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Sample records for influential passenger wolbachia

  1. Species in Wolbachia? Proposal for the designation of 'Candidatus Wolbachia bourtzisii', 'Candidatus Wolbachia onchocercicola', 'Candidatus Wolbachia blaxteri', 'Candidatus Wolbachia brugii', 'Candidatus Wolbachia taylori', 'Candidatus Wolbachia collembolicola' and 'Candidatus Wolbachia multihospitum' for the different species within Wolbachia supergroups.

    PubMed

    Ramírez-Puebla, Shamayim T; Servín-Garcidueñas, Luis E; Ormeño-Orrillo, Ernesto; Vera-Ponce de León, Arturo; Rosenblueth, Mónica; Delaye, Luis; Martínez, Julio; Martínez-Romero, Esperanza

    2015-09-01

    Wolbachia are highly extended bacterial endosymbionts that infect arthropods and filarial nematodes and produce contrasting phenotypes on their hosts. Wolbachia taxonomy has been understudied. Currently, Wolbachia strains are classified into phylogenetic supergroups. Here we applied phylogenomic analyses to study Wolbachia evolutionary relationships and examined metrics derived from their genome sequences such as average nucleotide identity (ANI), in silico DNA-DNA hybridization (DDH), G+C content, and synteny to shed light on the taxonomy of these bacteria. Draft genome sequences of strains wDacA and wDacB obtained from the carmine cochineal insect Dactylopius coccus were included. Although all analyses indicated that each Wolbachia supergroup represents a distinct evolutionary lineage, we found that some of the analyzed supergroups showed enough internal heterogeneity to be considered as assemblages of more than one species. Thus, supergroups would represent supraspecific groupings. Consequently, Wolbachia pipientis nomen species would apply only to strains of supergroup B and we propose the designation of 'Candidatus Wolbachia bourtzisii', 'Candidatus Wolbachia onchocercicola', 'Candidatus Wolbachia blaxterii', 'Candidatus Wolbachia brugii', 'Candidatus Wolbachia taylorii', 'Candidatus Wolbachia collembolicola' and 'Candidatus Wolbachia multihospitis' for other supergroups.

  2. "Disproportionately Influential?"

    ERIC Educational Resources Information Center

    Stuart, Reginald

    2010-01-01

    This article discusses why the Lumina Foundation is considered so influential in higher education despite its small size and the fact that is is a relatively new foundation. Lumina approaches its 10th anniversary this month with a focused higher education funding mission targeting efforts aimed at expanding access and success beyond high school,…

  3. Wolbachia versus dengue

    PubMed Central

    Bull, James J.; Turelli, Michael

    2013-01-01

    A novel form of biological control is being applied to the dengue virus. The agent is the maternally transmitted bacterium Wolbachia, naturally absent from the main dengue vector, the mosquito Aedes aegypti. Three Wolbachia-based control strategies have been proposed. One is suppression of mosquito populations by large-scale releases of males incompatible with native females; this intervention requires ongoing releases. The other interventions transform wild mosquito populations with Wolbachia that spread via the frequency-dependent fitness advantage of Wolbachia-infected females; those interventions potentially require just a single, local release for area-wide disease control. One of these latter strategies uses Wolbachia that shortens mosquito life, indirectly preventing viral maturation/transmission. The other strategy uses Wolbachia that block viral transmission. All interventions can be undermined by viral, bacterial or mosquito evolution; viral virulence in humans may also evolve. We examine existing theory, experiments and comparative evidence to motivate predictions about evolutionary outcomes. (i) The life-shortening strategy seems the most likely to be thwarted by evolution. (ii) Mosquito suppression has a reasonable chance of working locally, at least in the short term, but long-term success over large areas is challenging. (iii) Dengue blocking faces strong selection for viral resistance but may well persist indefinitely at some level. Virulence evolution is not mathematically predictable, but comparative data provide no precedent for Wolbachia increasing dengue virulence. On balance, our analysis suggests that the considerable possible benefits of these technologies outweigh the known negatives, but the actual risk is largely unknown. PMID:24481199

  4. Wolbachia modulates Chikungunya replication in Aedes albopictus.

    PubMed

    Mousson, L; Martin, E; Zouache, K; Madec, Y; Mavingui, P; Failloux, A B

    2010-05-01

    The Aedes albopictus mosquito has been involved as the principal vector of recent major outbreaks due to the chikungunya virus (CHIKV). The species is naturally infected by two strains of Wolbachia (wAlbA and wAlbB). Wolbachia infections are thought to have spread by manipulating the reproduction of their hosts; cytoplasmic incompatibility is the mechanism used by Wolbachia to invade natural populations of many insects including Ae. albopictus. Here, we report a study on the effects of removing Wolbachia from Ae. albopictus on CHIKV replication and examine the consequences of CHIKV infection on some life-history traits (survival and reproduction) of Wolbachia-free Ae. albopictus. We found that Wolbachia-free mosquitoes maintained a highly heterogeneous CHIKV replication compared to Wolbachia-infected individuals. In Wolbachia-infected Ae. albopictus, the regular increase of CHIKV followed by a steady viral load from day 4 post-infection onwards was concomitant with a decline in Wolbachia density. This profile was also detected when examining the two key organs for viral transmission, the midgut and the salivary glands. Moreover, Wolbachia-free Ae. albopictus was not altered in life-history traits such as survival, oviposition and hatching characteristics whether infected or not with CHIKV. We found that Wolbachia is not essential for viral replication, its presence could lead to optimize replication from day 4 post-infection onwards, coinciding with a decrease in Wolbachia density. Wolbachia may regulate viral replication in Ae. albopictus, with consequences on survival and reproduction.

  5. The rich somatic life of Wolbachia.

    PubMed

    Pietri, Jose E; DeBruhl, Heather; Sullivan, William

    2016-12-01

    Wolbachia is an intracellular endosymbiont infecting most arthropod and some filarial nematode species that is vertically transmitted through the maternal lineage. Due to this primary mechanism of transmission, most studies have focused on Wolbachia interactions with the host germline. However, over the last decade many studies have emerged highlighting the prominence of Wolbachia in somatic tissues, implicating somatic tissue tropism as an important aspect of the life history of this endosymbiont. Here, we review our current understanding of Wolbachia-host interactions at both the cellular and organismal level, with a focus on Wolbachia in somatic tissues.

  6. Wolbachia in Neotropical terrestrial isopods.

    PubMed

    Zimmermann, Bianca L; Bouchon, Didier; Almerão, Maurício P; Araujo, Paula B

    2015-04-01

    Despite Wolbachia being widespread among terrestrial isopods, studies on this symbiotic relationship are still incipient in the Neotropical region. The aims of the present study were to investigate the presence and prevalence of Wolbachia in natural populations of terrestrial isopod species in South America, and to analyze the diversity and phylogenetic relationships of Wolbachia strains. A total of 1172 individuals representing 11 families and 35 species were analyzed. We observed distinct evolutionary scenarios according to the geographical origins of the species: strains harbored by most of the introduced species belong to the Oniclade in supergroup B and are identical to those found in their original ecozone (i.e. Palearctic). On the other hand, the strains found in native Neotropical terrestrial isopods showed low prevalence, high diversity and none of them belonged to the Oniclade, although most belonged to supergroup B. The dynamics of infection in Neotropical species seems to be the result of several events of loss and acquisition of the bacteria, which refutes the hypothesis of an ancestral acquisition of Wolbachia in Oniscidea. The presence of strains from supergroups A and F was also detected for the first time in terrestrial isopods, revealing a Wolbachia diversity previously unknown for this group of host.

  7. Uncovering Wolbachia diversity upon artificial host transfer.

    PubMed

    Schneider, Daniela I; Riegler, Markus; Arthofer, Wolfgang; Merçot, Hervé; Stauffer, Christian; Miller, Wolfgang J

    2013-01-01

    The common endosymbiotic Wolbachia bacteria influence arthropod hosts in multiple ways. They are mostly recognized for their manipulations of host reproduction, yet, more recent studies demonstrate that Wolbachia also impact host behavior, metabolic pathways and immunity. Besides their biological and evolutionary roles, Wolbachia are new potential biological control agents for pest and vector management. Importantly, Wolbachia-based control strategies require controlled symbiont transfer between host species and predictable outcomes of novel Wolbachia-host associations. Theoretically, this artificial horizontal transfer could inflict genetic changes within transferred Wolbachia populations. This could be facilitated through de novo mutations in the novel recipient host or changes of haplotype frequencies of polymorphic Wolbachia populations when transferred from donor to recipient hosts. Here we show that Wolbachia resident in the European cherry fruit fly, Rhagoletis cerasi, exhibit ancestral and cryptic sequence polymorphism in three symbiont genes, which are exposed upon microinjection into the new hosts Drosophila simulans and Ceratitis capitata. Our analyses of Wolbachia in microinjected D. simulans over 150 generations after microinjection uncovered infections with multiple Wolbachia strains in trans-infected lines that had previously been typed as single infections. This confirms the persistence of low-titer Wolbachia strains in microinjection experiments that had previously escaped standard detection techniques. Our study demonstrates that infections by multiple Wolbachia strains can shift in prevalence after artificial host transfer driven by either stochastic or selective processes. Trans-infection of Wolbachia can claim fitness costs in new hosts and we speculate that these costs may have driven the shifts of Wolbachia strains that we saw in our model system.

  8. Wolbachia Sequence Typing in Butterflies Using Pyrosequencing.

    PubMed

    Choi, Sungmi; Shin, Su-Kyoung; Jeong, Gilsang; Yi, Hana

    2015-09-01

    Wolbachia is an obligate symbiotic bacteria that is ubiquitous in arthropods, with 25-70% of insect species estimated to be infected. Wolbachia species can interact with their insect hosts in a mutualistic or parasitic manner. Sequence types (ST) of Wolbachia are determined by multilocus sequence typing (MLST) of housekeeping genes. However, there are some limitations to MLST with respect to the generation of clone libraries and the Sanger sequencing method when a host is infected with multiple STs of Wolbachia. To assess the feasibility of massive parallel sequencing, also known as next-generation sequencing, we used pyrosequencing for sequence typing of Wolbachia in butterflies. We collected three species of butterflies (Eurema hecabe, Eurema laeta, and Tongeia fischeri) common to Korea and screened them for Wolbachia STs. We found that T. fischeri was infected with a single ST of Wolbachia, ST41. In contrast, E. hecabe and E. laeta were each infected with two STs of Wolbachia, ST41 and ST40. Our results clearly demonstrate that pyrosequencing-based MLST has a higher sensitivity than cloning and Sanger sequencing methods for the detection of minor alleles. Considering the high prevalence of infection with multiple Wolbachia STs, next-generation sequencing with improved analysis would assist with scaling up approaches to Wolbachia MLST.

  9. Oxidative stress correlates with Wolbachia-mediated antiviral protection in Wolbachia-Drosophila associations.

    PubMed

    Wong, Zhee Sheen; Brownlie, Jeremy C; Johnson, Karyn N

    2015-05-01

    Wolbachia mediates antiviral protection in insect hosts and is being developed as a potential biocontrol agent to reduce the spread of insect-vectored viruses. Definition of the molecular mechanism that generates protection is important for understanding the tripartite interaction between host insect, Wolbachia, and virus. Elevated oxidative stress was previously reported for a mosquito line experimentally infected with Wolbachia, suggesting that oxidative stress is important for Wolbachia-mediated antiviral protection. However, Wolbachia experimentally introduced into mosquitoes impacts a range of host fitness traits, some of which are unrelated to antiviral protection. To explore whether elevated oxidative stress is associated with antiviral protection in Wolbachia-infected insects, we analyzed oxidative stress of five Wolbachia-infected Drosophila lines. In flies infected with protective Wolbachia strains, hydrogen peroxide concentrations were 1.25- to 2-fold higher than those in paired fly lines cured of Wolbachia infection. In contrast, there was no difference in the hydrogen peroxide concentrations in flies infected with nonprotective Wolbachia strains compared to flies cured of Wolbachia infection. Using a Drosophila mutant that produces increased levels of hydrogen peroxide, we investigated whether flies with high levels of endogenous reactive oxygen species had altered responses to virus infection and found that flies with high levels of endogenous hydrogen peroxide were less susceptible to virus-induced mortality. Taken together, these results suggest that elevated oxidative stress correlates with Wolbachia-mediated antiviral protection in natural Drosophila hosts.

  10. 77 FR 27054 - Wolbachia pipientis;

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-05-08

    ... pesticide of insects/mosquitoes. The Agency has determined that the permit may be of regional and national... release of male Aedes polynesienis mosquitoes infected with Wolbachia pipientis in American Samoa. The male mosquitoes will mate with indigenous female Aedes polynesienis, causing conditional sterility...

  11. Wolbachia spread dynamics in stochastic environments.

    PubMed

    Hu, Linchao; Huang, Mugen; Tang, Moxun; Yu, Jianshe; Zheng, Bo

    2015-12-01

    Dengue fever is a mosquito-borne viral disease with 100 million people infected annually. A novel strategy for dengue control uses the bacterium Wolbachia to invade dengue vector Aedes mosquitoes. As the impact of environmental heterogeneity on Wolbachia spread dynamics in natural areas has been rarely quantified, we develop a model of differential equations for which the environmental conditions switch randomly between two regimes. We find some striking phenomena that random regime transitions could drive Wolbachia to extinction from certain initial states confirmed Wolbachia fixation in homogeneous environments, and mosquito releasing facilitates Wolbachia invasion more effectively when the regimes transit frequently. By superimposing the phase spaces of the ODE systems defined in each regime, we identify the threshold curves below which Wolbachia invades the whole population, which extends the theory of threshold infection frequency to stochastic environments.

  12. Supergroup F Wolbachia bacteria parasitise lice (Insecta: Phthiraptera).

    PubMed

    Covacin, Catherine; Barker, Stephen C

    2007-02-01

    We studied six species of lice from three of the four suborders of lice. These lice were infected with Wolbachia bacteria from supergroups A and F. This is the first report of an infection of supergroup F Wolbachia in lice. To date, Wolbachia from supergroup F have been found in filarial nematodes, Mansonella spp., and, rarely, in insects. We inferred the phylogeny of the Wolbachia from lice and representatives of all Wolbachia supergroups, with nucleotide sequences from the small subunit ribosomal RNA gene (SSU rRNA). There was no evidence of congruence between the taxon of louse and the Wolbachia bacteria that infect lice. There is no evidence that Wolbachia and their louse hosts co-evolved at least at the level of Wolbachia supergroups. We propose a novel mechanism for the horizontal transfer of Wolbachia between different species of lice from birds: transfer of Wolbachia during phoresis by hippoboscid flies.

  13. Wolbachia infection and Lepidoptera of conservation concern.

    PubMed

    Hamm, C A; Handley, C A; Pike, A; Forister, M L; Fordyce, J A; Nice, C C

    2014-01-14

    Conservation of at-risk species requires multi-faceted and carefully-considered management approaches to be successful. For arthropods, the presence of endosymbiotic bacteria, such as Wolbachia (Rickettsiales: Rickettsiaceae), may complicate management plans and exacerbate the challenges faced by conservation managers. Wolbachia poses a substantial and underappreciated threat to the conservation of arthropods because infection may induce a number of phenotypic effects, most of which are considered deleterious to the host population. In this study, the prevalence of Wolbachia infection in lepidopteran species of conservation concern was examined. Using standard molecular techniques, 22 species of Lepidoptera were screened, of which 19 were infected with Wolbachia. This rate is comparable to that observed in insects as a whole. However, this is likely an underestimate because geographic sampling was not extensive and may not have included infected segments of the species' ranges. Wolbachia infections may be particularly problematic for conservation management plans that incorporate captive propagation or translocation. Inadvertent introduction of Wolbachia into uninfected populations or introduction of a new strain may put these populations at greater risk for extinction. Further sampling to investigate the geographic extent of Wolbachia infections within species of conservation concern and experiments designed to determine the nature of the infection phenotype(s) are necessary to manage the potential threat of infection.

  14. Wolbachia endosymbionts and human disease control.

    PubMed

    Slatko, Barton E; Luck, Ashley N; Dobson, Stephen L; Foster, Jeremy M

    2014-07-01

    Most human filarial nematode parasites and arthropods are hosts for a bacterial endosymbiont, Wolbachia. In filaria, Wolbachia are required for normal development, fertility and survival, whereas in arthropods, they are largely parasitic and can influence development and reproduction, but are generally not required for host survival. Due to their obligate nature in filarial parasites, Wolbachia have been a target for drug discovery initiatives using several approaches including diversity and focused library screening and genomic sequence analysis. In vitro and in vivo anti-Wolbachia antibiotic treatments have been shown to have adulticidal activity, a long sought goal of filarial parasite drug discovery. In mosquitoes, it has been shown that the presence of Wolbachia can inhibit the transmission of certain viruses, such as Dengue, Chikungunya, Yellow Fever, West Nile, as well as the infectivity of the malaria-causing protozoan, Plasmodium and filarial nematodes. Furthermore, Wolbachia can cause a form of conditional sterility that can be used to suppress populations of mosquitoes and additional medically important insects. Thus Wolbachia, a pandemic endosymbiont offers great potential for elimination of a wide-variety of devastating human diseases.

  15. Iron Necessity: The Secret of Wolbachia's Success?

    PubMed Central

    Gill, Alessandra Christina; Darby, Alistair C.; Makepeace, Benjamin L.

    2014-01-01

    The bacterium Wolbachia (order Rickettsiales) is probably the world's most successful vertically-transmitted symbiont, distributed among a staggering 40% of terrestrial arthropod species. Wolbachia has great potential in vector control due to its ability to manipulate its hosts' reproduction and to impede the replication and dissemination of arboviruses and other pathogens within haematophagous arthropods. In addition, the unexpected presence of Wolbachia in filarial nematodes of medical and veterinary importance has provided an opportunity to target the adult worms of Wuchereria bancrofti, Onchocerca volvulus, and Dirofilaria immitis with safe drugs such as doxycycline. A striking feature of Wolbachia is its phenotypic plasticity between (and sometimes within) hosts, which may be underpinned by its ability to integrate itself into several key processes within eukaryotic cells: oxidative stress, autophagy, and apoptosis. Importantly, despite significant differences in the genomes of arthropod and filarial Wolbachia strains, these nexuses appear to lie on a continuum in different hosts. Here, we consider how iron metabolism may represent a fundamental aspect of host homeostasis that is impacted by Wolbachia infection, connecting disparate pathways ranging from the provision of haem and ATP to programmed cell death, aging, and the recycling of intracellular resources. Depending on how Wolbachia and host cells interact across networks that depend on iron, the gradient between parasitism and mutualism may shift dynamically in some systems, or alternatively, stabilise on one or the other end of the spectrum. PMID:25329055

  16. Parthenogenesis induction by Wolbachia in parasitoids

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The maternally inherited intracellular bacterium, Wolbachia pipientis, induces reproductive alterations in host arthropod populations such as male-killing, feminization, parthenogenesis induction (PI, or reproduction without males) and cytoplasmic incompatibility. Here we report that PI in Mexican O...

  17. Mutualism Breakdown by Amplification of Wolbachia Genes

    PubMed Central

    Chrostek, Ewa; Teixeira, Luis

    2015-01-01

    Most insect species are associated with vertically transmitted endosymbionts. Because of the mode of transmission, the fitness of these symbionts is dependent on the fitness of the hosts. Therefore, these endosymbionts need to control their proliferation in order to minimize their cost for the host. The genetic bases and mechanisms of this regulation remain largely undetermined. The maternally inherited bacteria of the genus Wolbachia are the most common endosymbionts of insects, providing some of them with fitness benefits. In Drosophila melanogaster, Wolbachia wMelPop is a unique virulent variant that proliferates massively in the hosts and shortens their lifespan. The genetic bases of wMelPop virulence are unknown, and their identification would allow a better understanding of how Wolbachia levels are regulated. Here we show that amplification of a region containing eight Wolbachia genes, called Octomom, is responsible for wMelPop virulence. Using Drosophila lines selected for carrying Wolbachia with different Octomom copy numbers, we demonstrate that the number of Octomom copies determines Wolbachia titers and the strength of the lethal phenotype. Octomom amplification is unstable, and reversion of copy number to one reverts all the phenotypes. Our results provide a link between genotype and phenotype in Wolbachia and identify a genomic region regulating Wolbachia proliferation. We also prove that these bacteria can evolve rapidly. Rapid evolution by changes in gene copy number may be common in endosymbionts with a high number of mobile elements and other repeated regions. Understanding wMelPop pathogenicity and variability also allows researchers to better control and predict the outcome of releasing mosquitoes transinfected with this variant to block human vector-borne diseases. Our results show that transition from a mutualist to a pathogen may occur because of a single genomic change in the endosymbiont. This implies that there must be constant selection on

  18. Mutualism breakdown by amplification of Wolbachia genes.

    PubMed

    Chrostek, Ewa; Teixeira, Luis

    2015-02-01

    Most insect species are associated with vertically transmitted endosymbionts. Because of the mode of transmission, the fitness of these symbionts is dependent on the fitness of the hosts. Therefore, these endosymbionts need to control their proliferation in order to minimize their cost for the host. The genetic bases and mechanisms of this regulation remain largely undetermined. The maternally inherited bacteria of the genus Wolbachia are the most common endosymbionts of insects, providing some of them with fitness benefits. In Drosophila melanogaster, Wolbachia wMelPop is a unique virulent variant that proliferates massively in the hosts and shortens their lifespan. The genetic bases of wMelPop virulence are unknown, and their identification would allow a better understanding of how Wolbachia levels are regulated. Here we show that amplification of a region containing eight Wolbachia genes, called Octomom, is responsible for wMelPop virulence. Using Drosophila lines selected for carrying Wolbachia with different Octomom copy numbers, we demonstrate that the number of Octomom copies determines Wolbachia titers and the strength of the lethal phenotype. Octomom amplification is unstable, and reversion of copy number to one reverts all the phenotypes. Our results provide a link between genotype and phenotype in Wolbachia and identify a genomic region regulating Wolbachia proliferation. We also prove that these bacteria can evolve rapidly. Rapid evolution by changes in gene copy number may be common in endosymbionts with a high number of mobile elements and other repeated regions. Understanding wMelPop pathogenicity and variability also allows researchers to better control and predict the outcome of releasing mosquitoes transinfected with this variant to block human vector-borne diseases. Our results show that transition from a mutualist to a pathogen may occur because of a single genomic change in the endosymbiont. This implies that there must be constant selection on

  19. Phylogenomic analyses uncover origin and spread of the Wolbachia pandemic.

    PubMed

    Gerth, Michael; Gansauge, Marie-Theres; Weigert, Anne; Bleidorn, Christoph

    2014-10-06

    Of all obligate intracellular bacteria, Wolbachia is probably the most common. In general, Wolbachia are either widespread, opportunistic reproductive parasites of arthropods or essential mutualists in a single group of filarial nematodes, including many species of medical significance. To date, a robust phylogenetic backbone of Wolbachia is lacking and consequently, many Wolbachia-related phenomena cannot be discussed in a broader evolutionary context. Here we present the first comprehensive phylogenomic analysis of Wolbachia supergroup relationships based on new whole-genome-shotgun data. Our results suggest that Wolbachia has switched between its two major host groups at least twice. The ability of some arthropod-infecting Wolbachia to universally infect and to adapt to a broad range of hosts quickly is restricted to a single monophyletic lineage (containing supergroups A and B). Thus, the currently observable pandemic has likely a single evolutionary origin and is unique within the radiation of Wolbachia strains.

  20. Wolbachia-infection differs among potato psyllid haplotypes

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Wolbachia is a bacterial endosymbiont of insects that can manipulate insect reproduction. In many insects, Wolbachia-free females cannot produce viable offspring when mated by infected males. The manipulation of insect reproduction by Wolbachia has important implications for insect evolution and pop...

  1. Evidence for horizontal transfer of Wolbachia by a Drosophila mite.

    PubMed

    Brown, Amy N; Lloyd, Vett K

    2015-07-01

    Mites are common ectoparasites of Drosophila and have been implicated in bacterial and mobile element invasion of Drosophila stocks. The obligate endobacterium, Wolbachia, has widespread effects on gene expression in their arthropod hosts and alters host reproduction to enhance its survival and propagation, often with deleterious effects in Drosophila hosts. To determine whether Wolbachia could be transferred between Drosophila melanogaster laboratory stocks by the mite Tyrophagus putrescentiae, mites were introduced to Wolbachia-infected Drosophila vials. These vials were kept adjacent to mite-free and Wolbachia-uninfected Drosophila stock vials. The Wolbachia infection statuses of the infected and uninfected flies were checked from generation 1 to 5. Results indicate that Wolbachia DNA could be amplified from mites infesting Wolbachia-infected fly stocks and infection in the previously uninfected stocks arose within generation 1 or 2, concomitant with invasion of mites from the Wolbachia-infected stock. A possible mechanism for the transfer of Wolbachia from flies to mites and vice versa, can be inferred from time-lapse photography of fly and mite interactions. We demonstrated that mites ingest Drosophila corpses, including Wolbachia-infected corpses, and Drosophila larva ingest mites, providing possible sources of Wolbachia infection and transfer. This research demonstrated that T. putrescentiae white mites can facilitate Wolbachia transfer between Drosophila stocks and that this may occur by ingestion of infected corpses. Mite-vectored Wolbachia transfer allows for rapid establishment of Wolbachia infection within a new population. This mode of Wolbachia introduction may be relevant in nature as well as in the laboratory, and could have a variety of biological consequences.

  2. A Wolbachia deubiquitylating enzyme induces cytoplasmic incompatibility.

    PubMed

    Beckmann, John F; Ronau, Judith A; Hochstrasser, Mark

    2017-03-01

    Wolbachia are obligate intracellular bacteria(1) that infect arthropods, including approximately two-thirds of insect species(2). Wolbachia manipulate insect reproduction by enhancing their inheritance through the female germline. The most common alteration is cytoplasmic incompatibility (CI)(3-5), where eggs from uninfected females fail to develop when fertilized by sperm from Wolbachia-infected males. By contrast, if female and male partners are both infected, embryos are viable. CI is a gene-drive mechanism impacting population structure(6) and causing reproductive isolation(7), but its molecular mechanism has remained unknown. We show that a Wolbachia deubiquitylating enzyme (DUB) induces CI. The CI-inducing DUB, CidB, cleaves ubiquitin from substrates and is encoded in a two-gene operon, and the other protein, CidA, binds CidB. Binding is strongest between cognate partners in cidA-cidB homologues. In transgenic Drosophila, the cidA-cidB operon mimics CI when sperm introduce it into eggs, and a catalytically inactive DUB does not induce sterility. Toxicity is recapitulated in yeast by CidB alone; this requires DUB activity but is rescued by coexpressed CidA. A paralogous operon involves a putative nuclease (CinB) rather than a DUB. Analogous binding, toxicity and rescue in yeast were observed. These results identify a CI mechanism involving interacting proteins that are secreted into germline cells by Wolbachia, and suggest new methods for insect control.

  3. Wolbachia increases susceptibility to Plasmodium infection in a natural system.

    PubMed

    Zélé, F; Nicot, A; Berthomieu, A; Weill, M; Duron, O; Rivero, A

    2014-03-22

    Current views about the impact of Wolbachia on Plasmodium infections are almost entirely based on data regarding artificially transfected mosquitoes. This work has shown that Wolbachia reduces the intensity of Plasmodium infections in mosquitoes, raising the exciting possibility of using Wolbachia to control or limit the spread of malaria. Whether natural Wolbachia infections have the same parasite-inhibiting properties is not yet clear. Wolbachia-mosquito combinations with a long evolutionary history are, however, key for understanding what may happen with Wolbachia-transfected mosquitoes after several generations of coevolution. We investigate this issue using an entirely natural mosquito-Wolbachia-Plasmodium combination. In contrast to most previous studies, which have been centred on the quantification of the midgut stages of Plasmodium, we obtain a measurement of parasitaemia that relates directly to transmission by following infections to the salivary gland stages. We show that Wolbachia increases the susceptibility of Culex pipiens mosquitoes to Plasmodium relictum, significantly increasing the prevalence of salivary gland stage infections. This effect is independent of the density of Wolbachia in the mosquito. These results suggest that naturally Wolbachia-infected mosquitoes may, in fact, be better vectors of malaria than Wolbachia-free ones.

  4. Wolbachia infection frequencies in insects: evidence of a global equilibrium?

    PubMed Central

    Werren, J H; Windsor, D M

    2000-01-01

    Wolbachia are a group of cytoplasmically inherited bacteria that cause reproduction alterations in arthropods, including parthenogenesis, reproductive incompatibility, feminization of genetic males and male killing. Previous general surveys of insects in Panama and Britain found Wolbachia to be common, occurring in 16-22% of species. Here, using similar polymerase chain reaction methods, we report that 19.3% of a sample of temperate North American insects are infected with Wolbachia, a frequency strikingly similar to frequencies found in two other studies in widely separated locales. The results may indicate a widespread equilibrium of Wolbachia infection frequencies in insects whose maintenance remains to be explained. Alternatively, Wolbachia may be increasing in global insect communities. Within each of the three geographic regions surveyed, Hymenoptera are more frequently infected with A group Wolbachia and Lepidoptera more frequently infected with B group Wolbachia. PMID:10972121

  5. Wolbachia transmission dynamics in Formica wood ants

    PubMed Central

    2008-01-01

    Background The role of Wolbachia endosymbionts in shaping the mitochondrial diversity of their arthropod host depends on the effects they have on host reproduction and on the mode of transmission of the bacteria. We have compared the sequence diversity of wsp (Wolbachia surface protein gene) and the host mtDNA in a group of Formica ant species that have diverged approximately 0.5 million years ago (MYA). The aim was to study the relationship of Wolbachia and its ant hosts in terms of vertical and horizontal transmission of the bacteria. Results All studied ant species were doubly infected with two Wolbachia strains (wFex1 and wFex4) all over their geographical distribution area in Eurasia. The most common haplotypes of these strains were identical with strains previously described from a more distantly related Formica ant, with an estimated divergence time of 3.5 – 4 MYA. Some strain haplotypes were associated to the same or closely related mtDNA haplotypes as expected under vertical transmission. However, in several cases the wsp haplotypes coexisted with distant mtDNA haplotypes, a pattern which is more compatible with horizontal transmission of the bacteria. Conclusion Two lines of evidence suggest that the sharing of Wolbachia strains by all F. rufa species is rather due to horizontal than vertical transmission. First, the fact that endosymbiont strains identical to those of F. rufa ants have been found in another species that diverged 3.5–4 MYA strongly suggests that horizontal transfer can and does occur between Formica ants. Second, the frequent sharing of identical Wolbachia strains by distant mitochondrial lineages within the F. rufa group further shows that horizontal transmission has occurred repeatedly. Nevertheless, our dataset also provides some evidence for longer-term persistence of infection, indicating that Wolbachia infection within this host clade has been shaped by both horizontal and vertical transmission of symbionts. The fact that all

  6. Gut microbiota in Drosophila melanogaster interacts with Wolbachia but does not contribute to Wolbachia-mediated antiviral protection.

    PubMed

    Ye, Yixin H; Seleznev, Andrei; Flores, Heather A; Woolfit, Megan; McGraw, Elizabeth A

    2017-02-01

    Animals experience near constant infection with microorganisms. A significant proportion of these microbiota reside in the alimentary tract. There is a growing appreciation for the roles gut microbiota play in host biology. The gut microbiota of insects, for example, have been shown to help the host overcome pathogen infection either through direct competition or indirectly by stimulating host immunity. These defenses may also be supplemented by coinfecting maternally inherited microbes such as Wolbachia. The presence of Wolbachia in a host can delay and/or reduce death caused by RNA viruses. Whether the gut microbiota of the host interacts with Wolbachia, or vice versa, the precise role of Wolbachia in antiviral protection is not known. In this study, we used 16S rDNA sequencing to characterise changes in gut microbiota composition in Drosophila melanogaster associated with Wolbachia infection and antibiotic treatment. We subsequently tested whether changes in gut composition via antibiotic treatment altered Wolbachia-mediated antiviral properties. We found that both antibiotics and Wolbachia significantly reduced the biodiversity of the gut microbiota without changing the total microbial load. We also showed that changing the gut microbiota composition with antibiotic treatment enhanced Wolbachia density but did not confer greater antiviral protection against Drosophila C virus to the host. We concluded there are significant interactions between Wolbachia and gut microbiota, but changing gut microbiota composition is not likely to be a means through which Wolbachia conveys antiviral protection to its host.

  7. The Impact of Wolbachia on Virus Infection in Mosquitoes

    PubMed Central

    Johnson, Karyn N.

    2015-01-01

    Mosquito-borne viruses such as dengue, West Nile and chikungunya viruses cause significant morbidity and mortality in human populations. Since current methods are not sufficient to control disease occurrence, novel methods to control transmission of arboviruses would be beneficial. Recent studies have shown that virus infection and transmission in insects can be impeded by co-infection with the bacterium Wolbachia pipientis. Wolbachia is a maternally inherited endosymbiont that is commonly found in insects, including a number of mosquito vector species. In Drosophila, Wolbachia mediates antiviral protection against a broad range of RNA viruses. This discovery pointed to a potential strategy to interfere with mosquito transmission of arboviruses by artificially infecting mosquitoes with Wolbachia. This review outlines research on the prevalence of Wolbachia in mosquito vector species and the impact of antiviral effects in both naturally and artificially Wolbachia-infected mosquitoes. PMID:26556361

  8. The Impact of Wolbachia on Virus Infection in Mosquitoes.

    PubMed

    Johnson, Karyn N

    2015-11-04

    Mosquito-borne viruses such as dengue, West Nile and chikungunya viruses cause significant morbidity and mortality in human populations. Since current methods are not sufficient to control disease occurrence, novel methods to control transmission of arboviruses would be beneficial. Recent studies have shown that virus infection and transmission in insects can be impeded by co-infection with the bacterium Wolbachia pipientis. Wolbachia is a maternally inherited endosymbiont that is commonly found in insects, including a number of mosquito vector species. In Drosophila, Wolbachia mediates antiviral protection against a broad range of RNA viruses. This discovery pointed to a potential strategy to interfere with mosquito transmission of arboviruses by artificially infecting mosquitoes with Wolbachia. This review outlines research on the prevalence of Wolbachia in mosquito vector species and the impact of antiviral effects in both naturally and artificially Wolbachia-infected mosquitoes.

  9. Wolbachia increases susceptibility to Plasmodium infection in a natural system

    PubMed Central

    Zélé, F.; Nicot, A.; Berthomieu, A.; Weill, M.; Duron, O.; Rivero, A.

    2014-01-01

    Current views about the impact of Wolbachia on Plasmodium infections are almost entirely based on data regarding artificially transfected mosquitoes. This work has shown that Wolbachia reduces the intensity of Plasmodium infections in mosquitoes, raising the exciting possibility of using Wolbachia to control or limit the spread of malaria. Whether natural Wolbachia infections have the same parasite-inhibiting properties is not yet clear. Wolbachia–mosquito combinations with a long evolutionary history are, however, key for understanding what may happen with Wolbachia-transfected mosquitoes after several generations of coevolution. We investigate this issue using an entirely natural mosquito–Wolbachia–Plasmodium combination. In contrast to most previous studies, which have been centred on the quantification of the midgut stages of Plasmodium, we obtain a measurement of parasitaemia that relates directly to transmission by following infections to the salivary gland stages. We show that Wolbachia increases the susceptibility of Culex pipiens mosquitoes to Plasmodium relictum, significantly increasing the prevalence of salivary gland stage infections. This effect is independent of the density of Wolbachia in the mosquito. These results suggest that naturally Wolbachia-infected mosquitoes may, in fact, be better vectors of malaria than Wolbachia-free ones. PMID:24500167

  10. Distribution and dynamics of Wolbachia infection in Malaysian Aedes albopictus.

    PubMed

    Joanne, Sylvia; Vythilingam, Indra; Yugavathy, Nava; Leong, Cherng-Shii; Wong, Meng-Li; AbuBakar, Sazaly

    2015-08-01

    Wolbachia are maternally transmitted bacteria found in most arthropods and nematodes, but little is known about their distribution and reproductive dynamics in the Malaysian dengue vector Aedes albopictus. In this study, polymerase chain reaction (PCR) was used to determine the presence of Wolbachia from field collected Ae. albopictus from various parts of the country using wsp specific primers. Ae. albopictus had Wolbachia infection ranging from 60 to 100%. No sequence diversity of wsp gene was found within all wAlbA and wAlbB sequences. Our findings suggest that Wolbachia infection amongst the Malaysian Ae. albopictus were not homogenously distributed in all districts in Malaysia. The presence of Wolbachia in different organs of Ae. albopictus was also determined. Wolbachia were only found in the ovaries and midguts of the mosquitoes, while absent in the salivary glands. The effects of Wolbachia on Ae. albopictus fecundity, longevity and egg viability were studied using infected and uninfected colonies. The removal of Wolbachia from Ae. albopictus resulted in reduced fecundity, longevity and egg viability, thus. Wolbachia seem to play a vital role in Ae. albopictus reproductive system.

  11. Wolbachia as a bacteriocyte-associated nutritional mutualist

    PubMed Central

    Hosokawa, Takahiro; Koga, Ryuichi; Kikuchi, Yoshitomo; Meng, Xian-Ying; Fukatsu, Takema

    2009-01-01

    Many insects are dependent on bacterial symbionts that provide essential nutrients (ex. aphid–Buchnera and tsetse–Wiglesworthia associations), wherein the symbionts are harbored in specific cells called bacteriocytes that constitute a symbiotic organ bacteriome. Facultative and parasitic bacterial symbionts like Wolbachia have been regarded as evolutionarily distinct from such obligate nutritional mutualists. However, we discovered that, in the bedbug Cimex lectularius, Wolbachia resides in a bacteriome and appears to be an obligate nutritional mutualist. Two bacterial symbionts, a Wolbachia strain and an unnamed γ-proteobacterium, were identified from different strains of the bedbug. The Wolbachia symbiont was detected from all of the insects examined whereas the γ-proteobacterium was found in a part of them. The Wolbachia symbiont was specifically localized in the bacteriomes and vertically transmitted via the somatic stem cell niche of germalia to oocytes, infecting the incipient symbiotic organ at an early stage of the embryogenesis. Elimination of the Wolbachia symbiont resulted in retarded growth and sterility of the host insect. These deficiencies were rescued by oral supplementation of B vitamins, confirming the essential nutritional role of the symbiont for the host. The estimated genome size of the Wolbachia symbiont was around 1.3 Mb, which was almost equivalent to the genome sizes of parasitic Wolbachia strains of other insects. These results indicate that bacteriocyte-associated nutritional mutualism can evolve from facultative and prevalent microbial associates like Wolbachia, highlighting a previously unknown aspect of the parasitism-mutualism evolutionary continuum. PMID:20080750

  12. Closely related Wolbachia recovered from different genera of Mexican Thelytokous figitidae (Hymenoptera)

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Closely related novel Wolbachia strains were recovered from the thelytokous figitids Odontosema anastrephae Borgmeier and Aganaspis alujai Ovruski et al. No Wolbachia were detected in a bi-sexual strain of O. anastrephae. While the presence or absence of Wolbachia does not demonstrate that Wolbachia...

  13. Wolbachia and DNA barcoding insects: patterns, potential, and problems.

    PubMed

    Smith, M Alex; Bertrand, Claudia; Crosby, Kate; Eveleigh, Eldon S; Fernandez-Triana, Jose; Fisher, Brian L; Gibbs, Jason; Hajibabaei, Mehrdad; Hallwachs, Winnie; Hind, Katharine; Hrcek, Jan; Huang, Da-Wei; Janda, Milan; Janzen, Daniel H; Li, Yanwei; Miller, Scott E; Packer, Laurence; Quicke, Donald; Ratnasingham, Sujeevan; Rodriguez, Josephine; Rougerie, Rodolphe; Shaw, Mark R; Sheffield, Cory; Stahlhut, Julie K; Steinke, Dirk; Whitfield, James; Wood, Monty; Zhou, Xin

    2012-01-01

    Wolbachia is a genus of bacterial endosymbionts that impacts the breeding systems of their hosts. Wolbachia can confuse the patterns of mitochondrial variation, including DNA barcodes, because it influences the pathways through which mitochondria are inherited. We examined the extent to which these endosymbionts are detected in routine DNA barcoding, assessed their impact upon the insect sequence divergence and identification accuracy, and considered the variation present in Wolbachia COI. Using both standard PCR assays (Wolbachia surface coding protein--wsp), and bacterial COI fragments we found evidence of Wolbachia in insect total genomic extracts created for DNA barcoding library construction. When >2 million insect COI trace files were examined on the Barcode of Life Datasystem (BOLD) Wolbachia COI was present in 0.16% of the cases. It is possible to generate Wolbachia COI using standard insect primers; however, that amplicon was never confused with the COI of the host. Wolbachia alleles recovered were predominantly Supergroup A and were broadly distributed geographically and phylogenetically. We conclude that the presence of the Wolbachia DNA in total genomic extracts made from insects is unlikely to compromise the accuracy of the DNA barcode library; in fact, the ability to query this DNA library (the database and the extracts) for endosymbionts is one of the ancillary benefits of such a large scale endeavor--which we provide several examples. It is our conclusion that regular assays for Wolbachia presence and type can, and should, be adopted by large scale insect barcoding initiatives. While COI is one of the five multi-locus sequence typing (MLST) genes used for categorizing Wolbachia, there is limited overlap with the eukaryotic DNA barcode region.

  14. Bacteria Endosymbiont, Wolbachia, Promotes Parasitism of Parasitoid Wasp Asobara japonica

    PubMed Central

    Matsumoto, Hitoshi; Hayakawa, Yoichi

    2015-01-01

    Wolbachia is the most widespread endosymbiotic bacterium that manipulates reproduction of its arthropod hosts to enhance its own spread throughout host populations. Infection with Wolbachia causes complete parthenogenetic reproduction in many Hymenoptera, producing only female offspring. The mechanism of such reproductive manipulation by Wolbachia has been extensively studied. However, the effects of Wolbachia symbiosis on behavioral traits of the hosts are scarcely investigated. The parasitoid wasp Asobara japonica is an ideal insect to investigate this because symbiotic and aposymbiotic strains are available: Wolbachia-infected Tokyo (TK) and noninfected Iriomote (IR) strains originally collected on the main island and southwest islands of Japan, respectively. We compared the oviposition behaviors of the two strains and found that TK strain females parasitized Drosophila melanogaster larvae more actively than the IR strain, especially during the first two days after eclosion. Removing Wolbachia from the TK strain wasps by treatment with tetracycline or rifampicin decreased their parasitism activity to the level of the IR strain. Morphological and behavioral analyses of both strain wasps showed that Wolbachia endosymbionts do not affect development of the host female reproductive tract and eggs, but do enhance host-searching ability of female wasps. These results suggest the possibility that Wolbachia endosymbionts may promote their diffusion and persistence in the host A. japonica population not only at least partly by parthenogenesis but also by enhancement of oviposition frequency of the host females. PMID:26492411

  15. Wolbachia endosymbionts induce neutrophil extracellular trap formation in human onchocerciasis

    PubMed Central

    Tamarozzi, Francesca; Turner, Joseph D.; Pionnier, Nicolas; Midgley, Angela; Guimaraes, Ana F.; Johnston, Kelly L.; Edwards, Steven W.; Taylor, Mark J.

    2016-01-01

    The endosymbiotic bacteria, Wolbachia, induce neutrophilic responses to the human helminth pathogen Onchocerca volvulus. The formation of Neutrophil Extracellular Traps (NETs), has been implicated in anti-microbial defence, but has not been identified in human helminth infection. Here, we demonstrate NETs formation in human onchocerciasis. Extracellular NETs and neutrophils were visualised around O. volvulus in nodules excised from untreated patients but not in nodules from patients treated with the anti-Wolbachia drug, doxycycline. Whole Wolbachia or microspheres coated with a synthetic Wolbachia lipopeptide (WoLP) of the major nematode Wolbachia TLR2/6 ligand, peptidoglycan associated lipoprotein, induced NETosis in human neutrophils in vitro. TLR6 dependency of Wolbachia and WoLP NETosis was demonstrated using purified neutrophils from TLR6 deficient mice. Thus, we demonstrate for the first time that NETosis occurs during natural human helminth infection and demonstrate a mechanism of NETosis induction via Wolbachia endobacteria and direct ligation of Wolbachia lipoprotein by neutrophil TLR2/6. PMID:27752109

  16. Comparative Genomics of a Parthenogenesis-Inducing Wolbachia Symbiont

    PubMed Central

    Lindsey, Amelia R. I.; Werren, John H.; Richards, Stephen; Stouthamer, Richard

    2016-01-01

    Wolbachia is an intracellular symbiont of invertebrates responsible for inducing a wide variety of phenotypes in its host. These host-Wolbachia relationships span the continuum from reproductive parasitism to obligate mutualism, and provide a unique system to study genomic changes associated with the evolution of symbiosis. We present the genome sequence from a parthenogenesis-inducing Wolbachia strain (wTpre) infecting the minute parasitoid wasp Trichogramma pretiosum. The wTpre genome is the most complete parthenogenesis-inducing Wolbachia genome available to date. We used comparative genomics across 16 Wolbachia strains, representing five supergroups, to identify a core Wolbachia genome of 496 sets of orthologous genes. Only 14 of these sets are unique to Wolbachia when compared to other bacteria from the Rickettsiales. We show that the B supergroup of Wolbachia, of which wTpre is a member, contains a significantly higher number of ankyrin repeat-containing genes than other supergroups. In the wTpre genome, there is evidence for truncation of the protein coding sequences in 20% of ORFs, mostly as a result of frameshift mutations. The wTpre strain represents a conversion from cytoplasmic incompatibility to a parthenogenesis-inducing lifestyle, and is required for reproduction in the Trichogramma host it infects. We hypothesize that the large number of coding frame truncations has accompanied the change in reproductive mode of the wTpre strain. PMID:27194801

  17. Mutualistic Wolbachia infection in Aedes albopictus: accelerating cytoplasmic drive.

    PubMed Central

    Dobson, Stephen L; Marsland, Eric J; Rattanadechakul, Wanchai

    2002-01-01

    Maternally inherited rickettsial symbionts of the genus Wolbachia occur commonly in arthropods, often behaving as reproductive parasites by manipulating host reproduction to enhance the vertical transmission of infections. One manipulation is cytoplasmic incompatibility (CI), which causes a significant reduction in brood hatch and promotes the spread of the maternally inherited Wolbachia infection into the host population (i.e., cytoplasmic drive). Here, we have examined a Wolbachia superinfection in the mosquito Aedes albopictus and found the infection to be associated with both cytoplasmic incompatibility and increased host fecundity. Relative to uninfected females, infected females live longer, produce more eggs, and have higher hatching rates in compatible crosses. A model describing Wolbachia infection dynamics predicts that increased fecundity will accelerate cytoplasmic drive rates. To test this hypothesis, we used population cages to examine the rate at which Wolbachia invades an uninfected Ae. albopictus population. The observed cytoplasmic drive rates were consistent with model predictions for a CI-inducing Wolbachia infection that increases host fecundity. We discuss the relevance of these results to both the evolution of Wolbachia symbioses and proposed applied strategies for the use of Wolbachia infections to drive desired transgenes through natural populations (i.e., population replacement strategies). PMID:11901124

  18. A host as an ecosystem: Wolbachia coping with environmental constraints.

    PubMed

    Sicard, Mathieu; Dittmer, Jessica; Grève, Pierre; Bouchon, Didier; Braquart-Varnier, Christine

    2014-12-01

    The Wolbachia are intracellular endosymbionts widely distributed among invertebrates. These primarily vertically transmitted α-proteobacteria have been intensively studied during the last decades because of their intriguing interactions with hosts, ranging from reproductive manipulations to mutualism. To optimize their vertical transmission from mother to offspring, the Wolbachia have developed fine-tuned strategies. However, the Wolbachia are not restricted to the female gonads and frequently exhibit wide intra-host distributions. This extensive colonization of somatic organs might be necessary for Wolbachia to develop their diverse extended phenotypes. From an endosymbiont's perspective, the within-host environment potentially presents different environmental constraints. Hence, the Wolbachia have to face different intracellular habitats, their host's immune system as well as other microorganisms co-occurring in the same host individual and sometimes even in the same cell. A means for the Wolbachia to protect themselves from these environmental constraints may be to live 'hidden' in vacuoles within host cells. In this review, we summarize the current knowledge regarding the extent of the Wolbachia pandemic and discuss the various environmental constraints these bacteria may have to face within their 'host ecosystem'. Finally, we identify new avenues for future research to better understand the complexity of Wolbachia's interactions with their intracellular environment.

  19. Macronutrients mediate the functional relationship between Drosophila and Wolbachia.

    PubMed

    Ponton, Fleur; Wilson, Kenneth; Holmes, Andrew; Raubenheimer, David; Robinson, Katie L; Simpson, Stephen J

    2015-02-07

    Wolbachia are maternally inherited bacterial endosymbionts that naturally infect a diverse array of arthropods. They are primarily known for their manipulation of host reproductive biology, and recently, infections with Wolbachia have been proposed as a new strategy for controlling insect vectors and subsequent human-transmissible diseases. Yet, Wolbachia abundance has been shown to vary greatly between individuals and the magnitude of the effects of infection on host life-history traits and protection against infection is correlated to within-host Wolbachia abundance. It is therefore essential to better understand the factors that modulate Wolbachia abundance and effects on host fitness. Nutrition is known to be one of the most important mediators of host-symbiont interactions. Here, we used nutritional geometry to quantify the role of macronutrients on insect-Wolbachia relationships in Drosophila melanogaster. Our results show fundamental interactions between diet composition, host diet selection, Wolbachia abundance and effects on host lifespan and fecundity. The results and methods described here open a new avenue in the study of insect-Wolbachia relationships and are of general interest to numerous research disciplines, ranging from nutrition and life-history theory to public health.

  20. The impact of host diet on Wolbachia titer in Drosophila.

    PubMed

    Serbus, Laura R; White, Pamela M; Silva, Jessica Pintado; Rabe, Amanda; Teixeira, Luis; Albertson, Roger; Sullivan, William

    2015-03-01

    While a number of studies have identified host factors that influence endosymbiont titer, little is known concerning environmental influences on titer. Here we examined nutrient impact on maternally transmitted Wolbachia endosymbionts in Drosophila. We demonstrate that Drosophila reared on sucrose- and yeast-enriched diets exhibit increased and reduced Wolbachia titers in oogenesis, respectively. The yeast-induced Wolbachia depletion is mediated in large part by the somatic TOR and insulin signaling pathways. Disrupting TORC1 with the small molecule rapamycin dramatically increases oocyte Wolbachia titer, whereas hyper-activating somatic TORC1 suppresses oocyte titer. Furthermore, genetic ablation of insulin-producing cells located in the Drosophila brain abolished the yeast impact on oocyte titer. Exposure to yeast-enriched diets altered Wolbachia nucleoid morphology in oogenesis. Furthermore, dietary yeast increased somatic Wolbachia titer overall, though not in the central nervous system. These findings highlight the interactions between Wolbachia and germline cells as strongly nutrient-sensitive, and implicate conserved host signaling pathways by which nutrients influence Wolbachia titer.

  1. Bacteria Endosymbiont, Wolbachia, Promotes Parasitism of Parasitoid Wasp Asobara japonica.

    PubMed

    Furihata, Shunsuke; Hirata, Makiko; Matsumoto, Hitoshi; Hayakawa, Yoichi

    2015-01-01

    Wolbachia is the most widespread endosymbiotic bacterium that manipulates reproduction of its arthropod hosts to enhance its own spread throughout host populations. Infection with Wolbachia causes complete parthenogenetic reproduction in many Hymenoptera, producing only female offspring. The mechanism of such reproductive manipulation by Wolbachia has been extensively studied. However, the effects of Wolbachia symbiosis on behavioral traits of the hosts are scarcely investigated. The parasitoid wasp Asobara japonica is an ideal insect to investigate this because symbiotic and aposymbiotic strains are available: Wolbachia-infected Tokyo (TK) and noninfected Iriomote (IR) strains originally collected on the main island and southwest islands of Japan, respectively. We compared the oviposition behaviors of the two strains and found that TK strain females parasitized Drosophila melanogaster larvae more actively than the IR strain, especially during the first two days after eclosion. Removing Wolbachia from the TK strain wasps by treatment with tetracycline or rifampicin decreased their parasitism activity to the level of the IR strain. Morphological and behavioral analyses of both strain wasps showed that Wolbachia endosymbionts do not affect development of the host female reproductive tract and eggs, but do enhance host-searching ability of female wasps. These results suggest the possibility that Wolbachia endosymbionts may promote their diffusion and persistence in the host A. japonica population not only at least partly by parthenogenesis but also by enhancement of oviposition frequency of the host females.

  2. Wolbachia distribution in selected beetle taxa characterized by PCR screens and MLST data.

    PubMed

    Sontowski, Rebekka; Bernhard, Detlef; Bleidorn, Christoph; Schlegel, Martin; Gerth, Michael

    2015-10-01

    Wolbachia (Alphaproteobacteria) is an inherited endosymbiont of arthropods and filarial nematodes and was reported to be widespread across insect taxa. While Wolbachia's effects on host biology are not understood from most of these hosts, known Wolbachia-induced phenotypes cover a spectrum from obligate beneficial mutualism to reproductive manipulations and pathogenicity. Interestingly, data on Wolbachia within the most species-rich order of arthropods, the Coleoptera (beetles), are scarce. Therefore, we screened 128 species from seven beetle families (Buprestidae, Hydraenidae, Dytiscidae, Hydrophilidae, Gyrinidae, Haliplidae, and Noteridae) for the presence of Wolbachia. Our data show that, contrary to previous estimations, Wolbachia frequencies in beetles (31% overall) are comparable to the ones in other insects. In addition, we used Wolbachia MLST data and host phylogeny to explore the evolutionary history of Wolbachia strains from Hydraenidae, an aquatic lineage of beetles. Our data suggest that Wolbachia from Hydraenidae might be largely host genus specific and that Wolbachia strain phylogeny is not independent to that of its hosts. As this contrasts with most terrestrial Wolbachia-arthropod systems, one potential conclusion is that aquatic lifestyle of hosts may result in Wolbachia distribution patterns distinct from those of terrestrial hosts. Our data thus provide both insights into Wolbachia distribution among beetles in general and a first glimpse of Wolbachia distribution patterns among aquatic host lineages.

  3. Rapid Sequential Spread of Two Wolbachia Variants in Drosophila simulans

    PubMed Central

    Kriesner, Peter; Hoffmann, Ary A.; Lee, Siu F.; Turelli, Michael; Weeks, Andrew R.

    2013-01-01

    The maternally inherited intracellular bacteria Wolbachia can manipulate host reproduction in various ways that foster frequency increases within and among host populations. Manipulations involving cytoplasmic incompatibility (CI), where matings between infected males and uninfected females produce non-viable embryos, are common in arthropods and produce a reproductive advantage for infected females. CI was associated with the spread of Wolbachia variant wRi in Californian populations of Drosophila simulans, which was interpreted as a bistable wave, in which local infection frequencies tend to increase only once the infection becomes sufficiently common to offset imperfect maternal transmission and infection costs. However, maternally inherited Wolbachia are expected to evolve towards mutualism, and they are known to increase host fitness by protecting against infectious microbes or increasing fecundity. We describe the sequential spread over approximately 20 years in natural populations of D. simulans on the east coast of Australia of two Wolbachia variants (wAu and wRi), only one of which causes significant CI, with wRi displacing wAu since 2004. Wolbachia and mtDNA frequency data and analyses suggest that these dynamics, as well as the earlier spread in California, are best understood as Fisherian waves of favourable variants, in which local spread tends to occur from arbitrarily low frequencies. We discuss implications for Wolbachia-host dynamics and coevolution and for applications of Wolbachia to disease control. PMID:24068927

  4. Modelling the spread of Wolbachia in spatially heterogeneous environments.

    PubMed

    Hancock, Penelope A; Godfray, H Charles J

    2012-11-07

    The endosymbiont Wolbachia infects a large number of insect species and is capable of rapid spread when introduced into a novel host population. The bacteria spread by manipulating their hosts' reproduction, and their dynamics are influenced by the demographic structure of the host population and patterns of contact between individuals. Reaction-diffusion models of the spatial spread of Wolbachia provide a simple analytical description of their spatial dynamics but do not account for significant details of host population dynamics. We develop a metapopulation model describing the spatial dynamics of Wolbachia in an age-structured host insect population regulated by juvenile density-dependent competition. The model produces similar dynamics to the reaction-diffusion model in the limiting case where the host's habitat quality is spatially homogeneous and Wolbachia has a small effect on host fitness. When habitat quality varies spatially, Wolbachia spread is usually much slower, and the conditions necessary for local invasion are strongly affected by immigration of insects from surrounding regions. Spread is most difficult when variation in habitat quality is spatially correlated. The results show that spatial variation in the density-dependent competition experienced by juvenile host insects can strongly affect the spread of Wolbachia infections, which is important to the use of Wolbachia to control insect vectors of human disease and other pests.

  5. Does a parthenogenesis-inducing Wolbachia induce vestigial cytoplasmic incompatibility?

    NASA Astrophysics Data System (ADS)

    Kraaijeveld, Ken; Reumer, Barbara M.; Mouton, Laurence; Kremer, Natacha; Vavre, Fabrice; van Alphen, Jacques J. M.

    2011-03-01

    Wolbachia is a maternally inherited bacterium that manipulates the reproduction of its host. Recent studies have shown that male-killing strains can induce cytoplasmic incompatibility (CI) when introgressed into a resistant host. Phylogenetic studies suggest that transitions between CI and other Wolbachia phenotypes have also occurred frequently, raising the possibility that latent CI may be widespread among Wolbachia. Here, we investigate whether a parthenogenesis-inducing Wolbachia strain can also induce CI. Parthenogenetic females of the parasitoid wasp Asobara japonica regularly produce a small number of males that may be either infected or not. Uninfected males were further obtained through removal of the Wolbachia using antibiotics and from a naturally uninfected strain. Uninfected females that had mated with infected males produced a slightly, but significantly more male-biased sex ratio than uninfected females that had mated with uninfected males. This effect was strongest in females that mated with males that had a relatively high Wolbachia titer. Quantitative PCR indicated that infected males did not show higher ratios of nuclear versus mitochondrial DNA content. Wolbachia therefore does not cause diploidization of cells in infected males. While these results are consistent with CI, other alternatives such as production of abnormal sperm by infected males cannot be completely ruled out. Overall, the effect was very small (9%), suggesting that if CI is involved it may have degenerated through the accumulation of mutations.

  6. Inactivation of Wolbachia Reveals Its Biological Roles in Whitefly Host

    PubMed Central

    Xue, Xia; Li, Shao-Jian; Ahmed, Muhammad Z.; De Barro, Paul J.; Ren, Shun-Xiang; Qiu, Bao-Li

    2012-01-01

    Background The whitefly Bemisia tabaci is cryptic species complex composed of numerous species. Individual species from the complex harbor a diversity of bacterial endosymbionts including Wolbachia. However, while Wolbachia is known to have a number of different roles, its role in B. tabaci is unclear. Here, the antibiotic rifampicin is used to selectively eliminate Wolbachia from B. tabaci so as to enable its roles in whitefly development and reproduction to be explored. The indirect effects of Wolbachia elimination on the biology of Encarsia bimaculata, a dominant parasitoid of B. tabaci in South China, were also investigated. Methodology/Principal Finding qRT-PCR and FISH were used to show that after 48 h exposure to 1.0 mg/ml rifampicin, Wolbachia was completely inactivated from B. tabaci Mediterranean (MED) without any significant impact on either the primary symbiont, Portiera aleyrodidarum or any of the other secondary endosymbionts present. For B. tabaci MED, Wolbachia was shown to be associated with decreased juvenile development time, increased likelihood that nymphs completed development, increased adult life span and increased percentage of female progeny. Inactivation was associated with a significant decrease in the body size of the 4th instar which leads us to speculate as to whether Wolbachia may have a nutrient supplementation role. The reduction in nymph body size has consequences for its parasitoid, E. bimaculata. The elimination of Wolbachia lead to a marked increase in the proportion of parasitoid eggs that completed their development, but the reduced size of the whitefly host was also associated with a significant reduction in the size of the emerging parasitoid adult and this was in turn associated with a marked reduction in adult parasitoid longevity. Conclusions/Significance Wolbachia increases the fitness of the whitefly host and provides some protection against parasitization. These observations add to our understanding of the roles

  7. The emerging role of Wolbachia species in heartworm disease.

    PubMed

    Frank, Kristen; Heald, R Dennis

    2010-04-01

    Heartworm disease was first recognized in dogs more than 100 years ago and is still prevalent among dogs and found in cats worldwide. The complications of heartworm disease can be devastating, and treatment carries risks. Wolbachia spp are gram-negative bacteria that infect filarial nematodes, including Dirofilaria immitis, and elicit an inflammatory response in cats and dogs. Antimicrobial therapy directed against these bacteria has resulted in decreased microfilarial loads, inhibition of the development of larval worms, female worm infertility, and reduced numbers of Wolbachia organisms. Antimicrobial therapy against Wolbachia spp may be useful in treating heartworm disease in cats and dogs, but further research is needed.

  8. [Genotypic Diversity of Wolbachia pipientis in Native and Invasive Harmonia axyridis Pall., 1773 (Coleoptera, Coccinellidae) Populations].

    PubMed

    Goryacheva, I I; Blekhman, A V; Andrianov, B V; Gorelova, T V; Zakharov, I A

    2015-08-01

    The distribution and variability of reproductive symbiotic Wolbachia pipientis bacteria were studied in seven native and six invasive H. axyridis populations. Wolbachia-infected individuals were found in two invasive and two native populations. We demonstrated for the first time an infection of invasive H. axyridis populations with Wolbachia. Two new molecular forms of Wolbachia were detected by a system of multilocus typing. The supergroup A Wolbachia was found for the first time in H. axyridis. The detected genotypic diversity of Wolbachia indicates repeated and independent infection events in the evolutionary past of H. axyridis.

  9. 150 Passenger Commercial Aircraft

    NASA Technical Reports Server (NTRS)

    Bucovsky, Adrian; Romli, Fairuz I.; Rupp, Jessica

    2002-01-01

    It has been projected that the need for a short-range mid-sized, aircraft is increasing. The future strategy to decrease long-haul flights will increase the demand for short-haul flights. Since passengers prefer to meet their destinations quickly, airlines will increase the frequency of flights, which will reduce the passenger load on the aircraft. If a point-to-point flight is not possible, passengers will prefer only a one-stop short connecting flight to their final destination. A 150-passenger aircraft is an ideal vehicle for these situations. It is mid-sized aircraft and has a range of 3000 nautical miles. This type of aircraft would market U.S. domestic flights or inter-European flight routes. The objective of the design of the 150-passenger aircraft is to minimize fuel consumption. The configuration of the aircraft must be optimized. This aircraft must meet CO2 and NOx emissions standards with minimal acquisition price and operating costs. This report contains all the work that has been performed for the completion of the design of a 150 passenger commercial aircraft. The methodology used is the Technology Identification, Evaluation, and Selection (TIES) developed at Georgia Tech Aerospace Systems Design laboratory (ASDL). This is an eight-step conceptual design process to evaluate the probability of meeting the design constraints. This methodology also allows for the evaluation of new technologies to be implemented into the design. The TIES process begins with defining the problem with a need established and a market targeted. With the customer requirements set and the target values established, a baseline concept is created. Next, the design space is explored to determine the feasibility and viability of the baseline aircraft configuration. If the design is neither feasible nor viable, new technologies can be implemented to open up the feasible design space and allow for a plausible solution. After the new technologies are identified, they must be evaluated

  10. Exploiting Intimate Relationships: Controlling Mosquito-Transmitted Disease with Wolbachia.

    PubMed

    Caragata, Eric P; Dutra, Heverton L C; Moreira, Luciano A

    2016-03-01

    Mosquito-transmitted diseases impose a growing burden on human health, and current control strategies have proven insufficient to stem the tide. The bacterium Wolbachia is a novel and promising form of control for mosquito-transmitted disease. It manipulates host biology, restricts infection with dengue and other pathogens, and alters host reproduction to promote rapid spread in the field. In this review, we examine how the intimate and diverse relationships formed between Wolbachia and their mosquito hosts can be exploited for disease control purposes. We consider these relationships in the context of recent developments, including successful field trials with Wolbachia-infected mosquitoes to combat dengue, and new Wolbachia infections in key malaria vectors, which have enhanced the disease control prospects of this unique bacterium.

  11. Temperature alters Plasmodium blocking by Wolbachia

    NASA Astrophysics Data System (ADS)

    Murdock, Courtney C.; Blanford, Simon; Hughes, Grant L.; Rasgon, Jason L.; Thomas, Matthew B.

    2014-02-01

    Very recently, the Asian malaria vector (Anopheles stephensi) was stably transinfected with the wAlbB strain of Wolbachia, inducing refractoriness to the human malaria parasite Plasmodium falciparum. However, conditions in the field can differ substantially from those in the laboratory. We use the rodent malaria P. yoelii, and somatically transinfected An. stephensi as a model system to investigate whether the transmission blocking potential of wAlbB is likely to be robust across different thermal environments. wAlbB reduced malaria parasite prevalence and oocyst intensity at 28°C. At 24°C there was no effect on prevalence but a marked increase in oocyst intensity. At 20°C, wAlbB had no effect on prevalence or intensity. Additionally, we identified a novel effect of wAlbB that resulted in reduced sporozoite development across temperatures, counterbalancing the oocyst enhancement at 24°C. Our results demonstrate complex effects of temperature on the Wolbachia-malaria interaction, and suggest the impacts of transinfection might vary across diverse environments.

  12. Association of Wolbachia with heartworm disease in cats and dogs.

    PubMed

    Dingman, Patricia; Levy, Julie K; Kramer, Laura H; Johnson, Calvin M; Lappin, Michael R; Greiner, Ellis C; Courtney, Charles H; Tucker, Sylvia J; Morchon, Rodrigo

    2010-05-28

    Although the presence of adult Dirofilaria immitis in the pulmonary arteries and its associated arteritis and thromboembolic disease can explain some of the manifestations of canine and feline heartworm disease, the cause of other findings remains unclear. Cats with D. immitis antibodies but lacking adult parasites in the pulmonary arteries frequently develop histological lesions of the airways, resulting in a condition termed Heartworm-Associated Respiratory Disease. All D. immitis parasites harbor Wolbachia pipientis bacteria and D. immitis-infected animals can have circulating Wolbachia antibodies and pro-inflammatory Wolbachia antigens (WSP) deposited in tissues. Little is known about the role that Wolbachia plays in lung disease of animals naturally infected with D. immitis. The purpose of this study was to determine the contribution of Wolbachia to the pathogenesis of natural heartworm disease in cats and dogs. We hypothesized that animals having sufficient Wolbachia burden to be detected in lung tissue by immunohistochemistry and/or polymerase chain reaction (PCR) would have more severe pulmonary disease than those with bacteria below the limits of detection. We further hypothesized that animals that were immunoreactive to pro-inflammatory WSP would have more severe pulmonary lesions than those that were seronegative for WSP antibodies. Blood and lung tissue samples were collected from cats and dogs representing three different D. immitis infection statuses: heartworm-free, heartworm-exposed, heartworm-infected. There was a positive but weak correlation between the magnitude of D. immitis antibody titers and WSP titers in cats (r=0.57, p<0.001) and in dogs (r=0.39, p<0.001). Pulmonary lesions were more common in HW-infected animals than in HW-free animals. Pulmonary arteriolar occlusion was more common in HW-infected cats (57%; p=0.003) than in HW-infected dogs (17%). Although pulmonary lesions were most common in HW-infected animals, there was no clear

  13. Wolbachia Modulates Lipid Metabolism in Aedes albopictus Mosquito Cells

    PubMed Central

    Molloy, Jennifer C.; Sommer, Ulf; Viant, Mark R.

    2016-01-01

    ABSTRACT Certain strains of the intracellular endosymbiont Wolbachia can strongly inhibit or block the transmission of viruses such as dengue virus (DENV) by Aedes mosquitoes, and the mechanisms responsible are still not well understood. Direct infusion and liquid chromatography-Fourier transform ion cyclotron resonance (FT-ICR) mass spectrometry-based lipidomics analyses were conducted using Aedes albopictus Aa23 cells that were infected with the wMel and wMelPop strains of Wolbachia in comparison to uninfected Aa23-T cells. Substantial shifts in the cellular lipid profile were apparent in the presence of Wolbachia. Most significantly, almost all sphingolipid classes were depleted, and some reductions in diacylglycerols and phosphatidylcholines were also observed. These lipid classes have previously been shown to be selectively enriched in DENV-infected mosquito cells, suggesting that Wolbachia may produce a cellular lipid environment that is antagonistic to viral replication. The data improve our understanding of the intracellular interactions between Wolbachia and mosquitoes. IMPORTANCE Mosquitoes transmit a variety of important viruses to humans, such as dengue virus and Zika virus. Certain strains of the intracellular bacterial genus called Wolbachia found in or introduced into mosquitoes can block the transmission of viruses, including dengue virus, but the mechanisms responsible are not well understood. We found substantial shifts in the cellular lipid profiles in the presence of these bacteria. Some lipid classes previously shown to be enriched in dengue virus-infected mosquito cells were depleted in the presence of Wolbachia, suggesting that Wolbachia may produce a cellular lipid environment that inhibits mosquito-borne viruses. PMID:26994075

  14. Child passenger safety.

    PubMed

    Durbin, Dennis R

    2011-04-01

    Despite significant reductions in the number of children killed in motor vehicle crashes over the past decade, crashes continue to be the leading cause of death for children 4 years and older. Therefore, the American Academy of Pediatrics continues to recommend inclusion of child passenger safety anticipatory guidance at every health-supervision visit. This technical report provides a summary of the evidence in support of 5 recommendations for best practices to optimize safety in passenger vehicles for children from birth through adolescence that all pediatricians should know and promote in their routine practice. These recommendations are presented in the revised policy statement on child passenger safety in the form of an algorithm that is intended to facilitate their implementation by pediatricians with their patients and families. The algorithm is designed to cover the majority of situations that pediatricians will encounter in practice. In addition, a summary of evidence on a number of additional issues that affect the safety of children in motor vehicles, including the proper use and installation of child restraints, exposure to air bags, travel in pickup trucks, children left in or around vehicles, and the importance of restraint laws, is provided. Finally, this technical report provides pediatricians with a number of resources for additional information to use when providing anticipatory guidance to families.

  15. Child passenger safety.

    PubMed

    Durbin, Dennis R

    2011-04-01

    Child passenger safety has dramatically evolved over the past decade; however, motor vehicle crashes continue to be the leading cause of death of children 4 years and older. This policy statement provides 4 evidence-based recommendations for best practices in the choice of a child restraint system to optimize safety in passenger vehicles for children from birth through adolescence: (1) rear-facing car safety seats for most infants up to 2 years of age; (2) forward-facing car safety seats for most children through 4 years of age; (3) belt-positioning booster seats for most children through 8 years of age; and (4) lap-and-shoulder seat belts for all who have outgrown booster seats. In addition, a fifth evidence-based recommendation is for all children younger than 13 years to ride in the rear seats of vehicles. It is important to note that every transition is associated with some decrease in protection; therefore, parents should be encouraged to delay these transitions for as long as possible. These recommendations are presented in the form of an algorithm that is intended to facilitate implementation of the recommendations by pediatricians to their patients and families and should cover most situations that pediatricians will encounter in practice. The American Academy of Pediatrics urges all pediatricians to know and promote these recommendations as part of child passenger safety anticipatory guidance at every health-supervision visit.

  16. Palaeosymbiosis Revealed by Genomic Fossils of Wolbachia in a Strongyloidean Nematode

    PubMed Central

    Koutsovoulos, Georgios; Makepeace, Benjamin; Tanya, Vincent N.; Blaxter, Mark

    2014-01-01

    Wolbachia are common endosymbionts of terrestrial arthropods, and are also found in nematodes: the animal-parasitic filaria, and the plant-parasite Radopholus similis. Lateral transfer of Wolbachia DNA to the host genome is common. We generated a draft genome sequence for the strongyloidean nematode parasite Dictyocaulus viviparus, the cattle lungworm. In the assembly, we identified nearly 1 Mb of sequence with similarity to Wolbachia. The fragments were unlikely to derive from a live Wolbachia infection: most were short, and the genes were disabled through inactivating mutations. Many fragments were co-assembled with definitively nematode-derived sequence. We found limited evidence of expression of the Wolbachia-derived genes. The D. viviparus Wolbachia genes were most similar to filarial strains and strains from the host-promiscuous clade F. We conclude that D. viviparus was infected by Wolbachia in the past, and that clade F-like symbionts may have been the source of filarial Wolbachia infections. PMID:24901418

  17. Palaeosymbiosis revealed by genomic fossils of Wolbachia in a strongyloidean nematode.

    PubMed

    Koutsovoulos, Georgios; Makepeace, Benjamin; Tanya, Vincent N; Blaxter, Mark

    2014-06-01

    Wolbachia are common endosymbionts of terrestrial arthropods, and are also found in nematodes: the animal-parasitic filaria, and the plant-parasite Radopholus similis. Lateral transfer of Wolbachia DNA to the host genome is common. We generated a draft genome sequence for the strongyloidean nematode parasite Dictyocaulus viviparus, the cattle lungworm. In the assembly, we identified nearly 1 Mb of sequence with similarity to Wolbachia. The fragments were unlikely to derive from a live Wolbachia infection: most were short, and the genes were disabled through inactivating mutations. Many fragments were co-assembled with definitively nematode-derived sequence. We found limited evidence of expression of the Wolbachia-derived genes. The D. viviparus Wolbachia genes were most similar to filarial strains and strains from the host-promiscuous clade F. We conclude that D. viviparus was infected by Wolbachia in the past, and that clade F-like symbionts may have been the source of filarial Wolbachia infections.

  18. Evidence of natural Wolbachia infections in field populations of Anopheles gambiae.

    PubMed

    Baldini, Francesco; Segata, Nicola; Pompon, Julien; Marcenac, Perrine; Shaw, W Robert; Dabiré, Roch K; Diabaté, Abdoulaye; Levashina, Elena A; Catteruccia, Flaminia

    2014-06-06

    Wolbachia are maternally transmitted intracellular bacteria that invade insect populations by manipulating their reproduction and immunity and thus limiting the spread of numerous human pathogens. Experimental Wolbachia infections can reduce Plasmodium numbers in Anopheles mosquitoes in the laboratory, however, natural Wolbachia infections in field anophelines have never been reported. Here we show evidence of Wolbachia infections in Anopheles gambiae in Burkina Faso, West Africa. Sequencing of the 16S rRNA gene identified Wolbachia sequences in both female and male germlines across two seasons, and determined that these sequences are vertically transmitted from mother to offspring. Whole-genome sequencing of positive samples suggests that the genetic material identified in An. gambiae belongs to a novel Wolbachia strain, related to but distinct from strains infecting other arthropods. The evidence of Wolbachia infections in natural Anopheles populations promotes further investigations on the possible use of natural Wolbachia-Anopheles associations to limit malaria transmission.

  19. Wolbachia infection density in populations of the Asian citrus psyllid (Hemiptera: Liviidae).

    PubMed

    Hoffmann, M; Coy, M R; Kingdom Gibbard, H N; Pelz-Stelinski, K S

    2014-10-01

    The symbiotic relationships between bacteria of the genus Wolbachia (order Rickettsiales) and their arthropod hosts are diverse and can range from mutualism to parasitism. Whereas effects of Wolbachia on host biology are well investigated, little is known about diversity and abundance of Wolbachia in their natural hosts. The phloem-feeding Asian citrus psyllid, Diaphorina citri (Kuwayama) (Hemiptera: Liviidae), is naturally infected with Wolbachia (wDi). In the current study, we calculated the within-host density of Wolbachia in Florida D. citri populations using quantitative polymerase chain reaction for detection of the Wolbachia outer surface protein gene, wsp. Gene quantities were normalized to the D. citri wingless gene (Wg) to estimate Wolbachia abundance in individual D. citri. Using this method, significant geographic differences in Wolbachia densities were detected among Florida D. citri populations, with higher infection levels occurring in male versus female hosts.

  20. Wolbachia Has Two Different Localization Patterns in Whitefly Bemisia tabaci AsiaII7 Species

    PubMed Central

    Shi, Peiqiong; He, Zhan; Li, Shaojian; An, Xuan; Lv, Ning; Ghanim, Murad; Cuthbertson, Andrew G. S.; Ren, Shun-Xiang

    2016-01-01

    The whitefly Bemisia tabaci is a cosmopolitan insect species complex that harbors the obligate primary symbiont Portiera aleyrodidarum and several facultative secondary symbionts including Wolbachia, which have diverse influences on the host biology. Here, for the first time, we revealed two different localization patterns of Wolbachia present in the immature and adult stages of B. tabaci AsiaII7 cryptic species. In the confined pattern, Wolbachia was restricted to the bacteriocytes, while in the scattered pattern Wolbachia localized in the bacteriocytes, haemolymph and other organs simultaneously. Our results further indicated that, the proportion of B. tabaci AsiaII7 individuals with scattered Wolbachia were significantly lower than that of confined Wolbachia, and the distribution patterns of Wolbachia were not associated with the developmental stage or sex of whitefly host. This study will provide a new insight into the various transmission routes of Wolbachia in different whitefly species. PMID:27611575

  1. PCR analysis for Wolbachia in human and canine Demodex mites.

    PubMed

    Borgo, Sibylle N; Sattler, Elke C; Hogardt, Michael; Adler, Kristin; Plewig, Gerd

    2009-10-01

    In many skin diseases such as Demodex folliculitis, rosacea- or steroid-induced rosacea Demodex mites are present in abundance and are at least partially held responsible for causing these disorders. Although it is known that these diseases respond well to tetracyclines, it is unclear if this is due to the antiinflammatory effects of the antibiotics or to an antibacterial effect on so far unknown bacteria within the Demodex mites. As in filariasis, where the response to doxycycline can be explained by the presence of Wolbachia within the filarial nematodes, this study was performed to see whether Wolbachia also use Demodex mites as their hosts. Human and canine Demodex mite samples were taken by skin scrapings and tested by PCR for the presence of Wolbachia DNA. Wolbachia pipientis DNA was used as positive control. In none of the DNA extracts, Wolbachia were detected showing no evidence for the presence of these bacteria in Demodex mites. The response of Demodex aggravated or Demodex caused diseases to tetracyclines seems not to be due to the presence of Wolbachia in Demodex mites in contrast to the results seen in filariasis.

  2. Breakdown of coevolution between symbiotic bacteria Wolbachia and their filarial hosts

    PubMed Central

    Lefoulon, Emilie; Makepeace, Benjamin L.; d’Haese, Cyrille; Uni, Shigehiko; Gavotte, Laurent

    2016-01-01

    Wolbachia is an alpha-proteobacterial symbiont widely distributed in arthropods. Since the identification of Wolbachia in certain animal-parasitic nematodes (the Onchocercidae or filariae), the relationship between arthropod and nematode Wolbachia has attracted great interest. The obligate symbiosis in filariae, which renders infected species susceptible to antibiotic chemotherapy, was held to be distinct from the Wolbachia-arthropod relationship, typified by reproductive parasitism. While co-evolutionary signatures in Wolbachia-arthropod symbioses are generally weak, reflecting horizontal transmission events, strict co-evolution between filariae and Wolbachia has been reported previously. However, the absence of close outgroups for phylogenetic studies prevented the determination of which host group originally acquired Wolbachia. Here, we present the largest co-phylogenetic analysis of Wolbachia in filariae performed to date including: (i) a screening and an updated phylogeny of Wolbachia; (ii) a co-phylogenetic analysis; and (iii) a hypothesis on the acquisition of Wolbachia infection. First, our results show a general overestimation of Wolbachia occurrence and support the hypothesis of an ancestral absence of infection in the nematode phylum. The accuracy of supergroup J is also underlined. Second, although a global pattern of coevolution remains, the signal is derived predominantly from filarial clades associated with Wolbachia in supergroups C and J. In other filarial clades, harbouring Wolbachia supergroups D and F, horizontal acquisitions and secondary losses are common. Finally, our results suggest that supergroup C is the basal Wolbachia clade within the Ecdysozoa. This hypothesis on the origin of Wolbachia would change drastically our understanding of Wolbachia evolution. PMID:27069790

  3. Wolbachia Endosymbionts Modify Drosophila Ovary Protein Levels in a Context-Dependent Manner

    PubMed Central

    Christensen, Steen; Pérez Dulzaides, Ricardo; Hedrick, Victoria E.; Momtaz, A. J. M. Zehadee; Nakayasu, Ernesto S.; Paul, Lake N.

    2016-01-01

    ABSTRACT Endosymbiosis is a unique form of interaction between organisms, with one organism dwelling inside the other. One of the most widespread endosymbionts is Wolbachia pipientis, a maternally transmitted bacterium carried by insects, crustaceans, mites, and filarial nematodes. Although candidate proteins that contribute to maternal transmission have been identified, the molecular basis for maternal Wolbachia transmission remains largely unknown. To investigate transmission-related processes in response to Wolbachia infection, ovarian proteomes were analyzed from Wolbachia-infected Drosophila melanogaster and D. simulans. Endogenous and variant host-strain combinations were investigated. Significant and differentially abundant ovarian proteins were detected, indicating substantial regulatory changes in response to Wolbachia. Variant Wolbachia strains were associated with a broader impact on the ovary proteome than endogenous Wolbachia strains. The D. melanogaster ovarian environment also exhibited a higher level of diversity of proteomic responses to Wolbachia than D. simulans. Overall, many Wolbachia-responsive ovarian proteins detected in this study were consistent with expectations from the experimental literature. This suggests that context-specific changes in protein abundance contribute to Wolbachia manipulation of transmission-related mechanisms in oogenesis. IMPORTANCE Millions of insect species naturally carry bacterial endosymbionts called Wolbachia. Wolbachia bacteria are transmitted by females to their offspring through a robust egg-loading mechanism. The molecular basis for Wolbachia transmission remains poorly understood at this time, however. This proteomic study identified specific fruit fly ovarian proteins as being upregulated or downregulated in response to Wolbachia infection. The majority of these protein responses correlated specifically with the type of host and Wolbachia strain involved. This work corroborates previously identified

  4. Costs and benefits of Wolbachia infection in immature Aedes albopictus depend upon sex and competition level.

    PubMed

    Gavotte, Laurent; Mercer, David R; Stoeckle, John J; Dobson, Stephen L

    2010-11-01

    Bacterial endosymbionts induce various effects on hosts and can dramatically impact host fitness and development. An example is provided by obligate, maternally-inherited Wolbachia, which infect a broad range of invertebrates. Wolbachia are capable of altering host reproduction, thereby promoting infection spread. Wolbachia also pose direct physiological costs and benefits to hosts, complicating their categorization as parasites or mutualists. This study examines for an effect of Wolbachia infection in intra-specific larval competition by Aedes albopictus mosquitoes, with the goal of examining for an impact of Wolbachia infection in mixed populations. Similar to prior work examining for an influence of Wolbachia infection on the fitness of A. albopictus in adults, the results presented here support the hypothesized impact of Wolbachia across all life stages, including immatures. The differential competitiveness of infected larvae detected in our experiments indicates that Wolbachia infected A. albopictus females are less competitive relative to uninfected females when competing under highly competitive conditions. In contrast, under low competitive pressures, infected females experience higher survivorship. Thus, Wolbachia infection shifts from parasitism to mutualism as a function of developmental conditions. Results are discussed in relation to the invasion and persistence of Wolbachia in A. albopictus populations. The results are important to the evolution of stable Wolbachia symbioses, including Wolbachia invasion of an uninfected population. The resulting infection dynamics that occur in an infected population are discussed.

  5. Diversification of Wolbachia endosymbiont in the Culex pipiens mosquito.

    PubMed

    Atyame, Célestine M; Delsuc, Frédéric; Pasteur, Nicole; Weill, Mylène; Duron, Olivier

    2011-10-01

    The α-proteobacteria Wolbachia are among the most common intracellular bacteria and have recently emerged as important drivers of arthropod biology. Wolbachia commonly act as reproductive parasites in arthropods by inducing cytoplasmic incompatibility (CI), a type of conditional sterility between hosts harboring incompatible infections. In this study, we examined the evolutionary histories of Wolbachia infections, known as wPip, in the common house mosquito Culex pipiens, which exhibits the greatest variation in CI crossing patterns observed in any insect. We first investigated a panel of 20 wPip strains for their genetic diversity through a multilocus scheme combining 13 Wolbachia genes. Because Wolbachia depend primarily on maternal transmission for spreading within arthropod populations, we also studied the variability in the coinherited Cx. pipiens mitochondria. In total, we identified 14 wPip haplotypes, which all share a monophyletic origin and clearly cluster into five distinct wPip groups. The diversity of Cx. pipiens mitochondria was extremely reduced, which is likely a consequence of cytoplasmic hitchhiking driven by a unique and recent Wolbachia invasion. Phylogenetic evidence indicates that wPip infections and mitochondrial DNA have codiverged through stable cotransmission within the cytoplasm and shows that a rapid diversification of wPip has occurred. The observed pattern demonstrates that a considerable degree of Wolbachia diversity can evolve within a single host species over short evolutionary periods. In addition, multiple signatures of recombination were found in most wPip genomic regions, leading us to conclude that the mosaic nature of wPip genomes may play a key role in their evolution.

  6. Molecular evidence of Wolbachia infection in natural populations of tropical odonates.

    PubMed

    Thipaksorn, Apisit; Jamnongluk, Wanwisa; Kittayapong, Pattamaporn

    2003-10-01

    Wolbachia are endosymbiotic bacteria that cause reproductive alterations in numerous arthropod species. Using a PCR-based method, we found that, out of 33 odonate species, four species were infected with Wolbachia. This finding represents the first record of Wolbachia infection in tropical odonates. Identical wsp gene sequences were found in the Wolbachia-infected common odonate species, Agriocnemis f. femina, collected from different locations in Thailand. The infection frequencies in several natural populations suggest that replacement of uninfected populations by Wolbachia-infected ones has recently occurred in this damselfly species.

  7. New Wolbachia supergroups detected in quill mites (Acari: Syringophilidae).

    PubMed

    Glowska, Eliza; Dragun-Damian, Anna; Dabert, Miroslawa; Gerth, Michael

    2015-03-01

    Wolbachia is the most abundant intracellular bacterial genus infecting a wide range of arthropods and filarial nematodes. Wolbachia have evolved parasitic, mutualistic and commensal relationships with their hosts but in arthropods generally act as reproductive parasites, inducing a wide range of phenotypic effects such as cytoplasmic incompatibility, parthenogenesis, feminization and male-killing. Up to now, the genus has been divided into 14 supergroups successively named A-O. Here, we describe two new Wolbachia supergroups from syringophilid mites (Acari: Cheyletoidea). These obligatory ectoparasites of birds inhabit the quills of feathers in many avian groups. The species of this family reproduce in a haplodiploid mode sensu arrhenotoky and are usually strongly female-biased. Based on the sequences of four protein-coding genes (ftsZ, gltA and groEL and coxA) and the 16S rRNA we identified strains of three Wolbachia supergroups (F and two distinct, yet undescribed ones) in five quill mite species. Our results suggest that in some cases the distribution of the bacteria can be better correlated with the mite's bird host rather than with mite taxonomy as such. The discovery of two new Wolbachia supergroups not only broadens the knowledge of the diversity of this bacterium but also raises questions about potential effects induced in quill mites and transmission mechanisms of the endosymbionts in this peculiar bacteria-quill mite-bird system.

  8. Human filarial Wolbachia lipopeptide directly activates human neutrophils in vitro.

    PubMed

    Tamarozzi, F; Wright, H L; Johnston, K L; Edwards, S W; Turner, J D; Taylor, M J

    2014-10-01

    The host inflammatory response to the Onchocerca volvulus endosymbiont, Wolbachia, is a major contributing factor in the development of chronic pathology in humans (onchocerciasis/river blindness). Recently, the toll-like pattern recognition receptor motif of the major inflammatory ligands of filarial Wolbachia, membrane-associated diacylated lipoproteins, was functionally defined in murine models of pathology, including mediation of neutrophil recruitment to the cornea. However, the extent to which human neutrophils can be activated in response to this Wolbachia pattern recognition motif is not known. Therefore, the responses of purified peripheral blood human neutrophils to a synthetic N-terminal diacylated lipopeptide (WoLP) of filarial Wolbachia peptidoglycan-associated lipoprotein (PAL) were characterized. WoLP exposure led to a dose-dependent activation of healthy, human neutrophils that included gross morphological alterations and modulation of surface expressed integrins involved in tethering, rolling and extravasation. WoLP exposure induced chemotaxis but not chemokinesis of neutrophils, and secretion of the major neutrophil chemokine, interleukin 8. WoLP also induced and primed the respiratory burst, and enhanced neutrophil survival by delay of apoptosis. These results indicate that the major inflammatory motif of filarial Wolbachia lipoproteins directly activates human neutrophils in vitro and promotes a molecular pathway by which human neutrophils are recruited to sites of Onchocerca parasitism.

  9. Wolbachia Infection in a Natural Parasitoid Wasp Population.

    PubMed

    Duplouy, Anne; Couchoux, Christelle; Hanski, Ilkka; van Nouhuys, Saskya

    2015-01-01

    The maternally transmitted bacterium Wolbachia pipientis is well known for spreading and persisting in insect populations through manipulation of the fitness of its host. Here, we identify three new Wolbachia pipientis strains, wHho, wHho2 and wHho3, infecting Hyposoter horticola, a specialist wasp parasitoid of the Glanville fritillary butterfly. The wHho strain (ST435) infects about 50% of the individuals in the Åland islands in Finland, with a different infection rate in the two mitochondrial (COI) haplotypes of the wasp. The vertical transmission rate of Wolbachia is imperfect, and lower in the haplotype with lower infection rate, suggesting a fitness trade-off. We found no association of the wHho infection with fecundity, longevity or dispersal ability of the parasitoid host. However, preliminary results convey spatial associations between Wolbachia infection, host mitochondrial haplotype and parasitism of H. horticola by its hyperparasitoid, Mesochorus cf. stigmaticus. We discuss the possibility that Wolbachia infection protects H. horticola against hyperparasitism.

  10. Flow cytometric evaluation of the intracellular bacterium, Wolbachia pipientis, in mosquito cells

    PubMed Central

    Fallon, Ann M

    2014-01-01

    Wolbachia is an obligate intracellular bacterium (Anaplasmataceae, Rickettisales) that occurs in arthropods and filarial worms, and spreads by vertical transmission in the oocyte cytoplasm. In insects, reproductive distortions associated with Wolbachia, such as cytoplasmic incompatibility in mosquitoes, have potential value for controlling pests, including species that transmit human, animal and plant diseases. Wolbachia strains that propagate as a persistent infection in insect cell lines provide an important resource for developing the genetic tools that will facilitate these applications. Here I describe conditions for flow cytometric evaluation of Wolbachia growth in persistently infected mosquito cells. Cytometry parameters were established using uninfected mosquito cells and Escherichia coli as a surrogate for Wolbachia, and quantitation was correlated with cell counts determined with a Coulter electronic cell counter and bacterial counts based on optical density. The protocol was validated by showing depletion of Wolbachia in medium containing tetracycline and rifampicin, and sensitivity of Wolbachia to treatment of host cells with paraquat, an oxidizing agent, and lumiflavin, an inhibitor of riboflavin uptake. The Wolbachia peak on the flow cytometry histogram was shown to contain Wolbachia by DNA analysis using the polymerase chain reaction, and by infection of naive recipient cells. This approach will streamline investigation of Wolbachia growth in insect cell lines and facilitate identification of culture conditions that select for Wolbachia-infected cells. PMID:25300665

  11. Depletion of host cell riboflavin reduces Wolbachia levels in cultured mosquito cells.

    PubMed

    Fallon, Ann M; Baldridge, Gerald D; Carroll, Elissa M; Kurtz, Cassandra M

    2014-09-01

    Wolbachia is an obligate intracellular alphaproteobacterium that occurs in arthropod and nematode hosts. Wolbachia presumably provides a fitness benefit to its hosts, but the basis for its retention and spread in host populations remains unclear. Wolbachia genomes retain biosynthetic pathways for some vitamins, and the possibility that these vitamins benefit host cells provides a potential means of selecting for Wolbachia-infected cell lines. To explore whether riboflavin produced by Wolbachia is available to its host cell, we established that growth of uninfected C7-10 mosquito cells decreases in riboflavin-depleted culture medium. A well-studied inhibitor of riboflavin uptake, lumiflavin, further inhibits growth of uninfected C7-10 cells with an LC50 of approximately 12 μg/ml. Growth of C/wStr1 mosquito cells, infected with Wolbachia from the planthopper, Laodelphax striatellus, was enhanced in medium containing low levels of lumiflavin, but Wolbachia levels decreased. Lumiflavin-enhanced growth thus resembled the improved growth that accompanies treatment with antibiotics that deplete Wolbachia, rather than a metabolic advantage provided by the Wolbachia infection. We used the polymerase chain reaction to validate the decrease in Wolbachia abundance and evaluated our results in the context of a proteomic analysis in which we detected nearly 800 wStr proteins. Our data indicate that Wolbachia converts riboflavin to FMN and FAD for its own metabolic needs, and does not provide a source of riboflavin for its host cell.

  12. Depletion of host cell riboflavin reduces Wolbachia levels in cultured mosquito cells

    PubMed Central

    Baldridge, Gerald D.; Carroll, Elissa M.; Kurtz, Cassandra M.

    2015-01-01

    Wolbachia is an obligate intracellular alphaproteobacterium that occurs in arthropod and nematode hosts. Wolbachia presumably provides a fitness benefit to its hosts, but the basis for its retention and spread in host populations remains unclear. Wolbachia genomes retain biosynthetic pathways for some vitamins, and the possibility that these vitamins benefit host cells provides a potential means of selecting for Wolbachia-infected cell lines. To explore whether riboflavin produced by Wolbachia is available to its host cell, we established that growth of uninfected C7–10 mosquito cells decreases in riboflavin-depleted culture medium. A well studied inhibitor of riboflavin uptake, lumiflavin, further inhibits growth of uninfected C7–10 cells with an LC50 of approximately 12 µg/ml. Growth of C/wStr1 mosquito cells, infected with Wolbachia from the planthopper, Laodelphax striatellus, was enhanced in medium containing low levels of lumiflavin, but Wolbachia levels decreased. Lumiflavin-enhanced growth thus resembled the improved growth that accompanies treatment with antibiotics that deplete Wolbachia, rather than a metabolic advantage provided by the Wolbachia infection. We used the polymerase chain reaction to validate the decrease in Wolbachia abundance and evaluated our results in the context of a proteomic analysis in which we detected nearly 800 wStr proteins. Our data indicate that Wolbachia converts riboflavin to FMN and FAD for its own metabolic needs, and does not provide a source of riboflavin for its host cell. PMID:24789726

  13. A Wolbachia-Sensitive Communication between Male and Female Pupae Controls Gamete Compatibility in Drosophila.

    PubMed

    Pontier, Stéphanie M; Schweisguth, François

    2015-09-21

    Gamete compatibility is fundamental to sexual reproduction. Wolbachia are maternally inherited endosymbiotic bacteria that manipulate gamete compatibility in many arthropod species. In Drosophila, the fertilization of uninfected eggs by sperm from Wolbachia-infected males often results in early developmental arrest. This gamete incompatibility is called cytoplasmic incompatibility (CI). CI is highest in young males, suggesting that Wolbachia affect sperm properties during male development. Here, we show that Wolbachia modulate testis development. Unexpectedly, this effect was associated with Wolbachia infection in females, not males. This raised the possibility that females influenced testis development by communicating with males prior to adulthood. Using a combinatorial rearing protocol, we provide evidence for such a female-to-male communication during metamorphosis. This communication involves the perception of female pheromones by male olfactory receptors. We found that this communication determines the compatibility range of sperm. Wolbachia interfere with this female-to-male communication through changes in female pheromone production. Strikingly, restoring this communication partially suppressed CI in Wolbachia-infected males. We further identified a reciprocal male-to-female communication at metamorphosis that restricts the compatibility range of female gametes. Wolbachia also perturb this communication by feminizing male pheromone production. Thus, Wolbachia broaden the compatibility range of eggs, promoting thereby the reproductive success of Wolbachia-infected females. We conclude that pheromone communication between pupae regulates gamete compatibility and is sensitive to Wolbachia in Drosophila.

  14. Bacteriophage WO Can Mediate Horizontal Gene Transfer in Endosymbiotic Wolbachia Genomes

    PubMed Central

    Wang, Guan H.; Sun, Bao F.; Xiong, Tuan L.; Wang, Yan K.; Murfin, Kristen E.; Xiao, Jin H.; Huang, Da W.

    2016-01-01

    Phage-mediated horizontal gene transfer (HGT) is common in free-living bacteria, and many transferred genes can play a significant role in their new bacterial hosts. However, there are few reports concerning phage-mediated HGT in endosymbionts (obligate intracellular bacteria within animal or plant hosts), such as Wolbachia. The Wolbachia-infecting temperate phage WO can actively shift among Wolbachia genomes and has the potential to mediate HGT between Wolbachia strains. In the present study, we extend previous findings by validating that the phage WO can mediate transfer of non-phage genes. To do so, we utilized bioinformatic, phylogenetic, and molecular analyses based on all sequenced Wolbachia and phage WO genomes. Our results show that the phage WO can mediate HGT between Wolbachia strains, regardless of whether the transferred genes originate from Wolbachia or other unrelated bacteria. PMID:27965627

  15. Detection of Wolbachia from field collected Aedes albopictus Skuse in Malaysia

    PubMed Central

    Afizah, A. Noor; Roziah, A.; Nazni, W.A.; Lee, H.L.

    2015-01-01

    Background & objectives: Wolbachia-based vector control strategies have been proposed as a mean to augment the existing measures for controlling dengue vector. Prior to utilizing Wolbachia in novel vector control strategies, it is crucial to understand the Wolbachia-mosquito interactions. Many studies have only focused on the prevalence of Wolbachia in female Aedes albopictus with lack of attention on Wolbachia infection on the male Ae. albopictus which also affects the effective expression of Wolbachia induced- cytoplasmic incompatibility (CI). In this study, field surveys were conducted to screen for the infection status of Wolbachia in female and male Ae. albopictus from various habitats including housing areas, islands and seashore. Methods: Adult Ae. albopictus (n=104) were collected using human landing catches and hand aspirator. Standard ovitraps were also set in the selected areas for five days and the larvae were identified to species level. All the collected Ae. albopictus were screened for the presence of Wolbachia using multiplex polymerase chain reaction (PCR) and gene sequencing of Wolbachia surface protein (wsp) gene. Results: A 100 per cent positivity of Wolbachia infection was observed for individual Ae. albopictus screened. For pooled mosquitoes, 73 of the 76 pools (female) and 83 of the 87 pools (male) were positive with Wolbachia infection. The wsp gene sequence of the Wolbachia strain isolated from individual and pooled mosquitoes showed a 100 per cent homology with Wolbachia sp. of Ae. albopictus isolated from various geographical regions. Phylogenetic analysis based on wsp gene fragments showed that the isolates were clustered into groups A and B, respectively. Interpretation & conclusions: The results indicated that Wolbachia infection was widespread in Ae. albopictus population both in female and male Ae. albopictus. All the infected females were superinfected with both A and B strains while the infected males showed a combination of

  16. Transfer of a parthenogenesis-inducing Wolbachia endosymbiont derived from Trichogramma dendrolimi into Trichogramma evanescens.

    PubMed

    Watanabe, Masaya; Kageyama, Daisuke; Miura, Kazuki

    2013-01-01

    Wolbachia, which are maternally transmitted endosymbionts, are considered to have moved horizontally between invertebrate hosts multiple times. However, it is not well understood how easily Wolbachia are transmitted horizontally between different hosts and how frequently horizontally-transmitted Wolbachia become established in their new hosts. We transferred a parthenogenesis-inducing Wolbachia endosymbiont derived from the parasitic wasp Trichogramma dendrolimi to Trichogramma evanescens. Specifically, Wolbachia was cultivated in a mosquito cell line and the Wolbachia-infected cells were microinjected into uninfected T. evanescens. Among 276 pupae inoculated with Wolbachia-infected cells, 65 adults emerged (G0). Diagnostic PCR demonstrated that 25 of 37 G0 females (68%) were Wolbachia-positive. Among isofemale lines established from G0 females, the proportions of infected lines were 80% (20 of 25) in G1 and 100% (18 of 18) in G2. In an isofemale line, infection was stably maintained for more than 10 generations. These results indicate invasion of Wolbachia into the germline of the recipient insect. Quantitative PCR demonstrated that the Wolbachia titer in the recipient host was significantly lower than that in the native host. The absence or very low number, if any, of parthenogenetically-reproducing individuals in the recipient host may be caused by the low Wolbachia titer. The Wolbachia titer in the recipients was lower in G11 than in G5, suggesting a decline in the density. Together with a previous report, our study may imply that Wolbachia in Trichogramma species are highly adapted to their hosts, which hinders robust expression of the Wolbachia phenotype in non-native host species.

  17. Modulation of host immunity and reproduction by horizontally acquired Wolbachia.

    PubMed

    Pigeault, Romain; Braquart-Varnier, Christine; Marcadé, Isabelle; Mappa, Gaëtan; Mottin, Elmina; Sicard, Mathieu

    2014-11-01

    The Wolbachia are symbiotic bacteria vertically transmitted from one host generation to another. However, a growing amount of data shows that horizontal transfers of Wolbachia also frequently occur within and between host species. The consequences of the arrival of new symbionts on host physiology can be studied by their experimental introduction in asymbiotic hosts. After experimental transfers of the eight major isopod Wolbachia strains in the isopod Porcellio dilatatus only two of them (wCon and wDil) were found to (1) have no pathogenic effect on the host and (2) be able to pass vertically to the host offspring. In the present work, we studied the influence of these two strains, able to complete an horizontal transfer, on immunity and reproduction of P. dilatatus at two stages of the transfer: (1) in recipient hosts that encounter the symbionts: to test the influence of symbiont when acquired during host life and (2) in vertically infected offspring: to test the influence of a symbiotic interaction occurring all lifelong. The impact of Wolbachia varied depending on the stage: there were clearer effects in vertically infected individuals than in those that acquired the symbionts during their lives. Moreover, the two Wolbachia strains showed contrasted effects: the strain wCon tended to reduce the reproductive investment but to maintain or increase immune parameters whilst wDil had positive effects on reproductive investment but decreased the investment in some immune parameters. These results suggest that horizontally acquisition of Wolbachia can influence the balance between host immune and reproductive traits.

  18. Comparison of Stable and Transient Wolbachia Infection Models in Aedes aegypti to Block Dengue and West Nile Viruses.

    PubMed

    Joubert, Dirk Albert; O'Neill, Scott L

    2017-01-01

    Pathogen replication and transmission in Wolbachia infected insects are currently studied using three Wolbachia infection systems: naturally infected Wolbachia hosts, hosts transinfected with Wolbachia (stably maintained and inherited infections) and hosts transiently infected with Wolbachia. All three systems have been used to test the effect of Wolbachia on mosquito transmitted pathogens such as dengue virus (DENV), West Nile virus (WNV) and Plasmodium. From these studies it is becoming increasingly clear that the interaction between a particular pathogen and Wolbachia is heavily influenced by the host-Wolbachia interaction and the model of infection. In particular, there is some evidence that under very specific conditions, Wolbachia can enhance pathogen infection in some hosts. In this study, we compared the effect of Wolbachia in two infection models (stable transinfected and transiently infected) on the replication, infection- and transmission rates of two flaviviruses, DENV and WNV (Kunjin strain). Our results indicate that Wolbachia had similar blocking effects in both stable and transient models of infection, however, the magnitude of the blocking effect was significantly lower in mosquitoes transiently infected with Wolbachia. More importantly, no evidence was found for any enhancement of either DENV or WNV (Kunjin strain) infection in Ae. aegypti infected with Wolbachia, supporting a role for Wolbachia as an effective and safe means for restricting transmission of these viruses.

  19. Comparison of Stable and Transient Wolbachia Infection Models in Aedes aegypti to Block Dengue and West Nile Viruses

    PubMed Central

    Joubert, Dirk Albert; O’Neill, Scott L.

    2017-01-01

    Pathogen replication and transmission in Wolbachia infected insects are currently studied using three Wolbachia infection systems: naturally infected Wolbachia hosts, hosts transinfected with Wolbachia (stably maintained and inherited infections) and hosts transiently infected with Wolbachia. All three systems have been used to test the effect of Wolbachia on mosquito transmitted pathogens such as dengue virus (DENV), West Nile virus (WNV) and Plasmodium. From these studies it is becoming increasingly clear that the interaction between a particular pathogen and Wolbachia is heavily influenced by the host-Wolbachia interaction and the model of infection. In particular, there is some evidence that under very specific conditions, Wolbachia can enhance pathogen infection in some hosts. In this study, we compared the effect of Wolbachia in two infection models (stable transinfected and transiently infected) on the replication, infection- and transmission rates of two flaviviruses, DENV and WNV (Kunjin strain). Our results indicate that Wolbachia had similar blocking effects in both stable and transient models of infection, however, the magnitude of the blocking effect was significantly lower in mosquitoes transiently infected with Wolbachia. More importantly, no evidence was found for any enhancement of either DENV or WNV (Kunjin strain) infection in Ae. aegypti infected with Wolbachia, supporting a role for Wolbachia as an effective and safe means for restricting transmission of these viruses. PMID:28052065

  20. Comparative genomics provides a timeframe for Wolbachia evolution and exposes a recent biotin synthesis operon transfer.

    PubMed

    Gerth, Michael; Bleidorn, Christoph

    2016-12-22

    The genus Wolbachia (Alphaproteobacteria) comprises the most abundant inherited intracellular bacteria(1). Despite their relevance as manipulators of human pathogen transmission(2) and arthropod reproduction(3), many aspects of their evolutionary history are not well understood(4). In arthropods, Wolbachia infections are typically transient on evolutionary timescales(5,6) and co-divergence between hosts and Wolbachia is supposedly rare. Consequently, much of our knowledge of Wolbachia genome evolution derives from very recently diverged strains, and a timescale for Wolbachia is lacking. Here, we investigated the genomes of four Wolbachia strains that have persisted within and co-diverged with their host lineage for ∼2 million years. Although the genomes showed very little evolutionary change on a nucleotide level, we found evidence for a recent lateral transfer of a complete biotin synthesis operon that has the potential to transform Wolbachia-host relationships(7). Furthermore, this evolutionary snapshot enabled us to calibrate the divergence times of the supergroup A and B Wolbachia lineages using genome-wide data sets and relaxed molecular clock models. We estimated the origin of Wolbachia supergroups A and B to be ∼200 million years ago (Ma), which is considerably older than previously appreciated. This age coincides with the diversification of many insect lineages(8) that represent most of Wolbachia's host spectrum.

  1. Transinfection: a method to investigate Wolbachia-host interactions and control arthropod-borne disease

    PubMed Central

    Hughes, Grant L.; Rasgon, Jason L.

    2014-01-01

    The bacterial endosymbiont Wolbachia manipulates arthropod host biology in numerous ways including sex ratio distortion and differential offspring survival. These bacteria infect a vast array of arthropods, some of which pose serious agricultural and human health threats. Wolbachia-mediated phenotypes such as cytoplasmic incompatibility and/or pathogen interference can be utilized for vector and disease control. However, many medically important vectors and important agricultural species are uninfected or are infected with strains of Wolbachia that do not elicit phenotypes desirable for disease or pest control. The ability to transfer strains of Wolbachia into new hosts (transinfection) can create novel Wolbachia-host associations. Transinfection has two primary benefits. First, Wolbachia-host interactions can be examined to tease apart the influence of the host and bacteria on phenotypes. Secondly, desirable phenotypes induced by Wolbachia in a particular insect can be transferred to another recipient host. This can allow for manipulation of insect populations that transmit pathogens or detrimentally affect agriculture. As such, transinfection is a valuable tool to explore Wolbachia biology and control arthropod-borne disease. This review summarizes what is currently known about Wolbachia transinfection methods and applications. We also provide a comprehensive list of published successful and unsuccessful Wolbachia transinfection attempts. PMID:24329998

  2. Passage of Wolbachia pipientis through mutant drosophila melanogaster induces phenotypic and genomic changes.

    PubMed

    Newton, Irene L G; Sheehan, Kathy B

    2015-02-01

    Wolbachia pipientis is a nearly ubiquitous, maternally transmitted bacterium that infects the germ line of insect hosts. Estimates are that Wolbachia infects 40 to 60% of insect species on the planet, making it one of the most prevalent infections on Earth. However, we know surprisingly little about the molecular mechanisms used by Wolbachia to infect its hosts. We passaged Wolbachia through normally restrictive Drosophila melanogaster hosts, bottlenecking Wolbachia through stochastic segregation while simultaneously selecting for mutants that could recolonize these previously restrictive hosts. Here, we show that Wolbachia alters its behavior when passaged through heterozygous mutant flies. After only three generations, Wolbachia was able to colonize the previously restrictive hosts at control titers. Additionally, the Wolbachia organisms passaged through heterozygous mutant D. melanogaster alter their pattern of tissue-specific Wsp protein production, suggesting a behavioral response to the host genotype. Using whole-genome resequencing, we identified the mutations accumulated by these lineages of Wolbachia and confirmed the existence and persistence of the mutations through clone library Sanger sequencing. Our results suggest that Wolbachia can quickly adapt to new host contexts, with genomic mutants arising after only two generations.

  3. Transinfection: a method to investigate Wolbachia-host interactions and control arthropod-borne disease.

    PubMed

    Hughes, G L; Rasgon, J L

    2014-04-01

    The bacterial endosymbiont Wolbachia manipulates arthropod host biology in numerous ways, including sex ratio distortion and differential offspring survival. These bacteria infect a vast array of arthropods, some of which pose serious agricultural and human health threats. Wolbachia-mediated phenotypes such as cytoplasmic incompatibility and/or pathogen interference can be used for vector and disease control; however, many medically important vectors and important agricultural species are uninfected or are infected with strains of Wolbachia that do not elicit phenotypes desirable for disease or pest control. The ability to transfer strains of Wolbachia into new hosts (transinfection) can create novel Wolbachia-host associations. Transinfection has two primary benefits. First, Wolbachia-host interactions can be examined to tease apart the influence of the host and bacteria on phenotypes. Second, desirable phenotypes induced by Wolbachia in a particular insect can be transferred to another recipient host. This can allow the manipulation of insect populations that transmit pathogens or detrimentally affect agriculture. As such, transinfection is a valuable tool to explore Wolbachia biology and control arthropod-borne disease. The present review summarizes what is currently known about Wolbachia transinfection methods and applications. We also provide a comprehensive list of published successful and unsuccessful Wolbachia transinfection attempts.

  4. Diversity and evolution of the Wolbachia endosymbionts of Bemisia (Hemiptera: Aleyrodidae) whiteflies

    PubMed Central

    Bing, Xiao-Li; Xia, Wen-Qiang; Gui, Jia-Dong; Yan, Gen-Hong; Wang, Xiao-Wei; Liu, Shu-Sheng

    2014-01-01

    Wolbachia is the most prevalent symbiont described in arthropods to date. Wolbachia can manipulate host reproduction, provide nutrition to insect hosts and protect insect hosts from pathogenic viruses. So far, 13 supergroups of Wolbachia have been identified. The whitefly Bemisia tabaci is a complex containing more than 28 morphologically indistinguishable cryptic species. Some cryptic species of this complex are invasive. In this study, we report a comprehensive survey of Wolbachia in B. tabaci and its relative B. afer from 1658 insects representing 54 populations across 13 provinces of China and one state of Australia. Based on the results of PCR or sequencing of the 16S rRNA gene, the overall rates of Wolbachia infection were 79.6% and 0.96% in the indigenous and invasive Bemisia whiteflies, respectively. We detected a new Wolbachia supergroup by sequencing five molecular marker genes including 16S rRNA, groEL, gltA, hcpA, and fbpA genes. Data showed that many protein-coding genes have limitations in detecting and classifying newly identified Wolbachia supergroups and thus raise a challenge to the known Wolbachia MLST standard analysis system. Besides, the other Wolbachia strains detected from whiteflies were clustered into supergroup B. Phylogenetic trees of whitefly mitochondrial cytochrome oxidase subunit I and Wolbachia multiple sequencing typing genes were not congruent. In addition, Wolbachia was also detected outside the special bacteriocytes in two cryptic species by fluorescence in situ hybridization, indicating the horizontal transmission of Wolbachia. Our results indicate that members of Wolbachia are far from well explored. PMID:25077022

  5. Tissue and Stage-Specific Distribution of Wolbachia in Brugia malayi

    PubMed Central

    Fischer, Kerstin; Beatty, Wandy L.; Jiang, Daojun; Weil, Gary J.; Fischer, Peter U.

    2011-01-01

    Background Most filarial parasite species contain Wolbachia, obligatory bacterial endosymbionts that are crucial for filarial development and reproduction. They are targets for alternative chemotherapy, but their role in the biology of filarial nematodes is not well understood. Light microscopy provides important information on morphology, localization and potential function of these bacteria. Surprisingly, immunohistology and in situ hybridization techniques have not been widely used to monitor Wolbachia distribution during the filarial life cycle. Methods/Principal Findings A monoclonal antibody directed against Wolbachia surface protein and in situ hybridization targeting Wolbachia 16S rRNA were used to monitor Wolbachia during the life cycle of B. malayi. In microfilariae and vector stage larvae only a few cells contain Wolbachia. In contrast, large numbers of Wolbachia were detected in the lateral chords of L4 larvae, but no endobacteria were detected in the genital primordium. In young adult worms (5 weeks p.i.), a massive expansion of Wolbachia was observed in the lateral chords adjacent to ovaries or testis, but no endobacteria were detected in the growth zone of the ovaries, uterus, the growth zone of the testis or the vas deferens. Confocal laser scanning and transmission electron microscopy showed that numerous Wolbachia are aligned towards the developing ovaries and single endobacteria were detected in the germline. In inseminated females (8 weeks p.i.) Wolbachia were observed in the ovaries, embryos and in decreasing numbers in the lateral chords. In young males Wolbachia were found in distinct zones of the testis and in large numbers in the lateral chords in the vicinity of testicular tissue but never in mature spermatids or spermatozoa. Conclusions Immunohistology and in situ hybridization show distinct tissue and stage specific distribution patterns for Wolbachia in B. malayi. Extensive multiplication of Wolbachia occurs in the lateral chords of L4

  6. Determinants of injuries in passenger vessel accidents.

    PubMed

    Yip, Tsz Leung; Jin, Di; Talley, Wayne K

    2015-09-01

    This paper investigates determinants of crew and passenger injuries in passenger vessel accidents. Crew and passenger injury equations are estimated for ferry, ocean cruise, and river cruise vessel accidents, utilizing detailed data of individual vessel accidents that were investigated by the U.S. Coast Guard during the time period 2001-2008. The estimation results provide empirical evidence (for the first time in the literature) that crew injuries are determinants of passenger injuries in passenger vessel accidents.

  7. Wolbachia and the insect immune system: what reactive oxygen species can tell us about the mechanisms of Wolbachia-host interactions.

    PubMed

    Zug, Roman; Hammerstein, Peter

    2015-01-01

    Wolbachia are intracellular bacteria that infect a vast range of arthropod species, making them one of the most prevalent endosymbionts in the world. Wolbachia's stunning evolutionary success is mostly due to their reproductive parasitism but also to mutualistic effects such as increased host fecundity or protection against pathogens. However, the mechanisms underlying Wolbachia phenotypes, both parasitic and mutualistic, are only poorly understood. Moreover, it is unclear how the insect immune system is involved in these phenotypes and why it is not more successful in eliminating the bacteria. Here we argue that reactive oxygen species (ROS) are likely to be key in elucidating these issues. ROS are essential players in the insect immune system, and Wolbachia infection can affect ROS levels in the host. Based on recent findings, we elaborate a hypothesis that considers the different effects of Wolbachia on the oxidative environment in novel vs. native hosts. We propose that newly introduced Wolbachia trigger an immune response and cause oxidative stress, whereas in coevolved symbioses, infection is not associated with oxidative stress, but rather with restored redox homeostasis. Redox homeostasis can be restored in different ways, depending on whether Wolbachia or the host is in charge. This hypothesis offers a mechanistic explanation for several of the observed Wolbachia phenotypes.

  8. Interspecific transfer of Wolbachia between two lepidopteran insects expressing cytoplasmic incompatibility: a Wolbachia variant naturally infecting Cadra cautella causes male killing in Ephestia kuehniella.

    PubMed Central

    Sasaki, Tetsuhiko; Kubo, Takeo; Ishikawa, Hajime

    2002-01-01

    Wolbachia is known as the causative agent of various reproductive alterations in arthropods. The almond moth Cadra cautella is doubly infected with A- and B-group Wolbachia and expresses complete cytoplasmic incompatibility (CI). The Mediterranean flour moth Ephestia kuehniella carries A-group Wolbachia and expresses partial CI. In the present study, the Wolbachia in C. cautella was transferred to E. kuehniella from which the original Wolbachia had been removed. We obtained transfected lines of three different infection states: single infection with A, single infection with B, and double infection with A and B. The doubly transfected lines and those transfected with only A produced exclusively female progeny. Two lines of evidence suggested that the sex ratio distortion was due to male killing. First, reduced egg hatch rate was observed. Second, removal of the Wolbachia from the transfected lines resulted in the recovery of a normal sex ratio of approximately 1:1. The occurrence of male killing following transfection showed that host factors influence the determination of the reproductive phenotype caused by Wolbachia. The transfected E. kuehniella males carrying exclusively B-group Wolbachia expressed partial incompatibility when crossed with the uninfected females. In addition, the transfected lines were bidirectionally incompatible with the naturally infected strain, which was the first demonstration of bidirectional CI in a lepidopteran. PMID:12454075

  9. Assessing the epidemiological effect of wolbachia for dengue control.

    PubMed

    Lambrechts, Louis; Ferguson, Neil M; Harris, Eva; Holmes, Edward C; McGraw, Elizabeth A; O'Neill, Scott L; Ooi, Eng E; Ritchie, Scott A; Ryan, Peter A; Scott, Thomas W; Simmons, Cameron P; Weaver, Scott C

    2015-07-01

    Dengue viruses cause more human morbidity and mortality than any other arthropod-borne virus. Dengue prevention relies mainly on vector control; however, the failure of traditional methods has promoted the development of novel entomological approaches. Although use of the intracellular bacterium wolbachia to control mosquito populations was proposed 50 years ago, only in the past decade has its use as a potential agent of dengue control gained substantial interest. Here, we review evidence that supports a practical approach for dengue reduction through field release of wolbachia-infected mosquitoes and discuss the additional studies that have to be done before the strategy can be validated and implemented. A crucial next step is to assess the efficacy of wolbachia in reducing dengue virus transmission. We argue that a cluster randomised trial is at this time premature because choice of wolbachia strain for release and deployment strategies are still being optimised. We therefore present a pragmatic approach to acquiring preliminary evidence of efficacy through various complementary methods including a prospective cohort study, a geographical cluster investigation, virus phylogenetic analysis, virus surveillance in mosquitoes, and vector competence assays. This multipronged approach could provide valuable intermediate evidence of efficacy to justify a future cluster randomised trial.

  10. Inhibition of Zika virus by Wolbachia in Aedes aegypti

    PubMed Central

    Caragata, Eric Pearce; Dutra, Heverton Leandro Carneiro; Moreira, Luciano Andrade

    2016-01-01

    Through association with cases of microcephaly in 2015, Zika virus (ZIKV) has transitioned from a relatively unknown mosquito-transmitted pathogen to a global health emergency, emphasizing the need to improve existing mosquito control programs to prevent future disease outbreaks. The response to Zika must involve a paradigm shift from traditional to novel methods of mosquito control, and according to the World Health Organization should incorporate the release of mosquitoes infected with the bacterial endosymbiont Wolbachia pipientis. In our recent paper [Dutra, HLC et al., Cell Host & Microbe 2016] we investigated the potential of Wolbachia infections in Aedes aegypti to restrict infection and transmission of Zika virus recently isolated in Brazil. Wolbachia is now well known for its ability to block or reduce infection with a variety of pathogens in different mosquito species including the dengue (DENV), yellow fever, and chikungunya viruses, and malaria-causing Plasmodium, and consequently has great potential to control mosquito-transmitted diseases across the globe. Our results demonstrated that the wMel Wolbachia strain in Brazilian Ae. aegypti is a strong inhibitor of ZIKV infection, and furthermore appears to prevent transmission of infectious viral particles in mosquito saliva, which highlights the bacterium’s suitability for more widespread use in Zika control. PMID:28357366

  11. Wolbachia strains for disease control: ecological and evolutionary considerations

    PubMed Central

    Hoffmann, Ary A; Ross, Perran A; Rašić, Gordana

    2015-01-01

    Wolbachia are endosymbionts found in many insects with the potential to suppress vectorborne diseases, particularly through interfering with pathogen transmission. Wolbachia strains are highly variable in their effects on hosts, raising the issue of which attributes should be selected to ensure that the best strains are developed for disease control. This depends on their ability to suppress viral transmission, invade host populations, persist without loss of viral suppression and not interfere with other control strategies. The potential to achieve these objectives is likely to involve evolutionary constraints; viral suppression may be limited by the ability of infections to spread due to deleterious host fitness effects. However, there are exceptions to these patterns in both natural infections and in novel associations generated following interspecific transfer, suggesting that pathogen blockage, deleterious fitness effects and changes to reproductive biology might be at least partly decoupled to achieve ideal infection attributes. The stability of introduced Wolbachia and its effects on viral transmission remain unclear, but rapid evolutionary changes seem unlikely. Although deliberate transfers of Wolbachia across species remain particularly challenging, the availability of strains with desirable attributes should be expanded, taking advantage of the diversity available across thousands of strains in natural populations. PMID:26366194

  12. The Wolbachia endosymbiont as an anti-filarial nematode target

    PubMed Central

    Taylor, Mark J.; Foster, Jeremy M.

    2010-01-01

    Human disease caused by parasitic filarial nematodes is a major cause of global morbidity. The parasites are transmitted by arthropod intermediate hosts and are responsible for lymphatic filariasis (elephantiasis) or onchocerciasis (river blindness). Within these filarial parasites are intracellular alpha-proteobacteria, Wolbachia, that were first observed almost 30 years ago. The obligate endosymbiont has been recognized as a target for anti-filarial nematode chemotherapy as evidenced by the loss of worm fertility and viability upon antibiotic treatment in an extensive series of human trials. While current treatments with doxycycline and rifampicin are not practical for widespread use due to the length of required treatments and contraindications, anti-Wolbachia targeting nevertheless appears a promising alternative for filariasis control in situations where current programmatic strategies fail or are unable to be delivered and it provides a superior efficacy for individual therapy. The mechanisms that underlie the symbiotic relationship between Wolbachia and its nematode hosts remain elusive. Comparative genomics, bioinfomatic and experimental analyses have identified a number of potential interactions, which may be drug targets. One candidate is de novo heme biosynthesis, due to its absence in the genome sequence of the host nematode, Brugia malayi, but presence in Wolbachia and its potential roles in worm biology. We describe this and several additional candidate targets, as well as our approaches for understanding the nature of the host-symbiont relationship. PMID:20730111

  13. Effects of Wolbachia on fitness of Culex quinquefasciatus (Diptera; Culicidae).

    PubMed

    Almeida, Fábio de; Moura, Alexandre S; Cardoso, André F; Winter, Carlos E; Bijovsky, A Tania; Suesdek, Lincoln

    2011-12-01

    Wolbachia are α-proteobacteria that were first reported in Culex pipiens mosquitoes early in the twentieth century. Since then, the effect of Wolbachia on their host's reproduction has drawn attention and has been increasingly investigated. Given the extreme complexity of this interaction, new study cases are welcomed to enhance its understanding. The present work addressed the influence of Wolbachia on Cx. quinquefasciatus, the cosmopolitan member of the Cx. pipiens complex. Samples of a Cx. quinquefasciatus colony (wPip(+)) originated from individuals naturally infected by Wolbachiapipientis B strain, were cured with tetracycline, yielding a Wolbachia-free colony (wPip(-)). Both the presence of bacteria and the efficiency of bacterial elimination were checked by PCR of the wsp gene. Total reproductive unidirectional incompatibility occurred when wPip(-) females were crossed with wPip(+) males, whereas the other three types of reciprocal crosses were viable. Reproductive aspects were also comparatively evaluated between colonies. Concerning oviposition time during the first gonotrophic cycle, wPip(+) females developed and laid eggs earlier than did wPip(-) females. Reproductive fitness was higher among wPip(-) than wPip(+) females regarding the following parameters: fertility: egg rafts/fed females; fecundity: eggs/raft, and viability: larvae/eggs. Conversely, longevity of wPip(-) females was lower. Summarising, although the infected mosquitoes have the advantage of a higher longevity, they have lower reproductive fitness. Our results are partly distinct from all other reports on Aedes and Culex mosquitoes previously published.

  14. Detection of Wolbachia in Aedes albopictus and Their Effects on Chikungunya Virus

    PubMed Central

    Ahmad, Noor Afizah; Vythilingam, Indra; Lim, Yvonne A. L.; Zabari, Nur Zatil Aqmar M.; Lee, Han Lim

    2017-01-01

    Wolbachia-based vector control strategies have been proposed as a means to augment the currently existing measures for controlling dengue and chikungunya vectors. Prior to utilizing Wolbachia as a novel vector control strategy, it is crucial to understand the Wolbachia–mosquito interactions. In this study, field surveys were conducted to screen for the infection status of Wolbachia in field-collected Aedes albopictus. The effects of Wolbachia in its native host toward the replication and dissemination of chikungunya virus (CHIKV) was also studied. The prevalence of Wolbachia-infected field-collected Ae. albopictus was estimated to be 98.6% (N = 142) for females and 95.1% (N = 102) for males in the population studied. The Ae. albopictus were naturally infected with both wAlbA and wAlbB strains. We also found that the native Wolbachia has no impact on CHIKV infection and minimal effect on CHIKV dissemination to secondary organs. PMID:27920393

  15. Ultrastructural and molecular identification of a Wolbachia endosymbiont in a spider, Nephila clavata.

    PubMed

    Oh, H W; Kim, M G; Shin, S W; Bae, K S; Ahn, Y J; Park, H Y

    2000-10-01

    Wolbachia-like bacteria were observed in the egg cells of golden orb-weaving spider, Nephila clavata, by means of transmission electron microscopy. The bacteria exhibited the typical morphology of Wolbachia, including three enveloping membranes. Based on the amplification and sequencing of partial 16S rDNA and ftsZ gene, the bacteria were identified as Wolbachia, intracellular, transovarially inherited alpha-proteobacteria in invertebrates. Phylogenetic analysis based on 16S rDNA and ftsZ gene sequences invariably indicated that the intracellular bacteria from N. clavata belonged to group A Wolbachia, which were found only from insects. Clustering of Wolbachia from N. clavata with group A Wolbachia indicates that the bacteria were probably transferred horizontally between insects and the spider.

  16. Molecular evidence of Wolbachia endosymbiosis in Mansonella perstans in Gabon, Central Africa.

    PubMed

    Gehringer, Christian; Kreidenweiss, Andrea; Flamen, Arnaud; Antony, Justin S; Grobusch, Martin P; Bélard, Sabine

    2014-11-15

    The discovery of obligatory intracellular bacteria of the genus Wolbachia in filariae infecting humans led to the use of antibiotics as a potent treatment option. Mansonella perstans is the cause of the second most prevalent filariasis in Gabon, but so far reports on the presence of Wolbachia in this nematode have been inconsistent. We report on the presence of Wolbachia in M. perstans in patients from Gabon, which we identified using polymerase chain reaction (PCR) with primer sets specific for 16S rDNA and ftsZ. Sequence analysis revealed a single consensus sequence, which could be phylogenetically assigned to Wolbachia of the supergroup F. Wolbachia could only be identified in 5 of 14 or 7 of 14 cases, depending on the investigated gene; detection of Wolbachia was associated with higher-level filaremia. Before generalizing the use of antibiotics for mansonellosis, further clarification of the obligatory nature of the endosymbiosis in this nematode is needed.

  17. Pathogenicity of life-shortening Wolbachia in Aedes albopictus after transfer from Drosophila melanogaster.

    PubMed

    Suh, Eunho; Mercer, David R; Fu, Yuqing; Dobson, Stephen L

    2009-12-01

    Maternally inherited Wolbachia bacteria have evolved mechanisms to manipulate the reproduction of their invertebrate hosts, promoting infection spread. A high fitness cost to the host is maladaptive for obligate endosymbionts, and prior studies show rapid selection of new Wolbachia associations toward commensal or mutualistic symbioses. Here, wMelPop Wolbachia is transferred from Drosophila melanogaster into the mosquito Aedes albopictus. Characterization of the resulting strain provides an extreme example of Wolbachia as a pathogen. In addition to reduced longevity and fecundity, abnormally high Wolbachia density is associated with embryonic mortality that masks the typical pattern of cytoplasmic incompatibility. The results are consistent with earlier reports that show unpredictable shifts in the Wolbachia phenotype after interspecific transfer, which can complicate proposed strategies to modify the age structure of medically important vector populations.

  18. A Study of the Educationally Influential Physician.

    ERIC Educational Resources Information Center

    Kaufman, David M.; Ryan, Kurt; Hodder, Ian

    1999-01-01

    A survey of 172 family doctors found that they approached educationally influential (EI) physicians they knew through their hospitals; only 20% used e-mail and 40% the Internet for medical information; EI physicians helped extend their knowledge and validate innovations found in the literature; and health care reform was negatively affecting…

  19. Evolutionary origin of insect–Wolbachia nutritional mutualism

    PubMed Central

    Nikoh, Naruo; Hosokawa, Takahiro; Moriyama, Minoru; Oshima, Kenshiro; Hattori, Masahira; Fukatsu, Takema

    2014-01-01

    Obligate insect–bacterium nutritional mutualism is among the most sophisticated forms of symbiosis, wherein the host and the symbiont are integrated into a coherent biological entity and unable to survive without the partnership. Originally, however, such obligate symbiotic bacteria must have been derived from free-living bacteria. How highly specialized obligate mutualisms have arisen from less specialized associations is of interest. Here we address this evolutionary issue by focusing on an exceptional insect–Wolbachia nutritional mutualism. Although Wolbachia endosymbionts are ubiquitously found in diverse insects and generally regarded as facultative/parasitic associates for their insect hosts, a Wolbachia strain associated with the bedbug Cimex lectularius, designated as wCle, was shown to be essential for host’s growth and reproduction via provisioning of B vitamins. We determined the 1,250,060-bp genome of wCle, which was generally similar to the genomes of insect-associated facultative Wolbachia strains, except for the presence of an operon encoding the complete biotin synthetic pathway that was acquired via lateral gene transfer presumably from a coinfecting endosymbiont Cardinium or Rickettsia. Nutritional and physiological experiments, in which wCle-infected and wCle-cured bedbugs of the same genetic background were fed on B-vitamin–manipulated blood meals via an artificial feeding system, demonstrated that wCle certainly synthesizes biotin, and the wCle-provisioned biotin significantly contributes to the host fitness. These findings strongly suggest that acquisition of a single gene cluster consisting of biotin synthesis genes underlies the bedbug–Wolbachia nutritional mutualism, uncovering an evolutionary transition from facultative symbiosis to obligate mutualism facilitated by lateral gene transfer in an endosymbiont lineage. PMID:24982177

  20. Evolutionary origin of insect-Wolbachia nutritional mutualism.

    PubMed

    Nikoh, Naruo; Hosokawa, Takahiro; Moriyama, Minoru; Oshima, Kenshiro; Hattori, Masahira; Fukatsu, Takema

    2014-07-15

    Obligate insect-bacterium nutritional mutualism is among the most sophisticated forms of symbiosis, wherein the host and the symbiont are integrated into a coherent biological entity and unable to survive without the partnership. Originally, however, such obligate symbiotic bacteria must have been derived from free-living bacteria. How highly specialized obligate mutualisms have arisen from less specialized associations is of interest. Here we address this evolutionary issue by focusing on an exceptional insect-Wolbachia nutritional mutualism. Although Wolbachia endosymbionts are ubiquitously found in diverse insects and generally regarded as facultative/parasitic associates for their insect hosts, a Wolbachia strain associated with the bedbug Cimex lectularius, designated as wCle, was shown to be essential for host's growth and reproduction via provisioning of B vitamins. We determined the 1,250,060-bp genome of wCle, which was generally similar to the genomes of insect-associated facultative Wolbachia strains, except for the presence of an operon encoding the complete biotin synthetic pathway that was acquired via lateral gene transfer presumably from a coinfecting endosymbiont Cardinium or Rickettsia. Nutritional and physiological experiments, in which wCle-infected and wCle-cured bedbugs of the same genetic background were fed on B-vitamin-manipulated blood meals via an artificial feeding system, demonstrated that wCle certainly synthesizes biotin, and the wCle-provisioned biotin significantly contributes to the host fitness. These findings strongly suggest that acquisition of a single gene cluster consisting of biotin synthesis genes underlies the bedbug-Wolbachia nutritional mutualism, uncovering an evolutionary transition from facultative symbiosis to obligate mutualism facilitated by lateral gene transfer in an endosymbiont lineage.

  1. Sequences of Wolbachia wsp genes reveal multiple infection of individual northern corn rootworms (Diabrotica barberi) by several Wolbachia strains

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Northern corn rootworm (Diabrotica barberi)(NCR) populations in the USA are infected with at least 4 strains of the endosymbiont, Wolbachia. NCR from eastern Illinois to Pennsylvania appear to harbor at least 4 different strains designated wBar1, wBar3, wBar4, and wBar5. NCR from central Illinois ...

  2. Wolbachia Endobacteria in Natural Populations of Culex pipiens of Iran and Its Phylogenetic Congruence

    PubMed Central

    Karami, Mohsen; Moosa-Kazemi, Seyed Hassan; Oshaghi, Mohammad Ali; Vatandoost, Hasan; Sedaghat, Mohammad Mehdi; Rajabnia, Ramazan; Hosseini, Mostafa; Maleki-Ravasan, Naseh; Yahyapour, Yousef; Ferdosi-Shahandashti, Elaheh

    2016-01-01

    Background: Wolbachia are common intracellular bacteria that infect different groups of arthropods including mosquitoes. These bacteria modify host biology and may induce feminization, parthenogenesis, male killing and cytoplasmic incompatibility (CI). Recently Wolbachia is being nominated as a bio-agent and paratransgenic candidate to control mosquito borne diseases. Methods: Here we report the results of a survey for presence, frequency, and phylogenetic congruence of these endosymbiont bacteria in Culex pipiens populations in Northern, Central, and Southern parts of Iran using nested-PCR amplification of wsp gene. Results: Wolbachia DNA were found in 227 (87.3%) out of 260 wild-caught mosquitoes. The rate of infection in adult females ranged from 61.5% to 100%, while in males were from 80% to 100%. The Blast search and phylogenetic analysis of the wsp gene sequence revealed that the Wolbachia strain from Iranian Cx. pipiens was identical to the Wolbachia strains of supergroup B previously reported in members of the Cx. pipiens complex. They had also identical sequence homology with the Wolbachia strains from a group of distinct arthropods including lepidopteran, wasps, flies, damselfly, thrips, and mites from remote geographical areas of the world. Conclusion: It is suggested that Wolbachia strains horizontally transfer between unrelated host organisms over evolutionary time. Also results of this study indicates that Wolbachia infections were highly prevalent infecting all Cx. pipiens populations throughout the country, however further study needs to define Wolbachia inter-population reproductive incompatibility pattern and its usefulness as a bio-agent control measure. PMID:27308293

  3. Wolbachia enhance Drosophila stem cell proliferation and target the germline stem cell niche.

    PubMed

    Fast, Eva M; Toomey, Michelle E; Panaram, Kanchana; Desjardins, Danielle; Kolaczyk, Eric D; Frydman, Horacio M

    2011-11-18

    Wolbachia are widespread maternally transmitted intracellular bacteria that infect most insect species and are able to alter the reproduction of innumerous hosts. The cellular bases of these alterations remain largely unknown. Here, we report that Drosophila mauritiana infected with a native Wolbachia wMau strain produces about four times more eggs than the noninfected counterpart. Wolbachia infection leads to an increase in the mitotic activity of germline stem cells (GSCs), as well as a decrease in programmed cell death in the germarium. Our results suggest that up-regulation of GSC division is mediated by a tropism of Wolbachia for the GSC niche, the cellular microenvironment that supports GSCs.

  4. Bad guys turned nice? A critical assessment of Wolbachia mutualisms in arthropod hosts.

    PubMed

    Zug, Roman; Hammerstein, Peter

    2015-02-01

    Wolbachia are the most abundant bacterial endosymbionts among arthropods. Although maternally inherited, they do not conform to the widespread view that vertical transmission inevitably selects for beneficial symbionts. Instead, Wolbachia are notorious for their reproductive parasitism which, although lowering host fitness, ensures their spread. However, even for reproductive parasites it can pay to enhance host fitness. Indeed, there is a recent upsurge of reports on Wolbachia-associated fitness benefits. Therefore, the question arises how such instances of mutualism are related to the phenotypes of reproductive parasitism. Here, we review the evidence of Wolbachia mutualisms in arthropods, including both facultative and obligate relationships, and critically assess their biological relevance. Although many studies report anti-pathogenic effects of Wolbachia, few actually prove these effects to be relevant to field conditions. We further show that Wolbachia frequently have beneficial and detrimental effects at the same time, and that reproductive manipulations and obligate mutualisms may share common mechanisms. These findings undermine the idea of a clear-cut distinction between Wolbachia mutualism and parasitism. In general, both facultative and obligate mutualisms can have a strong, and sometimes unforeseen, impact on the ecology and evolution of Wolbachia and their arthropod hosts. Acknowledging this mutualistic potential might be the key to a better understanding of some unresolved issues in the study of Wolbachia-host interactions.

  5. Quantitative Proteomic Analyses of Molecular Mechanisms Associated with Cytoplasmic Incompatibility in Drosophila melanogaster Induced by Wolbachia.

    PubMed

    Yuan, Lin-Ling; Chen, Xiulan; Zong, Qiong; Zhao, Ting; Wang, Jia-Lin; Zheng, Ya; Zhang, Ming; Wang, Zailong; Brownlie, Jeremy C; Yang, Fuquan; Wang, Yu-Feng

    2015-09-04

    To investigate the molecular mechanisms of cytoplasmic incompatibility (CI) induced by Wolbachia bacteria in Drosophila melanogaster, we applied an isobaric tags for relative and absolute quantitation (iTRAQ)-based quantitative proteomic assay to identify differentially expressed proteins extracted from spermathecae and seminal receptacles (SSR) of uninfected females mated with either 1-day-old Wolbachia-uninfected (1T) or infected males (1W) or 5-day-old infected males (5W). In total, 1317 proteins were quantified; 83 proteins were identified as having at least a 1.5-fold change in expression when 1W was compared with 1T. Differentially expressed proteins were related to metabolism, immunity, and reproduction. Wolbachia changed the expression of seminal fluid proteins (Sfps). Wolbachia may disrupt the abundance of proteins in SSR by affecting ubiquitin-proteasome-mediated proteolysis. Knocking down two Sfp genes (CG9334 and CG2668) in Wolbachia-free males resulted in significantly lower embryonic hatch rates with a phenotype of chromatin bridges. Wolbachia-infected females may rescue the hatch rates. This suggests that the changed expression of some Sfps may be one of the mechanisms of CI induced by Wolbachia. This study provides a panel of candidate proteins that may be involved in the interaction between Wolbachia and their insect hosts and, through future functional studies, may help to elucidate the underlying mechanisms of Wolbachia-induced CI.

  6. Bidirectional cytoplasmic incompatibility caused by Wolbachia in the terrestrial isopod Porcellio dilatatus.

    PubMed

    Sicard, Mathieu; Bouchon, Didier; Ceyrac, Laura; Raimond, Roland; Thierry, Magali; Le Clec'h, Winka; Marcadé, Isabelle; Caubet, Yves; Grève, Pierre

    2014-09-01

    In the terrestrial isopod species Porcellio dilatatus, unidirectional Cytoplasmic Incompatibility (CI) between two morphs (P. d. dilatatus and P. d. petiti) caused by a Wolbachia strain (wPet) infecting the morph P. d. petiti has been previously described by experiments initiated four decades ago. Here, we studied another Wolbachia that has been recently detected in a population of the morph P. d. dilatatus. The MLST markers reveal that this Wolbachia is a new strain called wDil distinct from wPet also belonging to the isopod clade of Wolbachia. Quantifications of both Wolbachia strains in the gonads of the two P. dilatatus morphs revealed that all males exhibit similar Wolbachia titers while the titers in females depend on the Wolbachia strain they host. Crossing experiments showed that both wDil and wPet induced partial unidirectional CI with different intensities. Moreover, these two strains induced bidirectional CI when individuals were both infected with one of the two different Wolbachia strains. This way, we demonstrated that P. dilatatus can be infected by two closely related Wolbachia strains (wDil and wPet), that seem to have different modification-rescue systems.

  7. Experimental evolution reveals habitat-specific fitness dynamics among Wolbachia clades in Drosophila melanogaster.

    PubMed

    Versace, Elisabetta; Nolte, Viola; Pandey, Ram Vinay; Tobler, Ray; Schlötterer, Christian

    2014-02-01

    The diversity and infection dynamics of the endosymbiont Wolbachia can be influenced by many factors, such as transmission rate, cytoplasmic incompatibility, environment, selection and genetic drift. The interplay of these factors in natural populations can result in heterogeneous infection patterns with substantial differences between populations and strains. The causes of these heterogeneities are not yet understood, partly due to the complexity of natural environments. We present experimental evolution as a new approach to study Wolbachia infection dynamics in replicate populations exposed to a controlled environment. A natural Drosophila melanogaster population infected with strains of Wolbachia belonging to different clades evolved in two laboratory environments (hot and cold) for 1.5 years. In both treatments, the rate of Wolbachia infection increased until fixation. In the hot environment, the relative frequency of different Wolbachia clades remained stable over 37 generations. In the cold environment, however, we observed marked changes in the composition of the Wolbachia population: within 15 generations, one Wolbachia clade increased more than 50% in frequency, whereas the other two clades decreased in frequency, resulting in the loss of one clade. The frequency change was highly reproducible not only among replicates, but also when flies that evolved for 42 generations in the hot environment were transferred to the cold environment. These results document how environmental factors can affect the composition of Wolbachia in D. melanogaster. The high reproducibility of the pattern suggests that experimental evolution studies can efficiently determine the functional basis of habitat-specific fitness among Wolbachia strains.

  8. Mitochondrial DNA variants help monitor the dynamics of Wolbachia invasion into host populations.

    PubMed

    Yeap, H L; Rašić, G; Endersby-Harshman, N M; Lee, S F; Arguni, E; Le Nguyen, H; Hoffmann, A A

    2016-03-01

    Wolbachia is the most widespread endosymbiotic bacterium of insects and other arthropods that can rapidly invade host populations. Deliberate releases of Wolbachia into natural populations of the dengue fever mosquito, Aedes aegypti, are used as a novel biocontrol strategy for dengue suppression. Invasion of Wolbachia through the host population relies on factors such as high fidelity of the endosymbiont transmission and limited immigration of uninfected individuals, but these factors can be difficult to measure. One way of acquiring relevant information is to consider mitochondrial DNA (mtDNA) variation alongside Wolbachia in field-caught mosquitoes. Here we used diagnostic mtDNA markers to differentiate infection-associated mtDNA haplotypes from those of the uninfected mosquitoes at release sites. Unique haplotypes associated with Wolbachia were found at locations outside Australia. We also performed mathematical and qualitative analyses including modelling the expected dynamics of the Wolbachia and mtDNA variants during and after a release. Our analyses identified key features in haplotype frequency patterns to infer the presence of imperfect maternal transmission of Wolbachia, presence of immigration and possibly incomplete cytoplasmic incompatibility. We demonstrate that ongoing screening of the mtDNA variants should provide information on maternal leakage and immigration, particularly in releases outside Australia. As we demonstrate in a case study, our models to track the Wolbachia dynamics can be successfully applied to temporal studies in natural populations or Wolbachia release programs, as long as there is co-occurring mtDNA variation that differentiates infected and uninfected populations.

  9. [Drosophila melanogaster Cell Culture as an Experimental Model to Study Recombination in Wolbachia pipientis].

    PubMed

    Goryacheva, I I; Gorelova, T V; Andrianov, B V

    2015-12-01

    Wolbachiapipientis is an obligate intracellular endosymbiont that commonly infects arthropods. Comparative genomic studies of Wolbachia reveal traces of numerous events of intergenic and intragenic recombination. The molecular mechanisms of recombination in Wolbachia are not currently known. We conducted experimental verification of the possibility of recombination of two strains of Wolbachia: wMel and wRi, after using these strains for double infection of the Dm2008Wb1 (D. melanogaster) cell culture clone permissive to Wolbachia. We obtained cell culture subclones with double Wolbachia infection and subclones infected only by strain wMel. Dual infection with the Wolbachia strains wMel and wRi has been stably maintained in the subclones for two years. Multilocus sequence typing (MLST) of the obtained subclones revealed the presence of dual infection for all five Wolbachia genes used for MLST Cloning and nucleotide sequence analysis of individual forms of the fbpA gene of Wolbachia from cell clones with dual infection showed intragenic recombination events between strains wMel and wRi, which occurred in the permanent D. melanogaster culture cell culture. The fact that putative recombination sites contain no insertions of nucleotide sequences of phages or IS elements, as well as the asymmetrical character of recombinants, favors the hypothesis that gene conversion is the most probable molecular mechanism of recombination in Wolbachia.

  10. Inter-population variation for Wolbachia induced reproductive incompatibility in the haplodiploid mite Tetranychus urticae.

    PubMed

    Suh, Eunho; Sim, Cheolho; Park, Jung-Joon; Cho, Kijong

    2015-01-01

    Recent studies have revealed diverse patterns of cytoplasmic incompatibility (CI) induced by Wolbachia in the two spotted spider mite (Tetranychus urticae Koch). The mechanism of CI consists of two steps: modification (mod) of sperm of infected males and the rescue (resc) of these chromosomes by Wolbachia in the egg, which results in female embryonic mortality (FM), male development (MD) or no CI. Our study reports that Wolbachia infections were highly prevalent infecting all T. urticae populations from various crops in 14 commercial greenhouses in Korea, with two Wolbachia strains expressing distinctive phenotypic effects on hosts. Analyses for wsp gene sequences obtained from collected mite populations revealed all sequences were categorized into two groups (group W1 and W2) discriminated by three diagnostic nucleotides while all Wolbachia strains belonged to the subgroup Ori in Wolbachia supergroup B. Host plants of each mite population were also generally correlated this grouping. Various mating experiments with two mite populations from each group showed that CI patterns and host plants of the mite populations were completely matched with the grouping; no CI (mod(-)resc(+)) for group W1 and mixed pattern of FM and MD (mod(+)resc(+)) for group W2. No distinct changes in fecundity or sex ratio due to Wolbachia infections were observed in four mite populations regardless of Wolbachia grouping. Our study suggests a potential correlation between phenotypic effect of Wolbachia infection and its genetic diversity associated with host plants in Korean mite populations.

  11. 33 CFR 104.106 - Passenger access area.

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ..., or cruise ship may designate areas within the vessel as passenger access areas. (b) A passenger... measures for access control, of a ferry, passenger vessel, or cruise ship that is open to passengers. It...

  12. 33 CFR 104.106 - Passenger access area.

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ..., or cruise ship may designate areas within the vessel as passenger access areas. (b) A passenger... measures for access control, of a ferry, passenger vessel, or cruise ship that is open to passengers. It...

  13. 33 CFR 104.106 - Passenger access area.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ..., or cruise ship may designate areas within the vessel as passenger access areas. (b) A passenger... measures for access control, of a ferry, passenger vessel, or cruise ship that is open to passengers. It...

  14. 33 CFR 104.106 - Passenger access area.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ..., or cruise ship may designate areas within the vessel as passenger access areas. (b) A passenger... measures for access control, of a ferry, passenger vessel, or cruise ship that is open to passengers. It...

  15. 33 CFR 104.106 - Passenger access area.

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ..., or cruise ship may designate areas within the vessel as passenger access areas. (b) A passenger... measures for access control, of a ferry, passenger vessel, or cruise ship that is open to passengers. It...

  16. Locating influential nodes in complex networks

    NASA Astrophysics Data System (ADS)

    Malliaros, Fragkiskos D.; Rossi, Maria-Evgenia G.; Vazirgiannis, Michalis

    2016-01-01

    Understanding and controlling spreading processes in networks is an important topic with many diverse applications, including information dissemination, disease propagation and viral marketing. It is of crucial importance to identify which entities act as influential spreaders that can propagate information to a large portion of the network, in order to ensure efficient information diffusion, optimize available resources or even control the spreading. In this work, we capitalize on the properties of the K-truss decomposition, a triangle-based extension of the core decomposition of graphs, to locate individual influential nodes. Our analysis on real networks indicates that the nodes belonging to the maximal K-truss subgraph show better spreading behavior compared to previously used importance criteria, including node degree and k-core index, leading to faster and wider epidemic spreading. We further show that nodes belonging to such dense subgraphs, dominate the small set of nodes that achieve the optimal spreading in the network.

  17. Locating influential nodes in complex networks

    PubMed Central

    Malliaros, Fragkiskos D.; Rossi, Maria-Evgenia G.; Vazirgiannis, Michalis

    2016-01-01

    Understanding and controlling spreading processes in networks is an important topic with many diverse applications, including information dissemination, disease propagation and viral marketing. It is of crucial importance to identify which entities act as influential spreaders that can propagate information to a large portion of the network, in order to ensure efficient information diffusion, optimize available resources or even control the spreading. In this work, we capitalize on the properties of the K-truss decomposition, a triangle-based extension of the core decomposition of graphs, to locate individual influential nodes. Our analysis on real networks indicates that the nodes belonging to the maximal K-truss subgraph show better spreading behavior compared to previously used importance criteria, including node degree and k-core index, leading to faster and wider epidemic spreading. We further show that nodes belonging to such dense subgraphs, dominate the small set of nodes that achieve the optimal spreading in the network. PMID:26776455

  18. 77 FR 38248 - Passenger Train Emergency Preparedness

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-06-27

    ... Federal Railroad Administration 49 CFR Part 239 Passenger Train Emergency Preparedness AGENCY: Federal... (NPRM). SUMMARY: FRA is proposing to revise its regulations for passenger train emergency preparedness... responders during emergency situations receive initial and periodic training and are subject to...

  19. Absence of Wolbachia endobacteria in the human parasitic nematode Dracunculus medinensis and two related Dracunculus species infecting wildlife

    PubMed Central

    2014-01-01

    Background Wolbachia endosymbionts are a proven target for control of human disease caused by filarial nematodes. However, little is known about the occurrence of Wolbachia in taxa closely related to the superfamily Filarioidea. Our study addressed the status of Wolbachia presence in members of the superfamily Dracunculoidea by screening the human parasite Dracunculus medinensis and related species from wildlife for Wolbachia. Findings D. medinensis, D. lutrae and D. insignis specimens were all negative for Wolbachia colonization by PCR screening for the Wolbachia ftsZ, 16S rRNA and Wolbachia surface protein (wsp) sequences. The quality and purity of the DNA preparations was confirmed by amplification of nematode 18S rRNA and cytochrome c oxidase subunit I sequences. Furthermore, Wolbachia endobacteria were not detected by whole mount fluorescence staining, or by immunohistochemistry using a Wolbachia-specific antiserum. In contrast, positive control Brugia malayi worms were shown to harbour Wolbachia by PCR, fluorescence staining and immunohistochemistry. Conclusions Three examined species of Dracunculus showed no evidence of Wolbachia endobacteria. This supports that members of the superfamily Dracunculoidea are free of Wolbachia. Within the order Spirurida, these endosymbionts appear restricted to the Filarioidea. PMID:24685011

  20. Wolbachia-mediated antiviral protection in Drosophila larvae and adults following oral infection.

    PubMed

    Stevanovic, Aleksej L; Arnold, Pieter A; Johnson, Karyn N

    2015-12-01

    Understanding viral dynamics in arthropods is of great importance when designing models to describe how viral spread can influence arthropod populations. The endosymbiotic bacterium Wolbachia spp., which is present in up to 40% of all insect species, has the ability to alter viral dynamics in both Drosophila spp. and mosquitoes, a feature that in mosquitoes may be utilized to limit spread of important arboviruses. To understand the potential effect of Wolbachia on viral dynamics in nature, it is important to consider the impact of natural routes of virus infection on Wolbachia antiviral effects. Using adult Drosophila strains, we show here that Drosophila-Wolbachia associations that have previously been shown to confer antiviral protection following systemic viral infection also confer protection against virus-induced mortality following oral exposure to Drosophila C virus in adults. Interestingly, a different pattern was observed when the same fly lines were challenged with the virus when still larvae. Analysis of the four Drosophila-Wolbachia associations that were protective in adults indicated that only the w1118-wMelPop association conferred protection in larvae following oral delivery of the virus. Analysis of Wolbachia density using quantitative PCR (qPCR) showed that a high Wolbachia density was congruent with antiviral protection in both adults and larvae. This study indicates that Wolbachia-mediated protection may vary between larval and adult stages of a given Wolbachia-host combination and that the variations in susceptibility by life stage correspond with Wolbachia density. The differences in the outcome of virus infection are likely to influence viral dynamics in Wolbachia-infected insect populations in nature and could also have important implications for the transmission of arboviruses in mosquito populations.

  1. Wolbachia-Mediated Antiviral Protection in Drosophila Larvae and Adults following Oral Infection

    PubMed Central

    Stevanovic, Aleksej L.; Arnold, Pieter A.

    2015-01-01

    Understanding viral dynamics in arthropods is of great importance when designing models to describe how viral spread can influence arthropod populations. The endosymbiotic bacterium Wolbachia spp., which is present in up to 40% of all insect species, has the ability to alter viral dynamics in both Drosophila spp. and mosquitoes, a feature that in mosquitoes may be utilized to limit spread of important arboviruses. To understand the potential effect of Wolbachia on viral dynamics in nature, it is important to consider the impact of natural routes of virus infection on Wolbachia antiviral effects. Using adult Drosophila strains, we show here that Drosophila-Wolbachia associations that have previously been shown to confer antiviral protection following systemic viral infection also confer protection against virus-induced mortality following oral exposure to Drosophila C virus in adults. Interestingly, a different pattern was observed when the same fly lines were challenged with the virus when still larvae. Analysis of the four Drosophila-Wolbachia associations that were protective in adults indicated that only the w1118-wMelPop association conferred protection in larvae following oral delivery of the virus. Analysis of Wolbachia density using quantitative PCR (qPCR) showed that a high Wolbachia density was congruent with antiviral protection in both adults and larvae. This study indicates that Wolbachia-mediated protection may vary between larval and adult stages of a given Wolbachia-host combination and that the variations in susceptibility by life stage correspond with Wolbachia density. The differences in the outcome of virus infection are likely to influence viral dynamics in Wolbachia-infected insect populations in nature and could also have important implications for the transmission of arboviruses in mosquito populations. PMID:26407882

  2. 49 CFR 523.4 - Passenger automobile.

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ... 49 Transportation 6 2010-10-01 2010-10-01 false Passenger automobile. 523.4 Section 523.4... ADMINISTRATION, DEPARTMENT OF TRANSPORTATION VEHICLE CLASSIFICATION § 523.4 Passenger automobile. A passenger automobile is any automobile (other than an automobile capable of off-highway operation)...

  3. 78 FR 38101 - Passenger Vessels Accessibility Guidelines

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-06-25

    ...We, the Architectural and Transportation Barriers Compliance Board (Access Board), are proposing accessibility guidelines for the construction and alteration of passenger vessels covered by the Americans with Disabilities Act (ADA) to ensure that the vessels are readily accessible to and usable by passengers with disabilities. The guidelines would apply to passenger vessels, other than ferries......

  4. 14 CFR 91.519 - Passenger briefing.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... Regulations require passenger compliance with lighted passenger information signs and no smoking placards, prohibit smoking in lavatories, and require compliance with crewmember instructions with regard to these... briefed on— (1) Smoking. Each passenger shall be briefed on when, where, and under what conditions...

  5. 14 CFR 91.519 - Passenger briefing.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... Regulations require passenger compliance with lighted passenger information signs and no smoking placards, prohibit smoking in lavatories, and require compliance with crewmember instructions with regard to these... briefed on— (1) Smoking. Each passenger shall be briefed on when, where, and under what conditions...

  6. 49 CFR 523.4 - Passenger automobile.

    Code of Federal Regulations, 2012 CFR

    2012-10-01

    ... 49 Transportation 6 2012-10-01 2012-10-01 false Passenger automobile. 523.4 Section 523.4... ADMINISTRATION, DEPARTMENT OF TRANSPORTATION VEHICLE CLASSIFICATION § 523.4 Passenger automobile. A passenger automobile is any automobile (other than an automobile capable of off-highway operation)...

  7. 49 CFR 523.4 - Passenger automobile.

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ... 49 Transportation 6 2013-10-01 2013-10-01 false Passenger automobile. 523.4 Section 523.4... ADMINISTRATION, DEPARTMENT OF TRANSPORTATION VEHICLE CLASSIFICATION § 523.4 Passenger automobile. A passenger automobile is any automobile (other than an automobile capable of off-highway operation)...

  8. 49 CFR 523.4 - Passenger automobile.

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    ... 49 Transportation 6 2014-10-01 2014-10-01 false Passenger automobile. 523.4 Section 523.4... ADMINISTRATION, DEPARTMENT OF TRANSPORTATION VEHICLE CLASSIFICATION § 523.4 Passenger automobile. A passenger automobile is any automobile (other than an automobile capable of off-highway operation)...

  9. 49 CFR 523.4 - Passenger automobile.

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ... 49 Transportation 6 2011-10-01 2011-10-01 false Passenger automobile. 523.4 Section 523.4... ADMINISTRATION, DEPARTMENT OF TRANSPORTATION VEHICLE CLASSIFICATION § 523.4 Passenger automobile. A passenger automobile is any automobile (other than an automobile capable of off-highway operation)...

  10. 14 CFR 125.217 - Passenger information.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... Seat Belt” sign is lighted. (d) Each passenger shall comply with instructions given him or her by... them when smoking is prohibited and when safety belts must be fastened. The signs must be so... passenger or crewmember smoke in any lavatory. (c) Each passenger required by § 125.211(b) to occupy a...

  11. 14 CFR 125.217 - Passenger information.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... Seat Belt” sign is lighted. (d) Each passenger shall comply with instructions given him or her by... them when smoking is prohibited and when safety belts must be fastened. The signs must be so... passenger or crewmember smoke in any lavatory. (c) Each passenger required by § 125.211(b) to occupy a...

  12. Finding Wolbachia in Filarial larvae and Culicidae Mosquitoes in Upper Egypt Governorate

    PubMed Central

    Dyab, Ahmed K.; Galal, Lamia A.; Mahmoud, Abeer E.; Mokhtar, Yasser

    2016-01-01

    Wolbachia is an obligatory intracellular endosymbiotic bacterium, present in over 20% of all insects altering insect reproductive capabilities and in a wide range of filarial worms which is essential for worm survival and reproduction. In Egypt, no available data were found about Wolbachia searching for it in either mosquitoes or filarial worms. Thus, we aimed to identify the possible concurrent presence of Wolbachia within different mosquitoes and filarial parasites, in Assiut Governorate, Egypt using multiplex PCR. Initially, 6 pools were detected positive for Wolbachia by single PCR. The simultaneous detection of Wolbachia and filarial parasites (Wuchereria bancrofti, Dirofilaria immitis, and Dirofilaria repens) by multiplex PCR was spotted in 5 out of 6 pools, with an overall estimated rate of infection (ERI) of 0.24%. Unexpectedly, the highest ERI (0.53%) was for Anopheles pharoensis with related Wolbachia and W. bancrofti, followed by Aedes (0.42%) and Culex (0.26%). We also observed that Wolbachia altered Culex spp. as a primary vector for W. bancrofti to be replaced by Anopheles sp. Wolbachia within filaria-infected mosquitoes in our locality gives a hope to use bacteria as a new control trend simultaneously targeting the vector and filarial parasites. PMID:27417080

  13. Effects of Wolbachia on mitochondrial DNA variation in populations of Athetis lepigone (Lepidoptera: Noctuidae) in China

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Wolbachia are endosymbiotic bacteria that infect arthropods and incompatibility among strains can affect gene flow within host insect populations, that can result in significant host mitochondrial DNA (MtD) variation. The effects of Wolbachia infection on mtDNA variation was studied in Athetis lepi...

  14. Evidence for widespread Wolbachia infection in isopod crustaceans: molecular identification and host feminization.

    PubMed Central

    Bouchon, D; Rigaud, T; Juchault, P

    1998-01-01

    Wolbachia are maternally inherited, intracellular, alpha proteobacteria that infect a wide range of arthropods. They cause three kinds of reproductive alterations in their hosts: cytoplasmic incompatibility, parthenogenesis and feminization. There have been many studies of the distribution of Wolbachia in arthropods, but very few crustacean species are known to be infected. We investigated the prevalence of Wolbachia in 85 species from five crustacean orders. Twenty-two isopod species were found to carry these bacteria. The bacteria were found mainly in terrestrial species, suggesting that Wolbachia came from a continental environment. The evolutionary relationships between these Wolbachia strains were determined by sequencing bacterial genes and by interspecific transfers. All the bacteria associated with isopods belonged to the Wolbachia B group, based on 16S rDNA sequence data. All the terrestrial isopod symbionts in this group except one formed an independent clade. The results of interspecific transfers show evidence of specialization of Wolbachia symbionts to their isopod hosts. They also suggest that host species plays a more important role than bacterial phylogeny in determining the phenotype induced by Wolbachia infection. PMID:9684374

  15. Chaos of Wolbachia Sequences Inside the Compact Fig Syconia of Ficus benjamina (Ficus: Moraceae)

    PubMed Central

    Niu, Li-Ming; Ma, Guang-Chang; Cook, James M.; Bian, Sheng-Nan; Fu, Yue-Guan; Huang, Da-Wei

    2012-01-01

    Figs and fig wasps form a peculiar closed community in which the Ficus tree provides a compact syconium (inflorescence) habitat for the lives of a complex assemblage of Chalcidoid insects. These diverse fig wasp species have intimate ecological relationships within the closed world of the fig syconia. Previous surveys of Wolbachia, maternally inherited endosymbiotic bacteria that infect vast numbers of arthropod hosts, showed that fig wasps have some of the highest known incidences of Wolbachia amongst all insects. We ask whether the evolutionary patterns of Wolbachia sequences in this closed syconium community are different from those in the outside world. In the present study, we sampled all 17 fig wasp species living on Ficus benjamina, covering 4 families, 6 subfamilies, and 8 genera of wasps. We made a thorough survey of Wolbachia infection patterns and studied evolutionary patterns in wsp (Wolbachia Surface Protein) sequences. We find evidence for high infection incidences, frequent recombination between Wolbachia strains, and considerable horizontal transfer, suggesting rapid evolution of Wolbachia sequences within the syconium community. Though the fig wasps have relatively limited contact with outside world, Wolbachia may be introduced to the syconium community via horizontal transmission by fig wasps species that have winged males and visit the syconia earlier. PMID:23145008

  16. Using Wolbachia Releases to Estimate Aedes aegypti (Diptera: Culicidae) Population Size and Survival

    PubMed Central

    Garcia, Gabriela de Azambuja; dos Santos, Lilha Maria Barbosa; Villela, Daniel Antunes Maciel; Maciel-de-Freitas, Rafael

    2016-01-01

    Mosquitoes carrying the endosymbiont bacterium Wolbachia have been deployed in field trials as a biological control intervention due to Wolbachia effects on reducing transmission of arboviruses. We performed mark, release and recapture (MRR) experiments using Wolbachia as an internal marker with daily collections with BG-Traps during the first two weeks of releases in Rio de Janeiro, Brazil. The MRR design allowed us to investigate two critical parameters that determine whether Wolbachia would successful invade a field population: the probability of daily survival (PDS) of Wolbachia-infected Aedes aegypti females, and the wild population density during releases. Released females had a PDS of 0.82 and 0.89 in the first and second weeks, respectively, immediately after releases, which is well within the range of previous estimates of survivorship of wild mosquitoes in Rio de Janeiro. Abundance estimation of wild population varied up to 10-fold higher depending on the estimation method used (634–3565 females on the average-difference model to 6365–16188 females according to Lincoln-Petersen). Wolbachia-released mosquitoes were lower than the density estimation of their wild counterparts, irrespectively of the model used. Individually screening mosquitoes for the presence of Wolbachia reduced uncertainty on abundance estimations due to fluctuation in capturing per week. A successful invasion into local population requires Ae. aegypti fitness is unaffected by Wolbachia presence, but also reliable estimates on the population size of wild mosquitoes. PMID:27479050

  17. Removing symbiotic Wolbachia bacteria specifically inhibits oogenesis in a parasitic wasp

    PubMed Central

    Dedeine, Franck; Vavre, Fabrice; Fleury, Frédéric; Loppin, Benjamin; Hochberg, Michael E.; Boulétreau, Michel

    2001-01-01

    Wolbachia are bacteria that live in the cells of various invertebrate species to which they cause a wide range of effects on physiology and reproduction. We investigated the effect of Wolbachia infection in the parasitic wasp, Asobara tabida Nees (Hymenoptera, Braconidae). In the 13 populations tested, all individuals proved to be infected by Wolbachia. The removal of Wolbachia by antibiotic treatment had a totally unexpected effect—aposymbiotic female wasps were completely incapable of producing mature oocytes and therefore could not reproduce. In contrast, oogenesis was not affected in treated Asobara citri, a closely related species that does not harbor Wolbachia. No difference between natural symbiotic and cured individuals was found for other adult traits including male fertility, locomotor activity, and size, indicating that the effect on oogenesis is highly specific. We argue that indirect effects of the treatments used in our study (antibiotic toxicity or production of toxic agents) are very unlikely to explain the sterility of females, and we present results showing a direct relationship between oocyte production and Wolbachia density in females. We conclude that Wolbachia is necessary for oogenesis in these A. tabida strains, and this association would seem to be the first example of a transition from facultative to obligatory symbiosis in arthropod–Wolbachia associations. PMID:11353833

  18. Multi-infections of feminizing Wolbachia strains in natural populations of the terrestrial isopod Armadillidium vulgare.

    PubMed

    Valette, Victorien; Bitome Essono, Paul-Yannick; Le Clec'h, Winka; Johnson, Monique; Bech, Nicolas; Grandjean, Frédéric

    2013-01-01

    Maternally inherited Wolbachia (α-Proteobacteria) are widespread parasitic reproductive manipulators. A growing number of studies have described the presence of different Wolbachia strains within a same host. To date, no naturally occurring multiple infections have been recorded in terrestrial isopods. This is true for Armadillidium vulgare which is known to harbor non simultaneously three Wolbachia strains. Traditionally, such Wolbachia are detected by PCR amplification of the wsp gene and strains are characterized by sequencing. The presence of nucleotide deletions or insertions within the wsp gene, among these three different strains, provides the opportunity to test a novel genotyping method. Herein, we designed a new primer pair able to amplify products whose lengths are specific to each Wolbachia strain so as to detect the presence of multi-infections in A. vulgare. Experimental injections of Wolbachia strains in Wolbachia-free females were used to validate the methodology. We re-investigated, using this novel method, the infection status of 40 females sampled in 2003 and previously described as mono-infected based on the classical sequencing method. Among these females, 29 were identified as bi-infected. It is the first time that naturally occurring multiple infections of Wolbachia are detected within an individual A. vulgare host. Additionally, we resampled 6 of these populations in 2010 to check the infection status of females.

  19. Northern corn rootworm (Diabrotica barberi) populations infected by at least 5 Wolbachia strains

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Wolbachia infections are present in northern corn rootworm (Diabrotica barberi) populations from east of the Mississippi River. The boundary between infected and uninfected populations is in central Illinois. DNA sequencing of Wolbachia ftsZ and wsp segments indicates that east central Illinois popu...

  20. Cannibalism and predation as paths for horizontal passage of Wolbachia between terrestrial isopods.

    PubMed

    Le Clec'h, Winka; Chevalier, Frédéric D; Genty, Lise; Bertaux, Joanne; Bouchon, Didier; Sicard, Mathieu

    2013-01-01

    The alpha-proteobacteria Wolbachia are the most widespread endosymbionts in arthropods and nematodes. Mainly maternally inherited, these so-called sex parasites have selected several strategies that increase their vertical dispersion in host populations. However, the lack of congruence between the Wolbachia and their host phylogenies suggests frequent horizontal transfers. One way that could be used for horizontal Wolbachia transfers between individuals is predation. The aim of this study was to test whether horizontal passage of Wolbachia is possible when an uninfected terrestrial isopod eats an infected one. After having eaten Armadillidium vulgare harbouring Wolbachia, the predator-recipients (the two woodlice A. vulgare and Porcellio dilatatus dilatatus) that were initially Wolbachia-free were tested positive for the presence of Wolbachia both by quantitative PCR and Fluorescence in situ Hybridization (FISH). Even if the titers were low compared to vertically infected individuals, this constitutes the first demonstration of Wolbachia occurrence in various organs of an initially uninfected host after eating an infected one.

  1. Chaos of Wolbachia sequences inside the compact fig syconia of Ficus benjamina (Ficus: moraceae).

    PubMed

    Yang, Chun-Yan; Xiao, Jin-Hua; Niu, Li-Ming; Ma, Guang-Chang; Cook, James M; Bian, Sheng-Nan; Fu, Yue-Guan; Huang, Da-Wei

    2012-01-01

    Figs and fig wasps form a peculiar closed community in which the Ficus tree provides a compact syconium (inflorescence) habitat for the lives of a complex assemblage of Chalcidoid insects. These diverse fig wasp species have intimate ecological relationships within the closed world of the fig syconia. Previous surveys of Wolbachia, maternally inherited endosymbiotic bacteria that infect vast numbers of arthropod hosts, showed that fig wasps have some of the highest known incidences of Wolbachia amongst all insects. We ask whether the evolutionary patterns of Wolbachia sequences in this closed syconium community are different from those in the outside world. In the present study, we sampled all 17 fig wasp species living on Ficus benjamina, covering 4 families, 6 subfamilies, and 8 genera of wasps. We made a thorough survey of Wolbachia infection patterns and studied evolutionary patterns in wsp (Wolbachia Surface Protein) sequences. We find evidence for high infection incidences, frequent recombination between Wolbachia strains, and considerable horizontal transfer, suggesting rapid evolution of Wolbachia sequences within the syconium community. Though the fig wasps have relatively limited contact with outside world, Wolbachia may be introduced to the syconium community via horizontal transmission by fig wasps species that have winged males and visit the syconia earlier.

  2. The potential role of Wolbachia in controlling the transmission of emerging human arboviral infections

    PubMed Central

    Kamtchum-Tatuene, Joseph; Makepeace, Benjamin L.; Benjamin, Laura; Baylis, Matthew; Solomon, Tom

    2017-01-01

    Purpose of review Wolbachia is a genus of Gram-negative intracellular bacteria that is naturally found in more than half of all arthropod species. These bacteria cannot only reduce the fitness and the reproductive capacities of arthropod vectors, but also increase their resistance to arthropod-borne viruses (arboviruses). This article reviews the evidence supporting a Wolbachia-based strategy for controlling the transmission of dengue and other arboviral infections. Recent findings Studies conducted 1 year after the field release of Wolbachia-infected mosquitoes in Australia have demonstrated the suppression of dengue virus (DENV) replication in and dissemination by mosquitoes. Recent mathematical models show that this strategy could reduce the transmission of DENV by 70%. Consequently, the WHO is encouraging countries to boost the development and implementation of Wolbachia-based prevention strategies against other arboviral infections. However, the evidence regarding the efficacy of Wolbachia to prevent the transmission of other arboviral infections is still limited to an experimental framework with conflicting results in some cases. There is a need to demonstrate the efficacy of such strategies in the field under various climatic conditions, to select the Wolbachia strain that has the best pathogen interference/spread trade-off, and to continue to build community acceptance. Summary Wolbachia represents a promising tool for controlling the transmission of arboviral infections that needs to be developed further. Long-term environmental monitoring will be necessary for timely detection of potential changes in Wolbachia/vector/virus interactions. PMID:27849636

  3. Multi-Infections of Feminizing Wolbachia Strains in Natural Populations of the Terrestrial Isopod Armadillidium Vulgare

    PubMed Central

    Valette, Victorien; Bitome Essono, Paul-Yannick; Le Clec’h, Winka; Johnson, Monique; Bech, Nicolas; Grandjean, Frédéric

    2013-01-01

    Maternally inherited Wolbachia (α-Proteobacteria) are widespread parasitic reproductive manipulators. A growing number of studies have described the presence of different Wolbachia strains within a same host. To date, no naturally occurring multiple infections have been recorded in terrestrial isopods. This is true for Armadillidium vulgare which is known to harbor non simultaneously three Wolbachia strains. Traditionally, such Wolbachia are detected by PCR amplification of the wsp gene and strains are characterized by sequencing. The presence of nucleotide deletions or insertions within the wsp gene, among these three different strains, provides the opportunity to test a novel genotyping method. Herein, we designed a new primer pair able to amplify products whose lengths are specific to each Wolbachia strain so as to detect the presence of multi-infections in A. vulgare. Experimental injections of Wolbachia strains in Wolbachia-free females were used to validate the methodology. We re-investigated, using this novel method, the infection status of 40 females sampled in 2003 and previously described as mono-infected based on the classical sequencing method. Among these females, 29 were identified as bi-infected. It is the first time that naturally occuring multiple infections of Wolbachia are detected within an individual A. vulgare host. Additionally, we resampled 6 of these populations in 2010 to check the infection status of females. PMID:24324814

  4. The wMel strain of Wolbachia Reduces Transmission of Zika virus by Aedes aegypti

    PubMed Central

    Aliota, Matthew T.; Peinado, Stephen A.; Velez, Ivan Dario; Osorio, Jorge E.

    2016-01-01

    Zika virus (ZIKV) is causing an explosive outbreak of febrile disease in the Americas. There are no effective antiviral therapies or licensed vaccines for this virus, and mosquito control strategies have not been adequate to contain the virus. A promising candidate for arbovirus control and prevention relies on the introduction of the intracellular bacterium Wolbachia into Aedes aegypti mosquitoes. This primarily has been proposed as a tool to control dengue virus (DENV) transmission; however, evidence suggests Wolbachia infections confer protection for Ae. aegypti against other arboviruses. At present, it is unknown whether or not ZIKV can infect, disseminate, and be transmitted by Wolbachia-infected Ae. aegypti. Using Ae. aegypti infected with the wMel strain of Wolbachia that are being released in Medellin, Colombia, we report that these mosquitoes have reduced vector competence for ZIKV. These results support the use of Wolbachia biocontrol as a multivalent strategy against Ae. aegypti-transmitted viruses. PMID:27364935

  5. Wolbachia infections in natural Anopheles populations affect egg laying and negatively correlate with Plasmodium development

    PubMed Central

    Shaw, W. Robert; Marcenac, Perrine; Childs, Lauren M.; Buckee, Caroline O.; Baldini, Francesco; Sawadogo, Simon P.; Dabiré, Roch K.; Diabaté, Abdoulaye; Catteruccia, Flaminia

    2016-01-01

    The maternally inherited alpha-proteobacterium Wolbachia has been proposed as a tool to block transmission of devastating mosquito-borne infectious diseases like dengue and malaria. Here we study the reproductive manipulations induced by a recently identified Wolbachia strain that stably infects natural mosquito populations of a major malaria vector, Anopheles coluzzii, in Burkina Faso. We determine that these infections significantly accelerate egg laying but do not induce cytoplasmic incompatibility or sex-ratio distortion, two parasitic reproductive phenotypes that facilitate the spread of other Wolbachia strains within insect hosts. Analysis of 221 blood-fed A. coluzzii females collected from houses shows a negative correlation between the presence of Plasmodium parasites and Wolbachia infection. A mathematical model incorporating these results predicts that infection with these endosymbionts may reduce malaria prevalence in human populations. These data suggest that Wolbachia may be an important player in malaria transmission dynamics in Sub-Saharan Africa. PMID:27243367

  6. Wolbachia Infection Dynamics in Tribolium confusum (Coleoptera: Tenebrionidae) and Their Effects on Host Mating Behavior and Reproduction.

    PubMed

    Ming, Qing-Lei; Shen, Jia-Fei; Cheng, Chao; Liu, Chan-Min; Feng, Zhao-Jun

    2015-06-01

    Wolbachia interact with their hosts in a broad variety of relationships that range from parasitism to mutualism. To improve the understanding of complex relationships between Wolbachia and host, we performed not only mating and crossing experiments to investigate effects of Wolbachia on mate choice, mating performance, and reproduction in the confused flour beetles Tribolium confusum (Jacquelin du Val), but also quantitative PCR to determine Wolbachia spatiotemporal infection density dynamics within beetles. Wolbachia induced strong cytoplasmic incompatibility, but had no effects on male mate choice and mating performance. Compared with Wolbachia-uninfected females, infected females had very high fecundity irrespective of male's infection status. Wolbachia infection densities in beetles were higher in eggs and adults and in the reproductive tissues and abdomens, whereas Wolbachia density in adults did not differ between sexes and among different ages. These results suggest that Wolbachia have evolved mutualistic interactions with T. confusum, which provides the first evidence of Wolbachia mutualisms in this beetle species. We discussed these findings and their evolutionary implications in light of Wolbachia-host interactions.

  7. The Mutualistic Side of Wolbachia–Isopod Interactions: Wolbachia Mediated Protection Against Pathogenic Intracellular Bacteria

    PubMed Central

    Braquart-Varnier, Christine; Altinli, Mine; Pigeault, Romain; Chevalier, Frédéric D.; Grève, Pierre; Bouchon, Didier; Sicard, Mathieu

    2015-01-01

    Wolbachia is a vertically transmitted endosymbiont whose radiative success is mainly related to various host reproductive manipulations that led to consider this symbiont as a conflictual reproductive parasite. However, lately, some Wolbachia have been shown to act as beneficial symbionts by protecting hosts against a broad range of parasites. Still, this protection has been mostly demonstrated in artificial Wolbachia-host associations between partners that did not co-evolved together. Here, we tested in two terrestrial isopod species Armadillidium vulgare and Porcellio dilatatus whether resident Wolbachia (native or non-native) could confer protection during infections with Listeria ivanovii and Salmonella typhimurium and also during a transinfection with a Wolbachia strain that kills the recipient host (i.e., wVulC in P. dilatatus). Survival analyses showed that (i) A. vulgare lines hosting their native Wolbachia (wVulC) always exhibited higher survival than asymbiotic ones when infected with pathogenic bacteria (ii) P. dilatatus lines hosting their native wDil Wolbachia strain survived the S. typhimurium infection better, while lines hosting non-native wCon Wolbachia strain survived the L. ivanovii and also the transinfection with wVulC from A. vulgare better. By studying L. ivanovii and S. typhimurium loads in the hemolymph of the different host-Wolbachia systems, we showed that (i) the difference in survival between lines after L. ivanovii infections were not linked to the difference between their pathogenic bacterial loads, and (ii) the difference in survival after S. typhimurium infections corresponds to lower loads of pathogenic bacteria. Overall, our results demonstrate a beneficial effect of Wolbachia on survival of terrestrial isopods when infected with pathogenic intracellular bacteria. This protective effect may rely on different mechanisms depending on the resident symbiont and the invasive bacteria interacting together within the hosts. PMID:26733946

  8. Glucose and Glycogen Metabolism in Brugia malayi Is Associated with Wolbachia Symbiont Fitness

    PubMed Central

    Voronin, Denis; Bachu, Saheed; Shlossman, Michael; Unnasch, Thomas R.; Ghedin, Elodie; Lustigman, Sara

    2016-01-01

    Wolbachia are endosymbiotic bacteria found in the majority of arthropods and filarial nematodes of medical and veterinary importance. They have evolved a wide range of symbiotic associations. In filarial nematodes that cause human lymphatic filariasis (Wuchereria bancrofti, Brugia malayi) or onchocerciasis (Onchocerca volvulus), Wolbachia are important for parasite development, reproduction and survival. The symbiotic bacteria rely in part on nutrients and energy sources provided by the host. Genomic analyses suggest that the strain of Wolbachia found in B. malayi (wBm) lacks the genes for two glycolytic enzymes—6-phosphofructokinase and pyruvate kinase—and is thus potentially unable to convert glucose into pyruvate, an important substrate for energy generation. The Wolbachia surface protein, wBm00432, is complexed to six B. malayi glycolytic enzymes, including aldolase. In this study we characterized two B. malayi aldolase isozymes and found that their expression is dependent on Wolbachia fitness and number. We confirmed by immuno-transmission electron microscopy that aldolase is associated with the Wolbachia surface. RNAi experiments suggested that aldolase-2 plays a significant role in both Wolbachia survival and embryogenesis in B. malayi. Treatment with doxycycline reduced Wolbachia fitness and increased the amount of both glucose and glycogen detected in the filarial parasite, indicating that glucose metabolism and glycogen storage in B. malayi are associated with Wolbachia fitness. This metabolic co-dependency between Wolbachia and its filarial nematode indicates that glycolysis could be a shared metabolic pathway between the bacteria and B. malayi, and thus a potential new target for anti-filarial therapy. PMID:27078260

  9. Onchocerca armillata contains the endosymbiotic bacterium Wolbachia and elicits a limited inflammatory response

    PubMed Central

    Neary, Joseph M.; Trees, Alexander J.; Ekale, David D.; Tanya, Vincent N.; Hetzel, Udo; Makepeace, Benjamin L.

    2010-01-01

    Human onchocerciasis, also known as River Blindness, is a debilitating disease caused by the filarial nematode Onchocerca volvulus. Many, but not all, filarial nematodes carry within their tissues endosymbiotic, Rickettsia-like bacteria of the genus Wolbachia. Onchocerca spp. infections in cattle offer the most relevant, analogous host–parasite model system. West African cattle are commonly co-infected with four Onchocerca spp.; two of these are Wolbachia-positive (Onchocerca gutturosa and Onchocerca ochengi), and the remainder are of unknown Wolbachia status (Onchocerca dukei and Onchocerca armillata). Previous studies have suggested that worm survival is dependent on this bacterium. O. armillata, an abundant parasite of African cattle that has received little attention, is a primitive species that may lack Wolbachia. The objectives of this study were to determine if O. armillata carries Wolbachia and to provide preliminary descriptions of the host inflammatory cell environment around the adult worms. The findings may support or refute the hypothesis that a prime contribution of Wolbachia is to permit long-term survival and reproduction of certain Onchocerca spp. (including O. volvulus in humans). O. armillata adult worms were found in the aorta of 90.7% of cattle (n = 54) slaughtered at an abattoir in Ngaoundéré, Adamawa Region, Cameroon. The presence of Wolbachia in O. armillata was confirmed by a specific anti-Wolbachia surface protein antibody detected using a peroxidase conjugate (immunohistochemistry) and PCR for detection of Wolbachia-specific sequences within DNA extracts from frozen worms. Tissue sections stained with haematoxylin and eosin showed the host cell response to be dominated by macrophages and fibroblasts. This is unusual compared with nodule-dwelling Wolbachia-positive Onchocerca spp., where the host response is typically characterised by granulocytes, and suggests that the mechanisms for worm survival employed by this species (which is

  10. Native Wolbachia from Aedes albopictus Blocks Chikungunya Virus Infection In Cellulo.

    PubMed

    Raquin, Vincent; Valiente Moro, Claire; Saucereau, Yoann; Tran, Florence-Hélène; Potier, Patrick; Mavingui, Patrick

    2015-01-01

    Wolbachia, a widespread endosymbiont of terrestrial arthropods, can protect its host against viral and parasitic infections, a phenotype called "pathogen blocking". However, in some cases Wolbachia may have no effect or even enhance pathogen infection, depending on the host-Wolbachia-pathogen combination. The tiger mosquito Aedes albopictus is naturally infected by two strains of Wolbachia, wAlbA and wAlbB, and is a competent vector for different arboviruses such as dengue virus (DENV) and chikungunya virus (CHIKV). Interestingly, it was shown in some cases that Ae. albopictus native Wolbachia strains are able to inhibit DENV transmission by limiting viral replication in salivary glands, but no such impact was measured on CHIKV replication in vivo. To better understand the Wolbachia/CHIKV/Ae. albopictus interaction, we generated a cellular model using Ae. albopictus derived C6/36 cells that we infected with the wAlbB strain. Our results indicate that CHIKV infection is negatively impacted at both RNA replication and virus assembly/secretion steps in presence of wAlbB. Using FISH, we observed CHIKV and wAlbB in the same mosquito cells, indicating that the virus is still able to enter the cell in the presence of the bacterium. Further work is needed to decipher molecular pathways involved in Wolbachia-CHIKV interaction at the cellular level, but this cellular model can be a useful tool to study the mechanism behind virus blocking phenotype induced by Wolbachia. More broadly, this put into question the ecological role of Wolbachia symbiont in Ae. albopictus, but also the ability of the CHIKV to counteract Wolbachia's antiviral potential in vivo.

  11. The Endosymbiotic Bacterium Wolbachia Selectively Kills Male Hosts by Targeting the Masculinizing Gene.

    PubMed

    Fukui, Takahiro; Kawamoto, Munetaka; Shoji, Keisuke; Kiuchi, Takashi; Sugano, Sumio; Shimada, Toru; Suzuki, Yutaka; Katsuma, Susumu

    2015-07-01

    Pathogens are known to manipulate the reproduction and development of their hosts for their own benefit. Wolbachia is an endosymbiotic bacterium that infects a wide range of insect species. Wolbachia is known as an example of a parasite that manipulates the sex of its host's progeny. Infection of Ostrinia moths by Wolbachia causes the production of all-female progeny, however, the mechanism of how Wolbachia accomplishes this male-specific killing is unknown. Here we show for the first time that Wolbachia targets the host masculinizing gene of Ostrinia to accomplish male-killing. We found that Wolbachia-infected O. furnacalis embryos do not express the male-specific splice variant of doublesex, a gene which acts at the downstream end of the sex differentiation cascade, throughout embryonic development. Transcriptome analysis revealed that Wolbachia infection markedly reduces the mRNA level of Masc, a gene that encodes a protein required for both masculinization and dosage compensation in the silkworm Bombyx mori. Detailed bioinformatic analysis also elucidated that dosage compensation of Z-linked genes fails in Wolbachia-infected O. furnacalis embryos, a phenomenon that is extremely similar to that observed in Masc mRNA-depleted male embryos of B. mori. Finally, injection of in vitro transcribed Masc cRNA into Wolbachia-infected embryos rescued male progeny. Our results show that Wolbachia-induced male-killing is caused by a failure of dosage compensation via repression of the host masculinizing gene. Our study also shows a novel strategy by which a pathogen hijacks the host sex determination cascade.

  12. Host tissues as microhabitats for Wolbachia and quantitative insights into the bacterial community in terrestrial isopods.

    PubMed

    Dittmer, J; Beltran-Bech, S; Lesobre, J; Raimond, M; Johnson, M; Bouchon, D

    2014-05-01

    Animal-bacterial symbioses are highly dynamic in terms of multipartite interactions, both between the host and its symbionts as well as between the different bacteria constituting the symbiotic community. These interactions will be reflected by the titres of the individual bacterial taxa, for example via host regulation of bacterial loads or competition for resources between symbionts. Moreover, different host tissues represent heterogeneous microhabitats for bacteria, meaning that host-associated bacteria might establish tissue-specific bacterial communities. Wolbachia are widespread endosymbiotic bacteria, infecting a large number of arthropods and filarial nematodes. However, relatively little is known regarding direct interactions between Wolbachia and other bacteria. This study represents the first quantitative investigation of tissue-specific Wolbachia-microbiota interactions in the terrestrial isopod Armadillidium vulgare. To this end, we obtained a more complete picture of the Wolbachia distribution patterns across all major host tissues, integrating all three feminizing Wolbachia strains (wVulM, wVulC, wVulP) identified to date in this host. Interestingly, the different Wolbachia strains exhibited strain-specific tissue distribution patterns, with wVulM reaching lower titres in most tissues. These patterns were consistent across different host genetic backgrounds and might reflect different co-evolutionary histories between the Wolbachia strains and A. vulgare. Moreover, Wolbachia-infected females carried higher total bacterial loads in several, but not all, tissues, irrespective of the Wolbachia strain. Taken together, this quantitative approach indicates that Wolbachia is part of a potentially more diverse bacterial community, as exemplified by the presence of highly abundant bacterial taxa in the midgut caeca of several A. vulgare populations.

  13. Glucose and Glycogen Metabolism in Brugia malayi Is Associated with Wolbachia Symbiont Fitness.

    PubMed

    Voronin, Denis; Bachu, Saheed; Shlossman, Michael; Unnasch, Thomas R; Ghedin, Elodie; Lustigman, Sara

    2016-01-01

    Wolbachia are endosymbiotic bacteria found in the majority of arthropods and filarial nematodes of medical and veterinary importance. They have evolved a wide range of symbiotic associations. In filarial nematodes that cause human lymphatic filariasis (Wuchereria bancrofti, Brugia malayi) or onchocerciasis (Onchocerca volvulus), Wolbachia are important for parasite development, reproduction and survival. The symbiotic bacteria rely in part on nutrients and energy sources provided by the host. Genomic analyses suggest that the strain of Wolbachia found in B. malayi (wBm) lacks the genes for two glycolytic enzymes--6-phosphofructokinase and pyruvate kinase--and is thus potentially unable to convert glucose into pyruvate, an important substrate for energy generation. The Wolbachia surface protein, wBm00432, is complexed to six B. malayi glycolytic enzymes, including aldolase. In this study we characterized two B. malayi aldolase isozymes and found that their expression is dependent on Wolbachia fitness and number. We confirmed by immuno-transmission electron microscopy that aldolase is associated with the Wolbachia surface. RNAi experiments suggested that aldolase-2 plays a significant role in both Wolbachia survival and embryogenesis in B. malayi. Treatment with doxycycline reduced Wolbachia fitness and increased the amount of both glucose and glycogen detected in the filarial parasite, indicating that glucose metabolism and glycogen storage in B. malayi are associated with Wolbachia fitness. This metabolic co-dependency between Wolbachia and its filarial nematode indicates that glycolysis could be a shared metabolic pathway between the bacteria and B. malayi, and thus a potential new target for anti-filarial therapy.

  14. Regulation of arginine methyltransferase 3 by a Wolbachia-induced microRNA in Aedes aegypti and its effect on Wolbachia and dengue virus replication.

    PubMed

    Zhang, Guangmei; Hussain, Mazhar; Asgari, Sassan

    2014-10-01

    The gram-negative endosymbiotic bacteria, Wolbachia, have been found to colonize a wide range of invertebrates, including over 40% of insect species. Best known for host reproductive manipulations, some strains of Wolbachia have been shown to reduce the host life span by about 50% and inhibit replication and transmission of dengue virus (DENV) in the mosquito vector, Aedes aegypti. The molecular mechanisms underlying these effects still are not well understood. Our previous studies showed that Wolbachia uses host microRNAs (miRNAs) to manipulate host gene expression for its efficient maintenance and limiting replication of DENV in Ae. aegypti. Protein arginine methyltransferases are structurally and functionally conserved proteins from yeast to human. In mammals, it has been reported that protein arginine methyltransferases such as PRMT1, 5 and 6 could regulate replication of different viruses. Ae. aegypti contains eight members of protein arginine methyltransferases (AaArgM1-8). Here, we show that the wMelPop strain of Wolbachia introduced into Ae. aegypti significantly induces the expression of AaArgM3. Interestingly, we found that Wolbachia uses aae-miR-2940, which is highly upregulated in Wolbachia-infected mosquitoes, to upregulate the expression of AaArgM3. Silencing of AaArgM3 in a mosquito cell line led to a significant reduction in Wolbachia replication, but had no effect on the replication of DENV. These results provide further evidence that Wolbachia uses the host miRNAs to manipulate host gene expression and facilitate colonization in Ae. aegypti mosquito.

  15. Wolbachia in the Flesh: Symbiont Intensities in Germ-Line and Somatic Tissues Challenge the Conventional View of Wolbachia Transmission Routes

    PubMed Central

    Frost, Crystal L.; Pollock, Steven W.; Smith, Judith E.; Hughes, William O. H.

    2014-01-01

    Symbionts can substantially affect the evolution and ecology of their hosts. The investigation of the tissue-specific distribution of symbionts (tissue tropism) can provide important insight into host-symbiont interactions. Among other things, it can help to discern the importance of specific transmission routes and potential phenotypic effects. The intracellular bacterial symbiont Wolbachia has been described as the greatest ever panzootic, due to the wide array of arthropods that it infects. Being primarily vertically transmitted, it is expected that the transmission of Wolbachia would be enhanced by focusing infection in the reproductive tissues. In social insect hosts, this tropism would logically extend to reproductive rather than sterile castes, since the latter constitute a dead-end for vertically transmission. Here, we show that Wolbachia are not focused on reproductive tissues of eusocial insects, and that non-reproductive tissues of queens and workers of the ant Acromyrmex echinatior, harbour substantial infections. In particular, the comparatively high intensities of Wolbachia in the haemolymph, fat body, and faeces, suggest potential for horizontal transmission via parasitoids and the faecal-oral route, or a role for Wolbachia modulating the immune response of this host. It may be that somatic tissues and castes are not the evolutionary dead-end for Wolbachia that is commonly thought. PMID:24988478

  16. Wolbachia Density and Cytoplasmic Incompatibility in Aedes albopictus: Concerns with Using Artificial Wolbachia Infection as a Vector Suppression Tool

    PubMed Central

    Calvitti, Maurizio; Marini, Francesca; Desiderio, Angiola; Puggioli, Arianna; Moretti, Riccardo

    2015-01-01

    The mosquito Aedes albopictusi is a competent vector of harmful human pathogens, including viruses causing dengue and chikungunya. Cytoplasmic incompatibility (CI) induced by endosymbiotic Wolbachia can be used to produce functionally sterile males that can be released in the field as a suppression tool against this mosquito. Because the available sexing methods are not efficient enough to avoid unintentional release of a few transinfected females, we assessed the CI pattern in crosses between wPip Wolbachia-transinfected (ARwP) females and wild-type males of Ae. albopictus in this study. Quantitative polymerase chain reaction was used to monitor the titer of the Wolbachia strains that naturally infect Ae. albopictus, that is, wAlbA and wAlbB, in age-controlled males and females. Data were coupled with incompatibility level detected when the above-mentioned males were crossed with ARwP females. Wolbachia infection titer was also monitored in samples of wild caught males. Incompatibility level was positively correlated only with wAlbA density. Crosses between wild-type males having very low wAlbA density (<0.001 wAlbA/actin copy numbers) and ARwP females were partially fertile (CIcorr = 68.06 ± 6.20). Individuals with low wAlbA titer were frequently found among sampled wild males (30%–50% depending on the site and period). ARwP males can be as considered as a very promising tool for suppressing Ae. albopictus. However, crosses between wild males having low wAlbA density and ARwP females may be partially fertile. In the case of local establishment of the transinfected mosquito line, this occurrence may favor the replacement of the wild-type mosquitoes with the ARwP line, thus reducing the long-term efficacy of incompatible insect technique. Various alternative strategies have been discussed to prevent this risk and to exploit Wolbachia as a tool to control Ae. albopictus. PMID:25812130

  17. Wolbachia density and cytoplasmic incompatibility in Aedes albopictus: concerns with using artificial Wolbachia infection as a vector suppression tool.

    PubMed

    Calvitti, Maurizio; Marini, Francesca; Desiderio, Angiola; Puggioli, Arianna; Moretti, Riccardo

    2015-01-01

    The mosquito Aedes albopictusi is a competent vector of harmful human pathogens, including viruses causing dengue and chikungunya. Cytoplasmic incompatibility (CI) induced by endosymbiotic Wolbachia can be used to produce functionally sterile males that can be released in the field as a suppression tool against this mosquito. Because the available sexing methods are not efficient enough to avoid unintentional release of a few transinfected females, we assessed the CI pattern in crosses between wPip Wolbachia-transinfected (ARwP) females and wild-type males of Ae. albopictus in this study. Quantitative polymerase chain reaction was used to monitor the titer of the Wolbachia strains that naturally infect Ae. albopictus, that is, wAlbA and wAlbB, in age-controlled males and females. Data were coupled with incompatibility level detected when the above-mentioned males were crossed with ARwP females. Wolbachia infection titer was also monitored in samples of wild caught males. Incompatibility level was positively correlated only with wAlbA density. Crosses between wild-type males having very low wAlbA density (<0.001 wAlbA/actin copy numbers) and ARwP females were partially fertile (CIcorr = 68.06 ± 6.20). Individuals with low wAlbA titer were frequently found among sampled wild males (30%-50% depending on the site and period). ARwP males can be as considered as a very promising tool for suppressing Ae. albopictus. However, crosses between wild males having low wAlbA density and ARwP females may be partially fertile. In the case of local establishment of the transinfected mosquito line, this occurrence may favor the replacement of the wild-type mosquitoes with the ARwP line, thus reducing the long-term efficacy of incompatible insect technique. Various alternative strategies have been discussed to prevent this risk and to exploit Wolbachia as a tool to control Ae. albopictus.

  18. Wolbachia-induced paternal defect in Drosophila is likely by interaction with the juvenile hormone pathway.

    PubMed

    Liu, Chen; Wang, Jia-Lin; Zheng, Ya; Xiong, En-Juan; Li, Jing-Jing; Yuan, Lin-Ling; Yu, Xiao-Qiang; Wang, Yu-Feng

    2014-06-01

    Wolbachia are endosymbionts that infect many insect species. They can manipulate the host's reproduction to increase their own maternal transmission. Cytoplasmic incompatibility (CI) is one such manipulation, which is expressed as embryonic lethality when Wolbachia-infected males mate with uninfected females. However, matings between males and females carrying the same Wolbachia strain result in viable progeny. The molecular mechanisms of CI are currently not clear. We have previously reported that the gene Juvenile hormone-inducible protein 26 (JhI-26) exhibited the highest upregulation in the 3rd instar larval testes of Drosophila melanogaster when infected by Wolbachia. This is reminiscent of an interaction between Wolbachia and juvenile hormone (JH) pathway in flies. Considering that Jhamt gene encodes JH acid methyltransferase, a key regulatory enzyme of JH biosynthesis, and that methoprene-tolerant (Met) has been regarded as the best JH receptor candidate, we first compared the expression of Jhamt and Met between Wolbachia-infected and uninfected fly testes to investigate whether Wolbachia infection influence the JH signaling pathway. We found that the expressions of Jhamt and Met were significantly increased in the presence of Wolbachia, suggesting an interaction of Wolbachia with the JH signaling pathway. Then, we found that overexpression of JhI-26 in Wolbachia-free transgenic male flies caused paternal-effect lethality that mimics the defects associated with CI. JhI-26 overexpressing males resulted in significantly decrease in hatch rate. Surprisingly, Wolbachia-infected females could rescue the egg hatch. In addition, we showed that overexpression of JhI-26 caused upregulation of the male accessory gland protein (Acp) gene CG10433, but not vice versa. This result suggests that JhI-26 may function at the upstream of CG10433. Likewise, overexpression of CG10433 also resulted in paternal-effect lethality. Both JhI-26 and CG10433 overexpressing males

  19. Model of aircraft passenger acceptance

    NASA Technical Reports Server (NTRS)

    Jacobson, I. D.

    1978-01-01

    A technique developed to evaluate the passenger response to a transportation system environment is described. Reactions to motion, noise, temperature, seating, ventilation, sudden jolts and descents are modeled. Statistics are presented for the age, sex, occupation, and income distributions of the candidates analyzed. Values are noted for the relative importance of system variables such as time savings, on-time arrival, convenience, comfort, safety, the ability to read and write, and onboard services.

  20. Wolbachia strains in cryptic species of the Anastrepha fraterculus complex (Diptera, Tephritidae) along the Neotropical Region.

    PubMed

    Prezotto, Leandro F; Perondini, André L P; Hernández-Ortiz, Vicente; Marino, Celso L; Selivon, Denise

    2017-01-01

    Infection by Wolbachia was described previously in eleven species of Anastrepha fruit flies some of which are important pests of fruticulture. One such species is the nominal Anastrepha fraterculus, the South American fruit fly, which actually comprises a complex of cryptic species. The suggestions of using Wolbachia for the control of these pest species, make imperative a more precise characterization of the existing strains of the bacteria. In this study, population samples of the A. fraterculus complex from Brazil, Argentina, Peru, Ecuador, Colombia, Guatemala and Mexico were analyzed for Wolbachia infection. The bacteria were genotyped by the MLST and WSP Typing methodologies. All samples were infected with Wolbachia of supergroup "A". For each of the five MLST genes, unique as well as already known alleles were detected. Nineteen sequence types for the concatenated sequences of the five MLST genes, and twenty wsp alleles were found in the samples. Host-specific haplotypes, shared strains among distinct hosts, and more than one strain of Wolbachia were found in some population samples. Recombination among the MLST genes and intragenic recombination between wsp haplotypes was rare. Phylogenetic analysis showed a great similarity among the Wolbachia strains in the A. fraterculus complex. However, some strains of Wolbachia are found throughout the Neotropical Region and there are specific strains in determined geographical areas.

  1. Comparative Analysis of Wolbachia Genomes Reveals Streamlining and Divergence of Minimalist Two-Component Systems

    PubMed Central

    Christensen, Steen; Serbus, Laura Renee

    2015-01-01

    Two-component regulatory systems are commonly used by bacteria to coordinate intracellular responses with environmental cues. These systems are composed of functional protein pairs consisting of a sensor histidine kinase and cognate response regulator. In contrast to the well-studied Caulobacter crescentus system, which carries dozens of these pairs, the streamlined bacterial endosymbiont Wolbachia pipientis encodes only two pairs: CckA/CtrA and PleC/PleD. Here, we used bioinformatic tools to compare characterized two-component system relays from C. crescentus, the related Anaplasmataceae species Anaplasma phagocytophilum and Ehrlichia chaffeensis, and 12 sequenced Wolbachia strains. We found the core protein pairs and a subset of interacting partners to be highly conserved within Wolbachia and these other Anaplasmataceae. Genes involved in two-component signaling were positioned differently within the various Wolbachia genomes, whereas the local context of each gene was conserved. Unlike Anaplasma and Ehrlichia, Wolbachia two-component genes were more consistently found clustered with metabolic genes. The domain architecture and key functional residues standard for two-component system proteins were well-conserved in Wolbachia, although residues that specify cognate pairing diverged substantially from other Anaplasmataceae. These findings indicate that Wolbachia two-component signaling pairs share considerable functional overlap with other α-proteobacterial systems, whereas their divergence suggests the potential for regulatory differences and cross-talk. PMID:25809075

  2. Molecular characterization of Wolbachia infection in bed bugs (Cimex lectularius) collected from several localities in France

    PubMed Central

    Akhoundi, Mohammad; Cannet, Arnaud; Loubatier, Céline; Berenger, Jean-Michel; Izri, Arezki; Marty, Pierre; Delaunay, Pascal

    2016-01-01

    Wolbachia symbionts are maternally inherited intracellular bacteria that have been detected in numerous insects including bed bugs. The objective of this study, the first epidemiological study in Europe, was to screen Wolbachia infection among Cimex lectularius collected in the field, using PCR targeting the surface protein gene (wsp), and to compare obtained Wolbachia strains with those reported from laboratory colonies of C. lectularius as well as other Wolbachia groups. For this purpose, 284 bed bug specimens were caught and studied from eight different regions of France including the suburbs of Paris, Bouches-du-Rhône, Lot-et-Garonne, and five localities in Alpes-Maritimes. Among the samples, 166 were adults and the remaining 118 were considered nymphs. In all, 47 out of 118 nymphs (40%) and 61 out of 166 adults (37%) were found positive on wsp screening. Among the positive cases, 10 samples were selected randomly for sequencing. The sequences had 100% homology with wsp sequences belonging to the F-supergroup strains of Wolbachia. Therefore, we confirm the similarity of Wolbachia strains detected in this epidemiological study to Wolbachia spp. reported from laboratory colonies of C. lectularius. PMID:27492563

  3. Molecular evidence for single Wolbachia infections among geographic strains of the flour beetle Tribolium confusum.

    PubMed Central

    Fialho, R F; Stevens, L

    1997-01-01

    Infections with the rickettsial microorganism Wolbachia are cytoplasmically inherited and occur in a wide range of insect species and several other arthropods. Wolbachia infection often results in unidirectional cytoplasmic incompatibility (CI): crosses between infected males and uninfected females are incompatible and show a reduction of progeny or complete inviability. Unidirectional CI can also occur when males harbouring two incompatible Wolbachia strains are crossed with females infected with only one of the two strains. In the flour beetle Tribolium confusum, Wolbachia infections are of particular interest because of the severity of incompatibility. Typically, no progeny results from the incompatible cross, whereas only partial incompatibility is observed in most other hosts. Werren et al. (1995a) reported that Wolbachia infections in T. confusum consist of two bacterial strains belonging to distinct phylogenic groups, based on PCR amplification and sequence analysis of the bacterial cell division gene ftsZ. However, Fialho & Stevens (1996) showed that eight strains of T. confusum were infected with a single and common incompatibility type. Here we report analysis of the ftsZ gene by specific PCR amplification. Diagnostic restriction enzyme assays revealed no evidence of double infections in 11 geographic strains of T. confusum, including the strain examined by Werren et al. (1995a). Further, sequence analysis of the Wolbachia ftsZ gene and an internal transcribed spacer (ITS) region in two of these strains displayed no nucleotide variation or evidence of polymorphisms. Results suggest that T. confusum is infected with B-group Wolbachia only. PMID:9263471

  4. Presence of extensive Wolbachia symbiont insertions discovered in the genome of its host Glossina morsitans morsitans.

    PubMed

    Brelsfoard, Corey; Tsiamis, George; Falchetto, Marco; Gomulski, Ludvik M; Telleria, Erich; Alam, Uzma; Doudoumis, Vangelis; Scolari, Francesca; Benoit, Joshua B; Swain, Martin; Takac, Peter; Malacrida, Anna R; Bourtzis, Kostas; Aksoy, Serap

    2014-04-01

    Tsetse flies (Glossina spp.) are the cyclical vectors of Trypanosoma spp., which are unicellular parasites responsible for multiple diseases, including nagana in livestock and sleeping sickness in humans in Africa. Glossina species, including Glossina morsitans morsitans (Gmm), for which the Whole Genome Sequence (WGS) is now available, have established symbiotic associations with three endosymbionts: Wigglesworthia glossinidia, Sodalis glossinidius and Wolbachia pipientis (Wolbachia). The presence of Wolbachia in both natural and laboratory populations of Glossina species, including the presence of horizontal gene transfer (HGT) events in a laboratory colony of Gmm, has already been shown. We herein report on the draft genome sequence of the cytoplasmic Wolbachia endosymbiont (cytWol) associated with Gmm. By in silico and molecular and cytogenetic analysis, we discovered and validated the presence of multiple insertions of Wolbachia (chrWol) in the host Gmm genome. We identified at least two large insertions of chrWol, 527,507 and 484,123 bp in size, from Gmm WGS data. Southern hybridizations confirmed the presence of Wolbachia insertions in Gmm genome, and FISH revealed multiple insertions located on the two sex chromosomes (X and Y), as well as on the supernumerary B-chromosomes. We compare the chrWol insertions to the cytWol draft genome in an attempt to clarify the evolutionary history of the HGT events. We discuss our findings in light of the evolution of Wolbachia infections in the tsetse fly and their potential impacts on the control of tsetse populations and trypanosomiasis.

  5. Longicorn beetle that vectors pinewood nematode carries many Wolbachia genes on an autosome

    PubMed Central

    Aikawa, Takuya; Anbutsu, Hisashi; Nikoh, Naruo; Kikuchi, Taisei; Shibata, Fukashi; Fukatsu, Takema

    2009-01-01

    Monochamus alternatus is the longicorn beetle notorious as a vector of the pinewood nematode that causes the pine wilt disease. When two populations of M. alternatus were subjected to diagnostic polymerase chain reaction (PCR) detection of four Wolbachia genes, only the ftsZ gene was detected from one of the populations. The Wolbachia ftsZ gene persisted even after larvae were fed with a tetracycline-containing diet for six weeks. The inheritance of the ftsZ gene was not maternal but biparental, exhibiting a typical Mendelian pattern. The ftsZ gene titres in homozygotic ftsZ+ insects were nearly twice as high as those in heterozygotic ftsZ+ insects. Exhaustive PCR surveys revealed that 31 and 30 of 214 Wolbachia genes examined were detected from the two insect populations, respectively. Many of these Wolbachia genes contained stop codon(s) and/or frame shift(s). Fluorescent in situ hybridization confirmed the location of the Wolbachia genes on an autosome. On the basis of these results, we conclude that a large Wolbachia genomic region has been transferred to and located on an autosome of M. alternatus. The discovery of massive gene transfer from Wolbachia to M. alternatus would provide further insights into the evolution and fate of laterally transferred endosymbiont genes in multicellular host organisms. PMID:19692404

  6. Longicorn beetle that vectors pinewood nematode carries many Wolbachia genes on an autosome.

    PubMed

    Aikawa, Takuya; Anbutsu, Hisashi; Nikoh, Naruo; Kikuchi, Taisei; Shibata, Fukashi; Fukatsu, Takema

    2009-11-07

    Monochamus alternatus is the longicorn beetle notorious as a vector of the pinewood nematode that causes the pine wilt disease. When two populations of M. alternatus were subjected to diagnostic polymerase chain reaction (PCR) detection of four Wolbachia genes, only the ftsZ gene was detected from one of the populations. The Wolbachia ftsZ gene persisted even after larvae were fed with a tetracycline-containing diet for six weeks. The inheritance of the ftsZ gene was not maternal but biparental, exhibiting a typical Mendelian pattern. The ftsZ gene titres in homozygotic ftsZ(+) insects were nearly twice as high as those in heterozygotic ftsZ(+) insects. Exhaustive PCR surveys revealed that 31 and 30 of 214 Wolbachia genes examined were detected from the two insect populations, respectively. Many of these Wolbachia genes contained stop codon(s) and/or frame shift(s). Fluorescent in situ hybridization confirmed the location of the Wolbachia genes on an autosome. On the basis of these results, we conclude that a large Wolbachia genomic region has been transferred to and located on an autosome of M. alternatus. The discovery of massive gene transfer from Wolbachia to M. alternatus would provide further insights into the evolution and fate of laterally transferred endosymbiont genes in multicellular host organisms.

  7. Wolbachia in guilds of Anastrepha fruit flies (Tephritidae) and parasitoid wasps (Braconidae)

    PubMed Central

    Mascarenhas, Rodrigo O; Prezotto, Leandro F; Perondini, André Luiz P; Marino, Celso Luiz; Selivon, Denise

    2016-01-01

    Abstract The endosymbiont Wolbachia is efficiently transmitted from females to their progenies, but horizontal transmission between different taxa is also known to occur. Aiming to determine if horizontal transmission might have occurred between Anastrepha fruit flies and associated braconid wasps, infection by Wolbachia was screened by amplification of a fragment of the wsp gene. Eight species of the genus Anastrepha were analyzed, from which six species of associated parasitoid wasps were recovered. The endosymbiont was found in seven Anastrepha species and in five species of braconids. The WSP Typing methodology detected eight wsp alleles belonging to Wolbachia supergroup A. Three were already known and five were new ones, among which four were found to be putative recombinant haplotypes. Two samples of Anastrepha obliqua and one sample of Doryctobracon brasiliensis showed multiple infection. Single infection by Wolbachia was found in the majority of samples. The distribution of Wolbachia harboring distinct alleles differed significantly between fruit flies and wasps. However, in nine samples of fruit flies and associated wasps, Wolbachia harbored the same wsp allele. These congruences suggest that horizontal transfer of Wolbachia might have occurred in the communities of fruit flies and their braconid parasitoids. PMID:27648768

  8. Unravelling the Wolbachia evolutionary role: the reprogramming of the host genomic imprinting.

    PubMed

    Negri, Ilaria; Franchini, Antonella; Gonella, Elena; Daffonchio, Daniele; Mazzoglio, Peter John; Mandrioli, Mauro; Alma, Alberto

    2009-07-07

    Environmental factors can induce significant epigenetic changes that may also be inherited by future generations. The maternally inherited symbiont of arthropods Wolbachia pipientis is an excellent candidate as an 'environmental' factor promoting trans-generational epigenetic changes: by establishing intimate relationships with germ-line cells, epigenetic effects of Wolbachia symbiosis would be manifested as a 'maternal effect', in which infection of the mother modulates the offspring phenotype. In the leafhopper Zyginidia pullula, Wolbachia feminizes genetic males, leaving them as intersexes. With the exception of male chitinous structures that are present in the last abdominal segment, feminized males display phenotypic features that are typical of females. These include ovaries that range from a typical histological architecture to an altered structure. Methylation-sensitive random amplification of polymorphic DNA profiles show that they possess a female genomic imprint. On the other hand, some rare feminized males bear testes instead of ovaries. These specimens possess a Wolbachia density approximately four orders of magnitude lower than feminized males with ovaries and maintain a male genome-methylation pattern. Our results indicate that Wolbachia infection disrupts male imprinting, which dramatically influences the expression of genes involved in sex differentiation and development, and the alteration occurs only if Wolbachia exceeds a density threshold. Thus, a new Wolbachia's role as an environmental evolutionary force, inducing epigenetic trans-generational changes, should now be considered.

  9. Wolbachia Influences the Production of Octopamine and Affects Drosophila Male Aggression

    PubMed Central

    Rohrscheib, Chelsie E.; Bondy, Elizabeth; Josh, Peter; Riegler, Markus; Eyles, Darryl; van Swinderen, Bruno; Weible, Michael W.

    2015-01-01

    Wolbachia bacteria are endosymbionts that infect approximately 40% of all insect species and are best known for their ability to manipulate host reproductive systems. Though the effect Wolbachia infection has on somatic tissues is less well understood, when present in cells of the adult Drosophila melanogaster brain, Wolbachia exerts an influence over behaviors related to olfaction. Here, we show that a strain of Wolbachia influences male aggression in flies, which is critically important in mate competition. A specific strain of Wolbachia was observed to reduce the initiation of aggressive encounters in Drosophila males compared to the behavior of their uninfected controls. To determine how Wolbachia was able to alter aggressive behavior, we investigated the role of octopamine, a neurotransmitter known to influence male aggressive behavior in many insect species. Transcriptional analysis of the octopamine biosynthesis pathway revealed that two essential genes, the tyrosine decarboxylase and tyramine β-hydroxylase genes, were significantly downregulated in Wolbachia-infected flies. Quantitative chemical analysis also showed that total octopamine levels were significantly reduced in the adult heads. PMID:25934616

  10. Wolbachia infection and the expression of cytoplasmic incompatibility in sandflies (Diptera: Psychodidae) from Egypt.

    PubMed

    Kassem, H A; Hassan, A N; Abdel-Hamid, I; Osman, G; El Khalab, E M; Madkour, M A

    2003-09-01

    A PCR-based method was used to screen four laboratory colonies of sandflies for Wolbachia infection. The colonies - one of Phlebotomus langeroni, one of P. bergeroti and two of P. papatasi - were all derived from sandflies collected in Egypt. Only one of the colonies, derived from P. papatasi collected in Sinai, was found infected. The sequence of the PCR product for this colony was identical to that previously reported for the Wolbachia in P. papatasi from Israel. The induction with tetracycline of cytoplasmic incompatibility (CI) in flies from the P. papatasi (Sinai) colony was then investigated, through reciprocal crosses between treated and untreated P. papatasi siblings. Partial CI expression was attained in the crosses involving antibiotic-treated (i.e. uninfected) females, whether the males used were infected with Wolbachia or had also been cleared of Wolbachia by antibiotic treatment. Most (75%) of the eggs oviposited by uninfected females that had been crossed with infected males, and most (58%) of those laid by uninfected females that had been crossed with uninfected males, failed to hatch. These results provide the first published evidence showing that Wolbachia infection in sandflies is advantageous to the insects. The failure to detect Wolbachia in one of the colonies derived from Egyptian P. papatasi or in the colonies derived from Egyptian P. bergeroti and P. langeroni may indicate that the inter- and intra-specific spread of Wolbachia is discontinuous, even within one country.

  11. Heat Sensitivity of wMel Wolbachia during Aedes aegypti Development.

    PubMed

    Ulrich, Jill N; Beier, John C; Devine, Gregor J; Hugo, Leon E

    2016-07-01

    The wMel strain of Wolbachia bacteria is known to prevent dengue and Zika virus transmission in the mosquito vector Aedes aegypti. Accordingly, the release of wMel-infected A. aegypti in endemic regions has been recommended by the World Health Organization as a potential strategy for controlling dengue and Zika outbreaks. However, the utility of this approach could be limited if high temperatures in the aquatic habitats where A. aegypti develop are detrimental to Wolbachia. We exposed wMel-infected A. aegypti eggs and larvae to fluctuating daily temperatures of 30-40°C for three, five, or seven days during their development. We found that Wolbachia levels in females emerging from heat treatments were significantly lower than in the controls that had developed at 20-30°C. Notably, seven days of high temperatures starting at the egg stage reduced Wolbachia levels in emerging females to less than 0.1% of the wMel control levels. However, after adult females returned to 20-30°C for 4-7 days, they experienced differing degrees of Wolbachia recovery. Our findings suggest that the spread of Wolbachia in wild A. aegypti populations and any consequent protection from dengue and Zika viruses might be limited in ecosystems that experience periods of extreme heat, but Wolbachia levels recover partially after temperatures return to normal.

  12. Molecular characterization of Wolbachia infection in bed bugs (Cimex lectularius) collected from several localities in France.

    PubMed

    Akhoundi, Mohammad; Cannet, Arnaud; Loubatier, Céline; Berenger, Jean-Michel; Izri, Arezki; Marty, Pierre; Delaunay, Pascal

    2016-01-01

    Wolbachia symbionts are maternally inherited intracellular bacteria that have been detected in numerous insects including bed bugs. The objective of this study, the first epidemiological study in Europe, was to screen Wolbachia infection among Cimex lectularius collected in the field, using PCR targeting the surface protein gene (wsp), and to compare obtained Wolbachia strains with those reported from laboratory colonies of C. lectularius as well as other Wolbachia groups. For this purpose, 284 bed bug specimens were caught and studied from eight different regions of France including the suburbs of Paris, Bouches-du-Rhône, Lot-et-Garonne, and five localities in Alpes-Maritimes. Among the samples, 166 were adults and the remaining 118 were considered nymphs. In all, 47 out of 118 nymphs (40%) and 61 out of 166 adults (37%) were found positive on wsp screening. Among the positive cases, 10 samples were selected randomly for sequencing. The sequences had 100% homology with wsp sequences belonging to the F-supergroup strains of Wolbachia. Therefore, we confirm the similarity of Wolbachia strains detected in this epidemiological study to Wolbachia spp. reported from laboratory colonies of C. lectularius.

  13. Wolbachia Influences the Maternal Transmission of the gypsy Endogenous Retrovirus in Drosophila melanogaster

    PubMed Central

    Touret, Franck; Guiguen, François

    2014-01-01

    ABSTRACT The endosymbiotic bacteria of the genus Wolbachia are present in most insects and are maternally transmitted through the germline. Moreover, these intracellular bacteria exert antiviral activity against insect RNA viruses, as in Drosophila melanogaster, which could explain the prevalence of Wolbachia bacteria in natural populations. Wolbachia is maternally transmitted in D. melanogaster through a mechanism that involves distribution at the posterior pole of mature oocytes and then incorporation into the pole cells of the embryos. In parallel, maternal transmission of several endogenous retroviruses is well documented in D. melanogaster. Notably, gypsy retrovirus is expressed in permissive follicle cells and transferred to the oocyte and then to the offspring by integrating into their genomes. Here, we show that the presence of Wolbachia wMel reduces the rate of gypsy insertion into the ovo gene. However, the presence of Wolbachia does not modify the expression levels of gypsy RNA and envelope glycoprotein from either permissive or restrictive ovaries. Moreover, Wolbachia affects the pattern of distribution of the retroviral particles and the gypsy envelope protein in permissive follicle cells. Altogether, our results enlarge the knowledge of the antiviral activity of Wolbachia to include reducing the maternal transmission of endogenous retroviruses in D. melanogaster. PMID:25182324

  14. Supergroup C Wolbachia, mutualist symbionts of filarial nematodes, have a distinct genome structure

    PubMed Central

    Comandatore, Francesco; Cordaux, Richard; Bandi, Claudio; Blaxter, Mark; Darby, Alistair; Makepeace, Benjamin L.; Montagna, Matteo; Sassera, Davide

    2015-01-01

    Wolbachia pipientis is possibly the most widespread endosymbiont of arthropods and nematodes. While all Wolbachia strains have historically been defined as a single species, 16 monophyletic clusters of diversity (called supergroups) have been described. Different supergroups have distinct host ranges and symbiotic relationships, ranging from mutualism to reproductive manipulation. In filarial nematodes, which include parasites responsible for major diseases of humans (such as Onchocerca volvulus, agent of river blindness) and companion animals (Dirofilaria immitis, the dog heartworm), Wolbachia has an obligate mutualist role and is the target of new treatment regimens. Here, we compare the genomes of eight Wolbachia strains, spanning the diversity of the major supergroups (A–F), analysing synteny, transposable element content, GC skew and gene loss or gain. We detected genomic features that differ between Wolbachia supergroups, most notably in the C and D clades from filarial nematodes. In particular, strains from supergroup C (symbionts of O. volvulus and D. immitis) present a pattern of GC skew, conserved synteny and lack of transposable elements, unique in the Wolbachia genus. These features could be the consequence of a distinct symbiotic relationship between C Wolbachia strains and their hosts, highlighting underappreciated differences between the mutualistic supergroups found within filarial nematodes. PMID:26631376

  15. Evidence for a new feminizing Wolbachia strain in the isopod Armadillidium vulgare: evolutionary implications.

    PubMed

    Cordaux, R; Michel-Salzat, A; Frelon-Raimond, M; Rigaud, T; Bouchon, D

    2004-07-01

    Wolbachia are intracellular maternally inherited alpha-Proteobacteria infecting a wide range of arthropods. In the common pill bug Armadillidium vulgare, the known Wolbachia strain is responsible for feminization of genetic males. We have investigated Wolbachia diversity in 20 populations of A. vulgare from west and east Europe, north Africa and north America. A new Wolbachia strain (wVulM) was identified through the variability of the wsp gene, distantly related to that previously known (wVulC) in this host species. No individual with multiple infections was detected. Inoculation experiments indicated that the new wVulM bacterial strain also induces feminization in A. vulgare. However, the wVulC strain showed a higher transmission rate than the wVulM strain and was the most geographically widespread Wolbachia in A. vulgare populations. Mitochondrial 16SrDNA gene sequencing was conducted in Wolbachia-infected individuals, revealing the occurrence of four host lineages. The comparison of bacterial strains and their respective host mitochondrial phylogenies failed to show concordance, indicating horizontal transmission of the Wolbachia strains within populations of A. vulgare.

  16. Heat Sensitivity of wMel Wolbachia during Aedes aegypti Development

    PubMed Central

    Beier, John C.; Devine, Gregor J.; Hugo, Leon E.

    2016-01-01

    The wMel strain of Wolbachia bacteria is known to prevent dengue and Zika virus transmission in the mosquito vector Aedes aegypti. Accordingly, the release of wMel-infected A. aegypti in endemic regions has been recommended by the World Health Organization as a potential strategy for controlling dengue and Zika outbreaks. However, the utility of this approach could be limited if high temperatures in the aquatic habitats where A. aegypti develop are detrimental to Wolbachia. We exposed wMel-infected A. aegypti eggs and larvae to fluctuating daily temperatures of 30–40°C for three, five, or seven days during their development. We found that Wolbachia levels in females emerging from heat treatments were significantly lower than in the controls that had developed at 20–30°C. Notably, seven days of high temperatures starting at the egg stage reduced Wolbachia levels in emerging females to less than 0.1% of the wMel control levels. However, after adult females returned to 20–30°C for 4–7 days, they experienced differing degrees of Wolbachia recovery. Our findings suggest that the spread of Wolbachia in wild A. aegypti populations and any consequent protection from dengue and Zika viruses might be limited in ecosystems that experience periods of extreme heat, but Wolbachia levels recover partially after temperatures return to normal. PMID:27459519

  17. Comparative analysis of wolbachia genomes reveals streamlining and divergence of minimalist two-component systems.

    PubMed

    Christensen, Steen; Serbus, Laura Renee

    2015-03-24

    Two-component regulatory systems are commonly used by bacteria to coordinate intracellular responses with environmental cues. These systems are composed of functional protein pairs consisting of a sensor histidine kinase and cognate response regulator. In contrast to the well-studied Caulobacter crescentus system, which carries dozens of these pairs, the streamlined bacterial endosymbiont Wolbachia pipientis encodes only two pairs: CckA/CtrA and PleC/PleD. Here, we used bioinformatic tools to compare characterized two-component system relays from C. crescentus, the related Anaplasmataceae species Anaplasma phagocytophilum and Ehrlichia chaffeensis, and 12 sequenced Wolbachia strains. We found the core protein pairs and a subset of interacting partners to be highly conserved within Wolbachia and these other Anaplasmataceae. Genes involved in two-component signaling were positioned differently within the various Wolbachia genomes, whereas the local context of each gene was conserved. Unlike Anaplasma and Ehrlichia, Wolbachia two-component genes were more consistently found clustered with metabolic genes. The domain architecture and key functional residues standard for two-component system proteins were well-conserved in Wolbachia, although residues that specify cognate pairing diverged substantially from other Anaplasmataceae. These findings indicate that Wolbachia two-component signaling pairs share considerable functional overlap with other α-proteobacterial systems, whereas their divergence suggests the potential for regulatory differences and cross-talk.

  18. Supergroup C Wolbachia, mutualist symbionts of filarial nematodes, have a distinct genome structure.

    PubMed

    Comandatore, Francesco; Cordaux, Richard; Bandi, Claudio; Blaxter, Mark; Darby, Alistair; Makepeace, Benjamin L; Montagna, Matteo; Sassera, Davide

    2015-12-01

    Wolbachia pipientis is possibly the most widespread endosymbiont of arthropods and nematodes. While all Wolbachia strains have historically been defined as a single species, 16 monophyletic clusters of diversity (called supergroups) have been described. Different supergroups have distinct host ranges and symbiotic relationships, ranging from mutualism to reproductive manipulation. In filarial nematodes, which include parasites responsible for major diseases of humans (such as Onchocerca volvulus, agent of river blindness) and companion animals (Dirofilaria immitis, the dog heartworm), Wolbachia has an obligate mutualist role and is the target of new treatment regimens. Here, we compare the genomes of eight Wolbachia strains, spanning the diversity of the major supergroups (A-F), analysing synteny, transposable element content, GC skew and gene loss or gain. We detected genomic features that differ between Wolbachia supergroups, most notably in the C and D clades from filarial nematodes. In particular, strains from supergroup C (symbionts of O. volvulus and D. immitis) present a pattern of GC skew, conserved synteny and lack of transposable elements, unique in the Wolbachia genus. These features could be the consequence of a distinct symbiotic relationship between C Wolbachia strains and their hosts, highlighting underappreciated differences between the mutualistic supergroups found within filarial nematodes.

  19. The impacts of Wolbachia and the microbiome on mate choice in Drosophila melanogaster.

    PubMed

    Arbuthnott, D; Levin, T C; Promislow, D E L

    2016-02-01

    Symbionts and parasites can manipulate their hosts' reproduction to their own benefit, profoundly influencing patterns of mate choice and evolution of the host population. Wolbachia is one of the most widespread symbionts among arthropods, and one that alters its hosts' reproduction in diverse and dramatic ways. While we are beginning to appreciate how Wolbachia's extreme manipulations of host reproduction can influence species diversification and reproductive isolation, we understand little about how symbionts and Wolbachia, in particular, may affect intrapopulation processes of mate choice. We hypothesized that the maternally transmitted Wolbachia would increase the attractiveness of its female hosts to further its own spread. We therefore tested the effects of Wolbachia removal and microbiome disruption on female attractiveness and male mate choice among ten isofemale lines of Drosophila melanogaster. We found variable effects of general microbiome disruption on female attractiveness, with indications that bacteria interact with hosts in a line-specific manner to affect female attractiveness. However, we found no evidence that Wolbachia influence female attractiveness or male mate choice among these lines. Although the endosymbiont Wolbachia can greatly alter the reproduction of their hosts in many species, there is no indication that they alter mate choice behaviours in D. melanogaster.

  20. Wolbachia Influences the Production of Octopamine and Affects Drosophila Male Aggression.

    PubMed

    Rohrscheib, Chelsie E; Bondy, Elizabeth; Josh, Peter; Riegler, Markus; Eyles, Darryl; van Swinderen, Bruno; Weible, Michael W; Brownlie, Jeremy C

    2015-07-01

    Wolbachia bacteria are endosymbionts that infect approximately 40% of all insect species and are best known for their ability to manipulate host reproductive systems. Though the effect Wolbachia infection has on somatic tissues is less well understood, when present in cells of the adult Drosophila melanogaster brain, Wolbachia exerts an influence over behaviors related to olfaction. Here, we show that a strain of Wolbachia influences male aggression in flies, which is critically important in mate competition. A specific strain of Wolbachia was observed to reduce the initiation of aggressive encounters in Drosophila males compared to the behavior of their uninfected controls. To determine how Wolbachia was able to alter aggressive behavior, we investigated the role of octopamine, a neurotransmitter known to influence male aggressive behavior in many insect species. Transcriptional analysis of the octopamine biosynthesis pathway revealed that two essential genes, the tyrosine decarboxylase and tyramine β-hydroxylase genes, were significantly downregulated in Wolbachia-infected flies. Quantitative chemical analysis also showed that total octopamine levels were significantly reduced in the adult heads.

  1. Discovery of putative small non-coding RNAs from the obligate intracellular bacterium Wolbachia pipientis.

    PubMed

    Woolfit, Megan; Algama, Manjula; Keith, Jonathan M; McGraw, Elizabeth A; Popovici, Jean

    2015-01-01

    Wolbachia pipientis is an endosymbiotic bacterium that induces a wide range of effects in its insect hosts, including manipulation of reproduction and protection against pathogens. Little is known of the molecular mechanisms underlying the insect-Wolbachia interaction, though it is likely to be mediated via the secretion of proteins or other factors. There is an increasing amount of evidence that bacteria regulate many cellular processes, including secretion of virulence factors, using small non-coding RNAs (sRNAs), but sRNAs have not previously been described from Wolbachia. We have used two independent approaches, one based on comparative genomics and the other using RNA-Seq data generated for gene expression studies, to identify candidate sRNAs in Wolbachia. We experimentally characterized the expression of one of these candidates in four Wolbachia strains, and showed that it is differentially regulated in different host tissues and sexes. Given the roles played by sRNAs in other host-associated bacteria, the conservation of the candidate sRNAs between different Wolbachia strains, and the sex- and tissue-specific differential regulation we have identified, we hypothesise that sRNAs may play a significant role in the biology of Wolbachia, and in particular in its interactions with its host.

  2. Wolbachia infections in world populations of bean beetles (Coleoptera: Chrysomelidae: Bruchinae) infesting cultivated and wild legumes.

    PubMed

    Kondo, Natsuko I; Tuda, Midori; Toquenaga, Yukihiko; Lan, Yen-Chiu; Buranapanichpan, Sawai; Horng, Shwu-Bin; Shimada, Masakazu; Fukatsu, Takema

    2011-07-01

    Wolbachia endosymbionts are widespread among insects and other arthropods, often causing cytoplasmic incompatibility and other reproductive phenotypes in their hosts. Recently, possibilities of Wolbachia-mediated pest control and management have been proposed, and the bean beetles of the subfamily Bruchinae are known as serious pests of harvested and stored beans worldwide. Here we investigated Wolbachia infections in bean beetles from the world, representing seven genera, 20 species and 87 populations. Of 20 species examined, Wolbachia infections were detected in four species, Megabruchidius sophorae, Callosobruchus analis, C. latealbus and C. chinensis. Infection frequencies were partial in M. sophorae but perfect in the other species. In addition to C. chinensis described in the previous studies, C. latealbus was infected with two distinct Wolbachia strains. These Wolbachia strains from the bean beetles were phylogenetically not closely related to each other. Among world populations of C. chinensis, some Taiwanese populations on a wild leguminous plant, Rhynchosia minima, exhibited a peculiar Wolbachia infection pattern, suggesting the possibility that these populations comprise a distinct host race or a cryptic species.

  3. Presence of Extensive Wolbachia Symbiont Insertions Discovered in the Genome of Its Host Glossina morsitans morsitans

    PubMed Central

    Falchetto, Marco; Gomulski, Ludvik M.; Telleria, Erich; Alam, Uzma; Doudoumis, Vangelis; Scolari, Francesca; Benoit, Joshua B.; Swain, Martin; Takac, Peter; Malacrida, Anna R.; Bourtzis, Kostas; Aksoy, Serap

    2014-01-01

    Tsetse flies (Glossina spp.) are the cyclical vectors of Trypanosoma spp., which are unicellular parasites responsible for multiple diseases, including nagana in livestock and sleeping sickness in humans in Africa. Glossina species, including Glossina morsitans morsitans (Gmm), for which the Whole Genome Sequence (WGS) is now available, have established symbiotic associations with three endosymbionts: Wigglesworthia glossinidia, Sodalis glossinidius and Wolbachia pipientis (Wolbachia). The presence of Wolbachia in both natural and laboratory populations of Glossina species, including the presence of horizontal gene transfer (HGT) events in a laboratory colony of Gmm, has already been shown. We herein report on the draft genome sequence of the cytoplasmic Wolbachia endosymbiont (cytWol) associated with Gmm. By in silico and molecular and cytogenetic analysis, we discovered and validated the presence of multiple insertions of Wolbachia (chrWol) in the host Gmm genome. We identified at least two large insertions of chrWol, 527,507 and 484,123 bp in size, from Gmm WGS data. Southern hybridizations confirmed the presence of Wolbachia insertions in Gmm genome, and FISH revealed multiple insertions located on the two sex chromosomes (X and Y), as well as on the supernumerary B-chromosomes. We compare the chrWol insertions to the cytWol draft genome in an attempt to clarify the evolutionary history of the HGT events. We discuss our findings in light of the evolution of Wolbachia infections in the tsetse fly and their potential impacts on the control of tsetse populations and trypanosomiasis. PMID:24763283

  4. Fitness advantage and cytoplasmic incompatibility in Wolbachia single- and superinfected Aedes albopictus.

    PubMed

    Dobson, S L; Rattanadechakul, W; Marsland, E J

    2004-08-01

    Wolbachia are obligate, maternally inherited, intracellular bacteria that infect numerous insects and other invertebrates. Wolbachia infections have evolved multiple mechanisms to manipulate host reproduction and facilitate invasion of naive host populations. One such mechanism is cytoplasmic incompatibility (CI) that occurs in many insect species, including Aedes albopictus (Asian tiger mosquito). The multiple Wolbachia infections that occur naturally in A. albopictus make this mosquito a useful system in which to study CI. Here, experiments employ mosquito strains that have been introgressed to provide genetically similar strains that harbor differing Wolbachia infection types. Cytoplasmic incompatibility levels, host longevity, egg hatch rates, and fecundity are examined. Crossing results demonstrate a pattern of additive unidirectional cytoplasmic incompatibility. Furthermore, relative to uninfected females, infected females are at a reproductive advantage due to both cytoplasmic incompatibility and a fitness increase associated with Wolbachia infection. In contrast, no fitness difference was observed in comparisons of single- and superinfected females. We discuss the observed results in regard to the evolution of the Wolbachia/A. albopictus symbiosis and the observed pattern of Wolbachia infection in natural populations.

  5. Wolbachia association with the tsetse fly, Glossina fuscipes fuscipes, reveals high levels of genetic diversity and complex evolutionary dynamics

    PubMed Central

    2013-01-01

    Background Wolbachia pipientis, a diverse group of α-proteobacteria, can alter arthropod host reproduction and confer a reproductive advantage to Wolbachia-infected females (cytoplasmic incompatibility (CI)). This advantage can alter host population genetics because Wolbachia-infected females produce more offspring with their own mitochondrial DNA (mtDNA) haplotypes than uninfected females. Thus, these host haplotypes become common or fixed (selective sweep). Although simulations suggest that for a CI-mediated sweep to occur, there must be a transient phase with repeated initial infections of multiple individual hosts by different Wolbachia strains, this has not been observed empirically. Wolbachia has been found in the tsetse fly, Glossina fuscipes fuscipes, but it is not limited to a single host haplotype, suggesting that CI did not impact its population structure. However, host population genetic differentiation could have been generated if multiple Wolbachia strains interacted in some populations. Here, we investigated Wolbachia genetic variation in G. f. fuscipes populations of known host genetic composition in Uganda. We tested for the presence of multiple Wolbachia strains using Multi-Locus Sequence Typing (MLST) and for an association between geographic region and host mtDNA haplotype using Wolbachia DNA sequence from a variable locus, groEL (heat shock protein 60). Results MLST demonstrated that some G. f. fuscipes carry Wolbachia strains from two lineages. GroEL revealed high levels of sequence diversity within and between individuals (Haplotype diversity = 0.945). We found Wolbachia associated with 26 host mtDNA haplotypes, an unprecedented result. We observed a geographical association of one Wolbachia lineage with southern host mtDNA haplotypes, but it was non-significant (p = 0.16). Though most Wolbachia-infected host haplotypes were those found in the contact region between host mtDNA groups, this association was non-significant (p = 0

  6. Localization of a filarial phosphate permease that is up-regulated in response to depletion of essential Wolbachia endobacteria.

    PubMed

    Arumugam, Sridhar; Hoerauf, Achim; Pfarr, Kenneth M

    2014-03-01

    Wolbachia of filarial nematodes are essential, obligate endobacteria. When depleted by doxycycline worm embryogenesis, larval development and worm survival are inhibited. The molecular basis governing the endosymbiosis between Wolbachia and their filarial host is still being deciphered. In rodent filarial nematode Litomosoides sigmodontis, a nematode encoded phosphate permease gene (Ls-ppe-1) was up-regulated at the mRNA level in response to Wolbachia depletion and this gene promises to have an important role in Wolbachia-nematode endosymbiosis. To further characterize this gene, the regulation of phosphate permease during Wolbachia depletion was studied at the protein level in L. sigmodontis and in the human filaria Onchocerca volvulus. And the localization of phosphate permease (PPE) and Wolbachia in L. sigmodontis and O. volvulus was investigated in untreated and antibiotic treated worms. Depletion of Wolbachia by tetracycline (Tet) resulted in up-regulation of Ls-ppe-1 in L. sigmodontis. On day 36 of Tet treatment, compared to controls (Con), >98% of Wolbachia were depleted with a 3-fold increase in mRNA levels of Ls-ppe-1. Anti-Ls-PPE serum used in Western blots showed up-regulation of Ls-PPE at the protein level in Tet worms on day 15 and 36 of treatment. Immunohistology revealed the localization of Wolbachia and Ls-PPE in the embryos, microfilariae and hypodermis of L. sigmodontis female worms and up-regulation of Ls-PPE in response to Wolbachia depletion. Expression of O. volvulus phosphate permease (Ov-PPE) studied using anti-Ov-PPE serum, showed up-regulation of Ov-PPE at the protein level in doxycycline treated Wolbachia depleted O. volvulus worms and immunohistology revealed localization of Ov-PPE and Wolbachia and up-regulation of Ov-PPE in the hypodermis and embryos of doxycycline treated worms. Ls-PPE and Ov-PPE are upregulated upon Wolbachia depletion in same tissues and regions where Wolbachia are located in untreated worms, reinforcing a link

  7. 46 CFR 122.515 - Passenger safety bill.

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    ... 46 Shipping 4 2014-10-01 2014-10-01 false Passenger safety bill. 122.515 Section 122.515 Shipping... Emergencies § 122.515 Passenger safety bill. (a) A passenger safety bill must be posted by the master in each... accommodations for more than 49 passengers. (b) Each passenger safety bill required by this section must list:...

  8. 46 CFR 122.515 - Passenger safety bill.

    Code of Federal Regulations, 2012 CFR

    2012-10-01

    ... 46 Shipping 4 2012-10-01 2012-10-01 false Passenger safety bill. 122.515 Section 122.515 Shipping... Emergencies § 122.515 Passenger safety bill. (a) A passenger safety bill must be posted by the master in each... accommodations for more than 49 passengers. (b) Each passenger safety bill required by this section must list:...

  9. Variations in the sensitivity of different primers for detecting Wolbachia in Anastrepha (diptera: tephritidae)

    PubMed Central

    Marcon, Helena Sanches; Coscrato, Virgínia Elias; Selivon, Denise; Perondini, André Luiz Paranhos; Marino, Celso Luis

    2011-01-01

    Wolbachia are endosymbiont bacteria of the family Rickettsiacea that are widespread in invertebrates and occur between 20% and 60% of Neotropical insects. These bacteria are responsible for reproductive phenomena such as cytoplasmic incompatibility, male killing, feminization and parthenogenesis. Supergroups A and B of Wolbachia are common in insects and can be identified using primers for 16S rDNA, ftsZ and wsp; these primers vary in their ability to detect Wolbachia. The ftsZ primer was the first primer used to detect Wolbachia in Anastrepha fruit flies. The primers for 16S rDNA, ftsZ and wsp and the corresponding PCR conditions have been optimized to study the distribution of Wolbachia and their effect on the biology of Anastrepha in Brazil. In this work, we examined the ability of these primers to detect Wolbachia in Anastrepha populations from three regions in the State of São Paulo, southeastern Brazil. All of the samples were positive for Wolbachia supergroup A when screened with primers for 16S A rDNA and wsp A; the wsp B primer also gave a positive result, indicating cross-reactivity. The ftsZ primer showed a poor ability to detect Wolbachia in Anastrepha and generated false negatives in 44.9% of the samples. These findings indicate that reliable PCR detection of Wolbachia requires the use of primers for 16S rDNA and wsp to avoid cross-reactions and false negatives, and that the ftsZ primer needs to be redesigned to improve its selectivity. PMID:24031693

  10. Mechanisms of Horizontal Cell-to-Cell Transfer of Wolbachia spp. in Drosophila melanogaster.

    PubMed

    White, Pamela M; Pietri, Jose E; Debec, Alain; Russell, Shelbi; Patel, Bhavin; Sullivan, William

    2017-04-01

    Wolbachia is an intracellular endosymbiont present in most arthropod and filarial nematode species. Transmission between hosts is primarily vertical, taking place exclusively through the female germ line, although horizontal transmission has also been documented. The results of several studies indicate that Wolbachia spp. can undergo transfer between somatic and germ line cells during nematode development and in adult flies. However, the mechanisms underlying horizontal cell-to-cell transfer remain largely unexplored. Here, we establish a tractable system for probing horizontal transfer of Wolbachia cells between Drosophila melanogaster cells in culture using fluorescence in situ hybridization (FISH). First, we show that horizontal transfer is independent of cell-to-cell contact and can efficiently take place through the culture medium within hours. Further, we demonstrate that efficient transfer utilizes host cell phagocytic and clathrin/dynamin-dependent endocytic machinery. Lastly, we provide evidence that this process is conserved between species, showing that horizontal transfer from mosquito to Drosophila cells takes place in a similar fashion. Altogether, our results indicate that Wolbachia utilizes host internalization machinery during infection, and this mechanism is conserved across insect species.IMPORTANCE Our work has broad implications for the control and treatment of tropical diseases. Wolbachia can confer resistance against a variety of human pathogens in mosquito vectors. Elucidating the mechanisms of horizontal transfer will be useful for efforts to more efficiently infect nonnatural insect hosts with Wolbachia as a biological control agent. Further, as Wolbachia is essential for the survival of filarial nematodes, understanding horizontal transfer might provide new approaches to treating human infections by targeting Wolbachia Finally, this work provides a key first step toward the genetic manipulation of Wolbachia.

  11. Wolbachia-Based Dengue Virus Inhibition Is Not Tissue-Specific in Aedes aegypti

    PubMed Central

    Amuzu, Hilaria E.; McGraw, Elizabeth A.

    2016-01-01

    Background Dengue fever, caused by the dengue virus (DENV), is now the most common arbovirus transmitted disease globally. One novel approach to control DENV is to use the endosymbiotic bacterium, Wolbachia pipientis, to limit DENV replication inside the primary mosquito vector, Aedes aegypti. Wolbachia that is naturally present in a range of insects reduces the capacity for viruses, bacteria, parasites and fungi to replicate inside insects. Wolbachia’s mode of action is not well understood but may involve components of immune activation or competition with pathogens for limited host resources. The strength of Wolbachia-based anti DENV effects appear to correlate with bacterial density in the whole insect and in cell culture. Here we aimed to determine whether particular tissues, especially those with high Wolbachia densities or immune activity, play a greater role in mediating the anti DENV effect. Methodology/findings Ae. aegypti mosquito lines with and without Wolbachia (Wildtype) were orally fed DENV 3 and their viral loads subsequently measured over two time points post infection in the midgut, head, salivary glands, Malpighian tubules, fat body and carcass. We did not find correlations between Wolbachia densities and DENV loads in any tissue, nor with DENV loads in salivary glands, the endpoint of infection. This is in contrast with strong positive correlations between DENV loads in a range of tissues and salivary gland loads for Wildtype mosquitoes. Lastly, there was no evidence of a heightened role for tissues with known immune function including the fat body and the Malpighian tubules in Wolbachia’s limitation of DENV. Conclusion/significance We conclude that the efficacy of DENV blocking in Wolbachia infected mosquitoes is not reliant on any particular tissue. This work therefore suggests that the mechanism of Wolbachia-based antiviral effects is either systemic or acts locally via processes that are fundamental to diverse cell types. We further

  12. Wolbachia infection status and genetic structure in natural populations of Polytremis nascens (Lepidoptera: Hesperiidae).

    PubMed

    Jiang, Weibin; Zhu, Jianqing; Chen, Minghan; Yang, Qichang; Du, Xuan; Chen, Shiyan; Zhang, Lina; Yu, Yiming; Yu, Weidong

    2014-10-01

    The maternally inherited obligate bacteria Wolbachia is known for infecting the reproductive tissues of a wide range of arthropods. In this study, we surveyed Wolbachia infections in Polytremis nascens (Lepidoptera: Hesperiidae) from 14 locations in China by amplifying the 16S rRNA gene with a nested PCR method and revealed the effect of Wolbachia on host mitochondrial DNA. The results show that 31% (21/67) are Wolbachia positive among all specimens and mainly prevails in southern populations in China. No significant difference in the prevalence is found between the sexes. Notably, the nucleotide diversity of Wolbachia infected butterflies is smaller compared to that of uninfected butterflies. The mitochondrial DNA of infected group appear to be not evolving neutrally (Tajima's D value=-2.3303 and Fu's F values=-3.7068). The analysis of molecular variance shows significant differentiation of mitochondrial haplotypes between infected and uninfected specimens (FST=0.6064). The mismatch analysis speculated the different expansion pattern in Wolbachia infected specimens and all P. nascens specimens. These results suggest that the populations of P. nascens may have recently been subjected to a Wolbachia-induced sweep. Additionally, phylogenetic analysis differentiated the mitochondrial haplotypes of P. nascens into three major clades. The clades are in perfect agreement with the pattern of Wolbachia infection. One of the clades grouped with the butterflies infected with Wolbachia. The remaining two clades grouped with uninfected butterflies from the central-west of China populations and Eastern and Southern China populations respectively, which are isolated mainly by the Yangtze River. The analysis of haplotype networks, geographic distribution and population size change shows that Haplotype 1 in central-west of China is the ancestral haplotype and the populations of P. nascens are expanded.

  13. Wolbachia infections that reduce immature insect survival: Predicted impacts on population replacement

    PubMed Central

    2011-01-01

    Background The evolutionary success of Wolbachia bacteria, infections of which are widespread in invertebrates, is largely attributed to an ability to manipulate host reproduction without imposing substantial fitness costs. Here, we describe a stage-structured model with deterministic immature lifestages and a stochastic adult female lifestage. Simulations were conducted to better understand Wolbachia invasions into uninfected host populations. The model includes conventional Wolbachia parameters (the level of cytoplasmic incompatibility, maternal inheritance, the relative fecundity of infected females, and the initial Wolbachia infection frequency) and a new parameter termed relative larval viability (RLV), which is the survival of infected larvae relative to uninfected larvae. Results The results predict the RLV parameter to be the most important determinant for Wolbachia invasion and establishment. Specifically, the fitness of infected immature hosts must be close to equal to that of uninfected hosts before population replacement can occur. Furthermore, minute decreases in RLV inhibit the invasion of Wolbachia despite high levels of cytoplasmic incompatibility, maternal inheritance, and low adult fitness costs. Conclusions The model described here takes a novel approach to understanding the spread of Wolbachia through a population with explicit dynamics. By combining a stochastic female adult lifestage and deterministic immature/adult male lifestages, the model predicts that even those Wolbachia infections that cause minor decreases in immature survival are unlikely to invade and spread within the host population. The results are discussed in relation to recent theoretical and empirical studies of natural population replacement events and proposed applied research, which would use Wolbachia as a tool to manipulate insect populations. PMID:21975225

  14. Modelling the transmission dynamics of dengue in the presence of Wolbachia.

    PubMed

    Ndii, Meksianis Z; Hickson, R I; Allingham, David; Mercer, G N

    2015-04-01

    Use of the bacterium Wolbachia is an innovative new strategy designed to break the cycle of dengue transmission. There are two main mechanisms by which Wolbachia could achieve this: by reducing the level of dengue virus in the mosquito and/or by shortening the host mosquito's lifespan. However, although Wolbachia shortens the lifespan, it also gives a breeding advantage which results in complex population dynamics. This study focuses on the development of a mathematical model to quantify the effect on human dengue cases of introducing Wolbachia into the mosquito population. The model consists of a compartment-based system of first-order differential equations; seasonal forcing in the mosquito population is introduced through the adult mosquito death rate. The analysis focuses on a single dengue outbreak typical of a region with a strong seasonally-varying mosquito population. We found that a significant reduction in human dengue cases can be obtained provided that Wolbachia-carrying mosquitoes persist when competing with mosquitoes without Wolbachia. Furthermore, using the Wolbachia strain WMel reduces the mosquito lifespan by at most 10% and allows them to persist in competition with non-Wolbachia-carrying mosquitoes. Mosquitoes carrying the WMelPop strain, however, are not likely to persist as it reduces the mosquito lifespan by up to 50%. When all other effects of Wolbachia on the mosquito physiology are ignored, cytoplasmic incompatibility alone results in a reduction in the number of human dengue cases. A sensitivity analysis of the parameters in the model shows that the transmission probability, the biting rate and the average adult mosquito death rate are the most important parameters for the outcome of the cumulative proportion of human individuals infected with dengue.

  15. Costs of Three Wolbachia Infections on the Survival of Aedes aegypti Larvae under Starvation Conditions.

    PubMed

    Ross, Perran A; Endersby, Nancy M; Hoffmann, Ary A

    2016-01-01

    The mosquito Aedes aegypti, the principal vector of dengue virus, has recently been infected experimentally with Wolbachia: intracellular bacteria that possess potential as dengue biological control agents. Wolbachia depend on their hosts for nutrients they are unable to synthesize themselves. Consequently, competition between Wolbachia and their host for resources could reduce host fitness under the competitive conditions commonly experienced by larvae of Ae. aegypti in the field, hampering the invasion of Wolbachia into natural mosquito populations. We assess the survival and development of Ae. aegypti larvae under starvation conditions when infected with each of three experimentally-generated Wolbachia strains: wMel, wMelPop and wAlbB, and compare their fitness to wild-type uninfected larvae. We find that all three Wolbachia infections reduce the survival of larvae relative to those that are uninfected, and the severity of the effect is concordant with previously characterized fitness costs to other life stages. We also investigate the ability of larvae to recover from extended food deprivation and find no effect of Wolbachia on this trait. Aedes aegypti larvae of all infection types were able to resume their development after one month of no food, pupate rapidly, emerge at a large size, and exhibit complete cytoplasmic incompatibility and maternal transmission. A lowered ability of Wolbachia-infected larvae to survive under starvation conditions will increase the threshold infection frequency required for Wolbachia to establish in highly competitive natural Ae. aegypti populations and will also reduce the speed of invasion. This study also provides insights into survival strategies of larvae when developing in stressful environments.

  16. Extreme divergence of Wolbachia tropism for the stem-cell-niche in the Drosophila testis.

    PubMed

    Toomey, Michelle E; Frydman, Horacio M

    2014-12-01

    Microbial tropism, the infection of specific cells and tissues by a microorganism, is a fundamental aspect of host-microbe interactions. The intracellular bacteria Wolbachia have a peculiar tropism for the stem cell niches in the Drosophila ovary, the microenvironments that support the cells producing the eggs. The molecular underpinnings of Wolbachia stem cell niche tropism are unknown. We have previously shown that the patterns of tropism in the ovary show a high degree of conservation across the Wolbachia lineage, with closely related Wolbachia strains usually displaying the same pattern of stem cell niche tropism. It has also been shown that tropism to these structures in the ovary facilitates both vertical and horizontal transmission, providing a strong selective pressure towards evolutionary conservation of tropism. Here we show great disparity in the evolutionary conservation and underlying mechanisms of stem cell niche tropism between male and female gonads. In contrast to females, niche tropism in the male testis is not pervasive, present in only 45% of niches analyzed. The patterns of niche tropism in the testis are not evolutionarily maintained across the Wolbachia lineage, unlike what was shown in the females. Furthermore, hub tropism does not correlate with cytoplasmic incompatibility, a Wolbachia-driven phenotype imprinted during spermatogenesis. Towards identifying the molecular mechanism of hub tropism, we performed hybrid analyses of Wolbachia strains in non-native hosts. These results indicate that both Wolbachia and host derived factors play a role in the targeting of the stem cell niche in the testis. Surprisingly, even closely related Wolbachia strains in Drosophila melanogaster, derived from a single ancestor only 8,000 years ago, have significantly different tropisms to the hub, highlighting that stem cell niche tropism is rapidly diverging in males. These findings provide a powerful system to investigate the mechanisms and evolution of

  17. Wolbachia utilize host actin for efficient maternal transmission in Drosophila melanogaster.

    PubMed

    Newton, Irene L G; Savytskyy, Oleksandr; Sheehan, Kathy B

    2015-04-01

    Wolbachia pipientis is a ubiquitous, maternally transmitted bacterium that infects the germline of insect hosts. Estimates are that Wolbachia infect nearly 40% of insect species on the planet, making it the most prevalent infection on Earth. The bacterium, infamous for the reproductive phenotypes it induces in arthropod hosts, has risen to recent prominence due to its use in vector control. Wolbachia infection prevents the colonization of vectors by RNA viruses, including Drosophila C virus and important human pathogens such as Dengue and Chikungunya. Here we present data indicating that Wolbachia utilize the host actin cytoskeleton during oogenesis for persistence within and transmission between Drosophila melanogaster generations. We show that phenotypically wild type flies heterozygous for cytoskeletal mutations in Drosophila profilin (chic(221/+) and chic(1320/+)) or villin (qua(6-396/+)) either clear a Wolbachia infection, or result in significantly reduced infection levels. This reduction of Wolbachia is supported by PCR evidence, Western blot results and cytological examination. This phenotype is unlikely to be the result of maternal loading defects, defects in oocyte polarization, or germline stem cell proliferation, as the flies are phenotypically wild type in egg size, shape, and number. Importantly, however, heterozygous mutant flies exhibit decreased total G-actin in the ovary, compared to control flies and chic(221) heterozygous mutants exhibit decreased expression of profilin. Additionally, RNAi knockdown of profilin during development decreases Wolbachia titers. We analyze evidence in support of alternative theories to explain this Wolbachia phenotype and conclude that our results support the hypothesis that Wolbachia utilize the actin skeleton for efficient transmission and maintenance within Drosophila.

  18. Costs of Three Wolbachia Infections on the Survival of Aedes aegypti Larvae under Starvation Conditions

    PubMed Central

    Ross, Perran A.; Endersby, Nancy M.; Hoffmann, Ary A.

    2016-01-01

    The mosquito Aedes aegypti, the principal vector of dengue virus, has recently been infected experimentally with Wolbachia: intracellular bacteria that possess potential as dengue biological control agents. Wolbachia depend on their hosts for nutrients they are unable to synthesize themselves. Consequently, competition between Wolbachia and their host for resources could reduce host fitness under the competitive conditions commonly experienced by larvae of Ae. aegypti in the field, hampering the invasion of Wolbachia into natural mosquito populations. We assess the survival and development of Ae. aegypti larvae under starvation conditions when infected with each of three experimentally-generated Wolbachia strains: wMel, wMelPop and wAlbB, and compare their fitness to wild-type uninfected larvae. We find that all three Wolbachia infections reduce the survival of larvae relative to those that are uninfected, and the severity of the effect is concordant with previously characterized fitness costs to other life stages. We also investigate the ability of larvae to recover from extended food deprivation and find no effect of Wolbachia on this trait. Aedes aegypti larvae of all infection types were able to resume their development after one month of no food, pupate rapidly, emerge at a large size, and exhibit complete cytoplasmic incompatibility and maternal transmission. A lowered ability of Wolbachia-infected larvae to survive under starvation conditions will increase the threshold infection frequency required for Wolbachia to establish in highly competitive natural Ae. aegypti populations and will also reduce the speed of invasion. This study also provides insights into survival strategies of larvae when developing in stressful environments. PMID:26745630

  19. Absence of influential spreaders in rumor dynamics

    NASA Astrophysics Data System (ADS)

    Borge-Holthoefer, Javier; Moreno, Yamir

    2012-02-01

    Recent research [Kitsak, Gallos, Havlin, Liljeros, Muchnik, Stanley, and Makse, Nature Physics1745-247310.1038/nphys1746 6, 888 (2010)] has suggested that coreness, and not degree, constitutes a better topological descriptor to identify influential spreaders in complex networks. This hypothesis has been verified in the context of disease spreading. Here, we instead focus on rumor spreading models, which are more suited for social contagion and information propagation. To this end, we perform extensive computer simulations on top of several real-world networks and find opposite results. Namely, we show that the spreading capabilities of the nodes do not depend on their k-core index, which instead determines whether or not a given node prevents the diffusion of a rumor to a system-wide scale. Our findings are relevant both for sociological studies of contagious dynamics and for the design of efficient commercial viral processes.

  20. The mitochondrial heat shock protein 60 (HSP60) is up-regulated in Onchocerca volvulus after the depletion of Wolbachia.

    PubMed

    Pfarr, K M; Heider, U; Schmetz, C; Büttner, D W; Hoerauf, A

    2008-04-01

    Wolbachia, a genus of endosymbiotic bacteria of filarial worms, represent novel targets for anti-filarial therapy. The efficacy of compounds against Wolbachia has been evaluated using antiserum raised against the 60 kDa heat shock protein (HSP60) which binds specifically to this protein in both Wolbachia and mitochondria. It has been shown that Wolbachia stains (using such specific probes) stronger than the mitochondria in untreated Onchocerca volvulus, whereas after the depletion of Wolbachia (with drugs) staining of the mitochondria is increased. Herein, immunogold electron microscopy showed that specific anti-HSP60 serum specifically labelled Wolbachia and filarial mitochondria, and that both have distinct localization patterns, thus allowing them to be differentiated. Immunohistochemistry of O. volvulus showed that HSP60 staining is increased in the mitochondria after Wolbachia depletion in the hypodermis, epithelia, muscles, oocytes, embryos, and developing spermatozoa. This could have been the result of the antiserum preferentially binding to the Wolbachia when they are present or due to increased expression of the protein in the absence of the bacteria. To address this, mRNA levels of filarial hsp60 in O. volvulus were measured. After the depletion of Wolbachia, the transcription of hsp60 was significantly greater (7.7 fold) compared with untreated worms. We hypothesize that the increased expression of HSP60 in the absence of Wolbachia is due to a disruption of the homeostasis of the endosymbiosis.

  1. Helicopter crew/passenger vibration sensitivity -

    NASA Technical Reports Server (NTRS)

    Gabel, R.; Reed, D. A.

    1972-01-01

    Helicopter crew and passenger vibration sensitivity are presented. Pilot subjective ratings are established for discrete frequencies and the impact of combinations of harmonic frequencies is examined. A passenger long term comfort level and a short term limit are defined for discrete frequencies and compared with pilot ratings. The results show reasonable agreement between pilot and passenger. Subjective comfort levels obtained for mixed frequency environments clearly demonstrate the need for a multi-frequency criterion.

  2. New Insights into the Evolution of Wolbachia Infections in Filarial Nematodes Inferred from a Large Range of Screened Species

    PubMed Central

    Barbuto, Michela; Martin, Coralie; Lo, Nathan; Uni, Shigehiko; Landmann, Frederic; Baccei, Sara G.; Guerrero, Ricardo; de Souza Lima, Sueli; Bandi, Claudio; Wanji, Samuel; Diagne, Moustapha; Casiraghi, Maurizio

    2011-01-01

    Background Wolbachia are intriguing symbiotic endobacteria with a peculiar host range that includes arthropods and a single nematode family, the Onchocercidae encompassing agents of filariases. This raises the question of the origin of infection in filariae. Wolbachia infect the female germline and the hypodermis. Some evidences lead to the theory that Wolbachia act as mutualist and coevolved with filariae from one infection event: their removal sterilizes female filariae; all the specimens of a positive species are infected; Wolbachia are vertically inherited; a few species lost the symbiont. However, most data on Wolbachia and filaria relationships derive from studies on few species of Onchocercinae and Dirofilariinae, from mammals. Methodology/Principal Findings We investigated the Wolbachia distribution testing 35 filarial species, including 28 species and 7 genera and/or subgenera newly screened, using PCR, immunohistochemical staining, whole mount fluorescent analysis, and cocladogenesis analysis. (i) Among the newly screened Onchocercinae from mammals eight species harbour Wolbachia but for some of them, bacteria are absent in the hypodermis, or in variable density. (ii) Wolbachia are not detected in the pathological model Monanema martini and in 8, upon 9, species of Cercopithifilaria. (iii) Supergroup F Wolbachia is identified in two newly screened Mansonella species and in Cercopithifilaria japonica. (iv) Type F Wolbachia infect the intestinal cells and somatic female genital tract. (v) Among Oswaldofilariinae, Waltonellinae and Splendidofilariinae, from saurian, anuran and bird respectively, Wolbachia are not detected. Conclusions/Significance The absence of Wolbachia in 63% of onchocercids, notably in the ancestral Oswaldofilariinae estimated 140 mya old, the diverse tissues or specimens distribution, and a recent lateral transfer in supergroup F Wolbachia, modify the current view on the role and evolution of the endosymbiont and their hosts. Further

  3. US Advanced Freight and Passenger MAGLEV System

    NASA Technical Reports Server (NTRS)

    Morena, John J.; Danby, Gordon; Powell, James

    1996-01-01

    Japan and Germany will operate first generation Maglev passenger systems commercially shortly after 2000 A.D. The United States Maglev systems will require sophisticated freight and passenger carrying capability. The U.S. freight market is larger than passenger transport. A proposed advanced freight and passenger Maglev Project in Brevard County Florida is described. Present Maglev systems cost 30 million dollars or more per mile. Described is an advanced third generation Maglev system with technology improvements that will result in a cost of 10 million dollars per mile.

  4. 46 CFR 72.25-15 - Passenger accommodations for excursion boats, ferryboats, and passenger barges.

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    ... 46 Shipping 3 2014-10-01 2014-10-01 false Passenger accommodations for excursion boats, ferryboats, and passenger barges. 72.25-15 Section 72.25-15 Shipping COAST GUARD, DEPARTMENT OF HOMELAND SECURITY... accommodations for excursion boats, ferryboats, and passenger barges. (a) Except as specifically excluded by...

  5. 46 CFR 72.25-15 - Passenger accommodations for excursion boats, ferryboats, and passenger barges.

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ... 46 Shipping 3 2010-10-01 2010-10-01 false Passenger accommodations for excursion boats, ferryboats, and passenger barges. 72.25-15 Section 72.25-15 Shipping COAST GUARD, DEPARTMENT OF HOMELAND SECURITY... accommodations for excursion boats, ferryboats, and passenger barges. (a) Except as specifically excluded by...

  6. 46 CFR 72.25-15 - Passenger accommodations for excursion boats, ferryboats, and passenger barges.

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ... 46 Shipping 3 2013-10-01 2013-10-01 false Passenger accommodations for excursion boats, ferryboats, and passenger barges. 72.25-15 Section 72.25-15 Shipping COAST GUARD, DEPARTMENT OF HOMELAND SECURITY... accommodations for excursion boats, ferryboats, and passenger barges. (a) Except as specifically excluded by...

  7. 46 CFR 72.25-15 - Passenger accommodations for excursion boats, ferryboats, and passenger barges.

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ... 46 Shipping 3 2011-10-01 2011-10-01 false Passenger accommodations for excursion boats, ferryboats, and passenger barges. 72.25-15 Section 72.25-15 Shipping COAST GUARD, DEPARTMENT OF HOMELAND SECURITY... accommodations for excursion boats, ferryboats, and passenger barges. (a) Except as specifically excluded by...

  8. 46 CFR 72.25-15 - Passenger accommodations for excursion boats, ferryboats, and passenger barges.

    Code of Federal Regulations, 2012 CFR

    2012-10-01

    ... 46 Shipping 3 2012-10-01 2012-10-01 false Passenger accommodations for excursion boats, ferryboats, and passenger barges. 72.25-15 Section 72.25-15 Shipping COAST GUARD, DEPARTMENT OF HOMELAND SECURITY... accommodations for excursion boats, ferryboats, and passenger barges. (a) Except as specifically excluded by...

  9. Genomes of Candidatus Wolbachia bourtzisii wDacA and Candidatus Wolbachia pipientis wDacB from the Cochineal Insect Dactylopius coccus (Hemiptera: Dactylopiidae)

    PubMed Central

    Ramírez-Puebla, Shamayim T.; Ormeño-Orrillo, Ernesto; Vera-Ponce de León, Arturo; Lozano, Luis; Sanchez-Flores, Alejandro; Rosenblueth, Mónica; Martínez-Romero, Esperanza

    2016-01-01

    Dactylopius species, known as cochineal insects, are the source of the carminic acid dye used worldwide. The presence of two Wolbachia strains in Dactylopius coccus from Mexico was revealed by PCR amplification of wsp and sequencing of 16S rRNA genes. A metagenome analysis recovered the genome sequences of Candidatus Wolbachia bourtzisii wDacA (supergroup A) and Candidatus Wolbachia pipientis wDacB (supergroup B). Genome read coverage, as well as 16S rRNA clone sequencing, revealed that wDacB was more abundant than wDacA. The strains shared similar predicted metabolic capabilities that are common to Wolbachia, including riboflavin, ubiquinone, and heme biosynthesis, but lacked other vitamin and cofactor biosynthesis as well as glycolysis, the oxidative pentose phosphate pathway, and sugar uptake systems. A complete tricarboxylic acid cycle and gluconeogenesis were predicted as well as limited amino acid biosynthesis. Uptake and catabolism of proline were evidenced in Dactylopius Wolbachia strains. Both strains possessed WO-like phage regions and type I and type IV secretion systems. Several efflux systems found suggested the existence of metal toxicity within their host. Besides already described putative virulence factors like ankyrin domain proteins, VlrC homologs, and patatin-like proteins, putative novel virulence factors related to those found in intracellular pathogens like Legionella and Mycobacterium are highlighted for the first time in Wolbachia. Candidate genes identified in other Wolbachia that are likely involved in cytoplasmic incompatibility were found in wDacB but not in wDacA. PMID:27543297

  10. Genomes of Candidatus Wolbachia bourtzisii wDacA and Candidatus Wolbachia pipientis wDacB from the Cochineal Insect Dactylopius coccus (Hemiptera: Dactylopiidae).

    PubMed

    Ramírez-Puebla, Shamayim T; Ormeño-Orrillo, Ernesto; Vera-Ponce de León, Arturo; Lozano, Luis; Sanchez-Flores, Alejandro; Rosenblueth, Mónica; Martínez-Romero, Esperanza

    2016-10-13

    Dactylopius species, known as cochineal insects, are the source of the carminic acid dye used worldwide. The presence of two Wolbachia strains in Dactylopius coccus from Mexico was revealed by PCR amplification of wsp and sequencing of 16S rRNA genes. A metagenome analysis recovered the genome sequences of Candidatus Wolbachia bourtzisii wDacA (supergroup A) and Candidatus Wolbachia pipientis wDacB (supergroup B). Genome read coverage, as well as 16S rRNA clone sequencing, revealed that wDacB was more abundant than wDacA. The strains shared similar predicted metabolic capabilities that are common to Wolbachia, including riboflavin, ubiquinone, and heme biosynthesis, but lacked other vitamin and cofactor biosynthesis as well as glycolysis, the oxidative pentose phosphate pathway, and sugar uptake systems. A complete tricarboxylic acid cycle and gluconeogenesis were predicted as well as limited amino acid biosynthesis. Uptake and catabolism of proline were evidenced in Dactylopius Wolbachia strains. Both strains possessed WO-like phage regions and type I and type IV secretion systems. Several efflux systems found suggested the existence of metal toxicity within their host. Besides already described putative virulence factors like ankyrin domain proteins, VlrC homologs, and patatin-like proteins, putative novel virulence factors related to those found in intracellular pathogens like Legionella and Mycobacterium are highlighted for the first time in Wolbachia Candidate genes identified in other Wolbachia that are likely involved in cytoplasmic incompatibility were found in wDacB but not in wDacA.

  11. Analysis of bus passenger comfort perception based on passenger load factor and in-vehicle time.

    PubMed

    Shen, Xianghao; Feng, Shumin; Li, Zhenning; Hu, Baoyu

    2016-01-01

    Although bus comfort is a crucial indicator of service quality, existing studies tend to focus on passenger load and ignore in-vehicle time, which can also affect passengers' comfort perception. Therefore, by conducting surveys, this study examines passengers' comfort perception while accounting for both factors. Then, using the survey data, it performs a two-way analysis of variance and shows that both in-vehicle time and passenger load significantly affect passenger comfort. Then, a bus comfort model is proposed to evaluate comfort level, followed by a sensitivity analysis. The method introduced in this study has theoretical implications for bus operators attempting to improve bus service quality.

  12. A novel approach to eliminate Wolbachia infections in Nasonia vitripennis revealed different antibiotic resistance between two bacterial strains.

    PubMed

    Liu, Hai-Yang; Wang, Yan-Kun; Zhi, Cong-Cong; Xiao, Jin-Hua; Huang, Da-Wei

    2014-06-01

    Wolbachia are widespread in insects and can manipulate host reproduction. Nasonia vitripennis is a widely studied organism with a very high prevalence of Wolbachia infection. To study the effect of Wolbachia infection in Nasonia spp., it is important to obtain noninfected individuals by artificial methods. Current methods that employ sugar water-containing antibiotics can successfully eliminate Wolbachia from the parasitic wasps; however, treatment of at least three generations is required. Here, we describe a novel, feasible, and effective approach to eliminate Wolbachia from N. vitripennis by feeding fly pupae continuously offering antibiotics to Nasonia populations, which shortened the time to eliminate the pathogens to two generations. Additionally, the Wolbachia Uni and CauB strains have obviously different rifampicin-resistance abilities, which is a previously unknown phenomenon.

  13. Content Analysis of the 20 Most Influential Articles in "PIQ"

    ERIC Educational Resources Information Center

    Cho, Yonjoo; Park, Sunyoung

    2012-01-01

    The purpose of this study is to examine key research themes in human performance technology (HPT) through content analysis of the 20 most influential articles identified in Cho, Jo, Park, Kang, and Chen (2011). Three questions guiding this inquiry are: (1) What are the key themes of the 20 most influential articles in "PIQ", (2) What information…

  14. Evidence for Metabolic Provisioning by a Common Invertebrate Endosymbiont, Wolbachia pipientis, during Periods of Nutritional Stress

    PubMed Central

    Brownlie, Jeremy C.; Cass, Bodil N.; Riegler, Markus; Witsenburg, Joris J.; Iturbe-Ormaetxe, Iñaki; McGraw, Elizabeth A.; O'Neill, Scott L.

    2009-01-01

    Wolbachia are ubiquitous inherited endosymbionts of invertebrates that invade host populations by modifying host reproductive systems. However, some strains lack the ability to impose reproductive modification and yet are still capable of successfully invading host populations. To explain this paradox, theory predicts that such strains should provide a fitness benefit, but to date none has been detected. Recently completed genome sequences of different Wolbachia strains show that these bacteria may have the genetic machinery to influence iron utilization of hosts. Here we show that Wolbachia infection can confer a positive fecundity benefit for Drosophila melanogaster reared on iron-restricted or -overloaded diets. Furthermore, iron levels measured from field-collected flies indicated that nutritional conditions in the field were overall comparable to those of flies reared in the laboratory on restricted diets. These data suggest that Wolbachia may play a previously unrecognized role as nutritional mutualists in insects. PMID:19343208

  15. Wolbachia Do Not Induce Reactive Oxygen Species-Dependent Immune Pathway Activation in Aedes albopictus

    PubMed Central

    Molloy, Jennifer C.; Sinkins, Steven P.

    2015-01-01

    Aedes albopictus is a major vector of dengue (DENV) and chikungunya (CHIKV) viruses, causing millions of infections annually. It naturally carries, at high frequency, the intracellular inherited bacterial endosymbiont Wolbachia strains wAlbA and wAlbB; transinfection with the higher-density Wolbachia strain wMel from Drosophila melanogaster led to transmission blocking of both arboviruses. The hypothesis that reactive oxygen species (ROS)-induced immune activation plays a role in arbovirus inhibition in this species was examined. In contrast to previous observations in Ae. aegypti, elevation of ROS levels was not observed in either cell lines or mosquito lines carrying the wild-type Wolbachia or higher-density Drosophila Wolbachia strains. There was also no upregulation of genes controlling innate immune pathways or with antioxidant/ROS-producing functions. These data suggest that ROS-mediated immune activation is not an important component of the viral transmission-blocking phenotype in this species. PMID:26287231

  16. Closely related Wolbachia (Rickettsiales:Rickettsiaceae) recovered from different genera of Mexican Thelytokous figitidae (Hymenoptera)

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Thelytokous parasitoid strains are theoretically advantageous when utilized for biological control, as the absence of males should reduce production costs and potentially increase field efficacy. The maternally inherited intracellular bacterium, Wolbachia pipientis, is capable of inducing thelytokou...

  17. Tsetse-Wolbachia symbiosis: Comes of age and has great potential for pest and disease control

    PubMed Central

    Doudoumis, Vangelis; Alam, Uzma; Aksoy, Emre; Abd-Alla, Adly M.M.; Tsiamis, George; Brelsfoard, Corey; Aksoy, Serap; Bourtzis, Kostas

    2013-01-01

    Tsetse flies (Diptera: Glossinidae) are the sole vectors of African trypanosomes, the causative agent of sleeping sickness in human and nagana in animals. Like most eukaryotic organisms, Glossina species have established symbiotic associations with bacteria. Three main symbiotic bacteria have been found in tsetse flies: Wigglesworthia glossinidia, an obligate symbiotic bacterium, the secondary endosymbiont Sodalis glossinidius and the reproductive symbiont Wolbachia pipientis. In the present review, we discuss recent studies on the detection and characterization of Wolbachia infections in Glossina species, the horizontal transfer of Wolbachia genes to tsetse chromosomes, the ability of this symbiont to induce cytoplasmic incompatibility in Glossina morsitans morsitans and also how new environment-friendly tools for disease control could be developed by harnessing Wolbachia symbiosis. PMID:22835476

  18. Train Timetable Evaluation from the Viewpoints of Passengers by Microsimulation of Train Operation and Passenger Flow

    NASA Astrophysics Data System (ADS)

    Kunimatsu, Taketoshi; Hirai, Chikara; Tomii, Norio

    In order to evaluate train timetables from passengers' points of view, it is indispensable to estimate the disutilities of passengers. This can be done by estimating the movements of passengers and trains accurately. In particular, when there are many passengers, an interaction between the passengers and trains must be considered. To this end, we have developed a microsimulation system to simulate both train operation and passengers' train choice behavior. The system can simulate train choice behaviors of more than one million passengers as well as their positions in trains. It is possible to estimate the delays caused by congestion in trains as well. The system is based on models of different attitudes of the passengers with respect to the train choice behavior, with includes the choice of the earliest train, transfer avoidance, and congestion avoidance; a passengers' train choice behavior reflects his/her preferences. We applied this system to an actual railway line in a metropolitan area and evaluated two train schedules by calculating the generalized cost that reflects each passenger's disutility in his/her experience. Through the numerical experiments, we have confirmed that the proposed method is very useful for evaluating timetables from passengers' points of view.

  19. 14 CFR 91.1035 - Passenger awareness.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... 14 Aeronautics and Space 2 2012-01-01 2012-01-01 false Passenger awareness. 91.1035 Section 91.1035 Aeronautics and Space FEDERAL AVIATION ADMINISTRATION, DEPARTMENT OF TRANSPORTATION (CONTINUED... Operations Program Management § 91.1035 Passenger awareness. (a) Prior to each takeoff, the pilot in...

  20. 14 CFR 91.1035 - Passenger awareness.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 14 Aeronautics and Space 2 2010-01-01 2010-01-01 false Passenger awareness. 91.1035 Section 91.1035 Aeronautics and Space FEDERAL AVIATION ADMINISTRATION, DEPARTMENT OF TRANSPORTATION (CONTINUED... Operations Program Management § 91.1035 Passenger awareness. (a) Prior to each takeoff, the pilot in...

  1. 14 CFR 91.1035 - Passenger awareness.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... 14 Aeronautics and Space 2 2014-01-01 2014-01-01 false Passenger awareness. 91.1035 Section 91.1035 Aeronautics and Space FEDERAL AVIATION ADMINISTRATION, DEPARTMENT OF TRANSPORTATION (CONTINUED... Operations Program Management § 91.1035 Passenger awareness. (a) Prior to each takeoff, the pilot in...

  2. 14 CFR 91.1035 - Passenger awareness.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... 14 Aeronautics and Space 2 2011-01-01 2011-01-01 false Passenger awareness. 91.1035 Section 91.1035 Aeronautics and Space FEDERAL AVIATION ADMINISTRATION, DEPARTMENT OF TRANSPORTATION (CONTINUED... Operations Program Management § 91.1035 Passenger awareness. (a) Prior to each takeoff, the pilot in...

  3. 14 CFR 91.1035 - Passenger awareness.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... 14 Aeronautics and Space 2 2013-01-01 2013-01-01 false Passenger awareness. 91.1035 Section 91.1035 Aeronautics and Space FEDERAL AVIATION ADMINISTRATION, DEPARTMENT OF TRANSPORTATION (CONTINUED... Operations Program Management § 91.1035 Passenger awareness. (a) Prior to each takeoff, the pilot in...

  4. 14 CFR 91.517 - Passenger information.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... safety belt about him or her and keep it fastened while any “fasten seat belt” sign is lighted. (e) Each... smoking is prohibited and when safety belts must be fastened. The signs must be so constructed that the... belts and when smoking is prohibited. (c) If passenger information signs are installed, no passenger...

  5. 14 CFR 91.517 - Passenger information.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... safety belt about him or her and keep it fastened while any “fasten seat belt” sign is lighted. (e) Each... smoking is prohibited and when safety belts must be fastened. The signs must be so constructed that the... belts and when smoking is prohibited. (c) If passenger information signs are installed, no passenger...

  6. Wolbachia co-infection in a hybrid zone: discovery of horizontal gene transfers from two Wolbachia supergroups into an animal genome.

    PubMed

    Funkhouser-Jones, Lisa J; Sehnert, Stephanie R; Martínez-Rodríguez, Paloma; Toribio-Fernández, Raquel; Pita, Miguel; Bella, José L; Bordenstein, Seth R

    2015-01-01

    Hybrid zones and the consequences of hybridization have contributed greatly to our understanding of evolutionary processes. Hybrid zones also provide valuable insight into the dynamics of symbiosis since each subspecies or species brings its unique microbial symbionts, including germline bacteria such as Wolbachia, to the hybrid zone. Here, we investigate a natural hybrid zone of two subspecies of the meadow grasshopper Chorthippus parallelus in the Pyrenees Mountains. We set out to test whether co-infections of B and F Wolbachia in hybrid grasshoppers enabled horizontal transfer of phage WO, similar to the numerous examples of phage WO transfer between A and B Wolbachia co-infections. While we found no evidence for transfer between the divergent co-infections, we discovered horizontal transfer of at least three phage WO haplotypes to the grasshopper genome. Subsequent genome sequencing of uninfected grasshoppers uncovered the first evidence for two discrete Wolbachia supergroups (B and F) contributing at least 448 kb and 144 kb of DNA, respectively, into the host nuclear genome. Fluorescent in situ hybridization verified the presence of Wolbachia DNA in C. parallelus chromosomes and revealed that some inserts are subspecies-specific while others are present in both subspecies. We discuss our findings in light of symbiont dynamics in an animal hybrid zone.

  7. How Diverse Is the Genus Wolbachia? Multiple-Gene Sequencing Reveals a Putatively New Wolbachia Supergroup Recovered from Spider Mites (Acari: Tetranychidae)▿ †

    PubMed Central

    Ros, Vera I. D.; Fleming, Vicki M.; Feil, Edward J.; Breeuwer, Johannes A. J.

    2009-01-01

    At least 20% of all arthropods and some nematode species are infected with intracellular bacteria of the genus Wolbachia. This highly diverse genus has been subdivided into eight “supergroups” (A to H) on the basis of nucleotide sequence data. Here, we report the discovery of a new Wolbachia supergroup recovered from the spider mite species Bryobia species V (Acari: Tetranychidae), based on the sequences of three protein-coding genes (ftsZ, gltA, and groEL) and the 16S rRNA gene. Other tetranychid mites possess supergroup B Wolbachia strains. The discovery of another Wolbachia supergroup expands the known diversity of Wolbachia and emphasizes the high variability of the genus. Our data also clarify the existing supergroup structure and highlight the use of multiple gene sequences for robust phylogenetic analysis. In addition to previous reports of recombination between the arthropod-infecting supergroups A and B, we provide evidence for recombination between the nematode-infecting supergroups C and D. Robust delineation of supergroups is essential for understanding the origin and spread of this common reproductive parasite and for unraveling mechanisms of host adaptation and manipulation across a wide range of hosts. PMID:19098217

  8. Wolbachia co-infection in a hybrid zone: discovery of horizontal gene transfers from two Wolbachia supergroups into an animal genome

    PubMed Central

    Sehnert, Stephanie R.; Martínez-Rodríguez, Paloma; Toribio-Fernández, Raquel; Pita, Miguel; Bella, José L.; Bordenstein, Seth R.

    2015-01-01

    Hybrid zones and the consequences of hybridization have contributed greatly to our understanding of evolutionary processes. Hybrid zones also provide valuable insight into the dynamics of symbiosis since each subspecies or species brings its unique microbial symbionts, including germline bacteria such as Wolbachia, to the hybrid zone. Here, we investigate a natural hybrid zone of two subspecies of the meadow grasshopper Chorthippus parallelus in the Pyrenees Mountains. We set out to test whether co-infections of B and F Wolbachia in hybrid grasshoppers enabled horizontal transfer of phage WO, similar to the numerous examples of phage WO transfer between A and B Wolbachia co-infections. While we found no evidence for transfer between the divergent co-infections, we discovered horizontal transfer of at least three phage WO haplotypes to the grasshopper genome. Subsequent genome sequencing of uninfected grasshoppers uncovered the first evidence for two discrete Wolbachia supergroups (B and F) contributing at least 448 kb and 144 kb of DNA, respectively, into the host nuclear genome. Fluorescent in situ hybridization verified the presence of Wolbachia DNA in C. parallelus chromosomes and revealed that some inserts are subspecies-specific while others are present in both subspecies. We discuss our findings in light of symbiont dynamics in an animal hybrid zone. PMID:26664808

  9. Eukaryotic association module in phage WO genomes from Wolbachia

    PubMed Central

    Bordenstein, Sarah R.; Bordenstein, Seth R.

    2016-01-01

    Viruses are trifurcated into eukaryotic, archaeal and bacterial categories. This domain-specific ecology underscores why eukaryotic viruses typically co-opt eukaryotic genes and bacteriophages commonly harbour bacterial genes. However, the presence of bacteriophages in obligate intracellular bacteria of eukaryotes may promote DNA transfers between eukaryotes and bacteriophages. Here we report a metagenomic analysis of purified bacteriophage WO particles of Wolbachia and uncover a eukaryotic association module in the complete WO genome. It harbours predicted domains, such as the black widow latrotoxin C-terminal domain, that are uninterrupted in bacteriophage genomes, enriched with eukaryotic protease cleavage sites and combined with additional domains to forge one of the largest bacteriophage genes to date (14,256 bp). To the best of our knowledge, these eukaryotic-like domains have never before been reported in packaged bacteriophages and their phylogeny, distribution and sequence diversity imply lateral transfers between bacteriophage/prophage and animal genomes. Finally, the WO genome sequences and identification of attachment sites will potentially advance genetic manipulation of Wolbachia. PMID:27727237

  10. Detection and characterization of Wolbachia infections in laboratory and natural populations of different species of tsetse flies (genus Glossina)

    PubMed Central

    2012-01-01

    Background Wolbachia is a genus of endosymbiotic α-Proteobacteria infecting a wide range of arthropods and filarial nematodes. Wolbachia is able to induce reproductive abnormalities such as cytoplasmic incompatibility (CI), thelytokous parthenogenesis, feminization and male killing, thus affecting biology, ecology and evolution of its hosts. The bacterial group has prompted research regarding its potential for the control of agricultural and medical disease vectors, including Glossina spp., which transmits African trypanosomes, the causative agents of sleeping sickness in humans and nagana in animals. Results In the present study, we employed a Wolbachia specific 16S rRNA PCR assay to investigate the presence of Wolbachia in six different laboratory stocks as well as in natural populations of nine different Glossina species originating from 10 African countries. Wolbachia was prevalent in Glossina morsitans morsitans, G. morsitans centralis and G. austeni populations. It was also detected in G. brevipalpis, and, for the first time, in G. pallidipes and G. palpalis gambiensis. On the other hand, Wolbachia was not found in G. p. palpalis, G. fuscipes fuscipes and G. tachinoides. Wolbachia infections of different laboratory and natural populations of Glossina species were characterized using 16S rRNA, the wsp (Wolbachia Surface Protein) gene and MLST (Multi Locus Sequence Typing) gene markers. This analysis led to the detection of horizontal gene transfer events, in which Wobachia genes were inserted into the tsetse flies fly nuclear genome. Conclusions Wolbachia infections were detected in both laboratory and natural populations of several different Glossina species. The characterization of these Wolbachia strains promises to lead to a deeper insight in tsetse flies-Wolbachia interactions, which is essential for the development and use of Wolbachia-based biological control methods. PMID:22376025

  11. Influential input classification in probabilistic multimedia models

    SciTech Connect

    Maddalena, Randy L.; McKone, Thomas E.; Hsieh, Dennis P.H.; Geng, Shu

    1999-05-01

    Monte Carlo analysis is a statistical simulation method that is often used to assess and quantify the outcome variance in complex environmental fate and effects models. Total outcome variance of these models is a function of (1) the uncertainty and/or variability associated with each model input and (2) the sensitivity of the model outcome to changes in the inputs. To propagate variance through a model using Monte Carlo techniques, each variable must be assigned a probability distribution. The validity of these distributions directly influences the accuracy and reliability of the model outcome. To efficiently allocate resources for constructing distributions one should first identify the most influential set of variables in the model. Although existing sensitivity and uncertainty analysis methods can provide a relative ranking of the importance of model inputs, they fail to identify the minimum set of stochastic inputs necessary to sufficiently characterize the outcome variance. In this paper, we describe and demonstrate a novel sensitivity/uncertainty analysis method for assessing the importance of each variable in a multimedia environmental fate model. Our analyses show that for a given scenario, a relatively small number of input variables influence the central tendency of the model and an even smaller set determines the shape of the outcome distribution. For each input, the level of influence depends on the scenario under consideration. This information is useful for developing site specific models and improving our understanding of the processes that have the greatest influence on the variance in outcomes from multimedia models.

  12. Evolutionary Dynamics of wAu-Like Wolbachia Variants in Neotropical Drosophila spp.

    PubMed Central

    Miller, Wolfgang J.; Riegler, Markus

    2006-01-01

    Wolbachia bacteria are common intracellular symbionts of arthropods and have been extensively studied in Drosophila. Most research focuses on two Old Word hosts, Drosophila melanogaster and Drosophila simulans, and does not take into account that some of the Wolbachia associations in these species may have evolved only after their fast global expansion and after the exposure to Wolbachia of previously isolated habitats. Here we looked at Wolbachia of Neotropical Drosophila species. Seventy-one lines of 16 Neotropical Drosophila species sampled in different regions and at different time points were analyzed. Wolbachia is absent in lines of Drosophila willistoni collected before the 1970s, but more recent samples are infected with a strain designated wWil. Wolbachia is absent in all other species of the willistoni group. Polymorphic wWil-related strains were detected in some saltans group species, with D. septentriosaltans being coinfected with at least four variants. Based on wsp and ftsZ sequence data, wWil of D. willistoni is identical to wAu, a strain isolated from D. simulans, but can be discriminated when using a polymorphic minisatellite marker. In contrast to wAu, which infects both germ line and somatic tissues of D. simulans, wWil is found exclusively in the primordial germ line cells of D. willistoni embryos. We report on a pool of closely related Wolbachia strains in Neotropical Drosophila species as a potential source for the wAu strain in D. simulans. Possible evolutionary scenarios reconstructing the infection history of wAu-like Wolbachia in Neotropical Drosophila species and the Old World species D. simulans are discussed. PMID:16391124

  13. Wolbachia do not live by reproductive manipulation alone: infection polymorphism in Drosophila suzukii and D. subpulchrella

    PubMed Central

    Hamm, Christopher A.; Begun, David J.; Vo, Alexandre; Smith, Chris C. R.; Saelao, Perot; Shaver, Amanda O.; Jaenike, John; Turelli, Michael

    2014-01-01

    Drosophila suzukii recently invaded North America and Europe. Populations in Hawaii, California, New York and Nova Scotia are polymorphic for Wolbachia, typically with <20% infection frequency. The Wolbachia in D. suzukii, denoted wSuz, is closely related to wRi, the variant prevalent in continental populations of D. simulans. wSuz is also nearly identical to Wolbachia found in D. subpulchrella, plausibly D. suzukii's sister species. This suggests vertical Wolbachia transmission through cladogenesis (“cladogenic transmission”). The widespread occurrence of 7-20% infection frequencies indicates a stable polymorphism. wSuz is imperfectly maternally transmitted, with wild infected females producing on average 5-10% uninfected progeny. As expected from its low frequency, wSuz produces no cytoplasmic incompatibility (CI), i.e., no elevated embryo mortality when infected males mate with uninfected females, and no appreciable sex-ratio distortion. The persistence of wSuz despite imperfect maternal transmission suggests positive fitness effects. Assuming a balance between selection and imperfect transmission, we expect a fitness advantage on the order of 20%. Unexpectedly, Wolbachia-infected females produce fewer progeny than do uninfected females. We do not yet understand the maintenance of wSuz in D. suzukii. The absence of detectable CI in D. suzukii and D. subpulchrella makes it unlikely that CI-based mechanisms could be used to control this species without transinfection using novel Wolbachia. Contrary to their reputation as horizontally transmitted reproductive parasites, many Wolbachia infections are acquired through introgression or cladogenesis and many cause no appreciable reproductive manipulation. Such infections, likely to be mutualistic, may be central to understanding the pervasiveness of Wolbachia among arthropods. PMID:25156506

  14. Wolbachia-Induced Cytoplasmic Incompatibility Is Associated with Decreased Hira Expression in Male Drosophila

    PubMed Central

    Wang, Jia-Lin; Wang, Yu-Feng

    2011-01-01

    Background Wolbachia are obligate endosymbiotic bacteria that infect numerous species of arthropods and nematodes. Wolbachia can induce several reproductive phenotypes in their insect hosts including feminization, male-killing, parthenogenesis and cytoplasmic incompatibility (CI). CI is the most common phenotype and occurs when Wolbachia-infected males mate with uninfected females resulting in no or very low numbers of viable offspring. However, matings between males and females infected with the same strain of Wolbachia result in viable progeny. Despite substantial scientific effort, the molecular mechanisms underlying CI are currently unknown. Methodology/Principal Findings Gene expression studies were undertaken in Drosophila melanogaster and D. simulans which display differential levels of CI using quantitative RT-PCR. We show that Hira expression is correlated with the induction of CI and occurs in a sex-specific manner. Hira expression is significantly lower in males which induce strong CI when compared to males inducing no CI or Wolbachia-uninfected males. A reduction in Hira expression is also observed in 1-day-old males that induce stronger CI compared to 5-day-old males that induce weak or no CI. In addition, Hira mutated D. melanogaster males mated to uninfected females result in significantly decreased hatch rates comparing with uninfected crosses. Interestingly, wMel-infected females may rescue the hatch rates. An obvious CI phenotype with chromatin bridges are observed in the early embryo resulting from Hira mutant fertilization, which strongly mimics the defects associated with CI. Conclusions/Significance Our results suggest Wolbachia-induced CI in Drosophila occurs due to a reduction in Hira expression in Wolbachia-infected males leading to detrimental effects on sperm fertility resulting in embryo lethality. These results may help determine the underlying mechanism of CI and provide further insight in to the important role Hira plays in the

  15. Wolbachia do not live by reproductive manipulation alone: infection polymorphism in Drosophila suzukii and D. subpulchrella.

    PubMed

    Hamm, Christopher A; Begun, David J; Vo, Alexandre; Smith, Chris C R; Saelao, Perot; Shaver, Amanda O; Jaenike, John; Turelli, Michael

    2014-10-01

    Drosophila suzukii recently invaded North America and Europe. Populations in Hawaii, California, New York and Nova Scotia are polymorphic for Wolbachia, typically with <20% infection frequency. The Wolbachia in D. suzukii, denoted wSuz, is closely related to wRi, the variant prevalent in continental populations of D. simulans. wSuz is also nearly identical to Wolbachia found in D. subpulchrella, plausibly D. suzukii's sister species. This suggests vertical Wolbachia transmission through cladogenesis ('cladogenic transmission'). The widespread occurrence of 7-20% infection frequencies indicates a stable polymorphism. wSuz is imperfectly maternally transmitted, with wild infected females producing on average 5-10% uninfected progeny. As expected from its low frequency, wSuz produces no cytoplasmic incompatibility (CI), that is, no increased embryo mortality when infected males mate with uninfected females, and no appreciable sex-ratio distortion. The persistence of wSuz despite imperfect maternal transmission suggests positive fitness effects. Assuming a balance between selection and imperfect transmission, we expect a fitness advantage on the order of 20%. Unexpectedly, Wolbachia-infected females produce fewer progeny than do uninfected females. We do not yet understand the maintenance of wSuz in D. suzukii. The absence of detectable CI in D. suzukii and D. subpulchrella makes it unlikely that CI-based mechanisms could be used to control this species without transinfection using novel Wolbachia. Contrary to their reputation as horizontally transmitted reproductive parasites, many Wolbachia infections are acquired through introgression or cladogenesis and many cause no appreciable reproductive manipulation. Such infections, likely to be mutualistic, may be central to understanding the pervasiveness of Wolbachia among arthropods.

  16. Requirement of lipid II biosynthesis for cell division in cell wall-less Wolbachia, endobacteria of arthropods and filarial nematodes.

    PubMed

    Vollmer, Jennifer; Schiefer, Andrea; Schneider, Tanja; Jülicher, Karen; Johnston, Kelly L; Taylor, Mark J; Sahl, Hans-Georg; Hoerauf, Achim; Pfarr, Kenneth

    2013-04-01

    Obligate Wolbachia endobacteria have a reduced genome and retained genes are hypothesized to be crucial for survival. Although intracellular bacteria do not need a stress-bearing peptidoglycan cell wall, Wolbachia encode proteins necessary to synthesize the peptidoglycan precursor lipid II. The activity of the enzymes catalyzing the last two steps of this pathway was previously shown, and Wolbachia are sensitive to inhibition of lipid II synthesis. A puzzling characteristic of Wolbachia is the lack of genes for l-amino acid racemases essential for lipid II synthesis. Transcription analysis showed the expression of a possible alternative racemase metC, and recombinant Wolbachia MetC indeed had racemase activity that may substitute for the absent l-Ala racemase. However, enzymes needed to form mature peptidoglycan are absent and the function of Wolbachia lipid II is unknown. Inhibition of lipid II biosynthesis resulted in enlargement of Wolbachia cells and redistribution of Wolbachia peptidoglycan-associated lipoprotein, demonstrating that lipid II is required for coordinated cell division and may interact with the lipoprotein. We conclude that lipid II is essential for Wolbachia cell division and that this function is potentially conserved in the Gram-negative bacteria.

  17. Tropical tephritid fruit fly community with high incidence of shared Wolbachia strains as platform for horizontal transmission of endosymbionts.

    PubMed

    Morrow, J L; Frommer, M; Shearman, D C A; Riegler, M

    2014-12-01

    Wolbachia are endosymbiotic bacteria that infect 40-65% of arthropod species. They are primarily maternally inherited with occasional horizontal transmission for which limited direct ecological evidence exists. We detected Wolbachia in 8 out of 24 Australian tephritid species. Here, we have used multilocus sequence typing (MLST) to further characterize these Wolbachia strains, plus a novel quantitative polymerase chain reaction method for allele assignment in multiple infections. Based on five MLST loci and the Wolbachia surface protein gene (wsp), five Bactrocera and one Dacus species harboured two identical strains as double infections; furthermore, Bactrocera neohumeralis harboured both of these as single or double infections, and sibling species B. tryoni harboured one. Two Bactrocera species contained Wolbachia pseudogenes, potentially within the fruit fly genomes. A fruit fly parasitoid, Fopius arisanus shared identical alleles with two Wolbachia strains detected in one B. frauenfeldi individual. We report an unprecedented high incidence of four shared Wolbachia strains in eight host species from two trophic levels. This suggests frequent exposure to Wolbachia in this tropical tephritid community that shares host plant and parasitoid species, and also includes species that hybridize. Such insect communities may act as horizontal transmission platforms that contribute to the ubiquity of the otherwise maternally inherited Wolbachia.

  18. First record of Wolbachia in South American terrestrial isopods: Prevalence and diversity in two species of Balloniscus (Crustacea, Oniscidea)

    PubMed Central

    Almerão, Mauricio Pereira; Fagundes, Nelson Jurandi Rosa; de Araújo, Paula Beatriz; Verne, Sébastien; Grandjean, Frédéric; Bouchon, Didier; Araújo, Aldo Mellender

    2012-01-01

    Wolbachia are endosymbiotic bacteria that commonly infect arthropods, inducing certain phenotypes in their hosts. So far, no endemic South American species of terrestrial isopods have been investigated for Wolbachia infection. In this work, populations from two species of Balloniscus (B. sellowii and B. glaber) were studied through a diagnostic PCR assay. Fifteen new Wolbachia 16S rDNA sequences were detected. Wolbachia found in both species were generally specific to one population, and five populations hosted two different Wolbachia 16S rDNA sequences. Prevalence was higher in B. glaber than in B. sellowii, but uninfected populations could be found in both species. Wolbachia strains from B. sellowii had a higher genetic variation than those isolated from B. glaber. AMOVA analyses showed that most of the genetic variance was distributed among populations of each species rather than between species, and the phylogenetic analysis suggested that Wolbachia strains from Balloniscus cluster within Supergroup B, but do not form a single monophyletic clade, suggesting multiple infections for this group. Our results highlight the importance of studying Wolbachia prevalence and genetic diversity in Neotropical species and suggest that South American arthropods may harbor a great number of diverse strains, providing an interesting model to investigate the evolution of Wolbachia and its hosts. PMID:23413179

  19. Tunga penetrans: molecular identification of Wolbachia endobacteria and their recognition by antibodies against proteins of endobacteria from filarial parasites.

    PubMed

    Fischer, Peter; Schmetz, Christel; Bandi, Claudio; Bonow, Insa; Mand, Sabine; Fischer, Kerstin; Büttner, Dietrich W

    2002-01-01

    In search of Wolbachia in human parasites, Wolbachia were identified in the sand flea Tunga penetrans. PCR and DNA sequencing of the bacterial 16S rDNA, the ftsZ cell division protein, the Wolbachia surface protein (wsp) and the Wolbachia aspartate aminotransferase genes revealed a high similarity to the respective sequences of endosymbionts of filarial nematodes. Using these sequences a phylogenetic tree was generated, that indicates a close relationship between Wolbachia from T. penetrans and from filarial parasites, but possibly as a member of a new supergroup. Ultrastructural studies showed that Wolbachia are abundant in the ovaries of neosomic fleas, whereas other, smaller and morphologically distinct, bacteria were observed in the lumen of the intestine. Wolbachia were labeled by immunohistology and immunogold electron microscopy using polyclonal antibodies against wsp of Drosophila, of the filarial parasite Dirofilaria immitis, or against hsp 60 from Yersinia enterocolitica. These results show that as in filariasis, humans with tungiasis are exposed to Wolbachia. Furthermore, antisera raised against proteins of Wolbachia from arthropods or from filarial parasites can be immunologically cross-reactive.

  20. Evidence for selective sweeps by Wolbachia infections: phylogeny of Altica leaf beetles and their reproductive parasites.

    PubMed

    Jäckel, Regina; Mora, Diana; Dobler, Susanne

    2013-08-01

    Infections with maternally inherited Wolbachia bacteria may have dramatic influences on reproductive traits and speciation patterns of their hosts. We here show that in the beetle genus Altica, infection has influenced phylogenetic patterns of the host's mtDNA and different strains led to repeated selective sweeps. By comparing a COI/II-based phylogeny of the hosts with a phylogeny of the bacteria based on ftsZ, we show that cospeciation is rare and restricted to few recently diverged species. While in general each species apparently harbours a single Wolbachia strain, Altica lythri presents a strikingly different pattern: in the polyphyletic species, three highly divergent mtDNA haplotypes (2.1-4.6% p-distance) are coupled with three different Wolbachia strains (wLytA1, wLytA2 and wLytB). These haplotypes and Wolbachia strains are widely distributed and mostly found in sympatry. A phylogeny based on microsatellite data supports the monophyly of A. lythri. The discrepancy between mtDNA and nuclear phylogeny may best be explained by interspecific hybridization that led to introgression of mtDNA coupled with a different Wolbachia strain. Selective sweeps apparently drove the introgressed haplotypes to widespread distribution. As for effects of Wolbachia on reproduction, infection with wLytA1 appears to be correlated with a substantial sex ratio distortion, which was most prominent in A. lythri.

  1. Genomic evidence for plant-parasitic nematodes as the earliest Wolbachia hosts

    PubMed Central

    Brown, Amanda M. V.; Wasala, Sulochana K.; Howe, Dana K.; Peetz, Amy B.; Zasada, Inga A.; Denver, Dee R.

    2016-01-01

    Wolbachia, one of the most widespread endosymbionts, is a target for biological control of mosquito-borne diseases (malaria and dengue virus), and antibiotic elimination of infectious filarial nematodes. We sequenced and analyzed the genome of a new Wolbachia strain (wPpe) in the plant-parasitic nematode Pratylenchus penetrans. Phylogenomic analyses placed wPpe as the earliest diverging Wolbachia, suggesting two evolutionary invasions into nematodes. The next branches comprised strains in sap-feeding insects, suggesting Wolbachia may have first evolved as a nutritional mutualist. Genome size, protein content, %GC, and repetitive DNA allied wPpe with mutualistic Wolbachia, whereas gene repertoire analyses placed it between parasite (A, B) and mutualist (C, D, F) groups. Conservation of iron metabolism genes across Wolbachia suggests iron homeostasis as a potential factor in its success. This study enhances our understanding of this globally pandemic endosymbiont, highlighting genetic patterns associated with host changes. Combined with future work on this strain, these genomic data could help provide potential new targets for plant-parasitic nematode control. PMID:27734894

  2. Dynamics of Wolbachia pipientis Gene Expression Across the Drosophila melanogaster Life Cycle

    PubMed Central

    Gutzwiller, Florence; Carmo, Catarina R.; Miller, Danny E.; Rice, Danny W.; Newton, Irene L. G.; Hawley, R. Scott; Teixeira, Luis; Bergman, Casey M.

    2015-01-01

    Symbiotic interactions between microbes and their multicellular hosts have manifold biological consequences. To better understand how bacteria maintain symbiotic associations with animal hosts, we analyzed genome-wide gene expression for the endosymbiotic α-proteobacteria Wolbachia pipientis across the entire life cycle of Drosophila melanogaster. We found that the majority of Wolbachia genes are expressed stably across the D. melanogaster life cycle, but that 7.8% of Wolbachia genes exhibit robust stage- or sex-specific expression differences when studied in the whole-organism context. Differentially-expressed Wolbachia genes are typically up-regulated after Drosophila embryogenesis and include many bacterial membrane, secretion system, and ankyrin repeat-containing proteins. Sex-biased genes are often organized as small operons of uncharacterized genes and are mainly up-regulated in adult Drosophila males in an age-dependent manner. We also systematically investigated expression levels of previously-reported candidate genes thought to be involved in host-microbe interaction, including those in the WO-A and WO-B prophages and in the Octomom region, which has been implicated in regulating bacterial titer and pathogenicity. Our work provides comprehensive insight into the developmental dynamics of gene expression for a widespread endosymbiont in its natural host context, and shows that public gene expression data harbor rich resources to probe the functional basis of the Wolbachia-Drosophila symbiosis and annotate the transcriptional outputs of the Wolbachia genome. PMID:26497146

  3. Tetracycline treatment targeting Wolbachia affects expression of an array of proteins in Brugia malayi parasite.

    PubMed

    Dangi, Anil; Vedi, Satish; Nag, Jeetendra Kumar; Paithankar, Sameer; Singh, Mahendra Pratap; Kar, Santosh Kumar; Dube, Anuradha; Misra-Bhattacharya, Shailja

    2009-09-01

    Wolbachia is an intracellular endosymbiont of Brugia malayi parasite whose presence is essential for the survival of the parasite. Treatment of B. malayi-infected jirds with tetracycline eliminates Wolbachia, which affects parasite survival and fitness. In the present study we have tried to identify parasite proteins that are affected when Wolbachia is targeted by tetracycline. For this Wolbachia depleted parasites (B. malayi) were obtained by tetracycline treatment of infected Mongolian jirds (Meriones unguiculatus) and their protein profile after 2-DE separation was compared with that of untreated parasites harboring Wolbachia. Approximately 100 protein spots could be visualized followed by CBB staining of 2-D gel and included for comparative analysis. Of these, 54 showed differential expressions, while two new protein spots emerged (of 90.3 and 64.4 kDa). These proteins were subjected to further analysis by MALDI-TOF for their identification using Brugia coding sequence database composed of both genomic and EST sequences. Our study unravels two crucial findings: (i) the parasite or Wolbachia proteins, which disappeared/down-regulated appear be essential for parasite survival and may be used as drug targets and (ii) tetracycline treatment interferes with the regulatory machinery vital for parasites cellular integrity and defense and thus could possibly be a molecular mechanism for the killing of filarial parasite. This is the first proteomic study substantiating the wolbachial genome integrity with its nematode host and providing functional genomic data of human lymphatic filarial parasite B. malayi.

  4. Multiple Horizontal Transfers of Bacteriophage WO and Host Wolbachia in Fig Wasps in a Closed Community

    PubMed Central

    Wang, Ningxin; Jia, Sisi; Xu, Heng; Liu, Yong; Huang, Dawei

    2016-01-01

    Wolbachia-bacteriophage WO is a good model system for studying interactions between bacteria and viruses. Previous surveys of insect hosts have been conducted via sampling from open or semi-open communities; however, no studies have reported the infection patterns of phage WO of insects living in a closed community. Figs and fig wasps form a peculiar closed community in which the Ficus tree provides a compact syconium habitat for a variety of fig wasp. Therefore, in this study, we performed a thorough survey of Wolbachia and bacteriophage WO infection patterns in a total of 1406 individuals from 23 fig wasps species living on three different fig tree species. The infection rates of Wolbachia and phage WO were 82.6% (19/23) and 39.1% (9/23), respectively. Additionally, phage WO from fig wasps showed strong insect host specificity based on orf7 sequences from fig wasps and 21 other insect species. Probably due to the physical barrier of fig syconium, most phage WO from fig wasps form a specific clade. Phylogenetic analysis showed the absence of congruence between WO and host Wolbachia, WO and insect host, as well as Wolbachia and fig wasps, suggesting that both Wolbachia and phage WO exchanged frequently and independently within the closed syconium. Thus, the infection pattern of bacteriophage WO from fig wasps appeared quite different from that in other insects living outside, although the effect and the transfer routes of phage WO are unclear, which need to be investigated in the future. PMID:26913026

  5. Detection of the Wolbachia Protein WPIP0282 in Mosquito Spermathecae: Implications for Cytoplasmic Incompatibility

    PubMed Central

    Beckmann, John F.; Fallon, Ann M.

    2013-01-01

    Cytoplasmic incompatibility (CI) is a conditional sterility induced by the bacterium Wolbachia pipientis that infects reproductive tissues in many arthropods. Although CI provides a potential tool to control insect vectors of arthropod-borne diseases, the molecular basis for CI induction is unknown. We hypothesized that a Wolbachia-encoded, CI-inducing factor would be enriched in sperm recovered from spermathecae of female mosquitoes. Using SDS-PAGE and mass spectrometry, we detected peptides from the 56 kDa hypothetical protein, encoded by wPip_0282, associated with sperm transferred to females by Wolbachia infected males. We also detected peptides from the same protein in Wolbachia infected ovaries. Homologs of wPip_0282 and the co-transcribed downstream gene, wPip_0283, occur as multiple divergent copies in genomes of CI-inducing strains of Wolbachia. The operon is located in a genomic context that includes mobile genetic elements. The absence of wPip_0282 and wPip_0283 homologs from genomes of Wolbachia in filarial nematodes, as well as other members of the Rickettsiales, suggests a role as a candidate CI effector. PMID:23856508

  6. Broader prevalence of Wolbachia in insects including potential human disease vectors.

    PubMed

    de Oliveira, C D; Gonçalves, D S; Baton, L A; Shimabukuro, P H F; Carvalho, F D; Moreira, L A

    2015-06-01

    Wolbachia are intracellular, maternally transmitted bacteria considered the most abundant endosymbionts found in arthropods. They reproductively manipulate their host in order to increase their chances of being transmitted to the offspring, and currently are being used as a tool to control vector-borne diseases. Studies on distribution of Wolbachia among its arthropod hosts are important both for better understanding why this bacterium is so common, as well as for its potential use as a biological control agent. Here, we studied the incidence of Wolbachia in a broad range of insect species, collected from different regions of Brazil, using three genetic markers (16S rRNA, wsp and ftsZ), which varied in terms of their sensitivity to detect this bacterium. The overall incidence of Wolbachia among species belonging to 58 families and 14 orders was 61.9%. The most common positive insect orders were Coleoptera, Diptera, Hemiptera and Hymenoptera, with Diptera and Hemiptera having the highest numbers of Wolbachia-positive families. They included potential human disease vectors whose infection status has never been reported before. Our study further shows the importance of using quantitative polymerase chain reaction for high-throughput and sensitive Wolbachia screening.

  7. Wolbachia density and virulence attenuation after transfer into a novel host.

    PubMed

    McGraw, E A; Merritt, D J; Droller, J N; O'Neill, S L

    2002-03-05

    The factors that control replication rate of the intracellular bacterium Wolbachia pipientis in its insect hosts are unknown and difficult to explore, given the complex interaction of symbiont and host genotypes. Using a strain of Wolbachia that is known to over-replicate and shorten the lifespan of its Drosophila melanogaster host, we have tracked the evolution of replication control in both somatic and reproductive tissues in a novel host/Wolbachia association. After transinfection (the transfer of a Wolbachia strain into a different species) of the over-replicating Wolbachia popcorn strain from D. melanogaster to Drosophila simulans, we demonstrated that initial high densities in the ovaries were in excess of what was required for perfect maternal transmission, and were likely causing reductions in reproductive fitness. Both densities and fitness costs associated with ovary infection rapidly declined in the generations after transinfection. The early death effect in D. simulans attenuated only slightly and was comparable to that induced in D. melanogaster. This study reveals a strong host involvement in Wolbachia replication rates, the independence of density control responses in different tissues, and the strength of natural selection acting on reproductive fitness.

  8. The Hematopoietic Organ: A Cornerstone for Wolbachia Propagation Between and Within Hosts

    PubMed Central

    Braquart-Varnier, Christine; Raimond, Maryline; Mappa, Gaëtan; Chevalier, Frédéric D.; Le Clec’h, Winka; Sicard, Mathieu

    2015-01-01

    Wolbachia is an intracellular α-proteobacterium which is transmitted vertically from mother to offspring but also frequently switches horizontally from one host to another. Our hypothesis is based on the role of immune cells and the organs that produce them, the hematopoietic organs (HOs), as primordial niches for the propagation of Wolbachia via hemocytes both (i) within hosts: to initiate and maintain the systemic infection and (ii) between hosts: to promote both vertical and horizontal transmission of Wolbachia. Therefore, we review some fundamental ideas underlying this hypothesis and go further with new empirical data that lead to a first close-up analysis of the potential role of HOs in Wolbachia propagation. The monitoring of the first steps of Wolbachia infection in horizontally infected host organs by transmission electron microscopy and qPCR suggests that (i) HOs are colonized early and extensively as soon as they are in contact with Wolbachia which find in these cells a favorable niche to multiply and (ii) infected HOs which expel hemocytes all lifelong can generate and maintain a systemic infection that could contribute to increase both vertical and horizontal propagation of these symbionts. PMID:26733969

  9. The intracellular bacterium Wolbachia uses parasitoid wasps as phoretic vectors for efficient horizontal transmission.

    PubMed

    Ahmed, Muhammad Z; Li, Shao-Jian; Xue, Xia; Yin, Xiang-Jie; Ren, Shun-Xiang; Jiggins, Francis M; Greeff, Jaco M; Qiu, Bao-Li

    2015-02-01

    Facultative bacterial endosymbionts are associated with many arthropods and are primarily transmitted vertically from mother to offspring. However, phylogenetic affiliations suggest that horizontal transmission must also occur. Such horizontal transfer can have important biological and agricultural consequences when endosymbionts increase host fitness. So far horizontal transmission is considered rare and has been difficult to document. Here, we use fluorescence in situ hybridization (FISH) and multi locus sequence typing (MLST) to reveal a potentially common pathway of horizontal transmission of endosymbionts via parasitoids of insects. We illustrate that the mouthparts and ovipositors of an aphelinid parasitoid become contaminated with Wolbachia when this wasp feeds on or probes Wolbachia-infected Bemisia tabaci AsiaII7, and non-lethal probing of uninfected B. tabaci AsiaII7 nymphs by parasitoids carrying Wolbachia resulted in newly and stably infected B. tabaci matrilines. After they were exposed to infected whitefly, the parasitoids were able to transmit Wolbachia efficiently for the following 48 h. Whitefly infected with Wolbachia by parasitoids had increased survival and reduced development times. Overall, our study provides evidence for the horizontal transmission of Wolbachia between insect hosts by parasitic wasps, and the enhanced survival and reproductive abilities of insect hosts may adversely affect biological control programs.

  10. Multiple Wolbachia determinants control the evolution of cytoplasmic incompatibilities in Culex pipiens mosquito populations.

    PubMed

    Atyame, Celestine M; Duron, Olivier; Tortosa, Pablo; Pasteur, Nicole; Fort, Philippe; Weill, Mylene

    2011-01-01

    Wolbachia are maternally inherited endosymbionts that can invade arthropod populations through manipulation of their reproduction. In mosquitoes, Wolbachia induce embryonic death, known as cytoplasmic incompatibility (CI), whenever infected males mate with females either uninfected or infected with an incompatible strain. Although genetic determinants of CI are unknown, a functional model involving the so-called mod and resc factors has been proposed. Natural populations of Culex pipiens mosquito display a complex CI relationship pattern associated with the highest Wolbachia (wPip) genetic polymorphism reported so far. We show here that C. pipiens populations from La Réunion, a geographically isolated island in the southwest of the Indian Ocean, are infected with genetically closely related wPip strains. Crossing experiments reveal that these Wolbachia are all mutually compatible. However, crosses with genetically more distant wPip strains indicate that Wolbachia strains from La Réunion belong to at least five distinct incompatibility groups (or crossing types). These incompatibility properties which are strictly independent from the nuclear background, formally establish that in C. pipiens, CI is controlled by several Wolbachia mod/resc factors.

  11. Influence of two Wolbachia strains on population structure of East African Drosophila simulans.

    PubMed Central

    Dean, Matthew D; Ballard, Kirrie J; Glass, Anne; Ballard, J William O

    2003-01-01

    Drosophila simulans is hypothesized to have originated in continental East Africa or Madagascar. In this study, we investigated evolutionary forces operating on mitochondrial DNA (mtDNA) in populations of D. simulans from Zimbabwe, Malawi, Tanzania, and Kenya. Variation in mtDNA may be affected by positive selection, background selection, demographic history, and/or any maternally inherited factor such as the bacterial symbiont Wolbachia. In East Africa, the wRi and wMa Wolbachia strains associate with the siII or siIII mitochondrial haplogroups, respectively. To ask how polymorphism relates to Wolbachia infection status, we sequenced 1776 bp of mitochondrial DNA and 1029 bp of the X-linked per locus from 79 lines. The two southern populations were infected with wRi and exhibited significantly reduced mtDNA variation, while Wolbachia-uninfected siII flies from Tanzania and Kenya showed high levels of mtDNA polymorphism. These are the first known populations of D. simulans that do not exhibit reduced mtDNA variation. We observed no mitochondrial variation in the siIII haplogroup regardless of Wolbachia infection status, suggesting positive or background selection. These populations offer a unique opportunity to monitor evolutionary dynamics in ancestral populations that harbor multiple strains of Wolbachia. PMID:14704179

  12. Multiple Horizontal Transfers of Bacteriophage WO and Host Wolbachia in Fig Wasps in a Closed Community.

    PubMed

    Wang, Ningxin; Jia, Sisi; Xu, Heng; Liu, Yong; Huang, Dawei

    2016-01-01

    Wolbachia-bacteriophage WO is a good model system for studying interactions between bacteria and viruses. Previous surveys of insect hosts have been conducted via sampling from open or semi-open communities; however, no studies have reported the infection patterns of phage WO of insects living in a closed community. Figs and fig wasps form a peculiar closed community in which the Ficus tree provides a compact syconium habitat for a variety of fig wasp. Therefore, in this study, we performed a thorough survey of Wolbachia and bacteriophage WO infection patterns in a total of 1406 individuals from 23 fig wasps species living on three different fig tree species. The infection rates of Wolbachia and phage WO were 82.6% (19/23) and 39.1% (9/23), respectively. Additionally, phage WO from fig wasps showed strong insect host specificity based on orf7 sequences from fig wasps and 21 other insect species. Probably due to the physical barrier of fig syconium, most phage WO from fig wasps form a specific clade. Phylogenetic analysis showed the absence of congruence between WO and host Wolbachia, WO and insect host, as well as Wolbachia and fig wasps, suggesting that both Wolbachia and phage WO exchanged frequently and independently within the closed syconium. Thus, the infection pattern of bacteriophage WO from fig wasps appeared quite different from that in other insects living outside, although the effect and the transfer routes of phage WO are unclear, which need to be investigated in the future.

  13. Dynamics of Wolbachia pipientis Gene Expression Across the Drosophila melanogaster Life Cycle.

    PubMed

    Gutzwiller, Florence; Carmo, Catarina R; Miller, Danny E; Rice, Danny W; Newton, Irene L G; Hawley, R Scott; Teixeira, Luis; Bergman, Casey M

    2015-10-23

    Symbiotic interactions between microbes and their multicellular hosts have manifold biological consequences. To better understand how bacteria maintain symbiotic associations with animal hosts, we analyzed genome-wide gene expression for the endosymbiotic α-proteobacteria Wolbachia pipientis across the entire life cycle of Drosophila melanogaster. We found that the majority of Wolbachia genes are expressed stably across the D. melanogaster life cycle, but that 7.8% of Wolbachia genes exhibit robust stage- or sex-specific expression differences when studied in the whole-organism context. Differentially-expressed Wolbachia genes are typically up-regulated after Drosophila embryogenesis and include many bacterial membrane, secretion system, and ankyrin repeat-containing proteins. Sex-biased genes are often organized as small operons of uncharacterized genes and are mainly up-regulated in adult Drosophila males in an age-dependent manner. We also systematically investigated expression levels of previously-reported candidate genes thought to be involved in host-microbe interaction, including those in the WO-A and WO-B prophages and in the Octomom region, which has been implicated in regulating bacterial titer and pathogenicity. Our work provides comprehensive insight into the developmental dynamics of gene expression for a widespread endosymbiont in its natural host context, and shows that public gene expression data harbor rich resources to probe the functional basis of the Wolbachia-Drosophila symbiosis and annotate the transcriptional outputs of the Wolbachia genome.

  14. Wolbachia small noncoding RNAs and their role in cross-kingdom communications.

    PubMed

    Mayoral, Jaime G; Hussain, Mazhar; Joubert, D Albert; Iturbe-Ormaetxe, Iñaki; O'Neill, Scott L; Asgari, Sassan

    2014-12-30

    In prokaryotes, small noncoding RNAs (snRNAs) of 50-500 nt are produced that are important in bacterial virulence and response to environmental stimuli. Here, we identified and characterized snRNAs from the endosymbiotic bacteria, Wolbachia, which are widespread in invertebrates and cause reproductive manipulations. Most importantly, some strains of Wolbachia inhibit replication of several vector-borne pathogens in insects. We demonstrate that two abundant snRNAs, WsnRNA-46 and WsnRNA-49, are expressed in Wolbachia from noncoding RNA transcripts that contain precursors with stem-loop structures. WsnRNAs were detected in Aedes aegypti mosquitoes infected with the wMelPop-CLA strain of Wolbachia and in Drosophila melanogaster and Drosophila simulans infected with wMelPop and wAu strains, respectively, indicating that the WsnRNAs are conserved across species and strains. In addition, we show that the WsnRNAs may potentially regulate host genes and Wolbachia genes. Our findings provide evidence for the production of functional snRNAs by Wolbachia that play roles in cross-kingdom communication between the endosymbiont and the host.

  15. Proteomic profiling of a robust Wolbachia infection in an Aedes albopictus mosquito cell line

    PubMed Central

    Baldridge, Gerald D; Baldridge, Abigail S; Witthuhn, Bruce A; Higgins, LeeAnn; Markowski, Todd W; Fallon, Ann M

    2014-01-01

    Wolbachia pipientis a widespread vertically transmitted intracellular bacterium, provides a tool for insect control through manipulation of host-microbe interactions. We report proteomic characterization of wStr, a Wolbachia strain associated with a strong cytoplasmic incompatibility phenotype in its native host, Laodelphax striatellus. In the Aedes albopictus C/wStr1 mosquito cell line, wStr maintains a robust, persistent infection. MS/MS analyses of gel bands revealed a protein “footprint” dominated by Wolbachia-encoded chaperones, stress response and cell membrane proteins, including the surface antigen WspA, a peptidoglycan-associated lipoprotein and a 73 kDa outer membrane protein. Functional classifications and estimated abundance levels of 790 identified proteins suggested that expression, stabilization and secretion of proteins predominate over bacterial genome replication and cell division. High relative abundances of cysteine desulfurase, serine/glycine hydroxymethyl transferase, and components of the α-ketoglutarate dehydrogenase complex in conjunction with above average abundances of glutamate dehydrogenase and proline utilization protein A support Wolbachia genome-based predictions for amino acid metabolism as a primary energy source. wStr expresses 15 Vir proteins of a Type IV secretion system and its transcriptional regulator. Proteomic characterization of a robust insect-associated Wolbachia strain provides baseline information that will inform further development of in vitro protocols for Wolbachia manipulation. PMID:25155417

  16. An endosymbiotic bacterium in a plant-parasitic nematode: member of a new Wolbachia supergroup.

    PubMed

    Haegeman, Annelies; Vanholme, Bartel; Jacob, Joachim; Vandekerckhove, Tom T M; Claeys, Myriam; Borgonie, Gaetan; Gheysen, Godelieve

    2009-07-15

    Wolbachia is an endosymbiotic bacterium widely present in arthropods and animal-parasitic nematodes. Despite previous efforts, it has never been identified in plant-parasitic nematodes. Random sequencing of genes expressed by the burrowing nematode Radopholus similis resulted in several sequences with similarity to Wolbachia genes. The presence of a Wolbachia-like endosymbiont in this plant-parasitic nematode was investigated using both morphological and molecular approaches. Transmission electronmicroscopy, fluorescent immunolocalisation and staining with DAPI confirmed the presence of the endosymbiont within the reproductive tract of female adults. 16S rDNA, ftsZ and groEL gene sequences showed that the endosymbiont of R. similis is distantly related to the known Wolbachia supergroups. Finally, based on our initial success in finding sequences of this endosymbiont by screening an expressed sequence tag (EST) dataset, all nematode ESTs were mined for Wolbachia-like sequences. Although the retained sequences belonged to six different nematode species, R. similis was the only plant-parasitic nematode with traces of Wolbachia. Based on our phylogenetic study and the current literature we designate the endosymbiont of R. similis to a new supergroup (supergroup I) rather than considering it as a new species. Although its role remains unknown, the endosymbiont was found in all individuals tested, pointing towards an essential function of the bacteria.

  17. Evolution of cooperation in a heterogeneous population with influential individuals

    NASA Astrophysics Data System (ADS)

    Zhuang, Qian; Wang, Dong; Fan, Ying; Di, Zengru

    2012-02-01

    Influential individuals are introduced and integrated with the public goods game (PGG) to investigate their influence on the emergence and evolution of cooperation. In the model, some influential individuals whose behaviors can be controlled by us are introduced into a homogeneous population on a square lattice. The influential individuals can play three kinds of roles: I. exemplar, II. supervisor with the power to punish defectors, and III. supervisor with the power to reward cooperative co-players. It is found that the existence of influential individuals who play Role I turns out to be detrimental to cooperation and that the larger the number of influential individuals is, the more difficult it is for cooperation to be maintained. For those playing supervisory roles, both punishment and reward are found to be effective ways for the influential individuals to promote and stabilize cooperative behavior. By comparing the critical costs and the mean payoffs for a low multiplication factor under the role of punishment and the role of reward, it is found that reward is a more effective intervention measure than punishment for influential individuals seeking to improve cooperation and that reward leads to a higher mean payoff.

  18. Effects of Doxycycline on gene expression in Wolbachia and Brugia malayi adult female worms in vivo

    PubMed Central

    2012-01-01

    Background Most filarial nematodes contain Wolbachia symbionts. The purpose of this study was to examine the effects of doxycycline on gene expression in Wolbachia and adult female Brugia malayi. Methods Brugia malayi infected gerbils were treated with doxycycline for 6-weeks. This treatment largely cleared Wolbachia and arrested worm reproduction. RNA recovered from treated and control female worms was labeled by random priming and hybridized to the Version 2- filarial microarray to obtain expression profiles. Results and discussion Results showed significant changes in expression for 200 Wolbachia (29% of Wolbachia genes with expression signals in untreated worms) and 546 B. malayi array elements after treatment. These elements correspond to known genes and also to novel genes with unknown biological functions. Most differentially expressed Wolbachia genes were down-regulated after treatment (98.5%). In contrast, doxycycline had a mixed effect on B. malayi gene expression with many more genes being significantly up-regulated after treatment (85% of differentially expressed genes). Genes and processes involved in reproduction (gender-regulated genes, collagen, amino acid metabolism, ribosomal processes, and cytoskeleton) were down-regulated after doxycycline while up-regulated genes and pathways suggest adaptations for survival in response to stress (energy metabolism, electron transport, anti-oxidants, nutrient transport, bacterial signaling pathways, and immune evasion). Conclusions Doxycycline reduced Wolbachia and significantly decreased bacterial gene expression. Wolbachia ribosomes are believed to be the primary biological target for doxycycline in filarial worms. B. malayi genes essential for reproduction, growth and development were also down-regulated; these changes are consistent with doxycycline effects on embryo development and reproduction. On the other hand, many B. malayi genes involved in energy production, electron-transport, metabolism, anti

  19. Wolbachia infections in Anopheles gambiae cells: transcriptomic characterization of a novel host-symbiont interaction.

    PubMed

    Hughes, Grant L; Ren, Xiaoxia; Ramirez, Jose L; Sakamoto, Joyce M; Bailey, Jason A; Jedlicka, Anne E; Rasgon, Jason L

    2011-02-01

    The endosymbiotic bacterium Wolbachia is being investigated as a potential control agent in several important vector insect species. Recent studies have shown that Wolbachia can protect the insect host against a wide variety of pathogens, resulting in reduced transmission of parasites and viruses. It has been proposed that compromised vector competence of Wolbachia-infected insects is due to up-regulation of the host innate immune system or metabolic competition. Anopheles mosquitoes, which transmit human malaria parasites, have never been found to harbor Wolbachia in nature. While transient somatic infections can be established in Anopheles, no stable artificially-transinfected Anopheles line has been developed despite numerous attempts. However, cultured Anopheles cells can be stably infected with multiple Wolbachia strains such as wAlbB from Aedes albopictus, wRi from Drosophila simulans and wMelPop from Drosophila melanogaster. Infected cell lines provide an amenable system to investigate Wolbachia-Anopheles interactions in the absence of an infected mosquito strain. We used Affymetrix GeneChip microarrays to investigate the effect of wAlbB and wRi infection on the transcriptome of cultured Anopheles Sua5B cells, and for a subset of genes used quantitative PCR to validate results in somatically-infected Anopheles mosquitoes. Wolbachia infection had a dramatic strain-specific effect on gene expression in this cell line, with almost 700 genes in total regulated representing a diverse array of functional classes. Very strikingly, infection resulted in a significant down-regulation of many immune, stress and detoxification-related transcripts. This is in stark contrast to the induction of immune genes observed in other insect hosts. We also identified genes that may be potentially involved in Wolbachia-induced reproductive and pathogenic phenotypes. Somatically-infected mosquitoes had similar responses to cultured cells. The data show that Wolbachia has a profound

  20. 46 CFR 46.05-25 - New passenger vessel.

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ... 46 Shipping 2 2010-10-01 2010-10-01 false New passenger vessel. 46.05-25 Section 46.05-25 Shipping COAST GUARD, DEPARTMENT OF HOMELAND SECURITY (CONTINUED) LOAD LINES SUBDIVISION LOAD LINES FOR PASSENGER VESSELS Definitions Used in This Part § 46.05-25 New passenger vessel. A new passenger vessel is a...

  1. 49 CFR 523.5 - Non-passenger automobile.

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ... 49 Transportation 6 2010-10-01 2010-10-01 false Non-passenger automobile. 523.5 Section 523.5... ADMINISTRATION, DEPARTMENT OF TRANSPORTATION VEHICLE CLASSIFICATION § 523.5 Non-passenger automobile. A non-passenger automobile means an automobile that is not a passenger automobile or a work truck and...

  2. 46 CFR 46.05-25 - New passenger vessel.

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ... 46 Shipping 2 2011-10-01 2011-10-01 false New passenger vessel. 46.05-25 Section 46.05-25 Shipping COAST GUARD, DEPARTMENT OF HOMELAND SECURITY (CONTINUED) LOAD LINES SUBDIVISION LOAD LINES FOR PASSENGER VESSELS Definitions Used in This Part § 46.05-25 New passenger vessel. A new passenger vessel is a...

  3. 49 CFR 177.870 - Regulations for passenger carrying vehicles.

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    ... PUBLIC HIGHWAY Regulations Applying to Hazardous Material on Motor Vehicles Carrying Passengers for Hire § 177.870 Regulations for passenger carrying vehicles. (a) Vehicles transporting passengers and property... apply to vehicles transporting passengers and property. (b) No Class 1 (explosive) materials or...

  4. 46 CFR 176.910 - Passenger Ship Safety Certificate.

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ... 46 Shipping 7 2013-10-01 2013-10-01 false Passenger Ship Safety Certificate. 176.910 Section 176..., as Amended (SOLAS) § 176.910 Passenger Ship Safety Certificate. (a) A vessel, which carries more than 12 passengers on an international voyage must have a valid SOLAS Passenger Ship Safety...

  5. 46 CFR 176.910 - Passenger Ship Safety Certificate.

    Code of Federal Regulations, 2012 CFR

    2012-10-01

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  6. 46 CFR 176.910 - Passenger Ship Safety Certificate.

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ... 46 Shipping 7 2010-10-01 2010-10-01 false Passenger Ship Safety Certificate. 176.910 Section 176..., as Amended (SOLAS) § 176.910 Passenger Ship Safety Certificate. (a) A vessel, which carries more than 12 passengers on an international voyage must have a valid SOLAS Passenger Ship Safety...

  7. 46 CFR 176.910 - Passenger Ship Safety Certificate.

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ... 46 Shipping 7 2011-10-01 2011-10-01 false Passenger Ship Safety Certificate. 176.910 Section 176..., as Amended (SOLAS) § 176.910 Passenger Ship Safety Certificate. (a) A vessel, which carries more than 12 passengers on an international voyage must have a valid SOLAS Passenger Ship Safety...

  8. 46 CFR 176.910 - Passenger Ship Safety Certificate.

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    ... 46 Shipping 7 2014-10-01 2014-10-01 false Passenger Ship Safety Certificate. 176.910 Section 176..., as Amended (SOLAS) § 176.910 Passenger Ship Safety Certificate. (a) A vessel, which carries more than 12 passengers on an international voyage must have a valid SOLAS Passenger Ship Safety...

  9. 49 CFR 523.5 - Non-passenger automobile.

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    ... 49 Transportation 6 2014-10-01 2014-10-01 false Non-passenger automobile. 523.5 Section 523.5... ADMINISTRATION, DEPARTMENT OF TRANSPORTATION VEHICLE CLASSIFICATION § 523.5 Non-passenger automobile. A non-passenger automobile means an automobile that is not a passenger automobile or a work truck and...

  10. 49 CFR 523.5 - Non-passenger automobile.

    Code of Federal Regulations, 2012 CFR

    2012-10-01

    ... 49 Transportation 6 2012-10-01 2012-10-01 false Non-passenger automobile. 523.5 Section 523.5... ADMINISTRATION, DEPARTMENT OF TRANSPORTATION VEHICLE CLASSIFICATION § 523.5 Non-passenger automobile. A non-passenger automobile means an automobile that is not a passenger automobile or a work truck and...

  11. 49 CFR 523.5 - Non-passenger automobile.

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ... 49 Transportation 6 2011-10-01 2011-10-01 false Non-passenger automobile. 523.5 Section 523.5... ADMINISTRATION, DEPARTMENT OF TRANSPORTATION VEHICLE CLASSIFICATION § 523.5 Non-passenger automobile. A non-passenger automobile means an automobile that is not a passenger automobile or a work truck and...

  12. 49 CFR 523.5 - Non-passenger automobile.

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ... 49 Transportation 6 2013-10-01 2013-10-01 false Non-passenger automobile. 523.5 Section 523.5... ADMINISTRATION, DEPARTMENT OF TRANSPORTATION VEHICLE CLASSIFICATION § 523.5 Non-passenger automobile. A non-passenger automobile means an automobile that is not a passenger automobile or a work truck and...

  13. 19 CFR 122.88 - Aircraft carrying domestic (stopover) passengers.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 19 Customs Duties 1 2011-04-01 2011-04-01 false Aircraft carrying domestic (stopover) passengers... Passengers § 122.88 Aircraft carrying domestic (stopover) passengers. Airlines that commingle domestic... continuing on another aircraft to a second U.S. destination) with international passengers who are...

  14. 19 CFR 122.88 - Aircraft carrying domestic (stopover) passengers.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

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  15. 19 CFR 122.88 - Aircraft carrying domestic (stopover) passengers.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

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  16. 19 CFR 122.88 - Aircraft carrying domestic (stopover) passengers.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

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  17. 49 CFR 223.15 - Requirements for existing passenger cars.

    Code of Federal Regulations, 2013 CFR

    2013-10-01

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  18. 49 CFR 223.15 - Requirements for existing passenger cars.

    Code of Federal Regulations, 2011 CFR

    2011-10-01

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  19. 49 CFR 223.15 - Requirements for existing passenger cars.

    Code of Federal Regulations, 2012 CFR

    2012-10-01

    ... 49 Transportation 4 2012-10-01 2012-10-01 false Requirements for existing passenger cars. 223.15... ADMINISTRATION, DEPARTMENT OF TRANSPORTATION SAFETY GLAZING STANDARDS-LOCOMOTIVES, PASSENGER CARS AND CABOOSES Specific Requirements § 223.15 Requirements for existing passenger cars. (a) Passenger cars built...

  20. 49 CFR 223.15 - Requirements for existing passenger cars.

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ... 49 Transportation 4 2010-10-01 2010-10-01 false Requirements for existing passenger cars. 223.15... ADMINISTRATION, DEPARTMENT OF TRANSPORTATION SAFETY GLAZING STANDARDS-LOCOMOTIVES, PASSENGER CARS AND CABOOSES Specific Requirements § 223.15 Requirements for existing passenger cars. (a) Passenger cars built...

  1. 49 CFR 223.15 - Requirements for existing passenger cars.

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    ... 49 Transportation 4 2014-10-01 2014-10-01 false Requirements for existing passenger cars. 223.15... ADMINISTRATION, DEPARTMENT OF TRANSPORTATION SAFETY GLAZING STANDARDS-LOCOMOTIVES, PASSENGER CARS AND CABOOSES Specific Requirements § 223.15 Requirements for existing passenger cars. (a) Passenger cars built...

  2. Sex and stripping: The key to the intimate relationship between Wolbachia and host?

    PubMed

    Negri, Ilaria; Pellecchia, Marco; Grève, Pierre; Daffonchio, Daniele; Bandi, Claudio; Alma, Alberto

    2010-03-01

    Wolbachia pipientis is known to infect only arthropods and nematodes (mainly filarial worms). A unique feature shared by the two Phyla is the ability to replace the exoskeleton, a process known as ecdysis. This shared characteristic is thought to reflect a common ancestry. Arthropod moulting is induced by the steroid hormone 20-hydroxyecdysone (20E) and a role for ecdysteroids in nematode ecdysis has also been suggested. Removing Wolbachia from filarial worms impairs the host's development. From analyses of the genome of Wolbachia harbored by the filarial nematode Brugia malayi and that of its host, the bacterium may provide a source of heme, an essential component of cytochrome P450's that are necessary for steroid hormone biosynthetic pathways.In arthropods, Wolbachia is a reproductive manipulator, inducing various phenotypic effects that may be due to differences in host physiology, in particular, endocrine-related processes governing development and reproduction. Insect steroids have well-defined roles in the coordination of multiple developmental processes, and in adults they control important aspects of reproduction, including ovarian development, oogenesis, sexual behavior, and in some taxa vitellogenin biosynthesis.According to some authors ecdysteroids may also act as sex hormones. In insects sex differentiation is generally thought to be a strictly genetic process, in which each cell decides its own sexual fate based on its sex chromosome constitution, but, surprisingly, recent data demonstrate that in Drosophila sex determination is not cell-autonomous, as it happens in mammals. Thus the presence of signals coordinating the development of a gender-specific phenotype cannot be excluded.This could explain why Wolbachia interferes with insect reproduction; and also could explain why Wolbachia interferes with insect development.Thus, is "sex (=reproduction) and stripping (=ecdysis)" the key to the intimate relationship between Wolbachia and its host?

  3. Widespread infection and diverse infection patterns of Wolbachia in Chinese aphids.

    PubMed

    Wang, Zhe; Su, Xiao-Min; Wen, Juan; Jiang, Li-Yun; Qiao, Ge-Xia

    2014-06-01

    Wolbachia are intracellular symbionts that infect a wide range of arthropods and filarial nematodes. Aphids are engaged in diverse and complex relationships with their endosymbionts. Four supergroups (A, B, M and N) of Wolbachia were previously detected in aphids and supergroups M and N were only found in aphids. In this study, we detected and described Wolbachia infections in natural populations of aphids in China. Three supergroups (A, B and M) were found in the examined aphid species. Supergroup M was preponderant, whereas supergroups A and B were only detected in certain species. Supergroup N was not found in this study. There were four infection patterns of Wolbachia in aphids, namely, infection with supergroup M alone, co-infection with supergroups A and M, co-infection with supergroups B and M, and co-infection with supergroups A, B and M. The pattern of infection only with supergroup M was universal and was found in all evaluated subfamilies. Only two subfamilies, Aphidinae and Lachninae, manifested to present all four infection patterns. Three patterns were observed in Calaphidinae (M, A&M, B&M) and Eriosomatinae (M, B&M, A&B&M). Two patterns were observed in the Anoeciinae (M, A&M) and Greenideinae (M, B&M), and only one pattern (M) was observed in the remaining families and/or subfamilies of Aphidoidea. These results indicated that Wolbachia infections in Chinese aphids are widespread. Phylogenetic analyses suggest that Wolbachia supergroup M spread rapidly and recently among all host species of aphids in China. Reasons for this spread and its mechanisms are discussed along with the possible effects of Wolbachia on their aphid hosts.

  4. Diet-Induced Nutritional Stress and Pathogen Interference in Wolbachia-Infected Aedes aegypti

    PubMed Central

    Caragata, Eric Pearce; Rezende, Fernanda Oliveira; Simões, Taynãna César; Moreira, Luciano Andrade

    2016-01-01

    The pathogen interference phenotype greatly restricts infection with dengue virus (DENV) and other pathogens in Wolbachia-infected Aedes aegypti, and is a vital component of Wolbachia-based mosquito control. Critically, the phenotype’s causal mechanism is complex and poorly understood, with recent evidence suggesting that the cause may be species specific. To better understand this important phenotype, we investigated the role of diet-induced nutritional stress on interference against DENV and the avian malarial parasite Plasmodium gallinaceum in Wolbachia-infected Ae. aegypti, and on physiological processes linked to the phenotype. Wolbachia-infected mosquitoes were fed one of four different concentrations of sucrose, and then challenged with either P. gallinaceum or DENV. Interference against P. gallinaceum was significantly weakened by the change in diet however there was no effect on DENV interference. Immune gene expression and H2O2 levels have previously been linked to pathogen interference. These traits were assayed for mosquitoes on each diet using RT-qPCR and the Amplex Red Hydrogen Peroxide/Peroxidase Assay Kit, and it was observed that the change in diet did not significantly affect immune expression, but low carbohydrate levels led to a loss of ROS induction in Wolbachia-infected mosquitoes. Our data suggest that host nutrition may not influence DENV interference for Wolbachia-infected mosquitoes, but Plasmodium interference may be linked to both nutrition and oxidative stress. This pathogen-specific response to nutritional change highlights the complex nature of interactions between Wolbachia and pathogens in mosquitoes. PMID:27893736

  5. The distribution of Wolbachia in Cubitermes (Termitidae, Termitinae) castes and colonies: a modelling approach.

    PubMed

    Roy, Virginie; Girondot, Marc; Harry, Myriam

    2015-01-01

    Wolbachia are endosymbiotic bacteria of arthropods and nematodes that are able to manipulate host reproduction. Although vertically transmitted via the cytoplasm in eggs, horizontal transmission of Wolbachia among and within arthropod species has been shown to be common. Eusocial insects represent interesting models for studying Wolbachia transmission due to colonial organization and close interaction between nestmates. Here we conducted a detailed screening of Wolbachia infection for 15 colonies of the very common soil-feeding termites Cubitermes spp. affinis subarquatus (Termitidae, Termitinae) that consist of four distinct phylogenetic species in the Lopé forest Reserve, Gabon. Infection tests showed that 50% of the individuals were Wolbachia positive (N = 555) with 90% of reproductives and 48% of offspring infected. White soldiers, which are transitional stages preceding mature soldiers, had a significantly higher mean infection rate (74%) than the other castes and stages (63%, 33% and 39% for larvae, workers and mature soldiers, respectively). We used a maximum likelihood method and Akaike's Information Criterion in order to explain the non-expected high rate of Wolbachia infection in white soldiers. The best model included a species effect for the stochastic loss of Wolbachia and a caste effect for the rate of gain. After fitting, the best model selected was for a species-specific rate of loss with a null rate of new gain for larvae, workers and soldiers and a probability of 0.72 whatever the species, that a white soldier becomes newly contaminated during that stage. The mean expected infection rate in white soldiers without a new gain was estimated to 17% instead of the 74% observed. Here we discuss the possible explanations to the high infection rate observed in white soldiers.

  6. The Distribution of Wolbachia in Cubitermes (Termitidae, Termitinae) Castes and Colonies: A Modelling Approach

    PubMed Central

    Roy, Virginie; Girondot, Marc; Harry, Myriam

    2015-01-01

    Wolbachia are endosymbiotic bacteria of arthropods and nematodes that are able to manipulate host reproduction. Although vertically transmitted via the cytoplasm in eggs, horizontal transmission of Wolbachia among and within arthropod species has been shown to be common. Eusocial insects represent interesting models for studying Wolbachia transmission due to colonial organization and close interaction between nestmates. Here we conducted a detailed screening of Wolbachia infection for 15 colonies of the very common soil-feeding termites Cubitermes spp. affinis subarquatus (Termitidae, Termitinae) that consist of four distinct phylogenetic species in the Lopé forest Reserve, Gabon. Infection tests showed that 50% of the individuals were Wolbachia positive (N = 555) with 90% of reproductives and 48% of offspring infected. White soldiers, which are transitional stages preceding mature soldiers, had a significantly higher mean infection rate (74%) than the other castes and stages (63%, 33% and 39% for larvae, workers and mature soldiers, respectively). We used a maximum likelihood method and Akaike’s Information Criterion in order to explain the non-expected high rate of Wolbachia infection in white soldiers. The best model included a species effect for the stochastic loss of Wolbachia and a caste effect for the rate of gain. After fitting, the best model selected was for a species-specific rate of loss with a null rate of new gain for larvae, workers and soldiers and a probability of 0.72 whatever the species, that a white soldier becomes newly contaminated during that stage. The mean expected infection rate in white soldiers without a new gain was estimated to 17% instead of the 74% observed. Here we discuss the possible explanations to the high infection rate observed in white soldiers. PMID:25671520

  7. Identifying influential and susceptible members of social networks.

    PubMed

    Aral, Sinan; Walker, Dylan

    2012-07-20

    Identifying social influence in networks is critical to understanding how behaviors spread. We present a method that uses in vivo randomized experimentation to identify influence and susceptibility in networks while avoiding the biases inherent in traditional estimates of social contagion. Estimation in a representative sample of 1.3 million Facebook users showed that younger users are more susceptible to influence than older users, men are more influential than women, women influence men more than they influence other women, and married individuals are the least susceptible to influence in the decision to adopt the product offered. Analysis of influence and susceptibility together with network structure revealed that influential individuals are less susceptible to influence than noninfluential individuals and that they cluster in the network while susceptible individuals do not, which suggests that influential people with influential friends may be instrumental in the spread of this product in the network.

  8. 49 CFR 238.307 - Periodic mechanical inspection of passenger cars and unpowered vehicles used in passenger trains.

    Code of Federal Regulations, 2012 CFR

    2012-10-01

    ... 49 Transportation 4 2012-10-01 2012-10-01 false Periodic mechanical inspection of passenger cars... Passenger Equipment § 238.307 Periodic mechanical inspection of passenger cars and unpowered vehicles used in passenger trains. (a) General. (1) Railroads shall conduct periodic mechanical inspections of...

  9. 49 CFR 238.307 - Periodic mechanical inspection of passenger cars and unpowered vehicles used in passenger trains.

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ... 49 Transportation 4 2011-10-01 2011-10-01 false Periodic mechanical inspection of passenger cars... Passenger Equipment § 238.307 Periodic mechanical inspection of passenger cars and unpowered vehicles used in passenger trains. (a) General. (1) Railroads shall conduct periodic mechanical inspections of...

  10. 49 CFR 238.307 - Periodic mechanical inspection of passenger cars and unpowered vehicles used in passenger trains.

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ... 49 Transportation 4 2010-10-01 2010-10-01 false Periodic mechanical inspection of passenger cars... Passenger Equipment § 238.307 Periodic mechanical inspection of passenger cars and unpowered vehicles used in passenger trains. (a) General. (1) Railroads shall conduct periodic mechanical inspections of...

  11. 14 CFR 136.7 - Passenger briefings.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... AND OPERATORS FOR COMPENSATION OR HIRE: CERTIFICATION AND OPERATIONS COMMERCIAL AIR TOURS AND NATIONAL PARKS AIR TOUR MANAGEMENT National Air Tour Safety Standards § 136.7 Passenger briefings. (a)...

  12. 78 FR 49248 - Passenger Vessels Accessibility Guidelines

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-08-13

    ... TRANSPORTATION BARRIERS COMPLIANCE BOARD 36 CFR Part 1196 RIN 3014-AA11 Passenger Vessels Accessibility Guidelines AGENCY: Architectural and Transportation Barriers Compliance Board. ACTION: Notice of proposed rulemaking; extension of comment period. SUMMARY: The Architectural and Transportation Barriers...

  13. 46 CFR 176.113 - Passengers permitted.

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ... equipment or line handling gear, in the way of sail booms, running rigging, or paddle wheels, or along... of sail booms or running rigging; (v) Spaces below deck that are unsuitable for passengers or...

  14. 46 CFR 176.113 - Passengers permitted.

    Code of Federal Regulations, 2012 CFR

    2012-10-01

    ... equipment or line handling gear, in the way of sail booms, running rigging, or paddle wheels, or along... of sail booms or running rigging; (v) Spaces below deck that are unsuitable for passengers or...

  15. 46 CFR 176.113 - Passengers permitted.

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    ... equipment or line handling gear, in the way of sail booms, running rigging, or paddle wheels, or along... of sail booms or running rigging; (v) Spaces below deck that are unsuitable for passengers or...

  16. 46 CFR 176.113 - Passengers permitted.

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ... or line handling gear, in the way of sail booms, running rigging, or paddle wheels, or along pulpits... of sail booms or running rigging; (v) Spaces below deck that are unsuitable for passengers or...

  17. Locating influential nodes via dynamics-sensitive centrality.

    PubMed

    Liu, Jian-Guo; Lin, Jian-Hong; Guo, Qiang; Zhou, Tao

    2016-02-24

    With great theoretical and practical significance, locating influential nodes of complex networks is a promising issue. In this paper, we present a dynamics-sensitive (DS) centrality by integrating topological features and dynamical properties. The DS centrality can be directly applied in locating influential spreaders. According to the empirical results on four real networks for both susceptible-infected-recovered (SIR) and susceptible-infected (SI) spreading models, the DS centrality is more accurate than degree, k-shell index and eigenvector centrality.

  18. Incomplete removal of Wolbachia with tetracycline has two-edged reproductive effects in the thelytokous wasp Encarsia formosa (Hymenoptera: Aphelinidae)

    PubMed Central

    Wang, Xiao-Xiang; Qi, Lan-Da; Jiang, Rui; Du, Yu-Zhou; Li, Yuan-Xi

    2017-01-01

    Wolbachia pipientis are intracellular endosymbionts that induce parthenogenesis in the parasitoid Encarsia formosa. Previous studies that focused on effects of Wolbachia on the wasp usually used tetracycline to remove Wolbachia without concern for the joint influences of tetracycline and Wolbachia. Here we treated the wasps (F0 lines) with tetracycline to produce offspring (F1 lines) which were not fed tetracycline to avoid antibiotic influence. The quantitative data and fluorescence in situ hybridization showed that Wolbachia titers were reduced but not totally removed. The Wolbachia that infected the male offspring were unpredictably detected. Low dose tetracycline enhanced the fertility of 2-day-old F0 wasps after 24 h of treatment; however, compared with controls, the oocyte load of 3- to 6-day-old tetracycline-treated wasps decreased day by day, and tetracycline reduced the longevity of the wasps. The fecundity of controls was significantly higher than that of the treated F1-10 and F1-20 generations. Gene expression of vitellogenin reflected the same trend as that of wasp fecundities in both F0 and F1 lines. Moreover, female offspring proportions of F0 and F1 lines were related to the titer of infected Wolbachia, demonstrating that Wolbachia titer affected the sex determination of E. formosa. PMID:28266601

  19. Rapid and non-destructive detection and identification two strains of Wolbachia in Aedes aegypti by near-infrared spectroscopy

    Technology Transfer Automated Retrieval System (TEKTRAN)

    We investigated the potential of using near-infrared spectroscopy (NIRS) to detect the presence of Wolbachia pipientis (wMel) in male and female laboratory-reared Aedes aegypti mosquitoes. The release of Wolbachia transinfected mosquitoes is likely to form a key component of disease control strategi...

  20. Variable Infection Frequency and High Diversity of Multiple Strains of Wolbachia pipientis in Perkinsiella Planthoppers ▿ †

    PubMed Central

    Hughes, G. L.; Allsopp, P. G.; Brumbley, S. M.; Woolfit, M.; McGraw, E. A.; O'Neill, S. L.

    2011-01-01

    This survey of Wolbachia infections in populations of the planthoppers Perkinsiella saccharicida and Perkinsiella vitiensis revealed variable frequencies, low-titer infections, and high phylogenetic diversities of strains. These observations add to the growing realization that Wolbachia infections may be extremely common within invertebrates and yet occur infrequently within populations and at low titer within individuals. PMID:21278277

  1. Effects of parasitic sex-ratio distorters on host genetic structure in the Armadillidium vulgare-Wolbachia association.

    PubMed

    Verne, S; Johnson, M; Bouchon, D; Grandjean, F

    2012-02-01

    In the pill bug Armadillidium vulgare (Crustacea, Oniscidea), Wolbachia facilitates its spread through vertical transmission via the eggs by inducing feminization of genetic males. The spread of feminizing Wolbachia within and across populations is therefore expected to influence mitochondrial DNA (mtDNA) genetic structure by hitchhiking. To test this hypothesis, we analysed nuclear and mtDNA genetic structure, and Wolbachia prevalence in 13 populations of the pill bug host. Wolbachia prevalence (ranging from 0% to 100% of sampled females) was highly variable among populations. All three Wolbachia strains previously observed in A. vulgare were present (wVulC, wVulM and wVulP) with wVulC being the most prevalent (nine of 13 populations). The host showed a genetic structure on five microsatellite loci that is compatible with isolation by distance. The strong genetic structure observed on host mtDNA was correlated with Wolbachia prevalence: three mitotypes were in strong linkage disequilibrium with the three strains of Wolbachia. Neutrality tests showed that the mtDNA polymorphism is not neutral, and we thus suggest that this unusual pattern of mtDNA polymorphism found in A. vulgare was due to Wolbachia.

  2. A short, high-temperature treatment of host larvae to analyze Wolbachia-host interactions in the moth Ostrinia scapulalis.

    PubMed

    Sugimoto, Takafumi N; Kayukawa, Takumi; Matsuo, Takashi; Tsuchida, Tsutomu; Ishikawa, Yukio

    2015-10-01

    Maternally inherited endosymbiotic bacteria of the genus Wolbachia cause various reproductive alterations in their hosts. Wolbachia induces male-specific death during embryonic and larval stages in the moth Ostrinia scapulalis. To investigate how the density of Wolbachia affects their performance in the host, we attempted to reduce its density using a short, high-temperature treatment of the host at the larval stage. Individuals cured of infection as well as sexual mosaics, which harbor Wolbachia, were obtained by this method in the next generation. The sex of uninfected offspring was exclusively male, similar to that of the offspring of larvae treated with antibiotics. A strong correlation was found between Wolbachia density in female moths and the sex ratio of their progeny. These results suggest that a short, high-temperature treatment at the larval stage reduced the density of Wolbachia in the adult stage, and, hence, inhibited interference with the host's development in the next generation. Since the direct effects of the heat treatment on Wolbachia were transient, this method may be useful for specifying the critical time for interference by Wolbachia in host development.

  3. Wolbachia effects on host fitness and the influence of male aging on cytoplasmic incompatibility in Aedes polynesiensis (Diptera: Culicidae).

    PubMed

    Brelsfoard, C L; Dobson, S L

    2011-09-01

    The endosymbiotic bacteria Wolbachia manipulate host reproduction by inducing a form of sterility known as cytoplasmic incompatibility (CI), promoting the invasion of infection into natural host populations. CI has received attention for use in applied strategies to control insect vectors of disease. Thus, to understand both naturally occurring Wolbachia invasions and evaluate potential applied strategies, it is important to understand Wolbachia interactions with its host, including impacts on fitness and the CI level. In this study, we examined for an effect of Wolbachia on survivorship, developmental time, sex ratio, longevity, fecundity, and egg hatch of Aedes polynesiensis Marks, which is the primary vector of Wuchereria bancrofti in the South Pacific. In this study, we have compared strains of A. polynesiensis that are naturally and artificially infected with Wolbachia and additional strains that are aposymbiotic (Wolbachia removed to generate an uninfected strain). Artificially infected strains were observed to have increased larval mortality and decreased adult longevity when compared with aposymbiotic strains. Naturally infected strains were observed to have decreased larval mortality, pupal mortality, increased adult longevity, and a larger adult size when compared with aposymbiotic strains. Artificially infected males that were 4 wk old were able to induce high rates of CI, similar to young males. We discuss the results in relation to the natural spread of Wolbachia and Wolbachia-based applied strategies to modify A. polynesiensis populations.

  4. Influential sources affecting Bangkok adolescent body image perceptions.

    PubMed

    Thianthai, Chulanee

    2006-01-01

    The study of body image-related problems in non-Western countries is still very limited. Thus, this study aims to identify the main influential sources and show how they affect the body image perceptions of Bangkok adolescents. The researcher recruited 400 Thai male and female adolescents in Bangkok, attending high school to freshmen level, ranging from 16-19 years, to participate in this study. Survey questionnaires were distributed to every student and follow-up interviews conducted with 40 students. The findings showed that there are eight main influential sources respectively ranked from the most influential to the least influential: magazines, television, peer group, familial, fashion trend, the opposite gender, self-realization and health knowledge. Similar to those studies conducted in Western countries, more than half of the total percentage was the influence of mass media and peer groups. Bangkok adolescents also internalized Western ideal beauty through these mass media channels. Alike studies conducted in the West, there was similarities in the process of how these influential sources affect Bangkok adolescent body image perception, with the exception of familial source. In conclusion, taking the approach of identifying the main influential sources and understanding how they affect adolescent body image perceptions can help prevent adolescents from having unhealthy views and taking risky measures toward their bodies. More studies conducted in non-Western countries are needed in order to build a cultural sensitive program, catered to the body image problems occurring in adolescents within that particular society.

  5. Wolbachia wsp gene clones detect the distribution of Wolbachia variants and wsp hypervariable regions among individuals of a multistrain infected population of Diabrotica barberi (Coleoptera: Chrysomelidae)

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The northern corn rootworm (Diabrotica barberi) in eastern and central North America exhibits at least three distinct populations with respect to Wolbachia infection: uninfected; singly-infected; multi-infected. The infected states are associated with different mtDNA haplotypes and reduced mtDNA var...

  6. The symbiotic role of Wolbachia in Onchocercidae and its impact on filariasis.

    PubMed

    Bouchery, T; Lefoulon, E; Karadjian, G; Nieguitsila, A; Martin, C

    2013-02-01

    Symbiotic associations between eukaryotes and microorganisms are frequently observed in nature, and range along the continuum between parasitism and mutualism. The genus Wolbachia contains well-known intracellular bacteria of arthropods that induce several reproductive phenotypes that benefit the transmission of the bacteria. Interestingly, Wolbachia bacteria have been found in the Onchocercidae, a family of filarial nematodes, including species that cause human filarial diseases, e.g. lymphatic filariasis and onchocerciasis. The endosymbiont is thought to be mutualistic in the Onchocercidae, and to provide essential metabolites to the filariae. Currently, Wolbachia bacteria are targets of antibiotic therapy with tetracyclines, which have profound effects on the development, viability and fertility of filarial parasites. This overview article presents the Onchocercidae and Wolbachia, and then discusses the origin and the nature of the symbiosis. It highlights the contribution of Wolbachia to the survival of the filariae and to the development of pathology. Finally, the infection control implications for filariases are debated. Potential directions for future research are also discussed.

  7. Wolbachia Infections in Aedes aegypti Differ Markedly in Their Response to Cyclical Heat Stress

    PubMed Central

    Wiwatanaratanabutr, Itsanun; White, Vanessa L.; Hoffmann, Ary A.

    2017-01-01

    Aedes aegypti mosquitoes infected with Wolbachia bacteria are currently being released for arbovirus suppression around the world. Their potential to invade populations and persist will depend on interactions with environmental conditions, particularly as larvae are often exposed to fluctuating and extreme temperatures in the field. We reared Ae. aegypti larvae infected with different types of Wolbachia (wMel, wAlbB and wMelPop-CLA) under diurnal cyclical temperatures. Rearing wMel and wMelPop-CLA-infected larvae at 26–37°C reduced the expression of cytoplasmic incompatibility, a reproductive manipulation induced by Wolbachia. We also observed a sharp reduction in the density of Wolbachia in adults. Furthermore, the wMel and wMelPop-CLA infections were not transmitted to the next generation when mosquitoes were exposed to 26–37°C across all life stages. In contrast, the wAlbB infection was maintained at a high density, exhibited complete cytoplasmic incompatibility, and was transmitted from mother to offspring with a high fidelity under this temperature cycle. These findings have implications for the potential success of Wolbachia interventions across different environments and highlight the importance of temperature control in rearing. PMID:28056065

  8. Birth of a W sex chromosome by horizontal transfer of Wolbachia bacterial symbiont genome

    PubMed Central

    Leclercq, Sébastien; Thézé, Julien; Chebbi, Mohamed Amine; Giraud, Isabelle; Moumen, Bouziane; Ernenwein, Lise; Grève, Pierre; Cordaux, Richard

    2016-01-01

    Sex determination is a fundamental developmental pathway governing male and female differentiation, with profound implications for morphology, reproductive strategies, and behavior. In animals, sex differences between males and females are generally determined by genetic factors carried by sex chromosomes. Sex chromosomes are remarkably variable in origin and can differ even between closely related species, indicating that transitions occur frequently and independently in different groups of organisms. The evolutionary causes underlying sex chromosome turnover are poorly understood, however. Here we provide evidence indicating that Wolbachia bacterial endosymbionts triggered the evolution of new sex chromosomes in the common pillbug Armadillidium vulgare. We identified a 3-Mb insert of a feminizing Wolbachia genome that was recently transferred into the pillbug nuclear genome. The Wolbachia insert shows perfect linkage to the female sex, occurs in a male genetic background (i.e., lacking the ancestral W female sex chromosome), and is hemizygous. Our results support the conclusion that the Wolbachia insert is now acting as a female sex-determining region in pillbugs, and that the chromosome carrying the insert is a new W sex chromosome. Thus, bacteria-to-animal horizontal genome transfer represents a remarkable mechanism underpinning the birth of sex chromosomes. We conclude that sex ratio distorters, such as Wolbachia endosymbionts, can be powerful agents of evolutionary transitions in sex determination systems in animals. PMID:27930295

  9. Wolbachia Infections in Aedes aegypti Differ Markedly in Their Response to Cyclical Heat Stress.

    PubMed

    Ross, Perran A; Wiwatanaratanabutr, Itsanun; Axford, Jason K; White, Vanessa L; Endersby-Harshman, Nancy M; Hoffmann, Ary A

    2017-01-01

    Aedes aegypti mosquitoes infected with Wolbachia bacteria are currently being released for arbovirus suppression around the world. Their potential to invade populations and persist will depend on interactions with environmental conditions, particularly as larvae are often exposed to fluctuating and extreme temperatures in the field. We reared Ae. aegypti larvae infected with different types of Wolbachia (wMel, wAlbB and wMelPop-CLA) under diurnal cyclical temperatures. Rearing wMel and wMelPop-CLA-infected larvae at 26-37°C reduced the expression of cytoplasmic incompatibility, a reproductive manipulation induced by Wolbachia. We also observed a sharp reduction in the density of Wolbachia in adults. Furthermore, the wMel and wMelPop-CLA infections were not transmitted to the next generation when mosquitoes were exposed to 26-37°C across all life stages. In contrast, the wAlbB infection was maintained at a high density, exhibited complete cytoplasmic incompatibility, and was transmitted from mother to offspring with a high fidelity under this temperature cycle. These findings have implications for the potential success of Wolbachia interventions across different environments and highlight the importance of temperature control in rearing.

  10. Wolbachia restricts insect-specific flavivirus infection in Aedes aegypti cells

    PubMed Central

    Sreenu, Vatipally B.; Mottram, Timothy; McFarlane, Melanie

    2016-01-01

    Mosquito-borne viruses are known to cause disease in humans and livestock and are often difficult to control due to the lack of specific antivirals and vaccines. The Wolbachia endosymbiont has been widely studied for its ability to restrict positive-strand RNA virus infection in mosquitoes, although little is known about the precise antiviral mechanism. In recent years, a variety of insect-specific viruses have been discovered in mosquitoes and an interaction with mosquito-borne viruses has been reported for some of them; however, nothing is known about the effect of Wolbachia on insect-specific virus infection in mosquitoes. Here, we show that transinfection of the Drosophila-derived wMelPop Wolbachia strain into Aedes aegypti-derived cells resulted in inhibition and even clearance of the persistent cell-fusing agent flavivirus infection in these cells. This broadens the antiviral activity of Wolbachia from acute infections to persistent infections and from arboviruses to mosquito-specific viruses. In contrast, no effect on the Phasi Charoen-like bunyavirus persistent infection in these cells was observed, suggesting a difference in Wolbachia inhibition between positive- and negative-strand RNA viruses. PMID:27692043

  11. Successful establishment of Wolbachia in Aedes populations to suppress dengue transmission.

    PubMed

    Hoffmann, A A; Montgomery, B L; Popovici, J; Iturbe-Ormaetxe, I; Johnson, P H; Muzzi, F; Greenfield, M; Durkan, M; Leong, Y S; Dong, Y; Cook, H; Axford, J; Callahan, A G; Kenny, N; Omodei, C; McGraw, E A; Ryan, P A; Ritchie, S A; Turelli, M; O'Neill, S L

    2011-08-24

    Genetic manipulations of insect populations for pest control have been advocated for some time, but there are few cases where manipulated individuals have been released in the field and no cases where they have successfully invaded target populations. Population transformation using the intracellular bacterium Wolbachia is particularly attractive because this maternally-inherited agent provides a powerful mechanism to invade natural populations through cytoplasmic incompatibility. When Wolbachia are introduced into mosquitoes, they interfere with pathogen transmission and influence key life history traits such as lifespan. Here we describe how the wMel Wolbachia infection, introduced into the dengue vector Aedes aegypti from Drosophila melanogaster, successfully invaded two natural A. aegypti populations in Australia, reaching near-fixation in a few months following releases of wMel-infected A. aegypti adults. Models with plausible parameter values indicate that Wolbachia-infected mosquitoes suffered relatively small fitness costs, leading to an unstable equilibrium frequency <30% that must be exceeded for invasion. These findings demonstrate that Wolbachia-based strategies can be deployed as a practical approach to dengue suppression with potential for area-wide implementation.

  12. From Parasite to Mutualist: Rapid Evolution of Wolbachia in Natural Populations of Drosophila

    PubMed Central

    Weeks, Andrew R; Turelli, Michael; Harcombe, William R; Reynolds, K. Tracy; Hoffmann, Ary A

    2007-01-01

    Wolbachia are maternally inherited bacteria that commonly spread through host populations by causing cytoplasmic incompatibility, often expressed as reduced egg hatch when uninfected females mate with infected males. Infected females are frequently less fecund as a consequence of Wolbachia infection. However, theory predicts that because of maternal transmission, these “parasites” will tend to evolve towards a more mutualistic association with their hosts. Drosophila simulans in California provided the classic case of a Wolbachia infection spreading in nature. Cytoplasmic incompatibility allowed the infection to spread through individual populations within a few years and from southern to northern California (more than 700 km) within a decade, despite reducing the fecundity of infected females by 15%–20% under laboratory conditions. Here we show that the Wolbachia in California D. simulans have changed over the last 20 y so that infected females now exhibit an average 10% fecundity advantage over uninfected females in the laboratory. Our data suggest smaller but qualitatively similar changes in relative fecundity in nature and demonstrate that fecundity-increasing Wolbachia variants are currently polymorphic in natural populations. PMID:17439303

  13. Comparative analysis of Wolbachia surface protein in D. melanoagster, A. tabida and B. malayi

    PubMed Central

    Uday, Jayaramaiah; Puttaraju, Hosagavi Puttegowda

    2012-01-01

    Wolbachia surface protein (WSP) is an eight beta-barrel transmembrane structure which participates in host immune response, cell proliferation, pathogenicity and controlled cell death program. The protein has four extracellular loops containing hyper variable regions separated by conserved regions. The WSP structure is homologous to Neisseria surface protein (Nsp A) which has about 34% similarity including antigenic variation and hydrophilicity. Recombination has a large impact on diversity of this protein including positive selection which is major constraint on protein evolution. The molecular mechanism through which Wolbachia induces various reproductive anomalies is unclear; a key feature observed for such anomalies might be because of Wolbachia undergoing extensive recombination. In Wolbachia, increased recombination is observed in ankyrin proteins, surface proteins and in some hypothetical proteins. Genetic divergence is extensive in the WSP gene, WSP is known to be a chimeric protein involved in host-symbiont interactions. Here we predicted the structural and functional variations in WSP sequences of Wolbachia present in D. melanogaster, A. tabida and in B. malayi. PMID:23055615

  14. Wolbachia in European Populations of the Invasive Pest Drosophila suzukii: Regional Variation in Infection Frequencies

    PubMed Central

    Gibert, Patricia; Martinez, Julien; Fraimout, Antoine; Jiggins, Francis; Andrieux, Thibault; Siozios, Stefanos; Anfora, Gianfranco; Miller, Wolfgang; Rota-Stabelli, Omar; Mouton, Laurence

    2016-01-01

    The invasive pest Drosophila suzukii is characterized by a specific fresh-fruit targeting behavior and has quickly become a menace for the fruit economy of newly infested North American and European regions. D. suzukii carries a strain of the endosymbiotic bacterium Wolbachia, named wSuz, which has a low infection frequency and no reproductive manipulation capabilities in American populations of D. suzukii. To further understand the nature of wSuz biology and assess its utility as a tool for controlling this pest’s populations, we investigated the prevalence of Wolbachia in 23 European D. suzukii populations, and compared our results with those available in American populations. Our data showed a highly variable infection frequency with a mean prevalence of 46%, which is significantly higher than the 17% found in American populations. Based on Multilocus Sequence Typing analysis, a single wSuz strain was diagnosed in all European populations of D. suzukii. In agreement with American data, we found no evidence of cytoplasmic incompatibility induced by wSuz. These findings raise two questions: a) why Wolbachia is maintained in field populations of D. suzukii and b) what are the selective forces responsible for the variation in prevalence within populations, particularly between European and American continents? Our results provide new insights into the D. suzukii-Wolbachia association and highlight regional variations that await further investigation and that should be taken into account for using Wolbachia-based pest management programs. PMID:26809119

  15. Reproductive compensation favours male-killing Wolbachia in a live-bearing host

    PubMed Central

    Koop, Julie L.; Zeh, David W.; Bonilla, Melvin M.; Zeh, Jeanne A.

    2009-01-01

    Wolbachia are maternally inherited, cellular endosymbionts that can enhance their fitness by biasing host sex ratio in favour of females. Male killing (MK) is an extreme form of sex-ratio manipulation that is selectively advantageous if the self-sacrifice of Wolbachia in males increases transmission through females. In live-bearing hosts, females typically produce more embryos than can be carried to term, and reproductive compensation through maternal resource reallocation from dead males to female embryos could increase the number of daughters born to infected females. Here, we report a new strain of MK Wolbachia (wCsc2) in the pseudoscorpion, Cordylochernes scorpioides, and present the first empirical evidence that reproductive compensation favours the killing of males in a viviparous host. Females infected with the wCsc2 strain produced 26 per cent more and significantly larger daughters than tetracycline-cured females. In contrast to the previously described wCsc1 MK Wolbachia strain in C. scorpioides, wCsc2 infection was not accompanied by an increase in the rate of spontaneous brood abortion. Characterization of the wCsc1 and wCsc2 strains by multi-locus sequence typing and by Wolbachia surface protein (wsp) gene sequencing indicates that the marked divergence between these two MK strains in their impact on host reproductive success, and hence in their potential to spread, has occurred in association with homologous recombination in the wsp gene. PMID:19710065

  16. Wolbachia infections in mosquitoes and their predators inhabiting rice field communities in Thailand and China.

    PubMed

    Wiwatanaratanabutr, Itsanun; Zhang, Chongxing

    2016-07-01

    Wolbachia are inherited, endocytoplasmic bacteria that infect a wide range of arthropods. Here is the first systematic report on the study of Wolbachia infection in mosquitoes and their predators from both Thailand and China. In Thailand, 632 mosquito specimens (20 spp.) and 424 insect predators (23 spp.) were collected from the rice agroecosystem, mostly from the Central region, followed by the Northeast, the North and the South and were inhabiting rice fields, wetlands and ditches. In China, 928 mosquitoes (15 spp.) and 149 insect predators (16 spp.) were collected from rice fields along the Weishan Lake in Shandong province. Specimens were classified in the orders Diptera, Coleoptera, Odonata and Hemiptera. Using wsp, ftsZ, 16S rRNA and groE gene amplifications, Wolbachia were detected in 12 mosquito spp. and 6 predator spp. from Thailand and 11 mosquito spp. and 5 predator spp. from China. The relative Wolbachia densities of these species were determined using quantitative real-time PCR. The mosquito, Aedes albopictus, and the predator, Agriocnemis femina, had the highest bacterial densities. These results imply that Wolbachia of supergroup B are distributed throughout these insects, probably via horizontal transmission in rice agroecosystems.

  17. Detection of Low-Level Cardinium and Wolbachia Infections in Culicoides.

    PubMed

    Mee, Peter T; Weeks, Andrew R; Walker, Peter J; Hoffmann, Ary A; Duchemin, Jean-Bernard

    2015-09-01

    Bacterial endosymbionts have been identified as potentially useful biological control agents for a range of invertebrate vectors of disease. Previous studies of Culicoides (Diptera: Ceratopogonidae) species using conventional PCR assays have provided evidence of Wolbachia (1/33) and Cardinium (8/33) infections. Here, we screened 20 species of Culicoides for Wolbachia and Cardinium, utilizing a combination of conventional PCR and more sensitive quantitative PCR (qPCR) assays. Low levels of Cardinium DNA were detected in females of all but one of the Culicoides species screened, and low levels of Wolbachia were detected in females of 9 of the 20 Culicoides species. Sequence analysis based on partial 16S rRNA gene and gyrB sequences identified "Candidatus Cardinium hertigii" from group C, which has previously been identified in Culicoides from Japan, Israel, and the United Kingdom. Wolbachia strains detected in this study showed 98 to 99% sequence identity to Wolbachia previously detected from Culicoides based on the 16S rRNA gene, whereas a strain with a novel wsp sequence was identified in Culicoides narrabeenensis. Cardinium isolates grouped to geographical regions independent of the host Culicoides species, suggesting possible geographical barriers to Cardinium movement. Screening also identified Asaia bacteria in Culicoides. These findings point to a diversity of low-level endosymbiont infections in Culicoides, providing candidates for further characterization and highlighting the widespread occurrence of these endosymbionts in this insect group.

  18. Wolbachia in European Populations of the Invasive Pest Drosophila suzukii: Regional Variation in Infection Frequencies.

    PubMed

    Cattel, Julien; Kaur, Rupinder; Gibert, Patricia; Martinez, Julien; Fraimout, Antoine; Jiggins, Francis; Andrieux, Thibault; Siozios, Stefanos; Anfora, Gianfranco; Miller, Wolfgang; Rota-Stabelli, Omar; Mouton, Laurence

    2016-01-01

    The invasive pest Drosophila suzukii is characterized by a specific fresh-fruit targeting behavior and has quickly become a menace for the fruit economy of newly infested North American and European regions. D. suzukii carries a strain of the endosymbiotic bacterium Wolbachia, named wSuz, which has a low infection frequency and no reproductive manipulation capabilities in American populations of D. suzukii. To further understand the nature of wSuz biology and assess its utility as a tool for controlling this pest's populations, we investigated the prevalence of Wolbachia in 23 European D. suzukii populations, and compared our results with those available in American populations. Our data showed a highly variable infection frequency with a mean prevalence of 46%, which is significantly higher than the 17% found in American populations. Based on Multilocus Sequence Typing analysis, a single wSuz strain was diagnosed in all European populations of D. suzukii. In agreement with American data, we found no evidence of cytoplasmic incompatibility induced by wSuz. These findings raise two questions: a) why Wolbachia is maintained in field populations of D. suzukii and b) what are the selective forces responsible for the variation in prevalence within populations, particularly between European and American continents? Our results provide new insights into the D. suzukii-Wolbachia association and highlight regional variations that await further investigation and that should be taken into account for using Wolbachia-based pest management programs.

  19. Absence of Wolbachia endobacteria in Sri Lankan isolates of the nematode parasite of animals Setaria digitata.

    PubMed

    Voronin, Denis; Abeykoon, A M L L; Gunawardene, Y I Silva; Dassanayake, Ranil S

    2015-01-30

    Setaria digitata is an animal filarial parasite with natural hosts of cattle and buffaloes that causes mild disease conditions. Infection of non-permissive hosts such as goats, sheep and horses, by this nematode can cause cerebrospinal nematodiasis that leads to lumbar paralysis and the eventual death of the animals and inflicts considerable economic losses on livestock farmers. Wolbachia are obligate mutualistic endosymbionts for some filarial nematodes and are currently being targeted for the control of diseases caused by these parasites. However, little is known about the occurrence of this endosymbiont in the Setariidae family. In this work, worms collected from infected cattle in Sri Lanka were morphologically identified as S. digitata and tested for the presence of Wolbachia by PCR screening using the WSP- and Wolbachia-specific 16S rRNA and multilocus sequence typing primers that were designed to amplify the gatB, coxA, hcpA, ftsZ and fbpA sequences of Wolbachia. The presence of endobacteria in S. digitata was also examined by whole-mount immunofluorescence staining of the parasites and transmission electron microscopic studies. These analyses did not produce evidence of presence of Wolbachia or any other endosymbiotic bacteria in S. digitata, whereas such evidence was found in Brugia malayi, which was used as a positive control in this study.

  20. Detection of Low-Level Cardinium and Wolbachia Infections in Culicoides

    PubMed Central

    Mee, Peter T.; Weeks, Andrew R.; Walker, Peter J.; Hoffmann, Ary A.

    2015-01-01

    Bacterial endosymbionts have been identified as potentially useful biological control agents for a range of invertebrate vectors of disease. Previous studies of Culicoides (Diptera: Ceratopogonidae) species using conventional PCR assays have provided evidence of Wolbachia (1/33) and Cardinium (8/33) infections. Here, we screened 20 species of Culicoides for Wolbachia and Cardinium, utilizing a combination of conventional PCR and more sensitive quantitative PCR (qPCR) assays. Low levels of Cardinium DNA were detected in females of all but one of the Culicoides species screened, and low levels of Wolbachia were detected in females of 9 of the 20 Culicoides species. Sequence analysis based on partial 16S rRNA gene and gyrB sequences identified “Candidatus Cardinium hertigii” from group C, which has previously been identified in Culicoides from Japan, Israel, and the United Kingdom. Wolbachia strains detected in this study showed 98 to 99% sequence identity to Wolbachia previously detected from Culicoides based on the 16S rRNA gene, whereas a strain with a novel wsp sequence was identified in Culicoides narrabeenensis. Cardinium isolates grouped to geographical regions independent of the host Culicoides species, suggesting possible geographical barriers to Cardinium movement. Screening also identified Asaia bacteria in Culicoides. These findings point to a diversity of low-level endosymbiont infections in Culicoides, providing candidates for further characterization and highlighting the widespread occurrence of these endosymbionts in this insect group. PMID:26150447

  1. The microbiome composition of Aedes aegypti is not critical for Wolbachia-mediated inhibition of dengue virus

    PubMed Central

    Audsley, Michelle D.; Ye, Yixin H.

    2017-01-01

    Background Dengue virus (DENV) is primarily vectored by the mosquito Aedes aegypti, and is estimated to cause 390 million human infections annually. A novel method for DENV control involves stable transinfection of Ae. aegypti with the common insect endosymbiont Wolbachia, which mediates an antiviral effect. However, the mechanism by which Wolbachia reduces the susceptibility of Ae. aegypti to DENV is not fully understood. In this study we assessed the potential of resident microbiota, which can play important roles in insect physiology and immune responses, to affect Wolbachia-mediated DENV blocking. Methodology/Findings The microbiome of Ae. aegypti stably infected with Wolbachia strain wMel was compared to that of Ae. aegypti without Wolbachia, using 16s rDNA profiling. Our results indicate that although Wolbachia affected the relative abundance of several genera, the microbiome of both the Wolbachia-infected and uninfected mosquitoes was dominated by Elizabethkingia and unclassified Enterobacteriaceae. To assess the potential of the resident microbiota to affect the Wolbachia-mediated antiviral effect, we used antibiotic treatment before infection with DENV by blood-meal. In spite of a significant shift in the microbiome composition in response to the antibiotics, we detected no effect of antibiotic treatment on DENV infection rates, or on the DENV load of infected mosquitoes. Conclusions/Significance Our findings indicate that stable infection with Wolbachia strain wMel produces few effects on the microbiome of laboratory-reared Ae. aegypti. Moreover, our findings suggest that the microbiome can be significantly altered without affecting the fundamental DENV blocking phenotype in these mosquitoes. Since Ae. aegypti are likely to encounter diverse microbiota in the field, this is a particularly important result in the context of using Wolbachia as a method for DENV control. PMID:28267749

  2. A new case of Wolbachia dependence in the genus Asobara: evidence for parthenogenesis induction in Asobara japonica.

    PubMed

    Kremer, N; Charif, D; Henri, H; Bataille, M; Prévost, G; Kraaijeveld, K; Vavre, F

    2009-09-01

    Wolbachia is a maternally inherited bacterium that is widely distributed among arthropods, in which it manipulates the reproduction of its hosts. Although generally facultative for its hosts, Wolbachia has recently become obligatory in Asobara tabida (Hymenoptera: Braconidae) in which it is required for the completion of oogenesis. Here, we describe a new Wolbachia strain (wAjap) that is associated with the genus Asobara and infects Asobara japonica. wAjap was detected in all female-biased populations of A. japonica found in the main islands of Japan, but not in the arrhenotokous populations from the southern islands. Using phylogenetic analyses based on multi-locus sequence typing (MLST), we show that this strain is closely related to wAtab3 (the strain required for oogenesis in A. tabida), even though they differ on Wolbachia surface protein (WSP) and WO phage sequences. Using antibiotic treatments, we show that cured thelytokous females are not dependent on Wolbachia for oogenesis. However, they produced only sons, showing that wAjap induces thelytokous parthenogenesis. Analyses of mating behavior and offspring production of individuals from Wolbachia-infected populations showed that while males were still sexually functional, females no longer attract males, making Wolbachia an obligate partner for daughter production in thelytokous populations. The fact that Wolbachia has become independently obligatory in two species of the same genus tends to show that dependence evolution can be common and swift, although no clear benefit for the parasitoid can be attributed to this dependence. Although dependence should lead to co-divergence between Wolbachia and its hosts, the very few cases of co-speciation observed in host-Wolbachia associations question the stability of these obligatory associations.

  3. Wolbachia endosymbiont of Brugia malayi elicits a T helper type 17-mediated pro-inflammatory immune response through Wolbachia surface protein.

    PubMed

    Pathak, Manisha; Verma, Meenakshi; Srivastava, Mrigank; Misra-Bhattacharya, Shailja

    2015-02-01

    Wolbachia is an endosymbiotic bacterium of the filarial nematode Brugia malayi. The symbiotic relationship between Wolbachia and its filarial host is dependent on interactions between the proteins of both organisms. However, little is known about Wolbachia proteins that are involved in the inflammatory pathology of the host during lymphatic filariasis. In the present study, we cloned, expressed and purified Wolbachia surface protein (r-wsp) from Wolbachia and administered it to mice, either alone or in combination with infective larvae of B. malayi (Bm-L3) and monitored the developing immune response in infected animals. Our results show that spleens and mesenteric lymph nodes of mice immunized with either r-wsp or infected with Bm-L3 show increased percentages of CD4(+) T helper type 17 (Th17) cells and Th1 cytokines like interferon-γ and interleukin-2 (IL-2) along with decreased percentages of regulatory T cells, Th2 cytokines like IL-4 and IL-10 and transforming growth factor β (TGF-β) levels in culture supernatants of splenocytes. These observations were stronger in mice immunized with r-wsp alone. Interestingly, when mice were first immunized with r-wsp and subsequently infected with Bm-L3, percentages of CD4(+) Th17 cells and Th1 cytokines increased even further while that of regulatory T cells, Th2 cytokines and TGF-β levels decreased. These results for the first time show that r-wsp acts synergistically with Bm-L3 in promoting a pro-inflammatory response by increasing Th17 cells and at the same time diminishes host immunological tolerance by decreasing regulatory T cells and TGF-β secretion.

  4. Convergent incidences of Wolbachia infection in fig wasp communities from two continents

    PubMed Central

    Haine, Eleanor R.; Cook, James M.

    2005-01-01

    Wide surveys suggest that Wolbachia bacteria infect ca. 20% of all insect species, but particular taxonomic or ecological groups may display significantly higher or lower incidences. We studied 61 fig wasp species in Australia and found the highest known incidence (67%) of infection in a targeted study of this nature. A comparable study in Panama reported a similar figure (59%), confirming the exceptionally high incidence of Wolbachia in fig wasps. Importantly, these are two independent estimates of Wolbachia incidence in fig wasp communities, because no host species, or even genera, are shared between localities. The high level of infection may reflect enhanced opportunities for horizontal transmission inside fig fruits. Although incidence was similar in Panama and Australia, the actual strains involved were different and region-specific. Local strains were shared by several host species, although there was often no obvious (direct) ecological link between two hosts with the same infection. PMID:15734697

  5. Tripartite associations among bacteriophage WO, Wolbachia, and host affected by temperature and age in Tetranychus urticae.

    PubMed

    Lu, Ming-Hong; Zhang, Kai-Jun; Hong, Xiao-Yue

    2012-11-01

    A phage density model of cytoplasmic incompatibility (CI), which means lytic phages reduce bacterial density associated with CI, significantly enhances our understanding of the tripartite associations among bacteriophage WO, Wolbachia and host. However, WO may alternate between lytic and lysogenic life cycles or change phage production under certain conditions including temperature, host age and host species background. Here, extreme temperatures can induce an alteration in the life cycle of WO and change the tripartite associations among WO, Wolbachia and CI. Based on the accumulation of the WO load, WO can transform into the lytic life cycle with increasing age. These findings confirmed that the environment plays an important role in the associations among WO, Wolbachia and host.

  6. Effects of mimosine on Wolbachia in mosquito cells: cell cycle suppression reduces bacterial abundance.

    PubMed

    Fallon, Ann M

    2015-10-01

    The plant allelochemical L-mimosine (β-[N-(3-hydroxy-4-pyridone)]-α-aminopropionic acid; leucenol) resembles the nonessential amino acid, tyrosine. Because the obligate intracellular alphaproteobacterium, Wolbachia pipientis, metabolizes amino acids derived from host cells, the effects of mimosine on infected and uninfected mosquito cells were investigated. The EC50 for mimosine was 6-7 μM with Aedes albopictus C7-10 and C/wStr cell lines, and was not influenced by infection status. Mosquito cells responded to concentrations of mimosine substantially lower than those used to synchronize the mammalian cell cycle; at concentrations of 30-35 μM, mimosine reversibly arrested the mosquito cell cycle at the G1/S boundary and inhibited growth of Wolbachia strain wStr. Although lower concentrations of mimosine slightly increased wStr abundance, concentrations that suppressed the cell cycle reduced Wolbachia levels.

  7. The potential of virulent Wolbachia to modulate disease transmission by insects.

    PubMed

    Brownstein, J S; Hett, E; O'Neill, S L

    2003-09-01

    A virulent strain of Wolbachia has recently been identified in Drosophila that drastically reduces adult lifespan. It has been proposed that this phenotype might be introduced into insect disease vector populations to reduce pathogen transmission. Here we model the requirements for spread of such an agent and the associated reduction in disease transmission. First, a simulation of mosquito population age structure was used to describe the age distribution of mosquitoes transmitting dengue virus. Second, given varying levels of cytoplasmic incompatibility and fecundity effect, the maximum possible longevity reduction that would allow Wolbachia to invade was obtained. Finally, the two models were combined to estimate the reduction in disease transmission according to different introduction frequencies. With strong CI and limited effect of fecundity, an introduction of Wolbachia with an initial frequency of 0.4 could result in a 60-80% reduction of transmitting mosquitoes. Greater reductions are possible at higher initial release rates.

  8. Defining Brugia malayi and Wolbachia symbiosis by stage-specific dual RNA-seq

    PubMed Central

    Voronin, Denis; Ding, Tao; Twaddle, Alan; Unnasch, Thomas R.; Lustigman, Sara; Ghedin, Elodie

    2017-01-01

    Background Filarial nematodes currently infect up to 54 million people worldwide, with millions more at risk for infection, representing the leading cause of disability in the developing world. Brugia malayi is one of the causative agents of lymphatic filariasis and remains the only human filarial parasite that can be maintained in small laboratory animals. Many filarial nematode species, including B. malayi, carry an obligate endosymbiont, the alpha-proteobacteria Wolbachia, which can be eliminated through antibiotic treatment. Elimination of the endosymbiont interferes with development, reproduction, and survival of the worms within the mamalian host, a clear indicator that the Wolbachia are crucial for survival of the parasite. Little is understood about the mechanism underlying this symbiosis. Methodology/ Principle findings To better understand the molecular interplay between these two organisms we profiled the transcriptomes of B. malayi and Wolbachia by dual RNA-seq across the life cycle of the parasite. This helped identify functional pathways involved in this essential symbiotic relationship provided by the co-expression of nematode and bacterial genes. We have identified significant stage-specific and gender-specific differential expression in Wolbachia during the nematode’s development. For example, during female worm development we find that Wolbachia upregulate genes involved in ATP production and purine biosynthesis, as well as genes involved in the oxidative stress response. Conclusions/ Significance This global transcriptional analysis has highlighted specific pathways to which both Wolbachia and B. malayi contribute concurrently over the life cycle of the parasite, paving the way for the development of novel intervention strategies. PMID:28358880

  9. Feminizing Wolbachia: a transcriptomics approach with insights on the immune response genes in Armadillidium vulgare

    PubMed Central

    2012-01-01

    Background Wolbachia are vertically transmitted bacteria known to be the most widespread endosymbiont in arthropods. They induce various alterations of the reproduction of their host, including feminization of genetic males in isopod crustaceans. In the pill bug Armadillidium vulgare, the presence of Wolbachia is also associated with detrimental effects on host fertility and lifespan. Deleterious effects have been demonstrated on hemocyte density, phenoloxidase activity, and natural hemolymph septicemia, suggesting that infected individuals could have defective immune capacities. Since nothing is known about the molecular mechanisms involved in Wolbachia-A. vulgare interactions and its secondary immunocompetence modulation, we developed a transcriptomics strategy and compared A. vulgare gene expression between Wolbachia-infected animals (i.e., “symbiotic” animals) and uninfected ones (i.e., “asymbiotic” animals) as well as between animals challenged or not challenged by a pathogenic bacteria. Results Since very little genetic data is available on A. vulgare, we produced several EST libraries and generated a total of 28 606 ESTs. Analyses of these ESTs revealed that immune processes were over-represented in most experimental conditions (responses to a symbiont and to a pathogen). Considering canonical crustacean immune pathways, these genes encode antimicrobial peptides or are involved in pathogen recognition, detoxification, and autophagy. By RT-qPCR, we demonstrated a general trend towards gene under-expression in symbiotic whole animals and ovaries whereas the same gene set tends to be over-expressed in symbiotic immune tissues. Conclusion This study allowed us to generate the first reference transcriptome ever obtained in the Isopoda group and to identify genes involved in the major known crustacean immune pathways encompassing cellular and humoral responses. Expression of immune-related genes revealed a modulation of host immunity when females are

  10. What can symbiont titres tell us about co-evolution of Wolbachia and their host?

    PubMed

    Correa, C Carolina; Ballard, J William O

    2014-05-01

    There is a long-standing prediction that associations with vertically transmitted symbionts evolve towards maximisation of host reproductive success, eventually leading to mutualist symbiosis and coadaptation. Under this scenario, the regulation of symbiont titres in host tissues would be expected to be greater when partners have coevolved for a long time than when they have recently met. Wolbachia pipientis, a common vertically transmitted symbiont of invertebrates, often has the capacity to spread through the host population without being beneficial to the hosts, by means of reducing the hatch rate in crosses between uninfected females and infected males. This manipulation, namely cytoplasmic incompatibility (CI), may exert strong selection on the accuracy of infection transmission from mother to offspring, and therefore, on regulation of symbiont titres in the ova. Here, we examined the symbiont density dynamics in gonads of Drosophila simulans infected with the wMa strain of Wolbachia, known to cause mild CI and likely to be the oldest Wolbachia infection known to this fly species. Further, we compared these results with those obtained for the more recent association between D. simulans and the potent CI-inducer wHa (Correa and Ballard, 2012). We aimed to determine if the regulation of Wolbachia density in fly gonads is greater in the older association, as would be predicted solely by gradual coadaptation, or if the selection exerted by CI on reproductive fitness could also play a role, therefore showing tighter regulation on flies with the stronger CI-inducing strain. We observed that Wolbachia density in gonads of wMa infected flies changed with laboratory adaptation and were disturbed by environmental challenges, which contrasted with the stability of ovarian wHa density to the same treatments. Our observations are in line with the prediction that selection on reproductive fitness influences the evolution symbiont density regulation in Drosophila, and may

  11. Stable coexistence of incompatible Wolbachia along a narrow contact zone in mosquito field populations.

    PubMed

    Atyame, Célestine M; Labbé, Pierrick; Rousset, François; Beji, Marwa; Makoundou, Patrick; Duron, Olivier; Dumas, Emilie; Pasteur, Nicole; Bouattour, Ali; Fort, Philippe; Weill, Mylène

    2015-01-01

    In arthropods, the intracellular bacteria Wolbachia often induce cytoplasmic incompatibility (CI) between sperm and egg, which causes conditional embryonic death and promotes the spatial spread of Wolbachia infections into host populations. The ability of Wolbachia to spread in natural populations through CI has attracted attention for using these bacteria in vector-borne disease control. The dynamics of incompatible Wolbachia infections have been deeply investigated theoretically, whereas in natural populations, there are only few examples described, especially among incompatible infected hosts. Here, we have surveyed the distribution of two molecular Wolbachia strains (wPip11 and wPip31) infecting the mosquito Culex pipiens in Tunisia. We delineated a clear spatial structure of both infections, with a sharp contact zone separating their distribution areas. Crossing experiments with isofemale lines from different localities showed three crossing types: wPip11-infected males always sterilize wPip31-infected females; however, while most wPip31-infected males were compatible with wPip11-infected females, a few completely sterilize them. The wPip11 strain was thus expected to spread, but temporal dynamics over 7 years of monitoring shows the stability of the contact zone. We examined which factors may contribute to the observed stability, both theoretically and empirically. Population cage experiments, field samples and modelling did not support significant impacts of local adaptation or assortative mating on the stability of wPip infection structure. By contrast, low dispersal probability and metapopulation dynamics in the host Cx. pipiens probably play major roles. This study highlights the need of understanding CI dynamics in natural populations to design effective and sustainable Wolbachia-based control strategies.

  12. Medical guidelines for space passengers--II.

    PubMed

    Rayman, Russell B; Antuñano, Melchor J; Garber, Mitchell A; Hastings, John D; Illig, Petra A; Jordan, Jon L; Landry, Roger F; McMeekin, Robert R; Northrup, Susan E; Ruehle, Charles; Saenger, Arleen; Schneider, Victor S

    2002-11-01

    It now appears likely that commercial entities will carry paying passengers on suborbital spaceflights in this decade. The stresses of spaceflight, the effects of microgravity, and the limited capability for medical care onboard make it advisable to develop a system of medical clearance for such space tourists. The Aerospace Medical Association, therefore, organized a Space Passenger Task Force whose first report on medical guidelines was published in 2001. That report consisted of a list of conditions that would disqualify potential passengers for relatively long orbital flights. The Task Force reconvened in 2002 to focus on less stringent medical screening appropriate for short duration suborbital flights. It was assumed that such commercial flights would involve: 1) small spacecraft carrying 4-6 passengers; 2) a cabin maintained at sea-level "shirt-sleeve" condition; 3) maximum accelerations of 2.0-4.5 G; 4) about 30 min in microgravity. The Task Force addressed specific medical problems, including space motion sickness, pregnancy, and medical conditions involving the risk of sudden incapacitation. The Task Force concluded that a medical history should be taken from potential passengers with individualized follow-up that focuses on areas of concern.

  13. Identifying influential spreaders in complex networks based on gravity formula

    NASA Astrophysics Data System (ADS)

    Ma, Ling-ling; Ma, Chuang; Zhang, Hai-Feng; Wang, Bing-Hong

    2016-06-01

    How to identify the influential spreaders in social networks is crucial for accelerating/hindering information diffusion, increasing product exposure, controlling diseases and rumors, and so on. In this paper, by viewing the k-shell value of each node as its mass and the shortest path distance between two nodes as their distance, then inspired by the idea of the gravity formula, we propose a gravity centrality index to identify the influential spreaders in complex networks. The comparison between the gravity centrality index and some well-known centralities, such as degree centrality, betweenness centrality, closeness centrality, and k-shell centrality, and so forth, indicates that our method can effectively identify the influential spreaders in real networks as well as synthetic networks. We also use the classical Susceptible-Infected-Recovered (SIR) epidemic model to verify the good performance of our method.

  14. A Native Wolbachia Endosymbiont Does Not Limit Dengue Virus Infection in the Mosquito Aedes notoscriptus (Diptera: Culicidae).

    PubMed

    Skelton, Ellie; Rancès, Edwige; Frentiu, Francesca D; Kusmintarsih, Endang Srimurni; Iturbe-Ormaetxe, Iñaki; Caragata, Eric P; Woolfit, Megan; O'Neill, Scott L

    2016-03-01

    The endosymbiotic bacterium Wolbachia pipientis infects many species of insects and has been transinfected into the mosquito Aedes aegypti (L.), the primary vector of dengue virus (DENV). Recently, it has been shown that Wolbachia blocks the replication and transmission of RNA viruses, such as DENV, in a number of mosquito species including Ae. aegypti and Aedes albopictus (Skuse), which is naturally infected with Wolbachia and considered a secondary vector for DENV. The mosquito species Aedes notoscriptus (Skuse) is highly prevalent in Australia, including in areas where DENV outbreaks have been recorded. The mosquito has been implicated in the transmission of Ross River and Barmah Forest viruses, but not DENV. We investigated whether Wolbachia naturally infects this mosquito species and whether it has an impact on the ability of Ae. notoscriptus to transmit DENV. We show, for the first time, that Ae. notoscriptus is naturally infected with a strain of Wolbachia that belongs to supergroup B and is localized only in the ovaries. However, Wolbachia infection in Ae. notoscriptus did not induce resistance to DENV and had no effect on overall DENV infection rate or titer. The presence of a native Wolbachia in Ae. notoscriptus cannot explain why this mosquito is an ineffective vector of DENV.

  15. The RNAi pathway plays a small part in Wolbachia-mediated blocking of dengue virus in mosquito cells.

    PubMed

    Terradas, Gerard; Joubert, D Albert; McGraw, Elizabeth A

    2017-03-06

    Wolbachia pipientis is an insect endosymbiont known to limit the replication of viruses including dengue and Zika in their primary mosquito vector, Aedes aegypti. Wolbachia is being released into mosquito populations globally in a bid to control the diseases caused by these viruses. It is theorized that Wolbachia's priming of the insect immune system may confer protection against subsequent viral infection. Other hypotheses posit a role for competition between Wolbachia and viruses for host cellular resources. Using an A. aegypti cell line infected with Wolbachia, we tested the effects of targeting siRNAs against the major innate immune pathways on dengue virus loads. We show that while Wolbachia infection induces genes in the Toll, JAK/STAT and RNAi pathways, only reduced expression of RNAi leads to a rebound of dengue virus loads in Wolbachia-infected cells. The magnitude of the effect explained less than 10% of the total DENV load, demonstrating that blocking must be dependent on other factors in addition to the expression of RNAi. The findings bode well for the long-term stability of blocking given that immunity gene expression would likely be highly plastic and susceptible to rapid evolution.

  16. Hypervariable prophage WO sequences describe an unexpected high number of Wolbachia variants in the mosquito Culex pipiens

    PubMed Central

    Duron, Olivier; Fort, Philippe; Weill, Mylène

    2005-01-01

    Wolbachia are maternally inherited endosymbiotic bacteria that infect many arthropod species and may induce cytoplasmic incompatibility (CI) resulting in abortive embryonic development. Among all the described host species, mosquitoes of the Culex pipiens complex display the highest variability of CI crossing types. Paradoxically, searches for polymorphism in Wolbachia infecting strains and field populations hitherto failed or produced very few markers. Here, we show that an abundant source of the long-sought polymorphism lies in WO prophage sequences present in multiple copies dispersed in the genome of Wolbachia infecting C. pipiens (wPip). We identified up to 66 different Wolbachia variants in C. pipiens strains and field populations and no occurrence of superinfection was observed. At least 49 different Wolbachia occurred in Southern Europe C. pipiens populations, and up to 10 different Wolbachia were even detected in a single population. This is in sharp contrast with North African and Cretan samples, which exhibited only six variants. The WO polymorphism appeared stable over time, and was exclusively transferred maternally. Interestingly, we found that the CI pattern previously described correlates with the variability of Gp15, a prophage protein similar to a bacterial virulence protein. WO prophage sequences thus represent variable markers that now open routes for approaching the molecular basis of CI, the host effects, the structure and dynamics of Wolbachia populations. PMID:16615218

  17. How do hosts react to endosymbionts? A new insight into the molecular mechanisms underlying the Wolbachia-host association.

    PubMed

    Zhang, Y-K; Ding, X-L; Rong, X; Hong, X-Y

    2015-02-01

    Wolbachia is an intracellular bacterium that has aroused intense interest because of its ability to alter the biology of its host in diverse ways. In the two-spotted spider mite, Tetranychus urticae, Wolbachia can induce complex cytoplasmic incompatibility (CI) phenotypes and fitness changes, although little is known about the mechanisms. In the present study, we selected a strain of T. urticae, in which Wolbachia infection was associated with strong CI and enhanced female fecundity, to investigate changes in the transcriptome of T. urticae in Wolbachia-infected vs. uninfected lines. The responses were found to be sex-specific, with the transcription of 251 genes being affected in females and 171 genes being affected in males. Some of the more profoundly affected genes in both sexes were lipocalin genes and genes involved in oxidation reduction, digestion and detoxification. Several of the differentially expressed genes have potential roles in reproduction. Interestingly, unlike certain Wolbachia transinfections in novel hosts, the Wolbachia-host association in the present study showed no clear evidence of host immune priming by Wolbachia, although a few potential immune genes were affected.

  18. Relations of Wolbachia Infection with Phylogeography of Philaenus spumarius (Hemiptera: Aphrophoridae) Populations Within and Beyond the Carpathian Contact Zone.

    PubMed

    Lis, Agata; Maryańska-Nadachowska, Anna; Kajtoch, Łukasz

    2015-08-01

    Wolbachia is the most widespread intracellular α-proteobacteria maternally inherited endosymbiont of insects and nematodes. These bacteria are associated with a number of different reproductive phenotypes of their hosts. Relatively few studies have dealt with distribution of infections across populations and with the influence of these bacteria on host genetic diversification and speciation. The aims of this study are to determine the distribution and rate of infection and to characterize the Wolbachia strains associated with Philaenus spumarius spittlebug (Hemiptera) by using multilocus sequencing typing (MLST) analysis and host phylogeography. The results showed that infection rate was significantly different between members of both main mitochondrial phylogenetic lineages of P. spumarius. We detected much higher infection rates of Wolbachia in P. spumarius populations from the north-east clade than the south-west clade. Moreover, the frequency of these infections varied within and outside the contact zone known from the Carpathians. Given the reproductive alterations which are often associated with this endosymbiont, Wolbachia probably maintain genetic differentiation of its hosts in its contact zone in the Carpathians. This is one of the first studies demonstrating the presence of Wolbachia across a large part of the range of insect species, including the contact zone. The spread of Wolbachia in P. spumarius populations can potentially cause speciation by compromising the potential reproductive barrier between infected and uninfected populations. We discuss possible implications of Wolbachia infection inducing cytoplasmic incompatibility in the population dynamics of this spittlebug but confirm that more studies are also required.

  19. Geography has a greater effect than Wolbachia infection on population genetic structure in the spider mite, Tetranychus pueraricola.

    PubMed

    Chen, Y-T; Zhang, Y-K; Du, W-X; Jin, P-Y; Hong, X-Y

    2016-10-01

    Wolbachia is an intracellular symbiotic bacterium that infects various spider mite species and is associated with alterations in host reproduction, which indicates the potential role in mite evolution. However, studies of Wolbachia infections in the spider mite Tetranychus pueraricola, a major agricultural pest, are limited. Here, we used multilocus sequence typing to determine Wolbachia infection status and examined the relationship between Wolbachia infection status and mitochondrial diversity in T. pueraricola from 12 populations in China. The prevalence of Wolbachia ranged from 2.8 to 50%, and three strains (wTpue1, wTpue2, and wTpue3) were identified. We also found double infections (wTpue1 + wTpue3) within the same individuals. Furthermore, the wTpue1 strain caused weak cytoplasmic incompatibility (CI) (egg hatchability ~55%), whereas another widespread strain, wTpue3, did not induce CI. There was no reduction in mitochondrial DNA (mtDNA) or nuclear DNA diversity among infected individuals, and mtDNA haplotypes did not correspond to specific Wolbachia strains. Phylogenetic analysis and analysis of molecular variance revealed that the distribution of mtDNA and nuclear DNA haplotypes were significantly associated with geography. These findings indicate that Wolbachia infection in T. pueraricola is complex, but T. pueraricola genetic differentiation likely resulted from substantial geographic isolation.

  20. Passenger well-being in airplanes.

    PubMed

    Hinninghofen, H; Enck, P

    2006-10-30

    Passenger well-being is influenced by cabin environmental conditions which interact with individual passenger characteristics like age and health conditions. Cabin environment is composed of different aspects, some of which have a direct influence on gastrointestinal functions and may directly generate nausea, such as cabin pressure, oxygen saturation, and motion or vibration. For example, it has been shown that available cabin pressure during normal flight altitude can significantly inhibit gastric emptying and induce dyspepsia-like symptoms when associated with a fibre-rich meal. Other aspects of the cabin environment such as space and variability of seating, air quality, and noise, also have been shown to modulate (reduce or increase) discomfort and nausea during flights. Individual passenger characteristics and health status also have been demonstrated to increase vulnerability to adverse health outcomes and discomfort.

  1. Locating influential nodes via dynamics-sensitive centrality

    PubMed Central

    Liu, Jian-Guo; Lin, Jian-Hong; Guo, Qiang; Zhou, Tao

    2016-01-01

    With great theoretical and practical significance, locating influential nodes of complex networks is a promising issue. In this paper, we present a dynamics-sensitive (DS) centrality by integrating topological features and dynamical properties. The DS centrality can be directly applied in locating influential spreaders. According to the empirical results on four real networks for both susceptible-infected-recovered (SIR) and susceptible-infected (SI) spreading models, the DS centrality is more accurate than degree, k-shell index and eigenvector centrality. PMID:26905891

  2. Wolbachia uses host microRNAs to manipulate host gene expression and facilitate colonization of the dengue vector Aedes aegypti

    PubMed Central

    Hussain, Mazhar; Frentiu, Francesca D.; Moreira, Luciano A.; O'Neill, Scott L.; Asgari, Sassan

    2011-01-01

    The obligate endosymbiont Wolbachia pipientis is found in a wide range of invertebrates where they are best known for manipulating host reproduction. Recent studies have shown that Wolbachia also can modulate the lifespan of host insects and interfere with the development of human pathogens in mosquito vectors. Despite considerable study, very little is known about the molecular interactions between Wolbachia and its hosts that might mediate these effects. Using microarrays, we show that the microRNA (miRNA) profile of the mosquito, Aedes aegypti, is significantly altered by the wMelPop-CLA strain of W. pipientis. We found that a host miRNA (aae-miR-2940) is induced after Wolbachia infection in both mosquitoes and cell lines. One target of aae-miR-2940 is the Ae. aegypti metalloprotease gene. Interestingly, expression of the target gene was induced after Wolbachia infection, ectopic expression of the miRNA independent of Wolbachia, or transfection of an artificial mimic of the miRNA into mosquito cells. We also confirmed the interaction of aae-miR-2940 with the target sequences using GFP as a reporter gene. Silencing of the metalloprotease gene in both Wolbachia-infected cells and adult mosquitoes led to a significant reduction in Wolbachia density, as did inhibition of the miRNA in cells. These results indicate that manipulation of the mosquito metalloprotease gene via aae-miR-2940 is crucial for efficient maintenance of the endosymbiont. This report shows how Wolbachia alters the host miRNA profile and provides insight into the mechanisms of host manipulation used by this widespread endosymbiont. PMID:21576469

  3. Passenger ride quality in transport aircraft

    NASA Technical Reports Server (NTRS)

    Jacobson, I. D.; Kuhlthau, A. R.; Richards, L. G.; Conner, D. W.

    1978-01-01

    Quantitative relationships are presented which can be used to account for passenger ride quality in transport aircraft. These relations can be used to predict passenger comfort and satisfaction under a variety of flight conditions. Several applications are detailed, including evaluation of use of spoilers to attenuate trailing vortices, identifying key elements in a complex maneuver which leads to discomfort, determining noise/motion tradeoffs, evaluating changes in wing loading, and others. Variables included in the models presented are motion, noise, temperature, pressure, and seating.

  4. 5. RAILROAD TRRACKS LEADING TO PAINT & REPAIR SHOP; PASSENGER ...

    Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

    5. RAILROAD TRRACKS LEADING TO PAINT & REPAIR SHOP; PASSENGER CAR SHOP TO THE LEFT - Baltimore & Ohio Railroad, Mount Clare Passenger Car Shop, Southwest corner of Pratt & Poppleton Streets, Baltimore, Independent City, MD

  5. 14. AERIAL VIEW OF ENGINE DISPLAY INSIDE PASSENGER CAR SHOP ...

    Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

    14. AERIAL VIEW OF ENGINE DISPLAY INSIDE PASSENGER CAR SHOP (NOW A TRANSPORTATION MUSEUM) - Baltimore & Ohio Railroad, Mount Clare Passenger Car Shop, Southwest corner of Pratt & Poppleton Streets, Baltimore, Independent City, MD

  6. 6. GENERAL VIEW OF CUPOLA AND SECOND FLOOR OF PASSENGER ...

    Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

    6. GENERAL VIEW OF CUPOLA AND SECOND FLOOR OF PASSENGER CAR SHOP - Baltimore & Ohio Railroad, Mount Clare Passenger Car Shop, Southwest corner of Pratt & Poppleton Streets, Baltimore, Independent City, MD

  7. 3. GENERAL VIEW OF PASSENGER CAR SHOP; RAILROAD TRACKS IN ...

    Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

    3. GENERAL VIEW OF PASSENGER CAR SHOP; RAILROAD TRACKS IN FOREGROUND - Baltimore & Ohio Railroad, Mount Clare Passenger Car Shop, Southwest corner of Pratt & Poppleton Streets, Baltimore, Independent City, MD

  8. Teen Drivers’ Perceptions of Their Peer Passengers: Qualitative Study

    PubMed Central

    Haynie, Denise L.; Luthers, Christina; Perlus, Jessamyn; Gerber, Eli; Ouimet, Marie Claude; Klauer, Sheila G.; Simons-Morton, Bruce

    2016-01-01

    Background The presence of peer passengers increases teenage drivers’ fatal crash risk. Distraction and social influence are the two main factors that have been associated with increased risk. Teen drivers’ perceptions of their peer passengers on these factors could inform our understanding of the conditions under which peer passengers increase crash risk or promote safer driving. The purpose of this study was to examine teen drivers’ perceptions of their peer passengers on distraction and social influence. Method A convenience sample of male and female drivers participated in a semi-structured interview that included questions on their perceptions of the effects of peer passengers on driving on distraction and social influence. The analysis of the interviews was guided by a grounded theory approach. Findings Teenage drivers were aware of the risk that peer passengers posed. Some described having passengers in the vehicle as distracting, and recognized that the level of distraction increased with the number of passengers in the vehicle. Drivers that felt responsible for the safety of their peer passengers described strategies they used to control the in-vehicle environment. Drivers described driving with passengers as a performance, and articulated direct and indirect sources of pressure, gender norms, and unspoken expectations of their passengers as influencing their driving behavior. Conclusions The influence of passengers is situation specific and dependent on whom the passenger(s) may be. Passenger influence may be either protective or harmful, depending on the circumstances. Some passengers exert direct influence, but often their influence appears more indirect and subtle. PMID:27346920

  9. 46 CFR 122.506 - Passenger safety orientation.

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    ... 46 Shipping 4 2014-10-01 2014-10-01 false Passenger safety orientation. 122.506 Section 122.506... Preparations for Emergencies § 122.506 Passenger safety orientation. (a) Except as allowed by paragraphs (b... safety information. (e) On a vessel on a voyage of more than 24 hours duration, passengers shall...

  10. 19 CFR 4.80a - Coastwise transportation of passengers.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 19 Customs Duties 1 2010-04-01 2010-04-01 false Coastwise transportation of passengers. 4.80a... transportation of passengers. (a) For the purposes of this section, the following terms will have the meaning set... to the prohibition in this section is the transportation of passengers between ports in Puerto...

  11. 14 CFR 19-7 - Passenger origin-destination survey.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... 14 Aeronautics and Space 4 2012-01-01 2012-01-01 false Passenger origin-destination survey. Sec... AIR CARRIERS Operating Statistics Classifications Sec. 19-7 Passenger origin-destination survey. (a... carriers) shall participate in a Passenger Origin-Destination (O & D) Survey covering domestic...

  12. 49 CFR 177.870 - Regulations for passenger carrying vehicles.

    Code of Federal Regulations, 2012 CFR

    2012-10-01

    ... PUBLIC HIGHWAY Regulations Applying to Hazardous Material on Motor Vehicles Carrying Passengers for Hire... apply to vehicles transporting passengers and property. (b) No Class 1 (explosive) materials or other hazardous materials on passenger-carrying vehicles, exceptions. No hazardous materials except...

  13. 49 CFR 239.103 - Passenger train emergency simulations.

    Code of Federal Regulations, 2012 CFR

    2012-10-01

    ... passenger train service that results in serious injury or death to one or more persons and property damage... 49 Transportation 4 2012-10-01 2012-10-01 false Passenger train emergency simulations. 239.103... ADMINISTRATION, DEPARTMENT OF TRANSPORTATION PASSENGER TRAIN EMERGENCY PREPAREDNESS Specific Requirements §...

  14. 49 CFR 239.103 - Passenger train emergency simulations.

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ... passenger train service that results in serious injury or death to one or more persons and property damage... 49 Transportation 4 2011-10-01 2011-10-01 false Passenger train emergency simulations. 239.103... ADMINISTRATION, DEPARTMENT OF TRANSPORTATION PASSENGER TRAIN EMERGENCY PREPAREDNESS Specific Requirements §...

  15. 49 CFR 383.117 - Requirements for passenger endorsement.

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ... passenger endorsement must have knowledge covering the following topics: (1) Proper procedures for loading/unloading passengers; (2) Proper use of emergency exits, including push-out windows; (3) Proper responses to such emergency situations as fires and unruly passengers; (4) Proper procedures at...

  16. 49 CFR 383.117 - Requirements for passenger endorsement.

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ... passenger endorsement must have knowledge covering the following topics: (1) Proper procedures for loading/unloading passengers; (2) Proper use of emergency exits, including push-out windows; (3) Proper responses to such emergency situations as fires and unruly passengers; (4) Proper procedures at...

  17. 49 CFR 374.315 - Transportation of passengers with disabilities.

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ... 49 Transportation 5 2010-10-01 2010-10-01 false Transportation of passengers with disabilities....315 Transportation of passengers with disabilities. Service provided by a carrier to passengers with disabilities is governed by the provisions of 42 U.S.C. 12101 et seq., and regulations promulgated...

  18. Older Driver and Passenger Collaboration for Wayfinding in Unfamiliar Areas

    ERIC Educational Resources Information Center

    Bryden, Kelly Jane; Charlton, Judith; Oxley, Jennifer; Lowndes, Georgia

    2014-01-01

    Passenger collaboration offers a potential compensatory strategy to assist older drivers who have difficulty driving in unfamiliar areas (wayfinding). This article describes a survey of 194 healthy, community-dwelling older drivers and their regular passengers to investigate how passengers assist drivers, and to identify the characteristics of…

  19. 46 CFR 115.910 - Passenger Ship Safety Certificate.

    Code of Federal Regulations, 2012 CFR

    2012-10-01

    ... 46 Shipping 4 2012-10-01 2012-10-01 false Passenger Ship Safety Certificate. 115.910 Section 115... Ship Safety Certificate. (a) A vessel that carries more than 12 passengers on an international voyage must have a valid SOLAS Passenger Ship Safety Certificate. The Commandant authorizes the...

  20. 46 CFR 115.910 - Passenger Ship Safety Certificate.

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ... 46 Shipping 4 2010-10-01 2010-10-01 false Passenger Ship Safety Certificate. 115.910 Section 115... Ship Safety Certificate. (a) A vessel that carries more than 12 passengers on an international voyage must have a valid SOLAS Passenger Ship Safety Certificate. The Commandant issues the original...

  1. 46 CFR 115.910 - Passenger Ship Safety Certificate.

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    ... 46 Shipping 4 2014-10-01 2014-10-01 false Passenger Ship Safety Certificate. 115.910 Section 115... Ship Safety Certificate. (a) A vessel that carries more than 12 passengers on an international voyage must have a valid SOLAS Passenger Ship Safety Certificate. The Commandant authorizes the...

  2. 46 CFR 71.75-5 - Passenger Ship Safety Certificate.

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    ... 46 Shipping 3 2014-10-01 2014-10-01 false Passenger Ship Safety Certificate. 71.75-5 Section 71.75... Passenger Ship Safety Certificate. (a) All vessels on or certificated for an international voyage are required to have a “ SOLAS Passenger Ship Safety Certificate.” (b) All such vessels shall meet...

  3. 46 CFR 71.75-5 - Passenger Ship Safety Certificate.

    Code of Federal Regulations, 2012 CFR

    2012-10-01

    ... 46 Shipping 3 2012-10-01 2012-10-01 false Passenger Ship Safety Certificate. 71.75-5 Section 71.75... Passenger Ship Safety Certificate. (a) All vessels on or certificated for an international voyage are required to have a “ SOLAS Passenger Ship Safety Certificate.” (b) All such vessels shall meet...

  4. 46 CFR 115.910 - Passenger Ship Safety Certificate.

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ... 46 Shipping 4 2013-10-01 2013-10-01 false Passenger Ship Safety Certificate. 115.910 Section 115... Ship Safety Certificate. (a) A vessel that carries more than 12 passengers on an international voyage must have a valid SOLAS Passenger Ship Safety Certificate. The Commandant authorizes the...

  5. 46 CFR 115.910 - Passenger Ship Safety Certificate.

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ... 46 Shipping 4 2011-10-01 2011-10-01 false Passenger Ship Safety Certificate. 115.910 Section 115... Ship Safety Certificate. (a) A vessel that carries more than 12 passengers on an international voyage must have a valid SOLAS Passenger Ship Safety Certificate. The Commandant authorizes the...

  6. 46 CFR 71.75-5 - Passenger Ship Safety Certificate.

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ... 46 Shipping 3 2010-10-01 2010-10-01 false Passenger Ship Safety Certificate. 71.75-5 Section 71.75... Passenger Ship Safety Certificate. (a) All vessels on an international voyage are required to have a “Passenger Ship Safety Certificate.” (b) All such vessels shall meet the requirements of this chapter...

  7. 46 CFR 71.75-5 - Passenger Ship Safety Certificate.

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ... 46 Shipping 3 2011-10-01 2011-10-01 false Passenger Ship Safety Certificate. 71.75-5 Section 71.75... Passenger Ship Safety Certificate. (a) All vessels on or certificated for an international voyage are required to have a “ SOLAS Passenger Ship Safety Certificate.” (b) All such vessels shall meet...

  8. 46 CFR 71.75-5 - Passenger Ship Safety Certificate.

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ... 46 Shipping 3 2013-10-01 2013-10-01 false Passenger Ship Safety Certificate. 71.75-5 Section 71.75... Passenger Ship Safety Certificate. (a) All vessels on or certificated for an international voyage are required to have a “ SOLAS Passenger Ship Safety Certificate.” (b) All such vessels shall meet...

  9. 14 CFR 29.1413 - Safety belts: passenger warning device.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... 14 Aeronautics and Space 1 2011-01-01 2011-01-01 false Safety belts: passenger warning device. 29.1413 Section 29.1413 Aeronautics and Space FEDERAL AVIATION ADMINISTRATION, DEPARTMENT OF....1413 Safety belts: passenger warning device. (a) If there are means to indicate to the passengers...

  10. 14 CFR 29.1413 - Safety belts: passenger warning device.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... 14 Aeronautics and Space 1 2014-01-01 2014-01-01 false Safety belts: passenger warning device. 29.1413 Section 29.1413 Aeronautics and Space FEDERAL AVIATION ADMINISTRATION, DEPARTMENT OF....1413 Safety belts: passenger warning device. (a) If there are means to indicate to the passengers...

  11. 14 CFR 29.1413 - Safety belts: passenger warning device.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 14 Aeronautics and Space 1 2010-01-01 2010-01-01 false Safety belts: passenger warning device. 29.1413 Section 29.1413 Aeronautics and Space FEDERAL AVIATION ADMINISTRATION, DEPARTMENT OF....1413 Safety belts: passenger warning device. (a) If there are means to indicate to the passengers...

  12. 14 CFR 29.1413 - Safety belts: passenger warning device.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... 14 Aeronautics and Space 1 2012-01-01 2012-01-01 false Safety belts: passenger warning device. 29.1413 Section 29.1413 Aeronautics and Space FEDERAL AVIATION ADMINISTRATION, DEPARTMENT OF....1413 Safety belts: passenger warning device. (a) If there are means to indicate to the passengers...

  13. 14 CFR 29.1413 - Safety belts: passenger warning device.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... 14 Aeronautics and Space 1 2013-01-01 2013-01-01 false Safety belts: passenger warning device. 29.1413 Section 29.1413 Aeronautics and Space FEDERAL AVIATION ADMINISTRATION, DEPARTMENT OF....1413 Safety belts: passenger warning device. (a) If there are means to indicate to the passengers...

  14. 49 CFR 374.315 - Transportation of passengers with disabilities.

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ... 49 Transportation 5 2013-10-01 2013-10-01 false Transportation of passengers with disabilities....315 Transportation of passengers with disabilities. Service provided by a carrier to passengers with disabilities is governed by the provisions of 42 U.S.C. 12101 et seq., and regulations promulgated...

  15. 49 CFR 374.315 - Transportation of passengers with disabilities.

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    ... 49 Transportation 5 2014-10-01 2014-10-01 false Transportation of passengers with disabilities....315 Transportation of passengers with disabilities. Service provided by a carrier to passengers with disabilities is governed by the provisions of 42 U.S.C. 12101 et seq., and regulations promulgated...

  16. 49 CFR 374.315 - Transportation of passengers with disabilities.

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ... 49 Transportation 5 2011-10-01 2011-10-01 false Transportation of passengers with disabilities....315 Transportation of passengers with disabilities. Service provided by a carrier to passengers with disabilities is governed by the provisions of 42 U.S.C. 12101 et seq., and regulations promulgated...

  17. 49 CFR 374.315 - Transportation of passengers with disabilities.

    Code of Federal Regulations, 2012 CFR

    2012-10-01

    ... 49 Transportation 5 2012-10-01 2012-10-01 false Transportation of passengers with disabilities....315 Transportation of passengers with disabilities. Service provided by a carrier to passengers with disabilities is governed by the provisions of 42 U.S.C. 12101 et seq., and regulations promulgated...

  18. 49 CFR 541.5 - Requirements for passenger motor vehicles.

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ... 49 Transportation 6 2010-10-01 2010-10-01 false Requirements for passenger motor vehicles. 541.5... TRAFFIC SAFETY ADMINISTRATION, DEPARTMENT OF TRANSPORTATION FEDERAL MOTOR VEHICLE THEFT PREVENTION STANDARD § 541.5 Requirements for passenger motor vehicles. (a) Each passenger motor vehicle subject...

  19. 49 CFR 541.5 - Requirements for passenger motor vehicles.

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ... 49 Transportation 6 2011-10-01 2011-10-01 false Requirements for passenger motor vehicles. 541.5... TRAFFIC SAFETY ADMINISTRATION, DEPARTMENT OF TRANSPORTATION FEDERAL MOTOR VEHICLE THEFT PREVENTION STANDARD § 541.5 Requirements for passenger motor vehicles. (a) Each passenger motor vehicle subject...

  20. 46 CFR 185.502 - Crew and passenger list.

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ... 46 Shipping 7 2010-10-01 2010-10-01 false Crew and passenger list. 185.502 Section 185.502... TONS) OPERATIONS Preparations for Emergencies § 185.502 Crew and passenger list. (a) The owner.... (b) The master of a vessel required to prepare a crew and passenger list by paragraph (a) of...

  1. Identifying the Educationally Influential Physician: A Systematic Review of Approaches

    ERIC Educational Resources Information Center

    Kronberger, Matthew P.; Bakken, Lori L.

    2011-01-01

    Introduction: Previous studies have indicated that educationally influential physicians' (EIPs) interactions with peers can lead to practice changes and improved patient outcomes. However, multiple approaches have been used to identify and investigate EIPs' informal or formal influence on practice, which creates study outcomes that are difficult…

  2. Finding Influential Users in Social Media Using Association Rule Learning

    NASA Astrophysics Data System (ADS)

    Erlandsson, Fredrik; Bródka, Piotr; Borg, Anton; Johnson, Henric

    2016-04-01

    Influential users play an important role in online social networks since users tend to have an impact on one other. Therefore, the proposed work analyzes users and their behavior in order to identify influential users and predict user participation. Normally, the success of a social media site is dependent on the activity level of the participating users. For both online social networking sites and individual users, it is of interest to find out if a topic will be interesting or not. In this article, we propose association learning to detect relationships between users. In order to verify the findings, several experiments were executed based on social network analysis, in which the most influential users identified from association rule learning were compared to the results from Degree Centrality and Page Rank Centrality. The results clearly indicate that it is possible to identify the most influential users using association rule learning. In addition, the results also indicate a lower execution time compared to state-of-the-art methods.

  3. Who Are the Most Influential Emergency Physicians on Twitter?

    PubMed Central

    Riddell, Jeff; Brown, Alisha; Kovic, Ivor; Jauregui, Joshua

    2017-01-01

    Introduction Twitter has recently gained popularity in emergency medicine (EM). Opinion leaders on Twitter have significant influence on the conversation and content, yet little is known about these opinion leaders. We aimed to describe a methodology to identify the most influential emergency physicians (EP) on Twitter and present a current list. Methods We analyzed 2,234 English-language EPs on Twitter from a previously published list of Twitter accounts generated by a snowball sampling technique. Using NodeXL software, we performed a network analysis of these EPs and ranked them on three measures of influence: in-degree centrality, eigenvector centrality, and betweenness centrality. We analyzed the top 100 users in each of these three measures of influence and compiled a list of users found in the top 100 in all three measures. Results Of the 300 total users identified by one of the measures of influence, there were 142 unique users. Of the 142 unique users, 61 users were in the top 100 on all three measures of influence. We identify these 61 users as the most influential EM Twitter users. Conclusion We both describe a method for identifying the most influential users and provide a list of the 61 most influential EPs on Twitter as of January 1, 2016. This application of network science to the EM Twitter community can guide future research to better understand the networked global community of EM. PMID:28210365

  4. Social network analysis in identifying influential webloggers: A preliminary study

    NASA Astrophysics Data System (ADS)

    Hasmuni, Noraini; Sulaiman, Nor Intan Saniah; Zaibidi, Nerda Zura

    2014-12-01

    In recent years, second generation of internet-based services such as weblog has become an effective communication tool to publish information on the Web. Weblogs have unique characteristics that deserve users' attention. Some of webloggers have seen weblogs as appropriate medium to initiate and expand business. These webloggers or also known as direct profit-oriented webloggers (DPOWs) communicate and share knowledge with each other through social interaction. However, survivability is the main issue among DPOW. Frequent communication with influential webloggers is one of the way to keep survive as DPOW. This paper aims to understand the network structure and identify influential webloggers within the network. Proper understanding of the network structure can assist us in knowing how the information is exchanged among members and enhance survivability among DPOW. 30 DPOW were involved in this study. Degree centrality and betweenness centrality measurement in Social Network Analysis (SNA) were used to examine the strength relation and identify influential webloggers within the network. Thus, webloggers with the highest value of these measurements are considered as the most influential webloggers in the network.

  5. A new measure of identifying influential nodes: Efficiency centrality

    NASA Astrophysics Data System (ADS)

    Wang, Shasha; Du, Yuxian; Deng, Yong

    2017-06-01

    Identifying the influential nodes in complex networks is a fundamental and practical topic at the moment. Local metric like degree centrality measure is relatively simple and of less effectiveness, although the global metrics such as closeness and betweenness centrality measure can better identify influential nodes, still, there are some disadvantages and limitations. In this paper, a new efficiency centrality (EffC) to rank the spreaders in the whole network is proposed, which identify influential nodes by removing each node and meanwhile considering the changing degree of the whole network efficiency after removal. To evaluate the performance of our method, Susceptible-Infected-Recovered (SIR) model is used to simulate the epidemic spreading in four real networks. The experimental and simulated results show the efficiency and practicability of the proposed method. Thus, it is significant to rank spreaders in complex networks by using Network Efficiency. And our proposed EffC is proved to be a feasible and effective measure to identify influential nodes.

  6. 78 FR 39649 - Passenger Vessels Accessibility Guidelines

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-07-02

    ... From the Federal Register Online via the Government Publishing Office ARCHITECTURAL AND TRANSPORTATION BARRIERS COMPLIANCE BOARD 36 CFR Part 1196 RIN 3014-AA11 Passenger Vessels Accessibility Guidelines Correction In proposed rule document 2013-14367, appearing on pages 38102- 38159 in the issue...

  7. Techniques for Forecasting Air Passenger Traffic

    NASA Technical Reports Server (NTRS)

    Taneja, N.

    1972-01-01

    The basic techniques of forecasting the air passenger traffic are outlined. These techniques can be broadly classified into four categories: judgmental, time-series analysis, market analysis and analytical. The differences between these methods exist, in part, due to the degree of formalization of the forecasting procedure. Emphasis is placed on describing the analytical method.

  8. Evolutionary genomics place the origin of Wolbachia in nematodes, not arthropods

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Wolbachia, the most widely studied endosymbiont in arthropods, is a target for biological control of mosquito-borne diseases (malaria and dengue virus), and antibiotic elimination of infectious filarial nematodes. We sequenced and analyzed the genome of a new strain (wPpe) in the plant-parasitic nem...

  9. Double trouble: combined action of meiotic drive and Wolbachia feminization in Eurema butterflies.

    PubMed

    Kern, Peter; Cook, James M; Kageyama, Daisuke; Riegler, Markus

    2015-05-01

    Arthropod sex ratios can be manipulated by a diverse range of selfish genetic elements, including maternally inherited Wolbachia bacteria. Feminization by Wolbachia is rare but has been described for Eurema mandarina butterflies. In this species, some phenotypic and functional females, thought to be ZZ genetic males, are infected with a feminizing Wolbachia strain, wFem. Meanwhile, heterogametic WZ females are not infected with wFem. Here, we establish a quantitative PCR assay allowing reliable sexing in three Eurema species. Against expectation, all E. mandarina females, including wFem females, had only one Z chromosome that was paternally inherited. Observation of somatic interphase nuclei confirmed that W chromatin was absent in wFem females, but present in females without wFem. We conclude that the sex bias in wFem lines is due to meiotic drive (MD) that excludes the maternal Z and thus prevents formation of ZZ males. Furthermore, wFem lines may have lost the W chromosome or harbour a dysfunctional version, yet rely on wFem for female development; removal of wFem results in all-male offspring. This is the first study that demonstrates an interaction between MD and Wolbachia feminization, and it highlights endosymbionts as potentially confounding factors in MD of sex chromosomes.

  10. Female Adult Aedes albopictus Suppression by Wolbachia-Infected Male Mosquitoes

    PubMed Central

    Mains, James W.; Brelsfoard, Corey L.; Rose, Robert I.; Dobson, Stephen L.

    2016-01-01

    Dengue, chikungunya and zika viruses are pathogens with an increasing global impact. In the absence of an approved vaccine or therapy, their management relies on controlling the mosquito vectors. But traditional controls are inadequate, and the range of invasive species such as Aedes albopictus (Asian Tiger Mosquito) is expanding. Genetically modified mosquitoes are being tested, but their use has encountered regulatory barriers and public opposition in some countries. Wolbachia bacteria can cause a form of conditional sterility, which can provide an alternative to genetic modification or irradiation. It is unknown however, whether openly released, artificially infected male Ae. albopictus can competitively mate and sterilize females at a level adequate to suppress a field population. Also, the unintended establishment of Wolbachia at the introduction site could result from horizontal transmission or inadvertent female release. In 2014, an Experimental Use Permit from the United States Environmental Protection Agency approved a pilot field trial in Lexington, Kentucky, USA. Here, we present data showing localized reduction of both egg hatch and adult female numbers. The artificial Wolbachia type was not observed to establish in the field. The results are discussed in relation to the applied use of Wolbachia-infected males as a biopesticide to suppress field populations of Ae. albopictus. PMID:27659038

  11. Time to Empower Release of Insects Carrying a Dominant Lethal and Wolbachia Against Zika.

    PubMed

    Dickens, Borame L; Yang, Jie; Cook, Alex R; Carrasco, Luis R

    2016-04-01

    RIDL (release of insects with dominant lethality) and Wolbachia are 2 potentially powerful tools in the fight against Zika, but their technological advancement is being hampered by policy barriers. In this study, we discuss what could be done to overcome these regulatory deadlocks.

  12. An update on the diversity of Wolbachia in Spalangia spp. (Hymenoptera: Pteromalidae)

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Infections of Wolbachia bacteria have the potential to improve the efficacy of their host insects as biological control agents. Results of an earlier study documented numerous cases of such infections in a beneficial guild of wasps (Hymenoptera: Pteromalidae) parasitic on pest flies affecting lives...

  13. [How to fight parasitic infectious diseases with bacteria. The case of Wolbachia pipientis].

    PubMed

    March-Rosselló, Gabriel Alberto; Eiros-Bouza, José María

    2014-01-01

    In Nature, no individual can live in isolation; hence, living organisms are forced to interact with each other. This necessity has led many organisms to establish heterogeneous relations to enhance their ability to adapt to the environment, thus acquiring evolutionary advantages. These relationships are sometimes so intense, that on the long term the organisms may lose their individual identity. An example of these associations is the endosymbiotic ones, where eukaryote organisms generally harbor different prokaryote organisms. The endosymbiotic bacterium Wolbachia pipientis is a species described by Hertig and Wolbach in 1924. This microorganism can be isolated in a large variety of eukaryote organisms, with which it maintains different links. Until now, this species has only been described with 11 serogroups numbered from A to K within the Wolbachia genus. This work is intended to illustrate the relationship of Wolbachia pipientis with human pathogenic filaria and with arthropods, as well as to describe the implications of this bacterium in the treatment of filariasis. Finally, this work tries to describe recent studies that have targeted the use of artificially-created Wolbachia pipientis virulent strains that, once inoculated in infectious diseases-transmitting vectors, develop negative effects within them in order to, in this way, erradicate mosquito-transmitted infectious diseases for which no treatment is available at the moment or the prevention of its transmissibility has not been achieved.

  14. Time to Empower Release of Insects Carrying a Dominant Lethal and Wolbachia Against Zika

    PubMed Central

    Dickens, Borame L.; Yang, Jie; Cook, Alex R.; Carrasco, Luis R.

    2016-01-01

    RIDL (release of insects with dominant lethality) and Wolbachia are 2 potentially powerful tools in the fight against Zika, but their technological advancement is being hampered by policy barriers. In this study, we discuss what could be done to overcome these regulatory deadlocks. PMID:27419175

  15. Intensity of Mutualism Breakdown Is Determined by Temperature Not Amplification of Wolbachia Genes

    PubMed Central

    Frentiu, Francesca D.; Horn, Emilie; Ritchie, Fiona K.; van Swinderen, Bruno; Weible, Michael W.; O’Neill, Scott L.; Brownlie, Jeremy C.

    2016-01-01

    Wolbachia are maternally transmitted intracellular bacterial symbionts that infect approximately 40% of all insect species. Though several strains of Wolbachia naturally infect Drosophila melanogaster and provide resistance against viral pathogens, or provision metabolites during periods of nutritional stress, one virulent strain, wMelPop, reduces fly lifespan by half, possibly as a consequence of over-replication. While the mechanisms that allow wMelPop to over-replicate are still of debate, a unique tandem repeat locus in the wMelPop genome that contains eight genes, referred to as the “Octomom” locus has been identified and is thought to play an important regulatory role. Estimates of Octomom locus copy number correlated increasing copy number to both Wolbachia bacterial density and increased pathology. Here we demonstrate that infected fly pathology is not dependent on an increased Octomom copy number, but does strongly correlate with increasing temperature. When measured across developmental time, we also show Octomom copy number to be highly variable across developmental time within a single generation. Using a second pathogenic strain of Wolbachia, we further demonstrate reduced insect lifespan can occur independently of a high Octomom locus copy number. Taken together, this data demonstrates that the mechanism/s of wMelPop virulence is more complex than has been previously described. PMID:27661080

  16. Novel estimates of Aedes aegypti (Diptera: Culicidae) population size and adult survival based on Wolbachia releases.

    PubMed

    Ritchie, Scott A; Montgomery, Brian L; Hoffmann, Ary A

    2013-05-01

    The size of Aedes aegypti (L.) mosquito populations and adult survival rates have proven difficult to estimate because of a lack of consistent quantitative measures to equate sampling methods, such as adult trapping, to actual population size. However, such estimates are critical for devising control methods and for modeling the transmission of dengue and other infectious agents carried by this species. Here we take advantage of recent releases of Wolbachia-infected Ae. aegypti coupled with the results of ongoing monitoring to estimate the size of adult Ae. aegypti populations around Cairns in far north Queensland, Australia. Based on the association between released adults infected with Wolbachia and data from Biogents Sentinel traps, we show that data from two locations are consistent with population estimates of approximately 5-10 females per house and daily survival rates of 0.7-0.9 for the released Wolbachia-infected females. Moreover, we estimate that networks of Biogents Sentinel traps at a density of one per 15 houses capture around 5-10% of the adult population per week, and provide a rapid estimate of the absolute population size of Ae. aegypti. These data are discussed with respect to release rates and monitoring in future Wolbachia releases and also the levels of suppression required to reduce dengue transmission.

  17. Wolbachia MLST marker diversity from multiply infected northern corn rootworm (Diabrotica barberi) populations

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The northern corn rootworm (NCR, Diabrotica barberi) in eastern and central North America exhibits at least three distinct populations with respect to Wolbachia infection: uninfected; singly infected; multiply infected. The infected states are associated with different mtDNA haplotypes and reduced m...

  18. Double trouble: combined action of meiotic drive and Wolbachia feminization in Eurema butterflies

    PubMed Central

    Kern, Peter; Cook, James M.; Kageyama, Daisuke; Riegler, Markus

    2015-01-01

    Arthropod sex ratios can be manipulated by a diverse range of selfish genetic elements, including maternally inherited Wolbachia bacteria. Feminization by Wolbachia is rare but has been described for Eurema mandarina butterflies. In this species, some phenotypic and functional females, thought to be ZZ genetic males, are infected with a feminizing Wolbachia strain, wFem. Meanwhile, heterogametic WZ females are not infected with wFem. Here, we establish a quantitative PCR assay allowing reliable sexing in three Eurema species. Against expectation, all E. mandarina females, including wFem females, had only one Z chromosome that was paternally inherited. Observation of somatic interphase nuclei confirmed that W chromatin was absent in wFem females, but present in females without wFem. We conclude that the sex bias in wFem lines is due to meiotic drive (MD) that excludes the maternal Z and thus prevents formation of ZZ males. Furthermore, wFem lines may have lost the W chromosome or harbour a dysfunctional version, yet rely on wFem for female development; removal of wFem results in all-male offspring. This is the first study that demonstrates an interaction between MD and Wolbachia feminization, and it highlights endosymbionts as potentially confounding factors in MD of sex chromosomes. PMID:25948567

  19. Wolbachia as an “Infectious” Extrinsic Factor Manipulating Host Signaling Pathways

    PubMed Central

    Negri, Ilaria

    2011-01-01

    Wolbachia pipientis is a widespread endosymbiont of filarial nematodes and arthropods. While in worms the symbiosis is obligate, in arthropods Wolbachia induces several reproductive manipulations (i.e., cytoplasmic incompatibility, parthenogenesis, feminization of genetic males, and male-killing) in order to increase the number of infected females. These various phenotypic effects may be linked to differences in host physiology, and in particular to endocrine-related processes governing growth, development, and reproduction. Indeed, a number of evidences links Wolbachia symbiosis to insulin and ecdysteroid signaling, two multilayered pathways known to work antagonistically, jointly or even independently for the regulation of different molecular networks. At present it is not clear whether Wolbachia manipulates one pathway, thus affecting other related metabolic networks, or if it targets both pathways, even interacting at several points in each of them. Interestingly, in view of the interplay between hormone signaling and epigenetic machinery, a direct influence of the “infection” on hormonal signaling involving ecdysteroids might be achievable through the manipulation of the host’s epigenetic pathways. PMID:22654845

  20. Wolbachia Occurrence in Planthopper (Hemiptera: Delphacidae) Vectors of Cereal Viruses in Argentina.

    PubMed

    Mattio, M F; Argüello Caro, E B; Rodriguero, M S; Dumón, A D; Alemandri, V M; Truol, G

    2015-08-01

    Maize (Zea mays L.) and wheat (Triticum aestivum L.) are the most important cereal crops for the Argentinean economy and are affected by several diseases. Different planthopper species transmit causal agents of some of those diseases, including Mal de Río Cuarto virus, barley yellow striate mosaic virus, and the recently proposed maize yellow striate virus. Many planthopper species are sap feeders and therefore are expected to host bacteria providing essential nutrients lacking in the diet. Previous studies have evidenced that some of these bacterial symbionts are involved in the virus transmission. Wolbachia is a group of obligate intracellular bacteria infecting numerous arthropod species and causing reproductive alterations in their hosts. These bacteria have been detected in planthopper species, considered rice pests in various regions of the world. To date, Wolbachia infection status of planthopper species of Argentina is unknown. Amplification by PCR and sequencing of 16S rDNA, wsp- and ftsZ-specific genes demonstrated Wolbachia infection in Caenodelphax teapae (Fowler), Delphacodes kuscheli Fennah, Pyrophagus tigrinus Remes Lenicov & Varela, Tagosodes orizicolus (Muir), and Toya propinqua (Fieber). This is the first report of Wolbachia in delphacid vectors of viruses affecting maize and wheat. An understanding of the bacterial diversity harbored by these insect vectors could lead to new options for future management of diseases of economically important crops in a developing country.

  1. Molecular genetics of the Wolbachia endosymbionts that infect the parasitoids of tephritid fruit flies.

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Limited information exists on the molecular genetics of the Wolbachia endosymbionts that infect the parasitoids of tephritid fruit flies. A better understanding of the bacteria could allow sex ratio manipulations that would improve the mass-rearing of natural enemies. Scientists at the Center for Me...

  2. Microsatellite and Wolbachia analysis in Rhagoletis cerasi natural populations: population structuring and multiple infections

    PubMed Central

    Augustinos, Antonios A; Asimakopoulou, Anastasia K; Moraiti, Cleopatra A; Mavragani-Tsipidou, Penelope; Papadopoulos, Nikolaos T; Bourtzis, Kostas

    2014-01-01

    Rhagoletis cerasi (Diptera: Tephritidae) is a major pest of sweet and sour cherries in Europe and parts of Asia. Despite its economic significance, there is a lack of studies on the genetic structure of R. cerasi populations. Elucidating the genetic structure of insects of economic importance is crucial for developing phenological-predictive models and environmental friendly control methods. All natural populations of R. cerasi have been found to harbor the endosymbiont Wolbachia pipientis, which widely affects multiple biological traits contributing to the evolution of its hosts, and has been suggested as a tool for the biological control of insect pests and disease vectors. In the current study, the analysis of 18 R. cerasi populations collected in Greece, Germany, and Russia using 13 microsatellite markers revealed structuring of R. cerasi natural populations, even at close geographic range. We also analyzed the Wolbachia infection status of these populations using 16S rRNA-, MLST- and wsp-based approaches. All 244 individuals screened were positive for Wolbachia. Our results suggest the fixation of the wCer1 strain in Greece while wCer2, wCer4, wCer5, and probably other uncharacterized strains were also detected in multiply infected individuals. The role of Wolbachia and its potential extended phenotypes needs a thorough investigation in R. cerasi. Our data suggest an involvement of this symbiont in the observed restriction in the gene flow in addition to a number of different ecological factors. PMID:24963388

  3. Dynamic Wolbachia prevalence in Acromyrmex leaf-cutting ants: potential for a nutritional symbiosis.

    PubMed

    Andersen, S B; Boye, M; Nash, D R; Boomsma, J J

    2012-07-01

    Wolbachia are renowned as reproductive parasites, but their phenotypic effects in eusocial insects are not well understood. We used a combination of qrt-PCR, fluorescence in situ hybridization and laser scanning confocal microscopy to evaluate the dynamics of Wolbachia infections in the leaf-cutting ant Acromyrmex octospinosus across developmental stages of sterile workers. We confirm that workers are infected with one or two widespread wsp genotypes of Wolbachia, show that colony prevalence is always 100% and characterize two rare recombinant genotypes. One dominant genotype is always present and most abundant, whereas another only proliferates in adult workers of some colonies and is barely detectable in larvae and pupae. An explanation may be that Wolbachia genotypes compete for host resources in immature stages while adult tissues provide substantially more niche space. Tissue-specific prevalence of the two genotypes differs, with the rarer genotype being over-represented in the adult foregut and thorax muscles. Both genotypes occur extracellularly in the foregut, suggesting an unknown mutualistic function in worker ant nutrition. Both genotypes are also abundant in the faecal fluid of the ants, suggesting that they may have extended functional phenotypes in the fungus garden that the ants manure with their own faeces.

  4. Constraints on the use of lifespan-shortening Wolbachia to control dengue fever.

    PubMed

    Schraiber, Joshua G; Kaczmarczyk, Angela N; Kwok, Ricky; Park, Miran; Silverstein, Rachel; Rutaganira, Florentine U; Aggarwal, Taruna; Schwemmer, Michael A; Hom, Carole L; Grosberg, Richard K; Schreiber, Sebastian J

    2012-03-21

    Dengue fever, a viral disease spread by the mosquito Aedes aegypti, affects 50-100 million people a year in many tropical countries. Because the virus must incubate within mosquito hosts for two weeks before being able to transmit the infection, shortening the lifespan of mosquitoes may curtail dengue transmission. We developed a continuous time reaction-diffusion model of the spatial spread of Wolbachia through a population of A. aegypti. This model incorporates the lifespan-shortening effects of Wolbachia on infected A. aegypti and the fitness advantage to infected females due to cytoplasmic incompatibility (CI). We found that local establishment of the Wolbachia infection can occur if the fitness advantage due to CI exceeds the fitness reduction due to lifespan-shortening effects, in accordance with earlier results concerning fecundity reduction. However, spatial spread is possible only if the fitness advantage due to CI is twice as great as the fitness reduction due to lifespan shortening effects. Moreover, lifespan-shortening and fecundity-reduction can have different effects on the speed of wave-retreat. Using data from the literature, we estimated all demographic parameters for infected and uninfected mosquitoes and computed the velocities of spread of infection. Our most optimistic estimates suggest that the spatial spread of lifespan-shortening Wolbachia may be so slow that efficient spatial spread would require a prohibitively large number of point releases. However, as these estimates of demographic parameters may not accurately reflect natural conditions, further research is necessary to corroborate these predictions.

  5. Detecting influential observations in nonlinear regression modeling of groundwater flow

    USGS Publications Warehouse

    Yager, R.M.

    1998-01-01

    Nonlinear regression is used to estimate optimal parameter values in models of groundwater flow to ensure that differences between predicted and observed heads and flows do not result from nonoptimal parameter values. Parameter estimates can be affected, however, by observations that disproportionately influence the regression, such as outliers that exert undue leverage on the objective function. Certain statistics developed for linear regression can be used to detect influential observations in nonlinear regression if the models are approximately linear. This paper discusses the application of Cook's D, which measures the effect of omitting a single observation on a set of estimated parameter values, and the statistical parameter DFBETAS, which quantifies the influence of an observation on each parameter. The influence statistics were used to (1) identify the influential observations in the calibration of a three-dimensional, groundwater flow model of a fractured-rock aquifer through nonlinear regression, and (2) quantify the effect of omitting influential observations on the set of estimated parameter values. Comparison of the spatial distribution of Cook's D with plots of model sensitivity shows that influential observations correspond to areas where the model heads are most sensitive to certain parameters, and where predicted groundwater flow rates are largest. Five of the six discharge observations were identified as influential, indicating that reliable measurements of groundwater flow rates are valuable data in model calibration. DFBETAS are computed and examined for an alternative model of the aquifer system to identify a parameterization error in the model design that resulted in overestimation of the effect of anisotropy on horizontal hydraulic conductivity.

  6. Genetic and functional characterization of the type IV secretion system in Wolbachia.

    PubMed

    Rancès, Edwige; Voronin, Denis; Tran-Van, Van; Mavingui, Patrick

    2008-07-01

    A type IV secretion system (T4SS) is used by many symbiotic and pathogenic intracellular bacteria for the successful infection of and survival, proliferation, and persistence within hosts. In this study, the presence and function of the T4SS in Wolbachia strains were investigated by a combination of genetic screening and immunofluorescence microscopy. Two operons of virB-virD4 loci were found in the genome of Wolbachia pipientis strain wAtab3, from the Hymenoptera Asobara tabida, and strain wRi, infecting Drosophila simulans. One operon consisted of five vir genes (virB8, virB9, virB10, virB11, and virD4) and the downstream wspB locus. The other operon was composed of three genes (virB3, virB4, and virB6) and included four additional open reading frames (orf1 to orf4) orientated in the same direction. In cell culture and insect hosts infected with different Wolbachia strains, the bona fide vir genes were polycistronically transcribed, together with the downstream adjacent loci, notably, as virB8 to virD4 and wspB and as virB3, virB4, virB6, and orf1 to orf4. Two peptides encompassing conserved C and N termini of the Wolbachia VirB6 protein were used for the production of polyclonal antibodies. Anti-VirB6 antibodies could detect the corresponding recombinant protein by chemifluorescence on Western blots of total proteins from Escherichia coli transformants and Wolbachia strains cultured in cell lines. Using immunofluorescence microscopy, we further demonstrated that the VirB6 protein was produced by Wolbachia strains in ovaries of insects harboring wAtab3 or wRi and cell lines infected with wAlbB or wMelPop. As VirB6 is known to associate with other VirB proteins to form a membrane-spanning structure, this finding suggests that a T4SS may function in Wolbachia.

  7. 75 FR 26839 - Metrics and Standards for Intercity Passenger Rail Service under Section 207 of the Passenger...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-05-12

    ... performance and service quality of intercity passenger train operations. In compliance with the statute, the FRA and Amtrak jointly drafted performance metrics and standards for intercity passenger rail service... Intercity Passenger Rail Service,'' on the FRA's Web site. Simultaneously, the FRA published a notice in...

  8. No Evidence for Wolbachia Phenotypic Effects in Newly Mated Queens of the Fire Ant Solenopsis invicta (Hymenoptera: Formicidae)

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Wolbachia are intracellular bacteria that commonly infect many arthropods and some nematodes. In arthropods, these bacteria often induce phenotypic effects that enhance their own spread within host populations, despite sometimes also having deleterious effects on host fecundity and viability. Wolbac...

  9. The wMel Strain of Wolbachia Reduces Transmission of Chikungunya Virus in Aedes aegypti

    PubMed Central

    Aliota, Matthew T.; Walker, Emma C.; Uribe Yepes, Alexander; Dario Velez, Ivan; Christensen, Bruce M.; Osorio, Jorge E.

    2016-01-01

    Background New approaches to preventing chikungunya virus (CHIKV) are needed because current methods are limited to controlling mosquito populations, and they have not prevented the invasion of this virus into new locales, nor have they been sufficient to control the virus upon arrival. A promising candidate for arbovirus control and prevention relies on the introduction of the intracellular bacterium Wolbachia into Aedes aegypti mosquitoes. This primarily has been proposed as a tool to control dengue virus (DENV) transmission; however, evidence suggests Wolbachia infections confer protection for Ae. aegypti against CHIKV. Although this approach holds much promise for limiting virus transmission, at present our understanding of the ability of CHIKV to infect, disseminate, and be transmitted by wMel-infected Ae. aegypti currently being used at Wolbachia release sites is limited. Methodology/Principal Findings Using Ae. aegypti infected with the wMel strain of Wolbachia that are being released in Medellin, Colombia, we report that these mosquitoes have reduced vector competence for CHIKV, even with extremely high viral titers in the bloodmeal. In addition, we examined the dynamics of CHIKV infection over the course of four to seven days post feeding. Wolbachia-infected mosquitoes remained non-infective over the duration of seven days, i.e., no infectious virus was detected in the saliva when exposed to bloodmeals of moderate viremia, but CHIKV-exposed, wild type mosquitoes did have viral loads in the saliva consistent with what has been reported elsewhere. Finally, the presence of wMel infection had no impact on the lifespan of mosquitoes as compared to wild type mosquitoes following CHIKV infection. Conclusions/Significance These results could have an impact on vector control strategies in areas where Ae. aegypti are transmitting both DENV and CHIKV; i.e., they argue for further exploration, both in the laboratory and the field, on the feasibility of expanding this

  10. Signs of neutralization in a redundant gene involved in homologous recombination in Wolbachia endosymbionts.

    PubMed

    Badawi, Myriam; Giraud, Isabelle; Vavre, Fabrice; Grève, Pierre; Cordaux, Richard

    2014-09-17

    Genomic reduction in bacterial endosymbionts occurs through large genomic deletions and long-term accumulation of mutations. The latter process involves successive steps including gene neutralization, pseudogenization, and gradual erosion until complete loss. Although many examples of pseudogenes at various levels of degradation have been reported, neutralization cases are scarce because of the transient nature of the process. Gene neutralization may occur due to relaxation of selection in nonessential genes, for example, those involved in redundant functions. Here, we report an example of gene neutralization in the homologous recombination (HR) pathway of Wolbachia, a bacterial endosymbiont of arthropods and nematodes. The HR pathway is often depleted in endosymbiont genomes, but it is apparently intact in some Wolbachia strains. Analysis of 12 major HR genes showed that they have been globally under strong purifying selection during the evolution of Wolbachia strains hosted by arthropods, supporting the evolutionary importance of the HR pathway for these Wolbachia genomes. However, we detected signs of recent neutralization of the ruvA gene in a subset of Wolbachia strains, which might be related to an ancestral, clade-specific amino acid change that impaired DNA-binding activity. Strikingly, RuvA is part of the RuvAB complex involved in branch migration, whose function overlaps with the RecG helicase. Although ruvA is experiencing neutralization, recG is under strong purifying selection. Thus, our high phylogenetic resolution suggests that we identified a rare example of targeted neutralization of a gene involved in a redundant function in an endosymbiont genome.

  11. The wMel Wolbachia strain blocks dengue and invades caged Aedes aegypti populations.

    PubMed

    Walker, T; Johnson, P H; Moreira, L A; Iturbe-Ormaetxe, I; Frentiu, F D; McMeniman, C J; Leong, Y S; Dong, Y; Axford, J; Kriesner, P; Lloyd, A L; Ritchie, S A; O'Neill, S L; Hoffmann, A A

    2011-08-24

    Dengue fever is the most important mosquito-borne viral disease of humans with more than 50 million cases estimated annually in more than 100 countries. Disturbingly, the geographic range of dengue is currently expanding and the severity of outbreaks is increasing. Control options for dengue are very limited and currently focus on reducing population abundance of the major mosquito vector, Aedes aegypti. These strategies are failing to reduce dengue incidence in tropical communities and there is an urgent need for effective alternatives. It has been proposed that endosymbiotic bacterial Wolbachia infections of insects might be used in novel strategies for dengue control. For example, the wMelPop-CLA Wolbachia strain reduces the lifespan of adult A. aegypti mosquitoes in stably transinfected lines. This life-shortening phenotype was predicted to reduce the potential for dengue transmission. The recent discovery that several Wolbachia infections, including wMelPop-CLA, can also directly influence the susceptibility of insects to infection with a range of insect and human pathogens has markedly changed the potential for Wolbachia infections to control human diseases. Here we describe the successful transinfection of A. aegypti with the avirulent wMel strain of Wolbachia, which induces the reproductive phenotype cytoplasmic incompatibility with minimal apparent fitness costs and high maternal transmission, providing optimal phenotypic effects for invasion. Under semi-field conditions, the wMel strain increased from an initial starting frequency of 0.65 to near fixation within a few generations, invading A. aegypti populations at an accelerated rate relative to trials with the wMelPop-CLA strain. We also show that wMel and wMelPop-CLA strains block transmission of dengue serotype 2 (DENV-2) in A. aegypti, forming the basis of a practical approach to dengue suppression.

  12. Signs of Neutralization in a Redundant Gene Involved in Homologous Recombination in Wolbachia Endosymbionts

    PubMed Central

    Badawi, Myriam; Giraud, Isabelle; Vavre, Fabrice; Grève, Pierre; Cordaux, Richard

    2014-01-01

    Genomic reduction in bacterial endosymbionts occurs through large genomic deletions and long-term accumulation of mutations. The latter process involves successive steps including gene neutralization, pseudogenization, and gradual erosion until complete loss. Although many examples of pseudogenes at various levels of degradation have been reported, neutralization cases are scarce because of the transient nature of the process. Gene neutralization may occur due to relaxation of selection in nonessential genes, for example, those involved in redundant functions. Here, we report an example of gene neutralization in the homologous recombination (HR) pathway of Wolbachia, a bacterial endosymbiont of arthropods and nematodes. The HR pathway is often depleted in endosymbiont genomes, but it is apparently intact in some Wolbachia strains. Analysis of 12 major HR genes showed that they have been globally under strong purifying selection during the evolution of Wolbachia strains hosted by arthropods, supporting the evolutionary importance of the HR pathway for these Wolbachia genomes. However, we detected signs of recent neutralization of the ruvA gene in a subset of Wolbachia strains, which might be related to an ancestral, clade-specific amino acid change that impaired DNA-binding activity. Strikingly, RuvA is part of the RuvAB complex involved in branch migration, whose function overlaps with the RecG helicase. Although ruvA is experiencing neutralization, recG is under strong purifying selection. Thus, our high phylogenetic resolution suggests that we identified a rare example of targeted neutralization of a gene involved in a redundant function in an endosymbiont genome. PMID:25230723

  13. Wolbachia Blocks Viral Genome Replication Early in Infection without a Transcriptional Response by the Endosymbiont or Host Small RNA Pathways.

    PubMed

    Rainey, Stephanie M; Martinez, Julien; McFarlane, Melanie; Juneja, Punita; Sarkies, Peter; Lulla, Aleksei; Schnettler, Esther; Varjak, Margus; Merits, Andres; Miska, Eric A; Jiggins, Francis M; Kohl, Alain

    2016-04-01

    The intracellular endosymbiotic bacterium Wolbachia can protect insects against viral infection, and is being introduced into mosquito populations in the wild to block the transmission of arboviruses that infect humans and are a major public health concern. To investigate the mechanisms underlying this antiviral protection, we have developed a new model system combining Wolbachia-infected Drosophila melanogaster cell culture with the model mosquito-borne Semliki Forest virus (SFV; Togaviridae, Alphavirus). Wolbachia provides strong antiviral protection rapidly after infection, suggesting that an early stage post-infection is being blocked. Wolbachia does appear to have major effects on events distinct from entry, assembly or exit as it inhibits the replication of an SFV replicon transfected into the cells. Furthermore, it causes a far greater reduction in the expression of proteins from the 3' open reading frame than the 5' non-structural protein open reading frame, indicating that it is blocking the replication of viral RNA. Further to this separation of the replicase proteins and viral RNA in transreplication assays shows that uncoupling of viral RNA and replicase proteins does not overcome Wolbachia's antiviral activity. This further suggests that replicative processes are disrupted, such as translation or replication, by Wolbachia infection. This may occur by Wolbachia mounting an active antiviral response, but the virus did not cause any transcriptional response by the bacterium, suggesting that this is not the case. Host microRNAs (miRNAs) have been implicated in protection, but again we found that host cell miRNA expression was unaffected by the bacterium and neither do our findings suggest any involvement of the antiviral siRNA pathway. We conclude that Wolbachia may directly interfere with early events in virus replication such as translation of incoming viral RNA or RNA transcription, and this likely involves an intrinsic (as opposed to an induced

  14. Strength of the pathogenicity caused by feminizing Wolbachia after transfer in a new host: strain or dose effect?

    PubMed

    Le Clec'h, Winka; Raimond, Maryline; Bouchon, Didier; Sicard, Mathieu

    2014-02-01

    The alphaproteobacteria Wolbachia pipientis are among the most common and widespread symbionts in the animal world. Their vertical transmission mode is predicted to favour genotypes with low virulence. On the contrary, horizontal transfers of Wolbachia from one host to another have been shown to possibly increase the symbiont virulence. This situation has been previously described when two feminizing Wolbachia strains, wVulC and wVulM, from the ovaries of the woodlouse Armadillidium vulgare were introduced into another woodlouse named Porcellio dilatatus. These two Wolbachia strains induced severe symptoms and eventually caused the death of the recipient host. However, symptoms and death appeared sooner with wVulC than with wVulM. To know whether this difference was due to variation in the dose of infection or a difference in virulence between the two Wolbachia strains, we performed controlled and gradual doses of injection with wVulC and wVulM in P. dilatatus. We showed that the two strains differed intrinsically in their virulence against P. dilatatus and that their virulence is related to the injection dose. Moreover, we showed that wVulC reached higher concentrations in the recipient host than wVulM suggesting a potential link between the bacterial titers and the levels of virulence. We also addressed the impact of the tissue source of the Wolbachia used for the transinfection and demonstrated that Wolbachia transinfected via hemolymph colonized the body of the recipient more quickly and caused accelerated symptoms compared to Wolbachia introduced via a crushed ovaries suspension.

  15. Internal spatiotemporal population dynamics of infection with three Wolbachia strains in the adzuki bean beetle, Callosobruchus chinensis (Coleoptera: Bruchidae).

    PubMed

    Ijichi, Nobuyuki; Kondo, Natsuko; Matsumoto, Rena; Shimada, Masakazu; Ishikawa, Hajime; Fukatsu, Takema

    2002-08-01

    The adzuki bean beetle, Callosobruchus chinensis, is infected with three distinct lineages of endosymbiotic bacteria belonging to the genus Wolbachia, which were designated wBruCon, wBruOri, and wBruAus. In an attempt to understand the mechanisms underlying the infection with these three organisms, the spatiotemporal infection dynamics of the three Wolbachia strains was investigated in detail by using a quantitative PCR technique. During the development of C. chinensis, the wBruCon, wBruOri, and wBruAus infection levels consistently increased but the growth patterns were different. The levels of infection plateaued at the pupal stage at approximately 3 x 10(8), 2 x 10(8), and 5 x 10(7) wsp copy equivalents per insect for wBruCon, wBruOri, and wBruAus, respectively. At the whole-insect level, the population densities of the three Wolbachia types did not show remarkable differences between adult males and females. At the tissue level, however, the total densities and relative levels of the three Wolbachia types varied significantly when different tissues and organs were compared and when the same tissues derived from males and females were compared. The histological data obtained by in situ hybridization and electron microscopy were concordant with the results of quantitative PCR analyses. Based on the histological data and the peculiar Wolbachia composition commonly found in nurse tissues and oocytes, we suggest that the Wolbachia strains are vertically transmitted to oocytes not directly, but by way of nurse tissue. On the basis of our results, we discuss interactions among the three coinfecting Wolbachia types, reproductive strategies of Wolbachia, and factors involved in the different cytoplasmic incompatibility phenotypes.

  16. The influence of passenger car front shape on pedestrian injury risk observed from German in-depth accident data.

    PubMed

    Li, Guibing; Lyons, Mathew; Wang, Bingyu; Yang, Jikuang; Otte, Dietmar; Simms, Ciaran

    2017-04-01

    odds increase by 74% for a 10% increase in NBLEH. These findings suggest that passenger car bumpers should support the lower leg with a low and flat lower bumper and even contact up to the femur area with a high upper bumper which extends above the knee to protect the pedestrian's leg. A low passenger car bonnet leading edge helps to reduce femur/pelvis injury risk. The passenger car front shape parameters are less influential than impact speed and pedestrian age for pedestrian injury risk.

  17. Characteristics, phenotype, and transmission of Wolbachia in the sweet potato whitefly, Bemisia tabaci (Hemiptera: Aleyrodidae), and its parasitoid Eretmocerus sp. nr. emiratus (Hymenoptera: Aphelinidae).

    PubMed

    Chiel, Elad; Kelly, Suzanne E; Harris, Alexandre M; Gebiola, Marco; Li, Xianchun; Zchori-Fein, Einat; Hunter, Martha S

    2014-04-01

    Wolbachia is a common intracellular bacterial endosymbiont of insects, causing a variety of effects including reproductive manipulations such as cytoplasmic incompatibility (CI). In this study, we characterized Wolbachia in the whitefly Bemisia tabaci and in the whitefly parasitoid Eretmocerus sp. nr. emiratus. We also tested for horizontal transmission of Wolbachia between and within trophic levels, and we determined the phenotype of Wolbachia in E. sp. nr. emiratus. Using multilocus sequence typing and phylogenetic analyses, we found that B. tabaci and E. sp. nr. emiratus each harbor a different and unique strain of Wolbachia. Both strains belong to the phylogenetic supergroup B. No evidence for horizontal transmission of Wolbachia between and within trophic levels was found in our study system. Finally, crossing results were consistent with a CI phenotype; when Wolbachia-infected E. sp. nr. emiratus males mate with uninfected females, wasp progeny survival dropped significantly, and the number of females was halved. This is the first description of CI caused by Wolbachia in the economically important genus Eretmocerus. Our study underscores the expectation that horizontal transmission events occur rarely in the dynamics of secondary symbionts such as Wolbachia, and highlights the importance of understanding the effects of symbionts on the biology of natural enemies.

  18. Wolbachia supergroups A and B in natural populations of medically important filth flies (diptera: muscidae, calliphoridae, and sarcophagidae) in Thailand.

    PubMed

    Mingchay, Pichanon; Sai-Ngam, Arkhom; Phumee, Atchara; Bhakdeenuan, Payu; Lorlertthum, Kittitouch; Thavara, Usavadee; Tawatsin, Apiwat; Choochote, Wej; Siriyasatien, Padet

    2014-03-01

    Filth flies, belonging to suborder Brachycera (Family; Muscidae, Calliphoridae and Sarcophagidae), are a major cause of nuisance and able to transmit pathogens to humans and animals. These insects are distributed worldwide and their populations are increasing especially in sub-tropical and tropical areas. One strategy for controlling insects employs Wolbachia, which is a group of maternally inherited intracellular bacteria, found in many insect species. The bacteria can cause reproductive abnormalities in their hosts, such as cytoplasmic incompatibility, feminization, parthenogenesis, and male lethality. In this study we determined Wolbachia endosymbionts in natural population of medically important flies (42 females and 9 males) from several geographic regions of Thailand. Wolbachia supergroups A or B were detected in 7 of female flies using PCR specific for wsp. Sequence analysis of wsp showed variations between and within the Wolbachia supergroup. Phylogenetics demonstrated that wsp is able to diverge between Wolbachia supergroups A and B. These data should be useful in future Wolbachia-based programs of fly control.

  19. The effect of Wolbachia-induced cytoplasmic incompatibility on host population size in natural and manipulated systems.

    PubMed Central

    Dobson, Stephen L; Fox, Charles W; Jiggins, Francis M

    2002-01-01

    Obligate, intracellular bacteria of the genus Wolbachia often behave as reproductive parasites by manipulating host reproduction to enhance their vertical transmission. One of these reproductive manipulations, cytoplasmic incompatibility, causes a reduction in egg-hatch rate in crosses between individuals with differing infections. Applied strategies based upon cytoplasmic incompatibility have been proposed for both the suppression and replacement of host populations. As Wolbachia infections occur within a broad range of invertebrates, these strategies are potentially applicable to a variety of medically and economically important insects. Here, we examine the interaction between Wolbachia infection frequency and host population size. We use a model to describe natural invasions of Wolbachia infections, artificial releases of infected hosts and releases of sterile males, as part of a traditional sterile insect technique programme. Model simulations demonstrate the importance of understanding the reproductive rate and intraspecific competition type of the targeted population, showing that releases of sterile or incompatible individuals may cause an undesired increase in the adult number. In addition, the model suggests a novel applied strategy that employs Wolbachia infections to suppress host populations. Releases of Wolbachia-infected hosts can be used to sustain artificially an unstable coexistence of multiple incompatible infections within a host population, allowing the host population size to be reduced, maintained at low levels, or eliminated. PMID:11886634

  20. The RNAi pathway plays a small part in Wolbachia-mediated blocking of dengue virus in mosquito cells

    PubMed Central

    Terradas, Gerard; Joubert, D. Albert; McGraw, Elizabeth A.

    2017-01-01

    Wolbachia pipientis is an insect endosymbiont known to limit the replication of viruses including dengue and Zika in their primary mosquito vector, Aedes aegypti. Wolbachia is being released into mosquito populations globally in a bid to control the diseases caused by these viruses. It is theorized that Wolbachia’s priming of the insect immune system may confer protection against subsequent viral infection. Other hypotheses posit a role for competition between Wolbachia and viruses for host cellular resources. Using an A. aegypti cell line infected with Wolbachia, we tested the effects of targeting siRNAs against the major innate immune pathways on dengue virus loads. We show that while Wolbachia infection induces genes in the Toll, JAK/STAT and RNAi pathways, only reduced expression of RNAi leads to a rebound of dengue virus loads in Wolbachia-infected cells. The magnitude of the effect explained less than 10% of the total DENV load, demonstrating that blocking must be dependent on other factors in addition to the expression of RNAi. The findings bode well for the long-term stability of blocking given that immunity gene expression would likely be highly plastic and susceptible to rapid evolution. PMID:28262718

  1. Wolbachia Affects Reproduction and Population Dynamics of the Coffee Berry Borer (Hypothenemus hampei): Implications for Biological Control

    PubMed Central

    Mariño, Yobana A.; Verle Rodrigues, José C.; Bayman, Paul

    2017-01-01

    Wolbachia are widely distributed endosymbiotic bacteria that influence the reproduction and fitness of their hosts. In recent years the manipulation of Wolbachia infection has been considered as a potential tool for biological control. The coffee berry borer (CBB), Hypothenemus hampei, is the most devastating coffee pest worldwide. Wolbachia infection in the CBB has been reported, but until now the role of Wolbachia in CBB reproduction and fitness has not been tested. To address this issue we reared the CBB in artificial diets with and without tetracycline (0.1% w/v) for ten generations. Tetracycline reduced significantly the relative proportion of Wolbachia in the CBB microbiota from 0.49% to 0.04%. This reduction affected CBB reproduction: females fed with tetracycline had significantly fewer progeny, lower fecundity, and fewer eggs per female. Tetracycline also reduced the population growth rate (λ), net reproductive rate (R0), and mean generation time (T) in CBB; the reduction in population growth was mostly due to variation in fertility, according to life time response experiments (LTREs) analysis. Our results suggest that Wolbachia contribute to the reproductive success of the CBB and their manipulation represents a possible approach to CBB biocontrol mediated by microbiome management. PMID:28085049

  2. Detection and characterization of Wolbachia infections in natural populations of aphids: is the hidden diversity fully unraveled?

    PubMed

    Augustinos, Antonis A; Santos-Garcia, Diego; Dionyssopoulou, Eva; Moreira, Marta; Papapanagiotou, Aristeidis; Scarvelakis, Marios; Doudoumis, Vangelis; Ramos, Silvia; Aguiar, Antonio F; Borges, Paulo A V; Khadem, Manhaz; Latorre, Amparo; Tsiamis, George; Bourtzis, Kostas

    2011-01-01

    Aphids are a serious threat to agriculture, despite being a rather small group of insects. The about 4,000 species worldwide engage in highly interesting and complex relationships with their microbial fauna. One of the key symbionts in arthropods is Wolbachia, an α-Proteobacterium implicated in many important biological processes and believed to be a potential tool for biological control. Aphids were thought not to harbour Wolbachia; however, current data suggest that its presence in aphids has been missed, probably due to the low titre of the infection and/or to the high divergence of the Wolbachia strains of aphids. The goal of the present study is to map the Wolbachia infection status of natural aphids populations, along with the characterization of the detected Wolbachia strains. Out of 425 samples from Spain, Portugal, Greece, Israel and Iran, 37 were found to be infected. Our results, based mainly on 16S rRNA gene sequencing, indicate the presence of two new Wolbachia supergroups prevailing in aphids, along with some strains belonging either to supergroup B or to supergroup A.

  3. Bidirectional cytoplasmic incompatibility induced by cross-order transfection of Wolbachia: implications for control of the host population.

    PubMed

    Zhong, Yong; Li, Zheng-Xi

    2014-10-01

    Wolbachia are widespread endosymbionts in arthropods and some nematodes. This genus of bacteria is known to manipulate host reproduction by inducing cytoplasmic incompatibility (CI). This important phenotype is implicated in the control of host populations since Wolbachia can suppress host populations through the induction of CI in a way similar to the sterile insect technique. Here, we identified a candidate CI-inducing Wolbachia strain from the parasitic wasp Scleroderma guani (wSguBJ) by sequencing and phylogenetic analysis. This Wolbachia strain was then isolated, purified, and artificially transfected into the new whitefly host Bemisia tabaci through nymphal microinjection. Infection frequency monitoring by molecular detection showed that 60-80 % of the offspring from transfected whitefly populations was infected with wSguBJ six generations after the transfer. Laboratory rearing experiments indicated that the artificial transfection caused no significant difference in the numbers of offspring between the transfected and naturally infected populations and had no significant detrimental effects on the development of transfected males, although the development of transfected females was delayed. Reciprocal crossings revealed that bidirectional CI was induced between the transfected and naturally infected whiteflies. These data indicated that the cross-order transfer of the heterologous Wolbachia strain by nymphal microinjection was successful. Mass release of the transfected males that could stably carry the heterologous Wolbachia without significant compromise of fecundity/development may provide an alternative approach to control of host populations.

  4. The Modulation of the Symbiont/Host Interaction between Wolbachia pipientis and Aedes fluviatilis Embryos by Glycogen Metabolism

    PubMed Central

    da Rocha Fernandes, Mariana; Martins, Renato; Pessoa Costa, Evenilton; Casagrande Pacidônio, Etiene; Araujo de Abreu, Leonardo; da Silva Vaz, Itabajara; Moreira, Luciano A.; da Fonseca, Rodrigo Nunes; Logullo, Carlos

    2014-01-01

    Wolbachia pipientis, a maternally transmitted bacterium that colonizes arthropods, may affect the general aspects of insect physiology, particularly reproduction. Wolbachia is a natural endosymbiont of Aedes fluviatilis, whose effects in embryogenesis and reproduction have not been addressed so far. In this context, we investigated the correlation between glucose metabolism and morphological alterations during A. fluviatilis embryo development in Wolbachia-positive (W+) and Wolbachia-negative (W−) mosquito strains. While both strains do not display significant morphological and larval hatching differences, larger differences were observed in hexokinase activity and glycogen contents during early and mid-stages of embryogenesis, respectively. To investigate if glycogen would be required for parasite-host interaction, we reduced Glycogen Synthase Kinase-3 (GSK-3) levels in adult females and their eggs by RNAi. GSK-3 knock-down leads to embryonic lethality, lower levels of glycogen and total protein and Wolbachia reduction. Therefore, our results suggest that the relationship between A. fluviatilis and Wolbachia may be modulated by glycogen metabolism. PMID:24926801

  5. The modulation of the symbiont/host interaction between Wolbachia pipientis and Aedes fluviatilis embryos by glycogen metabolism.

    PubMed

    da Rocha Fernandes, Mariana; Martins, Renato; Pessoa Costa, Evenilton; Pacidônio, Etiene Casagrande; Araujo de Abreu, Leonardo; da Silva Vaz, Itabajara; Moreira, Luciano A; da Fonseca, Rodrigo Nunes; Logullo, Carlos

    2014-01-01

    Wolbachia pipientis, a maternally transmitted bacterium that colonizes arthropods, may affect the general aspects of insect physiology, particularly reproduction. Wolbachia is a natural endosymbiont of Aedes fluviatilis, whose effects in embryogenesis and reproduction have not been addressed so far. In this context, we investigated the correlation between glucose metabolism and morphological alterations during A. fluviatilis embryo development in Wolbachia-positive (W+) and Wolbachia-negative (W-) mosquito strains. While both strains do not display significant morphological and larval hatching differences, larger differences were observed in hexokinase activity and glycogen contents during early and mid-stages of embryogenesis, respectively. To investigate if glycogen would be required for parasite-host interaction, we reduced Glycogen Synthase Kinase-3 (GSK-3) levels in adult females and their eggs by RNAi. GSK-3 knock-down leads to embryonic lethality, lower levels of glycogen and total protein and Wolbachia reduction. Therefore, our results suggest that the relationship between A. fluviatilis and Wolbachia may be modulated by glycogen metabolism.

  6. Survey of Endosymbionts in the Diaphorina citri Metagenome and Assembly of a Wolbachia wDi Draft Genome

    PubMed Central

    Saha, Surya; Hunter, Wayne B.; Reese, Justin; Morgan, J. Kent; Marutani-Hert, Mizuri; Huang, Hong; Lindeberg, Magdalen

    2012-01-01

    Diaphorina citri (Hemiptera: Psyllidae), the Asian citrus psyllid, is the insect vector of Ca. Liberibacter asiaticus, the causal agent of citrus greening disease. Sequencing of the D. citri metagenome has been initiated to gain better understanding of the biology of this organism and the potential roles of its bacterial endosymbionts. To corroborate candidate endosymbionts previously identified by rDNA amplification, raw reads from the D. citri metagenome sequence were mapped to reference genome sequences. Results of the read mapping provided the most support for Wolbachia and an enteric bacterium most similar to Salmonella. Wolbachia-derived reads were extracted using the complete genome sequences for four Wolbachia strains. Reads were assembled into a draft genome sequence, and the annotation assessed for the presence of features potentially involved in host interaction. Genome alignment with the complete sequences reveals membership of Wolbachia wDi in supergroup B, further supported by phylogenetic analysis of FtsZ. FtsZ and Wsp phylogenies additionally indicate that the Wolbachia strain in the Florida D. citri isolate falls into a sub-clade of supergroup B, distinct from Wolbachia present in Chinese D. citri isolates, supporting the hypothesis that the D. citri introduced into Florida did not originate from China. PMID:23166822

  7. Wolbachia Affects Reproduction and Population Dynamics of the Coffee Berry Borer (Hypothenemus hampei): Implications for Biological Control.

    PubMed

    Mariño, Yobana A; Verle Rodrigues, José C; Bayman, Paul

    2017-01-11

    Wolbachia are widely distributed endosymbiotic bacteria that influence the reproduction and fitness of their hosts. In recent years the manipulation of Wolbachia infection has been considered as a potential tool for biological control. The coffee berry borer (CBB), Hypothenemus hampei, is the most devastating coffee pest worldwide. Wolbachia infection in the CBB has been reported, but until now the role of Wolbachia in CBB reproduction and fitness has not been tested. To address this issue we reared the CBB in artificial diets with and without tetracycline (0.1% w/v) for ten generations. Tetracycline reduced significantly the relative proportion of Wolbachia in the CBB microbiota from 0.49% to 0.04%. This reduction affected CBB reproduction: females fed with tetracycline had significantly fewer progeny, lower fecundity, and fewer eggs per female. Tetracycline also reduced the population growth rate (λ), net reproductive rate (R₀), and mean generation time (T) in CBB; the reduction in population growth was mostly due to variation in fertility, according to life time response experiments (LTREs) analysis. Our results suggest that Wolbachia contribute to the reproductive success of the CBB and their manipulation represents a possible approach to CBB biocontrol mediated by microbiome management.

  8. Detection and Characterization of Wolbachia Infections in Natural Populations of Aphids: Is the Hidden Diversity Fully Unraveled?

    PubMed Central

    Augustinos, Antonis A.; Santos-Garcia, Diego; Dionyssopoulou, Eva; Moreira, Marta; Papapanagiotou, Aristeidis; Scarvelakis, Marios; Doudoumis, Vangelis; Ramos, Silvia; Aguiar, Antonio F.; Borges, Paulo A. V.; Khadem, Manhaz; Latorre, Amparo; Tsiamis, George; Bourtzis, Kostas

    2011-01-01

    Aphids are a serious threat to agriculture, despite being a rather small group of insects. The about 4,000 species worldwide engage in highly interesting and complex relationships with their microbial fauna. One of the key symbionts in arthropods is Wolbachia, an α-Proteobacterium implicated in many important biological processes and believed to be a potential tool for biological control. Aphids were thought not to harbour Wolbachia; however, current data suggest that its presence in aphids has been missed, probably due to the low titre of the infection and/or to the high divergence of the Wolbachia strains of aphids. The goal of the present study is to map the Wolbachia infection status of natural aphids populations, along with the characterization of the detected Wolbachia strains. Out of 425 samples from Spain, Portugal, Greece, Israel and Iran, 37 were found to be infected. Our results, based mainly on 16S rRNA gene sequencing, indicate the presence of two new Wolbachia supergroups prevailing in aphids, along with some strains belonging either to supergroup B or to supergroup A. PMID:22174869

  9. Identifying influential factors of business process performance using dependency analysis

    NASA Astrophysics Data System (ADS)

    Wetzstein, Branimir; Leitner, Philipp; Rosenberg, Florian; Dustdar, Schahram; Leymann, Frank

    2011-02-01

    We present a comprehensive framework for identifying influential factors of business process performance. In particular, our approach combines monitoring of process events and Quality of Service (QoS) measurements with dependency analysis to effectively identify influential factors. The framework uses data mining techniques to construct tree structures to represent dependencies of a key performance indicator (KPI) on process and QoS metrics. These dependency trees allow business analysts to determine how process KPIs depend on lower-level process metrics and QoS characteristics of the IT infrastructure. The structure of the dependencies enables a drill-down analysis of single factors of influence to gain a deeper knowledge why certain KPI targets are not met.

  10. Leveraging percolation theory to single out influential spreaders in networks

    NASA Astrophysics Data System (ADS)

    Radicchi, Filippo; Castellano, Claudio

    2016-06-01

    Among the consequences of the disordered interaction topology underlying many social, technological, and biological systems, a particularly important one is that some nodes, just because of their position in the network, may have a disproportionate effect on dynamical processes mediated by the complex interaction pattern. For example, the early adoption of a commercial product by an opinion leader in a social network may change its fate or just a few superspreaders may determine the virality of a meme in social media. Despite many recent efforts, the formulation of an accurate method to optimally identify influential nodes in complex network topologies remains an unsolved challenge. Here, we present the exact solution of the problem for the specific, but highly relevant, case of the susceptible-infected-removed (SIR) model for epidemic spreading at criticality. By exploiting the mapping between bond percolation and the static properties of the SIR model, we prove that the recently introduced nonbacktracking centrality is the optimal criterion for the identification of influential spreaders in locally tree-like networks at criticality. By means of simulations on synthetic networks and on a very extensive set of real-world networks, we show that the nonbacktracking centrality is a highly reliable metric to identify top influential spreaders also in generic graphs not embedded in space and for noncritical spreading.

  11. Leveraging percolation theory to single out influential spreaders in networks.

    PubMed

    Radicchi, Filippo; Castellano, Claudio

    2016-06-01

    Among the consequences of the disordered interaction topology underlying many social, technological, and biological systems, a particularly important one is that some nodes, just because of their position in the network, may have a disproportionate effect on dynamical processes mediated by the complex interaction pattern. For example, the early adoption of a commercial product by an opinion leader in a social network may change its fate or just a few superspreaders may determine the virality of a meme in social media. Despite many recent efforts, the formulation of an accurate method to optimally identify influential nodes in complex network topologies remains an unsolved challenge. Here, we present the exact solution of the problem for the specific, but highly relevant, case of the susceptible-infected-removed (SIR) model for epidemic spreading at criticality. By exploiting the mapping between bond percolation and the static properties of the SIR model, we prove that the recently introduced nonbacktracking centrality is the optimal criterion for the identification of influential spreaders in locally tree-like networks at criticality. By means of simulations on synthetic networks and on a very extensive set of real-world networks, we show that the nonbacktracking centrality is a highly reliable metric to identify top influential spreaders also in generic graphs not embedded in space and for noncritical spreading.

  12. Passenger comfort technology for system decision making

    NASA Technical Reports Server (NTRS)

    Conner, D. W.

    1980-01-01

    Decisions requiring passenger comfort technology were shown to depend on: the relationship between comfort and other factors (e.g., cost, urgency, alternate modes) in traveler acceptance of the systems, serving a selected market require technology to quantify effects of comfort versus offsetting factors in system acceptance. Public predict the maximum percentage of travelers who willingly accept the overall comfort of any trip ride. One or the other of these technology requirements apply to decisions on system design, operation and maintenance.

  13. Collisions with passenger cars and moose, Sweden.

    PubMed Central

    Björnstig, U; Eriksson, A; Thorson, J; Bylund, P O

    1986-01-01

    The number of collisions between motor vehicles and moose is increasing in many countries. Collisions with large, high animals such as moose cause typical rear- and downward deformation of the windshield pillars and front roof, most pronounced for small passenger cars; the injury risk increases with the deformation of the car. A strengthening of the windshield pillars and front roof and the use of antilacerative windshields would reduce the injury risk to car occupants. PMID:3953927

  14. Terrorist Watchlist Checks and Air Passenger Prescreening

    DTIC Science & Technology

    2009-12-30

    U.S. port of entry or at airport security checkpoints prior U.S. air carrier flights. For these purposes, CBP administers the Automated Targeting...Passenger Screening at Airport Security Checkpoints ................................. 14 9/11 Commission Recommendations and CAPPS II...individuals at either international ports of entries upon arrival at a U.S. port of entry or at airport security checkpoints prior U.S. air carrier

  15. Requirements for the crash protection of older vehicle passengers.

    PubMed

    Morris, Andrew; Welsh, Ruth; Hassan, Ahamedali

    2003-01-01

    This study compares injury outcomes in vehicle crashes involving different age groups of belted passengers. Two datasets were considered. Firstly, UK national data revealed that younger passengers are much more likely to be involved in crashes per million miles travelled compared to older passengers although older passengers are much more likely to be killed or seriously injured compared to younger passengers. Secondly, in-depth vehicle crash injury data were examined to determine some of the underlying reasons for the enhanced injury risk amongst older passengers. In crashes of approximately equal severity, the older passenger group were significantly more likely to be fatally injured in frontal crashes (p<0.001). However young passengers were as equally likely to be killed in struck-side crashes compared to the older group. The results also showed that older passengers sustained more serious injuries to the chest region in frontal crashes compared with the younger aged group (p<0.0001) and it is this body region that is particularly problematic. When the data were analysed further, it was found that a large proportion of passengers were female and that in the majority of cases, the seat belt was responsible for injury. Since by the year 2030, 1 in 4 persons will be aged over 65 in most OECD countries, the results suggest a need for intervention through vehicle design including in-vehicle crashworthiness systems that take into account reduced tolerance to impact with ageing.

  16. Hard Braking Events Among Novice Teenage Drivers By Passenger Characteristics

    PubMed Central

    Simons-Morton, Bruce G.; Ouimet, Marie Claude; Wang, Jing; Klauer, Sheila G.; Lee, Suzanne E.; Dingus, Thomas A.

    2010-01-01

    Summary In a naturalistic study of teenage drivers (N = 42) hard braking events of ≤−0.45 g were assessed over the first 6 months of licensure. A total of 1,721 hard braking events were recorded. The video footage of a sample (816) of these events was examined to evaluate validity and reasons for hard braking. Of these, 788 (96.6%) were estimated valid, of which 79.1% were due to driver misjudgment, 10.8% to risky driving behavior, 5.3% to legitimate evasive maneuvers, and 4.8% to distraction. Hard braking events per 10 trips and per 100 miles were compared across passenger characteristics. Hard braking rates per 10 trips among newly licensed teenagers during the first 6 months of licensure were significantly higher when driving with teen passengers and lower with adult passengers than driving alone; rates per 100 miles were lower with adult passengers than with no passengers. Further examination of the results indicates that rates of hard braking with teenage passengers were significantly higher compared with no passengers: 1) for male drivers; 2) during the first month of licensure. The data suggest that that novice teenage driving performance may not be as good or safe when driving alone or with teenage passengers than with adult passengers and provide support for the hypothesis that teenage passengers increase driving risks, particularly during the first month of licensure. PMID:21243109

  17. Hard Braking Events Among Novice Teenage Drivers By Passenger Characteristics.

    PubMed

    Simons-Morton, Bruce G; Ouimet, Marie Claude; Wang, Jing; Klauer, Sheila G; Lee, Suzanne E; Dingus, Thomas A

    2009-06-22

    In a naturalistic study of teenage drivers (N = 42) hard braking events of ≤-0.45 g were assessed over the first 6 months of licensure. A total of 1,721 hard braking events were recorded. The video footage of a sample (816) of these events was examined to evaluate validity and reasons for hard braking. Of these, 788 (96.6%) were estimated valid, of which 79.1% were due to driver misjudgment, 10.8% to risky driving behavior, 5.3% to legitimate evasive maneuvers, and 4.8% to distraction. Hard braking events per 10 trips and per 100 miles were compared across passenger characteristics. Hard braking rates per 10 trips among newly licensed teenagers during the first 6 months of licensure were significantly higher when driving with teen passengers and lower with adult passengers than driving alone; rates per 100 miles were lower with adult passengers than with no passengers. Further examination of the results indicates that rates of hard braking with teenage passengers were significantly higher compared with no passengers: 1) for male drivers; 2) during the first month of licensure. The data suggest that that novice teenage driving performance may not be as good or safe when driving alone or with teenage passengers than with adult passengers and provide support for the hypothesis that teenage passengers increase driving risks, particularly during the first month of licensure.

  18. Comparative Genomics of Two Closely Related Wolbachia with Different Reproductive Effects on Hosts

    PubMed Central

    Newton, Irene L.G.; Clark, Michael E.; Kent, Bethany N.; Bordenstein, Seth R.; Qu, Jiaxin; Richards, Stephen; Kelkar, Yogeshwar D.; Werren, John H.

    2016-01-01

    Wolbachia pipientis are obligate intracellular bacteria commonly found in many arthropods. They can induce various reproductive alterations in hosts, including cytoplasmic incompatibility, male-killing, feminization, and parthenogenetic development, and can provide host protection against some viruses and other pathogens. Wolbachia differ from many other primary endosymbionts in arthropods because they undergo frequent horizontal transmission between hosts and are well known for an abundance of mobile elements and relatively high recombination rates. Here, we compare the genomes of two closely related Wolbachia (with 0.57% genome-wide synonymous divergence) that differ in their reproductive effects on hosts. wVitA induces a sperm–egg incompatibility (also known as cytoplasmic incompatibility) in the parasitoid insect Nasonia vitripennis, whereas wUni causes parthenogenetic development in a different parasitoid, Muscidifurax uniraptor. Although these bacteria are closely related, the genomic comparison reveals rampant rearrangements, protein truncations (particularly in proteins predicted to be secreted), and elevated substitution rates. These changes occur predominantly in the wUni lineage, and may be due in part to adaptations by wUni to a new host environment, or its phenotypic shift to parthenogenesis induction. However, we conclude that the approximately 8-fold elevated synonymous substitution rate in wUni is due to a either an elevated mutation rate or a greater number of generations per year in wUni, which occurs in semitropical host species. We identify a set of genes whose loss or pseudogenization in the wUni lineage implicates them in the phenotypic shift from cytoplasmic incompatibility to parthenogenesis induction. Finally, comparison of these closely related strains allows us to determine the fine-scale mutation patterns in Wolbachia. Although Wolbachia are AT rich, mutation probabilities estimated from 4-fold degenerate sites are not AT biased, and

  19. Wolbachia, Sodalis and trypanosome co-infections in natural populations of Glossina austeni and Glossina pallidipes

    PubMed Central

    2013-01-01

    Background Tsetse flies harbor at least three bacterial symbionts: Wigglesworthia glossinidia, Wolbachia pipientis and Sodalis glossinidius. Wigglesworthia and Sodalis reside in the gut in close association with trypanosomes and may influence establishment and development of midgut parasite infections. Wolbachia has been shown to induce reproductive effects in infected tsetse. This study was conducted to determine the prevalence of these endosymbionts in natural populations of G. austeni and G. pallidipes and to assess the degree of concurrent infections with trypanosomes. Methods Fly samples analyzed originated from Kenyan coastal forests (trapped in 2009–2011) and South African G. austeni collected in 2008. The age structure was estimated by standard methods. G. austeni (n=298) and G. pallidipes (n= 302) were analyzed for infection with Wolbachia and Sodalis using PCR. Trypanosome infection was determined either by microscopic examination of dissected organs or by PCR amplification. Results Overall we observed that G. pallidipes females had a longer lifespan (70 d) than G. austeni (54 d) in natural populations. Wolbachia infections were present in all G. austeni flies analysed, while in contrast, this symbiont was absent from G. pallidipes. The density of Wolbachia infections in the Kenyan G. austeni population was higher than that observed in South African flies. The infection prevalence of Sodalis ranged from 3.7% in G. austeni to about 16% in G. pallidipes. Microscopic examination of midguts revealed an overall trypanosome infection prevalence of 6% (n = 235) and 5% (n = 552), while evaluation with ITS1 primers indicated a prevalence of about 13% (n = 296) and 10% (n = 302) in G. austeni and G. pallidipes, respectively. The majority of infections (46%) were with T. congolense. Co-infection with all three organisms was observed at 1% and 3.3% in G. austeni and G. pallidipes, respectively. Eleven out of the thirteen (85%) co-infected flies

  20. Comparative Genomics of Two Closely Related Wolbachia with Different Reproductive Effects on Hosts.

    PubMed

    Newton, Irene L G; Clark, Michael E; Kent, Bethany N; Bordenstein, Seth R; Qu, Jiaxin; Richards, Stephen; Kelkar, Yogeshwar D; Werren, John H

    2016-06-03

    Wolbachia pipientis are obligate intracellular bacteria commonly found in many arthropods. They can induce various reproductive alterations in hosts, including cytoplasmic incompatibility, male-killing, feminization, and parthenogenetic development, and can provide host protection against some viruses and other pathogens. Wolbachia differ from many other primary endosymbionts in arthropods because they undergo frequent horizontal transmission between hosts and are well known for an abundance of mobile elements and relatively high recombination rates. Here, we compare the genomes of two closely related Wolbachia (with 0.57% genome-wide synonymous divergence) that differ in their reproductive effects on hosts. wVitA induces a sperm-egg incompatibility (also known as cytoplasmic incompatibility) in the parasitoid insect Nasonia vitripennis, whereas wUni causes parthenogenetic development in a different parasitoid, Muscidifurax uniraptor Although these bacteria are closely related, the genomic comparison reveals rampant rearrangements, protein truncations (particularly in proteins predicted to be secreted), and elevated substitution rates. These changes occur predominantly in the wUni lineage, and may be due in part to adaptations by wUni to a new host environment, or its phenotypic shift to parthenogenesis induction. However, we conclude that the approximately 8-fold elevated synonymous substitution rate in wUni is due to a either an elevated mutation rate or a greater number of generations per year in wUni, which occurs in semitropical host species. We identify a set of genes whose loss or pseudogenization in the wUni lineage implicates them in the phenotypic shift from cytoplasmic incompatibility to parthenogenesis induction. Finally, comparison of these closely related strains allows us to determine the fine-scale mutation patterns in Wolbachia Although Wolbachia are AT rich, mutation probabilities estimated from 4-fold degenerate sites are not AT biased, and

  1. Aircraft sound quality for passenger comfort and enhanced product image

    NASA Astrophysics Data System (ADS)

    Wakefield, Gregory H.; Bultemeier, Eric J.; West, Erik; Angerer, James R.; Bhat, Waman V.

    2005-09-01

    Passenger cabin noise requirements for commercial airplanes are being expanded beyond the traditional focus on noise annoyance and speech intelligibility. There is increasing recognition that the passenger response to the cabin soundscape is much more complex; affecting perceptions of product quality, and impacting fatigue and comfort. Tailoring the soundscape for a preferred cabin environment requires the development of metrics that capture a range of passenger responses. In a preliminary exploration of potential metrics, a series of experiments were undertaken to investigate passenger preference for several classes of stationary and transient sounds within the passenger cabin. The design, implementation, and data analysis for these experiments is discussed, along with the application of results to enhance cabin comfort and to convey product quality. Key findings confirm what has been found in sound quality studies in other industries: sound level alone does not fully account for passenger preference.

  2. [Air transport biomechanical risk: reduced mobility passengers' handling].

    PubMed

    Draicchio, F; Campoli, G; Silvetti, A; Badellino, E; Forzano, F; Ranavolo, A; Iavicoli, S; Campagna, G; Raffaele, G; Gismondi, M

    2012-01-01

    As the airport traffic increases there is a continuous increase of passengers with different motor disabilities. Disabled passenger's assistance causes a biomechanical overload in airport workers. Some disabled passengers are classified by IATA as WCHC (wheel chair in cabin or Charlie). Our study, was performed in one of the most important Italian airport on Charlie passengers (about 10% of all assistances). We identified four critical points: 1) wheelchair and baggage moving (unstable load), 2) inclined ramps with worker's backwards steps and braked wheelchair to prevent passenger tipping or falling, 3) transfer from standard wheelchair to bicycle wheelchair, specifically designed for the aisle; 4.) transfer from bicycle wheelchair to aircraft seat. The last two points required sometimes to lift passengers over the armrest and positioning them on a window side seat, causing a serious increase of biomechanical load. For each critical point we have proposed technical and organizational measures to reduce airport worker's biomechanical risk.

  3. Wolbachia and cytoplasmic incompatibility in the California Culex pipiens mosquito species complex: parameter estimates and infection dynamics in natural populations.

    PubMed Central

    Rasgon, Jason L; Scott, Thomas W

    2003-01-01

    Before maternally inherited bacterial symbionts like Wolbachia, which cause cytoplasmic incompatibility (CI; reduced hatch rate) when infected males mate with uninfected females, can be used in a program to control vector-borne diseases it is essential to understand their dynamics of infection in natural arthropod vector populations. Our study had four goals: (1) quantify the number of Wolbachia strains circulating in the California Culex pipiens species complex, (2) investigate Wolbachia infection frequencies and distribution in natural California populations, (3) estimate the parameters that govern Wolbachia spread among Cx. pipiens under laboratory and field conditions, and (4) use these values to estimate equilibrium levels and compare predicted infection prevalence levels to those observed in nature. Strain-specific PCR, wsp gene sequencing, and crossing experiments indicated that a single Wolbachia strain infects Californian Cx. pipiens. Infection frequency was near or at fixation in all populations sampled for 2 years along a >1000-km north-south transect. The combined statewide infection frequency was 99.4%. Incompatible crosses were 100% sterile under laboratory and field conditions. Sterility decreased negligibly with male age in the laboratory. Infection had no significant effect on female fecundity under laboratory or field conditions. Vertical transmission was >99% in the laboratory and approximately 98.6% in the field. Using field data, models predicted that Wolbachia will spread to fixation if infection exceeds an unstable equilibrium point above 1.4%. Our estimates accurately predicted infection frequencies in natural populations. If certain technical hurdles can be overcome, our data indicate that Wolbachia can invade vector populations as part of an applied transgenic strategy for vector-borne disease reduction. PMID:14704183

  4. Decapitation Improves Detection of Wolbachia pipientis (Rickettsiales: Anaplasmataceae) in Culex pipiens (Diptera: Culicidae) Mosquitoes by the Polymerase Chain Reaction

    PubMed Central

    BECKMANN, J. F.; FALLON, A. M.

    2013-01-01

    Polymerase chain reaction (PCR) is often used to detect microorganisms, pathogens, or both, including the reproductive parasite Wolbachia pipientis (Rickettsiales: Anaplasmataceae), in mosquitoes. Natural populations of Culex pipiens L. (Diptera: Culicidae) mosquitoes are infected with one or more strains of W. pipientis, and crosses between mosquitoes harboring different Wolbachia strains provide one of the best-known examples of cytoplasmic incompatibililty (CI). When we used PCR to monitor Wolbachia in the Buckeye strain of Culex pipiens, and in a Wolbachia-cured sister colony obtained by tetracycline treatment, we noted false negative PCR reactions with DNA samples from infected mosquitoes; these results were inconsistent with direct microscopic observation of Wolbachia-like particles in gonads dissected from mosquitoes in the same population. Assays with diluted template often improved detection of positive samples, suggesting that DNA prepared from whole mosquitoes contained an inhibitor of the PCR reaction. We reconciled discrepancies between PCR and microscopy by systematic measurement of the PCR reaction in the presence of an internal standard. Mosquito decapitation before DNA extraction restored the reliability of the PCR reaction, allowing accurate determination of Wolbachia infection status in infected and tetracycline-cured mosquito populations, consistent with microscopic examination. Using PCR primers based on the Tr1 gene, we confirmed that the Wolbachia infection in the Buckeye strain of Culex pipiens belongs to the genotype designated wPip1. Finally, to explore more widely the distribution of PCR inhibitors, we demonstrated that DNA isolated from the cricket, Acheta domesticus (L.); the beetle, Tenebrio molitor L.; the honey bee, Apis mellifera L.; and the mosquito, Anopheles punctipennis Say also contained PCR inhibitors. These results underscore the importance of measuring the presence of inhibitors in PCR templates by using a known positive

  5. Wolbachia Blocks Viral Genome Replication Early in Infection without a Transcriptional Response by the Endosymbiont or Host Small RNA Pathways

    PubMed Central

    McFarlane, Melanie; Juneja, Punita; Sarkies, Peter; Lulla, Aleksei; Schnettler, Esther; Varjak, Margus; Merits, Andres; Miska, Eric A.; Jiggins, Francis M.; Kohl, Alain

    2016-01-01

    The intracellular endosymbiotic bacterium Wolbachia can protect insects against viral infection, and is being introduced into mosquito populations in the wild to block the transmission of arboviruses that infect humans and are a major public health concern. To investigate the mechanisms underlying this antiviral protection, we have developed a new model system combining Wolbachia-infected Drosophila melanogaster cell culture with the model mosquito-borne Semliki Forest virus (SFV; Togaviridae, Alphavirus). Wolbachia provides strong antiviral protection rapidly after infection, suggesting that an early stage post-infection is being blocked. Wolbachia does appear to have major effects on events distinct from entry, assembly or exit as it inhibits the replication of an SFV replicon transfected into the cells. Furthermore, it causes a far greater reduction in the expression of proteins from the 3´ open reading frame than the 5´ non-structural protein open reading frame, indicating that it is blocking the replication of viral RNA. Further to this separation of the replicase proteins and viral RNA in transreplication assays shows that uncoupling of viral RNA and replicase proteins does not overcome Wolbachia’s antiviral activity. This further suggests that replicative processes are disrupted, such as translation or replication, by Wolbachia infection. This may occur by Wolbachia mounting an active antiviral response, but the virus did not cause any transcriptional response by the bacterium, suggesting that this is not the case. Host microRNAs (miRNAs) have been implicated in protection, but again we found that host cell miRNA expression was unaffected by the bacterium and neither do our findings suggest any involvement of the antiviral siRNA pathway. We conclude that Wolbachia may directly interfere with early events in virus replication such as translation of incoming viral RNA or RNA transcription, and this likely involves an intrinsic (as opposed to an induced

  6. Two Strains of Male-Killing Wolbachia in a Ladybird, Coccinella undecimpunctata, from a Hot Climate

    PubMed Central

    Elnagdy, Sherif; Messing, Susan

    2013-01-01

    Ladybirds are a hot-spot for the invasion of male-killing bacteria. These maternally inherited endosymbionts cause the death of male host embryos, to the benefit of female sibling hosts and the bacteria that they contain. Previous studies have shown that high temperatures can eradicate male-killers from ladybirds, leaving the host free from infection. Here we report the discovery of two maternally inherited sex ratio distorters in populations of a coccinellid, Coccinella undecimpunctata, from a hot lowland region of the Middle East. DNA sequence analysis indicates that the male killing is the result of infection by Wolbachia, that the trait is tetracycline sensitive, and that two distinct strains of Wolbachia co-occur within one beetle population. We discuss the implications of these findings for theories of male-killing and suggest avenues for future field-work on this system. PMID:23349831

  7. Two strains of male-killing Wolbachia in a ladybird, Coccinella undecimpunctata, from a hot climate.

    PubMed

    Elnagdy, Sherif; Messing, Susan; Majerus, Michael E N

    2013-01-01

    Ladybirds are a hot-spot for the invasion of male-killing bacteria. These maternally inherited endosymbionts cause the death of male host embryos, to the benefit of female sibling hosts and the bacteria that they contain. Previous studies have shown that high temperatures can eradicate male-killers from ladybirds, leaving the host free from infection. Here we report the discovery of two maternally inherited sex ratio distorters in populations of a coccinellid, Coccinella undecimpunctata, from a hot lowland region of the Middle East. DNA sequence analysis indicates that the male killing is the result of infection by Wolbachia, that the trait is tetracycline sensitive, and that two distinct strains of Wolbachia co-occur within one beetle population. We discuss the implications of these findings for theories of male-killing and suggest avenues for future field-work on this system.

  8. Dirofilaria immitis and Wolbachia pipientis: a thorough investigation of the symbiosis responsible for canine heartworm disease.

    PubMed

    McHaffie, Jake

    2012-02-01

    Canine heartworm disease wreaks havoc inside canines all throughout the modern world, including the USA. Any region where mosquitoes thrive will provide efficient dog-to-dog transportation for the microfilaria of the infectious nematode Dirofilaria immitis. Veterinary scientists have recently discovered both phylogenetic and biochemical evidence for the obligate symbiosis of D. immitis and the bacteria Wolbachia pipientis. As a result, veterinarians have initiated testing of antibiotic therapies either instead of, or together with, the currently utilized antiparasitic treatments for canine heartworm. The toxicity of melarsomine adulticidal therapies has prompted an abundance of new research involving doxycycline and other antibiotics, which will be addressed in this review. As our knowledge of the Wolbachia endosymbiont expands, so will our abilities to minimize toxicity and maximize efficiency of heartworm treatments.

  9. Vehicle hydraulic system that provides heat for passenger compartment

    DOEpatents

    Bartley, Bradley E.; Blass, James R.; Gibson, Dennis H.

    2001-01-01

    A vehicle includes a vehicle housing which defines a passenger compartment. Attached to the vehicle housing is a hydraulic system, that includes a hydraulic fluid which flows through at least one passageway within the hydraulic system. Also attached to the vehicle housing is a passenger compartment heating system. The passenger compartment heating system includes a heat exchanger, wherein a portion of the heat exchanger is a segment of the at least one passageway of the hydraulic system.

  10. Passenger leukocytes and microchimerism: what role in tolerance induction?

    PubMed

    Wood, Kathryn J

    2003-05-15

    The role of passenger leukocytes in determining the outcome after transplantation is complex. In some settings, donor-derived passenger leukocytes can initiate graft rejection, whereas in others they contribute to graft acceptance. Both donor and recipient factors contribute to this potential dual role. Understanding the interaction between passenger leukocytes and the recipient's immune system, particularly after liver transplantation, may provide important clues for developing novel strategies for inducing specific unresponsiveness to donor alloantigens.

  11. A Re-Examination of Wolbachia-Induced Cytoplasmic Incompatibility in California Drosophila simulans

    PubMed Central

    Carrington, Lauren B.; Lipkowitz, Jeremy R.; Hoffmann, Ary A.; Turelli, Michael

    2011-01-01

    Background In California Drosophila simulans, the maternally inherited Riverside strain Wolbachia infection (wRi) provides a paradigm for rapid spread of Wolbachia in nature and rapid evolutionary change. wRi induces cytoplasmic incompatibility (CI), where crosses between infected males and uninfected females produce reduced egg-hatch. The three parameters governing wRi infection-frequency dynamics quantify: the fidelity of maternal transmission, the level of cytoplasmic incompatibility, and the relative fecundity of infected females. We last estimated these parameters in nature in 1993. Here we provide new estimates, under both field and laboratory conditions. Five years ago, we found that wRi had apparently evolved over 15 years to enhance the fecundity of infected females; here we examine whether CI intensity has also evolved. Methodology/Principal Findings New estimates using wild-caught flies indicate that the three key parameters have remained relatively stable since the early 1990s. As predicted by our three-parameter model using field-estimated parameter values, population infection frequencies remain about 93%. Despite this relative stability, laboratory data based on reciprocal crosses and introgression suggest that wRi may have evolved to produce less intense CI (i.e., higher egg hatch from incompatible crosses). In contrast, we find no evidence that D. simulans has evolved to lower the susceptibility of uninfected females to CI. Conclusions/Significance Evolution of wRi that reduces CI is consistent with counterintuitive theoretical predictions that within-population selection on CI-causing Wolbachia does not act to increase CI. Within taxa, CI is likely to evolve mainly via pleiotropic effects associated with the primary targets of selection on Wolbachia, i.e., host fecundity and transmission fidelity. Despite continuous, strong selection, D. simulans has not evolved appreciably to suppress CI. Our data demonstrate a lack of standing genetic variation

  12. A New Model and Method for Understanding Wolbachia-Induced Cytoplasmic Incompatibility

    PubMed Central

    Bossan, Benjamin; Koehncke, Arnulf; Hammerstein, Peter

    2011-01-01

    Wolbachia are intracellular bacteria transmitted almost exclusively vertically through eggs. In response to this mode of transmission, Wolbachia strategically manipulate their insect hosts' reproduction. In the most common manipulation type, cytoplasmic incompatibility, infected males can only mate with infected females, but infected females can mate with all males. The mechanism of cytoplasmic incompatibility is unknown; theoretical and empirical findings need to converge to broaden our understanding of this phenomenon. For this purpose, two prominent models have been proposed: the mistiming-model and the lock-key-model. The former states that Wolbachia manipulate sperm of infected males to induce a fatal delay of the male pronucleus during the first embryonic division, but that the bacteria can compensate the delay by slowing down mitosis in fertilized eggs. The latter states that Wolbachia deposit damaging “locks” on sperm DNA of infected males, but can also provide matching “keys” in infected eggs to undo the damage. The lock-key-model, however, needs to assume a large number of locks and keys to explain all existing incompatibility patterns. The mistiming-model requires fewer assumptions but has been contradicted by empirical results. We therefore expand the mistiming-model by one quantitative dimension to create the new, so-called goalkeeper-model. Using a method based on formal logic, we show that both lock-key- and goalkeeper-model are consistent with existing data. Compared to the lock-key-model, however, the goalkeeper-model assumes only two factors and provides an idea of the evolutionary emergence of cytoplasmic incompatibility. Available cytological evidence suggests that the hypothesized second factor of the goalkeeper-model may indeed exist. Finally, we suggest empirical tests that would allow to distinguish between the models. Generalizing our results might prove interesting for the study of the mechanism and evolution of other host

  13. Wolbachia-Induced Unidirectional Cytoplasmic Incompatibility and Speciation: Mainland-Island Model

    PubMed Central

    Telschow, Arndt; Flor, Matthias; Kobayashi, Yutaka; Hammerstein, Peter; Werren, John H.

    2007-01-01

    Bacteria of the genus Wolbachia are among the most common endosymbionts in the world. In many insect species these bacteria induce a sperm-egg incompatibility between the gametes of infected males and uninfected females, commonly called unidirectional cytoplasmic incompatibility (CI). It is generally believed that unidirectional CI cannot promote speciation in hosts because infection differences between populations will be unstable and subsequent gene flow will eliminate genetic differences between diverging populations. In the present study we investigate this question theoretically in a mainland-island model with migration from mainland to island. Our analysis shows that (a) the infection polymorphism is stable below a critical migration rate, (b) an (initially) uninfected “island” can better maintain divergence at a selected locus (e.g. can adapt locally) in the presence of CI, and (c) unidirectional CI selects for premating isolation in (initially) uninfected island populations if they receive migration from a Wolbachia-infected mainland. Interestingly, premating isolation is most likely to evolve if levels of incompatibility are intermediate and if either the infection causes fecundity reductions or Wolbachia transmission is incomplete. This is because under these circumstances an infection pattern with an infected mainland and a mostly uninfected island can persist in the face of comparably high migration. We present analytical results for all three findings: (a) a lower estimation of the critical migration rate in the presence of local adaptation, (b) an analytical approximation for the gene flow reduction caused by unidirectional CI, and (c) a heuristic formula describing the invasion success of mutants at a mate preference locus. These findings generally suggest that Wolbachia-induced unidirectional CI can be a factor in divergence and speciation of hosts. PMID:17684548

  14. The Effect of Temperature on Wolbachia-Mediated Dengue Virus Blocking in Aedes aegypti.

    PubMed

    Ye, Yixin H; Carrasco, Alison M; Dong, Yi; Sgrò, Carla M; McGraw, Elizabeth A

    2016-04-01

    Dengue fever, caused by dengue virus (DENV), is endemic in more than 100 countries. The lack of effective treatment of patients and the suboptimal efficacies of the tetravalent vaccine in trials highlight the urgent need to develop alternative strategies to lessen the burden of dengue fever.Wolbachia pipientis, an obligate intracellular bacterium, is being developed as a biocontrol strategy against dengue because it limits the replication of the DENV in the mosquito vector,Aedes aegypti However, several recent studies have demonstrated the sensitivity of pathogens, vectors, and their symbionts to temperature. To understand how the tripartite interactions between the mosquito, DENV, and Wolbachia may change under different temperature regimes, we assessed the vector competence and transmission potential of DENV-infected mosquitoes reared at a common laboratory setting of a constant 25°C and at two diurnal temperature settings with mean of 25°C and 28°C and a fluctuating range of 8°C (±4°C). Temperature significantly affected DENV infection rate in the mosquitoes. Furthermore, temperature significantly influenced the proportion of mosquitoes that achieved transmission potential as measured by the presence of virus in the saliva. Regardless of the temperature regimes,Wolbachia significantly and efficiently reduced the proportion of mosquitoes achieving infection and transmission potential across all the temperature regimes studied. This work reinforces the robustness of the Wolbachia biocontrol strategy to field conditions in Cairns, Australia, and suggests that similar studies are required for local mosquito genotypes and field relevant temperatures for emerging field release sites globally.

  15. 16. UPPER PASSENGER LOADING ZONE, NOTE LOADING ZONE BELOW WITH ...

    Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

    16. UPPER PASSENGER LOADING ZONE, NOTE LOADING ZONE BELOW WITH TURN STILES - Jefferson National Expansion Memorial Arch, Mississippi River between Washington & Poplar Streets, Saint Louis, Independent City, MO

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

    PubMed Central

    White, Jennifer A.; Kelly, Suzanne E.; Perlman, Steve J.; Hunter, Martha S.

    2014-01-01

    Many bacterial endosymbionts of insects are capable of manipulating their host’s 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. But 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. PMID:19223921

  17. Wolbachia are present in southern african scorpions and cluster with supergroup F.

    PubMed

    Baldo, Laura; Prendini, Lorenzo; Corthals, Angelique; Werren, John H

    2007-11-01

    The presence and distribution of the intracellular bacteria Wolbachia in the arthropod subphylum Chelicerata (including class Arachnida) has not been extensively explored. Here we report the discovery of Wolbachia in scorpions. Five strains found in host species of the genus Opistophthalmus (Southern African burrowing scorpions) have been characterized by Multilocus Sequence Typing and by Wolbachia Surface Protein. Phylogenetic analyses indicate clustering in the supergroup F and a high genetic relatedness among all scorpion strains as a result of a potential transmission within the host genus. The F-group is an uncommon lineage compared to the A and B supergroups, although it is present in a broad range of hosts (including insects, filarial nematodes, and now arachnids) and across a large geographical area (e.g., North America, Africa, Europe, and Australia). It also shows no evidence of recombination and has a significantly higher genetic diversity than supergroup A and B. Overall, this pattern suggests an older radiation of F-strains with respect to A and B-strains, followed by limited horizontal transmission across host genera and reduced genetic flux among strains. A more extensive sampling of supergroup F-strains is required to confirm this scenario.

  18. Serum antibody responses to Wolbachia surface protein in patients with human lymphatic filariasis.

    PubMed

    Shiny, Chandanapurath; Krushna, Nagampalli S A; Archana, Bairavasundaram; Farzana, Begum; Narayanan, Rangarajan B

    2009-12-01

    Wolbachia surface protein (WSP), which is the most abundantly expressed protein of Wolbachia from the human filarial parasite Brugia malayi, was chosen for the present study. B-cell epitope prediction of the WSP protein sequence indicates a high antigenicity, surface probability and hydrophilicity by DNA STAR software analysis. ProPred analysis suggests the presence of HLA class II binding regions in the WSP protein that contribute to T-cell responses and isotype reactivity. In order to validate these findings, the gene coding for endosymbiont WSP was PCR-amplified from the genomic DNA of the human filarial parasite Brugia malayi and cloned in T-7 expression vector pRSET-A. Western blot and ELISA at the total IgG level with recombiant WSP indicated a significantly elevated reactivity in CP compared to MF, EN and NEN individuals. Isotype ELISA also suggested an elevated reactivity in CP patients at the IgG1 level. In contrast, WSP-specific IgG4 levels were found to be elevated in MF patients compared to CP and EN. Besides this, WSP-specific IgE levels indicated an elevated reactivity in CP and MF patients compared to normals. Observations from ELISA supported the in silico predictions that indicate the presence of B- and T-cell epitopes. Hence, a combinatorial approach of in silico predictions and wet-lab studies provides interesting insights into the role of Wolbachia proteins in filarial pathogenesis.

  19. [Detection and analysis of symbiotic bacteria, Arsenophonus and Wolbachia, in striped stem borer, Chilo suppressalis (Lepidoptera: Crambidae)].

    PubMed

    Chai, Huan-Na; Wu, Hai-Yan; Du, Yu-Zhou

    2013-12-01

    In this study, 23S rDNA from Arsenophonus and wsp gene from Wolbachia were amplified by PCR method using specific primers to study their symbiosis in Chilo suppressalis from 20 locations of China. The results indicated that Arsenophonus and Wolbachia infection rates differed widely in C. suppressalis populations. The infection rates of Arsenophonus ranged from 5.0% to 50.0% in five geographical populations from Harbin, Huishui, Jilin, Nanyang, and Yangzhou. Wolbachia ranged from 25.0% to 40.0% in three geographical populations from Hanzhong, Nanning, and Yangzhou. Their symbiosis was not observed in C. suppressalis of other locations. The sequences of 23S rDNA gene which named csArs were exactly the same in the five populations. However, the sequences of wsp from the three strains of Wolbachia showed wChisup1 belonged to supergroup A, wChisup5 and wChisup6 belonged to supergroup B. The results showed the strains of Arsenophonus from C. suppressalis of the five different locations were identical, whereas the strains of Wolbachia were phylogenetically diverse. The phylogenetic analysis indicated that 23S rDNA and wsp sequences found in C. suppressalis were exactly the same with or very closely related to relevant sequences in other species.

  20. Impact of population age structure on Wolbachia transgene driver efficacy: ecologically complex factors and release of genetically modified mosquitoes.

    PubMed

    Rasgon, Jason L; Scott, Thomas W

    2004-07-01

    Wolbachia symbionts hold theoretical promise as a way to drive transgenes into insect vector populations for disease prevention. For simplicity, current models of Wolbachia dynamics and spread ignore ecologically complex factors such as the age structure of vector populations and overlapping vector generations. We developed a model including these factors to assess their impact on the process of Wolbachia spread into populations of three mosquito species (Anopheles gambiae, Aedes aegypti and Culex pipiens). Depending on the mosquito species, Wolbachia parameters, released mosquito life stage and initial age structure of the target population, the number of Wolbachia-infected mosquitoes that we predict would need to be released ranged from less than the threshold calculated by the simple model to a 10-30-fold increase. Transgenic releases into age-structured populations, which is an expectation for wild mosquitoes, will be difficult and depending on the circumstances may not be economically or logistically feasible due to the large number of infected mosquitoes that must be released. Our results support the perspective that understanding ecological factors is critical for designing transgenic vector-borne disease control strategies.

  1. Screening of Wolbachia endosymbiont infection in Aedes aegypti mosquitoes using Attenuated Total Reflection mid-infrared spectroscopy.

    PubMed

    Khoshmanesh, Aazam; Christensen, Dale; Perez-Guaita, David; Iturbe-Ormaetxe, Iñaki; O'Neill, Scott L; McNaughton, Don; Wood, Bayden R

    2017-03-23

    Dengue fever is the most common mosquito transmitted viral infection afflicting humans, estimated to generate around 390 million infections each year in over 100 countries. The introduction of the endosymbiotic bacterium Wolbachia into Aedes aegypti mosquitoes has the potential to greatly reduce the public health burden of the disease. This approach requires extensive PCR (Polymerase Chain Reaction) testing of the Wolbachia-infection status of mosquitoes in areas where Wolbachia-A. aegypti are released. Here we report the first example of small organism mid-infrared spectroscopy where we have applied Attenuated Total Reflection Fourier Transform Infrared (ATR-FTIR) spectroscopy and multivariate modelling methods to determine sex, age and the presence of Wolbachia (wMel strain) in laboratory mosquitoes and sex and age in field mosquitoes. The prediction errors using Partial Least Squares Discriminant Analysis (PLS-DA) discrimination models for laboratory studies on independent test sets ranged from 0 to 3% for age & sex grading, and 3 to 5% for Wolbachia infection diagnosis using dry mosquito abdomens while field study results using an Artificial neural network yielded a 10 % error. The application of FTIR analysis is inexpensive, easy to use, portable, and shows significant potential to replace the reliance on more expensive and laborious PCR assays.

  2. Removing the needle from the haystack: Enrichment of Wolbachia endosymbiont transcripts from host nematode RNA by Cappable-seq™.

    PubMed

    Luck, Ashley N; Slatko, Barton E; Foster, Jeremy M

    2017-01-01

    Efficient transcriptomic sequencing of microbial mRNA derived from host-microbe associations is often compromised by the much lower relative abundance of microbial RNA in the mixed total RNA sample. One solution to this problem is to perform extensive sequencing until an acceptable level of transcriptome coverage is obtained. More cost-effective methods include use of prokaryotic and/or eukaryotic rRNA depletion strategies, sometimes in conjunction with depletion of polyadenylated eukaryotic mRNA. Here, we report use of Cappable-seq™ to specifically enrich, in a single step, Wolbachia endobacterial mRNA transcripts from total RNA prepared from the parasitic filarial nematode, Brugia malayi. The obligate Wolbachia endosymbiont is a proven drug target for many human filarial infections, yet the precise nature of its symbiosis with the nematode host is poorly understood. Insightful analysis of the expression levels of Wolbachia genes predicted to underpin the mutualistic association and of known drug target genes at different life cycle stages or in response to drug treatments is typically challenged by low transcriptomic coverage. Cappable-seq resulted in up to ~ 5-fold increase in the number of reads mapping to Wolbachia. On average, coverage of Wolbachia transcripts from B. malayi microfilariae was enriched ~40-fold by Cappable-seq. Additionally, this method has an additional benefit of selectively removing abundant prokaryotic ribosomal RNAs.The deeper microbial transcriptome sequencing afforded by Cappable-seq facilitates more detailed characterization of gene expression levels of pathogens and symbionts present in animal tissues.

  3. Wolbachia infections mimic cryptic speciation in two parasitic butterfly species, Phengaris teleius and P. nausithous (Lepidoptera: Lycaenidae).

    PubMed

    Ritter, Sylvia; Michalski, Stefan G; Settele, Josef; Wiemers, Martin; Fric, Zdenek F; Sielezniew, Marcin; Šašić, Martina; Rozier, Yves; Durka, Walter

    2013-01-01

    Deep mitochondrial divergence within species may result from cryptic speciation, from phylogeographic isolation or from endosymbiotic bacteria like Wolbachia that manipulate host reproduction. Phengaris butterflies are social parasites that spend most of their life in close relationship with ants. Previously, cryptic speciation has been hypothesised for two Phengaris species based on divergent mtDNA sequences. Since Phengaris species are highly endangered, the existence of cryptic species would have drastic consequences for conservation and management. We tested for cryptic speciation and alternative scenarios in P. teleius and P. nausithous based on a comprehensive sample across their Palaearctic ranges using COI gene sequences, nuclear microsatellites and tests for Wolbachia. In both species a deep mitochondrial split occurring 0.65-1.97 myrs ago was observed that did not correspond with microsatellite data but was concordant with Wolbachia infection. Haplotypes previously attributed to cryptic species were part of the Wolbachia-infected clades. In both species remaining phylogeographic structure was largely consistent between mitochondrial and nuclear genomes. In P. teleius several mitochondrial and nuclear groups were observed in East Asia while a single haplogroup and nuclear cluster prevailed across continental Eurasia. Neutrality tests suggested rapid demographic expansion into that area. In contrast, P. nausithous had several mitochondrial and nuclear groups in Europe, suggesting a complex phylogeographic history in the western part of the species range. We conclude that deep intraspecific divergences found in DNA barcode studies do not necessarily need to represent cryptic speciation but instead can be due to both infection by Wolbachia and phylogeographic structure.

  4. 75 FR 7599 - Notice of Public Hearing; Passenger Vessel Financial Responsibility

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-02-22

    ... Notice of Public Hearing; Passenger Vessel Financial Responsibility AGENCY: Federal Maritime Commission... Passenger Vessel Financial Responsibility Program. DATES: The due date for submitting a request to... make presentations concerning the Commission's passenger vessel financial responsibility program....

  5. Identify influential spreaders in complex networks, the role of neighborhood

    NASA Astrophysics Data System (ADS)

    Liu, Ying; Tang, Ming; Zhou, Tao; Do, Younghae

    2016-06-01

    Identifying the most influential spreaders is an important issue in controlling the spreading processes in complex networks. Centrality measures are used to rank node influence in a spreading dynamics. Here we propose a node influence measure based on the centrality of a node and its neighbors' centrality, which we call the neighborhood centrality. By simulating the spreading processes in six real-world networks, we find that the neighborhood centrality greatly outperforms the basic centrality of a node such as the degree and coreness in ranking node influence and identifying the most influential spreaders. Interestingly, we discover a saturation effect in considering the neighborhood of a node, which is not the case of the larger the better. Specifically speaking, considering the 2-step neighborhood of nodes is a good choice that balances the cost and performance. If further step of neighborhood is taken into consideration, there is no obvious improvement and even decrease in the ranking performance. The saturation effect may be informative for studies that make use of the local structure of a node to determine its importance in the network.

  6. Discovering Influential Nodes for SIS Models in Social Networks

    NASA Astrophysics Data System (ADS)

    Saito, Kazumi; Kimura, Masahiro; Motoda, Hiroshi

    We address the problem of efficiently discovering the influential nodes in a social network under the susceptible/infected/susceptible (SIS) model, a diffusion model where nodes are allowed to be activated multiple times. The computational complexity drastically increases because of this multiple activation property. We solve this problem by constructing a layered graph from the original social network with each layer added on top as the time proceeds, and applying the bond percolation with pruning and burnout strategies. We experimentally demonstrate that the proposed method gives much better solutions than the conventional methods that are solely based on the notion of centrality for social network analysis using two large-scale real-world networks (a blog network and a wikipedia network). We further show that the computational complexity of the proposed method is much smaller than the conventional naive probabilistic simulation method by a theoretical analysis and confirm this by experimentation. The properties of the influential nodes discovered are substantially different from those identified by the centrality-based heuristic methods.

  7. Identifying influential directors in the United States corporate governance network.

    PubMed

    Huang, Xuqing; Vodenska, Irena; Wang, Fengzhong; Havlin, Shlomo; Stanley, H Eugene

    2011-10-01

    The influence of directors has been one of the most engaging topics recently, but surprisingly little research has been done to quantitatively evaluate the influence and power of directors. We analyze the structure of the US corporate governance network for the 11-year period 1996-2006 based on director data from the Investor Responsibility Research Center director database, and we develop a centrality measure named the influence factor to estimate the influence of directors quantitatively. The US corporate governance network is a network of directors with nodes representing directors and links between two directors representing their service on common company boards. We assume that information flows in the network through information-sharing processes among linked directors. The influence factor assigned to a director is based on the level of information that a director obtains from the entire network. We find that, contrary to commonly accepted belief that directors of large companies, measured by market capitalization, are the most powerful, in some instances, the directors who are influential do not necessarily serve on boards of large companies. By applying our influence factor method to identify the influential people contained in the lists created by popular magazines such as Fortune, Networking World, and Treasury and Risk Management, we find that the influence factor method is consistently either the best or one of the two best methods in identifying powerful people compared to other general centrality measures that are used to denote the significance of a node in complex network theory.

  8. Identifying influential directors in the United States corporate governance network

    NASA Astrophysics Data System (ADS)

    Huang, Xuqing; Vodenska, Irena; Wang, Fengzhong; Havlin, Shlomo; Stanley, H. Eugene

    2011-10-01

    The influence of directors has been one of the most engaging topics recently, but surprisingly little research has been done to quantitatively evaluate the influence and power of directors. We analyze the structure of the US corporate governance network for the 11-year period 1996-2006 based on director data from the Investor Responsibility Research Center director database, and we develop a centrality measure named the influence factor to estimate the influence of directors quantitatively. The US corporate governance network is a network of directors with nodes representing directors and links between two directors representing their service on common company boards. We assume that information flows in the network through information-sharing processes among linked directors. The influence factor assigned to a director is based on the level of information that a director obtains from the entire network. We find that, contrary to commonly accepted belief that directors of large companies, measured by market capitalization, are the most powerful, in some instances, the directors who are influential do not necessarily serve on boards of large companies. By applying our influence factor method to identify the influential people contained in the lists created by popular magazines such as Fortune, Networking World, and Treasury and Risk Management, we find that the influence factor method is consistently either the best or one of the two best methods in identifying powerful people compared to other general centrality measures that are used to denote the significance of a node in complex network theory.

  9. The Potential Use of Wolbachia-Based Mosquito Biocontrol Strategies for Japanese Encephalitis

    PubMed Central

    Jeffries, Claire L.; Walker, Thomas

    2015-01-01

    Japanese encephalitis virus (JEV) is a zoonotic pathogen transmitted by the infectious bite of Culex mosquitoes. The virus causes the development of the disease Japanese encephalitis (JE) in a small proportion of those infected, predominantly affecting children in eastern and southern Asia. Annual JE incidence estimates range from 50,000–175,000, with 25%–30% of cases resulting in mortality. It is estimated that 3 billion people live in countries in which JEV is endemic. The virus exists in an enzootic transmission cycle, with mosquitoes transmitting JEV between birds as reservoir hosts and pigs as amplifying hosts. Zoonotic infection occurs as a result of spillover events from the main transmission cycle. The reservoir avian hosts include cattle egrets, pond herons, and other species of water birds belonging to the family Ardeidae. Irrigated rice fields provide an ideal breeding ground for mosquitoes and attract migratory birds, maintaining the transmission of JEV. Although multiple vaccines have been developed for JEV, they are expensive and require multiple doses to maintain efficacy and immunity. As humans are a “dead-end” host for the virus, vaccination of the human population is unlikely to result in eradication. Therefore, vector control of the principal mosquito vector, Culex tritaeniorhynchus, represents a more promising strategy for reducing transmission. Current vector control strategies include intermittent irrigation of rice fields and space spraying of insecticides during outbreaks. However, Cx. Tritaeniorhynchus is subject to heavy exposure to pesticides in rice fields, and as a result, insecticide resistance has developed. In recent years, significant advancements have been made in the potential use of the bacterial endosymbiont Wolbachia for mosquito biocontrol. The successful transinfection of Wolbachia strains from Drosophila flies to Aedes (Stegomyia) mosquitoes has resulted in the generation of “dengue-refractory” mosquito lines

  10. Cardiovascular emergencies in cruise ship passengers.

    PubMed

    Novaro, Gian M; Bush, Howard S; Fromkin, Kenneth R; Shen, Michael Y; Helguera, Marcelo; Pinski, Sergio L; Asher, Craig R

    2010-01-15

    More than 10 million people, many elderly and likely to harbor cardiovascular (CV) disease, embark on cruise ship travel worldwide every year. The clinical presentation and outcome of CV emergencies presenting during cruise ship travel remain largely unknown. Our department provides contracted cardiology consultations to several large cruise lines. We prospectively maintained a registry of all such consultations during a 2-year period. One hundred consecutive patients were identified (age 66 +/- 14 years, range 18 to 90, 76% men). The most common symptom was chest pain (50%). The most common diagnosis was acute coronary syndrome (58%; ST elevation in 21% and non-ST elevation in 37%). On-board mortality was 3%. Overall, 73% of patients required hospital triage. Of the 25 patients triaged to our institution, 17 underwent a revascularization procedure. One patient died. Ten percent of patients had cardiac symptoms in the days or weeks before boarding; all required hospital triage. Access to a baseline electrocardiogram would have been clinically useful in 23% of cases. In conclusion, CV emergencies, such as acute coronary syndrome and heart failure, are not uncommon on cruise ships. They are often serious, requiring hospital triage and coronary revascularization. A pretravel medical evaluation is recommended for passengers with a cardiac history or a high-risk profile. Passengers should be encouraged to bring a copy of their electrocardiogram on board if abnormal. Cruise lines should establish mechanisms for prompt consultation and triage.

  11. ASPS: American Suborbital Passenger System 2005

    NASA Astrophysics Data System (ADS)

    Wyczalek, Floyd A.

    1994-11-01

    With the evaporation of funding support for the National Aerospace Plane (NASP), realization of a single stage to orbit flight system has been deferred. With the resource down sizing of the NASP management and technical team, development of the NASP concept appears to be delayed beyond the year 2030. To fill this gap for shorter flight times to remote destinations over 8,000 km, it is now proposed to convert the existing NASA shuttle to a passenger configuration and air launch horizontally from a Boeing 747 aircraft as the first stage carrier after climb to launch altitude. This US Shuttle concept may permit the transport of 100 passengers to destinations 10,000 km distant during a flight duration of 90 minutes, and during flight durations of about 120 minutes to any commercial destination on the planet Earth. This long distance system takes advantage of the existing shuttle and Boeing 747 airframe and propulsion technology, and can be implemented within the next decade.

  12. Passenger Deletions Generate Therapeutic Vulnerabilities in Cancer

    PubMed Central

    Muller, Florian L.; Colla, Simona; Aquilanti, Elisa; Manzo, Veronica; Genovese, Giannicola; Lee, Jaclyn; Eisenson, Dan; Narurkar, Rujuta; Deng, Pingna; Nezi, Luigi; Lee, Michelle; Hu, Baoli; Hu, Jian; Sahin, Ergun; Ong, Derrick; Fletcher-Sananikone, Eliot; Ho, Dennis; Kwong, Lawrence; Brennan, Cameron; Wang, Y. Alan; Chin, Lynda; DePinho, Ronald A.

    2013-01-01

    Inactivation of tumor suppressor genes via homozygous deletion is a prototypic event in the cancer genome, yet such deletions often encompass neighboring genes. We hypothesized that homozygous deletions in such passenger genes can expose cancer-specific therapeutic vulnerabilities in the case where the collaterally deleted gene is a member of a functionally redundant family of genes exercising an essential function. The glycolytic gene Enolase 1 (ENO1) in the 1p36 locus is deleted in Glioblastoma (GBM), which is tolerated by expression of ENO2. We demonstrate that shRNA-mediated extinction of ENO2 selectively inhibits growth, survival, and tumorigenic potential of ENO1-deleted GBM cells and that the enolase inhibitor phosphonoacetohydroxamate (PhAH) is selectively toxic to ENO1-deleted GBM cells relative to ENO1-intact GBM cells or normal astrocytes. The principle of collateral vulnerability should be applicable to other passenger deleted genes encoding functionally-redundant essential activities and provide an effective treatment strategy for cancers harboring such genomic events. PMID:22895339

  13. 46 CFR 185.506 - Passenger safety orientation.

    Code of Federal Regulations, 2012 CFR

    2012-10-01

    ... 46 Shipping 7 2012-10-01 2012-10-01 false Passenger safety orientation. 185.506 Section 185.506... TONS) OPERATIONS Preparations for Emergencies § 185.506 Passenger safety orientation. (a) Except as... informed of the required safety information. (e) On a vessel on a voyage of more than 24 hours...

  14. 49 CFR 177.870 - Regulations for passenger carrying vehicles.

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ... be transported by passenger-carrying aircraft or rail car may be transported on a motor vehicle... be transported. (f) Division 6.1 (poisonous) or Division 2.3 (poisonous gas) materials on passenger... Division 2.3 (poisonous gas) material, or any paranitroaniline, in any amount, in or on any bus...

  15. 49 CFR 177.870 - Regulations for passenger carrying vehicles.

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ... be transported by passenger-carrying aircraft or rail car may be transported on a motor vehicle... be transported. (f) Division 6.1 (poisonous) or Division 2.3 (poisonous gas) materials on passenger... Division 2.3 (poisonous gas) material, or any paranitroaniline, in any amount, in or on any bus...

  16. 14 CFR 135.117 - Briefing of passengers before flight.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 14 Aeronautics and Space 3 2010-01-01 2010-01-01 false Briefing of passengers before flight. 135.117 Section 135.117 Aeronautics and Space FEDERAL AVIATION ADMINISTRATION, DEPARTMENT OF... Flight Operations § 135.117 Briefing of passengers before flight. (a) Before each takeoff each pilot...

  17. 14 CFR 135.117 - Briefing of passengers before flight.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ...— (1) Smoking. Each passenger shall be briefed on when, where, and under what conditions smoking is... briefing shall include a statement that the Federal Aviation Regulations require passenger compliance with... designated for safety purposes as no smoking areas, and crewmember instructions with regard to these...

  18. 14 CFR 135.117 - Briefing of passengers before flight.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ...— (1) Smoking. Each passenger shall be briefed on when, where, and under what conditions smoking is... briefing shall include a statement that the Federal Aviation Regulations require passenger compliance with... designated for safety purposes as no smoking areas, and crewmember instructions with regard to these...

  19. 78 FR 71785 - Passenger Train Emergency Systems II

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-11-29

    ...This final rule is intended to further the safety of passenger train occupants through both enhancements and additions to FRA's existing requirements for emergency systems on passenger trains. In this final rule, FRA is adding requirements for emergency passage through vestibule and other interior passageway doors and enhancing emergency egress and rescue access signage requirements. FRA is......

  20. 77 FR 153 - Passenger Train Emergency Systems II

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-01-03

    ...This notice of proposed rulemaking (NPRM) is intended to further the safety of passenger train occupants through both enhancements and additions to FRA's existing requirements for emergency systems on passenger trains. In this NPRM, FRA is proposing to add requirements for interior vestibule doors and enhance emergency egress and rescue access signage requirements. FRA is also proposing to......

  1. 49 CFR 541.5 - Requirements for passenger motor vehicles.

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ...) Engine. (2) Transmission. (3) Right front fender. (4) Left front fender. (5) Hood. (6) Right front door...) Front bumper. (12) Rear bumper. (13) Right rear quarter panel (passenger cars). (14) Left rear quarter panel (passenger cars). (15) Right-side assembly (MPVs). (16) Left-side assembly (MPVs). (17) Pickup...

  2. 75 FR 38878 - Transportation for Individuals With Disabilities: Passenger Vessels

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-07-06

    ...The Department is issuing a new Americans with Disabilities Act (ADA) rule to ensure nondiscrimination on the basis of disability by passenger vessel operators (PVOs). This rulemaking concerns service and policy issues. Issues concerning physical accessibility standards will be addressed at a later time, in conjunction with proposed passenger vessel accessibility guidelines drafted by the......

  3. 27 CFR 31.91 - Passenger trains, aircraft, and vessels.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 27 Alcohol, Tobacco Products and Firearms 1 2011-04-01 2011-04-01 false Passenger trains, aircraft... Sales in Multiple Locations § 31.91 Passenger trains, aircraft, and vessels. Persons who carry on the business of a retail dealer in liquors or of a retail dealer in beer on trains, aircraft, boats, or...

  4. 27 CFR 31.91 - Passenger trains, aircraft, and vessels.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 27 Alcohol, Tobacco Products and Firearms 1 2014-04-01 2014-04-01 false Passenger trains, aircraft... Sales in Multiple Locations § 31.91 Passenger trains, aircraft, and vessels. Persons who carry on the business of a retail dealer in liquors or of a retail dealer in beer on trains, aircraft, boats, or...

  5. 49 CFR 1562.23 - Aircraft operator and passenger requirements.

    Code of Federal Regulations, 2012 CFR

    2012-10-01

    ... 49 Transportation 9 2012-10-01 2012-10-01 false Aircraft operator and passenger requirements. 1562... Certain Operations § 1562.23 Aircraft operator and passenger requirements. (a) General. To operate into or out of DCA, an aircraft operator must: (1) Designate a security coordinator responsible...

  6. 49 CFR 1562.23 - Aircraft operator and passenger requirements.

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ... 49 Transportation 9 2010-10-01 2010-10-01 false Aircraft operator and passenger requirements. 1562... Certain Operations § 1562.23 Aircraft operator and passenger requirements. (a) General. To operate into or out of DCA, an aircraft operator must: (1) Designate a security coordinator responsible...

  7. 49 CFR 1562.23 - Aircraft operator and passenger requirements.

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    ... 49 Transportation 9 2014-10-01 2014-10-01 false Aircraft operator and passenger requirements. 1562... Certain Operations § 1562.23 Aircraft operator and passenger requirements. (a) General. To operate into or out of DCA, an aircraft operator must: (1) Designate a security coordinator responsible...

  8. 49 CFR 1562.23 - Aircraft operator and passenger requirements.

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ... 49 Transportation 9 2013-10-01 2013-10-01 false Aircraft operator and passenger requirements. 1562... Certain Operations § 1562.23 Aircraft operator and passenger requirements. (a) General. To operate into or out of DCA, an aircraft operator must: (1) Designate a security coordinator responsible...

  9. Short haul air passenger data sources in the United States

    NASA Technical Reports Server (NTRS)

    Al-Kazily, J.; Gosling, G.; Horonjeff, R.

    1977-01-01

    The sources and characteristics of existing data on short haul air passenger traffic in the United States domestic air market are described along with data availability, processing, and costs. Reference is made to data derived from aircraft operations since these data can be used to insure that no short haul operators are omitted during the process of assembling passenger data.

  10. 49 CFR 223.8 - Additional requirements for passenger equipment.

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    ... 49 Transportation 4 2014-10-01 2014-10-01 false Additional requirements for passenger equipment. 223.8 Section 223.8 Transportation Other Regulations Relating to Transportation (Continued) FEDERAL RAILROAD ADMINISTRATION, DEPARTMENT OF TRANSPORTATION SAFETY GLAZING STANDARDS-LOCOMOTIVES, PASSENGER...

  11. Passenger and Cell Phone Conversations in Simulated Driving

    ERIC Educational Resources Information Center

    Drews, Frank A.; Pasupathi, Monisha; Strayer, David L.

    2008-01-01

    This study examines how conversing with passengers in a vehicle differs from conversing on a cell phone while driving. We compared how well drivers were able to deal with the demands of driving when conversing on a cell phone, conversing with a passenger, and when driving without any distraction. In the conversation conditions, participants were…

  12. Shuttle passenger couch. [design and performance of engineering model

    NASA Technical Reports Server (NTRS)

    Rosener, A. A.; Stephenson, M. L.

    1974-01-01

    Conceptual design and fabrication of a full scale shuttle passenger couch engineering model are reported. The model was utilized to verify anthropometric dimensions, reach dimensions, ingress/egress, couch operation, storage space, restraint locations, and crew acceptability. These data were then incorported in the design of the passenger couch verification model that underwent performance tests.

  13. 46 CFR 122.502 - Crew and passenger list.

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ... 46 Shipping 4 2010-10-01 2010-10-01 false Crew and passenger list. 122.502 Section 122.502... Preparations for Emergencies § 122.502 Crew and passenger list. (a) The owner, charterer, managing operator, or... Great Lakes port from a Canadian Great Lakes port. (b) The master of a vessel required to prepare a...

  14. Highlighting impact: Do editors' selections identify influential papers?

    NASA Astrophysics Data System (ADS)

    Antonoyiannakis, Manolis

    A recent trend in scientific publishing is that journal editors highlight each week a select set among the papers published (usually) in their respective journals. The highlighted papers are deemed of higher quality, importance, or interest than the 'average' paper and feature prominently in the publishers' websites. We perform a citation analysis of the highlighted papers for a number of journals from various publishers in physics. By comparing the performance of highlighted papers relative to (a) typical papers and (b) highly cited papers in their source journals and in other journals in the field, we explore whether, and to what extent, the selection process at the time of publication identifies papers that will turn out to be influential. We discuss the broader implications for research assessment.

  15. Emergence of Influential Spreaders in Modified Rumor Models

    NASA Astrophysics Data System (ADS)

    Borge-Holthoefer, Javier; Meloni, Sandro; Gonçalves, Bruno; Moreno, Yamir

    2013-04-01

    The burst in the use of online social networks over the last decade has provided evidence that current rumor spreading models miss some fundamental ingredients in order to reproduce how information is disseminated. In particular, recent literature has revealed that these models fail to reproduce the fact that some nodes in a network have an influential role when it comes to spread a piece of information. In this work, we introduce two mechanisms with the aim of filling the gap between theoretical and experimental results. The first model introduces the assumption that spreaders are not always active whereas the second model considers the possibility that an ignorant is not interested in spreading the rumor. In both cases, results from numerical simulations show a higher adhesion to real data than classical rumor spreading models. Our results shed some light on the mechanisms underlying the spreading of information and ideas in large social systems and pave the way for more realistic diffusion models.

  16. Maintenance of adaptive differentiation by Wolbachia induced bidirectional cytoplasmic incompatibility: the importance of sib-mating and genetic systems

    PubMed Central

    Branca, Antoine; Vavre, Fabrice; Silvain, Jean-François; Dupas, Stéphane

    2009-01-01

    Background Bacteria of the genus Wolbachia are reproductive parasites widespread among arthropods. The most common effect arising from the presence of Wolbachia in a population is Cytoplasmic Incompatibility (CI), whereby postmating reproductive isolation occurs in crosses between an infected male and an uninfected female, or when a male is infected with a different strain of Wolbachia to that of the female (bidirectional CI). Previous theoretical models have demonstrated that bidirectional CI can contribute to the genetic divergence of populations in haploid and diploid organisms. However, haplodiploid organisms were not considered in these models even though they include Nasonia parasitoid wasps – the best example of the implication of Wolbachia in ongoing speciation. Moreover, previous work did not investigate inbreeding mating systems, which are frequently observed in arthropod species. Results We developed a stochastic two-island model which simulated three genetic scenarios, diploidy, haploidy, and haplodiploidy, with two CI phenotypes being considered for the latter: (1) male development of female progeny; and (2) mortality of fertilized eggs. We also investigated the effect of varying the proportion of sib mating. In the model each allopatric population was initially fixed for a single allele at a nuclear locus under positive selection and infected with one strain of Wolbachia. Each simulation presupposed that the two populations were fixed for a different allele and a different strain of Wolbachia. The degree of genetic differentiation observed in the locus under selection due to bidirectional CI was much lower for the two haplodiploid phenotypes than for either diploids or haploids. Furthermore, we demonstrated that sib-mating may compensate for the lower efficiency of bidirectional CI in haplodiploids by maintaining genetic divergence. Conclusion Our model suggests that maintenance of genetic differentiation facilitated by Wolbachia is more likely to

  17. Wolbachia and the insect immune system: what reactive oxygen species can tell us about the mechanisms of Wolbachia–host interactions

    PubMed Central

    Zug, Roman; Hammerstein, Peter

    2015-01-01

    Wolbachia are intracellular bacteria that infect a vast range of arthropod species, making them one of the most prevalent endosymbionts in the world. Wolbachia’s stunning evolutionary success is mostly due to their reproductive parasitism but also to mutualistic effects such as increased host fecundity or protection against pathogens. However, the mechanisms underlying Wolbachia phenotypes, both parasitic and mutualistic, are only poorly understood. Moreover, it is unclear how the insect immune system is involved in these phenotypes and why it is not more successful in eliminating the bacteria. Here we argue that reactive oxygen species (ROS) are likely to be key in elucidating these issues. ROS are essential players in the insect immune system, and Wolbachia infection can affect ROS levels in the host. Based on recent findings, we elaborate a hypothesis that considers the different effects of Wolbachia on the oxidative environment in novel vs. native hosts. We propose that newly introduced Wolbachia trigger an immune response and cause oxidative stress, whereas in coevolved symbioses, infection is not associated with oxidative stress, but rather with restored redox homeostasis. Redox homeostasis can be restored in different ways, depending on whether Wolbachia or the host is in charge. This hypothesis offers a mechanistic explanation for several of the observed Wolbachia phenotypes. PMID:26579107

  18. Persistent Infection by Wolbachia wAlbB Has No Effect on Composition of the Gut Microbiota in Adult Female Anopheles stephensi.

    PubMed

    Chen, Shicheng; Zhao, Jiangchao; Joshi, Deepak; Xi, Zhiyong; Norman, Beth; Walker, Edward D

    2016-01-01

    The bacteria in the midgut of Anopheles stephensi adult females from laboratory colonies were studied by sequencing the V4 region of 16S rRNA genes, with respect to three experimental factors: stable or cured Wolbachia infection; sugar or blood diet; and age. Proteobacteria and Bacteroidetes dominated the community [>90% of operational taxonomic units (OTUs)]; most taxa were in the classes Flavobacteriia, Gammaproteobacteria, and Alphaproteobacteria, and were assigned to Elizabethkingia (46.9%), Asaia (6.4%) and Pseudomonas (6.0%), or unclassified Enterobacteriaceae (37.2%). Bacterial communities were similar between Wolbachia-cured and Wolbachia-infected mosquito lines, indicating that the gut microbiota were not dysregulated in the presence of Wolbachia. The proportion of Enterobacteriaceae was higher in mosquitoes fed a blood meal compared to those provided a sugar meal. Collectively, the bacterial community had a similar structure in older Wolbachia-infected mosquitoes 8 days after the blood meal, as in younger Wolbachia-infected mosquitoes before a blood meal, except that older mosquitoes had a higher proportion of Enterobacteriaceae and lower proportion of Elizabethkingia. Consistent presence of certain predominant bacteria (Elizabethkingia, Asaia, Pseudomonas, and Enterobacteriaceae) suggests they would be useful for paratransgenesis to control malaria infection, particularly when coupled to a Wolbachia-based intervention strategy.

  19. Persistent Infection by Wolbachia wAlbB Has No Effect on Composition of the Gut Microbiota in Adult Female Anopheles stephensi

    PubMed Central

    Chen, Shicheng; Zhao, Jiangchao; Joshi, Deepak; Xi, Zhiyong; Norman, Beth; Walker, Edward D.

    2016-01-01

    The bacteria in the midgut of Anopheles stephensi adult females from laboratory colonies were studied by sequencing the V4 region of 16S rRNA genes, with respect to three experimental factors: stable or cured Wolbachia infection; sugar or blood diet; and age. Proteobacteria and Bacteroidetes dominated the community [>90% of operational taxonomic units (OTUs)]; most taxa were in the classes Flavobacteriia, Gammaproteobacteria, and Alphaproteobacteria, and were assigned to Elizabethkingia (46.9%), Asaia (6.4%) and Pseudomonas (6.0%), or unclassified Enterobacteriaceae (37.2%). Bacterial communities were similar between Wolbachia-cured and Wolbachia-infected mosquito lines, indicating that the gut microbiota were not dysregulated in the presence of Wolbachia. The proportion of Enterobacteriaceae was higher in mosquitoes fed a blood meal compared to those provided a sugar meal. Collectively, the bacterial community had a similar structure in older Wolbachia-infected mosquitoes 8 days after the blood meal, as in younger Wolbachia-infected mosquitoes before a blood meal, except that older mosquitoes had a higher proportion of Enterobacteriaceae and lower proportion of Elizabethkingia. Consistent presence of certain predominant bacteria (Elizabethkingia, Asaia, Pseudomonas, and Enterobacteriaceae) suggests they would be useful for paratransgenesis to control malaria infection, particularly when coupled to a Wolbachia-based intervention strategy. PMID:27708633

  20. Emergency medicine and the airline passenger

    NASA Technical Reports Server (NTRS)

    Mohler, S. R.; Nicogossian, A.; Margulies, R. A.

    1980-01-01

    Problems related to immediate medical care in case of in-flight emergencies are discussed with reference to such critical types of medical emergencies as obstructed airway, cardiac dysfunction, trauma, hemorrhage, hypoxia, and pain. It is shown that training flight attendants to deal with in-flight medical emergencies and to use first-aid support equipment and essential and useful drugs may later help with stabilization of a victim and allow continuing the flight to the scheduled destination without the need for a diverted landing. Among the steps suggested in order to upgrade inflight welfare and safety of passengers are the development of an advisory circular by the FAA covering standardized training for flight attendants, regulatory action requiring upgrading of the present rudimentary first-aid kit, and the enactment of Good Samaritan legislation by the U.S. Government.