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The intention-to-treat (ITT) analysis of the Applying Wolbachia to Eliminate Dengue (AWED) trial estimated a protective efficacy of 77.1% for participants resident in areas randomised to receive releases of wMel-infected Aedes aegypti mosquitoes, an emerging dengue preventive intervention. The limiting assumptions of ITT analyses in cluster randomised trials and the mobility of mosquitoes and humans across cluster boundaries indicate the primary analysis is likely to underestimate the full public health benefit. Using spatiotemporally-resolved data on the distribution of Wolbachia mosquitoes and on the mobility of AWED participants (n = 6306), we perform complier-restricted and per-protocol re-examinations of the efficacy of the Wolbachia intervention. Increased intervention efficacy was estimated in all analyses by the refined exposure measures. The complier-restricted analysis returned an estimated efficacy of 80.7% (95% CI 65.9, 89.0) and the per-protocol analysis estimated 82.7% (71.7, 88.4) efficacy when comparing participants with an estimated wMel exposure of ≥ 80% compared to those with

The Asian Citrus Psyllid (ACP), Diaphorina citri, is a vector of the pathological bacterium Candidatus Liberibacter asiaticus (CLas), which causes the most devastating disease to the citrus industry worldwide, known as greening or huanglongbing (HLB). Earlier field tests with an acetic acid-based lure in greening-free, Valencia citrus orange groves in California showed promising results. The same type of lures tested in São Paulo, Brazil, showed unsettling results. During the unsuccessful trials, we noticed a relatively large proportion of females in the field, ultimately leading us to test field-collected males and females for Wolbachia and CLas. The results showed high rates of Wolbachia and CLas infection in field populations. We then compared the olfactory responses of laboratory-raised, CLas-free, and CLas-infected males to acetic acid. As previously reported, CLas-uninfected males responded to acetic acid at 1 µg. Surprisingly, CLas-infected males required 50 × higher doses of the putative sex pheromone, thus explaining the failure to capture CLas-infected males in the field. CLas infection was also manifested in electrophysiological responses. Electroantennogram responses from CLas-infected ACP males were significantly higher than those obtained with uninfected males. To the best of our knowledge, this is the first report of a pathogen infection affecting a vectors response to a sex attractant.

Background: Five species of the Phortica genus (Diptera: Drosophilidae) are known in Europe and the Middle East. Among these, Phortica variegata and Phortica okadai are better known for their role as vectors of the zoonotic eyeworm Thelazia callipaeda. Other species, such as Phortica semivirgo and Phortica oldenbergi, have been studied less. Given the paucity of data about these Phortica spp. vectors, we explored the population dynamics and ecology of Phortica spp. in an area highly endemic for T. callipeada (Manziana, Rome, Central Italy). Methods: Phortica spp. flies were collected over a 3-year period (2018–2020) during their active season (April–October) with a sweep net while hovering around fermenting fruits or a human operator acting as baits. Collected flies were morphologically identified and tested for a T. callipeada infection and for the presence of Wolbachia, by polymerase chain reaction (PCR). Population dynamics of species collected was associated to environmental drivers through generalized additive models. Results: Of the 5564 flies collected, 90.8% were P. variegata, 9.1% were P. oldenbergi, 0.05% were P. semivirgo, and one specimen was P. okadai. Only P. variegata scored molecularly infected with T. callipeada throughout the 3-year sampling period (1.8%). Phortica oldenbergi, observed consistently during the entire sampling period, exhibited a marked preference for fruit traps, contrasting with the lachryphagous activity of P. variegata. Analysis of environmental drivers of P. oldenbergi and P. variegata population dynamics indicated temperature, wind speed, and pressure as significant factors. In addition, Wolbachia pipientis endosymbiont was detected in P. oldenbergi and P. okadai. Conclusions: For the first time, this study analysed several ecological aspects of Phortica species coexisting in a T. callipeada endemic area, highlighting different behaviors in the same environment and their vectorial role. Notably, this is also the first report of the presence of P. oldenbergi in Italy and P. okadai in Europe, underscoring the importance of extensive sampling for detecting potential vectors and alien species with direct implications for vector-borne disease epidemiology. [ABSTRACT FROM AUTHOR]

Wolbachia pipientis (= Wolbachia) has promise as a tool to suppress virus transmission by Aedes aegypti mosquitoes. However, Wolbachia can have variable effects on mosquito-borne viruses. This variation remains poorly characterized, yet the multimodal effects of Wolbachia on diverse pathogens could have important implications for public health. Here, we examine the effects of transient somatic infection with two strains of Wolbachia (wAlbB and wMel) on the alphaviruses Sindbis virus (SINV), O'nyong-nyong virus (ONNV), and Mayaro virus (MAYV) in Ae. aegypti. We found variable effects of Wolbachia including enhancement and suppression of viral infections, with some effects depending on Wolbachia strain. Both wAlbB- and wMel-infected mosquitoes showed enhancement of SINV infection rates one week post-infection, with wAlbB-infected mosquitoes also having higher viral titers than controls. Infection rates with ONNV were low across all treatments and no significant effects of Wolbachia were observed. The effects of Wolbachia on MAYV infections were strikingly strain-specific; wMel strongly blocked MAYV infections and suppressed viral titers, while wAlbB had more modest effects. The variable effects of Wolbachia on vector competence underscore the importance of further research into how this bacterium impacts the virome of wild mosquitoes including the emergent human pathogens they transmit. Author summary: In recent years, wild populations of Aedes aegypti mosquitoes in over a dozen countries have been deliberately infected with Wolbachia pipientis ("Wolbachia"); an intracellular bacterium that, in some circumstances, helps to curb the spread of mosquito-brone pathogens including dengue virus. But how does Wolbachia affect the ability of mosquitoes to become infected with and spread the many different viruses they encounter in nature? Here, we use transient somatic infections in Aedes aegypti to characterize the effects of Wolbachia on three different alphaviruses that cause illness in humans: Sindbis virus, O'nyong-nyong virus, and Mayaro virus. We find that transient Wolbachia infections have variable effects on these different pathogens, ranging from significant suppression of Mayaro virus to significant enhancement of Sindbis virus. Our research has important implications for the design of vector control strategies, and suggests further research is needed to understand how Wolbachia shapes the replication and transmission of diverse viruses in mosquitoes. [ABSTRACT FROM AUTHOR]

A good knowledge of the evolutionary history of organisms and an accurate taxonomic framework are keys for efficient biodiversity conservation measures. This is particularly relevant for European butterflies, which are often used as bioindicator organisms and umbrella species for conservation.The Holarctic butterfly genus Agriades includes seven European arctic‐alpine species of which several have a debated taxonomic status. They are localized and allopatric, and at least two of them are of high conservation concern, namely A. aquilo (in Europe, restricted to northern Scandinavia) and A. zullichi (endemic to southern Spain). Despite high conservation value and taxonomic uncertainty, molecular studies on these taxa are limited to DNA barcodes.We investigated the evolutionary history and species delimitation of West Palearctic Agriades using double‐digest RAD sequencing (ddRADseq).Various analyses using genomic data suggested the presence of one species in the glandon species group and one or two in the pyrenaicus species group. The differentiation within the former group (i.e., glandon, aquilo, zullichi) appears to be relatively recent, although the main allopatric populations represent evolutionary significant units (ESUs). Notable geographic structure was found within the pyrenaicus species group, with four allopatric ESUs detected.Wolbachia screening highlighted the presence of several allele combinations, some associated with particular Agriades populations.Genomic data revealed the evolutionary trajectory of European Agriades and a tendency for taxonomic oversplitting within this genus. These findings aid conservation by providing genetic background for region or population prioritization (including ESUs), and for the management of potential population reinforcement or reintroductions. [ABSTRACT FROM AUTHOR]

[Display omitted] • Tyrophagus putrescentiae (mould mite) is ubiquitous; its DNA contaminates sequences of other organisms in public databases. • Symbiotic bacteria are diverse, and aid the mite's success as a trophic generalist. • Tyrophagus putrescentiae is host to a Gram-negative bacterium, Wolbachia (supergroup Q) • Wolbachia supergroup Q is a basal, mite-specific lineage. • No horizontal gene transfer events from bacteria or fungi were found. Tyrophagus putrescentiae (mould mite) is a global, microscopic trophic generalist that commonly occurs in various human-created habitats, causing allergies and damaging stored food. Its ubiquity and extraordinary ability to penetrate research samples or cultures through air currents or by active walking through tights spaces (such as treads of screw caps) may lead to sample contamination and introduction of its DNA to research materials in the laboratory. This prompts a thorough investigation into potential sequence contamination in public genomic databases. The trophic success of T. putrescentiae is primarily attributed to the symbiotic bacteria housed in specialized internal mite structures, facilitating adaptation to varied nutritional niches. However, recent work suggests that horizontal transfer of bacterial/fungal genes related to nutritional functionality may also contribute to the mite's trophic versatility. This aspect requires independent confirmation. Additionally, T. putrescentiae harbors an uncharacterized and genetically divergent bacterium, Wolbachia , displaying blocking and microbiome-modifying effects. The phylogenomic position and supergroup assignment of this bacterium are unknown. Here, we sequenced and assembled the T. putrescentiae genome, analyzed its microbiome, and performed detailed phylogenomic analyses of the mite-specific Wolbachia. We show that T. putrescentiae DNA is a substantial source of contamination of research samples. Its DNA may inadvertently be co-extracted with the DNA of the target organism, eventually leading to sequence contamination in public databases. We identified a diversity of bacterial species associated with T. putrescentiae , including those capable of rapidly developing antibiotic resistance, such as Escherichia coli. Despite the presence of diverse bacterial communities in T. putrescentiae , we did not detect any recent horizontal gene transfers in this mite species and/or in astigmatid (domestic) mites in general. Our phylogenomic analysis of Wolbachia recovered a basal, mite-specific lineage (supergroup Q) represented by two Wolbachia spp. from the mould mite and a gall-inducing plant mite. Fluorescence in situ hybridization confirmed the presence of Wolbachia inside the mould mite. The discovery of an early derivative Wolbachia lineage (supergroup Q) in two phylogenetically unrelated and ecologically dissimilar mites suggests that this endosymbiotic bacterial lineage formed a long-term association with mites. This finding provides a unique insight into the early evolution and host associations of Wolbachia. Further discoveries of Wolbachia diversity in acariform mites are anticipated. [ABSTRACT FROM AUTHOR]

Bioinformatics sequence data mining can reveal hidden microbial symbionts that might normally be filtered and removed as contaminants. Data mining can be helpful to detect Wolbachia, a widespread bacterial endosymbiont in insects and filarial nematodes whose distribution in plant-parasitic nematodes (PPNs) remains underexplored. To date, Wolbachia has only been reported a few PPNs, yet nematode-infecting Wolbachia may have been widespread in the evolutionary history of the phylum based on evidence of horizontal gene transfers, suggesting there may be undiscovered Wolbachia infections in PPNs. The goal of this study was to more broadly sample PPN Wolbachia strains in tylenchid nematodes to enable further comparative genomic analyses that may reveal Wolbachia's role and identify targets for biocontrol. Published whole-genome shotgun assemblies and their raw sequence data from 33 Meloidogyne spp. assemblies, seven Globodera spp. assemblies, and seven Heterodera spp. assemblies were analyzed to look for Wolbachia. No Wolbachia was found in Meloidogyne spp. and Globodera spp., but among seven genome assemblies for Heterodera spp., an H. schachtii assembly from the Netherlands was found to have a large Wolbachia-like sequence that, when re-assembled from reads, formed a complete, circular genome. Detailed analyses comparing read coverage, GC content, pseudogenes, and phylogenomic patterns clearly demonstrated that the H. schachtii Wolbachia represented a novel strain (hereafter, denoted wHet). Phylogenomic tree construction with PhyloBayes showed wHet was most closely related to another PPN Wolbachia, wTex, while 16S rRNA gene analysis showed it clustered with other Heterodera Wolbachia assembled from sequence databases. Pseudogenes in wHet suggested relatedness to the PPN clade, as did the lack of significantly enriched GO terms compared to PPN Wolbachia strains. It remains unclear whether the lack of Wolbachia in other published H. schachtii isolates represents the true absence of the endosymbiont from some hosts. [ABSTRACT FROM AUTHOR]

Background: Mosquitoes serve as vectors for numerous pathogens, posing significant health risks to humans and animals. Understanding the complex interactions within mosquito microbiota is crucial for deciphering vector-pathogen dynamics and developing effective disease management strategies. Here, we investigated the nested patterns of Wolbachia endosymbionts and Escherichia-Shigella within the microbiota of laboratory-reared Culex pipiens f. molestus and Culex quinquefasciatus mosquitoes. We hypothesized that Wolbachia would exhibit a structured pattern reflective of its co-evolved relationship with both mosquito species, while Escherichia-Shigella would display a more dynamic pattern influenced by environmental factors. Results: Our analysis revealed different microbial compositions between the two mosquito species, although some microorganisms were common to both. Network analysis revealed distinct community structures and interaction patterns for these bacteria in the microbiota of each mosquito species. Escherichia-Shigella appeared prominently within major network modules in both mosquito species, particularly in module P4 of Cx. pipiens f. molestus, interacting with 93 nodes, and in module Q3 of Cx. quinquefasciatus, interacting with 161 nodes, sharing 55 nodes across both species. On the other hand, Wolbachia appeared in disparate modules: module P3 in Cx. pipiens f. molestus and a distinct module with a single additional taxon in Cx. quinquefasciatus, showing species-specific interactions and no shared taxa. Through computer simulations, we evaluated how the removal of Wolbachia or Escherichia-Shigella affects network robustness. In Cx. pipiens f. molestus, removal of Wolbachia led to a decrease in network connectivity, while Escherichia-Shigella removal had a minimal impact. Conversely, in Cx. quinquefasciatus, removal of Escherichia-Shigella resulted in decreased network stability, whereas Wolbachia removal had minimal effect. Conclusions: Contrary to our hypothesis, the findings indicate that Wolbachia displays a more dynamic pattern of associations within the microbiota of Culex pipiens f. molestus and Culex quinquefasciatus mosquitoes, than Escherichia-Shigella. The differential effects on network robustness upon Wolbachia or Escherichia-Shigella removal suggest that these bacteria play distinct roles in maintaining community stability within the microbiota of the two mosquito species. [ABSTRACT FROM AUTHOR]

AbstractIn this paper, Wolbachia infection in a bisexual and diploid population with a fixed cytoplasmic incompatibility rate w and maternal transmission rate d is studied as an evolution algebra. As the cytoplasmic incompatibility (CI) of the population causes deaths in the offspring, the evolution algebra of this model is not baric, and is a dibaric algebra if and only if the cytoplasmic incompatibility rate w is 1 and d=1 . The idempotent elements are given in terms of d and w. Moreover, this algebra has no absolute nilpotent elements when CI expression w≠1 . [ABSTRACT FROM AUTHOR]

Although new data and methodologies are gaining prominence in the field of taxonomy, the vast majority of terrestrial isopod species are still described solely based on morphology. The genus Benthana was recently revised and currently comprises 30 species found in South America. However, the broad geographic distribution of some species, coupled with the presence of few diagnostic characters, raises doubts about the validity of the described species. This study aimed to test the applicability of DNA Barcoding to assess the congruence between morphological and molecular identification of Benthana species. We analyzed 72 individuals belonging to 13 species from the scientific collection of the UFRGS Crustacean Laboratory. We obtained 24 sequences from ten Benthana species. The molecular phylogenies demonstrated that Benthana forms a paraphyletic group. Even at the species level, paraphyletic taxa were observed. Species delimitation methods revealed the presence of eight molecular entities within Benthana. Regarding genetic distances, we observed low intra-specific distances and significantly high inter-specific distances. Our results show that molecular analysis does not support the current taxonomy of Benthana. Therefore, a reassessment of the genus systematics based on integrative taxonomy is warranted. [ABSTRACT FROM AUTHOR]

The impact of selection on host immune function genes has been widely documented. However, it remains essentially unknown how mutation influences the quantitative immune traits that selection acts on. Applying a classical mutation accumulation (MA) experimental design in Drosophila serrata , we found the mutational variation in susceptibility (median time of death, LT50) to Drosophila C virus (DCV) was of similar magnitude to that reported for intrinsic survival traits. Mean LT50 did not change as mutations accumulated, suggesting no directional bias in mutational effects. Maintenance of genetic variance in immune function is hypothesized to be influenced by pleiotropic effects on immunity and other traits that contribute to fitness. To investigate this, we assayed female reproductive output for a subset of MA lines with relatively long or short survival times under DCV infection. Longer survival time tended to be associated with lower reproductive output, suggesting that mutations affecting susceptibility to DCV had pleiotropic effects on investment in reproductive fitness. Further studies are needed to uncover the general patterns of mutational effect on immune responses and other fitness traits, and to determine how selection might typically act on new mutations via their direct and pleiotropic effects. [ABSTRACT FROM AUTHOR]

Simple Summary: Maternally transmitted Wolbachia is one of the most common endosymbionts in many arthropods, influencing various aspects of host physiology, reproduction, and fitness. While recent studies on Wolbachia-Drosophila systems have revealed its influence on host thermal choice behavior, its broader impact on host thermal preference (Tp) in non-model insects remains poorly understood. The polyphagous leaf-miner fly, Liriomyza huidobrensis (Blanchard), one of the most notorious pests of vegetable and flowers globally, harbors a range of bacterial symbionts, with Wolbachia being especially prevalent. This study aims to explore the effects of Wolbachia on the thermal preference of the invasive leaf-miner L. huidobrensis. Understanding the potential roles of Wolbachia in host thermal behavior is crucial for elucidating the co-evolution of invasive species and their endosymbiont Wolbachia in the context of global climate change and temperature extremes, and may offer insight for the development of pest control strategies utilizing Wolbachia. Heritable endosymbiont Wolbachia is prevalent among arthropods, serving multiple functions for their hosts. However, the role of Wolbachia in mediating thermal preference selection remains largely unexplored. In this study, we utilized a custom-built thermal gradient to evaluate the thermal preference (Tp) of 1367 individuals of the invasive leaf-miner Liriomyza huidobrensis with or without Wolbachia wLhui from Yunnan and Xinjiang populations. Under meticulously controlled conditions and with a vast sample size, we found no significant difference in the mean Tp between wLhui-infected and uninfected leaf miners from either population when host age and sex were not considered. Furthermore, generalized linear model (GLM) analysis revealed no significant correlation between average Tp and age, sex, or Wolbachia infection, nor interactions among these factors, except in the Xinjiang population, where Tp was strongly associated with host age. Finally, we discuss the ecological implications of these findings and propose future research directions on Wolbachia-mediated host Tp in the leaf miner. Overall, our findings do not provide evidence that Wolbachia significantly affects the thermal preference of L. huidobrensis. Further studies across different systems are needed to investigate the complex interactions between Wolbachia and insect thermal behavior. [ABSTRACT FROM AUTHOR]

Background Matings between male Aedes aegypti mosquitoes infected with w AlbB strain of Wolbachia and wildtype females yield non-viable eggs. We evaluated the efficacy of releasing w AlbB-infected Ae. aegypti male mosquitoes to suppress dengue. Methods We specified the protocol of a two-arm cluster-randomized test-negative controlled trial (cRCT) and emulated it using a nationally representative test-negative/positive database of individuals reporting for febrile illness to any public hospital, general practitioner or polyclinic. We retrospectively built a cohort of individuals who reside in Wolbachia locations vs a comparator control group who do not reside in Wolbachia locations, using a nationally representative database of all individuals whom report for febrile illness and were tested for dengue at the Environmental Health Institute/hospital laboratories/commercial diagnostic laboratories, through general practitioner clinic, polyclinic or public/private hospital from epidemiological week (EW) 1 2019 to EW26 2022. We emulated a constrained randomization protocol used in cRCTs to balance dengue risk between intervention and control arms in the pre-intervention period. We used the inverse probability weighting approach to further balance the intervention and control groups using a battery of algorithmically selected sociodemographic, environmental and anthropogenic variables. Intention-to-treat analyses were conducted to estimate the risk reduction of dengue given Wolbachia exposure. Results Intention-to-treat analyses revealed that, compared with controls, Wolbachia releases for 3, 6 and ≥12 months was associated to 47% (95% confidence interval: 25–69%), 44% (33–77%) and 61% (38–78%) protective efficacy against dengue, respectively. When exposed to ≥12 months of Wolbachia releases, protective efficacies ranged from 49% (13–72%) to 77% (60–94%) across years. The proportion of virologically confirmed dengue cases was lower overall in the intervention arm. Protective efficacies were found across all years, age and sex subgroups, with higher durations of Wolbachia exposure associated to greater risk reductions of dengue. Conclusion Results demonstrated that Wolbachia -mediated sterility can strengthen dengue control in tropical cities, where dengue burden is the greatest. [ABSTRACT FROM AUTHOR]

Wolbachia are common heritable endosymbionts that influence many aspects of ecology and evolution in various insects, yet Wolbachia‐mediated intracellular metabolic responses to temperature stress have been largely overlooked. Here, we introduced the Wolbachia strain wLhui from the invasive Liriomyza huidobrensis (Blanchard) into a Drosophila Schneider 2 cell line (S2) and investigated the metabolite profile of wLhui‐infected (S2_wLhui) and uninfected cell lines (S2_wu) under short‐term exposure to either high (37°C), moderate (27°C), or low (7 and 17°C) temperatures. We find that Wolbachia infection, temperature stress, and their interactions significantly affect cellular metabolic profiles. Most significantly, when comparing the changes in metabolites between S2_wLhui and S2_wu, glycerophospholipids, amino acids, and fatty acids associated with metabolic pathways, microbial metabolism in diverse environments, and other pathways were significantly accumulated at either low or high temperatures. Our findings suggest Wolbachia‐induced cellular physiological responses to short‐term temperature stress, which may in turn affect the fitness and adaptive ability of its host as an invasive species. [ABSTRACT FROM AUTHOR]

There is increasing evidence that host–parasitoid interactions can have a pronounced impact on the microbiome of host insects, but it is unclear to what extent this is caused by the host and/or parasitoid. Here, we compared the internal and external microbiome of caterpillars of Pieris brassicae and Pieris rapae parasitized by Cotesia glomerata or Cotesia rubecula with nonparasitized caterpillars. Additionally, we investigated the internal and external microbiome of the parasitoid larvae. Both internal and external bacterial densities were significantly higher for P. brassicae than P. rapae , while no differences were found between parasitized and nonparasitized caterpillars. In contrast, parasitism significantly affected the composition of the internal and external microbiome of the caterpillars and the parasitoid larvae, but the effects were dependent on the host and parasitoid species. Irrespective of host species, a Wolbachia species was exclusively found inside caterpillars parasitized by C. glomerata , as well as in the corresponding developing parasitoid larvae. Similarly, a Nosema species was abundantly present inside parasitized caterpillars and the parasitoid larvae, but this was independent of the host and the parasitoid species. We conclude that parasitism has pronounced effects on host microbiomes, but the effects depend on both the host and parasitoid species. [ABSTRACT FROM AUTHOR]

The increasing spread of mosquito vectors has made mosquito-borne arboviral diseases a global threat to public health, leading to the urgent need for effective population control methods. Strategies based in the intracellular bacterium Wolbachia Hertig, 1936 are considered environmentally friendly, safe for humans, and potentially cost-effective for controlling arboviral diseases. To minimize undesirable side effects, it is relevant to assess whether Wolbachia is present in the area and understand the diversity associated with native infections before implementing these strategies. With this purpose, we investigated Wolbachia infection status, diversity, and prevalence in populations of Aedes albifasciatus (Macquart, 1838), Aedes fluviatilis (Lutz, 1904), and hybrids of the Culex pipiens (Linnaeus, 1758) complex from Argentina. Aedes albifasciatus and C. pipiens complex samples were collected in the province of Buenos Aires, and A. fluviatilis in the province of Misiones. Aedes albifasciatus was found to be uninfected, while infections with strains wFlu and wPip were detected in A. fluviatilis and hybrids of the C. pipiens complex, respectively. All strains were fixed or close to fixation and clustered within supergroup B. These finding provides valuable information on Wolbachia strains found in natural mosquito populations in Argentina that might be used in heterologous infections in the future or be considered when designing control strategies based on Wolbachia infection. [ABSTRACT FROM AUTHOR]

Dengue fever as a mosquito-borne disease caused by dengue virus is responsible for a substantial disease burden to the world. Wolbachia as an innovative technique has been approved to inhibit the replication of dengue viruses in mosquitoes or reduce the number of wild mosquitoes. We firstly establish a birth-pulse model with sex structures to depict the nonlinear dynamics of mosquito population and then to investigate how Wolbachia can suppress or replace wild mosquitoes. The existence and stability of periodic solutions of the system are proved by analyzing its stroboscopic map. Theoretical results show that under complete maternal transmission, Wolbachia will successfully establish and completely replace wild mosquitoes. Under incomplete maternal transmission, there may be two pairs of bistable periodic solutions, including the coexistence of replacement failure and partial replacement periodic solutions, or the coexistence of replacement failure and mosquito eradication periodic solutions. For these bistable situations, if the initial proportion of Wolbachia-carrying mosquitoes is greater than the critical threshold, then the strategy of partial replacement or mosquito eradication can be succeeded. Since the initial proportion of Wolbachia mosquitoes is not always greater enough in the field, we introduce mosquito releases into the birth-pulse model, and obtain the condition for the stability of complete replacement periodic solution. Finally, we give rich numerical simulations to show the multiple attractors of the systems, the comparison of three common release strategies and the effects of Wolbachia-induced parameters on control strategies. At last section, we summarize some key suggestions on the control of mosquitoes and dengue fever in practice. This work will be helpful for public health authorities in designing proper release strategies to control the spread of dengue fever. • Birth-pulse mosquito population models with sex structures are established. • Dynamics of the systems are investigated by analyzing their stroboscopic map. • Existence for two pairs of bistable periodic solutions are proved and simulated. • Effects of three release methods and control parameters on control strategies are discussed. [ABSTRACT FROM AUTHOR]

Background: Iran is one of the endemic areas of Dirofilaria immitis. The modified Knott test is the common diagnostic test based on detecting and identifying microfilaria in blood samples. Still, the results may be compromised by D. immitis occult infection (D. immitis infection without microfilariae). Wolbachia pipientis in a dog's blood is an endosymbiont of D. immitis and an indicator of dirofilariasis. Objectives: This study aimed to investigate the occult infection of D. immitis in stray dogs in Guilan, Qazvin, and Mazandaran provinces, Iran, based on W. pipientis DNA tracking. Methods: Blood samples (n=138) were collected, and the presence of W. pipientis was evaluated by amplifying a partial sequence of the FtsZ gene (267 bp). Results: The results showed that 17.4% of the samples were positive by the modified Knott method, while 47.8% were positive by the molecular methods. The results of high-resolution melting (HRM) real-time PCR showed that the melting temperature of cox1 gene amplicons of D. immitis and Acanthocheilonema reconditum were obtained at 76.67±0.03 °C and 81.38±0.05 °C, respectively as well as it was achieved 80.30±0.04 °C for FtsZ gene of W. pipientis. The results of the present study showed that it is necessary to use molecular methods to diagnose dirofilariasis occult infection accurately. Conclusion: The HRM real-time PCR analysis is a simple post-PCR step that exploits the thermal characteristics of the amplicons to detect the occult infection of dirofilariasis based on W. pipientis DNA tracking and identification. [ABSTRACT FROM AUTHOR]

The Enemy Release Hypothesis posits that invasive plants in secondary ranges are freed from phytophage pressure, yet local phytophages may adapt to these invasive species over time. This study investigated the dietary preferences of Altica oleracea in relation to three native (Chamaenerion angustifolium (L.) Scop., Epilobium montanum L., and E. hirsutum L.) and three invasive (E. adenocaulon Hausskn., Oenothera biennis L., Oe. rubricaulis Kleb.) plant species from the Onagraceae family. A neural network was employed to automate the calculation of leaf damage area for assessing the phytophagous insects' dietary preferences. A. oleracea demonstrated a pronounced preference for Chamaenerion angustifolium compared to other Onagraceae species, irrespective of their invasive status. Among imago, no significant preference was observed between native and invasive Epilobium species or between two Oenothera species. However, the larvae were more discerning and avoided Oe. biennis, rearing on which led to the least weight gain by larvae. The adult beetles' inability to differentiate between plants unsuitable for larvae could lead to an evolutionary trap. The prevalence of Wolbachia infection in the A. oleracea population may influence the further evolution of adaptability to invasive species and the formation of dietary preferences. [ABSTRACT FROM AUTHOR]

Wolbachia , a prevalent intracellular symbiotic bacterium in insects, plays a significant role in insect biology. Ectropis grisescens (Warren; Lepidoptera: Geometridae) is a devastating chewing pest distributed in tea plantations throughout China. However, it is unclear how Wolbachia titers affect the fitness and reproduction of E. grisescens. In this study, the impacts of 3 different infection lines, naturally Wolbachia- infected, Wolbachia- uninfected, and Wolbachia transinfected, regarding the life history traits of E. grisescens, were evaluated using the age-stage, 2-sex life table. Wolbachia infection shortened preadult duration and preoviposition periods and increased the fecundity, net reproductive rate, and finite rate of increase. Meanwhile, population projection indicated that E. grisescens population size with Wolbachia infection can increase faster than without. These results indicate that Wolbachia plays a regulatory role in the fitness of E. grisescens. It is also noted that the life history parameters of E. grisescens may positively correlate with Wolbachia titers. These findings could aid in pest management in tea gardens. [ABSTRACT FROM AUTHOR]

Wolbachia are the most spread bacterial endosymbionts in the world. These bacteria can manipulate host reproduction or block virus transmission in mosquitoes. For this reason, Wolbachia-based strategies for vector control are seriously considered or have already been applied in several countries around the world. In South America, Wolbachia have been studied in human pathogen vectors such as sand flies and mosquitoes. In French Guiana, the diversity and distribution of Wolbachia are not well known in mosquitoes. In this study, we screened for Wolbachia natural infection in mosquitoes in French Guiana by using 16S rRNA, Wolbachia surface protein (WSP), and multi-locus sequence typing (MLST) molecular assays. A total of 29 out of 44 (65.9%) mosquito species were positive for natural Wolbachia infection according to the PCR results, and two Wolbachia strains co-infected three specimens of Mansonia titillans. Then, we analyzed the phylogenetic relationships among the Wolbachia detected. All of the tested specimens of Aedes aegypti, the major dengue vector of French Guiana, were negative. These results regarding Wolbachia strain, distribution, and prevalence in mosquitoes from French Guiana highlight Wolbachia–mosquito associations and pave the way for a future Wolbachia-based strategy for vector control in this Amazonian territory. [ABSTRACT FROM AUTHOR]

BACKGROUND: Releasing large numbers of Aedes albopictus males, carrying the artificially introduced Wolbachia 'wPip' strain, results in a decrease in the reproductive capacity of wild females due to a phenomenon known as cytoplasmic incompatibility (CI). This vector control strategy is referred to as the incompatible insect technique (IIT). However, its widespread implementation faces various challenges, including the complexity of removing fertile females from the males intended for release. Here, we present the results of semi‐field experiments comparing the impact of minimal female co‐release on two IIT modes: unidirectional CI‐based (UnCI IIT) and bidirectional CI‐based (BiCI IIT), specifically targeting Ae. albopictus. RESULTS: The contamination of 'wPip' infected females (2%) during male releases significantly weakened the overall effectiveness of IIT, emphasizing the need for thorough sex separation. Specifically, with UnCI IIT, despite the low rate of co‐released females, there was a gradual rise in 'wPip' infection frequency, resulting in more compatible mating and subsequently higher rates of egg hatching. Conversely, this pattern was effectively mitigated in BiCI IIT owing to the reciprocal sterility between the wild‐type and the 'wPip' infected populations. CONCLUSION: Through an experimental approach, conducted in a semi‐field setting, we have contributed to advancing scientific understanding regarding the potential outcomes of implementing the IIT strategy in the absence of a complete sexing system. The results suggest that safety measures for mitigating the potential impacts of co‐released females can be tailored according to the specific type of IIT being utilized. © 2024 Society of Chemical Industry. [ABSTRACT FROM AUTHOR]

BACKGROUND: Wolbachia are widespread intracellular bacteria in insects that often have high rates of spread due to their impact on insect reproduction. These bacteria may also affect the mating behavior of their host with impacts on the fitness of host progeny. In this study, we investigated the impact of Wolbachia on a preference for mating with young or old males in the parasitoid wasp Habrobracon hebetor. RESULTS: Our results showed that uninfected females from a tetracycline‐treated line preferred to mate with young males, whereas Wolbachia‐infected females had no preference. Time to mating was relatively shorter in the infected lines. Regardless of Wolbachia infection status, progeny resulting from matings with young males showed higher fitness than those from crosses with old males, and infected females crossed with infected young males showed the highest performance. CONCLUSION: These results suggest an impact of Wolbachia on female mate preference and offspring fitness although it is unclear how this phenomenon increases Wolbachia transmission of infected wasps. © 2024 Society of Chemical Industry. [ABSTRACT FROM AUTHOR]

Background: Tsetse flies, the biological vectors of African trypanosomes, have established symbiotic associations with different bacteria. Their vector competence is suggested to be affected by bacterial endosymbionts. The current study provided the prevalence of three tsetse symbiotic bacteria and trypanosomes in Glossina species from Burkina Faso. Results: A total of 430 tsetse flies were captured using biconical traps in four different collection sites around Bobo-Dioulasso (Bama, Bana, Nasso, and Peni), and their guts were removed. Two hundred tsetse were randomly selected and their guts were screened by PCR for the presence of Sodalis glossinidius, Spiroplasma sp., Wolbachia and trypanosomes. Of the 200 tsetse, 196 (98.0%) were Glossina palpalis gambiensis and 4 (2.0%) Glossina tachinoides. The overall symbiont prevalence was 49.0%, 96.5%, and 45.0%, respectively for S. glossinidius, Spiroplasma and Wolbachia. Prevalence varied between sampling locations: S. glossinidius (54.7%, 38.5%, 31.6%, 70.8%); Spiroplasma (100%, 100%, 87.7%, 100%); and Wolbachia (43.4%, 38.5%, 38.6%, 70.8%), respectively in Bama, Bana, Nasso and Peni. Noteworthy, no G. tachnoides was infected by S. glossinidius and Wolbachia, but they were all infected by Spiroplasma sp. A total of 196 (98.0%) harbored at least one endosymbionts. Fifty-five (27.5%) carried single endosymbiont. Trypanosomes were found only in G. p. gambiensis, but not G. tachinoides. Trypanosomes were present in flies from all study locations with an overall prevalence of 29.5%. In Bama, Bana, Nasso, and Peni, the trypanosome infection rate was respectively 39.6%, 23.1%, 8.8%, and 37.5%. Remarkably, only Trypanosoma grayi was present. Of all trypanosome-infected flies, 55.9%, 98.3%, and 33.9% hosted S. glossinidius, Spiroplasma sp and Wolbachia, respectively. There was no association between Sodalis, Spiroplasma and trypanosome presence, but there was a negative association with Wolbachia presence. We reported 1.9 times likelihood of trypanosome absence when Wolbachia was present. Conclusion: This is the first survey reporting the presence of Trypanosoma grayi in tsetse from Burkina Faso. Tsetse from these localities were highly positive for symbiotic bacteria, more predominantly with Spiroplasma sp. Modifications of symbiotic interactions may pave way for disease control. [ABSTRACT FROM AUTHOR]

Dengue virus (DENV) is the most prevalent mosquito-borne Flavivirus that affects humans worldwide. Aedes albopictus, which is naturally infected with the bacteria Wolbachia, is considered to be a secondary vector of DENV. However, it was responsible for a recent DENV outbreak of unprecedented magnitude in Reunion Island, a French island in the South West Indian Ocean. Moreover, the distribution of the cases during this epidemic showed a spatially heterogeneous pattern across the island, leading to questions about the differential vector competence of mosquito populations from different geographic areas. The aim of this study was to gain a better understanding of the vector competence of the Ae. albopictus populations from Reunion Island for local DENV epidemic strains, while considering their infection by Wolbachia. Experimental infections were conducted using ten populations of Ae. albopictus sampled across Reunion Island and exposed to three DENV strains: one strain of DENV serotype 1 (DENV-1) and two strains of DENV serotype 2 (DENV-2). We analyzed three vector competence parameters including infection rate, dissemination efficiency and transmission efficiency, at different days post-exposition (dpe). We also assessed whether there was a correlation between the density of Wolbachia and viral load/vector competence parameters. Our results show that the Ae. albopictus populations tested were not able to transmit the two DENV-2 strains, while transmission efficiencies up to 40.79% were observed for the DENV-1 strain, probably due to difference in viral titres. Statistical analyses showed that the parameters mosquito population, generation, dpe and area of sampling significantly affect the transmission efficiencies of DENV-1. Although the density of Wolbachia varied according to mosquito population, no significant correlation was found between Wolbachia density and either viral load or vector competence parameters for DENV-1. Our results highlight the importance of using natural mosquito populations for a better understanding of transmission patterns of dengue. [ABSTRACT FROM AUTHOR]

Introduction: Symbiotic bacteria play key roles in a variety of important life processes of insects such as development, reproduction and environmental adaptation, and the elucidation of symbiont population structure and dynamics is crucial for revealing the underlying regulatory mechanisms. The marmalade hoverfly (Episyrphus balteatus) is not only a remarkable aphid predator, but also a worldwide pollinator second to honeybees. However, its symbiont composition and dynamics remain unclear. Methods: Herein, we investigate the symbiotic bacterial dynamics in marmalade hoverfly throughout whole life cycle, across two sexes, and in its prey Megoura crassicauda by 16S rRNA sequencing. Results: In general, the dominant phyla were Proteobacteria and Firmicutes, and the dominant genera were Serratia and Wolbachia. Serratia mainly existed in the larval stage of hoverfly with the highest relative abundance of 86.24% in the 1st instar larvae. Wolbachia was found in adults and eggs with the highest relative abundance of 62.80% in eggs. Significant difference in species diversity was observed between the adults feeding on pollen and larvae feeding on M. crassicauda, in which the dominant symbiotic bacteria were Asaia and Serratia, respectively. However, between two sexes, the symbionts exhibited high similarity in species composition. In addition, our results suggested that E. balteatus obtainded Serratia mainly through horizontal transmission by feeding on prey aphids, whereas it acquired Wolbachia mainly through intergeneration vertical transmission. Taken together, our study revealed the effects of development stages, diet types and genders of E. balteatus on symbionts, and explored transmission modes of dominant bacteria Serratia and Wolbachia. Discussion: Our findings lay a foundation for further studying the roles of symbiotic bacteria in E. balteatus life cycle, which will benefit for revealing the co-adaptation mechanisms of insects and symbiotic bacteria. [ABSTRACT FROM AUTHOR]

Introduction: Endosymbiotic bacteria in the genus Wolbachia have evolved numerous strategies for manipulating host reproduction in order to promote their own transmission. This includes the feminization of males into functional females, a well-studied phenotype in the isopod Armadillidium vulgare. Despite an early description of this phenotype in isopods and the development of an evolutionary model of host sex determination in the presence of Wolbachia, the underlying genetic mechanisms remain elusive. Methods: Here we present the first complete genomes of the three feminizing Wolbachia (wVulC, wVulP, and wVulM) known to date in A. vulgare. These genomes, belonging to Wolbachia B supergroup, contain a large number of mobile elements such as WO prophages with eukaryotic association modules. Taking advantage of these data and those of another Wolbachia-derived feminizing factor integrated into the host genome (f element), we used a comparative genomics approach to identify putative feminizing factors. Results: This strategy has enabled us to identify three prophage-associated genes secreted by the Type IV Secretion System: one ankyrin repeat domaincontaining protein, one helix-turn-helix transcriptional regulator and one hypothetical protein. In addition, a latrotoxin-related protein, associated with phage relic genes, was shared by all three genomes and the f element. Conclusion: These putative feminization-inducing proteins shared canonical interaction features with eukaryotic proteins. These results pave the way for further research into the underlying functional interactions. [ABSTRACT FROM AUTHOR]

The mosquito microbiome significantly influences vector competence, including in Aedes albopictus, a globally invasive vector. Describing the microbiome and Wolbachia strains of Ae. albopictus from different regions can guide area-specific control strategies. Mosquito samples from Spain and São Tomé were analyzed using 16S rRNA gene sequencing and metagenomic sequencing. Wolbachia infection patterns were observed by sex and population. Female mosquitoes were blood-fed, a factor considered in analyzing their microbiota. Results revealed a dominance of dual Wolbachia infections, strains A and B, in the microbiome of both populations of Ae. albopictus, especially among females. Both populations shared a core microbiome, although 5 and 9 other genera were only present in Spain and São Tomé populations, respectively. Genera like Pelomonas and Nevskia were identified for the first time in Aedes mosquitoes. This study is the first to describe the Ae. albopictus bacteriome in Spain and São Tomé, offering insights for the development of targeted mosquito control strategies. Understanding the specific microbiome composition can help in designing more effective interventions, such as microbiome manipulation and Wolbachia-based approaches, to reduce vector competence and transmission potential of these mosquitoes. [ABSTRACT FROM AUTHOR]

Simple Summary: Bemisia tabaci is as a major pest of vegetable crops in Cameroon, and several species have developed resistance against insecticides. Here, we investigated the frequency of infection by endosymbiont and the genetic diversity of whiteflies in Cameroon. Mitochondrial cytochrome oxidase I (mtCOI) markers and Kompetitive Allele Specific PCR (KASP) were used for the characterization. Overall, an analysis of the mtCOI sequences showed six mitotypes of Bemisia tabaci, and two distinct clades of Bemisia afer and Trialeurodes vaporariorum. Bemisia tabaci mitotypes identified included: Mediterranean (MED) on tomato, pepper, okra, and melon; and sub-Saharan Africa (SSA) groups and sub-groups (SG)–SSA1-SG1, SSA1-SG2, SSA1-SG5, SSA3, and SSA4 on cassava. The six mitotypes of cassava B. tabaci were split into three SNP haplogroups including sub-Saharan Africa–West Africa (SSA-WA), sub-Saharan East and Central Africa (SSA-ECA), and SSA4 by KASP genotyping. The endosymbionts identified infecting the whiteflies were Arsenophonus, Rickettsia, and Wolbachia. Bemisia tabaci (Gennadius) is as a major pest of vegetable crops in Cameroon. These sap-sucking insects are the main vector of many viruses infecting plants, and several cryptic species have developed resistance against insecticides. Nevertheless, there is very little information about whitefly species on vegetable crops and the endosymbionts that infect them in Cameroon. Here, we investigated the genetic diversity of whiteflies and their frequency of infection by endosymbionts in Cameroon. Ninety-two whitefly samples were collected and characterized using mitochondrial cytochrome oxidase I (mtCOI) markers and Kompetitive Allele Specific PCR (KASP). The analysis of mtCOI sequences of whiteflies indicated the presence of six cryptic species (mitotypes) of Bemisia tabaci, and two distinct clades of Bemisia afer and Trialeurodes vaporariorum. Bemisia tabaci mitotypes identified included: MED on tomato, pepper, okra, and melon; and SSA1-SG1, SSA1-SG2, SSA1-SG5, SSA3, and SSA4 on cassava. The MED mitotype predominated in all regions on the solanaceous crops, suggesting that MED is probably the main phytovirus vector in Cameroonian vegetable cropping systems. The more diverse cassava-colonizing B. tabaci were split into three haplogroups (SNP-based grouping) including SSA-WA, SSA4, and SSA-ECA using KASP genotyping. This is the first time that SSA-ECA has been reported in Cameroon. This haplogroup is predominant in regions currently affected by the severe cassava mosaic virus disease (CMD) and cassava brown streak virus disease (CBSD) pandemics. Three endosymbionts including Arsenophonus, Rickettsia, and Wolbachia were present in female whiteflies tested in this study with varying frequency. Arsenophonus, which has been shown to influence the adaptability of whiteflies, was more frequent in the MED mitotype (75%). Cardinium and Hamiltonella were absent in all whitefly samples. These findings add to the knowledge on the diversity of whiteflies and their associated endosymbionts, which, when combined, influence virus epidemics and responses to whitefly control measures, especially insecticides. [ABSTRACT FROM AUTHOR]

Simple Summary: The mosquito species Culex pipiens, has two ecotypes, pipiens and molestus. The two forms differ in mating behavior and how they manage overwintering, with the molestus form thriving in closed indoor surroundings but without an ability to manage cold winter conditions, and the pipiens form that requiring open spaces for mating and going into diapause during winter. Both ecotypes are important vectors for mosquito-borne viruses. Cx. pipiens mosquitoes often carry endosymbiotic Wolbachia bacteria that are transmitted from the mother to the egg, but this varies frequency of occurrence. The Wolbachia endosymbiont affects the reproductive success of mosquitoes so that only eggs infected with a compatible Wolbachia can be fertilized by an infected male. In artificially infected Aedes mosquitoes, Wolbachia affects whether the mosquito can transmit a virus though bites. We studied how Cx. pipiens mosquitoes from Sweden carried Wolbachia and compared them to mosquitoes from other European countries, and found that all tested Cx. pipiens mosquitoes carried Wolbachia. We characterized the Wolbachia from these mosquitoes and found that Cx. pipiens of the two ecotypes carry different strains of Wolbachia in Sweden and Norway but not in sampled mosquitoes from Italy or England. This highlights the differences between the two ecotypes in northern Europe. The mosquito Culex pipiens occurs in two ecotypes differing in their mating and overwintering behavior: pipiens mate in open environments and diapause, and molestus also mate in small spaces and is active throughout the year. Cx. pipiens carry Wolbachia endosymbionts of the wPip strain, but the frequency of infection differs between studied populations. Wolbachia infection affects the host reproductive success through cytoplasmic incompatibility. wPip Wolbachia is divided into five types, wPip I–V. The type of wPip carried varies among Cx. pipiens populations. In northern European locations different wPip types are found in the two ecotypes, whereas in southern locations, they often carry the same type, indicating differences in hybridization between ecotypes. In this study, Cx. pipiens specimens of both ecotypes were collected from Sweden and compared to specimens from Norway, England, Italy, and the Netherlands, as well as Cx. quinquefasciatus from Mali and Thailand. The abundance varied, but all specimens were infected by Wolbachia, while the tested specimens of other mosquito species were often uninfected. The wPip strains were determined through the sequence analysis of Wolbachia genes ank2 and pk1, showing that Cx. pipiens ecotypes in Scandinavia carry different wPip strains. The observed differences in wPip strains indicate that hybridization is not frequent and may contribute to barriers against hybridization of the ecotypes in Sweden and Norway. [ABSTRACT FROM AUTHOR]

Moringa oleifera Lam. (moringa) is a plant native to India, used as a nutritional and medicinal supplement in many cultures around the world. Moringa has been linked to maintaining metabolic homeostasis and is often marketed as a weight loss supplement and a potential remedy for diseases such as diabetes. Here, we investigate how moringa, a 'superfood' with predicted protective effects against chronic diseases such as diabetes, influences the nutritional physiology and microbiome composition of the fruit fly Drosophila melanogaster. We administered moringa as a dietary supplement to Drosophila, and quantified key nutritional indices: glucose, triacylglyceride, and protein levels, and fly weight. We showed that dietary moringa supplementation significantly reduced fly glucose levels by up to ~30% and resulted in substantial restructuring of Drosophila microbiota composition, altering both gut and intracellular bacterial populations. The effect of moringa on fly glucose levels is specific because other nutritional indices, namely, triacylglyceride and protein levels and fly weight, were not significantly affected by dietary moringa supplementation. This study highlights the importance of moringa as a modulator of host glucose metabolism. [ABSTRACT FROM AUTHOR]

Once infected with parthenogenesis inducing (PI) Wolbachia, the reproductive mode of Trichogramma pretiosum would change from bisexual to thelytokous parthenogenesis; however, male offspring would also appear after maternal females' continuous massive oviposition. Whether there was a relationship between Wolbachia titer and sex determination in T. pretiosum was unclear. In this study, we compared the sex ratio of the offspring of parthenogenetic and bisexual restored lines of T. pretiosum, and measured the Wolbachia titer in parthenogenetic line of T. pretiosum before and after oviposition, in different body parts, as well as at different developmental stages. Results indicated that all the offspring produced by parthenogenetic line females on the 1st day of egg-laying were females, and male offspring were gradually appearing with the increase of females' oviposition age; but the male offspring began to appear on the 1st egg-laying day of bisexual restored line. After oviposition, Wolbachia titer in maternal female of parthenogenetic line was maintained at a lower level compared to those of pre-oviposition females. QRT-PCR results showed that Wolbachia in male offspring was significantly lower than that in female ones; and Wolbachia could be detected in all body parts of female parasitoids, and the titer was highest in abdomen. At different developmental stages, relative Wolbachia titer was higher in eggs and adults than other stages, and the Wolbachia titer in ovaries post-oviposition was significantly lower than that of pre-oviposition. In conclusion, Wolbachia titer is closely related to the sex determination of T. pretiosum. [ABSTRACT FROM AUTHOR]

Transposable elements make up substantial proportions of eukaryotic genomes and many are thought to be remnants of ancient viral infections. Current research has begun to highlight the role transposable elements can play in the immune system response to infections. However, most of our knowledge about transposable element expression during infection is limited by the specific host and pathogen factors from each study, making it difficult to compare studies and develop broader patterns regarding the role of transposable elements during infection. Here, we use the tools and resources available in the model, Drosophila melanogaster , to analyze multiple gene expression datasets of flies subject to bacterial, fungal, and viral infections. We analyzed differences in pathogen species, host genotype, host tissue, and sex to understand how these factors impact transposable element expression during infection. Our results highlight both shared and unique transposable element expression patterns between pathogens and suggest a larger effect of pathogen factors over host factors for influencing transposable element expression. [ABSTRACT FROM AUTHOR]

Replicated secondary contact zones can provide insights into the barriers to gene flow that are important during speciation and can reveal to which degree secondary contact may result in similar evolutionary outcomes. Here, we studied two secondary contact zones between highly differentiated Alpine butterflies of the genus Erebia using whole‐genome resequencing data. We assessed the genomic relationships between populations and species and found hybridization to be rare, with no to little current or historical introgression in either contact zone. There are large similarities between contact zones, consistent with an allopatric origin of interspecific differentiation, with no indications for ongoing reinforcing selection. Consistent with expected reduced effective population size, we further find that scaffolds related to the Z‐chromosome show increased differentiation compared to the already high levels across the entire genome, which could also hint towards a contribution of the Z chromosome to species divergence in this system. Finally, we detected the presence of the endosymbiont Wolbachia, which can cause reproductive isolation between its hosts, in all E. cassioides, while it appears to be fully or largely absent in contact zone populations of E. tyndarus. We discuss how this rare pattern may have arisen and how it may have affected the dynamics of speciation upon secondary contact. [ABSTRACT FROM AUTHOR]

Spider mites are important pests of cotton, a vital fiber crop cultivated in sun-drenched regions. This study investigated the prevalence and distribution of spider mite species infesting cotton fields in Aydin province, Türkiye. Spider mites were found in a significant portion (89%) of the surveyed fields. The two most abundant mite species, were the red form (RF) of Tetranychus urticae (57.1%) and T. turkestani (39.3%). A minor presence of T. urticae green form (GF) (3.6%) was observed. Tetranychus urticae RF was more prevalent in coastal areas, while T. turkestani was more common inland. Field surveys demonstrated that areas with higher T. urticae RF prevalence exhibited more frequent spraying, potentially due to this species’ greater capacity to develop resistance compared to T. turkestani. This study also investigated the prevalence of Wolbachia, Rickettsia, Cardinium and Spiroplasma endosymbionts, which are linked with pesticide resistance due to their ability to degrade pesticides, in these mite populations. The study confirmed the presence of Wolbachia and Rickettsia endosymbionts in spider mite populations in Türkiye. However, no Cardinium or Spiroplasma were detected in any of the populations. Infection rates of the detected endosymbionts differed between T. urticae and T. turkestani. Future studies should investigate the resistance levels of these tetranychid mite species as well as the role of their endosymbiotic bacteria in resistance in cotton-growing areas. [ABSTRACT FROM AUTHOR]

Abstract Background Mosquitoes serve as vectors for numerous pathogens, posing significant health risks to humans and animals. Understanding the complex interactions within mosquito microbiota is crucial for deciphering vector-pathogen dynamics and developing effective disease management strategies. Here, we investigated the nested patterns of Wolbachia endosymbionts and Escherichia-Shigella within the microbiota of laboratory-reared Culex pipiens f. molestus and Culex quinquefasciatus mosquitoes. We hypothesized that Wolbachia would exhibit a structured pattern reflective of its co-evolved relationship with both mosquito species, while Escherichia-Shigella would display a more dynamic pattern influenced by environmental factors. Results Our analysis revealed different microbial compositions between the two mosquito species, although some microorganisms were common to both. Network analysis revealed distinct community structures and interaction patterns for these bacteria in the microbiota of each mosquito species. Escherichia-Shigella appeared prominently within major network modules in both mosquito species, particularly in module P4 of Cx. pipiens f. molestus, interacting with 93 nodes, and in module Q3 of Cx. quinquefasciatus, interacting with 161 nodes, sharing 55 nodes across both species. On the other hand, Wolbachia appeared in disparate modules: module P3 in Cx. pipiens f. molestus and a distinct module with a single additional taxon in Cx. quinquefasciatus, showing species-specific interactions and no shared taxa. Through computer simulations, we evaluated how the removal of Wolbachia or Escherichia-Shigella affects network robustness. In Cx. pipiens f. molestus, removal of Wolbachia led to a decrease in network connectivity, while Escherichia-Shigella removal had a minimal impact. Conversely, in Cx. quinquefasciatus, removal of Escherichia-Shigella resulted in decreased network stability, whereas Wolbachia removal had minimal effect. Conclusions Contrary to our hypothesis, the findings indicate that Wolbachia displays a more dynamic pattern of associations within the microbiota of Culex pipiens f. molestus and Culex quinquefasciatus mosquitoes, than Escherichia-Shigella. The differential effects on network robustness upon Wolbachia or Escherichia-Shigella removal suggest that these bacteria play distinct roles in maintaining community stability within the microbiota of the two mosquito species.

Abstract Myrmecophilus acervorum, previously considered a parthenogenetic species widely-distributed in Europe, has been observed to have both sexes in populations inhabiting the central part of the distribution range. Specimens from those heterosexual populations have been found being infected with Wolbachia. New mitochondrial data (COI and 16S markers) revealed the well-supported differentiation of M. acervorum populations inhabiting western Polesie (Poland) and southern Europe. In turn, analyses of EF1α marker support the hypothesis on the unfinished lineage sorting at the nuclear DNA level. Interestingly, we found that parthenogenetic populations inhabiting western Polesie are infected with Wolbachia belonging to supergroup A, while endosymbionts occurring in sexual populations of M. acervorum observed in Romania belong to supergroup B. Furthermore, new and potentially diagnostic characteristics in the external structures of the eyes of M. acervorum were identified. The surface of ommatidia in specimens occurring in southern Europe was smooth. In contrast, the ommatidia surface of individuals collected in Poland was visibly sculptured. To sum up, the significant genetic variability found in the present case, and the differentiating morphological character, are almost certainly effects of cryptic species being present within M. acervorum. This is indicative of ongoing speciation within the populations of this insect, and of simultaneous unfinished lineage sorting at the nuclear DNA level.

Objetivo. Identificar e sintetizar evidências sobre efetividade e custo-efetividade da metodologia Aedes com Wolbachia para redução dos casos de arboviroses. Método. Revisão sistemática rápida, com busca realizada em fevereiro de 2022 em cinco portais de busca e bases de dados bibliográficas online. As etapas de seleção foram conduzidas por dois revisores, com resolução de conflitos por um terceiro. A extração dos dados foi feita por um revisor e conferida posteriormente por outro. Os critérios de inclusão foram estudos com descrição de custo-efetividade e efetividade da metodologia Aedes com Wolbachia, a partir de intervenções em campo, publicados nos idiomas português, inglês e espanhol, sem restrição de data de publicação. Resultados. Foram incluídos quatro estudos, sendo um ensaio randomizado em cluster, dois quase- experimental e um antes-depois, publicados entre 2019 e 2021 em quatro países. Os estudos encontrados tiveram resultados efetivos em alguns dos desfechos avaliados, como na diminuição da incidência de casos de dengue, Zika e chikungunya. Apesar de não terem sido identificados estudos experimentais abordando custo-efetividade da estratégia Aedes com Wolbachia, alguns estudos de modelagem demonstraram um potencial custo-benefício desta estratégia. Conclusão. Considerando os resultados apresentados, conclui-se que a Wolbachia apresenta potencial para ser uma estratégia economicamente efetiva e que leva à redução da incidência de casos de dengue, Zika e chikungunya. Apesar disso, não é possível, todavia, recomendar a estratégia como política pública, pois ainda são necessários mais estudos de grande porte e de alta qualidade metodológica para subsidiar a tomada de decisão política.

Moringa oleifera Lam. (moringa) is a plant native to India, used as a nutritional and medicinal supplement in many cultures around the world. Moringa has been linked to maintaining metabolic homeostasis and is often marketed as a weight loss supplement and a potential remedy for diseases such as diabetes. Here, we investigate how moringa, a ‘superfood’ with predicted protective effects against chronic diseases such as diabetes, influences the nutritional physiology and microbiome composition of the fruit fly Drosophila melanogaster. We administered moringa as a dietary supplement to Drosophila, and quantified key nutritional indices: glucose, triacylglyceride, and protein levels, and fly weight. We showed that dietary moringa supplementation significantly reduced fly glucose levels by up to ~30% and resulted in substantial restructuring of Drosophila microbiota composition, altering both gut and intracellular bacterial populations. The effect of moringa on fly glucose levels is specific because other nutritional indices, namely, triacylglyceride and protein levels and fly weight, were not significantly affected by dietary moringa supplementation. This study highlights the importance of moringa as a modulator of host glucose metabolism.

Abstract Background Tsetse flies, the biological vectors of African trypanosomes, have established symbiotic associations with different bacteria. Their vector competence is suggested to be affected by bacterial endosymbionts. The current study provided the prevalence of three tsetse symbiotic bacteria and trypanosomes in Glossina species from Burkina Faso. Results A total of 430 tsetse flies were captured using biconical traps in four different collection sites around Bobo-Dioulasso (Bama, Bana, Nasso, and Peni), and their guts were removed. Two hundred tsetse were randomly selected and their guts were screened by PCR for the presence of Sodalis glossinidius, Spiroplasma sp., Wolbachia and trypanosomes. Of the 200 tsetse, 196 (98.0%) were Glossina palpalis gambiensis and 4 (2.0%) Glossina tachinoides. The overall symbiont prevalence was 49.0%, 96.5%, and 45.0%, respectively for S. glossinidius, Spiroplasma and Wolbachia. Prevalence varied between sampling locations: S. glossinidius (54.7%, 38.5%, 31.6%, 70.8%); Spiroplasma (100%, 100%, 87.7%, 100%); and Wolbachia (43.4%, 38.5%, 38.6%, 70.8%), respectively in Bama, Bana, Nasso and Peni. Noteworthy, no G. tachnoides was infected by S. glossinidius and Wolbachia, but they were all infected by Spiroplasma sp. A total of 196 (98.0%) harbored at least one endosymbionts. Fifty-five (27.5%) carried single endosymbiont. Trypanosomes were found only in G. p. gambiensis, but not G. tachinoides. Trypanosomes were present in flies from all study locations with an overall prevalence of 29.5%. In Bama, Bana, Nasso, and Peni, the trypanosome infection rate was respectively 39.6%, 23.1%, 8.8%, and 37.5%. Remarkably, only Trypanosoma grayi was present. Of all trypanosome-infected flies, 55.9%, 98.3%, and 33.9% hosted S. glossinidius, Spiroplasma sp and Wolbachia, respectively. There was no association between Sodalis, Spiroplasma and trypanosome presence, but there was a negative association with Wolbachia presence. We reported 1.9 times likelihood of trypanosome absence when Wolbachia was present. Conclusion This is the first survey reporting the presence of Trypanosoma grayi in tsetse from Burkina Faso. Tsetse from these localities were highly positive for symbiotic bacteria, more predominantly with Spiroplasma sp. Modifications of symbiotic interactions may pave way for disease control.

Abstract The mosquito microbiome significantly influences vector competence, including in Aedes albopictus, a globally invasive vector. Describing the microbiome and Wolbachia strains of Ae. albopictus from different regions can guide area-specific control strategies. Mosquito samples from Spain and São Tomé were analyzed using 16S rRNA gene sequencing and metagenomic sequencing. Wolbachia infection patterns were observed by sex and population. Female mosquitoes were blood-fed, a factor considered in analyzing their microbiota. Results revealed a dominance of dual Wolbachia infections, strains A and B, in the microbiome of both populations of Ae. albopictus, especially among females. Both populations shared a core microbiome, although 5 and 9 other genera were only present in Spain and São Tomé populations, respectively. Genera like Pelomonas and Nevskia were identified for the first time in Aedes mosquitoes. This study is the first to describe the Ae. albopictus bacteriome in Spain and São Tomé, offering insights for the development of targeted mosquito control strategies. Understanding the specific microbiome composition can help in designing more effective interventions, such as microbiome manipulation and Wolbachia-based approaches, to reduce vector competence and transmission potential of these mosquitoes.

This study investigated the usefulness of a two-serotype dengue mathematical model to gain insights into the effects of antibody-dependent enhancement and temperature on dengue transmission dynamics in the presence of vaccination and Wolbachia-carrying mosquitoes. In particular, the effects of temperature on the mosquito death and maturation rates in secondary infections were examined. A deterministic mathematical model was formulated and analysed to address this problem. The results suggest that controlling the population of aquatic mosquitoes is appropriate for reducing the incidence of secondary infections. Furthermore, the wAu Wolbachia strain was more effective in reducing secondary infections.

Releasing Wolbachia-infected male mosquitoes to suppress wild females through cytoplasmic incompatibility (CI) has proven to be a safe and effective method for mosquito control. We study a release strategy that males are released with a constant amount c at time t k = k T , k = 0 , 1 , 2 , … . The determination of T is related to the sexually active lifespan of released males T ¯ . By introducing the early lethal effect of CI, we formulate a delay differential equation model for mosquito suppression based on Beverton-Holt-type of birth. After explicitly expressing two thresholds c ∗ and T ∗ , we prove that when c ∈ (0 , c ∗) and T ∈ (T ¯ , T ∗) , the model displays the bistable dynamics, where mosquito population either declines to zero, or approaches a positive periodic state. The rigorous mathematical proof of the exact number of periodic solutions and their stability analyses is obtained by proving that every solution is sandwiched between two “good” solutions, whose initial functions are chosen to be the solutions of the model without delay. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

The recently created Vjosa Wild River National Park sets precedence at the European scale to protect a rare river ecosystem and, at the same time, the last refuge of the almost extinct mayfly Prosopistoma pennigerum (Müller 1785, Fig. 1). We extensively searched for the species in the Vjosa and five abiotically similar but fragmented surrounding rivers. To characterize the genetic diversity of the Vjosa population and assess its potential as a source population, we analysed partial mtCOI data. Besides high abundances in the Vjosa River, we observed only a few specimens in the free-flowing section of the Osumi River, but in no other investigated river although suitable habitat appears to be widely available at site-scale. Molecular analysis of 174 specimens revealed largely homogeneous sequences across all sampling sites and an almost 100% infection rate with Wolbachia sp. that suggests parthenogenetic reproduction, possibly as a strategy to increase resilience to high natural disturbance dynamics. We argue that P. pennigerum should serve as a flagship species for the Vjosa Wild River National Park. The species is exceptionally aesthetic, and its most viable, but apparently parthenogenetic populations are restricted to the Vjosa River where the species is widespread. The restricted regional distribution indicates only weak (or no) potential to recolonize and/or re-establish in other rivers, and the absence from fragmented rivers suggests dependence on high ecological integrity. Implications for conservation: Our findings thus sanction our nomination of P. pennigerum as the flagship species for the Vjosa Wild River National Park. [ABSTRACT FROM AUTHOR]

Heritable endosymbionts widely occur in arthropod and nematode hosts. Among these endosymbionts, Wolbachia has been extensively detected in many arthropods, such as insects and crustaceans. Maternal inheritance is the most basic and dominant mode of transmission of Wolbachia, and it might regulate the reproductive system of the host in four ways: feminization, parthenogenesis, male killing, and cytoplasmic incompatibility. There is a relatively high percentage (10%) of thelytokous species in Oribatida, a suborder under the subclass Acari of arthropods, but the study of the endosymbionts in oribatid mites is almost negligible. In this paper, we detected endosymbiotic bacteria in two parthenogenetic oribatid species, Nothrus anauniensis Canestrini and Fanzago, 1877, which has never been tested for endosymbionts, and Oppiella nova, in which Wolbachia and Cardinium have been reported before. The results showed that Wolbachia was first found in N. anauniensis with an infection rate of 100% across three populations. Phylogenetic analysis showed that Wolbachia in N. anauniensis belonged to the supergroup K, marking the second supergroup of Wolbachia found in oribatid mites. Unlike previous studies, our study did not detect Wolbachia in O. nova, leading to the exclusion of Wolbachia's role in mediating thelytoky in this species. [ABSTRACT FROM AUTHOR]

Piroplasmosis, a disease of domestic and wild animals, is caused by tick-borne protozoa of the genera Babesia and Theileria, while anaplasmosis is caused by tick-borne bacteria of genera Anaplasma. Hyalomma dromedarii is the most dominant tick species infesting camels in Egypt and act as a vector of piroplasms, Anaplasma, Rickettsia and Ehrlichia spp. The available information concerning the detection of these pathogens in H. dromedarii infesting camels is limited. The present study aimed to evaluate the status of these pathogens in H. dromedarii ticks over four seasons of a year, in addition to investigate the infections of piroplasms and Anaplasmataceae besides their genetic diversity starting from June 2021 till April 2022. A total of 275 semi-engorged females of H. dromedarii were collected from different slaughtered camels, Toukh city slaughterhouse then investigated by Polymerase Chain Reaction (PCR) to detect piroplasms (Babesia spp., Theileria spp.) and Anaplasmataceae DNA targeting 18 S rRNA and 16 S rRNA genes, respectively followed by sequencing and phylogenetic analyses. Overall, piroplasms were detected in 38 ticks (13.8%), Babesia spp. was detected in 35 ticks (12.7%), while Theileria spp. was detected in one tick (0.4%). Anaplasmataceae was detected in 57 ticks (20.7%). Mixed infections of piroplasms and Anaplasmataceae were detected in 13 ticks (5%). Single infection either with piroplasms or Anaplasmataceae was detected in 25 (9%) and 44 (16%) ticks, respectively. The highest monthly rate of piroplasms was in April (spring) and Anaplasmataceae was in July (summer). Sequence analysis revealed that Babesia bigemina, Wolbachia spp. and Anaplasma marginale are the most dominant species in the examined tick samples. To the best of our knowledge, this study confirms the presence of B. bigemina, Wolbachia spp. and A. marginale in H. dromedarii in Egypt by sequencing. [ABSTRACT FROM AUTHOR]

Abstract Background Wolbachia symbiosis in Aedes aegypti is an emerging biocontrol measure against dengue. However, assessing its real-world efficacy is challenging due to the non-randomised, field-based nature of most intervention studies. This research re-evaluates the spatial–temporal impact of Wolbachia interventions on dengue incidence using a large battery of quasi-experimental methods and assesses each method’s validity. Methods A systematic search for Wolbachia intervention data was conducted via PubMed. Efficacy was reassessed using commonly-used quasi-experimental approaches coupled with extensive robustness checks, including geospatial placebo tests and a simulation study. Intervention efficacies across multiple study sites were computed using high-resolution aggregations to examine heterogeneities across sites and study periods. We further designed a stochastic simulation framework to assess the methods’ ability to reproduce true intervention efficacies (IE). Results Wolbachia interventions in Singapore, Malaysia, and Brazil were associated to significant decreases in dengue incidence, with reductions ranging from 48.17% to 69.19%. IEs varied with location and duration. Malaysia showed increasing efficacy over time, while Brazil exhibited initial success with subsequent decline, hinting at operational challenges. Singapore's strategy was highly effective despite partial saturation. Simulations identified Synthetic Control Methods (SCM) and its variant, count Synthetic Control Method (cSCM), as superior in precision, with the smallest percentage errors in efficacy estimation. These methods also demonstrated robustness in placebo tests. Conclusions Wolbachia interventions exhibit consistent protective effects against dengue. SCM and cSCM provided the most precise and robust estimates of IEs, validated across simulated and real-world settings.

Abstract Wolbachia is an obligate intracellular α-proteobacterium, which commonly infects arthropods and filarial nematodes. Different strains of Wolbachia are capable of a wide range of regulatory manipulations in their diverse hosts, including the modulation of host cellular differentiation to influence host reproduction. The genetic basis for the majority of these phenotypes is unknown. The wWil strain from the neotropical fruit fly, Drosophila willistoni, exhibits a remarkably high affinity for host germline-derived cells relative to the somatic cells. This trait could be leveraged for understanding how Wolbachia influences the host germline and for controlling host populations in the field. To further the use of this strain in biological and biomedical research, we sequenced the genome of the wWil strain isolated from host cell culture cells. Here, we present the first high quality Nanopore assembly of wWil, the Wolbachia endosymbiont of D. willistoni. Our assembly resulted in a circular genome of 1.27 Mb with a BUSCO completeness score of 99.7%. Consistent with other insect-associated Wolbachia strains, comparative genomic analysis revealed that wWil has a highly mosaic genome relative to the closely related wMel and wAu strains from Drosophila melanogaster and Drosophila simulans, respectively.