Unexpected diversity of Wolbachia associated with Bactrocera dorsalis (Diptera: Tephritidae) in Africa

Bactrocera dorsalis (Hendel) is an important pest of fruit-bearing plants in many countries worldwide. In Africa, this pest has spread rapidly and has become widely established since the first invasion report in 2003. Wolbachia is a vertically transmitted endosymbiont that can significantly influence aspects of the biology and, in particular, the reproduction of its host. In this study, we screened B. dorsalis specimens collected from several locations in Africa between 2005 and 2017 for Wolbachia using a PCR-based assay to target the Wolbachia surface protein wsp. Of the 357 individuals tested, 10 were positive for Wolbachia using the wsp assay. We identified four strains of Wolbachia infecting two B. dorsalis mitochondrial haplotypes. We found no strict association between the infecting strain and host haplotype, with one strain being present in two different host haplotypes. All the detected strains belonged to Super Group B Wolbachia and did not match any strains reported previously in B. dorsalis in Asia. These findings indicate that diverse Wolbachia infections are present in invasive populations of B. dorsalis

Vertical transmission of a Drosophila endosymbiont via cooption of the yolk transport and internalization machinery

Spiroplasma is a diverse bacterial clade that includes many vertically transmitted insect endosymbionts, including Spiroplasma poulsonii, a natural endosymbiont of Drosophila melanogaster. These bacteria persist in the hemolymph of their adult host and exhibit efficient vertical transmission from mother to offspring. In this study, we analyzed the mechanism that underlies their vertical transmission, and here we provide strong evidence that these bacteria use the yolk uptake machinery to colonize the germ line. We show that Spiroplasma reaches the oocyte by passing through the intercellular space surrounding the ovarian follicle cells and is then endocytosed into oocytes within yolk granules during the vitellogenic stages of oogenesis. Mutations that disrupt yolk uptake by oocytes inhibit vertical Spiroplasma transmission and lead to an accumulation of these bacteria outside the oocyte. Impairment of yolk secretion by the fat body results in Spiroplasma not reaching the oocyte and a severe reduction of vertical transmission. We propose a model in which Spiroplasma first interacts with yolk in the hemolymph to gain access to the oocyte and then uses the yolk receptor, Yolkless, to be endocytosed into the oocyte. Cooption of the yolk uptake machinery is a powerful strategy for endosymbionts to target the germ line and achieve vertical transmission. This mechanism may apply to other endosymbionts and provides a possible explanation for endosymbiont host specificity. IMPORTANCE: Most insect species, including important disease vectors and crop pests, harbor vertically transmitted endosymbiotic bacteria. Studies have shown that many facultative endosymbionts, including Spiroplasma, confer protection against different classes of parasites on their hosts and therefore are attractive tools for the control of vector-borne diseases. The ability to be efficiently transmitted from females to their offspring is the key feature shaping associations between insects and their inherited endosymbionts, but to date, little is known about the mechanisms involved. In oviparous animals, yolk accumulates in developing eggs and serves to meet the nutritional demands of embryonic development. Here we show that Spiroplasma coopts the yolk transport and uptake machinery to colonize the germ line and ensure efficient vertical transmission. The uptake of yolk is a female germ line-specific feature and therefore an attractive target for cooption by endosymbionts that need to maintain high-fidelity maternal transmission

The Role of Lipid Competition for Endosymbiont-Mediated Protection against Parasitoid Wasps in Drosophila

Insects commonly harbor facultative bacterial endosymbionts, such as Wolbachia and Spiroplasma species, that are vertically transmitted from mothers to their offspring. These endosymbiontic bacteria increase their propagation by manipulating host reproduction or by protecting their hosts against natural enemies. While an increasing number of studies have reported endosymbiont-mediated protection, little is known about the mechanisms underlying this protection. Here, we analyze the mechanisms underlying protection from parasitoid wasps in Drosophila melanogaster mediated by its facultative endosymbiont Spiroplasma poulsonii. Our results indicate that S. poulsonii exerts protection against two distantly related wasp species, Leptopilina boulardi and Asobara tabida. S. poulsonii-mediated protection against parasitoid wasps takes place at the pupal stage and is not associated with an increased cellular immune response. In this work, we provide three important observations that support the notion that S. poulsonii bacteria and wasp larvae compete for host lipids and that this competition underlies symbiont-mediated protection. First, lipid quantification shows that both S. poulsonii and parasitoid wasps deplete D. melanogaster hemolymph lipids. Second, the depletion of hemolymphatic lipids using the Lpp RNA interference (Lpp RNAi) construct reduces wasp success in larvae that are not infected with S. poulsonii and blocks S. poulsonii growth. Third, we show that the growth of S. poulsonii bacteria is not affected by the presence of the wasps, indicating that when S. poulsonii is present, larval wasps will develop in a lipid-depleted environment. We propose that competition for host lipids may be relevant to endosymbiont-mediated protection in other systems and could explain the broad spectrum of protection provide

Differential immune responses in new and old fruit fly-parasitoid associations : implications for their management

The oriental fruit fly, Bactrocera dorsalis (Hendel), and marula fruit fly, Ceratitis cosyra (Walker), are major fruit-infesting tephritids across sub-Saharan Africa. Biological control of these pests using parasitic wasps has been widely adopted but with varying levels of success. Most studies investigating host-parasitoid models have focused on functional and evolutionary aspects leaving a knowledge gap about the physiological mechanisms underpinning the efficacy of parasitoids as biocontrol agents of tephritids. To better understand these physiological mechanisms, we investigated changes in the cellular immune responses of C. cosyra and B. dorsalis when exposed to the parasitic wasps, Diachasmimorpha longicaudata (Ashmaed) and Psyttalia cosyrae (Wilkinson). We found that B. dorsalis was more resistant to parasitisation, had a higher hemocyte count, and encapsulated more parasitoid eggs compared to C. cosyra, achieving up to 100% encapsulation when exposed to P. cosyrae. Exposing B. dorsalis to either parasitoid species induced the formation of a rare cell type, the giant multinucleated hemocyte, which was not observed in C. cosyra. Furthermore, compared to P. cosyrae-parasitized larvae, those of both host species parasitized by D. longicaudata had lower encapsulation rates, hemocyte counts and spreading abilities and yielded a higher number of parasitoid progeny with the highest parasitoid emergence (72.13%) recorded in C. cosyra. These results demonstrate that cellular immune responses are central to host-parasitoid interaction in tephritid fruit flies and further suggest that D. longicaudata presents greater potential as a biocontrol agent of B. dorsalis and C. cosyra in horticultural cropping systems.DATA AVAILABILITY STATEMENT: The datasets presented in this study can be found in online repositories. The names of the repository/repositories and accession number(s) can be found below: https://dmmg.icipe. org/dataportal/dataset/differential-immune-responses-in-newand-old-fruit-fly-parasitoid-association.International Development Research Centre (IDRC) and the Australian Centre for International Agricultural Research (ACIAR) for the project “Alien invasive fruit flies in Southern Africa: Implementation of a sustainable IPM programme to combat".https://www.frontiersin.org/journals/physiologydm2022Zoology and Entomolog

The Malaria Cell Atlas: single parasite transcriptomes across the complete Plasmodium life cycle

Malaria parasites adopt a remarkable variety of morphological life stages as they transition through multiple mammalian host and mosquito vector environments. We profiled the single-cell transcriptomes of thousands of individual parasites, deriving the first high-resolution transcriptional atlas of the entire life cycle. We then used our atlas to precisely define developmental stages of single cells from three different human malaria parasite species, including parasites isolated directly from infected individuals. The Malaria Cell Atlas provides both a comprehensive view of gene usage in a eukaryotic parasite and an open-access reference dataset for the study of malaria parasites

Spiroplasma and host immunity: activation of humoral immune responses increases endosymbiont load and susceptibility to certain Gram‐negative bacterial pathogens in Drosophila melanogaster

Spiroplasma poulsonii and its relatives are facultative, vertically transmitted endosymbionts harboured by several Drosophila species. Their long‐term survival requires not only evasion of host immunity, but also that Spiroplasma does not have a net detrimental effect on host fitness. These requirements provide the central framework for interactions between host and endosymbiont. We use Drosophila melaogaster as a model to unravel aspects of the mechanistic basis of endosymbiont–host immune interactions. Here we show that Spiroplasma does not activate an immune response in Drosophila and is not susceptible to either the cellular or humoral arms of the Drosophila immune system. We gain unexpected insight into host factors that can promote Spiroplasma growth by showing that activation of Toll and Imd immune pathways actually increases Sprioplasma titre. Spiroplasma‐mediated protection is not observed for variety of fungal and bacterial pathogens and Spiroplasma actually increases susceptibility of Drosophila to certain Gram‐negative pathogens. Finally, we show that the growth of endosymbiotic Spiroplasma is apparently self‐regulated, as suggested by the unhindered proliferation of non‐endosymbiotic Spiroplasma citri in fly haemolymph

Population genetics of sexually antagonistic mitochondrial mutants under inbreeding

In random mating populations, the fate of mitochondrial mutations with sexually antagonistic effects in males and females is based solely on their effects in females. Therefore, mitochondrial mutations that are beneficial for females but deleterious for males will be fixed in a deterministic model. Why then are males not less fertile? One among many several explanations is that inbreeding limits the ability of mutants to spread since the fitness of a mother is now linked to her son's fertility. We model this situation analytically and determine conditions under which such sexually antagonistic mitochondrial mutants can spread and fix in a population. We also provide alternative hypotheses for the lack of observed male sterility in natural populations

Wolbachia as populations within individual insects: causes and consequences of density variation in natural populations

The population-level dynamics of maternally transmitted endosymbionts, including reproductive parasites, depends primarily on the fitness effects and transmission fidelity of these infections. Although experimental laboratory studies have shown that within-host endosymbiont density can affect both of these factors, the existence of such effects in natural populations has not yet been documented. Using quantitative PCR, we survey the density of male-killing Wolbachia in natural populations of Drosophila innubila females from the Chiricahua Mountains of Arizona. We find that there is substantial (20 000-fold) variation in Wolbachia density among wild flies and that within-host Wolbachia density is positively correlated with both the efficacy of male killing and maternal transmission fidelity. Mean Wolbachia density increases three- to five-fold from early to late in the season. This pattern suggests that Wolbachia density declines with fly age, a conclusion corroborated by a laboratory study of Wolbachia density as a function of age. Finally, we suggest three alternative hypotheses to account for the approximately lognormal distribution of Wolbachia density among wild flies

Parasitic elements, extinction and adaptation : theoretical and empirical investigations

Thesis (Ph. D.)--University of Rochester. Dept. of Biology, 2011. Chapters 1-2 were co-authored with H. Allen Orr. Chapter 3 was co-authored with John Jaenike, Jeremy K. Herren, and Lisa M. Boelio. Chapter 4 was co-authored with John Jaenike. John Jaenike assisted with writing chapter 5.My dissertation research uses several different approaches to investigate evolutionary and evolutionary ecology questions. In Chapter 1, I investigate the likelihood that a population can save itself from extinction via spread of a rare beneficial allele after an environmental change. Although populations can, in principle, save themselves using new beneficial mutations or beneficial mutations from the standing genetic variation, I found, using a population genetic approach, that it is much more likely that they use standing genetic variation. This is particularly true if there are several copies of the beneficial mutation in the standing variation, making it unlikely that all are lost due to chance. In Chapter 2, I derive analytical solutions to predict the trajectory of evolutionary change under natural selection. Given that a population is predominated by a suboptimal genotype and that there are several beneficial mutations available, the analytical solutions in chapter 2 give the probability that the population fixes a particular set of beneficial mutations and the probability that it does so in a particular order. Generally, evolutionary trajectories are more predictable when there is greater variation in selection coefficients of beneficial mutations. My attention turns to empirical work on Drosophila innubila in chapters 3 through 5. About 1/3 of female D. innubila are infected with maternally-transmitted male-killing Wolbachia. In Chapter 3 I find that there is considerable seasonal variation in Wolbachia density in natural populations and that density correlates with both penetrance of male-killing and efficiency of transmission in the wild. Several alternatives for the distribution of densities among flies are discussed. In Chapter 4 I investigate what factors help maintain the Wolbachia infection. To persist, the Wolbachia must either supplement vertical transmission with horizontal transmission or benefit its host through the act of killing males or in some way unrelated to male-killing. I found that neither horizontal transmission nor male-killing-dependent benefits could explain the prevalence of infection. Instead, a combination of male-killing-dependent and -independent factors, including virus protection and nutritional provisioning, likely maintain the infection. Since Wolbachia in D. innubila protect against RNA viruses, as shown in Chapter 4, in Chapter 5 I attempted to determine what viruses are important in natural populations. I begin with a metagenomic survey to discover viruses. The most common novel virus was a DNA virus related to the Nudiviruses. This virus is phylogenetically widespread and is associated with significant fitness costs to its host