Suzanne E. Kelly

Rapid Spread of a Bacterial Symbiont in an Invasive Whitefly Is Driven by Fitness Benefits and Female Bias

A Rickettsia bacterium promotes its own geographical spread by manipulating its insect host’s sex ratio and fecundity. Maternally inherited bacterial symbionts of arthropods are common, yet symbiont invasions of host populations have rarely been observed. Here, we show that Rickettsia sp. nr. bellii swept into a population of an invasive agricultural pest, the sweet potato whitefly, Bemisia tabaci, in just 6 years. Compared with uninfected whiteflies, Rickettsia-infected whiteflies produced more offspring, had higher survival to adulthood, developed faster, and produced a higher proportion of daughters. The symbiont thus functions as both mutualist and reproductive manipulator. The observed increased performance and sex-ratio bias of infected whiteflies are sufficient to explain the spread of Rickettsia across the southwestern United States. Symbiont invasions such as this represent a sudden evolutionary shift for the host, with potentially large impacts on its ecology and invasiveness.

A newly discovered bacterium associated with parthenogenesis and a change in host selection behavior in parasitoid wasps.

The symbiotic bacterium Wolbachia pipientis has been considered unique in its ability to cause multiple reproductive anomalies in its arthropod hosts. Here we report that an undescribed bacterium is vertically transmitted and associated with thelytokous parthenogenetic reproduction in Encarsia, a genus of parasitoid wasps. Although Wolbachia was found in only one of seven parthenogenetic Encarsia populations examined, the “Encarsia bacterium” (EB) was found in the other six. Among seven sexually reproducing populations screened, EB was present in one, and none harbored Wolbachia. Antibiotic treatment did not induce male production in Encarsia pergandiella but changed the oviposition behavior of females. Cured females accepted one host type at the same rate as control females but parasitized significantly fewer of the other host type. Phylogenetic analysis based on the 16S rDNA gene sequence places the EB in a unique clade within the Cytophaga-Flexibacter-Bacteroid group and shows EB is unrelated to the Proteobacteria, where Wolbachia and most other insect symbionts are found. These results imply evolution of the induction of parthenogenesis in a lineage other than Wolbachia. Importantly, these results also suggest that EB may modify the behavior of its wasp carrier in a way that enhances its transmission.

A bacterial symbiont in the Bacteroidetes induces cytoplasmic incompatibility in the parasitoid wasp Encarsia pergandiella

Vertically transmitted symbionts of arthropods have been implicated in several reproductive manipulations of their hosts. These include cytoplasmic incompatibility (CI), parthenogenesis induction in haplodiploid species (PI), feminization and male killing. One symbiont lineage in the α–Proteobacteria, Wolbachia, is the only bacterium known to cause all of these effects, and has been thought to be unique in causing CI, in which the fecundity of uninfected females is reduced after mating with infected males. Here, we provide evidence that an undescribed symbiont in the Bacteroidetes group causes CI in a sexual population of the parasitic wasp Encarsia pergandiella. Wasps were crossed in all four possible combinations of infected and uninfected individuals. In the cross predicted to be incompatible, infected (I) males × MetaPress ×uninfected (U) females, progeny production was severely reduced, with these females producing only 12.6% of the number of progeny in other crosses. The incompatibility observed in this haplodiploid species was the female mortality type; dissections showed that most progeny from the incompatible cross died as eggs. The 16S rDNA sequence of this symbiont is 99% identical to a parthenogenesis-inducing symbiont in other Encarsia, and 96% identical to a feminizing symbiont in haplodiploid Brevipalpus mites. Thus , this recently discovered symbiont lineage is capable of inducing three of the four principal manipulations of host reproduction known to be caused by Wolbachia.

Almost There: Transmission Routes of Bacterial Symbionts between Trophic Levels

Many intracellular microbial symbionts of arthropods are strictly vertically transmitted and manipulate their hosts reproduction in ways that enhance their own transmission. Rare horizontal transmission events are nonetheless necessary for symbiont spread to novel host lineages. Horizontal transmission has been mostly inferred from phylogenetic studies but the mechanisms of spread are still largely a mystery. Here, we investigated transmission of two distantly related bacterial symbionts – Rickettsia and Hamiltonella – from their host, the sweet potato whitefly, Bemisia tabaci, to three species of whitefly parasitoids: Eretmocerus emiratus, Eretmocerus eremicus and Encarsia pergandiella. We also examined the potential for vertical transmission of these whitefly symbionts between parasitoid generations. Using florescence in situ hybridization (FISH) and transmission electron microscopy we found that Rickettsia invades Eretmocerus larvae during development in a Rickettsia-infected host, persists in adults and in females, reaches the ovaries. However, Rickettsia does not appear to penetrate the oocytes, but instead is localized in the follicular epithelial cells only. Consequently, Rickettsia is not vertically transmitted in Eretmocerus wasps, a result supported by diagnostic polymerase chain reaction (PCR). In contrast, Rickettsia proved to be merely transient in the digestive tract of Encarsia and was excreted with the meconia before wasp pupation. Adults of all three parasitoid species frequently acquired Rickettsia via contact with infected whiteflies, most likely by feeding on the host hemolymph (host feeding), but the rate of infection declined sharply within a few days of wasps being removed from infected whiteflies. In contrast with Rickettsia, Hamiltonella did not establish in any of the parasitoids tested, and none of the parasitoids acquired Hamiltonella by host feeding. This study demonstrates potential routes and barriers to horizontal transmission of symbionts across trophic levels. The possible mechanisms that lead to the differences in transmission of species of symbionts among species of hosts are discussed.

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.

Hyperparasitism by an exotic autoparasitoid: secondary host selection and the window of vulnerability of conspecific and native heterospecific hosts

Encarsia transvena is an ‘autoparasitoid’ in the hymenopteran family Aphelinidae. In this species, female eggs are laid in whitefly nymphs. Male eggs are laid externally on immature parasitoids enclosed within the whitefly integument, either their own species, or other primary parasitoids. We explored parasitism by E. transvena of conspecific female immatures and those of a native primary parasitoid, Eretmocerus eremicus, in laboratory experiments. In the first experiment, female E. transvena were offered different combinations of two stages of E. transvena (late larvae – prepupae (ET2), and early pupae (ET3)), and one stage of E. eremicus (prepupae – early pupae (EE2)) in paired choice tests. The results indicated very little parasitism of ET3 relative to the host it was paired with, either EE2 or ET2. However, when EE2 was offered with ET2, there was no statistically significant difference in parasitism. In a no‐choice experiment in which oviposition patterns and male progeny development were examined in four stages of both species of wasp, clear differences were observed between the host species. Only one stage of E. transvena (ET2) was parasitized and supported development of male E. transvena to any significant degree. In contrast, in E. eremicus, EE2, EE3 (red‐eyed pupae), and EE4 (late pupae) were all parasitized, and male E. transvena emerged from all three stages, although fewer males emerged from EE4. In both species, wasp larvae that were still enclosed within the wet whitefly remains (ET1 and EE1) were parasitized at a very low rate. Lastly, an experiment that determined the length of the later developmental stages of E. transvena and E. eremicus suggested that the duration of the period in which E. transvena is susceptible to parasitism by conspecific females is less than half the period of susceptibility of E. eremicus. These results taken together suggest the potential for interference of E. eremicus by E. transvena, but other factors not examined here may also influence the outcome of interactions in the field.

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.

DOES AN AUTOPARASITOID DISRUPT HOST SUPPRESSION PROVIDED BY A PRIMARY PARASITOID

Theory predicts that intraguild consumers such as predators or parasitoids may displace more specialized heterospecific competitors and thereby actually increase the population densities of a shared host or prey. We tested this idea with a native primary parasitoid, Eretmocerus eremicus, and an exotic autoparasitoid Encarsia sophia, both at- tacking the sweetpotato whitefly Bemisia tabaci. Autoparasitoids are intraguild consumers that attack and kill both the immature stages of hemipteran hosts, such as whiteflies, and heterospecific and conspecific parasitoids. In population cages on cotton plants in the field in 1997 and 1998, we introduced whiteflies and then parasitoids in a replacement design with constant total numbers of parasitoids, as follows: (1) control (whiteflies only), (2) E. eremicus only, (3) E. sophia only, and (4) both E. eremicus and E. sophia. Destructive samples of plants were taken 2, 4, and 6 wk after wasp releases, and immature whitefly and wasp stages were censused. In 1997, there were no significant differences in whitefly densities among treatments. In 1998, the control treatment densities were significantly greater than parasitoid treatments, but there was no difference among the parasitoid treatments, indicating equivalent sup- pression of whitefly populations by the two parasitoid species. In both years, similar patterns in parasitoid dynamics were observed. Densities of E. eremicus were significantly higher in the absence of E. sophia. In contrast, E. sophia densities were unaffected by the presence of E. eremicus. The results suggest that interference by the autoparasitoid reduced primary parasitoid density, but with no concommitant disruption of host suppression. The results support theoretical predictions that no disruption should occur when both parasitoids are equally efficient and suggest that an autoparasitoid may be as efficient as a primary par- asitoid in suppressing host densities.

Heterospecific ovicide influences the outcome of competition between two endoparasitoids, Encarsia formosa and Encarsia luteola

Abstract 1. Studies of inter‐specific competition in parasitoids have largely focused on the outcome of within‐host competition and the behavioural mechanisms by which female parasitoids prevent competition. Another, less well studied, possibility is oviposition preceded by ‘heterospecific ovicide’, the destruction of the other species’ egg. Heterospecific ovicide essentially eliminates within‐host competition.