# Symbiont gene responsible for pest status of insect host

**Minoru Moriyama**

National Institute of Advanced Industrial Science and Technology (AIST), Tsukuba, Japan;
m-moriyama@aist.go.jp

**Takahiro Hosokawa**

National Institute of Advanced Industrial Science and Technology (AIST), Tsukuba, Japan

**Naruo Nikoh**

Department of Liberal Arts, The Open University of Japan, Chiba, Japan

**Takema Fukatsu**

National Institute of Advanced Industrial Science and Technology (AIST), Tsukuba, Japan

Understanding insect genotypes that increase or decrease pest status against agricultural plants is not only of evolutionary interest but also of applied importance. Stinkbugs of the *Megacopta cribraria-punctatissima* species complex, which are widely distributed across the East Asian region and recently invading the North America, naturally live on kudzu plants *Pueraria* spp. and often infest and damage soybean plants. Our previous study showed that *M. punctatissima* distributed in the mainland Japan successfully utilizes soybean plants whereas *M. cribraria* distributed in south-western islands of Japan does not perform well on soybean plants. Symbiont exchange experiments revealed that their gut symbiotic bacteria *Ishikawaella*, rather than the insects themselves, determine the different pest status of the insect hosts on soybean. Here we report comparative analyses of *Ishikawaella* genomes associated with these two species to determine symbiont genes relevant to the pest status. We found that these symbiont genomes are quite similar: in total, 173 site differences were identified, which consisted of base substitutions and small indels. Notably, open reading frames for two bacterial genes encoding Ary (arylesterase) and Era were disrupted only in the *Ishikawaella* genome associated with *M. cribraria*. When these two symbiont genes were genotyped in geographical populations of *M. cribraria-punctatissima* complex across the Japanese Archipelago, three types were identified: both Ary and Era are intact; Ary is intact and Era is disrupted; and both Ary and Era are disrupted. When insects from six Japanese populations, whose symbionts exhibit these different genotypes, were reared on soybean plants, their performance was correlated with intact / disrupted Ary genotype. Our finding provides insights into the mechanism and the evolutionary origin of insect pests, potentially leading to novel applications.
*Wolbachia* diversity between host phenotype and phylogenetic incongruence between *Wolbachia* and WO phage in *Vollenhovia emeryi*

**Soyeon Park**

Division of Ecoscience, Ewha Womans University, Seoul, Korea; sy.soyeon@gmail.com

**Pureum Noh**

Division of Ecoscience, Ewha Womans University, Seoul, Korea

**Yong-Jin Won**

Division of Ecoscience, Ewha Womans University, Seoul, Korea

**Jae Chun Choe**

Division of Ecoscience, Ewha Womans University, Seoul, Korea

**Gilsang Jeong**

Division of Ecoscience, Ewha Womans University, Seoul, Korea

The *Vollenhovia emeryi* ant, widely distributed in Far East, shows queen polymorphism associated with *Wolbachia* infection status. The phylogenetic analysis of the ant's mtDNA reveals derivation of the *Wolbachia*-free short-winged from the *Wolbachia*-infected long-winged. However, intriguingly, some Japanese short-winged colonies harbor *Wolbachia*. *Wolbachia* specific bacteriophage (WO) is also detected in more than half of the infected colonies with no clear distribution pattern across the host insect lineage. We hypothesized that 1) the infected Japanese short-winged is in the intermediate stage to complete loss of *Wolbachia* and 2) the phage invaded the host after the host insect diverged. To test the hypotheses, we studied the strain diversity using the multi-locus sequence typing (MLST) of five ant colonies; three long-winged colonies from Korea and one long-winged colony and one short-winged colony from Japan. Both Korean and Japanese *V. emeryi* colonies show unexpectedly high levels of *Wolbachia* strain diversity. However, the diversity is not significantly different between the long-winged and the short-winged against our first hypothesis. Phylogenies of *Wolbachia* show Korean strains and Japanese strains are largely monophyletic indicating prior infection before the host divergence. The strain diversity of the phage is also surprisingly high. Phylogenies of *orf2* and *orf7* genes are incongruent to that of *Wolbachia* and geographically distinct. This indicates that the phage is spatially static and the current infection pattern may be the consequence of local repeated gain and loss of the phage.
# What does the influence of *Wolbachia* on the testis stem cell niche reveal on the impact of the bacteria on the *Drosophila melanogaster* male biology?

**Stéphanie M. Pontier**

Department of Development and Stem cells, Institut Pasteur, Paris, France

**François Schweisguth**

Department of Development and Stem cells, Institut Pasteur, Paris, France

Stem cells and their niches are in charge of tissue homeostasis and as such constitute sensitive sentinels of the physiology of an organism. In order to study with precision how physiology affects a stem cell niche, we have developed a quantitative mathematical strategy based on the characterization of the niche cellular composition at the single niche level. By applying this strategy to the testis stem cell niche, we have discovered that infection of *D. melanogaster* with *wMelCS* affected significantly the testis stem cell niche homeostasis in just eclosed adult males. *Wolbachia* having a particular tropism for the gonad stem cell niches in several *Drosophila* species, its impact on these niches might be local. We provide evidences suggesting that local impact is unlikely to fully explain our observations in young *D. melanogaster* males. Indeed, simple manipulations of the social environment of *Wolbachia*-infected males during their metamorphosis are sufficient to restore a testis niche homeostasis comparable to the one observed in control males. In contrast, restoration of the niche homeostasis is precluded in olfactive dead mutant males infected with *wMelCS*. Finally, we show that the influence of social environment on *Wolbachia*-infected males during their metamorphosis is not restricted to the testis tissue and has systemic consequences, both at the eclosion time and the fertility levels.
# The history and ongoing invasion of *Wolbachia* in European *Rhagoletis cerasi* populations

**Hannes Schuler**

Faculty of Science and Technology, Free University of Bozen-Bolzano, Italy; Hannes.Schuler@unibz.it

**Kirsten Köppler**

Center for Agricultural Technology Augustenberg, Karlsruhe, Germany

**Sabine Daxböck-Horvath**

Department of Crop Sciences, University of Natural Resources and Life Sciences, Vienna, Austria

**Bilal Rasool**

Government College University, Faisalabad, Pakistan

**Susanne Krumböck**

Department of Forest and Soil Sciences, University of Natural Resources and Life Sciences, Vienna, Austria

**Dietmar Schwarz**

Department of Biology, Western Washington University, Bellingham, USA

**Thomas Hoffmeister**

Institute of Ecology, University of Bremen, Germany

**Birgit C. Schlick-Steiner**

Institute of Ecology, University of Innsbruck, Austria

**Florian M. Steiner**

Institute of Ecology, University of Innsbruck, Austria

**Christian Stauffer**

Department of Forest and Soil Sciences, University of Natural Resources and Life Sciences, Vienna, Austria

**Wolfgang Arthofer**

Institute of Ecology, University of Innsbruck, Austria

**Markus Riegler**

Hawkesbury Institute for the Environment, University of Western Sydney, Penrith, Australia

*Wolbachia* is a widespread reproductive parasite of insects that can impact host ecology and evolution. The European cherry fruit fly, *Rhagoletis cerasi*, is infected by up to five distinct *Wolbachia* strains. The most prevalent strains are *wCer1*, fixed in all populations, and *wCer2*, currently invading European populations by inducing cytoplasmatic incompatibility.

Here we analyze the spatial distribution of these two *Wolbachia* strains in European *R. cerasi* populations and compare it with the mitochondrial and nuclear genotypes of its host. We found heavily reduced host mitochondrial diversity, indicative for two sequential selective sweeps caused by the current *wCer2* and the former *wCer1* invasion. While both infections are associated with specific mitochondrial haplotypes, the analysis of seven microsatellite loci showed no *Wolbachia* linked nuclear structure. Detailed population data over a time period of ten years suggests that *wCer2* is rarely lost from infected populations; occasional intraspecific horizontal transmission events were detected in the transition zone in central Germany, however, without disrupting the linkage of *Wolbachia* and mitochondrial haplotypes outside this region. Based on our combined dataset we discuss the evolutionary history of *Wolbachia* in *R. cerasi*.
**Wolbachia as incompatibility factor between Rhagoletis populations from North America and Mexico**

**Hannes Schuler**

Faculty of Science and Technology, Free University of Bozen-Bolzano, Italy; Hannes.Schuler@unibz.it

**Eduardo Tadeo**

Instituto de Ecología A.C., Xalapa, Mexico

**Juan Rull**

Instituto de Ecología A.C., Xalapa, Mexico

**Martin Aluja**

Instituto de Ecología A.C., Xalapa, Mexico

**Scott P. Egan**

Department of Ecology and Evolutionary Biology, Rice University, Houston, USA

**Christian Stauffer**

Department of Forest & Soil Sciences, University of Natural Resources and Life Sciences, Vienna, Austria

**Jeffrey L. Feder**

Department of Biological Sciences, University of Notre Dame, USA

*Wolbachia* is probably the most widespread endosymbiont, infecting a broad range of arthropod species. This bacterium can have significant influences in the biology of its host by influencing its reproduction. The most common mechanism is the induction of a cytoplasmic incompatibility that can lead to a postzygotic reproductive isolation among different *Wolbachia* infected populations.

*Rhagoletis* is a serious pest in orchards infesting various different fruits mainly in North America and Mexico. The most famous member of this genus is the apple maggot *Rhagoletis pomonella*. Naturally infesting hawthorns, a population shifted to domesticated apples as a new host. This resulted in the formation of an ecologically and genetically distinct host race, which acts as a textbook example of sympatric speciation. Additionally, recent studies have shown that *R. pomonella* formed at least two genetically isolated populations in Mexico.

In contrast, speciation in the *R. cingulata* species group appears to have formed by a variety of different modes: cherry-infesting *R. cingulata* and *R. indifferens* are diverging by classic allopatric (geographic) speciation, while *R. osmanthi* and *R. chionanthi* formed sympatrically, by adapting to different hosts. Similar to *R. pomonella*, *R. cingulata* also has isolated populations in Mexico.

Mating studies crossing North American *R. pomonella* and *R. cingulata* populations with flies from Northern and Central Mexico have revealed a degree of postzygotic reproductive isolation. In this presentation, we examine the cause of the incompatibility by studying their *Wolbachia* infections and discuss the possible influence of this bacteria on the evolution of reproductive isolation for *Rhagoletis* flies.
# Pharmacokinetic / pharmacodynamic modelling of *Wolbachia* growth dynamics under antibiotic drug pressure in a lymphatic filariasis murine infection model

**Raman Sharma**

Department of Parasitology, Liverpool School of Tropical Medicine, UK; rsharma8@liv.ac.uk

**Ghaith Aljayoussi**

Department of Parasitology, Liverpool School of Tropical Medicine, UK

**Steven A. Ward**

Department of Parasitology, Liverpool School of Tropical Medicine, UK

**Mark J. Taylor**

Department of Parasitology, Liverpool School of Tropical Medicine, UK

**Ana de Castro Guimaraes**

Department of Parasitology, Liverpool School of Tropical Medicine, UK

**Jill Davies**

Department of Parasitology, Liverpool School of Tropical Medicine, UK

**Louise Ford**

Department of Parasitology, Liverpool School of Tropical Medicine, UK

**Alison Shone**

Department of Parasitology, Liverpool School of Tropical Medicine, UK

**Joseph D. Turner**

Department of Parasitology, Liverpool School of Tropical Medicine, UK

**David Waterhouse**

Department of Parasitology, Liverpool School of Tropical Medicine, UK

*Wolbachia* is an essential endosymbiont of the filarial nematodes *Wuchereria bancrofti*, *Brugia malayi* and *Onchocerca volvulus* which are the causative agents of lymphatic filariasis and onchocerciasis. An estimated 120 million are infected by lymphatic filariasis and 17.7 million infected by onchocerciasis throughout the tropics leading to a profound public health and socio-economic burden in severely affected communities.

Targeting of *Wolbachia* in these filarial nematodes using antibiotic chemotherapy has been shown to deliver a safe macrofilaricidal activity and reduce filarial burden, morbidity and transmission. The current gold standard anti-*Wolbachia* drug regimen is a 100 - 200 mg/day doxycycline dose given for four to six weeks. The A•WOL consortium funded by a grant awarded to Liverpool School of Tropical Medicine by the Bill & Melinda Gates Foundation aims to reduce the current treatment time to seven days or less as aligned with the Target Product Profiles for an anti-*Wolbachia* macrofilaricide.

To achieve a rapid seven-day or less kill rate of *Wolbachia*, a number of antibiotic combinations will be optimised. These include the tetracyclines (doxycycline and minocycline), rifamycins (rifampicin and rifapentine), and a fluoroquinolone (moxifloxacin) as well as combinations with anti-helminthic drugs. Our rational approach involves identifying optimal treatments in *in-vivo* models and translation from the lab into clinical field trials.

We have initially studied the dynamics of *Wolbachia* under drug pressure with doxycycline, minocycline, moxifloxacin, and rifampicin in the AWOL validated mouse *Brugia malayi* infection model. Using experimental pharmacokinetic (PK) and pharmacodynamic (PD) data we have constructed a series of PK-PD models and simulations to further dissect and quantify the dynamics of *Wolbachia* growth.

This data analysis displays the power of PK-PD modelling in quantifying the dynamics of *Wolbachia* growth under antibiotic drug pressure.
# Investigation of a candidate *Wolbachia* type IV effector from strain *wAna* provides insight into a mechanism for host manipulation

**Kathy B. Sheehan**

Department of Biology, Indiana University, Bloomington, USA; sheehaka@gmail.com

**Irene L. G. Newton**

Department of Biology, Indiana University, Bloomington, USA

*Wolbachia* comprises a group of phylogenetically diverse, obligately intracellular bacteria famous for manipulating a broad range of invertebrate hosts. The molecular tools that are used by these bacteria in order to control host cell biology are currently unknown. From genomic sequencing of various *Wolbachia* strains, we know that the bacteria encode a complete, type IV secretion system that likely secretes 'effectors' into host cells during infection. We utilized a combination of high-throughput screening in yeast and bioinformatics to identify and characterize a potential effector from *Wolbachia* that infects *Drosophila ananassae*. We focused on this protein because our analysis revealed that it contains a domain homologous to those found in plant apoptosis machinery. Transcripts of the protein were found in infected, but not uninfected, *D. ananassae* ovaries. Using a custom antibody targeting the putative *Wolbachia* effector, we were able to show that the full-length protein specifically expressed across developmental stages of infected flies. Additionally, we investigated the presence of the protein in cytosolic fractions of infected host cells, a property necessary for secreted effectors. We conducted a yeast-two-hybrid screen to identify interaction partners revealing interactions with important members of the host's oxidative stress response pathways. Because *Wolbachia* are known to manipulate ROS levels in their hosts, we predict that these secreted proteins may be involved in altering host cell redox state.
*In vitro* and *in vivo* assays for the assessment of cytoplasmic competence of *wDi* from the Asian citrus psyllid, *Diaphorina citri*, with other *Wolbachia* strains

Paul D. Shirk

Agricultural Research Service, United States Department of Agriculture, Gainesville, USA; paul.shirk@ars.usda.gov

Richard B. Furlong

Agricultural Research Service, United States Department of Agriculture, Gainesville, USA

Eric D. LoVullo

Agricultural Research Service, United States Department of Agriculture, Gainesville, USA

To assess how different strains of *Wolbachia* compete when occupying the same cells, superinfections of various *Wolbachia* strains were established in the same cultured cells and insects. The most prevalent *Wolbachia* strain of the Asian citrus psyllid, *Diaphorina citri*, *wDi* (Supergroup B) was transinfected into the *Bombyx mori* Bm5 cell line starting with newly laid eggs. *Wolbachia* from the Caribbean fruit fly, *Anastrepha suspensa*, *wAsu* (Supergroup A), the Mediterranean flour moth, *Ephestia kuehniella*, *wEku* (Supergroup A), *wMel* (Supergroup A) and *wAlbB* (Supergroup B) were also established in Bm5 cells. To test interactions between *wDi* and other *Wolbachia* strains, dual-infections between Bm5/*wDi* and the other *Wolbachia* strains were made. The dynamics of the dual-infections were followed using qRT-PCR to measure the copy number of each strain relative to the Bm5 host genome. Dual-infections between *wDi* and the other strains resulted in extensive Bm5 cell death within seven days.

Purified *Wolbachia* strains from the cultured cell lines were used to establish single infections in *Drosophila simulans*. Establishment of the *Wolbachia* infection was confirmed by direct PCR using strain specific primers. The single infected *D. simulans* *wDi* strain was cross-mated with each of the other single infected *D. simulans* strains to establish superinfections to assess cytoplasmic incompatibility (CI) effects. The ratio of the co-infecting *Wolbachia* was monitored by qRT-PCR. Studies established CI effects in cross-mated flies with each singular *Wolbachia* infected strain. We discuss the potential of utilizing other *Wolbachia* strains to provide CI biological drive in the Asian citrus psyllid.
**Wolbachia mediated protection against pathogenic bacteria in
two isopod models**

**Christine Braquart-Varnier**

Laboratoire Ecologie et Biologie des Interactions, Université de Poitiers, France

**Mine Altinli**

Laboratoire Ecologie et Biologie des Interactions, Université de Poitiers, France

**Romain Pigeault**

Maladies Infectieuses et Vecteurs: Écologie, Génétique, Évolution et Contrôle (MIVEGEC), Montpellier, France

**Frédéric Chevalier**

Texas Biomedical Research Institute, San Antonio, USA

**Gaëtan Mappa**

Laboratoire Ecologie et Biologie des Interactions, Université de Poitiers, France

**Pierre Grève**

Laboratoire Ecologie et Biologie des Interactions, Université de Poitiers, France

**Didier Bouchon**

Laboratoire Ecologie et Biologie des Interactions, Université de Poitiers, France

**Mathieu Sicard**

Institut des Sciences de l'Évolution, Université Montpellier 2, France; Mathieu.sicard@univ-montp2.fr

Symbionts largely affect their hosts' life history traits and fitness. Previous studies showed that vertically transmitted symbionts can protect the host against pathogens. Among this symbionts, the *Wolbachia* have recently been shown to confer protection to their hosts against a wide range of pathogens, especially viruses. The present study aims to test whether resident *Wolbachia* could confer protection to terrestrial isopods against invasive intracellular bacteria (*Salmonella typhimurium*, *Listeria ivanovii* but also the pathogenic *Wolbachia* strain *wWulC*). To do so, we assessed the survival during bacterial infections of *Armadillidium vulgare* and *Porcellio d. dilatatus* isopods when they are symbiotically associated or not with resident *Wolbachia*. We showed that when feminizing and CI *Wolbachia* have an effect on the survival of their hosts during the bacterial infection, this effect is always beneficial to the host. However, the intensity of the "anti-pathogenic" effect is clearly dependent on the *Wolbachia* strain and the genetic background of the host.
# Insights into the evolution of *Cardinium*: the development of a Multi Locus Sequence Typing system

**Corinne M. Stouthamer**

Department of Entomology, University of Arizona, Tucson, USA; cmstouthamer@gmail.com

**Marco Gebiola**

Department of Entomology, University of Arizona, Tucson, USA

**Suzanne E. Kelly**

Department of Entomology, University of Arizona, Tucson, USA

**Martha S. Hunter**

Department of Entomology, University of Arizona, Tucson, USA

*Cardinium* is an intracellular reproductive manipulator in the phylum Bacteroidetes that is found in many different species of arthropods. This independently-evolved symbiont lineage exhibits three of the four reproductive phenotypes attributed to *Wolbachia*, including cytoplasmic incompatibility, long thought to be unique to *Wolbachia*-infected hosts. Genetic and phylogenetic analyses of *Cardinium* in comparison with *Wolbachia* offer an opportunity to determine functional equivalents and differences between the two lineages. Placing individual strains of *Cardinium* within a larger evolutionary context is currently challenging because only two genes (16S rDNA and Gyrase B) have been used to generate phylogenetic trees, and consequently, the relationship of these different strains has only been elucidated in its roughest form. The development of a Multi Locus Sequence Typing (MLST) system would provide the research community with new primers for detecting *Cardinium*, unambiguous methods of delineating strains, and more informative phylogenetic trees. I am in the process of designing universal *Cardinium* PCR primers for four different genes: Translation Elongation Factor G (EF-G), Heat Shock Protein (groEL), Iron Sulfur Cluster Assembly Protein (sufB), and Gyrase B (gyrB). These will be used to identify, describe, and place strains of *Cardinium* into a phylogenetic tree. Based on my preliminary tree, *Cardinium* appears to display more host affinity than *Wolbachia*. However, similar to *Wolbachia*, the reproductive manipulations are dispersed across the tree, and thus have likely evolved independently multiple times. To date, this is the most robust tree available for investigating *Cardinium* evolution.
# The role of the host in *Wolbachia*-virus interactions

**Jakob F. Strauss**

Institute for Evolution and Biodiversity, Westfalian Wilhelms-University, Munster, Germany; jfstrauss@gmail.com

**Takeshi Miki**

Institute of Oceanography, National Taiwan University, Taipei, Taiwan

**Arndt Telschow**

Institute for Evolution and Biodiversity, Westfalian Wilhelms-University, Munster, Germany

There is growing empirical evidence that *Wolbachia* (*WB*) directly interact with viruses inside the arthropod host, sometimes resulting in low or no virus replication. Previous theoretical studies showed that this direct effect of *WB* can result in reduced virus prevalence (within the population), suggesting that *WB* could be used in the biological control of vector-borne diseases (e.g., dengue fever). However, most of the models focus on *WB*-virus interactions, whereas the potential roles of the host are overlooked. In order to fill this gap, we investigated two questions through mathematical modeling. (1) How is host resistance against the virus affected by *WB*? We investigated the tripartite interactions between *WB*, virus, and host by a model that incorporates (i) horizontal virus- and vertical *WB*-transmission, (ii) cytoplasmic incompatibility (CI), (iii) *WB* resistance against virus, and (iv) host resistance against virus. Our main finding is that the spread of *WB* causes the loss of costly host resistance alleles. We discuss implications of this loss of genetic diversity with respect to vector-borne diseases. (2) How does the host's life history affect *WB*-virus codynamics? We consider a scenario, in which *WB* do not affect virus replication inside coinfected hosts. However, *WB* might indirectly affect the virus because reproductive phenotypes (CI or male killing) increase larval mortality of hosts and thus alter virus dynamics. Our analysis revealed that this indirect effect depends strongly on the host's life history, and can result in two opposing outcomes: (i) reduced virus prevalence and virus invasion ability, and (ii) increased virus prevalence and virus invasion ability. The former occurs for host species with larval competition and undercompensation, the latter for hosts with either adult competition or larval competition and overcompensation. These findings suggest that the effect of *WB* on a specific virus is sensitive to the host's life history.
# A male-killing *Wolbachia* affects the dosage compensation of its lepidopteran host?

**Takafumi N. Sugimoto**

Frontier Research Core for the Life Sciences, University of Toyama, Japan; sugimoto@ctg.u-toyama.ac.jp

**Takumi Kayukawa**

Division of Insect Sciences, The National Institute of Agrobiological Sciences, Ibaraki, Japan

**Yukio Ishikawa**

Department of Agricultural and Environmental Biology, The University of Tokyo, Japan

**Tetsuro Shinoda**

Division of Insect Sciences, The National Institute of Agrobiological Sciences, Ibaraki, Japan

**Tsutomu Tsuchida**

Frontier Research Core for the Life Sciences, University of Toyama, Japan

*Wolbachia*, a group of endosymbiotic bacteria harbored by a wide range of insects, is known for various manipulations of host reproduction to expedite their own propagation. *Wolbachia* (*wSca*) infecting the Adzuki bean borer moth *Ostrinia scapulalis* (Lepidoptera: Crambidae) causes male killing, in which males (genotype ZZ) selectively die during embryonic and early larval development, whereas females (genotype ZW), in turn, selectively die when cured of infection.

In a previous study, we analyzed phenotypic sex (sex determination gene expression pattern, male-type or female type) and genetic sex (sex chromosome type, ZZ or ZW) of the embryos and larvae of normal, *wSca*-infected, and infected-and-cured *O. scapulalis*. As the results, it was observed that the female-type sex-determination gene was expressed in the infected genetic male (ZZ) progenies destined to die, whereas the male-type sex-determination gene was expressed in the cured genetic female (ZW) progenies destined to die. This result suggested the discordance of the genetic and phenotypic sexes underlie the sex-specific death, but the mechanism of death is unresolved. One plausible explanation for sex-dependent lethality may be discordance in dosage compensation, i.e. sex-dependent adjustment of the expression levels of genes on the sex chromosomes.

So, we focused on the dosage compensation and analyzed the Z chromosome-linked gene expression by quantitative PCR. For this conference, we will describe the result of comparison experiment of Z chromosome-linked gene expression in the embryos and larvae of normal, *wSca*-infected, and infected-and-cured *O. scapulalis*.
# Reproductive parasitism and gene flow modification

**Arndt Telschow**

Institute for Evolution and Biodiversity, Westfalian Wilhelms-University, Munster, Germany; a.telschow@wwu.de

**Yutaka Kobayashi**

Kochi University of Technology, Japan

**John H. Werren**

University of Rochester, USA

**Peter Hammerstein**

Institute for Theoretical Biology, Humboldt-Universität zu Berlin, Germany

The term reproductive parasitism is commonly used to describe infective agents that manipulate the reproductive system of their host to their own advantage. Among the reproductive manipulations are feminization, male killing, and cytoplasmic incompatibility. Well-studied reproductive parasites belong to the bacterial groups *Wolbachia*, *Rickettsia*, *Cardinia*, *Arsenophonus*, and *Spiroplasma*. The wide distribution of these bacterial groups among arthropods makes the study of reproductive parasitism an important topic in evolution. The present (theoretical) study is motivated by the (experimental) findings that different host populations often differ with respect to their infection state, i.e. some populations are infected while others are not or with a different strain. Based on these findings we constructed several population genetic models in order to investigate how such infection polymorphism affect the gene flow between populations. To measure gene flow we make use of the concept of effective migration rate. Our analysis revealed that, generally, infection with reproductive parasites modify host gene flow. Interestingly, populations with highest degree of infection are thereby converted into population genetic sinks. This is true for all forms of reproductive parasitism. These results give general insights to the evolution of selfish genetic elements. We discuss the implications with respect to host evolution and host-parasite coevolution, especially speciation and local adaptation.
# Host plant specialization matters in the epidemiology of *Wolbachia* across phytophagous wasps (Hymenoptera: Torymidae)

**Thomas Boivin**

INRA, Avignon, France

**Hélène Henri**

Biometry and Evolutionary Biology Lab, University of Lyon 1, France

**Fabrice Vavre**

Biometry and Evolutionary Biology Lab, University of Lyon 1, France; fabrice.vavre@univ-lyon1.fr

**Cindy Gidoin**

INRA, Avignon, France

**Philippe Veber**

Biometry and Evolutionary Biology Lab, University of Lyon 1, France

**Jean-Noel Candau**

Great Lakes Forestry Centre, Ontario, Canada

**Alain Roques**

INRA, Avignon, France

**Marie-Anne Auger-Rozenberg**

INRA, Avignon, France

Among eukaryotes, sexual reproduction is by far the most predominant mode of reproduction. However, some systems maintaining sexuality appear particularly labile and raise intriguing questions on the evolutionary routes to asexuality. Thelytokous parthenogenesis is a form of spontaneous loss of sexuality leading to strong distortion of sex ratio towards females and resulting from mutation, hybridization or infection by bacterial endosymbionts. We investigated whether ecological specialization is a likely mechanism of spread of thelytoky within insect communities. Focusing on the highly-specialized genus *Megastigmus* (Hymenoptera: Torymidae), we first performed a large literature survey to examine the distribution of thelytoky in these wasps across their respective obligate host plant families. Second, we tested for thelytoky caused by endosymbionts by screening in 15 arrhenotokous and 10 thelytokous species for *Wolbachia*, *Cardinium*, *Arsenophonus* and *Rickettsia* endosymbionts and by performing antibiotic treatments. Finally, we performed phylogenetic reconstructions using multilocus sequence typing (MLST) to examine the evolution of endosymbiont-mediated thelytoky in *Megastigmus* and its possible connections to host plant specialization. We demonstrate that thelytoky evolved from ancestral arrhenotoky through the horizontal transmission and the fixation of the parthenogenesis-inducing *Wolbachia*. We find that ecological specialization in *Wolbachia's* hosts was probably a critical driving force for *Wolbachia* infection and spread of thelytoky, but also a constraint. Our work further reinforces the hypothesis that community structure of insects is a major driver of the epidemiology of endosymbionts and that competitive interactions among closely related species may facilitate their horizontal transmission.
**Wolbachia infection in Polish *Liophloeodes* (Coleoptera:Curculionidae) populations: pathways of transfer and influence on host reproduction**

**Beniamin Waclawik**

Institute of Zoology, Jagiellonian University, Krakow, Poland; beniamin.waclawik@uj.edu.pl

**Dorota Lachowska-Cierlik**

Institute of Zoology, Jagiellonian University, Krakow, Poland

**Iwona Warzyszynska**

Institute of Zoology, Jagiellonian University, Krakow, Poland

*Wolbachia* routes of transmission are one of most widely discussed problem applying to this bacterium. Although significant role of vertical transfer in spreading of *Wolbachia* is confirmed, frequent incongruence between phylogenies of the bacterium and host indicates that also horizontal transmission is important. Good way to find out routes of *Wolbachia* movement among hosts from particular systematic group is to compare phylogeny of endosymbiont with host phylogeny (similarities pinpoints vertical transfer) and host ecology and geography (similarities suggest horizontal transfer).

About 40% of European weevil species are estimated to be infected with *Wolbachia*. Among them are two species from *Liophloeodes* subgenus *L. lentus* and *L. gibbus*. Both of them are Carpathian subendemic species with similar ecological preferences. In Poland there are many populations of these weevils and there are potential contact zones between divergent populations, that could enable vertical transfer.

The main goal of this research was to discover possible pathways of *Wolbachia* transfer among Polish populations of these two species. Weevils from 15 populations were tested. Their phylogeny (based on EF1-alpha gene) and geographical distribution were compared with phylogeny of their *Wolbachia* endosymbionts (based on ftsZ and wsp genes).

*Wolbachia* infections and vertical transfer cause many changes in host reproduction. These abnormalities often lead to discordances between mitochondrial and nuclear DNA phylogeny. In second part of the research, nDNA (EF1-alpha and ITS2) and mtDNA (COI and CytB) phylogenies of 20 Polish *Liophloeodes* populations were compared. There was made attempt to discover discordances that would indicate significance of vertical transfer.

Serious incongruence between mtDNA and nDNA phylogeny was found out, however *Wolbachia* phylogeny indicates great role of horizontal transfer among studied populations.
# Combining Sterile Insect Technique with *Wolbachia*-based approaches: assessing effects on fitness of triple and double *Wolbachia*-infected strains of *Aedes albopictus*

**Dongjing Zhang**

Insect Pest Control Laboratory, Joint FAO/IAEA Division of Nuclear Techniques in Food and Agriculture, Vienna, Austria; zhangdongjing06@163.com

**Zhiyong Xi**

Department of Microbiology and Molecular Genetics, Michigan State University, East Lansing, USA.

**Konstantinos Bourtzis**

Insect Pest Control Laboratory, Joint FAO/IAEA Division of Nuclear Techniques in Food and Agriculture, Vienna, Austria

**Jeremile Gilles**

Insect Pest Control Laboratory, Joint FAO/IAEA Division of Nuclear Techniques in Food and Agriculture, Vienna, Austria

Together with other control methods, the Sterile Insect Technique (SIT) and the Incompatible Insect Technique (IIT) are currently being considered, separately or in combination, as potential approaches for the suppression of populations of the major vector mosquito species *Aedes albopictus* in Guangzhou, China. Natural populations of *Ae. albopictus* are double-infected with the *Wolbachia* strains *wAlbA* and *wAlbB*. By embryonic microinjections, a new triple *Wolbachia* infected line (*wAlbA*, *wAlbB*, and *wPip*), HC line, has been developed which expresses strong Cytoplasmic Incompatibility (CI) in matings between the transinfected males and wild type females. In this study, we present the comparative analysis of the life history traits of three *Ae. albopictus* lines (triple-infected, double-infected and uninfected), all of them with the same ("Guangzhou") genomic background. Under the same rearing conditions, the egg-hatching rate, survivorship of pupae and adults, sex ratio, duration of larval stage (L1 to pupae stage), duration of adult stage (L1 to adult emergence), female fecundity and male longevity were investigated. Our results suggest a minimal effect of *Wolbachia* on the fitness of these populations. Based on this evidence, the triple-infected line is currently considered for mass rearing and the application of a combined SIT-IIT strategy to control *Ae. albopictus* populations in the Guangzhou area in China.
# Knockdown of ATPsyn-b caused larval growth defect and male infertility in *Drosophila*

**Ya-Na Chen**

College of Life Sciences, Central China Normal University, Wuhan, China

**Ya Zheng**

College of Life Sciences, Central China Normal University, Wuhan, China; zhengyahb@163.com

**Jing-Jing Li**

College of Life Sciences, Central China Normal University, Wuhan, China

**Jia-Lin Wang**

College of Life Sciences, Central China Normal University, Wuhan, China

**Yu-Feng Wang**

College of Life Sciences, Central China Normal University, Wuhan, China

ATPsyn-b gene is encoded *Drosophila* mitochondrial membrane F0F1-type ATPase F0 b subunit, involved in the process of oxidative phosphorylation and ATP synthesis, thus providing energy for *Drosophila* life activities. In this study, firstly Gal4-UAS system was adopt for acquiring flies lines with ATPsyn-b knock-down both in testis and extensive body respectively. Then, males embryo hatching were detected and showed they are infertile when ATPsyn-b knock-down occur in testis. Further study showed this male sterility was caused by abnormal spermatogenesis, such as sperm nuclei in cyst of mutant males were completely decentralized, thus failed to form normal sperm bundles. Moreover, individualized mature sperm were absent in seminal vesicles near the end of testis. Additionally, ATPsyn-b widely knock-down flies showed developmental stagnation just in the second instar larvae while the control larvae had developed into the third instar. Remarkably, ATPsyn-b knock-down individual were still remain as the size of the second instar larvae until control adult had been all emerged, and died finally. Combining the result of qRT-PCR, that proved the phenotype of arrested development was indeed caused by ATPsyn-b knock-down, which also show cell volume failed to increase proved by Brdu incorporation experiments. In short, our results proved that ATPsyn-b gene played an important role on both *Drosophila* spermatogenesis and larval development, which may help offer a new direction and clue for studying insect male infertility. Further deeper study on the mechanism of infertility were also needed.
Author index

-A-

<table><tr><td>Abd-Alla A. M. M.</td><td>48</td></tr><tr><td>Aksoy S.</td><td>56</td></tr><tr><td>Albertson R.</td><td>35</td></tr><tr><td>Aljayoussi G.</td><td>106</td></tr><tr><td>Altinli M.</td><td>109</td></tr><tr><td>Aluja M.</td><td>105</td></tr><tr><td>Anbutsu H.</td><td>71, 74</td></tr><tr><td>Armstrong S. D.</td><td>50</td></tr><tr><td>Arroyo-Yebras F.</td><td>13, 76</td></tr><tr><td>Arthofer W.</td><td>94, 104</td></tr><tr><td>Asiimwe P.</td><td>37</td></tr><tr><td>Asimakis H.</td><td>27</td></tr><tr><td>Atyame C.</td><td>61</td></tr><tr><td>Auger-Rozenberg M. A.</td><td>114</td></tr><tr><td>Augustinos A.</td><td>27</td></tr></table>

-B-

<table><tr><td>Badawi M.</td><td>66, 75</td></tr><tr><td>Bah G. S.</td><td>50</td></tr><tr><td>Bailly-Bechet M.</td><td>10</td></tr><tr><td>Bandi C.</td><td>98</td></tr><tr><td>Bauer S.</td><td>26</td></tr><tr><td>Bazzocchi C.</td><td>98</td></tr><tr><td>Bella J. L.</td><td>13, 68, 76, 77</td></tr><tr><td>Bellet C.</td><td>60</td></tr><tr><td>Berny C.</td><td>99</td></tr><tr><td>Berry N. G.</td><td>51</td></tr><tr><td>Bertaux J.</td><td>53</td></tr><tr><td>Berthomieu A.</td><td>61</td></tr><tr><td>Bevins D.</td><td>89, 93</td></tr><tr><td>Bian G.</td><td>22, 93</td></tr><tr><td>Bleidorn C.</td><td>12, 78</td></tr><tr><td>Boivin T.</td><td>114</td></tr><tr><td>Bordenstein S.</td><td>44</td></tr><tr><td>Bouchon D.</td><td>52, 53, 72, 80, 109</td></tr><tr><td>Boucias D. G.</td><td>48</td></tr><tr><td>Bourret J.</td><td>92</td></tr><tr><td>Bourtzis K.</td><td>26, 27, 48, 116</td></tr><tr><td>Bouvaine S.</td><td>17</td></tr><tr><td>Braquart-Varnier C.</td><td>52, 109</td></tr><tr><td>Brelsfoard C. L.</td><td>20</td></tr><tr><td>Brucker R.</td><td>44</td></tr><tr><td>Bykov R.</td><td>15</td></tr></table>

-C-

<table><tr><td>Caceres C.</td><td>27</td></tr><tr><td>Cafiso A.</td><td>98</td></tr><tr><td>Calvitti M.</td><td>21, 100</td></tr><tr><td>Candau J. N.</td><td>114</td></tr><tr><td>Cárdenas J.</td><td>92</td></tr><tr><td>Cariou M.</td><td>10, 79</td></tr><tr><td>Carpentier M. C.</td><td>52</td></tr><tr><td>Cass B. N.</td><td>37</td></tr><tr><td>Cassidy A.</td><td>88</td></tr><tr><td>Charlat S.</td><td>10, 18, 79</td></tr><tr><td>Chen Y. N.</td><td>117</td></tr><tr><td>Chevalier F.</td><td>52, 109</td></tr><tr><td>Choe J. C.</td><td>16, 87, 102</td></tr><tr><td>Chouaia B.</td><td>98</td></tr></table>

<table><tr><td>Chrostek E.</td><td>64</td></tr><tr><td>Clare R.</td><td>88</td></tr><tr><td>Comandatore F.</td><td>63</td></tr><tr><td>Cook D. A.</td><td>88</td></tr><tr><td>Cook J. M.</td><td>54, 91</td></tr><tr><td>Cordaux R.</td><td>65, 66, 75</td></tr><tr><td>Couchoux C.</td><td>81</td></tr><tr><td>Crain P.</td><td>24</td></tr><tr><td>Curry M. M.</td><td>58</td></tr></table>

-D-

<table><tr><td>Darby A.</td><td>42</td></tr><tr><td>Davies J.</td><td>106</td></tr><tr><td>Daxböck-Horvath S.</td><td>104</td></tr><tr><td>de Castro Guimaraes A.</td><td>106</td></tr><tr><td>De Liberato C.</td><td>100</td></tr><tr><td>Deehan M.</td><td>30</td></tr><tr><td>Desiderio A.</td><td>21</td></tr><tr><td>Desouhant E.</td><td>99</td></tr><tr><td>Dittmer J.</td><td>80</td></tr><tr><td>Dobson S. L.</td><td>20, 24</td></tr><tr><td>Dodson B. L.</td><td>25</td></tr><tr><td>Doudoumis V.</td><td>27</td></tr><tr><td>Dumas E.</td><td>61</td></tr><tr><td>Duplouy A.</td><td>81</td></tr><tr><td>Duret L.</td><td>79</td></tr><tr><td>Duron O.</td><td>61</td></tr></table>

-E-

<table><tr><td>Egan S. P.</td><td>105</td></tr><tr><td>Ehrmann L.</td><td>34</td></tr><tr><td>Engl T.</td><td>56</td></tr><tr><td>Epis S.</td><td>98</td></tr><tr><td>Ernenwein L.</td><td>65</td></tr></table>

-F-

<table><tr><td>Fast E.</td><td>30, 59</td></tr><tr><td>Feder J. L.</td><td>105</td></tr><tr><td>Fiston-Lavier A. S.</td><td>61</td></tr><tr><td>Ford L.</td><td>51, 88, 106</td></tr><tr><td>Foster J. M.</td><td>72, 96</td></tr><tr><td>Francine T. N.</td><td>21</td></tr><tr><td>Freilich S.</td><td>38</td></tr><tr><td>Frydman H.</td><td>30, 31, 32, 59</td></tr><tr><td>Fukatsu T.</td><td>62, 71, 74, 82, 101</td></tr><tr><td>Furlong R. B.</td><td>95, 108</td></tr></table>

-G-

<table><tr><td>Gansauge M. T.</td><td>12</td></tr><tr><td>Gebiola M.</td><td>83, 110</td></tr><tr><td>Geniez S.</td><td>72</td></tr><tr><td>Genty L. M.</td><td>53</td></tr><tr><td>Gerth M.</td><td>12, 78</td></tr><tr><td>Ghosh S.</td><td>17</td></tr><tr><td>Gidoin C.</td><td>114</td></tr><tr><td>Gilbert C.</td><td>65</td></tr><tr><td>Gilles J.</td><td>116</td></tr><tr><td>Giorgini M.</td><td>83, 84</td></tr><tr><td>Giraud I.</td><td>65, 66, 75</td></tr><tr><td>Gottlieb Y.</td><td>40</td></tr><tr><td>Gowda M.</td><td>17</td></tr><tr><td>Grève P.</td><td>52, 65, 66, 72, 75, 109</td></tr></table>
<table><tbody><tr><td>Grobler Y.</td><td>33</td></tr><tr><td>Gruntenko N.</td><td>15</td></tr><tr><td>Grziwotz F.</td><td>24, 85</td></tr><tr><td>Guerrero R.</td><td>92</td></tr><tr><td>—H—</td><td></td></tr><tr><td>Hammerstein P.</td><td>43, 45, 113</td></tr><tr><td>Hanski I.</td><td>81</td></tr><tr><td>Harumoto T.</td><td>71, 74</td></tr><tr><td>Hattori M.</td><td>82</td></tr><tr><td>Henri H.</td><td>114</td></tr><tr><td>Herranz J.</td><td>77</td></tr><tr><td>Herrera P.</td><td>86</td></tr><tr><td>Himler A. G.</td><td>37</td></tr><tr><td>Hoffmeister T.</td><td>104</td></tr><tr><td>Hong X. Y.</td><td>69</td></tr><tr><td>Hornett E.</td><td>18</td></tr><tr><td>Hosokawa T.</td><td>82, 101</td></tr><tr><td>Hsiesh C.</td><td>24</td></tr><tr><td>Huang D. W.</td><td>70</td></tr><tr><td>Hughes G. L.</td><td>25</td></tr><tr><td>Hummel T.</td><td>49</td></tr><tr><td>Hunter M. S.</td><td>37, 83, 97, 110</td></tr><tr><td>Hurst G.</td><td>18</td></tr><tr><td>Husnik F.</td><td>42</td></tr><tr><td>Hypsa V.</td><td>42</td></tr><tr><td>—I—</td><td></td></tr><tr><td>Ignatenko O.</td><td>15</td></tr><tr><td>Ilinsky Y.</td><td>15</td></tr><tr><td>Isberg R. R.</td><td>36</td></tr><tr><td>Ishikawa Y.</td><td>112</td></tr><tr><td>—J—</td><td></td></tr><tr><td>Jeong G.</td><td>16, 87, 102</td></tr><tr><td>Jiggins F. M.</td><td>26</td></tr><tr><td>Johnston K. L.</td><td>51, 88</td></tr><tr><td>Joshi D.</td><td>22, 89, 93</td></tr><tr><td>Junker K.</td><td>92</td></tr><tr><td>—K—</td><td></td></tr><tr><td>Kageyama D.</td><td>54, 90</td></tr><tr><td>Kaltenpoth M.</td><td>56</td></tr><tr><td>Kamath A.</td><td>30, 31, 32</td></tr><tr><td>Karpova E.</td><td>15</td></tr><tr><td>Kaur R.</td><td>49</td></tr><tr><td>Kayukawa T.</td><td>112</td></tr><tr><td>Kelly S. E.</td><td>37, 83, 97, 110</td></tr><tr><td>Kemp D.</td><td>91</td></tr><tr><td>Kern P.</td><td>54, 91</td></tr><tr><td>Kim K. L. H.</td><td>60</td></tr><tr><td>Klasson L.</td><td>40</td></tr><tr><td>Kobayashi Y.</td><td>113</td></tr><tr><td>Koehler G.</td><td>76</td></tr><tr><td>Köppler K.</td><td>104</td></tr><tr><td>Kramer L. D.</td><td>25</td></tr><tr><td>Kremer N.</td><td>99</td></tr><tr><td>Krumböck S.</td><td>104</td></tr><tr><td>Kumar S.</td><td>72</td></tr><tr><td>Kyritsis G.</td><td>27</td></tr><tr><td>—L—</td><td></td></tr><tr><td>Lachowska-Cierlik D.</td><td>115</td></tr><tr><td>Lahav T.</td><td>38</td></tr><tr><td>Lalzar I.</td><td>40</td></tr><tr><td>Leclercq S.</td><td>65</td></tr><tr><td>Lefoulon E.</td><td>92</td></tr></tbody></table>

<table><tbody><tr><td>Lehmann R.</td><td>33</td></tr><tr><td>Lesobre J.</td><td>80</td></tr><tr><td>Lesser C. F.</td><td>36</td></tr><tr><td>Li J. J.</td><td>55, 117</td></tr><tr><td>Liang X.</td><td>22, 93</td></tr><tr><td>Lindsey A. R. I.</td><td>11</td></tr><tr><td>Linke B.</td><td>94</td></tr><tr><td>Liua C.</td><td>55</td></tr><tr><td>Longdon B.</td><td>26</td></tr><tr><td>LoVullo E. D.</td><td>95, 108</td></tr><tr><td>Lu P.</td><td>22</td></tr><tr><td>Luck A. N.</td><td>96</td></tr><tr><td>—M—</td><td></td></tr><tr><td>Mains J.</td><td>24</td></tr><tr><td>Mains J. W.</td><td>20</td></tr><tr><td>Makepeace B. L.</td><td>50</td></tr><tr><td>Makoundou P.</td><td>61</td></tr><tr><td>Malone C.</td><td>33</td></tr><tr><td>Mann E.</td><td>97</td></tr><tr><td>Mappa G.</td><td>109</td></tr><tr><td>Marini F.</td><td>21, 100</td></tr><tr><td>Martin C.</td><td>92</td></tr><tr><td>Martin E.</td><td>98</td></tr><tr><td>Martinez J.</td><td>26</td></tr><tr><td>Martínez-Rodríguez P.</td><td>13, 68, 76, 77</td></tr><tr><td>Mavingui P.</td><td>60</td></tr><tr><td>Mcfadden M.</td><td>89</td></tr><tr><td>McGraw B.</td><td>23</td></tr><tr><td>Mereghetti V.</td><td>98</td></tr><tr><td>Mialdea G.</td><td>10</td></tr><tr><td>Michalkova V.</td><td>56</td></tr><tr><td>Miki T.</td><td>24, 111</td></tr><tr><td>Miller W. J.</td><td>26, 27, 34, 48, 49, 56</td></tr><tr><td>Minard G.</td><td>60</td></tr><tr><td>Moné Y.</td><td>52</td></tr><tr><td>Monnin D.</td><td>99</td></tr><tr><td>Montagna M.</td><td>98</td></tr><tr><td>Moretti R.</td><td>21, 100</td></tr><tr><td>Moriyama M.</td><td>82, 101</td></tr><tr><td>Moumen B.</td><td>52, 65, 66, 72</td></tr><tr><td>Mozes-Daube N.</td><td>37</td></tr><tr><td>—N—</td><td></td></tr><tr><td>Nappo A. G.</td><td>84</td></tr><tr><td>Newton I. L. G.</td><td>36, 107</td></tr><tr><td>Nguyen D.</td><td>46</td></tr><tr><td>Nichols R. A.</td><td>68</td></tr><tr><td>Nikoh N.</td><td>74, 82, 101</td></tr><tr><td>Nixon G. L.</td><td>51</td></tr><tr><td>Noël C.</td><td>52</td></tr><tr><td>Noh P.</td><td>16, 87, 102</td></tr><tr><td>Novakova E.</td><td>42</td></tr><tr><td>—O—</td><td></td></tr><tr><td>Ok S.</td><td>26</td></tr><tr><td>O'Neill P. M.</td><td>51</td></tr><tr><td>O'Neill S.</td><td>23</td></tr><tr><td>Oshima K.</td><td>82</td></tr><tr><td>—P—</td><td></td></tr><tr><td>Pan X.</td><td>22</td></tr><tr><td>Papadopoulos N.</td><td>27</td></tr><tr><td>Park S.</td><td>16, 87, 102</td></tr><tr><td>Parker A. G.</td><td>48</td></tr><tr><td>Parrella G.</td><td>84</td></tr></tbody></table>
<table>
  <tbody>
    <tr>
      <td>Pastok D.</td>
      <td>18</td>
    </tr>
    <tr>
      <td>Pérez-Ruiz M.</td>
      <td>76, 77</td>
    </tr>
    <tr>
      <td>Pigeault R.</td>
      <td>109</td>
    </tr>
    <tr>
      <td>Pontier S. M.</td>
      <td>103</td>
    </tr>
    <tr>
      <td>—R—</td>
      <td></td>
    </tr>
    <tr>
      <td>Rabe A.</td>
      <td>35</td>
    </tr>
    <tr>
      <td>Raimond M.</td>
      <td>53</td>
    </tr>
    <tr>
      <td>Rasgon J. L.</td>
      <td>25</td>
    </tr>
    <tr>
      <td>Rasool B.</td>
      <td>104</td>
    </tr>
    <tr>
      <td>Reynolds L.</td>
      <td>18</td>
    </tr>
    <tr>
      <td>Riegler M.</td>
      <td>46, 54, 91, 104</td>
    </tr>
    <tr>
      <td>Rong X.</td>
      <td>69</td>
    </tr>
    <tr>
      <td>Roques A.</td>
      <td>114</td>
    </tr>
    <tr>
      <td>Rose R. I.</td>
      <td>20</td>
    </tr>
    <tr>
      <td>Rota-Stabelli O.</td>
      <td>49</td>
    </tr>
    <tr>
      <td>Rull J.</td>
      <td>105</td>
    </tr>
    <tr>
      <td>Ryder J.</td>
      <td>18</td>
    </tr>
    <tr>
      <td>—S—</td>
      <td></td>
    </tr>
    <tr>
      <td>Sagot M. F.</td>
      <td>10</td>
    </tr>
    <tr>
      <td>Schlick-Steiner B. C.</td>
      <td>94, 104</td>
    </tr>
    <tr>
      <td>Schmitz-Esser S.</td>
      <td>97</td>
    </tr>
    <tr>
      <td>Schneider D.</td>
      <td>27, 34, 48, 56</td>
    </tr>
    <tr>
      <td>Schuler H.</td>
      <td>104, 105</td>
    </tr>
    <tr>
      <td>Schwarz D.</td>
      <td>104</td>
    </tr>
    <tr>
      <td>Schweisguth F.</td>
      <td>103</td>
    </tr>
    <tr>
      <td>Serbus L.</td>
      <td>35</td>
    </tr>
    <tr>
      <td>Sharma R.</td>
      <td>106</td>
    </tr>
    <tr>
      <td>Sheehan K. B.</td>
      <td>36, 107</td>
    </tr>
    <tr>
      <td>Shinoda T.</td>
      <td>112</td>
    </tr>
    <tr>
      <td>Shirk P. D.</td>
      <td>95, 108</td>
    </tr>
    <tr>
      <td>Shone A.</td>
      <td>106</td>
    </tr>
    <tr>
      <td>Sicard M.</td>
      <td>61, 109</td>
    </tr>
    <tr>
      <td>Simhadri R. K.</td>
      <td>30, 31, 32, 59</td>
    </tr>
    <tr>
      <td>Simões P.</td>
      <td>10</td>
    </tr>
    <tr>
      <td>Slatko B. E.</td>
      <td>72, 96</td>
    </tr>
    <tr>
      <td>Sochova E.</td>
      <td>42</td>
    </tr>
    <tr>
      <td>Spooner-Hart R.</td>
      <td>46</td>
    </tr>
    <tr>
      <td>Stauffer C.</td>
      <td>104, 105</td>
    </tr>
    <tr>
      <td>Steiner F. M.</td>
      <td>94, 104</td>
    </tr>
    <tr>
      <td>Stouthamer C. M.</td>
      <td>97, 110</td>
    </tr>
    <tr>
      <td>Stouthamer R.</td>
      <td>11</td>
    </tr>
    <tr>
      <td>Strauss J. F.</td>
      <td>111</td>
    </tr>
    <tr>
      <td>Strunov A. A.</td>
      <td>34</td>
    </tr>
    <tr>
      <td>Sugihara G.</td>
      <td>24</td>
    </tr>
    <tr>
      <td>Sugimoto T. N.</td>
      <td>112</td>
    </tr>
    <tr>
      <td>Sullivan W.</td>
      <td>35</td>
    </tr>
    <tr>
      <td>Szöllösi G.</td>
      <td>10</td>
    </tr>
    <tr>
      <td>—T—</td>
      <td></td>
    </tr>
    <tr>
      <td>Tadeo E.</td>
      <td>105</td>
    </tr>
    <tr>
      <td>Tanaka K.</td>
      <td>74</td>
    </tr>
    <tr>
      <td>Tanya V. N.</td>
      <td>50</td>
    </tr>
    <tr>
      <td>Taylor M. J.</td>
      <td>51, 88, 106</td>
    </tr>
    <tr>
      <td>Teixeira L.</td>
      <td>26, 41, 64</td>
    </tr>
  </tbody>
</table>

<table>
  <tbody>
    <tr>
      <td>Telschow A.</td>
      <td>24, 45, 85, 111, 113</td>
    </tr>
    <tr>
      <td>Thézé J.</td>
      <td>65</td>
    </tr>
    <tr>
      <td>Thuy T. H. T.</td>
      <td>60</td>
    </tr>
    <tr>
      <td>Toomey M.</td>
      <td>30, 31, 32</td>
    </tr>
    <tr>
      <td>Tran F. H.</td>
      <td>60</td>
    </tr>
    <tr>
      <td>Truitt A. M.</td>
      <td>14</td>
    </tr>
    <tr>
      <td>Tsiamis G.</td>
      <td>27</td>
    </tr>
    <tr>
      <td>Tsuchida T.</td>
      <td>112</td>
    </tr>
    <tr>
      <td>Turner J. D.</td>
      <td>51, 106</td>
    </tr>
    <tr>
      <td>—U—</td>
      <td></td>
    </tr>
    <tr>
      <td>Unwin V. T.</td>
      <td>88</td>
    </tr>
    <tr>
      <td>—V—</td>
      <td></td>
    </tr>
    <tr>
      <td>Valiente Moro C.</td>
      <td>60</td>
    </tr>
    <tr>
      <td>van Nouhuys S.</td>
      <td>81</td>
    </tr>
    <tr>
      <td>Van V. T.</td>
      <td>60</td>
    </tr>
    <tr>
      <td>Vavre F.</td>
      <td>52, 75, 99, 114</td>
    </tr>
    <tr>
      <td>Veber P.</td>
      <td>114</td>
    </tr>
    <tr>
      <td>—W—</td>
      <td></td>
    </tr>
    <tr>
      <td>Waclawik B.</td>
      <td>115</td>
    </tr>
    <tr>
      <td>Wang G. H.</td>
      <td>70</td>
    </tr>
    <tr>
      <td>Wang J. L.</td>
      <td>117</td>
    </tr>
    <tr>
      <td>Wang Y. F.</td>
      <td>55, 117</td>
    </tr>
    <tr>
      <td>Wanga J. L.</td>
      <td>55</td>
    </tr>
    <tr>
      <td>Ward S. A.</td>
      <td>51, 88, 106</td>
    </tr>
    <tr>
      <td>Warzyszynska I.</td>
      <td>115</td>
    </tr>
    <tr>
      <td>Wastling J. M.</td>
      <td>50</td>
    </tr>
    <tr>
      <td>Waterhouse D.</td>
      <td>106</td>
    </tr>
    <tr>
      <td>Weigert A.</td>
      <td>12</td>
    </tr>
    <tr>
      <td>Weill M.</td>
      <td>61</td>
    </tr>
    <tr>
      <td>Weisman N.</td>
      <td>15</td>
    </tr>
    <tr>
      <td>Weiss B.</td>
      <td>56</td>
    </tr>
    <tr>
      <td>Werren J. H.</td>
      <td>113</td>
    </tr>
    <tr>
      <td>White J. A.</td>
      <td>58</td>
    </tr>
    <tr>
      <td>White P.</td>
      <td>35</td>
    </tr>
    <tr>
      <td>Won Y. J.</td>
      <td>102</td>
    </tr>
    <tr>
      <td>—X—</td>
      <td></td>
    </tr>
    <tr>
      <td>Xi Z.</td>
      <td>22, 89, 93, 116</td>
    </tr>
    <tr>
      <td>Xiao J. H.</td>
      <td>70</td>
    </tr>
    <tr>
      <td>Xiong E. J.</td>
      <td>55</td>
    </tr>
    <tr>
      <td>—Y—</td>
      <td></td>
    </tr>
    <tr>
      <td>Ye H.</td>
      <td>23</td>
    </tr>
    <tr>
      <td>Yu X. Q.</td>
      <td>55</td>
    </tr>
    <tr>
      <td>Yuan L. L.</td>
      <td>55</td>
    </tr>
    <tr>
      <td>Yudina M.</td>
      <td>15</td>
    </tr>
    <tr>
      <td>—Z—</td>
      <td></td>
    </tr>
    <tr>
      <td>Zakharenko L.</td>
      <td>15</td>
    </tr>
    <tr>
      <td>Zakharov I.</td>
      <td>15</td>
    </tr>
    <tr>
      <td>Zchori-Fein E.</td>
      <td>37, 38</td>
    </tr>
    <tr>
      <td>Zhang D.</td>
      <td>116</td>
    </tr>
    <tr>
      <td>Zhang F.</td>
      <td>89</td>
    </tr>
    <tr>
      <td>Zhang Y. K.</td>
      <td>69</td>
    </tr>
    <tr>
      <td>Zheng Y.</td>
      <td>55, 117</td>
    </tr>
    <tr>
      <td>Zug R.</td>
      <td>43</td>
    </tr>
  </tbody>
</table>
List of participants

Ghaith Aljayyoussi
Liverpool School of Tropical Medicine
Liverpool, UK
susie.crossman@liv.ac.uk

Hisashi Anbutsu
AIST
Tsukuba, Japan
h-anbutsu@aist.go.jp

Wolfgang Arthofer
University of Innsbruck
Innsbruck, Austria
wolfgang.arthofer@uibk.ac.at

Patrizia Arthofer-Peer
University of Innsbruck
Innsbruck, Austria
part@peerart.at

Myriam Badawi
Université de Poitiers
Poitiers, France
myriam.badawi@univ-poitiers.fr

José L. Bella
Universidad Autónoma de Madrid
Madrid, Spain
bella@uam.es

Joanne Bertaux
Université de Poitiers
Poitiers, France
joanne.bertaux@univ-poitiers.fr

Christoph Bleidorn
University of Leipzig
Leipzig, Germany
bleidorn@uni-leipzig.de

Seth Bordenstein
Vanderbilt University
Nashville, USA
s.bordenstein@vanderbilt.edu

Didier Bouchon
Université de Poitiers
Poitiers, France
didier.bouchon@univ-poitiers.fr

Kostas Bourtzis
International Atomic Energy Agency
Vienna, Austria
K.Bourtzis@iaea.org

Sophie Bouvaine
Natural Resources Institute
Chatham maritime, UK
bs33@gre.ac.uk

Alessandra Cafiso
Università degli Studi di Milano
Milano, Italia
alessandra.cafiso@unimi.it

Maurizio Calvitti
ENEA
Rome, Italy
maurizio.calvitti@enea.it

Marie Cariou
Université Lyon 1
Villeurbanne, France
marie.cariou@univ-lyon1.fr

Julien Cattel
Université Lyon 1
Villeurbanne Cedex, France
julien.cattel@univ-lyon1.fr

Sylvain Charlat
Université Lyon 1
Villeurbanne, France
sylvain.charlat@univ-lyon1.fr

Ewa Chrostek
Instituto Gulbenkian de Ciência
Oeiras, Portugal
echrostek@igc.gulbenkian.pt

Francesco Comandatore
University of Milan
Milano, Italia
francesco.comandatore@unimi.it

Richard Cordaux
Université de Poitiers
Poitiers, France
richard.cordaux@univ-poitiers.fr

Katherina Damisch
University of Innsbruck
Innsbruck, Austria
katherina.damisch@student.uibk.ac.at

Jessica Dittmer
Université de Poitiers
Poitiers, France
jessica.dittmer@univ-poitiers.fr
Stephen Dobson
University of Kentucky
Lexington, USA
sdobson@uky.edu

Anne Duplouy
Metapopulation Research Group
Helsinki, Finland
anne.duplouy@helsinki.fi

Sara Epis
Università degli Studi di Milano
Milano, Italia
sara.epis@guest.unimi.it

Louise Ford
Liverpool School of Tropical Medicine
Liverpool, UK
louise.ford@liverpool.ac.uk

Crystal Frost
University of Liverpool
Liverpool, UK
Crystal.Frost@liverpool.ac.uk

Horacio Frydman
Boston University
Boston, USA
hfrydman@bu.edu

Takema Fukatsu
AIST
Tsukuba, Japan
t-fukatsu@aist.go.jp

Marco Gebiola
The University of Arizona
Tucson, USA
marco.gebiola@gmail.com

Michael Gerth
Universität Leipzig
Leipzig, Germany
michael.gerth@uni-leipzig.de

Massimo Giorgini
Consiglio Nazionale delle Ricerche
Portici, Italy
giorgini@ipp.cnr.it

Yuval Gottlieb
Hebrew University
Rehovot, Israel
yuvalgd@yahoo.com

Pierre Greve
Université de Poitiers
Poitiers, France
pierre.greve@univ-poitiers.fr

Alexandra Grosbusch
University of Innsbruck
Innsbruck, Austria
alexandra.grosbusch@student.uibk.ac.at

Florian Grziwotz
Institute for Evolution and Biodiversity
Münster, Germany
f_grzi01@wwu.de

Sarina Hammerle
University of Innsbruck
Innsbruck, Austria
sarina.hammerle@student.uibk.ac.at

Peter Hammerstein
Humboldt University Berlin
Berlin, Germany
p.hammerstein@biologie.hu-berlin.de

Toshiyuki Harumoto
AIST
Tsukuba, Japan
t-harumoto@aist.go.jp

Paul Herrera
University of Veterinary Medicine
Wien, Vienna
paulherrera88@gmail.com

Xiao-Yue Hong
Nanjing Agricultural University
Nanjing, CHINA
xyhong@njau.edu.cn

Molly Hunter
The University of Arizona
Tucson, USA
mhunter@ag.arizona.edu

Greg Hurst
University of Liverpool
Liverpool, UK
g.hurst@liv.ac.uk

Filip Husnik
University of South Bohemia
Ceske Budejovice, Czech Republic
filip@paru.cas.cz

Yury Ilinsky
Siberian Branch of the Russian Academy of
Sciences
Novosibirsk, Russia
paulee@bionet.nsc.ru

Gilsang Jeong
Ewha Womans University
Seoul, Republic of Korea
ecodever@gmail.com
Kelly Johnston
Liverpool School of Tropical Medicine
Liverpool, UK
johnkel@liverpool.ac.uk

Deepak Joshi
Michigan State University
East Lansing, USA
djoshi@msu.edu

Daisuke Kageyama
National Institute of Agrobiological Sciences
Ibaraki, Japan
kagymad@affrc.go.jp

Martin Kaltenpoth
Max Planck Institute for Chemical Ecology
Jena, Germany
mkaltenpoth@ice.mpg.de

Ajit Kamath
Boston University
Boston, USA
ajit9988@bu.edu

Rupinder Kaur
Fondazione Edmund Mach
San Michele all'adige, Italy
rupinderkaur@fmach.it

Peter Kern
University of Western Sydney
Penrith, Australia
p.kern@uws.edu.au

Philipp Kirschner
University of Innsbruck
Innsbruck, Austria
philipp.kirschner@student.uibk.ac.at

Lisa Klasson
Uppsala University
Uppsala, Sweden
lisa.klasson@icm.uu.se

Natacha Kremer
Université Lyon 1
Villeurbanne, France
natacha.kremer@normalesup.org

Emilie Lefoulon
Muséum National d'Histoire Naturelle
Paris, France
elefoulon@mnhn.fr

Tera Levin
University of California, Berkeley
Berkeley, USA
teralevin@gmail.com

Xiao Liang
Michigan State University
East Lansing, USA
liangxiao0611@gmail.com

Amelia Lindsey
University of California Riverside
Riverside, USA
alind005@ucr.edu

Bernd Linke
University of Innsbruck
Innsbruck, Austria
bernd.linke@student.uibk.ac.at

Eric LoVullo
United States Department of Agriculture
Gainesville, USA
eric.lovullo@ars.usda.gov

Ashley Luck
New England Biolabs
Ipswich, USA
luck@neb.com

Benjamin Makepeace
University of Liverpool
Liverpool, UK
blm1@liv.ac.uk

Colin Malone
NYUMC - Skirball Institute
New York, USA
colin.malone@med.nyu.edu

Evelyne Mann
University of Veterinary Medicine
Vienna, Austria
evelyne.mann@vetmeduni.ac.at

Elena Martin
Università dagli Studi di Milano
Milano, Italia
elena.martin@unimi.it

Julien Martinez
University of Cambridge
Cambridge, UK
jtm35@cam.ac.uk

Paloma Martínez-Rodríguez
Universidad Autónoma de Madrid
Madrid, Spain
paloma.martinez@uam.es

Elizabeth McGraw
Monash University
Melbourne, Australia
beth.mcgraw@monash.edu
Wolfgang Miller
Medical University of Vienna
Vienna, Austria
wolfgang.miller@meduniwien.ac.at

David Monnin
Université Lyon 1
Villeurbanne, France
david.monnin@univ-lyon1.fr

Riccardo Moretti
ENEA
Rome, Italy
riccardo.moretti@enea.it

Minoru Moriyama
AIST
Tusukuba, Japan
m-moriyama@aist.go.jp

Bouziane Moumen
Université de Poitiers
Poitiers, France
bouziane.moumen@univ-poitiers.fr

Irene Newton
Indiana University
Bloomington, USA
irnewton@indiana.edu

Richard Nichols
Queen Mary University of London
Londón, UK
r.a.nichols@qmul.ac.uk

Soyeon Park
Ewha Womans University
Seoul, Republic of Korea
sy.soyeon@gmail.com

Caroline Pichler
University of Innsbruck
Innsbruck, Austria
caroline.pichler@uibk.ac.at

Stephanie Pontier
Institut Pasteur
Paris, France
stephanie.pontier@pasteur.fr

Jason Rasgon
Penn State University
State College, USA
jlr54@psu.edu

Markus Riegler
University of Western Sydney
Penrith, Australia
m.riegler@uws.edu.au

Heike Ritthammer
University of Innsbruck
Innsbruck, Austria
heike.ritthammer@student.uibk.ac.at

Sarah Scheld
University of Innsbruck
Innsbruck, Austria
sarah.scheld@student.uibk.ac.at

Birgit C. Schlick-Steiner
University of Innsbruck
Innsbruck, Austria
birgit.schlick-steiner@uibk.ac.at

Stephan Schmitz-Esser
University of Veterinary Medicine
Vienna, Austria
stephan.schmitz-esser@vetmeduni.ac.at

Daniela Schneider
Medical University of Vienna
Vienna, Austria
daniela.schneider@meduniwien.ac.at

Hannes Schuler
Free University of Bozen-Bolzano
Bozen, Italy
hannes.schuler@boku.ac.at

Julia Seeber
University of Innsbruck
Innsbruck, Austria
julia.seeber@uibk.ac.at

Laura Serbus
Florida International University
Miami, USA
lserbus@fiu.edu

Raman Sharma
Liverpool School of Tropical Medicine
Liverpool, UK
rsharma8@liv.ac.uk

Kathy Sheehan
Indiana University
Bloomington, USA
sheehaka@gmail.com

Paul Shirk
United States Department of Agriculture
Gainesville, USA
paul.shirk@ars.usda.gov

Mathieu Sicard
Université Montpellier 2
Montpellier, France
Mathieu.sicard@univ-montp2.fr
Rama Krishna Simhadri
Boston University
Boston, USA
ramaks@bu.edu

Barton Slatko
New England Biolabs
Ipswich, USA
slatko@neb.com

Christian Stauffer
Institute of Forest Entomology
Vienna, Austria
christian.stauffer@boku.ac.at

Florian M. Steiner
University of Innsbruck
Innsbruck, Austria
florian.m.steiner@uibk.ac.at

Iris S. Steiner
University of Innsbruck
Innsbruck, Austria
irsteiner@tsn.at

Julia S. Steiner
University of Innsbruck
Innsbruck, Austria
jsteiner1@tsn.at

Corinne Stouthamer
University of Arizona
Tucson, USA
cmstouthamer@gmail.com

Jakob Friedrich Strauss
Westfälische Wilhelms Universitaet Muenster
Muenster, Germany
jfstrauss@gmail.com

Takafumi Sugimoto
University of Toyama
Toyama, Japan
sugimoto@ctg.u-toyama.ac.jp

Mark Taylor
Liverpool School of Tropical Medicine
Liverpool, UK
susie.crossman@liv.ac.uk

Luís Teixeira
Instituto Gulbenkian de Ciência
Oeiras, Portugal
lteixeira@igc.gulbenkian.pt

Arndt Telschow
Westfalian Wilhelsm-University Münster
Münster, Germany
a.telschow@wwu.de

Michelle Toomey
Boston University
Woburn, USA
mtoomey5@bu.edu

Amy Truitt
Portland State University
Portland, USA
amtruitt@pdx.edu

Claire Valiente Moro
Université Lyon 1
Villeurbanne, France
claire.valiente-moro@univ-lyon1.fr

Fabrice Vavre
Université Lyon 1
Villeurbanne, France
fabrice.vavre@univ-lyon1.fr

Beniamin Waclawik
Jagiellonian University
Krakow, Poland
beniamin.waclawik@uj.edu.pl

Herbert C. Wagner
University of Innsbruck
Innsbruck, Austria
herbert.wagner@uibk.ac.at

Guan-Hong Wang
Chinese Academy of Sciences
Beijing, China
wangguanhong@ioz.ac.cn

Yu-Feng Wang
Central China Normal University
Wuhan, China
yfengw@mail.ccnu.edu.cn

Mylène Weill
Université Montpellier 2
Montpellier, France
mylene.weill@univ-montp2.fr

Jen White
University of Kentucky
Lexington, USA
jenawhite@uky.edu

Zhiyong Xi
Michigan State University
East Lansing, USA
xizy@msu.edu

Einat Zchori-Fein
Newe Ya'ar Research Center
Ramat Yishay, Israel
einat@agri.gov.il
Dongjing Zhang
International Atomic Energy Agency
Vienna, Austria
zhangdongjing06@163.com

Ya Zheng
Central China Normal University
Wuhan, China
zhengyahb@163.com

Roman Zug
Humboldt-Universität zu Berlin
Berlin, Germany
roman.zug@biologie.hu-berlin.de
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