-Borne Disease Control: Pursuing Efficacy, Safety, and Sustainability
34677Moretti, R., Lim, J. T., Ferreira, A. G. A., et al, Pathogens, 14:285. 2025-03-28 09:41:16.
Despite the application of control measures, mosquito-borne diseases continue to pose a serious threat to human health. In this context, exploiting Wolbachia, a common symbiotic bacterium in insects, may offer effective solutions to suppress vectors or reduce their competence in transmitting several arboviruses. Many Wolbachia strains can induce conditional egg sterility, known as cytoplasmic incompatibility (CI), when infected males mate with females that do not harbor the same Wolbachia infection. Infected males can be mass-reared and then released to compete with wild males, reducing the likelihood of wild females encountering a fertile mate. Furthermore, certain Wolbachia strains can reduce the competence of mosquitoes to transmit several RNA viruses. Through CI, Wolbachia-infected individuals can spread within the population, leading to an increased frequency of mosquitoes with a reduced ability to transmit pathogens. Using artificial methods, Wolbachia can be horizontally transferred between species, allowing the establishment of various laboratory lines of mosquito vector species that, without any additional treatment, can produce sterilizing males or females with reduced vector competence, which can be used subsequently to replace wild populations. This manuscript reviews the current knowledge in this field, describing the different approaches and evaluating their efficacy, safety, and sustainability. Successes, challenges, and future perspectives are discussed in the context of the current spread of several arboviral diseases, the rise of insecticide resistance in mosquito populations, and the impact of climate change. In this context, we explore the necessity of coordinating efforts among all stakeholders to maximize disease control. We discuss how the involvement of diverse expertise—ranging from new biotechnologies to mechanistic modeling of eco-epidemiological interactions between hosts, vectors, Wolbachia, and pathogens—becomes increasingly crucial. This coordination is especially important in light of the added complexity introduced by Wolbachia and the ongoing challenges posed by global change.
wStri spread dynamics in Nilaparvata lugens via discrete mathematical models
34574Zheng, B., Yang, H., Elaydi, S. et al., Journal of Mathematical Biology, 90. 2025-03-12 16:30:16.
Wolbachia, an intracellular bacterium, is well-known for inducing cytoplasmic incompatibility, which has become a promising and environmentally sustainable strategy for controlling pest populations. The strain wStri, specifically identified in Nilaparvata lugens (brown planthopper), has shown potential for such biocontrol applications. In this study, we develop a comprehensive discrete mathematical model to analyze the dynamics of wStri spread in a mixed population of wStri-infected, wLug-infected, and uninfected Nilaparvata lugens under both constant and periodically varying environmental conditions. Under a constant environment, the model identifies the critical threshold necessary for the successful establishment of wStri within the population. Our analysis reveals that the model exhibits a strong Allee effect, where a population must exceed a certain critical density-the Allee threshold-for the wStri strain to persist and spread. Below this threshold, the wStri strain is likely to be eliminated, failing in pest control efforts. When the environment varies periodically, the model transforms into a non-autonomous periodic discrete model, introducing additional complexity. In this scenario, we derive sufficient conditions that ensure the composition of finitely many Allee maps continues to function as an Allee map. Furthermore, we prove that a unique periodic orbit exists within such a periodic environment. This orbit is characterized as unstable and acts as a threshold, determining whether wStri will establish itself in the population or die out over time. The findings from this model provide critical insights into the conditions under which wStri can be effectively used to control Nilaparvata lugens, particularly in environments that are not constant but fluctuate periodically. These insights have significant implications for the practical deployment of Wolbachia-based biocontrol methods in pest management strategies.
Wolbachia Invasion in Mosquitoes with Incomplete CI, Imperfect Maternal Transmission and Maturation Delay
32710Ma X, Su Y, Bulletin of Mathematical Biology, 2024-10-22 17:32:46.
The mechanism of cytoplasmic incompatibility (CI) is important in the study of Wolbachia invasion in wild mosquitoes. Su et al. (Bull Math Biol 84(9):95, 2022) proposed a delay differential equation model by relating the CI effect to maturation delay. In this paper, we investigate the dynamics of this model by allowing the same density-dependent death rate and distinct density-independent death rates. Through analyzing the existence and stability of equilibria, we obtain the parameter conditions for Wolbachia successful invasion if the maternal transmission is perfect. While if the maternal transmission is imperfect, we give the ranges of parameters to ensure failure invasion, successful invasion and partially suppressing, respectively. Meanwhile, numerical simulations indicate that the system may exhibit monostable and bistable dynamics when parameters vary. Particularly, in the bistable situation an unstable separatrix, like a line, exists when choosing constant functions as initial values; and the maturation delay affects this separatrix in an interesting way.
Stable introduction of Wolbachia wPip into invasive Anopheles stephensi for potential malaria control
32022Yongkang Liang, Julian Liu, et al., PLoS Neglected Tropical Diseases, 2024-10-01 16:23:02.
Recent efforts have successfully utilized the endosymbiotic bacterium Wolbachia to control mosquito-transmitted viral diseases like dengue in multiple countries. However, similar initiatives have been limited in combating malaria, the most devastating and deadly mosquito-borne disease, which claims over half a million lives each year. This is primarily due to the difficulty in establishing a stable, maternally inheritable Wolbachia infection in Anopheles, the mosquito vector responsible for malaria transmission. A significant concern in malaria control is the invasion of the urban malaria vector Anopheles stephensi into Africa, where malaria burden is highest, and over 40% of the population resides in urban areas. Building on the previous breakthrough of establishing Wolbachia strain wAlbB in An. stephensi, the author has now achieved a second stable infection by transferring the wPip strain from Culex pipiens into this mosquito species using embryonic microinjection. The resultant transinfected HP1 line induces nearly complete cytoplasmic incompatibility when crossed with wild mosquitoes, displays robust fitness and male mating competitiveness, and exhibits strong resilience against heat stress. These advantageous traits position the HP1 line as a promising candidate for further development in malaria control.
Wolbachia Infection through Hybridization to Enhance an Incompatible Insect Technique-Based Suppression of Aedes albopictus in Eastern Spain
29086Cholvi M, Trelis M, Bueno-Marí R, Khoubbane M, Gil R, Marcilla A, Moretti R., Insects, 15. 2024-04-04 09:29:30.
Wolbachia bacteria occur naturally as symbionts of many insect species and are responsible for various phenomena that modify the hosts’ reproductive biology. Among them, cytoplasmic incompatibility (CI) refers to the sterility of eggs produced by crosses between infected males and females that are uninfected or infected by a non-compatible strain of these bacteria. CI can be exploited for vector control by establishing an opportune Wolbachia infection in a laboratory population of a target insect species and then releasing the infected males into the environment as sterilizing agents. In the present work, a suitable Wolbachia strain was introduced into a Spanish population of the Asian tiger mosquito, Aedes albopictus, through hybridization with the laboratory line, ARwP, already tested as an efficient control tool against this vector. The obtained hybrids were compared with the ARwP to ascertain the effects derived from transferring the infection to a different Ae. albopictus population. No significant differences between lines were found regarding survival, female fecundity, and egg fertility. Importantly, the eggs produced by crosses between males of the hybrid lines and unmodified wild females were 99.9% sterile. This result encourages further studies to explore the feasibility of a Wolbachia-based control program against the Asian tiger mosquito in Spain.
Prophage proteins alter long noncoding RNA and DNA of developing sperm to induce a paternal-effect lethality
29033Rupinder Kaur et al., Science, 383:1111-1117. 2024-03-19 15:33:59.
Many arthropods carry symbiotic bacteria. Some, including Wolbachia strains, have the capacity to cause male sterility.
PSRs: Selfish chromosomes that manipulate reproductive development
28974Xinmi Zhang, Patrick M. Ferree, Seminars in Cell and Developmental Biology, 159-160:66-73. 2024-03-05 14:28:18.
B chromosomes are intriguing “selfish” genetic elements, many of which exhibit higher-than-Mendelian transmission. This perspective highlights a group of B chromosomes known as Paternal Sex Ratio chromosomes (PSRs), which are found in several insects with haplo-diploid reproduction. PSRs harshly alter the organism’s reproduction to facilitate their own inheritance. A manifestation of this effect is the conversion of female destined individuals into males. Key to this conversion is the mysterious ability of PSRs to cause elimination of the sperm-inherited half of the genome during zygote formation. Here we discuss how PSRs were discovered, what is known about how they alter paternal chromatin dynamics to cause sex conversion, and how PSR-induced genome elimination is different from other forms of programmed genome elimination in different insects. PSRs also stand out because their DNA sequence compositions differ in remarkable ways from their insect’s essential chromosomes, a characteristic suggestive of interspecies origins. Broadly, we also highlight poorly understood aspects of PSR dynamics that need to be investigated.
Modelling and analysis of periodic impulsive releases of the Nilaparvata lugens infected with wStri-Wolbachia
28638Xiangjun Dai, Qi Quan and Jianjun Jiao, Journal of Biological Dynamics, 17:1. 2023-12-04 12:23:12.
In this paper, we formulate a population suppression model and a population replacement model with periodic impulsive releases of Nilaparvata lugens infected with wStri. The conditions for the stability of wild-N.lugens -eradication periodic solution of two systems are obtained by applying the Floquet theorem and comparison theorem. And the sufficient conditions for the persistence in the mean of wild N.lugens are also given. In addition, the sufficient conditions for the extinction and persistence of the wild N.lugens in the subsystem without wLug are also obtained. Finally, we give numerical analysis which shows that increasing the release amount or decreasing the release period are beneficial for controlling the wild N.lugens, and the efficiency of population replacement strategy in controlling wild populations is higher than that of population suppression strategy under the same release conditions.
Wolbachia endosymbionts manipulate the self-renewal and differentiation of germline stem cells to reinforce fertility of their fruit fly host
28215S. L. Russell, J. R. Castillo and W. T. Sullivan, PLOS Biology, 21:e3002335. 2023-10-24 14:41:16.
The alphaproteobacterium Wolbachia pipientis infects arthropod and nematode species worldwide, making it a key target for host biological control. Wolbachia-driven host reproductive manipulations, such as cytoplasmic incompatibility (CI), are credited for catapulting these intracellular bacteria to high frequencies in host populations. Positive, perhaps mutualistic, reproductive manipulations also increase infection frequencies, but are not well understood. Here, we identify molecular and cellular mechanisms by which Wolbachia influences the molecularly distinct processes of germline stem cell (GSC) self-renewal and differentiation. We demonstrate that wMel infection rescues the fertility of flies lacking the translational regulator mei-P26and is sufficient to sustain infertile homozygous mei-P26knockdown stocks indefinitely. Cytology revealed that wMel mitigates the impact of mei-P26 loss through restoring proper pMad, Bam, Sxl, and Orb expression. In Oregon R files with wild-type fertility, wMel infection elevates lifetime egg hatch rates. Exploring these phenotypes through dual-RNAseq quantification of eukaryotic and bacterial transcripts revealed that wMel infection rescues and offsets many gene expression changes induced by meiP26loss at the mRNA level. Overall, we show that wMel infection beneficially reinforces host fertility at mRNA, protein, and phenotypic levels, and these mechanisms may promote the emergence of mutualism and the breakdown of host reproductive manipulations.
How mosquito-controlling bacteria might also enhance insect fertility
28212Public Library of Science, Phys Org, 2023-10-24 13:51:56.
A new study reveals biological mechanisms by which a specific strain of bacteria in the Wolbachia genus might enhance the fertility of the insects it infects—with potentially important implications for mosquito-control strategies. Shelbi Russell of the University of California Santa Cruz, US, and colleagues reported these findings in the open access journal PLOS Biology on October 24th. Different strains of Wolbachia bacteria naturally infect a number of different animals worldwide, such as mosquitos, butterflies, and fruit flies. Wolbachia can manipulate the fertility of their hosts through a specific biological mechanism that aids the spread of Wolbachia within host populations. In recent years, people have harnessed that mechanism in strategies to deliberately infect mosquitos with a specific Wolbachia strain, reducing targeted mosquito populations and thereby potentially reducing the spread of human viruses carried by mosquitos, such as dengue or Zika.
Wolbachia Induces Structural Defects Harmful to Drosophila simulans Riverside Spermiogenesis.
28142M. G. Riparbelli, A. Pratelli and G. Callaini, Cells, 12. 2023-09-21 10:31:00.
The relationship between cytoplasmic incompatibility and the obligate intracellular alphaproteobacteria Wolbachia has for a long time been reported. Although the molecular mechanisms responsible for this reproductive alteration are beginning to be understood, the effects of Wolbachia on germ cell structure and dynamics have not yet been fully investigated. We report here that the presence of Wolbachia in infected cysts of elongating spermatids is associated with major structural defects that become more evident in mature sperm. We find mitochondrial defects, an improper axoneme structure, reduced sperm numbers, and individualization failures. The large heterogeneous variety of the ultrastructural defects found in elongating spermatids and mature sperm provide the first cytological evidence for the reduced fertility associated with Wolbachia infection in Drosophila simulans males. The observed abnormalities could be the result of the mechanical stress induced by the high bacteria numbers during the process of spermatid elongation, rather than the result of the released factors affecting the proper morphogenesis of the germ cells. Moreover, high Wolbachia densities in male germ cells may not be appropriate for causing cytoplasmic incompatibility as the bacteria are harmful for spermatid differentiation, leading to abnormal sperm that is unlikely to be functional.
Wolbachia infection at least partially rescues the fertility and ovary defects of several new Drosophila melanogaster bag of marbles protein-coding mutants
27701W. Miwa and F. A. Charles, bioRxiv, 2023.03.20.532813. 2023-08-16 09:19:09.
The D. melanogaster protein coding gene bag of marbles (bam) plays a key role in early male and female reproduction by forming complexes with partner proteins to promote differentiation in gametogenesis. Like another germline gene, Sex lethal, bam genetically interacts with the endosymbiont Wolbachia, as Wolbachia rescues the reduced fertility of a bam hypomorphic mutant. Here, we explored the specificity of the bam-Wolbachia interaction by generating 22 new bam mutants, with ten mutants displaying fertility defects. Nine of these mutants trend towards rescue by the wMel Wolbachia variant, with eight statistically significant at the fertility and/or cytological level. In some cases, fertility was increased a striking 20-fold. There is no specificity between the rescue and the known binding regions of bam, suggesting wMel does not interact with one singular bam partner to rescue the reproductive phenotype. We further tested if wMel interacts with bam in a non-specific way, by increasing bam transcript levels or acting upstream in germline stem cells. A fertility assessment of a bam RNAi knockdown mutant reveals that wMel rescue is specific to functionally mutant bam alleles and we find no obvious evidence of wMel interaction with germline stem cells in bam mutants.Author Summary Reproduction in the Drosophila melanogaster fruit fly is dependent on the bag of marbles (bam) gene, which acts early in the process of generating eggs and sperm. Mutations to this gene negatively impact the fertility of the fly, causing it to be sterile or have fewer progeny. Interestingly, we find that the bacteria Wolbachia, which resides within reproductive cells across a wide range of insects, partially restores the fertility and ovary phenotype of several bam mutants of which the resultant Bam protein is altered from wildtype. The protein function of Bam is further suggested to be important by the lack of rescue for a fly that has a fertility defect due to low expression of a non-mutated bam gene. Previous work makes similar conclusions about Wolbachia with another reproductive gene, Sex lethal (Sxl), highlighting the potential for rescue of fertility mutants to occur in a similar way across different genes. An understanding of the ways in which Wolbachia can affect host reproduction provides us with context with which to frame Wolbachia’s impact on host genes, such as bam and Sxl, and consider the evolutionary implications of Wolbachia’s infection in D. melanogaster fruit flies.Competing Interest StatementThe authors have declared no competing interest.
First Detection and Genetic Identification of Wolbachia Endosymbiont in Field-Caught Aedes aegypti (Diptera: Culicidae) Mosquitoes Collected from Southern Taiwan
27713L.-L. Chao and C.-M. Shih, Microorganisms, 11. 2023-07-23 09:53:12.
The prevalence and genetic character of Wolbachia endosymbionts in field-collected Aedes aegypti mosquitoes were examined for the first time in Taiwan. A total of 665 Ae. aegypti were screened for Wolbachia infection using a PCR assay targeting the Wolbachia surface protein (wsp) gene. In general, the prevalence of Wolbachia infection was detected in 3.3% Ae. aegypti specimens (2.0% female and 5.2% male). Group-specific Wolbachia infection was detected with an infection rate of 1.8%, 0.8% and 0.8% in groups A, B and A&B, respectively. Genetic analysis demonstrated that all Wolbachia strains from Taiwan were phylogenetically affiliated with Wolbachia belonging to the supergroups A and B, with high sequence similarities of 99.4–100% and 99.2–100%, respectively. Phylogenetic relationships can be easily distinguished by maximum likelihood (ML) analysis and were congruent with the unweighted pair group with the arithmetic mean (UPGMA) method. The intra- and inter-group analysis of genetic distance (GD) values revealed a lower level within the Taiwan strains (GD < 0.006 for group A and GD < 0.008 for group B) and a higher level (GD > 0.498 for group A and GD > 0.286 for group B) as compared with other Wolbachia strains. Our results describe the first detection and molecular identification of Wolbachia endosymbiont in field-caught Ae. aegypti mosquitoes collected from Taiwan, and showed a low Wolbachia infection rate belonging to supergroups A and B in Ae. aegypti mosquitoes.
26619
N. Wybouw, E. Van Reempts, J. Zarka, F. Zélé and D. Bonte, Heredity, 2023-07-13 08:18:54.
Maternally transmitted symbionts such as Wolbachia can alter sex allocation in haplodiploid arthropods. By biasing population sex ratios towards females, these changes in sex allocation may facilitate the spread of symbionts. In contrast to symbiont-induced cytoplasmic incompatibility (CI), the mechanisms that underpin sex allocation distortion remain poorly understood. Using a nuclear genotype reference panel of the haplodiploid mite Tetranychus urticae and a single Wolbachia variant that is able to simultaneously induce sex allocation distortion and CI, we unraveled the mechanistic basis of Wolbachia-mediated sex allocation distortion. Host genotype was an important determinant for the strength of sex allocation distortion. We further show that sex allocation distortion by Wolbachia in haplodiploid mites is driven by increasing egg size, hereby promoting egg fertilization. This change in reproductive physiology was also coupled to increased male and female adult size. Our results echo previous work on Cardinium symbionts, suggesting that sex allocation distortion by regulating host investment in egg size is a common strategy among symbionts that infect haplodiploids. To better understand the relevance that sex allocation distortion may have for the spread of Wolbachia in natural haplodiploid populations, we parametrized a model based on generated phenotypic data. Our simulations show that empirically derived levels of sex allocation distortion can be sufficient to remove invasion thresholds, allowing CI to drive the spread of Wolbachia independently of the initial infection frequency. Our findings help elucidate the mechanisms that underlie the widespread occurrence of symbionts in haplodiploid arthropods and the evolution of sex allocation.
The cellular lives of Wolbachia
26608J. Porter and W. Sullivan, Nature Reviews Microbiology, 2023-07-10 13:14:07.
Wolbachia are successful Gram-negative bacterial endosymbionts, globally infecting a large fraction of arthropod species and filarial nematodes. Efficient vertical transmission, the capacity for horizontal transmission, manipulation of host reproduction and enhancement of host fitness can promote the spread both within and between species. Wolbachia are abundant and can occupy extraordinary diverse and evolutionary distant host species, suggesting that they have evolved to engage and manipulate highly conserved core cellular processes. Here, we review recent studies identifying Wolbachia–host interactions at the molecular and cellular levels. We explore how Wolbachia interact with a wide array of host cytoplasmic and nuclear components in order to thrive in a diversity of cell types and cellular environments. This endosymbiont has also evolved the ability to precisely target and manipulate specific phases of the host cell cycle. The remarkable diversity of cellular interactions distinguishes Wolbachia from other endosymbionts and is largely responsible for facilitating its global propagation through host populations. Finally, we describe how insights into Wolbachia–host cellular interactions have led to promising applications in controlling insect-borne and filarial nematode-based diseases.
Modeling emergence of Wolbachia toxin-antidote protein functions with an evolutionary algorithm
26277J. Beckmann, J. Gillespie and D. Tauritz, Front Microbiol, 14:1116766. 2023-06-09 10:36:38.
Evolutionary algorithms (EAs) simulate Darwinian evolution and adeptly mimic natural evolution. Most EA applications in biology encode high levels of abstraction in top-down population ecology models. In contrast, our research merges protein alignment algorithms from bioinformatics into codon based EAs that simulate molecular protein string evolution from the bottom up. We apply our EA to reconcile a problem in the field of Wolbachia induced cytoplasmic incompatibility (CI). Wolbachia is a microbial endosymbiont that lives inside insect cells. CI is conditional insect sterility that operates as a toxin antidote (TA) system. Although, CI exhibits complex phenotypes not fully explained under a single discrete model. We instantiate in-silico genes that control CI, CI factors (cifs), as strings within the EA chromosome. We monitor the evolution of their enzymatic activity, binding, and cellular localization by applying selective pressure on their primary amino acid strings. Our model helps rationalize why two distinct mechanisms of CI induction might coexist in nature. We find that nuclear localization signals (NLS) and Type IV secretion system signals (T4SS) are of low complexity and evolve fast, whereas binding interactions have intermediate complexity, and enzymatic activity is the most complex. Our model predicts that as ancestral TA systems evolve into eukaryotic CI systems, the placement of NLS or T4SS signals can stochastically vary, imparting effects that might impact CI induction mechanics. Our model highlights how preconditions and sequence length can bias evolution of cifs toward one mechanism or another.
Molecular Biology of Cytoplasmic Incompatibility Caused by Wolbachia Endosymbionts
25549M. Hochstrasser, Annual Review of Microbiology, 2023-06-07 08:38:45.
Among endosymbiotic bacteria living within eukaryotic cells, Wolbachia is exceptionally widespread, particularly in arthropods. Inherited through the female germline, it has evolved ways to increase the fraction of bacterially infected offspring by inducing parthenogenesis, feminization, male killing, or, most commonly, cytoplasmic incompatibility (CI). In CI, Wolbachia infection of males causes embryonic lethality unless they mate with similarly infected females, creating a relative reproductive advantage for infected females. A set of related Wolbachia bicistronic operons encodes the CI-inducing factors. The downstream gene encodes a deubiquitylase or nuclease and is responsible for CI induction by males, while the upstream product when expressed in females binds its sperm-introduced cognate partner and rescues viability. Both toxin-antidote and host-modification mechanisms have been proposed to explain CI. Interestingly, male killing by either Spiroplasma or Wolbachia endosymbionts involves deubiquitylases as well. Interference with the host ubiquitin system may therefore be a common theme among endosymbiont-mediated reproductive alterations.
Wolbachia-based strategies for control of agricultural pests
25104J. T. Gong, T. P. Li, M. K. Wang and X. Y. Hong, Curr Opin Insect Sci, 101039:10.1016/j.cois.2023.101039. 2023-04-25 10:26:15.
Wolbachia-based incompatible insect technique (IIT) and pathogen blocking technique (PBT) have been shown to be effective at protecting humans from mosquito-borne diseases in the past decades. Population suppression based on IIT and population replacement based on PBT have become major field application strategies that have continuously been improved by the translational research on Wolbachia-transinfected mosquitoes. Similarly, Wolbachia-based approaches have been proposed for the protection of plants from agricultural pests and their associated diseases. However, a bottleneck in Wolbachia-based strategies for the control of agricultural pests is the need for methods to establish Wolbachia-transinfected insect lines. As a first step in this direction, we compare field control strategies for mosquitos with the potential strategies for agricultural pests based on Wolbachia. Our results show that there is a critical need for establishing productive insect lines and accumulating field test data.
One strain may hide another: Cryptic male-killing Wolbachia
24924E. A. Hornett and G. D. D. Hurst, PLOS Biology, 21:e3002076. 2023-03-30 13:44:51.
While heritable symbionts are common in insects, strains that act as male-killers are considered rare. This Primer explores a new study in PLOS Biology which reveals a novel male-killer hidden by co-infection and host resistance, highlighting the complexity of host-microbial interactions in natural systems.
The Wolbachia WalE1 effector alters Drosophila endocytosis
24855M. Martin and I. L. G. Newton, bioRxiv, 2023-03-27 15:30:37.
The most common intracellular bacterial infection is Wolbachia pipientis , a microbe that manipulates host reproduction and is used in control of insect vectors. Phenotypes induced by Wolbachia have been studied for decades and range from sperm-egg incompatibility to male killing. How Wolbachia alters host biology is less well understood. Previously, we characterized the first Wolbachia effector â€" WalE1, which encodes a synuclein domain at the N terminus. Purified WalE1 sediments with and bundles actin and when heterologously expressed in flies, increases Wolbachia titer in the developing oocyte. In this work, we first identify the native expression WalE1 by Wolbachia infecting both fly cells and whole animals. WalE1 appears as aggregates, separate from Wolbachia cells. We next show that WalE1 co-immunoprecipitates with the host protein Past1 and that WalE1 manipulates host endocytosis. Yeast expressing WalE1 show deficiency in uptake of FM4-64 dye, and flies harboring mutations in Past1 or overexpressing WalE1 are sensitive to AgNO (3) , a hallmark of endocytosis defects. Finally, we also show that Past1 null flies harbor more Wolbachia overall and in late egg chambers. Our results identify interactions between a Wolbachia secreted effector and a host protein and point to yet another important host cell process impinged upon by Wolbachia .
Modelling Emergence of Wolbachia Toxin-Antidote Protein Functions with an Evolutionary Algorithm
24926J. Beckmann, J. Gillespie and D. Tauritz, bioRxiv, 2023-03-25 13:50:46.
Evolutionary algorithms (EAs) simulate Darwinian evolution and adeptly mimic natural evolution. Most EA applications in biology encode high levels of abstraction in top-down ecological population models. In contrast, our research merges protein alignment algorithms from bioinformatics into codon based EAs that simulate molecular protein string evolution from the bottom up. We apply our EA to reconcile a problem in the field of Wolbachia induced cytoplasmic incompatibility (CI). Wolbachia is a microbial endosymbiont that lives inside insect cells. CI is conditional insect sterility that operates as a toxin antidote (TA) system. Although, CI exhibits complex phenotypes not fully explained under a single discrete model. We instantiate in-silico genes that control CI, CI factors ( cifs ), as strings within the EA chromosome. We monitor the evolution of their enzymatic activity, binding, and cellular localization by applying selective pressure on their primary amino acid strings. Our model helps rationalize why two distinct mechanisms of CI induction might coexist in nature. We find that nuclear localization signals (NLS) and Type IV secretion system signals (T4SS) are of low complexity and evolve fast, whereas binding interactions have intermediate complexity, and enzymatic activity is the most complex. Our model predicts that as ancestral TA systems evolve into eukaryotic CI systems, the placement of NLS or T4SS signals can stochastically vary, imparting effects that might impact CI induction mechanics. Our model highlights how preconditions, genetic diversity, and sequence length can bias evolution of cifs towards one mechanism or another.
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.
Convergent Aedes and Drosophila CidB interactomes suggest cytoplasmic incompatibility targets are conserved
24875S. O. Oladipupo, J. D. Carroll and J. F. Beckmann, Insect Biochem Mol Biol, 103931. 2023-03-20 08:18:16.
Wolbachia-mediated cytoplasmic incompatibility (CI) is a conditional embryonic lethality induced when Wolbachia-modified sperm fertilizes an uninfected egg. The Wolbachia proteins, CidA and CidB control CI. CidA is a rescue factor that reverses lethality. CidA binds to CidB. CidB contains a deubiquitinating enzyme and induces CI. Precisely how CidB induces CI and what it targets are unknown. Likewise, how CidA prevents sterilization by CidB is not clear. To identify CidB substrates in mosquitos we conducted pull-down assays using recombinant CidA and CidB mixed with Aedes aegypti lysates to identify the protein interactomes of CidB and the CidB/CidA protein complex. Our data allow us to cross compare CidB interactomes across taxa for Aedes and Drosophila. Our data replicate several convergent interactions, suggesting that CI targets conserved substrates across insects. Our data support a hypothesis that CidA rescues CI by tethering CidB away from its substrates. Specifically, we identify ten convergent candidate substrates including P32 (protamine-histone exchange factor), karyopherin alpha, ubiquitin-conjugating enzyme, and bicoid stabilizing factor. Future analysis on how these candidates contribute to CI will clarify mechanisms.
Functional analysis of Wolbachia Cid effectors unravels cooperative interactions to target host chromatin during replication
24869K. Terretaz, B. Horard, M. Weill, B. Loppin and F. Landmann, PLOS Pathogens, 19:e1011211. 2023-03-16 08:02:08.
Wolbachia are common bacteria among terrestrial arthropods. These endosymbionts transmitted through the female germline manipulate their host reproduction through several mechanisms whose most prevalent form called Cytoplasmic Incompatibility -CI- is a conditional sterility syndrome eventually favoring the infected progeny. Upon fertilization, the sperm derived from an infected male is only compatible with an egg harboring a compatible Wolbachia strain, this sperm leading otherwise to embryonic death. The Wolbachia Cif factors CidA and CidB responsible for CI and its neutralization function as a Toxin-Antitoxin system in the mosquito host Culex pipiens. However, the mechanism of CidB toxicity and its neutralization by the CidA antitoxin remain unexplored. Using transfected insect cell lines to perform a structure-function analysis of these effectors, we show that both CidA and CidB are chromatin interactors and CidA anchors CidB to the chromatin in a cell-cycle dependent-manner. In absence of CidA, the CidB toxin localizes to its own chromatin microenvironment and acts by preventing S-phase completion, independently of its deubiquitylase -DUB- domain. Experiments with transgenic Drosophila show that CidB DUB domain is required together with CidA during spermatogenesis to stabilize the CidA-CidB complex. Our study defines CidB functional regions and paves the way to elucidate the mechanism of its toxicity.
NDUFA8 potentially rescues Wolbachia-induced cytoplasmic incompatibility in Laodelphax striatellus
24743J. Chen, M. K. Wang, Q. X. Xie, X. L. Bing, T. P. Li and X. Y. Hong, Insect Sci, 2023-02-06 18:20:35.
The endosymbiont Wolbachia manipulates host reproduction by several strategies, one of the most important of which is cytoplasmic incompatibility (CI). CI can be rescued when Wolbachia-infected (WI) males mate with females infected with the same Wolbachia strain. However, the potential rescue mechanism of CI in the small brown planthopper Laodelphax striatellus is unclear. In this study, comparative transcriptome analysis was applied to explore the effect of Wolbachia on L. striatellus eggs. A total of 1387 differentially expressed genes were identified. RNAi of seven Wolbachia-upregulated key planthopper genes reduced egg reproduction, suggesting that Wolbachia might improve fecundity in L. striatellus by affecting these seven genes. Suppressing the expression of another upregulated gene, NDUFA8 (encoding NADH dehydrogenase [ubiquinone] 1 alpha subcomplex subunit 8-like) by RNAi significantly increased the mortality of early embryos without affecting the number of deposited eggs. Wolbachia infection upregulated the mRNA level of NDUFA8, and dsNDUFA8 treatment of WI females re-created CI-like symptoms, suggesting that NDUFA8 is associated with the rescue phenotype. Because all L. striatellus populations worldwide are infected with Wolbachia, NDUFA8 is a potential pest control target. This article is protected by copyright. All rights reserved.
Pangenomic analysis of Wolbachia provides insight into the evolution of host adaptation and cytoplasmic incompatibility factor genes
24785B. Liu, Y. S. Ren, C. Y. Su, Y. Abe and D. H. Zhu, Frontiers in Microbiology, 14:1084839. 2023-02-03 09:44:36.
INTRODUCTION: The genus Wolbachia provides a typical example of intracellular bacteria that infect the germline of arthropods and filarial nematodes worldwide. Their importance as biological regulators of invertebrates, so it is particularly important to study the evolution, divergence and host adaptation of these bacteria at the genome-wide level. METHODS: Here, we used publicly available Wolbachia genomes to reconstruct their evolutionary history and explore their adaptation under host selection. RESULTS: Our findings indicate that segmental and single-gene duplications, such as DNA methylase, bZIP transcription factor, heat shock protein 90, in single monophyletic Wolbachia lineages (including supergroups A and B) may be responsible for improving the ability to adapt to a broad host range in arthropod-infecting strains. In contrast to A strains, high genetic diversity and rapidly evolving gene families occur in B strains, which may promote the ability of supergroup B strains to adapt to new hosts and their large-scale spreading. In addition, we hypothesize that there might have been two independent horizontal transfer events of cif genes in two sublineages of supergroup A strains. Interestingly, during the independent evolution of supergroup A and B strains, the rapid evolution of cif genes in supergroup B strains resulted in the loss of their functional domain, reflected in a possible decrease in the proportion of induced cytoplasmic incompatibility (CI) strains. DISCUSSION: This present study highlights for reconstructing of evolutionary history, addressing host adaptation-related evolution and exploring the origin and divergence of CI genes in each Wolbachia supergroup. Our results thus not only provide a basis for further exploring the evolutionary history of Wolbachia adaptation under host selection but also reveal a new research direction for studying the molecular regulation of Wolbachia- induced cytoplasmic incompatibility.
Single-cell transcriptome sequencing reveals Wolbachia-mediated modification in early stages of Drosophila spermatogenesis
24466W. Dou, B. Sun, Y. Miao, D. Huang and J. Xiao, Proceedings of the Royal Society B: Biological Sciences, 290:20221963. 2023-01-11 08:36:44.
Wolbachia are the most widely distributed intracellular bacteria, and their most common effect on host phenotype is cytoplasmic incompatibility (CI). A variety of models have been proposed to decipher the molecular mechanism of CI, among which the host modification (HM) model predicts that Wolbachia effectors play an important role in sperm modification. However, owing to the complexity of spermatogenesis and testicular cell-type heterogeneity, whether Wolbachia have different effects on cells at different stages of spermatogenesis or whether these effects are linked with CI remains unknown. Therefore, we used single-cell RNA sequencing to analyse gene expression profiles in adult male Drosophila testes that were infected or uninfected by Wolbachia. We found that Wolbachia significantly affected the proportion of different types of germ cells and affected multiple metabolic pathways in germ cells. Most importantly, Wolbachia had the greatest impact on germline stem cells, resulting in dysregulated expression of genes related to DNA compaction, and Wolbachia infection also influenced the histone-to-protamine transition in the late stage of sperm development. These results support the HM model and suggest that future studies on Wolbachia-induced CI should focus on cells in the early stages of spermatogenesis.
Wolbachia endosymbionts manipulate GSC self-renewal and differentiation to enhance host fertility
24270S. L. Russell, J. R. Castillo and W. T. Sullivan, bioRxiv, 2022.12.15.520626. 2022-12-17 14:03:40.
The alphaproteobacterium Wolbachia pipientis infects thousands of arthropod and nematode species worldwide, making it a key target for host biological control. Wolbachia-driven host reproductive manipulations, such as cytoplasmic incompatibility (CI), are often credited for catapulting these intracellular bacteria to high frequencies in their host populations. Positive, perhaps mutualistic, reproductive manipulations may also increase infection frequencies, but they are not well understood on molecular, cellular, or organismal levels. Previous studies demonstrated that Wolbachia is capable of partially rescuing sex-lethal, a gene required for germline stem cell (GSC) self renewal and bag-of-marbles, a gene required for GSC differentiation. Here, we identify molecular and cellular mechanisms by which Wolbachia is able to influence molecularly distinct processes of GSC self renewal and differentiation through our discovery that the wMel strain rescues meiotic-P26 mutants. Mei-P26 is an essential translational regulator and is required for both GSC self-renewal and differentiation. We demonstrate that wMel rescues the fertility of flies lacking adequate mei-P26 dosage and function, and is sufficient to sustain infertile homozygous mei-P26 hypomorphic stocks indefinitely. Cytology revealed that wMel infection mitigates the impact of mei-P26 loss on both germline stem cell maintenance and cyst differentiation through restoring proper pMad, Bam, Sxl, and Orb expression. Rescue in males, amplification of dominant negative effects, and multiallelic rescue suggest that wMel either directly or indirectly replaces Mei-P26 function. Even in wild-type individuals, wMel infection elevates lifetime egg lay and hatch rates. Over time, the beneficial fertility reinforcement mechanisms described here may promote the emergence of mutualism and the breakdown of CI.HighlightsThe wMel strain of Wolbachia restores fertility in females and males deficient for the essential translational regulator meiotic-P26Mei-P26’s germline maintenance and oocyte cyst differentiation functions are genetically rescued by wMel infectionPerturbed pMad, Sxl, Bam, and Orb expression are mitigated by wMel infectionwMel infection elevates lifetime egg lay and hatch rates in wild-type fliesCompeting Interest StatementThe authors have declared no competing interest.
Independent evaluation of Wolbachia infected male mosquito releases for control of Aedes aegypti in Harris County, Texas, using a Bayesian abundance estimator
23969S. Lozano, K. Pritts, D. Duguma, C. Fredregill and R. Connelly, PLOS Neglected Tropical Diseases, 16:e0010907. 2022-11-14 12:37:36.
Among disease vectors, Aedes aegypti (L.) (Diptera: Culicidae) is one of the most insidious species in the world. The disease burden created by this species has dramatically increased in the past 50 years, and during this time countries have relied on pesticides for control and prevention of viruses borne by Ae. aegypti. The small number of available insecticides with different modes of action had led to increases in insecticide resistance, thus, strategies, like the “Incompatible Insect Technique” using Wolbachia’s cytoplasmic incompatibility are desirable. We evaluated the effect of releases of Wolbachia infected Ae. aegypti males on populations of wild Ae. aegypti in the metropolitan area of Houston, TX. Releases were conducted by the company MosquitoMate, Inc. To estimate mosquito population reduction, we used a mosquito abundance Bayesian hierarchical estimator that accounted for inefficient trapping. MosquitoMate previously reported a reduction of 78% for an intervention conducted in Miami, FL. In this experiment we found a reduction of 93% with 95% credibility intervals of 86% and 96% after six weeks of continual releases. A similar result was reported by Verily Life Sciences, 96% [94%, 97%], in releases made in Fresno, CA.
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.
GeneConvene Global Collaborative Webinar Series | Wolbachia Biology, Mechanisms and Applications 2022
23723David O'Brochta, GeneConvene Global Collaborative, 2022-10-15 06:57:26.
Intracellular and extracellular symbiotic/commensal bacteria have enormous potential when manipulated and deployed appropriately to serve as agents of control of insects and the pathogens they transmit. Wolbachia, an intracellular bacteria, is a well studied system and one that is used increasingly to control insect populations via cytoplasmic incompatibility and to alter the vectoral capacity of insect via pathogen inhibition. This webinar series will survey the applications of Wolbachia as a mosquito/dengue control intervention and the underlying biology that conditions its effectiveness.
Wolbachia-Virus interactions and arbovirus control through population replacement in mosquitoes
23675T. H. Ant, M. V. Mancini, C. J. McNamara, S. M. Rainey and S. P. Sinkins, Pathogens and Global Health, 2022-10-07 08:27:10.
Following transfer into the primary arbovirus vector Aedes aegypti, several strains of the intracellular bacterium Wolbachia have been shown to inhibit the transmission of dengue, Zika, and chikungunya viruses, important human pathogens that cause significant morbidity and mortality worldwide. In addition to pathogen inhibition, many Wolbachia strains manipulate host reproduction, resulting in an invasive capacity of the bacterium in insect populations. This has led to the deployment of Wolbachia as a dengue control tool, and trials have reported significant reductions in transmission in release areas. Here, we discuss the possible mechanisms of Wolbachia-virus inhibition and the implications for long-term success of dengue control. We also consider the evidence presented in several reports that Wolbachia may cause an enhancement of replication of certain viruses under particular conditions, and conclude that these should not cause any concerns with respect to the application of Wolbachia to arbovirus control.
Estimating the effect of the wMel release programme on the incidence of dengue and chikungunya in Rio de Janeiro, Brazil: a spatiotemporal modelling study
23608G. Ribeiro dos Santos, B. Durovni, V. Saraceni, T. I. Souza Riback, S. B. Pinto, K. L. Anders, et al., The Lancet Infectious Diseases, 2022-09-28 09:07:02.
Summary Background Introgression of genetic material from species of the insect bacteria Wolbachia into populations of Aedes aegypti mosquitoes has been shown in randomised and non-randomised trials to reduce the incidence of dengue; however, evidence for the real-world effectiveness of large-scale deployments of Wolbachia-infected mosquitoes for arboviral disease control in endemic settings is still scarce. A large Wolbachia (wMel strain) release programme was implemented in 2017 in Rio de Janeiro, Brazil. We aimed to assess the effect of this programme on the incidence of dengue and chikungunya in the city. Methods 67 million wMel-infected mosquitoes were released across 28 489 locations over an area of 86·8 km2 in Rio de Janeiro between Aug 29, 2017 and Dec 27, 2019. Following releases, mosquitoes were trapped and the presence of wMel was recorded. In this spatiotemporal modelling study, we assessed the effect of the release programme on the incidence of dengue and chikungunya. We used spatiotemporally explicit mathematical models applied to geocoded dengue cases (N=283 270) from 2010 to 2019 and chikungunya cases (N=57 705) from 2016 to 2019. Findings On average, 32% of mosquitoes collected from the release zones between 1 month and 29 months after the initial release tested positive for wMel. Reduced wMel introgression occurred in locations and seasonal periods in which cases of dengue and chikungunya were historically high, with a decrease to 25% of mosquitoes testing positive for wMel during months in which disease incidence was at its highest. Despite incomplete introgression, we found that the releases were associated with a 38% (95% CI 32–44) reduction in the incidence of dengue and a 10% (4–16) reduction in the incidence of chikungunya. Interpretation Stable establishment of wMel in the geographically diverse, urban setting of Rio de Janeiro seems to be more complicated than has been observed elsewhere. However, even intermediate levels of wMel seem to reduce the incidence of disease caused by two arboviruses. These findings will help to guide future release programmes. Funding Bill & Melinda Gates Foundation and the European Research Council.
Wolbachia action in the sperm produces developmentally deferred chromosome segregation defects during the Drosophila mid-blastula transition
23615B. Warecki, S. W. A. Titen, M. S. Alam, G. Vega, N. Lemseffer, K. Hug, et al., eLife, 11:e81292. 2022-09-23 09:36:01.
Wolbachia, a vertically transmitted endosymbiont infecting many insects, spreads rapidly through uninfected populations by a mechanism known as cytoplasmic incompatibility (CI). In CI, a paternally delivered modification of the sperm leads to chromatin defects and lethality during and after the first mitosis of embryonic development in multiple species. However, whether CI-induced defects in later stage embryos are a consequence of the first division errors or caused by independent defects remains unresolved. To address this question, we focused on ~1/3 of embryos from CI crosses in Drosophila simulans that develop apparently normally through the first and subsequent pre-blastoderm divisions before exhibiting mitotic errors during the mid-blastula transition and gastrulation. We performed single embryo PCR and whole genome sequencing to find a large percentage of these developed CI-derived embryos bypass the first division defect. Using fluorescence in situ hybridization, we find increased chromosome segregation errors in gastrulating CI-derived embryos that had avoided the first division defect. Thus, Wolbachia action in the sperm induces developmentally deferred defects that are not a consequence of the first division errors. Like the immediate defect, the delayed defect is rescued through crosses to infected females. These studies inform current models on the molecular and cellular basis of CI.
Wolbachia strain wAlbB remains stable in Aedes aegypti over 15 years but exhibits genetic background-dependent variation in virus blocking
23646X. Liang, C. H. Tan, Q. Sun, M. Zhang, P. J. Wong, M. I. Li, et al., PNAS Nexus, 2022-09-22 08:20:54.
The ability of the maternally transmitted endosymbiotic bacterium Wolbachia to induce cytoplasmic incompatibility (CI) and virus blocking makes it a promising weapon for combatting mosquito-borne diseases through either suppression or replacement of wild-type populations. Recent field trials show that both approaches significantly reduce the incidence of dengue fever in humans. However, new questions emerge about how Wolbachia-mosquito associations will co-evolve over time and whether Wolbachia-mediated virus blocking will be affected by the genetic diversity of mosquitoes and arboviruses in the real world. Here, we have compared the Wolbachia density and CI expression of two wAlbB-infected Aedes aegypti lines transinfected 15 years apart. We have also assessed wAlbB-mediated virus blocking against dengue (DENV), Zika (ZIKV), and Chikungunya (CHIKV) viruses and examined whether host genetic backgrounds modulate viral blocking effects by comparing ZIKV infection in mosquitoes with a Mexican genetic background to those with a Singaporean background. Our results show that over 15 years, wAlbB maintained the capacity to form a stable association with Ae. aegypti in terms of both density and CI expression. There were variations in wAlbB-induced virus blocking against CHIKV, DENV, and ZIKV, and higher inhibitory effects on ZIKV in mosquitoes on the Singaporean genetic background than on the Mexican background. These results provide important information concerning the robustness and long-term stability of Wolbachia as a biocontrol agent for arbovirus disease control.
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.
Comparative Ubiquitome Analysis Reveals Deubiquitinating Effects Induced by Wolbachia Infection in Drosophila melanogaster
23489Q. Zong, B. Mao, H. B. Zhang, B. Wang, W. J. Yu, Z. W. Wang and Y. F. Wang, International Journal Molecular Science, 23. 2022-08-21 07:14:27.
The endosymbiotic Wolbachia bacteria frequently cause cytoplasmic incompatibility (CI) in their insect hosts, where Wolbachia-infected males cross with uninfected females, leading to no or fewer progenies, indicating a paternal modification by Wolbachia. Recent studies have identified a Wolbachia protein, CidB, containing a DUB (deubiquitylating enzyme) domain, which can be loaded into host sperm nuclei and involved in CI, though the DUB activity is not necessary for CI in Drosophila melanogaster. To investigate whether and how Wolbachia affect protein ubiquitination in testes of male hosts and are thus involved in male fertility, we compared the protein and ubiquitinated protein expressions in D. melanogaster testes with and without Wolbachia. A total of 643 differentially expressed proteins (DEPs) and 309 differentially expressed ubiquitinated proteins (DEUPs) were identified to have at least a 1.5-fold change with a p-value of <0.05. Many DEPs were enriched in metabolic pathway, ribosome, RNA transport, and post-translational protein modification pathways. Many DEUPs were involved in metabolism, ribosome, and proteasome pathways. Notably, 98.1% DEUPs were downregulated in the presence of Wolbachia. Four genes coding for DEUPs in ubiquitin proteasome pathways were knocked down, respectively, in Wolbachia-free fly testes. Among them, Rpn6 and Rpn7 knockdown caused male sterility, with no mature sperm in seminal vesicles. These results reveal deubiquitylating effects induced by Wolbachia infection, suggesting that Wolbachia can widely deubiquitinate proteins that have crucial functions in male fertility of their hosts, but are not involved in CI. Our data provide new insights into the regulatory mechanisms of endosymbiont/host interactions and male fertility.
Harnessing Wolbachia cytoplasmic incompatibility alleles for confined gene drive: a modeling study
23424J. Li and J. Champer, bioRxiv, 2022.08.09.503337. 2022-08-11 07:25:47.
By using both mathematical and simulation models, we found that a drive containing CifA and CifB together create a confined drive with a moderate to high introduction threshold. When introduced separately, they act as a self-limiting drive. We observed that the performance of these drives is substantially influenced by various ecological parameters and drive characteristics. Extending our models to continuous space, we found that the drive individual release distribution has a critical impact on drive persistence.
Developing Wolbachia-based disease interventions for an extreme environment
23309P. A. Ross, S. Elfekih, S. Collier, M. J. Klein, S. S. Lee, M. Dunn, S. Jackson, Y. Zhang, J. K. Axford, X. Gu, M. S. Nasar, P. N. Paradkar, E. A. Taoufik, F. M. Jiggins, A. M. Almalik, M. B. Al-Fageeh and A. A. Hoffmann, bioRxiv, 2022.07.26.501527. 2022-07-27 07:59:12.
Aedes aegypti mosquitoes carrying self-spreading, virus-blocking Wolbachia bacteria are being deployed to suppress dengue transmission. However, there are challenges in applying this technology in extreme environments. We introduced two Wolbachia strains into Ae. aegypti from Saudi Arabia for a release program in the hot coastal city of Jeddah. Wolbachia reduced infection and dissemination of dengue virus (DENV2) in Saudi Arabian mosquitoes and showed complete maternal transmission and cytoplasmic incompatibility. Wolbachia reduced mosquito heat tolerance and egg viability, with the Wolbachia strains showing differential thermal stability. Wolbachia effects were similar across mosquito genetic backgrounds but we found evidence of local adaptation, with Saudi Arabian mosquitoes having lower egg viability but higher adult desiccation tolerance than Australian mosquitoes. Genetic background effects will influence Wolbachia invasion dynamics, reinforcing the need to use local genotypes for mosquito release programs, particularly in extreme environments like Jeddah. Our comprehensive characterization of Wolbachia strains provides a foundation for Wolbachia-based disease interventions in harsh climates.Competing Interest StatementThe authors have declared no competing interest.
Attempts to use breeding approaches in Aedes aegypti to create lines with distinct and stable relative Wolbachia densities
23279A. J. Mejia, L. Jimenez, H. L. C. Dutra, R. Perera and E. A. McGraw, Heredity, 2022-07-22 10:12:25.
Wolbachia is an insect endosymbiont being used for biological control in the mosquito Aedes aegypti because it causes cytoplasmic incompatibility (CI) and limits viral replication of dengue, chikungunya, and Zika viruses. While the genetic mechanism of pathogen blocking (PB) is not fully understood, the strength of both CI and PB are positively correlated with Wolbachia densities in the host. Wolbachia densities are determined by a combination of Wolbachia strain and insect genotype, as well as interactions with the environment. We employed both artificial selection and inbreeding with the goal of creating lines of Ae. aegypti with heritable and distinct Wolbachia densities so that we might better dissect the mechanism underlying PB. We were unable to shift the mean relative Wolbachia density in Ae. aegypti lines by either strategy, with relative densities instead tending to cycle over a narrow range. In lieu of this, we used Wolbachia densities in mosquito legs as predictors of relative densities in the remaining individual’s carcass. Because we worked with outbred mosquitoes, our findings indicate either a lack of genetic variation in the mosquito for controlling relative density, natural selection against extreme densities, or a predominance of environmental factors affecting densities. Our study reveals that there are moderating forces acting on relative Wolbachia densities that may help to stabilize density phenotypes post field release. We also show a means to accurately bin vector carcasses into high and low categories for non-DNA omics-based studies of Wolbachia-mediated traits.
cifB-transcript levels largely explain cytoplasmic incompatibility variation across divergent Wolbachia
23074J. D. Shropshire, E. Hamant, W. R. Conner and B. S. Cooper, PNAS Nexus, 2022-06-28 13:29:28.
Divergent hosts often associate with intracellular microbes that influence their fitness. Maternally transmitted Wolbachia bacteria are the most common of these endosymbionts due largely to cytoplasmic incompatibility (CI) that kills uninfected embryos fertilized by Wolbachia-infected males. Closely related infections in females rescue CI, providing a relative fitness advantage that drives Wolbachia to high frequencies. One prophage-associated gene (cifA) governs rescue and two contribute to CI (cifA and cifB), but CI strength ranges from very strong to very weak for unknown reasons. Here, we investigate CI-strength variation and its mechanistic underpinnings in a phylogenetic context across 20 million years (MY) of Wolbachia evolution in Drosophila hosts diverged up to 50 MY. These Wolbachia encode diverse Cif proteins (100–7.4% pairwise similarity), and AlphaFold structural analyses suggest that CifB sequence similarities do not predict structural similarities. We demonstrate that cifB-transcript levels in testes explain CI strength across all but two focal systems. Despite phylogenetic discordance among cifs and the bulk of the Wolbachia genome, closely related Wolbachia tend to cause similar CI strengths and transcribe cifB at similar levels. This indicates that other non-cif regions of the Wolbachia genome modulate cif-transcript levels. CI strength also increases with the length of the host's larval life stage, presumably due to prolonged cif action. Our findings reveal that cifB-transcript levels largely explain CI strength while highlighting other covariates. Elucidating CI's mechanism contributes to our understanding of Wolbachia spread in natural systems and the efficacy of CI-based biocontrol of arboviruses and agricultural pests globally.
Sensitivity of wMel and wAlbB Wolbachia infections in Aedes aegypti Puducherry (Indian) strains to heat stress during larval development
23056K. Gunasekaran, C. Sadanandane, D. Panneer, A. Kumar, M. Rahi, S. Dinesh, B. Vijayakumar, M. Krishnaraja, S. K. Subbarao and P. Jambulingam, Parasites and Vectors, 15:221. 2022-06-21 15:19:54.
BACKGROUND: ICMR-Vector Control Research Centre, Puducherry, India, developed two colonies of Aedes aegypti infected with wMel and wAlbB Wolbacia strains called Ae. aegypti (Pud) lines for dengue control. The sensitivity of wMel and wAlbB strains in Ae. aegypti (Pud) lines to heat stress was studied. METHODS: wMel and wAlbB infected and uninfected Ae. aegypti larvae (first to fourth instars) were reared in the laboratory to adults at 26 °C, 30 °C, 36 °C and 40 °C constant temperatures and also 26-30 °C, 26-36 °C and 26-40 °C diurnal cyclic temperatures. The adults were tested for Wolbachia infection. Experiments were also carried out rearing the larvae under simulated field conditions in summer (April and June) under sunlight using fully open and half open bowls and also under sunlight and natural shade. RESULTS: At 36 °C and 40 °C constant temperatures, complete larval mortality was observed. At 30 °C and 26 °C, no larval mortality occurred, but Wolbachia density was relatively low in wMel infected males compared to control (maintained at 26 ± 1 °C). At diurnal cyclic temperature of 26-40 °C, Wolbachia density was reduced in males of both the (Pud) lines, but not in females. At 26-36 °C, reduction in Wolbachia density was observed in wMel males but not in wAlbB males. At 26-30 °C, no significant reduction in Wolbachia density was observed with wMel and wAlbB strains. In simulated field conditions (April), under sunlight, the daytime water temperature reached a maximum of 35.7 °C in both full and half open bowls. No larval mortality occurred. Wolbachia frequency and density was reduced in wMel-infected Ae. aegypti (Pud) males from both type of bowls and in females from full open bowls, and in wAlbB males from half open bowls. In June, rearing of larvae under sunlight, the first-instar larvae experienced a maximum daytime water temperature of > 38 °C that caused complete mortality. No larval mortality was observed in bowls kept under shade (< 32 °C). CONCLUSIONS: Exposure of larvae to higher rearing temperatures in the laboratory and simulated-field conditions reduced the densities of wMel and wAlbB strains particularly in males, but the impact was more pronounced for wMel strain. The actual effect of heat stress on the stability of these two Wolbachia strains needs to be tested under natural field conditions.
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.
Mosquito control to save Hawaiian honeycreepers does not involve GMOs
22859Department of Land and Natural Resources, Hawaii Department of Land and Natural Resources, 2022-06-08 08:42:00.
Despite misinformation circulating on social media, the importation of “incompatible-male” mosquitoes to control populations of wild mosquitoes and to save four native bird species from extinction, does not involve the use of any genetically modified organisms (GMOs) or genetically engineered (GE) organisms. On Thursday, the Plant and Animal Advisory Committee of the Dept. of Agriculture will consider listing three species of mosquitoes on its Restricted Species List A. The listing would allow the importation of three species of mosquitoes, all of which are already present in Hawai‘i. One of these, the Southern House Mosquito (Culex quinquefasciatus) isresponsible for sharp declines in the populations of many honeycreeper species on Kaua‘i, Maui, and Hawai‘i Island. The other two species – Yellow Fever Mosquito (Aedes aegypti) and Asian Tiger Mosquito (Aedes albopictus) – transmit human diseases.
Establishment of Wolbachia infection in Aedes aegypti from Pakistan via embryonic microinjection and semi-field evaluation of general fitness of resultant mosquito population
22856M. S. Sarwar, N. Jahan, A. Ali, H. K. Yousaf and I. Munzoor, Parasites and Vectors, 15:191. 2022-06-06 08:36:19.
BACKGROUND: Dengue is a mosquito-borne viral disease that is mainly spread by Aedes aegypti. It is prevalent on five continents, predominantly in tropical and sub-tropical zones across the world. Wolbachia bacteria have been extensively used in vector control strategies worldwide. The focus of the current study was to obtain a natural population of Ae. aegypti harbouring Wolbachia and to determine the impact of this bacteria on the new host in a semi-field environment. METHODS: Wolbachia-infected Aedes albopictus was collected from the city of Lahore, Punjab, Pakistan, and Wolbachia were successfully introduced into laboratory-reared Ae. aegypti via embryonic microinjection. The stable vertical transmission of wAlbB in the host population was observed for eight generations, and the impact of Wolbachia on the general fitness of the host was evaluated in semi-field conditions. RESULTS: In the laboratory and semi-field experiments, wAlbB Wolbachia presented a strong cytoplasmic incompatibility (CI) effect, evidenced as zero egg hatching, in crosses between Wolbachia-infected males and wild (uninfected) females of Ae. aegypti. Wolbachia infection had no noticeable impact on the general fitness (P > 0.05), fecundity, body size (females and males) and mating competitiveness of the new host, Ae. aegypti. However, there was a significant decrease in female fertility (egg hatch) (P < 0.001). In addition, under starvation conditions, there was a remarkable decrease (P < 0.0001) in the life span of Wolbachia-infected females compared to uninfected females (4 vs. > 5 days, respectively). CONCLUSIONS: Wolbachia strain wAlbB has a great potential to control the dengue vector in Ae. aegypti populations by producing 100% CI with a limited burden on its host in natural field conditions. This strain can be used as a biological tool against vector-borne diseases.
Perplexing dynamics of Wolbachia proteins for cytoplasmic incompatibility
22582T. Harumoto and T. Fukatsu, PLOS Biology, 20:e3001644. 2022-05-25 09:23:59.
The mechanism of symbiont-induced cytoplasmic incompatibility has been a long-lasting mystery. This Primer explores a new study on Wolbachia’s Cif proteins in PLOS Biology that provides supportive evidence for the “Host-Modification Model,” although the alternative “Toxin-Antidote Model” is still in the running.
The Cif proteins from Wolbachia prophage WO modify sperm genome integrity to establish cytoplasmic incompatibility
22570R. Kaur, B. A. Leigh, I. T. Ritchie and S. R. Bordenstein, PLOS Biology, 20:e3001584. 2022-05-24 08:55:13.
In Drosophila melanogaster, germline expression of the Wolbachia proteins CifA and CifB causes cytoplasmic incompatibility, but how this impairs male reproduction remains unclear. This study shows how Cif proteins encoded by the endosymbiont prophage invade the fly’s gametic nuclei and modify the histone-protamine transition of spermatogenesis.
Strategies to Mitigate Establishment under the Wolbachia Incompatible Insect Technique
23104S. Soh, S. H. Ho, J. Ong, A. Seah, B. S. Dickens, K. W. Tan, J. R. Koo, A. R. Cook, S. Sim, C. H. Tan, L. C. Ng and J. T. Lim, Viruses, 14. 2022-05-24 08:10:53.
The Incompatible Insect Technique (IIT) strategy involves the release of male mosquitoes infected with the bacterium Wolbachia. Regular releases of male Wolbachia-infected mosquitoes can lead to the suppression of mosquito populations, thereby reducing the risk of transmission of vector-borne diseases such as dengue. However, due to imperfect sex-sorting under IIT, fertile Wolbachia-infected female mosquitoes may potentially be unintentionally released into the environment, which may result in replacement and failure to suppress the mosquito populations. As such, mitigating Wolbachia establishment requires a combination of IIT with other strategies. We introduced a simple compartmental model to simulate ex-ante mosquito population dynamics subjected to a Wolbachia-IIT programme. In silico, we explored the risk of replacement, and strategies that could mitigate the establishment of the released Wolbachia strain in the mosquito population. Our results suggest that mitigation may be achieved through the application of a sterile insect technique. Our simulations indicate that these interventions do not override the intended wild type suppression of the IIT approach. These findings will inform policy makers of possible ways to mitigate the potential establishment of Wolbachia using the IIT population control strategy.
Cytoplasmic incompatibility: A Wolbachia toxin–antidote mechanism comes into view
21546M. Hochstrasser, Current Biology, 32:R287-R289. 2022-03-28 11:56:01.
The Wolbachia cidA and cidB genes promote bacterial endosymbiont inheritance through the host female germline. CidB is now shown to load into maturing sperm nuclei. Following fertilization, it disrupts paternal chromosome condensation, triggering embryonic arrest if not countered by CidA in Wolbachia-infected eggs.
Uniqueness and stability of periodic solutions for an interactive wild and Wolbachia-infected male mosquito model
20438R. Yan and Q. Sun, Journal of Biological Dynamics, 2022-02-15 09:03:48.
We investigate a mosquito population suppression model, which includes the release of Wolbachia-infected males causing incomplete cytoplasmic incompatibility (CI). The model consists of two sub-equations by considering the density-dependent birth rate of wild mosquitoes. By assuming the release waiting period T is larger than the sexual lifespan T¯ of Wolbachia-infected males, we derive four thresholds: the CI intensity threshold sh∗, the release amount thresholds g∗ and c∗, and the waiting period threshold T∗. From a biological view, we assume sh > sh∗ throughout the paper. When g∗ < c < c∗, we prove the origin E0 is locally asymptotically stable iff T < T∗, and the model admits a unique T-periodic solution iff T ≥ T∗, which is globally asymptotically stable. When c ≥ c∗, we show the origin E0 is globally asymptotically stable iff T ≤ T∗, and the model has a unique T-periodic solution iff T > T∗, which is globally asymptotically stable. Our theoretical results are confirmed by numerical simulations.
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.
Endosymbionts moderate constrained sex allocation in a haplodiploid thrips species in a temperature-sensitive way
20394A. Katlav, D. T. Nguyen, J. L. Morrow, R. N. Spooner-Hart and M. Riegler, Heredity, 9. 2022-02-03 09:32:33.
Maternally inherited bacterial endosymbionts that affect host fitness are common in nature. Some endosymbionts colonise host populations by reproductive manipulations (such as cytoplasmic incompatibility; CI) that increase the reproductive fitness of infected over uninfected females. Theory predicts that CI-inducing endosymbionts in haplodiploid hosts may also influence sex allocation, including in compatible crosses, however, empirical evidence for this is scarce. We examined the role of two common CI-inducing endosymbionts, Cardinium and Wolbachia, in the sex allocation of Pezothrips kellyanus, a haplodiploid thrips species with a split sex ratio. In this species, irrespective of infection status, some mated females are constrained to produce extremely male-biased broods, whereas other females produce extremely female-biased broods. We analysed brood sex ratio of females mated with males of the same infection status at two temperatures. We found that at 20 degrees C the frequency of constrained sex allocation in coinfected pairs was reduced by 27% when compared to uninfected pairs. However, at 25 degrees C the constrained sex allocation frequency increased and became similar between coinfected and uninfected pairs, resulting in more male-biased population sex ratios at the higher temperature. This temperature-dependent pattern occurred without changes in endosymbiont densities and compatibility. Our findings indicate that endosymbionts affect sex ratios of haplodiploid hosts beyond the commonly recognised reproductive manipulations by causing female-biased sex allocation in a temperature-dependent fashion. This may contribute to a higher transmission efficiency of CI-inducing endosymbionts and is consistent with previous models that predict that CI by itself is less efficient in driving endosymbiont invasions in haplodiploid hosts.
Paternal transmission of the Wolbachia CidB toxin underlies cytoplasmic incompatibility
20434B. Horard, K. Terretaz, A. S. Gosselin-Grenet, H. Sobry, M. Sicard, F. Landmann and B. Loppin, Current Biology, 2022-02-01 08:53:36.
Wolbachia are widespread endosymbiotic bacteria that manipulate the reproduction of arthropods through a diversity of cellular mechanisms. In cytoplasmic incompatibility (CI), a sterility syndrome originally discovered in the mosquito Culex pipiens, uninfected eggs fertilized by sperm from infected males are selectively killed during embryo development following the abortive segregation of paternal chromosomes in the zygote. Despite the recent discovery of Wolbachia CI factor (cif) genes, the mechanism by which they control the fate of paternal chromosomes at fertilization remains unknown. Here, we have analyzed the cytological distribution and cellular impact of CidA and CidB, a pair of Cif proteins from the Culex-infecting Wolbachia strain wPip. We show that expression of CidB in Drosophila S2R+ cells induces apoptosis unless CidA is co-expressed and associated with its partner. In transgenic Drosophila testes, both effectors colocalize in germ cells until the histone-to-protamine transition in which only CidB is retained in maturing spermatid nuclei. We further show that CidB is similarly targeted to maturing sperm of naturally infected Culex mosquitoes. At fertilization, CidB associates with paternal DNA regions exhibiting DNA replication stress, as a likely cause of incomplete replication of paternal chromosomes at the onset of the first mitosis. Importantly, we demonstrate that inactivation of the deubiquitylase activity of CidB does not abolish its cell toxicity or its ability to induce CI in Drosophila. Our study thus demonstrates that CI functions as a transgenerational toxin-antidote system and suggests that CidB acts by poisoning paternal DNA replication in incompatible crosses.
Cytoplasmic incompatibility in hybrid zones: infection dynamics and resistance evolution
19742E. S. Røed and J. Engelstädter, Journal of Evolutionary Biology, 2021-12-25 13:16:15.
Cytoplasmic incompatibility is an endosymbiont-induced mating incompatibility common in arthropods. Unidirectional cytoplasmic incompatibility impairs crosses between infected males and uninfected females, whereas bidirectional cytoplasmic incompatibility occurs when two host lineages are infected with reciprocally in compatible endosymbionts. Bidirectional cytoplasmic incompatibility is unstable in unstructured populations, but may be stable in hybrid zones. Stable coexistence of incompatible host lineages should generate frequent incompatible crosses. Therefore, hosts are expected to be under selection to resist their endosymbionts. Here, we for mulate a mathematical model of hybrid zones where two bidirectionally incompatible host lineages meet. We expand this model to consider the invasion of a hypothetical resistance allele. To corroborate our mathematical predictions, we test each prediction with stochastic, individual-based simulations. Our models suggest that hybrid zones may sustain stable coinfections of bidirectionally incompatible endosymbiont strains. Over a range of conditions, host are under selection for resistance against cytoplasmic incompatibility. Under asymetric migration, a resistance allele can facilitate infection turnover and subsequently either persist or become lost. The predictions we present may inform our understanding of the cophylogenetic relationship between the endosym biont Wolbachia and its hosts.
From Wolbachia genomics to phenotype: molecular models of cytoplasmic incompatibility must account for the multiplicity of compatibility types
19733A. Namias, M. Sicard, M. Weill and S. Charlat, Current Opinion in Insect Science, 2021-12-23 13:03:28.
Wolbachia endosymbionts commonly induce cytoplasmic incompatibility, making infected males’ sperm lethal to the embryos unless these are rescued by the same bacterium, inherited from their mother. Causal genes were recently identified but two families of mechanistic models are still opposed. In the toxin–antidote model, interaction between the toxin and the antidote is required for rescuing the embryos. In host modification models, a host factor is misregulated in sperm and rescue occurs through compensation or withdrawal of this modification. While these models have been thoroughly discussed, the multiplicity of compatibility types, that is, the existence of many mutually incompatible strains, as seen in Culex mosquitoes, has not received sufficient attention. To explain such a fact, host modification models must posit that the same embryonic defects can be induced and rescued through a large variety of host targets. Conversely, the toxin–antidote model simply accommodates this pattern in a lock–key fashion, through variations in the toxin–antidote interaction sites.
Wolbachia Endosymbiont and Mosquito Vectors, with Emphasis on Lymphatic Filariasis Elimination
19965I. P. Sunish, Genetically Modified and other Innovative Vector Control Technologies, 2021-12-21 11:24:11.
Wolbachia are maternally inherited intracellular bacteria, known to alter early development and mitotic processes in their hosts. They are frequently observed as a reproductive parasite, capable of inducing feminization, parthenogenesis, male killing, or cytoplasmic incompatibility. A total of 18 clades of Wolbachia have been reported, almost exclusively in arthropods. Wolbachia-based strategies have been proposed for the control of disease vectors. Wolbachia-based population suppression and transmission blocking can work in species not commonly infected with Wolbachia in the wild. However, efficient maintenance and spread of Wolbachia infection into field populations is crucial to the success of this strategy. Property of cytoplasmic incompatibility (CI) can be used to reduce the density of mosquito field populations through inundative releases of incompatible males in order to sterilize females. In semi-field condition at La Reunion, the LR[wPip(Is)] males of Culex quinquefasciatus successfully competed with field males in mating with field females. Depletion of Wolbachia endobacteria by antibiotic therapy prevents larval moulting and kills adult filarial worms. This strategy could act as an adjunct to vector control and is being exploited for the elimination of lymphatic filariasis.
Combined sterile insect technique and incompatible insect technique: concept, study design, experience and lessons learned from a pilot suppression trial in Thailand
19367Kittayapong, P., AREA-WIDE INTEGRATED PEST MANAGEMENT: Development and Field Application, 2021-11-29 16:50:12.
Climate change, rapid global transport and land use change leading to urbanization and agricultural intensification have facilitated disease emergence in vulnerable regions like Southeast Asia, and also the global expansion of vectors and vector-borne diseases into other regions like the Americas and Europe. Important vector-borne diseases, i.e. dengue, chikungunya, yellow fever, and Zika are transmitted by the major mosquito vector species, Aedes aegypti (L.) and Aedes albopictus (Skuse). Management of Ae. aegypti populations in countries endemic to these diseases, especially in Southeast Asia, is not sufficiently effective, resulting in high morbidity and mortality in the region. Insecticide resistance has become an important issue, causing failure in insecticide-based vector control. Innovative or alternative tools/approaches are needed to effectively reduce mosquito vector populations and consequently reduce the diseases they transmit. A trial integrating the environment-friendly Sterile Insect Technique (SIT) and the insect incompatible technique (IIT) was successfully carried out on a small-scale in a semi-rural setting in Thailand. In this chapter, we report on the design and methodology, as well as the experience and lessons learned from the baseline preparation and implementation of the pilot trial.
Wolbachia cifB induces cytoplasmic incompatibility in the malaria mosquito vector
19439K. L. Adams, D. G. Abernathy, B. C. Willett, E. K. Selland, M. A. Itoe and F. Catteruccia, Nature Microbiology, 6:1575-1582. 2021-11-26 20:50:59.
Wolbachia, a maternally inherited intracellular bacterial species, can manipulate host insect reproduction by cytoplasmic incompatibility (CI), which results in embryo lethality in crosses between infected males and uninfected females. CI is encoded by two prophage genes, cifA and cifB. Wolbachia, coupled with the sterile insect technique, has been used in field trials to control populations of the dengue vector Aedes albopictus, but CI-inducing strains are not known to infect the malaria vector Anopheles gambiae. Here we show that cifA and cifB can induce conditional sterility in the malaria vector An. gambiae. We used transgenic expression of these Wolbachia-derived genes in the An. gambiae germline to show that cifB is sufficient to cause embryonic lethality and that cifB-induced sterility is rescued by cifA expression in females. When we co-expressed cifA and cifB in male mosquitoes, the CI phenotype was attenuated. In female mosquitoes, cifB impaired fertility, which was overcome by co-expression of cifA. Our findings pave the way towards using CI to control malaria mosquito vectors.
High Temperature Cycles Result in Maternal Transmission and Dengue Infection Differences Between Wolbachia Strains in Aedes aegypti
19221M. V. Mancini, T. H. Ant, C. S. Herd, J. Martinez, S. M. Murdochy, D. D. Gingell, E. Mararo, P. C. D. Johnson and S. P. Sinkins, mBio, e0025021. 2021-11-10 21:39:39.
Environmental factors play a crucial role in the population dynamics of arthropod endosymbionts, and therefore in the deployment of Wolbachia symbionts for the control of dengue arboviruses. The potential of Wolbachia to invade, persist, and block virus transmission depends in part on its intracellular density. Several recent studies have highlighted the importance of larval rearing temperature in modulating Wolbachia densities in adults, suggesting that elevated temperatures can severely impact some strains, while having little effect on others. The effect of a replicated tropical heat cycle on Wolbachia density and levels of virus blocking was assessed using Aedes aegypti lines carrying strains wMel and wAlbB, two Wolbachia strains currently used for dengue control. Impacts on intracellular density, maternal transmission fidelity, and dengue inhibition capacity were observed for wMel. In contrast, wAlbB-carrying Ae. aegypti maintained a relatively constant intracellular density at high temperatures and conserved its capacity to inhibit dengue. Following larval heat treatment, wMel showed a degree of density recovery in aging adults, although this was compromised by elevated air temperatures. IMPORTANCE In the past decades, dengue incidence has dramatically increased all over the world. An emerging dengue control strategy utilizes Aedes aegypti mosquitoes artificially transinfected with the bacterial symbiont Wolbachia, with the ultimate aim of replacing wild mosquito populations. However, the rearing temperature of mosquito larvae is known to impact on some Wolbachia strains. In this study, we compared the effects of a temperature cycle mimicking natural breeding sites in tropical climates on two Wolbachia strains, currently used for open field trials. When choosing the Wolbachia strain to be used in a dengue control program it is important to consider the effects of environmental temperatures on invasiveness and virus inhibition. These results underline the significance of understanding the impact of environmental factors on released mosquitoes, in order to ensure the most efficient strategy for dengue control.
Novel Symbiotic Genome-Scale Model Reveals Wolbachia’s Arboviral Pathogen Blocking Mechanism in Aedes aegypti
18936N. E. Jiménez, Z. P. Gerdtzen, Á. Olivera-Nappa, J. C. Salgado and C. Conca, mBio, e0156321. 2021-10-13 20:29:16.
Arboviral diseases such as Zika and Dengue have been on the rise mainly due to climate change, and the development of new treatments and strategies to limit their spreading is needed. The use of Wolbachia as an approach for disease control has motivated new research related to the characterization of the mechanisms that underlie its pathogen-blocking properties. In this work, we propose a new approach for studying the metabolic interactions between Aedes aegypti and Wolbachia using genome-scale models, finding that pathogen blocking is mainly influenced by competition for the resources required for Wolbachia and viral replication.
Structural and mechanistic insights into the complexes formed by Wolbachia cytoplasmic incompatibility factors
18908Y. Xiao, H. Chen, H. Wang, M. Zhang, X. Chen, J. M. Berk, L. Zhang, Y. Wei, W. Li, W. Cui, F. Wang, Q. Wang, C. Cui, T. Li, C. Chen, S. Ye, L. Zhang, X. Ji, J. Huang, W. Wang, Z. Wang, M. Hochstrasser and H. Yang, Proceedings of the National Academy of Sciences, 118. 2021-10-09 14:35:56.
Wolbachia bacteria, inherited through the female germ line, infect a large fraction of arthropod species. Many Wolbachia strains manipulate host reproduction, most commonly through cytoplasmic incompatibility (CI). CI, a conditional male sterility, results when Wolbachia-infected male insects mate with uninfected females; viability is restored if the female is similarly infected (called "rescue"). CI is used to help control mosquito-borne viruses such as dengue and Zika, but its mechanisms remain unknown. The coexpressed CI factors CifA and CifB form stable complexes in vitro, but the timing and function of this interaction in the insect are unresolved. CifA expression in the female germ line is sufficient for rescue. We report high-resolution structures of a CI-factor complex, CinA-CinB, which utilizes a unique binding mode between the CinA rescue factor and the CinB nuclease; the structures were validated by biochemical and yeast growth analyses. Importantly, transgenic expression in Drosophila of a nonbinding CinA mutant, designed based on the CinA-CinB structure, suggests CinA expressed in females must bind CinB imported by sperm in order to rescue embryonic viability. Binding between cognate factors is conserved in an enzymatically distinct CI system, CidA-CidB, suggesting universal features in Wolbachia CI induction and rescue.
A single mutation weakens symbiont-induced reproductive manipulation through reductions in deubiquitylation efficiency
18742J. F. Beckmann, K. Van Vaerenberghe, D. E. Akwa and B. S. Cooper, Proceedings of the National Academy of Sciences, 118:e2113271118. 2021-09-21 14:01:04.
We show that a single naturally observed mutation weakens CI by reducing deubiquitylation. These discoveries help elucidate the molecular basis of symbiont-induced reproductive manipulations.Animals interact with microbes that affect their performance and fitness, including endosymbionts that reside inside their cells. Maternally transmitted Wolbachia bacteria are the most common known endosymbionts, in large part because of their manipulation of host reproduction. For example, many Wolbachia cause cytoplasmic incompatibility (CI) that reduces host embryonic viability when Wolbachia-modified sperm fertilize uninfected eggs. Operons termed cifs control CI, and a single factor (cifA) rescues it, providing Wolbachia-infected females a fitness advantage. Despite CI’s prevalence in nature, theory indicates that natural selection does not act to maintain CI, which varies widely in strength. Here, we investigate the genetic and functional basis of CI-strength variation observed among sister Wolbachia that infect Drosophila melanogaster subgroup hosts. We cloned, Sanger sequenced, and expressed cif repertoires from weak CI–causing wYak in Drosophila yakuba, revealing mutations suspected to weaken CI relative to model wMel in D. melanogaster. A single valine-to-leucine mutation within the deubiquitylating (DUB) domain of the wYak cifB homolog (cidB) ablates a CI-like phenotype in yeast. The same mutation reduces both DUB efficiency in vitro and transgenic CI strength in the fly, each by about twofold. Our results map hypomorphic transgenic CI to reduced DUB activity and indicate that deubiquitylation is central to CI induction in cid systems. We also characterize effects of other genetic variation distinguishing wMel-like cifs. Importantly, CI strength determines Wolbachia prevalence in natural systems and directly influences the efficacy of Wolbachia biocontrol strategies in transinfected mosquito systems. These approaches rely on strong CI to reduce human disease.All study data are included in the article and/or supporting information.
Discrete dynamical models on Wolbachia infection frequency in mosquito populations with biased release ratios
18717Y. Shi and B. Zheng, Journal of Biological Dynamics, 2021-09-18 14:11:29.
We develop two discrete models to study how supplemental releases affect the Wolbachia spreading dynamics in cage mosquito populations. The first model focuses on the case when only infected males are released at each generation. This release strategy has been proved to be capable of speeding up the Wolbachia persistence by suppressing the compatible matings between uninfected individuals. The second model targets the case when only infected females are released at each generation. For both models, detailed model formulation, enumeration of the positive equilibria and their stability analysis are provided. Theoretical results show that the two models can generate bistable dynamics when there are three positive equilibrium points, semi-stable dynamics for the case of two positive equilibrium points. And when the positive equilibrium point is unique, it is globally asymptotically stable. Some numerical simulations are offered to get helpful implications on the design of the release strategy.
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.
Diverse wMel variants of Wolbachia pipientis differentially rescue fertility and cytological defects of the bag of marbles partial loss of function mutation in Drosophila melanogaster
18817J. E. Bubnell, P. Fernandez-Begne, C. K. S. Ulbing and C. F. Aquadro, G3 Genes|Genomes|Genetics, 2021-09-07 15:29:22.
In Drosophila melanogaster, the maternally inherited endosymbiont Wolbachia pipientis interacts with germline stem cell genes during oogenesis. One such gene, bag of marbles (bam) is the key switch for differentiation and also shows signals of adaptive evolution for protein diversification. These observations have led us to hypothesize that W. pipientis could be driving the adaptive evolution of bam for control of oogenesis. To test this hypothesis, we must understand the specificity of the genetic interaction between bam and W. pipientis. Previously, we documented that the W. pipientis variant, wMel, rescued the fertility of the bamBW hypomorphic mutant as a transheterozygote over a bam null. However, bamBW was generated more than 20 years ago in an uncontrolled genetic background and maintained over a balancer chromosome. Consequently, the chromosome carrying bamBW accumulated mutations that have prevented controlled experiments to further assess the interaction. Here, we used CRISPR/Cas9 to engineer the same single amino acid bam hypomorphic mutation (bamL255F) and a new bam null disruption mutation into the w1118 isogenic background. We assess the fertility of wildtype bam, bamL255F/bamnull hypomorphic, and bamL255F/bamL255F mutant females, each infected individually with 10 W. pipientis wMel variants representing three phylogenetic clades. Overall, we find that all of the W. pipientis variants tested here rescue bam hypomorphic fertility defects with wMelCS-like variants exhibiting the strongest rescue effects. In addition, these variants did not increase wildtype bam female fertility. Therefore, both bam and W. pipientis interact in genotype-specific ways to modulate female fertility, a critical fitness phenotype.
Millions of Lab-Grown Mosquitoes Are Being Released in Guangzhou
18219F. Yiying, Sixth Tone, 2021-08-25 15:26:45.
Guangzhou is releasing millions of lab-engineered mosquitoes every day to neuter and prevent preexisting mosquitoes in the environment from spreading vector-borne diseases, local television station reported Saturday.The Guangzhou Wolbaki Biotech Co., Ltd., in partnership with the city’s disease control and prevention bureau, releases about five million lab-grown male mosquitoes daily, which only mate with aedes albopictus mosquitoes to stop them from reproducing disease-carrying offspring, according to the media report. Only female aedes albopictus, also known as tiger mosquitoes, prey on people, potentially transmitting the viruses that cause dengue and chikungunya, among other illnesses.The biotech company’s “mosquito factory” in Guangzhou — where dengue fever is a public health concern — breeds mosquitoes to produce offspring carrying the Wolbachia bacteria, which is commonly found in insects and not harmful to humans, before releasing them to the environment. When mosquitoes carrying Wolbachia mate with aedes albopictus, it reduces the replication of the viruses they carry, making them less likely to transmit it to humans.
Sex separation of Aedes spp. mosquitoes for sterile insect technique application: a review
17905B. M. Moran-Aceves, C. F. Marina, A. Dor, P. Liedo and J. Toledo, Entomologia Experimentalis Et Applicata, 10. 2021-07-24 15:04:30.
Separation of the sexes is necessary for the application of the sterile insect technique (SIT) in mosquitoes due to the hematophagous habits and disease vector activity of the females. In this review we analyze the history, current status, and future perspectives for the development of genetic sexing strains (GSS) of Aedes mosquitoes (Diptera: Culicidae). Various genetic control methods for mosquitoes are reviewed, as are their need for sex-separation methods. We focus on areas of opportunity where GSS developed with classical genetic methods can be used. Regulatory restrictions and social acceptance of various control methods are analyzed. We conclude that the development of GSS by classical methods represents the most viable option for separation of the sexes and the application of large-scale SIT programs within an area-wide integrated vector management (AW-IVM) approach.
Wolbachia as translational science: controlling mosquito-borne pathogens
17799E. P. Caragata, H. L. C. Dutra, P. H. F. Sucupira, A. G. A. Ferreira and L. A. Moreira, Trends in Parasitology, 2021-07-22 20:54:55.
In this review we examine how exploiting the Wolbachia?mosquito relationship has become an increasingly popular strategy for controlling arbovirus transmission. Field deployments of Wolbachia-infected mosquitoes have led to significant decreases in dengue virus incidence via high levels of mosquito population suppression and replacement, emphasizing the success of Wolbachia approaches. Here, we examine how improved knowledge of Wolbachia?host interactions has provided key insight into the mechanisms of the essential phenotypes of pathogen blocking and cytoplasmic incompatibility. And we discuss recent studies demonstrating that extrinsic factors, such as ambient temperature, can modulate Wolbachia density and maternal transmission. Finally, we assess the prospects of using Wolbachia to control other vectors and agricultural pest species.
Manipulated Mosquitoes Cut Dengue by 77%
17513T. Hayes, Healthcare Packaging, 2021-06-22 14:34:21.
Dengue, a mosquito-borne viral disease, wasn’t that common 50 years ago. In fact, only nine countries had severe outbreaks. But since then, it’s been on a steady incline to the point that there are now 400 million infections a year that contribute to 22,000 deaths. Thankfully, we have the World Mosquito Programme, a group that’s fighting the tropical mosquito disease head-on with genetically modified mosquitoes. The insects are infected with Wolbachia, a bacteria that inhibits viruses’ ability to live inside the insects, and also controls reproduction so that offspring are also infected with the bacteria. The result is a new population of insects that can’t transmit viruses like dengue, yellow fever and Zika. The study in Indonesia included 8,000 people, and concluded with protective efficacy of 77.1%.
Dengue Infections Can Be Sharply Reduced With Wolbachia Bacteria
17453J. Stone, Medscape, 2021-06-14 17:25:12.
A modestly titled new study released in the New England Journal of Medicine belies the extraordinary 77% protective efficacy reported for preventing dengue infections with Wolbachia-infected Aedes aegypti mosquitoes. A cluster-randomized clinical trial, the AWED ("Applying Wolbachia to Eliminate Dengue") study was conducted in Yogyakarta, Indonesia and led by professors Adi Utarini, MD, PhD, of Gadjah Mada University and Cameron Simmons, PhD, the World Mosquito Program's Oceana director, in partnership with the Tahija Foundation and Monash University.
Making mosquitoes to fight mosquitoes to prevent dengue
17484A. George, Times of India, 2021-06-14 15:04:01.
In 2017, Delhi Chief Minister Arvind Kejriwal tweeted that the national capitl needed to be made mosquito-free. The same year, his Kerala counterpart, Pinaray Vijayan, called a three-day state-wide cleanliness drive as hospitals filled with genue patients.
Stable high-density and maternally inherited Wolbachia infections in Anopheles moucheti and Anopheles demeilloni mosquitoes
17504T. Walker, S. Quek, C. L. Jeffries, J. Bandibabone, V. Dhokiya, R. Bamou, M. Kristan, L. A. Messenger, A. Gidley, E. A. Hornett, E. R. Anderson, C. Cansado-Utrilla, S. Hegde, C. Bantuzeko, J. C. Stevenson, N. F. Lobo, S. C. Wagstaff, C. A. Nkondjio, S. R., Current Biology, 31:2310. 2021-06-14 13:55:55.
Wolbachia, a widespread bacterium that can reduce pathogen transmission in mosquitoes, has recently been reported to be present in Anopheles (An.) species. In wild populations of the An. gambiae complex, the primary vectors of Plasmodium malaria in Sub-Saharan Africa, Wolbachia DNA sequences at low density and infection frequencies have been detected. As the majority of studies have used highly sensitive nested PCR as the only method of detection, more robust evidence is required to determine whether Wolbachia strains are established as endosymbionts in Anopheles species. Here, we describe high-density Wolbachia infections in geographically diverse populations of An. moucheti and An. demeilloni. Fluorescent in situ hybridization localized a heavy infection in the ovaries of An. moucheti, and maternal transmission was observed. Genome sequencing of both Wolbachia strains obtained genome depths and coverages comparable to those of other known infections. Notably, homologs of cytoplasmic incompatibility factor (cif) genes were present, indicating that these strains possess the capacity to induce the cytoplasmic incompatibility phenotype, which allows Wolbachia to spread through host populations. These strains should be further investigated as candidates for use in Wolbachia biocontrol strategies in Anopheles aiming to reduce the transmission of malaria.
Dengue Fever Cut Down by 77% With Groundbreaking Bacteria-Armed Mosquitoes
17322M. Davis, The Science Times, 2021-06-10 12:02:22.
Scientists found that dengue fever cases have decreased by 77% in a groundbreaking trial that took place in Yogyakarta City, Indonesia. They used Wolbachia-infected mosquitoes that reduced their ability to spread the dengue fever. The team at the World Mosquito Program said that this could be a solution to dengue fever that is prevalent around the world. Dr. Katie Anders, one of the researchers, described the trial of using Wolbachia-infected mosquitoes in Indonesia as a "naturally miraculous" solution. Wolbachia is a virus-fighting bacteria that has been approved since 2017 by the Environmental Protection Agency (EPA) for use in the United States. Other interventions, like genetically modified mosquitoes, are also used to combat dengue fever and other diseases caused by mosquitoes. According to BBC News, Wolbachia does not harm the mosquito, but it will only be stored in the same parts of the mosquito's body where the dengue virus is kept. It competes for the resources, preventing the dengue virus to replicate, and therefore the mosquito is less likely to cause the infection when it bites. Moreover, Wolbachia bacteria can also manipulate and alter the fertility of their hosts. They make sure that they are passed on to the next generation of mosquitoes, which means they are established and should stick around for a long time to continue their protective abilities against dengue fever.
Mosquito ‘bacteria hack’ nearly eliminates dengue fever and could save millions of lives
17316A. Wilkins, METRO, 2021-06-10 11:50:12.
Mosquitoes infected with a ‘miraculous’ bacteria have been shown to reduce dengue fever cases by 77%, in a groundbreaking new study. Scientists released mosquitoes infected with ‘Wolbachia’ bacteria into the Indonesian city of Yogyakarta – but only in certain zones. In the zones where the modified mosquitoes had been released, cases of dengue fell by 77% and hospitalisations dropped by 86%. The results of the study, carried out by the World Mosquito Programme (WMP), were ‘better than we could have hoped for’, according to researcher Dr. Katie Anders. Cases of dengue fever, a virus that can cause muscle and bone pain, and death, have risen rapidly in the past decades – there are an estimated 400 million cases a year. The Wolbachia bacteria used in the trial were chosen because it hides in the same parts of a mosquito’s body that dengue virus hides in. A bacteria that is benign to humans, the Wolbachia then competes for resources with the dengue virus, which makes it less likely for the mosquito to cause a dengue infection when it bites a human.
‘Miraculous’ mosquito hack cuts dengue by 77%
17311J. Gallagher, BBC, 2021-06-10 11:38:57.
Dengue fever cases have been cut by 77% in a "groundbreaking" trial that manipulates the mosquitoes that spread it, say scientists. They used mosquitoes infected with "miraculous" bacteria that reduce the insect's ability to spread dengue. The trial took place in Yogyakarta city, Indonesia, and is being expanded in the hope of eradicating the virus. The World Mosquito Programme team says it could be a solution to a virus that has gone around the world. Few people had heard of dengue 50 years ago, but it has been a relentless slow-burning pandemic and cases have increased dramatically. In 1970, only nine countries had faced severe dengue outbreaks, now there are up to 400 million infections a year. Dengue is commonly known as "break-bone fever" because it causes severe pain in muscles and bones and explosive outbreaks can overwhelm hospitals.
Modified mosquitoes reduce dengue cases by 77% in Indonesia experiment
17309M. Fox, CNN, 2021-06-10 11:32:59.
An experiment to infect mosquitoes with bacteria that stop them from transmitting viruses appears to have helped reduced the spread of deadly dengue virus in Indonesia, researchers reported Wednesday. The modified mosquitoes thrived for three years, and cases of dengue were reduced by 77% in areas where they were introduced, the researchers reported in the New England Journal of Medicine. The mosquitoes are infected with bacteria called Wolbachia, which not only interfere with the ability of viruses to live in the bodies of the insects, but which also control reproduction so that the mosquitoes only have Wolbachia-infected offspring. The result is a growing population of insects that don't pass on viruses such as dengue, yellow fever and Zika. The study involved more than 8,000 people, about half of whom lived in areas where the modified Aedes aegypti mosquitoes had been living and breeding.
Efficacy of Wolbachia-Infected Mosquito Deployments for the Control of Dengue
17300A. Utarini, C. Indriani, R. A. Ahmad, W. Tantowijoyo, E. Arguni, M. R. Ansari, E. Supriyati, D. S. Wardana, Y. Meitika, I. Ernesia, I. Nurhayati, E. Prabowo, B. Andari, B. R. Green, L. Hodgson, Z. Cutcher, E. Rancès, P. A. Ryan, S. L. O’Neill, S. M. Dufau, New England Journal of Medicine, 384:2177-2186. 2021-06-10 11:05:41.
BACKGROUND Aedes aegypti mosquitoes infected with the wMel strain of Wolbachia pipientis are less susceptible than wild-type A. aegypti to dengue virus infection. METHODS We conducted a cluster-randomized trial involving releases of wMel-infected A. aegypti mosquitoes for the control of dengue in Yogyakarta, Indonesia. We randomly assigned 12 geographic clusters to receive deployments of wMel-infected A. aegypti (intervention clusters) and 12 clusters to receive no deployments (control clusters). All clusters practiced local mosquito-control measures as usual. A test-negative design was used to assess the efficacy of the intervention. Patients with acute undifferentiated fever who presented to local primary care clinics and were 3 to 45 years of age were recruited. Laboratory testing was used to identify participants who had virologically confirmed dengue (VCD) and those who were test-negative controls. The primary end point was symptomatic VCD of any severity caused by any dengue virus serotype. RESULTS After successful introgression of wMel into the intervention clusters, 8144 participants were enrolled; 3721 lived in intervention clusters, and 4423 lived in control clusters. In the intention-to-treat analysis, VCD occurred in 67 of 2905 participants (2.3%) in the intervention clusters and in 318 of 3401 (9.4%) in the control clusters (aggregate odds ratio for VCD, 0.23; 95% confidence interval [CI], 0.15 to 0.35; P=0.004). The protective efficacy of the intervention was 77.1% (95% CI, 65.3 to 84.9) and was similar against the four dengue virus serotypes. The incidence of hospitalization for VCD was lower among participants who lived in intervention clusters (13 of 2905 participants [0.4%]) than among those who lived in control clusters (102 of 3401 [3.0%]) (protective efficacy, 86.2%; 95% CI, 66.2 to 94.3).
A Pivotal Mosquito Experiment Could Not Have Gone Better
17294E. Yong, The Atlantic, 2021-06-10 10:50:58.
The World Mosquito Program (WMP), a nonprofit that pioneered this technique, had run small pilot studies in Australia that suggested it could work. Utarini, who co-leads WMP Yogyakarta, has now shown conclusively that it does. Her team released Wolbachia-carrying mosquitoes in parts of Yogyakarta as part of a randomized controlled trial. The results, which were unveiled last year and have now been published, showed that Wolbachia rapidly spread among the local mosquitoes, and reduced the incidence of dengue by 77 percent. “That provides the gold standard of evidence that Wolbachia is a highly effective intervention against dengue,” says Oliver Brady, a dengue expert at the London School of Hygiene and Tropical Medicine, who was not involved in the study. “It has the potential to revolutionize mosquito control.” The trial’s results were so encouraging that the researchers have since released Wolbachia-carrying mosquitoes over all of central Yogyakarta—a 32-square-kilometer zone that’s home to more than 400,000 people. They’re now expanding into the densest surrounding provinces, aiming to protect 4 million people by the end of 2022. If they succeed, they should be able to prevent more than 10,000 dengue infections every year, Katherine Anders of the WMP told me. And the team is optimistic enough that it’s daring to think about an even grander goal: eliminating dengue from the city altogether.
Mosquitoes armed with virus-fighting bacteria sharply curb dengue infections, hospitalizations
17319K. Servick, Science, 2021-06-09 11:55:26.
A strategy for fighting dengue fever with bacteria-armed mosquitoes has passed its most rigorous test yet: a large, randomized, controlled trial. Researchers reported today dramatic reductions in rates of dengue infection and hospitalization in areas of an Indonesian city where the disease-fighting mosquitoes were released. The team expects the World Health Organization (WHO) to formally recommend the approach for broader use. The findings are a “breakthrough” that brings the approach “much closer to … being an official strategy to control dengue,” says Ewa Chrostek, an infection biologist at the University of Liverpool who was not involved with the work. WHO estimates there are 100 million to 400 million infections per year with dengue, which can cause high fever and severe joint pain. The bacterium Wolbachia pipientis naturally inhabits many insects, though not Aedes aegypti mosquitoes, the main transmitter of dengue virus. In A. aegypti cells, the bacterium can block viruses, including dengue, from replicating, making the insects less likely to spread disease when they bite humans. That has made the microbe a promising strategy for fighting dengue. In tropical regions, where mosquito-borne viruses are common, other strategies such as insecticides have failed to fully control the disease.
Study demonstrates ‘exciting potential’ of Wolbachia-infected mosquitoes to control dengue
17313G. Gallagher, Healio, 2021-06-09 11:43:08.
The release of Wolbachia-infected mosquitoes led to a 77% reduction in the incidence of symptomatic dengue in an Indonesian city, according to researchers, who said the same approach could be used to fight other mosquito-borne diseases. The study tested a strain of Wolbachia pipientis called wMel that makes Aedes aegypti mosquitoes less susceptible to dengue virus infection. Wolbachia pipientis occurs naturally in many insects but not A. aegypti, the primary vector of dengue, according to Adi Utarini, PhD, MPH, MSc, and colleagues from the nonprofit World Mosquito Program, which has been releasing Wolbachia-infected mosquitoes in the wild since 2011. A study in Indonesia demonstrated the potential of a natural intervention to significantly reduce the incidence of dengue, the world’s most prevalent mosquito-borne disease. “Wolbachia facilitates its own population introgression by manipulating reproductive outcomes between wild-type and Wolbachia-infected mosquitoes: the only viable mating outcomes are those in which the progeny are infected with Wolbachia,” they wrote in the new report, which was published Wednesday in The New England Journal of Medicine.
Vector control: Discovery of Wolbachia in malaria vectors
17554P. A. Ross and A. A. Hoffmann, Current Biology, 31:R738-R740. 2021-06-07 14:28:04.
Wolbachia bacteria are being widely released for suppression of dengue transmitted by Aedes mosquitoes. Walker, Quek, Jeffries and colleagues present robust evidence for natural Wolbachia infections in malaria-vectoring Anopheles mosquitoes, paving the way for new Wolbachia-based intervention
Living in the endosymbiotic world of Wolbachia: A centennial review
17794R. Kaur, J. D. Shropshire, K. L. Cross, B. Leigh, A. J. Mansueto, V. Stewart, S. R. Bordenstein and S. R. Bordenstein, Cell Host and Microbe, 29:879-893. 2021-05-03 20:31:30.
The most widespread intracellular bacteria in the animal kingdom are maternally inherited endosymbionts of the genus Wolbachia. Their prevalence in arthropods and nematodes worldwide and stunning arsenal of parasitic and mutualistic adaptations make these bacteria a biological archetype for basic studies of symbiosis and applied outcomes for curbing human and agricultural diseases. Here, we conduct a summative, centennial analysis of living in the Wolbachia world. We synthesize literature on Wolbachia's host range, phylogenetic diversity, genomics, cell biology, and applications to filarial, arboviral, and agricultural diseases. We also review the mobilome of Wolbachia including phage WO and its essentiality to hallmark reproductive phenotypes in arthropods. Finally, the Wolbachia system is an exemplar for discovery-based science education using biodiversity, biotechnology, and bioinformatics lessons. As we approach a century of Wolbachia research, the interdisciplinary science of this symbiosis stands as a model for consolidating and teaching the integrative rules of endosymbiotic life.
Mechanistically comparing reproductive manipulations caused by selfish chromosomes and bacterial symbionts
16958E. Dalla Benetta, O. S. Akbari and P. M. Ferree, Heredity, 126:707-716. 2021-05-01 14:52:23.
Insects naturally harbor a broad range of selfish agents that can manipulate their reproduction and development, often leading to host sex ratio distortion. Such effects directly benefit the spread of the selfish agents. These agents include two broad groups: bacterial symbionts and selfish chromosomes. Recent studies have made steady progress in uncovering the cellular targets of these agents and their effector genes. Here we highlight what is known about the targeted developmental processes, developmental timing, and effector genes expressed by several selfish agents. It is now becoming apparent that: (1) the genetic toolkits used by these agents to induce a given reproductive manipulation are simple, (2) these agents target sex-specific cellular processes very early in development, and (3) in some cases, similar processes are targeted. Knowledge of the molecular underpinnings of these systems will help to solve long-standing puzzles and provide new tools for controlling insect pests.
When More is Less: Mosquito Population Suppression Using Sterile, Incompatible and Genetically Modified Male Mosquitoes
18708S. L. Dobson, Journal of Medical Entomology, 58:1980-1986. 2021-03-11 13:50:08.
The current review of the Sterile Insect Technique (SIT) is motivated by new technologies and the recent renaissance of male release field trials, which is driving an evolution in mosquito control and regulation. Practitioners that are releasing male mosquitoes would do well to learn from past successes and failures, including political and public engagement complications. With examples that include nuanced integrations of the different technologies, e.g., combinations of Wolbachia and irradiation, it is critical that scientists understand and communicate accurately about the technologies, including their evolving management by different regulatory agencies in the USA. Some male release approaches are considered ‘pesticides’ and regulated by federal and state agencies, while other male release approaches are unregulated. It is important to consider how the new technologies fit with the more ‘traditional’ chemical applications of adulticides and larvicides. The economics of male release programs are substantially different from traditional control costs, which can be a challenge to their adoption by abatement districts. However, there is substantial need to overcome these complications and challenges, because the problem with invasive mosquitoes grows ever worse with factors that include insecticide resistance, globalization and climate change.
How to engage communities on a large scale? Lessons from World Mosquito Program in Rio de Janeiro, Brazil [version 2; peer review: 1 approved, 2 approved with reservations]
16361G. B. Costa, R. Smithyman, S. L. O'Neill and L. A. Moreira, Gates Open Research, 2021-02-04 19:26:09.
Here we discuss and analyse the framework for community engagement implemented by the WMP in Brazil, during the large-scale deployment of the method in the municipalities of Niterói and Rio de Janeiro, Brazil. Our experience indicates that the community engagement work for arboviruses control should be understood as an opportunity for local development. It is necessary, based on an integrated analysis of the territory, to understand that the actions for arboviruses control could be a catalyst for the necessary socioenvironmental, cultural and public health changes. Furthermore, it is essential to understand that community engagement goes beyond informing or asking for population consent, but it constitutes a possibility for dialogue and exchange between the various stakeholders present in the territories, to build on cooperation for mosquito-borne disease control.
Project Wolbachia: Residents are killing the ‘helpful’ mosquitoes, which can be a nuisance
16228T. J. Cheng, today, 2021-01-29 16:33:12.
In 2019, Dr Amy Khor, then Senior Minister of State for the Environment and Water Resources, said that there was a 90 per cent suppression rate at study sites in Tampines and Yishun from February to November that year. However, certain public housing estates under the project were still dengue hot zones last year.The idea of the project is to have the male Aedes mosquitoes, which are injected with the Wolbachia bacteria, mate with female Aedes aegypti mosquitoes. The females then go on to lay eggs that do not hatch, effectively suppressing their numbers.
Next-generation tools to control biting midge populations and reduce pathogen transmission
15940P. Shults, L. W. Cohnstaedt, Z. N. Adelman and C. Brelsfoard, Parasites and Vectors, 14:31. 2021-01-07 14:52:23.
Biting midges of the genus Culicoides transmit disease-causing agents resulting in a significant economic impact on livestock industries in many parts of the world. Localized control efforts, such as removal of larval habitat or pesticide application, can be logistically difficult, expensive and ineffective if not instituted and maintained properly. With these limitations, a population-level approach to the management of Culicoides midges should be investigated as a means to replace or supplement existing control strategies. Next-generation control methods such as Wolbachia- and genetic-based population suppression and replacement are being investigated in several vector species. Here we assess the feasibility and applicability of these approaches for use against biting midges. We also discuss the technical and logistical hurdles needing to be addressed for each method to be successful, as well as emphasize the importance of addressing community engagement and involving stakeholders in the investigation and development of these approaches.
Wolbachia strain wAlbB maintains high density and dengue inhibition following introduction into a field population of Aedes aegypti
15689N. A. Ahmad, M.-V. Mancini, T. H. Ant, J. Martinez, G. M. R. Kamarul, W. A. Nazni, A. A. Hoffmann and S. P. Sinkins, Philosophical Transactions of the Royal Society B: Biological Sciences, 376:20190809. 2020-12-28 15:06:28.
Here, wAlbB-carrying Ae. aegypti collected from the field 20 months after the cessation of releases showed no reduction in Wolbachia density or tissue distribution changes compared to a wAlbB laboratory colony. The wAlbB strain continued to induce complete unidirectional cytoplasmic incompatibility, showed perfect maternal transmission under laboratory conditions, and retained its capacity to inhibit dengue. Additionally, a field-collected wAlbB line was challenged with Malaysian dengue patient blood, and showed significant blocking of virus dissemination to the salivary glands.
New insect species made via genetic engineering
15610L. Leffer, SCIENCELINE, 2020-12-18 15:47:16.
A biotech fast-forward button for evolution is on the horizon. Researchers say they have used a novel genetic engineering method to create several new species of fruit fly in the lab for the first time — an achievement which might help put a future without malaria and other insect-borne diseases within reach. The approach, called synthetic speciation, could prove useful in creating safer pest-control technologies, says Maciej Maselko, a postdoctoral fellow studying synthetic biology at Macquarie University. In one far-off scenario, according to Maselko, synthetic speciation might even be applied to generate designer organisms that could pollinate plants or even detect landmines. Maselko and his team published their findings September 8 in Nature Communications. “Speciation has occurred billions of times on the planet, but hasn’t been engineerable [before],” says Michael Smanski, a molecular biologist at the University of Minnesota and member of the research team. Maselko, Smanski and their colleagues have previously used a similar method to engineer “species like” differences in yeast in 2018, but their more recent results are the first time the concept has been proven possible in a multicellular animal. This method could produce untold numbers of new animal varieties within months rather than millennia, Smanski says.
‘A plague to be reckoned with’: UMN research creates a buzz with invasive fruit fly research
15238B. Most, The Minnesota Daily, 2020-11-18 13:26:31.
n early November, assistant professor Mike Smanski published an article about a new breakthrough in this research, demonstrating for the first time this kind of genetic engineering was possible in the common fruit fly. This shows that researchers could engineer this work into spotted wing drosophila in the future. The University’s Smanski Lab has also studied this technique in mosquitoes, zebra fish and carp, but never with this type of fruit fly, he said. “These are all a new class of genetic pesticide, basically, that allow you to engineer the pest organism itself and convert that pest organism into the pesticide,” Smanski said. Through this work the researchers can create a pest that is biologically the same, but when the females mate with these genetically modified males, they will not produce viable offspring, he said. This sterile insect technique can be helpful not only in reducing the population of insects, but in reducing the impacts of insecticides on surrounding species and nearby ecosystems, said Feltman, a second-year biochemistry, molecular biology and biophysics graduate student.
Vector-Focused Approaches to Curb Malaria Transmission in the Brazilian Amazon: An Overview of Current and Future Challenges and Strategies
15960E. M. Rocha, R. D. Katak, J. C. de Oliveira, M. D. Araujo, B. C. Carlos, R. Galizi, F. Tripet, O. Marinotti and J. A. Souza, Tropical Medicine and Infectious Disease, 5. 2020-10-20 18:16:39.
Here we present an overview on both conventional and novel promising vector-focused tools to curb malaria transmission in the Brazilian Amazon. If well designed and employed, vector-based approaches may improve the implementation of malaria-control programs, particularly in remote or difficult-to-access areas and in regions where existing interventions have been unable to eliminate disease transmission. However, much effort still has to be put into research expanding the knowledge of neotropical malaria vectors to set the steppingstones for the optimization of conventional and development of innovative vector-control tools.
Stable Introduction of Plant-Virus-Inhibiting Wolbachia into Planthoppers for Rice Protection
19719J. T. Gong, Y. Li, T. P. Li, Y. Liang, L. Hu, D. Zhang, C. Y. Zhou, C. Yang, X. Zhang, S. S. Zha, X. Z. Duan, L. A. Baton, X. Y. Hong, A. A. Hoffmann and Z. Xi, Current Biology, 30:4837-4845.e5. 2020-10-10 12:20:07.
Progress has been made in developing the maternally inherited endosymbiotic bacterium Wolbachia as a tool for protecting humans from mosquito-borne diseases. In contrast, Wolbachia-based approaches have not yet been developed for the protection of plants from insect pests and their associated diseases, with a major challenge being the establishment of artificial Wolbachia infections expressing desired characteristics in the hemipterans that transmit the majority of plant viruses. Here, we report stable introduction of Wolbachia into the brown planthopper, Nilaparvata lugens, the most destructive rice pest that annually destroys millions of hectares of staple crops. The Wolbachia strain wStri from the small brown planthopper, Laodelphax striatellus, was transferred to this new host, where it showed high levels of cytoplasmic incompatibility, enabling rapid invasion of laboratory populations. Furthermore, wStri inhibited infection and transmission of Rice ragged stunt virus and mitigated virus-induced symptoms in rice plants, opening up the development of Wolbachia-based strategies against major agricultural pests and their transmitted pathogens
Cytoplasmic incompatibility: an autocidal mechanism for mosquito population control
13893V. Dev, BugBitten BMC, 2020-08-18 13:23:13.
Cytoplasmic incompatibility resulting in non-reciprocal fertility is a naturally occurring phenomenon, but remains unexplored to greater extent for the control of insect vector populations. This mechanism deserves priority for mosquito control and reducing disease transmission, being non-insecticidal and easier to operate with minimal investments.
Modeling the suppression dynamics of Aedes mosquitoes with mating inhomogeneity
13847M. Huang and L. Hu, Journal of Biological Dynamics, 14:656-678. 2020-08-04 13:51:41.
In this work, we introduce a delay differential equation model with mating inhomogeneity to discuss mosquito population suppression based on Wolbachia. Our analyses show that the wild mosquitoes could be eliminated if either the adult mortality rate exceeds the threshold δ∗A or the release amount exceeds the threshold r∗ uniformly. Our simulations suggest that the releasing should be started at least 5 weeks before the peak dengue season, taking into account both the release amount and the suppression speed.
The Biochemistry of Cytoplasmic Incompatibility Caused by Endosymbiotic Bacteria
18757H. Chen, M. Zhang and M. Hochstrasser, Genes, 11. 2020-07-25 14:17:05.
Many species of arthropods carry maternally inherited bacterial endosymbionts that can influence host sexual reproduction to benefit the bacterium. The most well-known of such reproductive parasites is Wolbachia pipientis. Wolbachia are obligate intracellular α-proteobacteria found in nearly half of all arthropod species. This success has been attributed in part to their ability to manipulate host reproduction to favor infected females. Cytoplasmic incompatibility (CI), a phenomenon wherein Wolbachia infection renders males sterile when they mate with uninfected females, but not infected females (the rescue mating), appears to be the most common. CI provides a reproductive advantage to infected females in the presence of a threshold level of infected males. The molecular mechanisms of CI and other reproductive manipulations, such as male killing, parthenogenesis, and feminization, have remained mysterious for many decades. It had been proposed by Werren more than two decades ago that CI is caused by a Wolbachia-mediated sperm modification and that rescue is achieved by a Wolbachia-encoded rescue factor in the infected egg. In the past few years, new research has highlighted a set of syntenic Wolbachia gene pairs encoding CI-inducing factors (Cifs) as the key players for the induction of CI and its rescue. Within each Cif pair, the protein encoded by the upstream gene is denoted A and the downstream gene B. To date, two types of Cifs have been characterized based on the enzymatic activity identified in the B protein of each protein pair; one type encodes a deubiquitylase (thus named CI-inducing deubiquitylase or cid), and a second type encodes a nuclease (named CI-inducing nuclease or cin). The CidA and CinA proteins bind tightly and specifically to their respective CidB and CinB partners. In transgenic Drosophila melanogaster, the expression of either the Cid or Cin protein pair in the male germline induces CI and the expression of the cognate A protein in females is sufficient for rescue. With the identity of the Wolbachia CI induction and rescue factors now known, research in the field has turned to directed studies on the molecular mechanisms of CI, which we review here.
Genetic Biocontrol – An Overview (video 13.13 min)
11398GeneConvene Global Collaborative, 2020-04-25 12:41:00.
This video explains what genetic biocontrol is and surveys various technologies that can be consider genetic biocontrol technologies. It offers a conceptual organization of the various technologies based on the potential of genetic biocontrol organisms to persist and spread in an environment. This video was produced by the GeneConvene Global Colaborative.
Wolbachia transinfections in Culex quinquefasciatus generate cytoplasmic incompatibility
6224T. H. Ant, C. Herd, F. Louis, A. B. Failloux and S. P. Sinkins, Insect Molecular Biology, 29:1-8. 2020-01-06 21:43:12.
Culex quinquefasciatus is an important mosquito vector of a number of viral and protozoan pathogens of humans and animals, and naturally carries the endosymbiont Wolbachia pipientis, strain wPip. Wolbachia are used in two distinct vector control strategies: firstly, population suppression caused by mating incompatibilities between mass-released transinfected males and wild females; and secondly, the spread of pathogen transmission-blocking strains through populations. Using embryonic microinjection, two novel Wolbachia transinfections were generated in C. quinquefasciatus using strains native to the mosquito Aedes albopictus: a wAlbB single infection, and a wPip plus wAlbA superinfection. The wAlbB infection showed full bidirectional cytoplasmic incompatibility (CI) with wild-type C. quinquefasciatus in reciprocal crosses. The wPipwAlbA superinfection showed complete unidirectional CI, and therefore population invasion potential. Whereas the wAlbB strain showed comparatively low overall densities, similar to the native wPip, the wPipwAlbA superinfection reached over 400-fold higher densities in the salivary glands compared to the native wPip, suggesting it may be a candidate for pathogen transmission blocking.
Genetic control of Invasive carp
13941MAISRC, Minnesota Aquatic Invasive Species Research Center, 2019-10-16 15:08:53.
This project focuses on a novel method of biocontrol for common carp which will complement existing technologies by introducing a synthetic species-like barrier to reproduction. Researchers will use programmable transcription activators to drive lethal embryonic overexpression of endogenous genes in hybrid embryos.
The toxin–antidote model of cytoplasmic incompatibility: Genetics and evolutionary implications
3889Beckmann, JFB, Manon; Chen, Hongli; Hochstrasser, Mark; Poinsot, Denis; Merçot, Hervé; Weill, Mylène; Sicard, Mathieu; Charlat, Sylvain, Trends in Genetics, 35:175-185. 2019-01-07 00:00:00.
Wolbachia bacteria inhabit the cells of about half of all arthropod species, an unparalleled success stemming in large part from selfish invasive strategies. Cytoplasmic incompatibility (CI), whereby the symbiont makes itself essential to embryo viability, is the most common of these and constitutes a promising weapon against vector-borne diseases. After decades of theoretical and experimental struggle, major recent advances have been made toward a molecular understanding of this phenomenon. As pieces of the puzzle come together, from yeast and Drosophila fly transgenesis to CI diversity patterns in natural mosquito populations, it becomes clearer than ever that the CI induction and rescue stem from a toxin–antidote (TA) system. Further, the tight association of the CI genes with prophages provides clues to the possible evolutionary origin of this phenomenon and the levels of selection at play.
One prophage WO gene rescues cytoplasmic incompatibility in Drosophila melanogaster
5926Shropshire, J. D., J. On, E. M. Layton, H. Zhou and S. R. Bordenstein, Proceedings of the National Academy of Sciences, 115:4987. 2018-04-23 16:58:30.
The World Health Organization recommended pilot deployment of Wolbachia-infected mosquitoes to curb viral transmission to humans. Releases of mosquitoes are underway worldwide because Wolbachia can block replication of these pathogenic viruses and deterministically spread by a drive system termed cytoplasmic incompatibility (CI). Despite extensive research, the underlying genetic basis of CI remains only half-solved. We recently reported that two prophage WO genes recapitulate the modification component of CI in a released strain for vector control. Here we show that one of these genes underpins rescue of CI. Together, our results reveal the complete genetic basis of this selfish trait and pave the way for future studies exploring WO prophage genes as adjuncts or alternatives to current control efforts.Wolbachia are maternally inherited, intracellular bacteria at the forefront of vector control efforts to curb arbovirus transmission. In international field trials, the cytoplasmic incompatibility (CI) drive system of wMel Wolbachia is deployed to replace target vector populations, whereby a Wolbachia-induced modification of the sperm genome kills embryos. However, Wolbachia in the embryo rescue the sperm genome impairment, and therefore CI results in a strong fitness advantage for infected females that transmit the bacteria to offspring. The two genes responsible for the wMel-induced sperm modification of CI, cifA and cifB, were recently identified in the eukaryotic association module of prophage WO, but the genetic basis of rescue is unresolved. Here we use transgenic and cytological approaches to demonstrate that maternal cifA expression independently rescues CI and nullifies embryonic death caused by wMel Wolbachia in Drosophila melanogaster. Discovery of cifA as the rescue gene and previously one of two CI induction genes establishes a “Two-by-One” model that underpins the genetic basis of CI. Results highlight the central role of prophage WO in shaping Wolbachia phenotypes that are significant to arthropod evolution and vector control.
Evolutionary genetics of cytoplasmic incompatibility genes cifA and cifB in prophage WO of Wolbachia
5929Lindsey, A. R. I., D. W. Rice, S. R. Bordenstein, A. W. Brooks, S. R. Bordenstein and I. L. G. Newton, Genome Biology and Evolution, 10:434-451. 2018-01-17 17:04:14.
The bacterial endosymbiont Wolbachia manipulates arthropod reproduction to facilitate its maternal spread through host populations. The most common manipulation is cytoplasmic incompatibility (CI): Wolbachia-infected males produce modified sperm that cause embryonic mortality, unless rescued by embryos harboring the same Wolbachia. The genes underlying CI, cifA and cifB, were recently identified in the eukaryotic association module of Wolbachia’s prophage WO. Here, we use transcriptomic and genomic approaches to address three important evolutionary facets of the cif genes. First, we assess whether or not cifA and cifB comprise a classic toxin–antitoxin operon in wMel and show that the two genes exhibit striking, transcriptional differences across host development. They can produce a bicistronic message despite a predicted hairpin termination element in their intergenic region. Second, cifA and cifB strongly coevolve across the diversity of phage WO. Third, we provide new domain and functional predictions across homologs within Wolbachia, and show that amino acid sequences vary substantially across the genus. Finally, we investigate conservation of cifA and cifB and find frequent degradation and loss of the genes in strains that no longer induce CI. Taken together, we demonstrate that cifA and cifB exhibit complex transcriptional regulation in wMel, provide functional annotations that broaden the potential mechanisms of CI induction, and report recurrent erosion of cifA and cifB in non-CI strains, thus expanding our understanding of the most widespread form of reproductive parasitism.
A Wolbachia deubiquitylating enzyme induces cytoplasmic incompatibility
5932Beckmann, J. F., J. A. Ronau and M. Hochstrasser, Nature Microbiology, 2:17007. 2017-03-01 17:45:04.
Wolbachia are obligate intracellular bacteria1 that infect arthropods, including approximately two-thirds of insect species2. Wolbachia manipulate insect reproduction by enhancing their inheritance through the female germline. The most common alteration is cytoplasmic incompatibility (CI)3–5, where eggs from uninfected females fail to develop when fertilized by sperm from Wolbachia-infected males. By contrast, if female and male partners are both infected, embryos are viable. CI is a gene-drive mechanism impacting population structure6 and causing reproductive isolation7, but its molecular mechanism has remained unknown. We show that a Wolbachia deubiquitylating enzyme (DUB) induces CI. The CI-inducing DUB, CidB, cleaves ubiquitin from substrates and is encoded in a two-gene operon, and the other protein, CidA, binds CidB. Binding is strongest between cognate partners in cidA-cidB homologues. In transgenic Drosophila, the cidA-cidB operon mimics CI when sperm introduce it into eggs, and a catalytically inactive DUB does not induce sterility. Toxicity is recapitulated in yeast by CidB alone; this requires DUB activity but is rescued by coexpressed CidA. A paralogous operon involves a putative nuclease (CinB) rather than a DUB. Analogous binding, toxicity and rescue in yeast were observed. These results identify a CI mechanism involving interacting proteins that are secreted into germline cells by Wolbachia, and suggest new methods for insect control.
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.
Selfish genes in mosquitos
6274C. F. Curtis, Nature, 357:450. 1992-06-11 16:47:56.
Hurst and coUeagues1.2 state that "within any population of [the mosquito] Culex pipiens there are two sorts of individual, those that bear/harbour Wolbachia [bacteria] and those that do not". But, according to Yen and Barr3, all wild-type C. pipiens appropriately examined before 1973 were found to carry Wolbachia, It was only when Yen and Barr produced uninfected individuals artificially by tetracycline treatment that it was found that matings of uninfected females x infected males were sterile, whereas all matings by uninfected males were fertile.
Population replacement in Culex-fatigans by means of cytoplasmic incompatibility .2. Field cage experiments with overlapping generations
6276C. F. Curtis, Bulletin of the World Health Organization, 53:107-119. 1976-01-08 16:51:11.
Three experiments were carried out in field cages to test the principle of " transport" of a desirable gene or chromosome into a wild Culex fatigans population as a result of the sterility in cross-matings associated with cytoplasmic incompatibility. Cycling populations of Delhi origin were established in the cages and daily releases were made of the IS31B strain, which has Paris cytoplasm and carries a male-linked translocation. It was shown that, if sufficient releases were made to establish a majority of the Paris cytoplasmic type, complete replacement by this cytoplasmic type subsequently occurred. However, as a result of partial compatibility of males of the Delhi population with Paris females, " recombinant " males with Paris cytoplasm and no translocation were produced. In an experiment in which a continuous low rate of " immigration " of a strain of Delhi origin was simulated, a gradual increase of the Paris cytoplasm non-translocated type occurred, and renewed IS31B releases were necessary after 5 months to restore the predominance of this type. The results are compared with computer predictions and discussed in relation to the transport of genes for filaria refractoriness or chromosome translocations into wild populations.
Population replacement in Culex fatigens by means of cytoplasmic incompatibility. Laboratory experiments with non-overlapping generations
6291C. F. Curtis and T. Adak, Bulletin of the World Health Organization, 51:249-255. 1974-01-08 19:45:42.
Bidirectional cytoplasmic incompatibility in the Culex pipiens complex appears to provide a mechanism for the replacement of a wild population by a strain refractory to filaria or a strain made partly sterile by a translocation. As a preliminary test of the feasibility of the replacement process, various ratios of strains with the cytoplasm of either Delhi or Paris, which are bidirectionally incompatible, were tested in laboratory cages. Where one strain was marked with the ruby-eye gene, this strain always declined in frequency in the next generation. In experiments in which the Paris strain was marked with a male-linked translocation complex, after 2-4 generations of breeding there was complete elimination of either the Paris or the Delhi type depending, as expected, on the relative frequencies of the two types with which the population began. In one experiment a type with Paris cytoplasm devoid of the translocation was found. This type increased in frequency in succeeding generations. The possible causes of origin of this type and its relevance to the practical use of the replacement principle are discussed.
Eradication of Culex pipiens fatigans through cytoplasmic incompatibility.
6278H. Laven, Nature, 216:383. 1967-10-28 16:57:49.
Culex pipiens fatigans is the chief vector of filariasis in south-east Asia. Urbanization has often caused the numbers of this mosquito-and with it the danger of filariasis infection-to increase alarmingly. The natural vigour, tolerance and fast development of resistance to insecticides of this mosquito necessitate the development of other control methods, and cytoplasmic incompatibility2 seems to be an ideal means.
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