Steve J. Perlman

Adaptation via Symbiosis: Recent Spread of a Drosophila Defensive Symbiont

Offsetting the Cost of Parasitism Fruit flies, like most animals, are vulnerable to infection by a range of organisms, which, in co-infections, can interact with sometimes surprising effects. Jaenike et al. (p. 212) discovered that a species of Spiroplasma bacterium that is sometimes found in flies, and that is transmitted from mother to offspring, protects its host from the effects of a nematode worm parasite, Howardula aoronymphium. The worm sterilizes the female flies and shortens their lives, but when flies were experimentally infected with Spiroplasma, their fertility was rescued. Similarly, in wild populations of fruit flies infected with worms, those also infected with Spiroplasma had more eggs in their ovaries. The bacterium inhibits the growth of the adult female worms, but such is the advantage of this bacterial infection in offsetting the burden of nematodes on reproductive fitness, Spiroplasma appears to be spreading rapidly through populations of fruit flies in North America. A bacterium protects fruit flies against a sterilizing worm parasite. Recent studies have shown that some plants and animals harbor microbial symbionts that protect them against natural enemies. Here we demonstrate that a maternally transmitted bacterium, Spiroplasma, protects Drosophila neotestacea against the sterilizing effects of a parasitic nematode, both in the laboratory and the field. This nematode parasitizes D. neotestacea at high frequencies in natural populations, and, until recently, almost all infections resulted in complete sterility. Several lines of evidence suggest that Spiroplasma is spreading in North American populations of D. neotestacea and that a major adaptive change to a symbiont-based mode of defense is under way. These findings demonstrate the profound and potentially rapid effects of defensive symbionts, which are increasingly recognized as major players in the ecology of species interactions.

Cytoplasmic incompatibility in the parasitic wasp Encarsia inaron: disentangling the roles of Cardinium and Wolbachia symbionts.

Many bacterial endosymbionts of insects are capable of manipulating their hosts reproduction for their own benefit. The most common strategy of manipulation is cytoplasmic incompatibility (CI), in which embryonic mortality results from matings between uninfected females and infected males. In contrast, embryos develop normally in infected females, whether or not their mate is infected, and infected progeny are produced. In this way, the proportion of infected females increases in the insect population, thereby promoting the spread of the maternally inherited bacteria. However, what happens when multiple endosymbionts inhabit the same host? The parasitoid wasp Encarsia inaron is naturally infected with two unrelated endosymbionts, Cardinium and Wolbachia, both of which have been documented to cause CI in other insects. Doubly infected wasps show the CI phenotype. We differentially cured E. inaron of each endosymbiont, and crossed hosts of different infection status to determine whether either or both bacteria caused the observed CI phenotype in this parasitoid, and whether the two symbionts interacted within their common host. We found that Wolbachia caused CI in E. inaron, but Cardinium did not. We did not find evidence that Cardinium was able to modify or rescue Wolbachia-induced CI, nor did we find that Cardinium caused progeny sex ratio distortion, leaving the role of Cardinium in E. inaron a mystery.

Population biology of cytoplasmic incompatibility: maintenance and spread of Cardinium symbionts in a parasitic wasp

Bacteria that cause cytoplasmic incompatibility (CI) are perhaps the most widespread parasites of arthropods. CI symbionts cause reproductive failure when infected males mate with females that are either uninfected or infected with a different, incompatible strain. Until recently, CI was known to be caused only by the α-proteobacterium Wolbachia. Here we present the first study of the population biology of Cardinium, a recently discovered symbiont in the Bacteroidetes that causes CI in the parasitic wasp Encarsia pergandiella (Hymenoptera: Aphelinidae). Cardinium occurs at high frequency (∼92%) in the field. Using wasps that were recently collected in the field, we measured parameters that are crucial for understanding how CI spreads and is maintained in its host. CI Cardinium exhibits near-perfect rates of maternal transmission, causes a strong reduction in viable offspring in incompatible crosses, and induces a high fecundity cost, with infected females producing 18% fewer offspring in the first 4 days of reproduction. We found no evidence for paternal transmission or horizontal transmission of CI Cardinium through parasitism of an infected conspecific. No evidence for cryptic parthenogenesis in infected females was found, nor was sex allocation influenced by infection. We incorporated our laboratory estimates into a model of CI dynamics. The model predicts a high stable equilibrium, similar to what we observed in the field. Interestingly, our model also predicts a high threshold frequency of CI invasion (20% for males and 24% for females), below which the infection is expected to be lost. We consider how this threshold may be overcome, focusing in particular on the sensitivity of CI models to fecundity costs. Overall our results suggest that the factors governing the dynamics of CI Wolbachia and Cardinium are strikingly similar.

Rickettsia felis Infection in a Common Household Insect Pest, Liposcelis bostrychophila (Psocoptera: Liposcelidae)

ABSTRACT Many species of Rickettsia are well-known mammalian pathogens transmitted by blood-feeding arthropods. However, molecular surveys are continually uncovering novel Rickettsia species, often in unexpected hosts, including many arthropods that do not feed on blood. This study reports a systematic molecular characterization of a Rickettsia infecting the psocid Liposcelis bostrychophila (Psocoptera: Liposcelidae), a common and cosmopolitan household pest. Surprisingly, the psocid Rickettsia is shown to be Rickettsia felis, a human pathogen transmitted by fleas that causes serious morbidity and occasional mortality. The plasmid from the psocid R. felis was sequenced and was found to be virtually identical to the one in R. felis from fleas. As Liposcelis insects are often intimately associated with humans and other vertebrates, it is speculated that they acquired R. felis from fleas. Whether the R. felis in psocids causes disease in vertebrates is not known and warrants further study.

Host defense via symbiosis in Drosophila.

Host-associated microbes have often been studied as pathogens and the causes of disease, but symbiotic microbes that benefit their hosts are now known to be ubiquitous. In particular, insects possess a diversity of bacteria that can defend against natural enemies—Anopheles mosquitoes, for example, were recently shown to host a gut bacterium that confers refractoriness to malaria parasites [1]. In Drosophila, a key model of infection and immunity, fascinating examples of defense are accumulating, and two lineages of bacteria that infect the genus are now known to be defensive: Wolbachia and Spiroplasma (Figure 1). Both are vertically transmitted, both are facultative in Drosophila in that they are not strictly required by the host, and both infect Drosophila melanogaster. Here, we summarize what is known of Drosophila as an intriguing and emerging model of defensive symbiosis. Drosophila is an incredibly diverse genus with thousands of species, many of which are infected by Wolbachia and Spiroplasma [2]. As maternally transmitted symbionts, Wolbachia and Spiroplasma came to attention in Drosophila through their ability to manipulate host reproduction to favor their own transmission. Wolbachia are notorious for doing this by inducing cytoplasmic incompatibility (CI), whereby matings between Wolbachia-infected males and uninfected females result in the production of few to no offspring [3], providing selective pressure to maintain and rapidly spread Wolbachia in host populations. Though Spiroplasma are not known to induce CI, both Spiroplasma and Wolbachia can selfishly distort host sex ratios through male-killing in Drosophila, selectively killing the male offspring of infected females [3,4]. Many strains of Wolbachia and Spiroplasma, though, do not have such manipulative tendencies, and it has largely been a mystery how they are maintained in host populations. The discovery that they can defend against enemies has gone a long way in explaining their persistence, and has begun to shift our perception of many facultative inherited symbionts from that of manipulative parasites toward helpful mutualists.

The Role of Host Phylogeny Varies in Shaping Microbial Diversity in the Hindguts of Lower Termites

ABSTRACT The hindguts of lower termites and Cryptocercus cockroaches are home to a distinct community of archaea, bacteria, and protists (primarily parabasalids and some oxymonads). Within a host species, the composition of these hindgut communities appears relatively stable, but the evolutionary and ecological factors structuring community composition and stability are poorly understood, as are differential impacts of these factors on protists, bacteria, and archaea. We analyzed the microbial composition of parabasalids and bacteria in the hindguts of Cryptocercus punctulatus and 23 species spanning 4 families of lower termites by pyrosequencing variable regions of the small-subunit rRNA gene. Especially for the parabasalids, these data revealed undiscovered taxa and provided a phylogenetic basis for a more accurate understanding of diversity, diversification, and community composition. The composition of the parabasalid communities was found to be strongly structured by the phylogeny of their hosts, indicating the importance of historical effects, although exceptions were also identified. Particularly, spirotrichonymphids and trichonymphids likely were transferred between host lineages. In contrast, host phylogeny was not sufficient to explain the majority of bacterial community composition, but the compositions of the Bacteroidetes, Elusimicrobia, Tenericutes, Spirochaetes, and Synergistes were structured by host phylogeny perhaps due to their symbiotic associations with protists. All together, historical effects probably resulting from vertical inheritance have had a prominent role in structuring the hindgut communities, especially of the parabasalids, but dispersal and environmental acquisition have played a larger role in community composition than previously expected.

Endosymbiont costs and benefits in a parasitoid infected with both Wolbachia and Cardinium

Theory suggests that maternally inherited endosymbionts can promote their spread and persistence in host populations by enhancing the production of daughters by infected hosts, either by improving overall host fitness, or through reproductive manipulation. In the doubly infected parasitoid wasp Encarsia inaron, Wolbachia manipulates host reproduction through cytoplasmic incompatibility (CI), but Cardinium does not. We investigated the fitness costs and/or benefits of infection by each bacterium in differentially cured E. inaron as a potential explanation for persistence of Cardinium in this population. We introgressed lines infected with Wolbachia, Cardinium or both with the cured line to create a similar genetic background, and evaluated several parasitoid fitness parameters. We found that symbiont infection resulted in both fitness costs and benefits for E. inaron. The cost was lower initial egg load for all infected wasps. The benefit was increased survivorship, which in turn increased male production for wasps infected with only Cardinium. Female production was unaffected by symbiont infection; we therefore have not yet identified a causal fitness effect that can explain the persistence of Cardinium in the population. Interestingly, the Cardinium survivorship benefit was not evident when Wolbachia was also present in the host, and the reproduction of doubly infected individuals did not differ significantly from uninfected wasps. Therefore, the results of our study show that even when multiple infections seem to have no effect on a host, there may be a complex interaction of costs and benefits among symbionts.

Ecology and Evolution of Host‐Parasite Associations: Mycophagous Drosophila and Their Parasitic Nematodes

Associations between mycophagous Drosophila and nematode parasites occur throughout the temperate and boreal regions of North America, Europe, and Asia. The nematode Howardula aoronymphium has substantial adverse effects on host survival and fertility on North American Drosophila. Long‐term data show that rainy summers lead to a high prevalence of parasitism in the fall and the following spring, resulting in up to a 1‐yr time lag between present rainfall and increased prevalence of H. aoronymphium parasitism. A biogeographic analysis of the relative abundance of different Drosophila species has shown that H. aoronymphium may facilitate the coexistence of different species of Drosophila that compete for larval food resources. The actual host range of parasites in nature is determined by the intrinsic suitability of potential hosts for parasite infection and reproduction and various ecological factors. For H. aoronymphium in eastern North America, intrinsically suitable hosts fall within a restricted clade within the genus Drosophila. However, the temperature sensitivity of H. aoronymphium prevents it from using several host species that occur outside the geographical range of the nematodes. Finally, the host range, virulence, and geographical range of Drosophila‐parasitic nematodes appear to be highly dynamic over evolutionary timescales.

A ribosome-inactivating protein in a Drosophila defensive symbiont

Significance Symbioses between animals and microbes are now recognized as critical to many aspects of host health. This is especially true in insects, which are associated with diverse maternally transmitted endosymbionts that can protect against parasites and pathogens. Here, we find that Spiroplasma—a defensive endosymbiont that protects Drosophila during parasitism by a virulent and common nematode—encodes a protein toxin, a ribosome-inactivating protein (RIP) related to bacterial virulence factors such as the Shiga-like toxins in Escherichia coli. We further find that nematode ribosomal RNA suffers depurination consistent with attack by a RIP when the host is protected by Spiroplasma, suggesting a mechanism through which symbiotic microbes may protect their hosts from disease. Vertically transmitted symbionts that protect their hosts against parasites and pathogens are well known from insects, yet the underlying mechanisms of symbiont-mediated defense are largely unclear. A striking example of an ecologically important defensive symbiosis involves the woodland fly Drosophila neotestacea, which is protected by the bacterial endosymbiont Spiroplasma when parasitized by the nematode Howardula aoronymphium. The benefit of this defense strategy has led to the rapid spread of Spiroplasma throughout the range of D. neotestacea, although the molecular basis for this protection has been unresolved. Here, we show that Spiroplasma encodes a ribosome-inactivating protein (RIP) related to Shiga-like toxins from enterohemorrhagic Escherichia coli and that Howardula ribosomal RNA (rRNA) is depurinated during Spiroplasma-mediated protection of D. neotestacea. First, we show that recombinant Spiroplasma RIP catalyzes depurination of 28S rRNAs in a cell-free assay, as well as Howardula rRNA in vitro at the canonical RIP target site within the α-sarcin/ricin loop (SRL) of 28S rRNA. We then show that Howardula parasites in Spiroplasma-infected flies show a strong signal of rRNA depurination consistent with RIP-dependent modification and large decreases in the proportion of 28S rRNA intact at the α-sarcin/ricin loop. Notably, host 28S rRNA is largely unaffected, suggesting targeted specificity. Collectively, our study identifies a novel RIP in an insect defensive symbiont and suggests an underlying RIP-dependent mechanism in Spiroplasma-mediated defense.

Transcriptional responses in a Drosophila defensive symbiosis

Inherited symbionts are ubiquitous in insects and can have important consequences for the fitness of their hosts. Many inherited symbionts defend their hosts against parasites or other natural enemies; however, the means by which most symbionts confer protection is virtually unknown. We examine the mechanisms of defence in a recently discovered case of symbiont‐mediated protection, where the bacterial symbiont Spiroplasma defends the fly Drosophila neotestacea from a virulent nematode parasite, Howardula aoronymphium. Using quantitative PCR of Spiroplasma infection intensities and whole transcriptome sequencing, we attempt to distinguish between the following modes of defence: symbiont–parasite competition, host immune priming and the production of toxic factors by Spiroplasma. Our findings do not support a model of exploitative competition between Howardula and Spiroplasma to mediate defence, nor do we find strong support for host immune priming during Spiroplasma infection. Interestingly, we recovered sequence for putative toxins encoded by Spiroplasma, including a novel putative ribosome‐inactivating protein, transcripts of which are up‐regulated in response to nematode exposure. Protection via the production of toxins may be a widely used and important mechanism in heritable defensive symbioses in insects.