Wolbachia-induced reproductive parasitism and applications

Οι συμβιωτικές σχέσεις εντόμων-μικροοργανισμών (και ιδιαίτερα βακτηρίων) είναι ένα ευρέως διαδεδομένο φαινόμενο με ποικίλες συνέπειες στη βιολογία του ξενιστή, θετικές ή αρνητικές. Ένα από τα πιο διαδεδομένα και καλά μελετημένα βακτήρια είναι το βακτήριο Wolbachia. Το βακτήριο Wolbachia είναι ένα υποχρεωτικά ενδοκυττάριο και μητρικά κληρονομούμενο βακτήριο που συμβιώνει με ένα μεγάλο εύρος ειδών ασπόνδυλων όπως ακάρεα, αράχνες, ισόποδα, νηματώδεις της φιλαρίασης και κυρίως έντομα. Μελέτες έχουν δείξει ότι το βακτήριο Wolbachia πιθανώς συμβιώνει με το 70% όλων των ειδών των εντόμων, καθιστώντας το ως τον πιο διαδεδομένο συμβιωτικό οργανισμό του πλανήτη. Οι αλληλεπιδράσεις του βακτηρίου με τον ξενιστή κυμαίνονται μεταξύ της αμοιβαίας συμβίωσης και της επαγωγής αναπαραγωγικών ανωμαλιών όπως θηλυκοποίηση, παρθενογένεση, θανάτωση των αρσενικών ατόμων ή κυτταροπλασματική ασυμβατότητα. Παρά την ευρεία εξάπλωση και τις επιδράσεις του βακτηρίου στη βιολογία του ξενιστή, στην ειδογένεση και στην οικολογική ποικιλότητα, λίγα είναι γνωστά για τους μοριακούς μηχανισμούς της αλληλεπίδρασης του ξενιστή με το συμβιωτικό αυτό βακτήριο. Πρόσφατες μελέτες εστιάζουν στη δυνατότητα ανάπτυξης μεθόδων βιολογικού ελέγχου εντόμων γεωργικής, περιβαλλοντικής ή ιατρικής σημασίας, οι οποίες βασίζονται στις ιδιότητες του βακτηρίου Wolbachia.Insects have been reported to be associated with a broad variety of microorganisms, affecting the host biology in many different ways. Among them, Wolbachia, an obligatory intracellular and maternally-inherited symbiont, has recently attracted a lot of attention. Beside insects, Wolbachia are found in association with a wide variety of other invertebrate species, including mites, scorpions, spiders, crustaceans, filarial nematodes. Several surveys have indicated that Wolbachia may be symbiont of up to 70% of all insect species, rendering Wolbachia the most ubiquitous intracellular symbiotic organism on Earth. Wolbachia-host interactions range from many forms of reproductive parasitism to mutualistic symbioses. Different Wolbachia strains have been found to induce a number of reproductive alterations such as feminization, parthenogenesis, male-killing or cytoplasmic incompatibility. Despite their common occurrence and major effects on host biology, speciation and ecological diversity, little is known on the molecular mechanisms that mediate Wolbachia-host interactions. Recent studies focus on the potential of Wolbachia-based methods for the biological control of insect pests and disease vectors of agricultural, environmental and medical importance

Comparative microbiomics of tephritid frugivorous pests (Diptera : Tephritidae) from the field : a tale of high variability across and within species

The family Tephritidae includes some of the most notorious insect pests of agricultural and horticultural crops in tropical and sub-tropical regions. Despite the interest in the study of their gut microbiome, our present knowledge is largely based on the analysis of laboratory strains. In this study, we present a first comparative analysis of the gut microbiome profiles of field populations of ten African and Mediterranean tephritid pests. For each species, third instar larvae were sampled from different locations and host fruits and compared using 16S rRNA amplicon sequencing and a multi-factorial sampling design. We observed considerable variation in gut microbiome diversity and composition both between and within fruit fly species. A "core" microbiome, shared across all targeted species, could only be identified at most at family level (Enterobacteriaceae). At genus level only a few bacterial genera (Klebsiella,Enterobacter, andBacillus) were present in most, but not all, samples, with high variability in their relative abundance. Higher relative abundances were found for seven bacterial genera in five of the fruit fly species considered. These wereErwiniainBactrocera oleae,LactococcusinB. zonata,ProvidenciainCeratitis flexuosa,Klebsiella, andRahnellainC. podocarpiandAcetobacterandSerratiainC. rosa. With the possible exception ofC. capitataandB. dorsalis(the two most polyphagous species considered) we could not detect obvious relationships between fruit fly dietary breadth and microbiome diversity or abundance patterns. Similarly, our results did not suggest straightforward differences between the microbiome profiles of species belonging to Ceratitisand the closely relatedBactrocera/Zeugodacus. These results provide a first comparative analysis of the gut microbiomes of field populations of multiple economically relevant tephritids and provide base line information for future studies that will further investigate the possible functional role of the observed associations

Combining sterile and incompatible insect techniques for the population suppression of Drosophila suzukii

International audienceThe spotted wing Drosophila, Drosophila suzukii, has recently invaded Europe and the Americas, and it is a major threat for a wide variety of commercial soft fruits both in open field and greenhouse production systems. D. suzukii infests a wide range of ripening fruits, leading to substantial yield and revenue losses. As the application of insecticides close to the harvest period poses great concerns, the development of an efficient environment-friendly control approach to fight D. suzukii is necessary. In this study, we exploited the sterile insect technique (SIT) in combination with Wolbachia symbiosis as a population suppression approach that can constitute a potential component of an area-wide integrated pest management program. We aimed to establish a combined SIT/incompatible insect technique (IIT) protocol that would require lower irradiation doses as a complementary tool for D. suzukii management. Two D. suzukii lines trans-infected with the Wolbachia wHa and wTei strains were irradiated at doses four times less than usual (e.g., 45 Gy), and the egg hatching and adult emergence were determined. Our results indicated that wHa and wTei females as well as wHa males were sterile at this low dose. The longevity, adult emergence and flight ability of adults were evaluated, and no major effect caused by irradiation was detected. Our data indicate that a SIT/IIT protocol can be a competent approach for D. suzukii management

Trans-generational transmission of the Glossina pallidipes hytrosavirus depends on the presence of a functional symbiome

The vertically transmitted endosymbionts (Sodalis glossinidius and Wigglesworthia glossinidia) of the tsetse fly (Diptera: Glossinidae) are known to supplement dietary deficiencies and modulate the reproductive fitness and the defense system of the fly. Some tsetse fly species are also infected with the bacterium, Wolbachia and with the Glossina hytrosavirus (GpSGHV). Laboratory-bred G. pallidipes exhibit chronic asymptomatic and acute symptomatic GpSGHV infection, with the former being the most common in these colonies. However, under as yet undefined conditions, the asymptomatic state can convert to the symptomatic state, leading to detectable salivary gland hypertrophy (SGH+) syndrome. In this study, we investigated the interplay between the bacterial symbiome and GpSGHV during development of G. pallidipes by knocking down the symbionts with antibiotic. Intrahaemocoelic injection of GpSGHV led to high virus titre (109 virus copies), but was not accompanied by either the onset of detectable SGH+, or release of detectable virus particles into the blood meals during feeding events. When the F1 generations of GpSGHV-challenged mothers were dissected within 24 h post-eclosion, SGH+ was observed to increase from 4.5% in the first larviposition cycle to >95% in the fourth cycle. Despite being sterile, these F1 SGH+ progeny mated readily. Removal of the tsetse symbiome, however, suppressed transgenerational transfer of the virus via milk secretions and blocked the ability of GpSGHV to infect salivary glands of the F1 progeny. Whereas GpSGHV infects and replicates in salivary glands of developing pupa, the virus is unable to induce SGH+ within fully differentiated adult salivary glands. The F1 SGH+ adults are responsible for the GpSGHV-induced colony collapse in tsetse factories. Our data suggest that GpSGHV has co-evolved with the tsetse symbiome and that the symbionts play key roles in the virus transmission from mother to progeny

Steady states in a structured epidemic model with Wentzell boundary condition

We introduce a nonlinear structured population model with diffusion in the state space. Individuals are structured with respect to a continuous variable which represents a pathogen load. The class of uninfected individuals constitutes a special compartment that carries mass, hence the model is equipped with generalized Wentzell (or dynamic) boundary conditions. Our model is intended to describe the spread of infection of a vertically transmitted disease, for example Wolbachia in a mosquito population. Therefore the (infinite dimensional) nonlinearity arises in the recruitment term. First we establish global existence of solutions and the Principle of Linearised Stability for our model. Then, in our main result, we formulate simple conditions, which guarantee the existence of non-trivial steady states of the model. Our method utilizes an operator theoretic framework combined with a fixed point approach. Finally, in the last section we establish a sufficient condition for the local asymptotic stability of the positive steady state

Wolbachia Prophage DNA Adenine Methyltransferase Genes in Different Drosophila-Wolbachia Associations

Wolbachia is an obligatory intracellular bacterium which often manipulates the reproduction of its insect and isopod hosts. In contrast, Wolbachia is an essential symbiont in filarial nematodes. Lately, Wolbachia has been implicated in genomic imprinting of host DNA through cytosine methylation. The importance of DNA methylation in cell fate and biology calls for in depth studing of putative methylation-related genes. We present a molecular and phylogenetic analysis of a putative DNA adenine methyltransferase encoded by a prophage in the Wolbachia genome. Two slightly different copies of the gene, met1 and met2, exhibit a different distribution over various Wolbachia strains. The met2 gene is present in the majority of strains, in wAu, however, it contains a frameshift caused by a 2 bp deletion. Phylogenetic analysis of the met2 DNA sequences suggests a long association of the gene with the Wolbachia host strains. In addition, our analysis provides evidence for previously unnoticed multiple infections, the detection of which is critical for the molecular elucidation of modification and/or rescue mechanism of cytoplasmic incompatibility

The Native Wolbachia Endosymbionts of Drosophila melanogaster and Culex quinquefasciatus Increase Host Resistance to West Nile Virus Infection

The bacterial endosymbiont Wolbachia pipientis has been shown to increase host resistance to viral infection in native Drosophila hosts and in the normally Wolbachia-free heterologous host Aedes aegypti when infected by Wolbachia from Drosophila melanogaster or Aedes albopictus. Wolbachia infection has not yet been demonstrated to increase viral resistance in a native Wolbachia-mosquito host system.In this study, we investigated Wolbachia-induced resistance to West Nile virus (WNV; Flaviviridae) by measuring infection susceptibility in Wolbachia-infected and Wolbachia-free D. melanogaster and Culex quinquefasciatus, a natural mosquito vector of WNV. Wolbachia infection of D. melanogaster induces strong resistance to WNV infection. Wolbachia-infected flies had a 500-fold higher ID50 for WNV and produced 100,000-fold lower virus titers compared to flies lacking Wolbachia. The resistance phenotype was transmitted as a maternal, cytoplasmic factor and was fully reverted in flies cured of Wolbachia. Wolbachia infection had much less effect on the susceptibility of D. melanogaster to Chikungunya (Togaviridae) and La Crosse (Bunyaviridae) viruses. Wolbachia also induces resistance to WNV infection in Cx. quinquefasciatus. While Wolbachia had no effect on the overall rate of peroral infection by WNV, Wolbachia-infected mosquitoes produced lower virus titers and had 2 to 3-fold lower rates of virus transmission compared to mosquitoes lacking Wolbachia.This is the first demonstration that Wolbachia can increase resistance to arbovirus infection resulting in decreased virus transmission in a native Wolbachia-mosquito system. The results suggest that Wolbachia reduces vector competence in Cx. quinquefasciatus, and potentially in other Wolbachia-infected mosquito vectors

Mechanisms Promoting the Long-Term Persistence of a Wolbachia Infection in a Laboratory-Adapted Population of Drosophila melanogaster

Intracellular bacteria of the genus Wolbachia are widespread endosymbionts across diverse insect taxa. Despite this prevalence, our understanding of how Wolbachia persists within populations is not well understood. Cytoplasmic incompatibility (CI) appears to be an important phenotype maintaining Wolbachia in many insects, but it is believed to be too weak to maintain Wolbachia in Drosophila melanogaster, suggesting that Wolbachia must also have other effects on this species. Here we estimate the net selective effect of Wolbachia on its host in a laboratory-adapted population of D. melanogaster, to determine the mechanisms leading to its persistence in the laboratory environment. We found i) no significant effects of Wolbachia infection on female egg-to-adult survival or adult fitness, ii) no reduced juvenile survival in males, iii) substantial levels of CI, and iv) a vertical transmission rate of Wolbachia higher than 99%. The fitness of cured females was, however, severely reduced (a decline of 37%) due to CI in offspring. Taken together these findings indicate that Wolbachia is maintained in our laboratory environment due to a combination of a nearly perfect transmission rate and substantial CI. Our results show that there would be strong selection against females losing their infection and producing progeny free from Wolbachia

Towards a Wolbachia Multilocus Sequence Typing system : discrimination of Wolbachia strains present in Drosophila species

Author Posting. © The Author(s), 2006. This is the author's version of the work. It is posted here by permission of Springer for personal use, not for redistribution. The definitive version was published in Current Microbiology 53 (2006): 388-395, doi:10.1007/s00284-006-0054-1.Among the diverse maternally inherited symbionts in arthropods, Wolbachia are the most common and infect over 20% of all species. In a departure from traditional genotyping or phylogenetic methods relying on single Wolbachia genes, the present study represents an initial Multilocus Sequence Typing (MLST) analysis to discriminate closely related Wolbachia pipientis strains, and additional data on sequence diversity in Wolbachia. We report new phylogenetic characterization of four genes (aspC, atpD, sucB and pdhB), and provide an expanded analysis of markers described in previous studies (16S rDNA, ftsZ, groEL, dnaA and gltA). MLST analysis of the bacterial strains present in sixteen different Drosophila-Wolbachia associations detected four distinct clonal complexes that also corresponded to maximum-likelihood identified phylogenetic clades. Among the sixteen associations analyzed, six could not be assigned to MLST clonal complexes and were also shown to be in conflict with relationships predicted by maximum-likelihood phylogenetic inferences. The results demonstrate the discriminatory power of MLST for identifying strains and clonal lineages of Wolbachia and provide a robust foundation for studying the ecology and evolution of this widespread endosymbiont.This work was partially supported by intramural funds of the University of Ioannina to K. Bourtzis, by grants to J.J. Wernegreen from the National Institutes of Health (R01 GM62626-01) and the NASA Astrobiology Institute (NNA04CC04A), and to J.H. Werren and J.J. Wernegreen from the National Science Foundation (EF-0328363)

Wolbachia in the flesh: symbiont intensities in germ-line and somatic tissues challenge the conventional view of Wolbachia transmission routes

Symbionts can substantially affect the evolution and ecology of their hosts. The investigation of the tissue-specific distribution of symbionts (tissue tropism) can provide important insight into host-symbiont interactions. Among other things, it can help to discern the importance of specific transmission routes and potential phenotypic effects. The intracellular bacterial symbiont Wolbachia has been described as the greatest ever panzootic, due to the wide array of arthropods that it infects. Being primarily vertically transmitted, it is expected that the transmission of Wolbachia would be enhanced by focusing infection in the reproductive tissues. In social insect hosts, this tropism would logically extend to reproductive rather than sterile castes, since the latter constitute a dead-end for vertically transmission. Here, we show that Wolbachia are not focused on reproductive tissues of eusocial insects, and that non-reproductive tissues of queens and workers of the ant Acromyrmex echinatior, harbour substantial infections. In particular, the comparatively high intensities of Wolbachia in the haemolymph, fat body, and faeces, suggest potential for horizontal transmission via parasitoids and the faecal-oral route, or a role for Wolbachia modulating the immune response of this host. It may be that somatic tissues and castes are not the evolutionary dead-end for Wolbachia that is commonly thought