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Sample records for influential passenger wolbachia

  1. Wolbachia infect ovaries in the course of their maturation: last minute passengers and priority travellers?

    PubMed

    Genty, Lise-Marie; Bouchon, Didier; Raimond, Maryline; Bertaux, Joanne

    2014-01-01

    Wolbachia are widespread endosymbiotic bacteria of arthropods and nematodes. Studies on such models suggest that Wolbachia's remarkable aptitude to infect offspring may rely on a re-infection of ovaries from somatic tissues instead of direct cellular segregation between oogonia and oocytes. In the terrestrial isopod Armadillidium vulgare, Wolbachia are vertically transmitted to the host offspring, even though ovary cells are cyclically renewed. Using Fluorescence in situ hybridization (FISH), we showed that the proportion of infected oocytes increased in the course of ovary and oocyte maturation, starting with 31.5% of infected oocytes only. At the end of ovary maturation, this proportion reached 87.6% for the most mature oocytes, which is close to the known transmission rate to offspring. This enrichment can be explained by a secondary acquisition of the bacteria by oocytes (Wolbachia can be seen as last minute passengers) and/or by a preferential selection of oocytes infected with Wolbachia (as priority travellers). PMID:24722673

  2. Wolbachia Infect Ovaries in the Course of Their Maturation: Last Minute Passengers and Priority Travellers?

    PubMed Central

    Genty, Lise-Marie; Bouchon, Didier; Raimond, Maryline; Bertaux, Joanne

    2014-01-01

    Wolbachia are widespread endosymbiotic bacteria of arthropods and nematodes. Studies on such models suggest that Wolbachia's remarkable aptitude to infect offspring may rely on a re-infection of ovaries from somatic tissues instead of direct cellular segregation between oogonia and oocytes. In the terrestrial isopod Armadillidium vulgare, Wolbachia are vertically transmitted to the host offspring, even though ovary cells are cyclically renewed. Using Fluorescence in situ hybridization (FISH), we showed that the proportion of infected oocytes increased in the course of ovary and oocyte maturation, starting with 31.5% of infected oocytes only. At the end of ovary maturation, this proportion reached 87.6% for the most mature oocytes, which is close to the known transmission rate to offspring. This enrichment can be explained by a secondary acquisition of the bacteria by oocytes (Wolbachia can be seen as last minute passengers) and/or by a preferential selection of oocytes infected with Wolbachia (as priority travellers). PMID:24722673

  3. Horizontal transfers of feminizing versus non-feminizing Wolbachia strains: from harmless passengers to pathogens.

    PubMed

    Le Clec'h, Winka; Raimond, Maryline; Guillot, Sylvain; Bouchon, Didier; Sicard, Mathieu

    2013-11-01

    The endosymbiont Wolbachia pipientis infects various hosts in which it navigates vertically from mothers to offspring. However, horizontal transfers of Wolbachia can occur between hosts. The virulence of the horizontally acquired Wolbachia can change in the new host as it has been illustrated by the case of the feminizing strain wVulC from the woodlouse Armadillidium vulgare that turns to a pathogen when introduced into Porcellio dilatatus dilatatus. In the present study, we aim to show whether symbiotic traits, such as (i) host sex manipulation and (ii) colonization patterns, which differ between eight isopod Wolbachia strains, are connected to their virulence towards the recipient host P. d. dilatatus. Among the transferred Wolbachia, some feminizing strains gradually differing in feminizing intensity in their native hosts induced different levels of pathogenicity to P. d. dilatatus. Not a single feminizing strain passed vertically with high titres to the next generation. The non-feminizing Wolbachia strains, even if they reached high densities in the host, did not impact host life-history traits and some vertically passed with high titres to the offspring. These results suggest that a potential link between the manners Wolbachia manipulates its native host reproduction, its virulence and its ability to vertically infect the offspring. PMID:23802876

  4. 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. PMID:26189661

  5. "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,…

  6. 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

  7. Wolbachia filarial interactions.

    PubMed

    Taylor, Mark J; Voronin, Denis; Johnston, Kelly L; Ford, Louise

    2013-04-01

    Wolbachia pipientis is a widespread intracellular bacterial symbiont of arthropods and is common in insects. One of their more exotic and unexpected hosts is the filarial nematodes, notable for the parasites responsible for onchocerciasis (river blindness), lymphatic filariasis (elephantiasis) and dirofilariasis (heartworm). Wolbachia are only present in a subgroup of the filarial nematodes and do not extend to other groups of nematodes either parasitic or free-living. In the medically and veterinary important species that host Wolbachia, the symbiont has become an essential partner to key biological processes in the life of the nematode to the point where antibiotic elimination of the bacteria leads to a potent and effective anti-filarial drug treatment. We review the cellular and molecular basis of Wolbachia filarial interactions and highlight the key processes provided by the endosymbiont upon which the nematodes have become entirely dependent. This dependency is primarily restricted to periods of the lifecycle with heavy metabolic demands including growth and development of larval stages and embryogenesis in the adult female. Also, the longevity of filarial parasites is compromised following depletion of the symbiont, which for the first time has delivered a safe and effective treatment to kill adult parasites with antibiotics. PMID:23210448

  8. Filarial and Wolbachia genomics.

    PubMed

    Scott, A L; Ghedin, E; Nutman, T B; McReynolds, L A; Poole, C B; Slatko, B E; Foster, J M

    2012-01-01

    Filarial nematode parasites, the causative agents for a spectrum of acute and chronic diseases including lymphatic filariasis and river blindness, threaten the well-being and livelihood of hundreds of millions of people in the developing regions of the world. The 2007 publication on a draft assembly of the 95-Mb genome of the human filarial parasite Brugia malayi- representing the first helminth parasite genome to be sequenced - has been followed in rapid succession by projects that have resulted in the genome sequencing of six additional filarial species, seven nonfilarial nematode parasites of animals and nearly 30 plant parasitic and free-living species. Parallel to the genomic sequencing, transcriptomic and proteomic projects have facilitated genome annotation, expanded our understanding of stage-associated gene expression and provided a first look at the role of epigenetic regulation of filarial genomes through microRNAs. The expansion in filarial genomics will also provide a significant enrichment in our knowledge of the diversity and variability in the genomes of the endosymbiotic bacterium Wolbachia leading to a better understanding of the genetic principles that govern filarial-Wolbachia mutualism. The goal here is to provide an overview of the trends and advances in filarial and Wolbachia genomics. PMID:22098559

  9. Wolbachia versus dengue: Evolutionary forecasts.

    PubMed

    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

  10. 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. PMID:25764472

  11. Uncovering Wolbachia Diversity upon Artificial Host Transfer

    PubMed Central

    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. PMID:24376534

  12. 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. PMID:25710364

  13. Bacteriophage WO in Wolbachia infecting terrestrial isopods.

    PubMed

    Braquart-Varnier, Christine; Grève, Pierre; Félix, Christine; Martin, Gilbert

    2005-11-18

    Wolbachia are maternally inherited intracellular alpha-proteobacteria that infect a wide range of arthropods. They are associated with a number of different reproductive phenotypes in arthropods and nematodes. In isopod crustacean, Wolbachia are responsible for feminization of genetic males in many species, and for cytoplasmic incompatibility in two species. In this paper, we report the first detection of phage WO from Wolbachia infecting terrestrial isopods. All Wolbachia strains tested in this study were infected with phage WO. Based on the orf7 phage sequence, we identified three different phage sequences in four Wolbachia strains. The phage of Wolbachia infecting Armadillidium vulgare seems to be not active, unlike other phages WO previously described in arthropods. PMID:16198306

  14. 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. PMID:26428255

  15. Endosymbiosis: The Remarkable Healing Powers of Wolbachia.

    PubMed

    Sullivan, William

    2016-09-12

    Wolbachia, a maternally transmitted bacterium globally present in arthropods, favors its own transmission by producing dramatic changes in host reproduction. Insight into the underlying molecular and cellular mechanisms comes from the identification of the Wolbachia effector protein TomO, which maintains host germline stem cells in an undifferentiated state. PMID:27623259

  16. Evolution of Wolbachia cytoplasmic incompatibility types.

    PubMed

    Dobson, Stephen L

    2004-10-01

    The success of obligate endosymbiotic Wolbachia infections in insects is due in part to cytoplasmic incompatibility (CI), whereby Wolbachia bacteria manipulate host reproduction to promote their invasion and persistence within insect populations. The observed diversity of CI types raises the question of what the evolutionary pathways are by which a new CI type can evolve from an ancestral type. Prior evolutionary models assume that Wolbachia exists within a host individual as a clonal infection. While endosymbiotic theory predicts a general trend toward clonality, Wolbachia provides an exception in which there is selection to maintain diversity. Here, evolutionary trajectories are discussed that assume that a novel Wolbachia variant will co-exist with the original infection type within a host individual as a superinfection. Relative to prior models, this assumption relaxes requirements and allows additional pathways for the evolution of novel CI types. In addition to describing changes in the Wolbachia infection frequency associated with the hypothesized evolutionary events, the predicted impact of novel CI variants on the host population is also described. This impact, resulting from discordant evolutionary interests of symbiont and host, is discussed as a possible cause of Wolbachia loss from the host population or host population extinction. The latter is also discussed as the basis for an applied strategy for the suppression of insect pest populations. Model predictions are discussed relative to a recently published Wolbachia genome sequence and prior characterization of CI in naturally and artificially infected insects. PMID:15562682

  17. Wolbachia-Host Interactions: Host Mating Patterns Affect Wolbachia Density Dynamics

    PubMed Central

    Zhao, Dong-Xiao; Zhang, Xiang-Fei; Chen, Da-Song; Zhang, Yan-Kai; Hong, Xiao-Yue

    2013-01-01

    Wolbachia are maternally inherited intracellular bacteria that infect a wide range of arthropods and cause an array of effects on host reproduction, fitness and mating behavior. Although our understanding of the Wolbachia-associated effects on hosts is rapidly expanding, our knowledge of the host factors that mediate Wolbachia dynamics is rudimentary. Here, we explore the interactions between Wolbachia and its host, the two-spotted spider mite Tetranychus urticae Koch. Our results indicate that Wolbachia induces strong cytoplasmic incompatibility (CI), increases host fecundity, but has no effects on the longevity of females and the mating competitiveness of males in T. urticae. Most importantly, host mating pattern was found to affect Wolbachia density dynamics during host aging. Mating of an uninfected mite of either sex with an infected mite attenuates the Wolbachia density in the infected mite. According to the results of Wolbachia localization, this finding may be associated with the tropism of Wolbachia for the reproductive tissue in adult spider mites. Our findings describe a new interaction between Wolbachia and their hosts. PMID:23823081

  18. 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. PMID:25046729

  19. Iron necessity: the secret of Wolbachia's success?

    PubMed

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

    2014-10-01

    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

  20. 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

  1. Wolbachia in Anastrepha fruit flies (Diptera: Tephritidae).

    PubMed

    Coscrato, Virginia E; Braz, Antônio S K; P Perondini, André L; Selivon, Denise; Marino, Celso L

    2009-09-01

    Endosymbiotic bacteria of the genus Wolbachia are widespread among arthropods and cause a variety of reproductive abnormalities, such as cytoplasmic incompatibility, thelytokous parthenogenesis, male-killing, and host feminization. In this study, we used three sets of Wolbachia-specific primers (16S rDNA, ftsZ, and wsp) in conjunction with the polymerase chain reaction (PCR), cloning and sequencing to study the infection of fruit flies (Anastrepha spp. and Ceratitis capitata) by Wolbachia. The flies were collected at several localities in Brazil and at Guayaquil, Ecuador. All of the fruit flies studied were infected with Wolbachia supergroup A, in agreement with the high prevalence of this group in South America. Phylogenetic analysis showed that the wsp gene was the most sensitive gene for studying the relationships among Wolbachia strains. The Wolbachia sequences detected in these fruit flies were similar to those such as wMel reported for other fruit flies. These results show that the infection of Anastrepha fruit flies by Wolbachia is much more widespread than previously thought. PMID:19536597

  2. 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...

  3. Multiple Rescue Factors Within a Wolbachia Strain

    PubMed Central

    Zabalou, Sofia; Apostolaki, Angeliki; Pattas, Savvas; Veneti, Zoe; Paraskevopoulos, Charalampos; Livadaras, Ioannis; Markakis, George; Brissac, Terry; Merçot, Hervé; Bourtzis, Kostas

    2008-01-01

    Wolbachia-induced cytoplasmic incompatibility (CI) is expressed when infected males are crossed with either uninfected females or females infected with Wolbachia of different CI specificity. In diploid insects, CI results in embryonic mortality, apparently due to the the loss of the paternal set of chromosomes, usually during the first mitotic division. The molecular basis of CI has not been determined yet; however, several lines of evidence suggest that Wolbachia exhibits two distinct sex-dependent functions: in males, Wolbachia somehow “imprints” the paternal chromosomes during spermatogenesis (mod function), whereas in females, the presence of the same Wolbachia strain(s) is able to restore embryonic viability (resc function). On the basis of the ability of Wolbachia to induce the modification and/or rescue functions in a given host, each bacterial strain can be classified as belonging in one of the four following categories: mod+ resc+, mod− resc+, mod− resc−, and mod+ resc−. A so-called “suicide” mod+ resc− strain has not been found in nature yet. Here, a combination of embryonic cytoplasmic injections and introgression experiments was used to transfer nine evolutionary, distantly related Wolbachia strains (wYak, wTei, wSan, wRi, wMel, wHa, wAu, wNo, and wMa) into the same host background, that of Drosophila simulans (STCP strain), a highly permissive host for CI expression. We initially characterized the modification and rescue properties of the Wolbachia strains wYak, wTei, and wSan, naturally present in the yakuba complex, upon their transfer into D. simulans. Confocal microscopy and multilocus sequencing typing (MLST) analysis were also employed for the evaluation of the CI properties. We also tested the compatibility relationships of wYak, wTei, and wSan with all other Wolbachia infections. So far, the cytoplasmic incompatibility properties of different Wolbachia variants are explained assuming a single pair of modification and rescue

  4. Dietary Cholesterol Modulates Pathogen Blocking by Wolbachia

    PubMed Central

    Caragata, Eric P.; Rancès, Edwige; Hedges, Lauren M.; Gofton, Alexander W.; Johnson, Karyn N.; O'Neill, Scott L.; McGraw, Elizabeth A.

    2013-01-01

    The bacterial endosymbiont Wolbachia pipientis protects its hosts from a range of pathogens by limiting their ability to form infections inside the insect. This “pathogen blocking” could be explained by innate immune priming by the symbiont, competition for host-derived resources between pathogens and Wolbachia, or the direct modification of the cell or cellular environment by Wolbachia. Recent comparative work in Drosophila and the mosquito Aedes aegypti has shown that an immune response is not required for pathogen blocking, implying that there must be an additional component to the mechanism. Here we have examined the involvement of cholesterol in pathogen blocking using a system of dietary manipulation in Drosophila melanogaster in combination with challenge by Drosophila C virus (DCV), a common fly pathogen. We observed that flies reared on cholesterol-enriched diets infected with the Wolbachia strains wMelPop and wMelCS exhibited reduced pathogen blocking, with viral-induced mortality occurring 2–5 days earlier than flies reared on Standard diet. This shift toward greater virulence in the presence of cholesterol also corresponded to higher viral copy numbers in the host. Interestingly, an increase in dietary cholesterol did not have an effect on Wolbachia density except in one case, but this did not directly affect the strength of pathogen blocking. Our results indicate that host cholesterol levels are involved with the ability of Wolbachia-infected flies to resist DCV infections, suggesting that cholesterol contributes to the underlying mechanism of pathogen blocking. PMID:23825950

  5. The Mutualistic Side of Wolbachia-Isopod Interactions: Wolbachia Mediated Protection Against Pathogenic Intracellular Bacteria.

    PubMed

    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

  6. 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. PMID:25921489

  7. Detection and characterization of Wolbachia infection in silkworm

    PubMed Central

    Zha, Xingfu; Zhang, Wenji; Zhou, Chunyan; Zhang, Liying; Xiang, Zhonghuai; Xia, Qingyou

    2014-01-01

    Wolbachia naturally infects a wide variety of arthropods, where it plays important roles in host reproduction. It was previously reported that Wolbachia did not infect silkworm. By means of PCR and sequencing we found in this study that Wolbachia is indeed present in silkworm. Phylogenetic analysis indicates that Wolbachia infection in silkworm may have occurred via transfer from parasitic wasps. Furthermore, Southern blotting results suggest a lateral transfer of the wsp gene into the genomes of some wild silkworms. By antibiotic treatments, we found that tetracycline and ciprofloxacin can eliminate Wolbachia in the silkworm and Wolbachia is important to ovary development of silkworm. These results provide clues towards a more comprehensive understanding of the interaction between Wolbachia and silkworm and possibly other lepidopteran insects. PMID:25249781

  8. 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

  9. Mosaic nature of the wolbachia surface protein.

    PubMed

    Baldo, Laura; Lo, Nathan; Werren, John H

    2005-08-01

    Lateral gene transfer and recombination play important roles in the evolution of many parasitic bacteria. Here we investigate intragenic recombination in Wolbachia bacteria, considered among the most abundant intracellular bacteria on earth. We conduct a detailed analysis of the patterns of variation and recombination within the Wolbachia surface protein, utilizing an extensive set of published and new sequences from five main supergroups of Wolbachia. Analysis of nucleotide and amino acid sequence variations confirms four hypervariable regions (HVRs), separated by regions under strong conservation. Comparison of shared polymorphisms reveals a complex mosaic structure of the gene, characterized by a clear intragenic recombining of segments among several distinct strains, whose major recombination effect is shuffling of a relatively conserved set of amino acid motifs within each of the four HVRs. Exchanges occurred both within and between the arthropod supergroups. Analyses based on phylogenetic methods and a specific recombination detection program (MAXCHI) significantly support this complex partitioning of the gene, indicating a chimeric origin of wsp. Although wsp has been widely used to define macro- and microtaxonomy among Wolbachia strains, these results clearly show that it is not suitable for this purpose. The role of wsp in bacterium-host interactions is currently unknown, but results presented here indicate that exchanges of HVR motifs are favored by natural selection. Identifying host proteins that interact with wsp variants should help reveal how these widespread bacterial parasites affect and evolve in response to the cellular environments of their invertebrate hosts. PMID:16030235

  10. 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

  11. Population Genomics of the Wolbachia Endosymbiont in Drosophila melanogaster

    PubMed Central

    Richardson, Mark F.; Weinert, Lucy A.; Welch, John J.; Linheiro, Raquel S.; Magwire, Michael M.; Jiggins, Francis M.; Bergman, Casey M.

    2012-01-01

    Wolbachia are maternally inherited symbiotic bacteria, commonly found in arthropods, which are able to manipulate the reproduction of their host in order to maximise their transmission. The evolutionary history of endosymbionts like Wolbachia can be revealed by integrating information on infection status in natural populations with patterns of sequence variation in Wolbachia and host mitochondrial genomes. Here we use whole-genome resequencing data from 290 lines of Drosophila melanogaster from North America, Europe, and Africa to predict Wolbachia infection status, estimate relative cytoplasmic genome copy number, and reconstruct Wolbachia and mitochondrial genome sequences. Overall, 63% of Drosophila strains were predicted to be infected with Wolbachia by our in silico analysis pipeline, which shows 99% concordance with infection status determined by diagnostic PCR. Complete Wolbachia and mitochondrial genomes show congruent phylogenies, consistent with strict vertical transmission through the maternal cytoplasm and imperfect transmission of Wolbachia. Bayesian phylogenetic analysis reveals that the most recent common ancestor of all Wolbachia and mitochondrial genomes in D. melanogaster dates to around 8,000 years ago. We find evidence for a recent global replacement of ancestral Wolbachia and mtDNA lineages, but our data suggest that the derived wMel lineage arose several thousand years ago, not in the 20th century as previously proposed. Our data also provide evidence that this global replacement event is incomplete and is likely to be one of several similar incomplete replacement events that have occurred since the out-of-Africa migration that allowed D. melanogaster to colonize worldwide habitats. This study provides a complete genomic analysis of the evolutionary mode and temporal dynamics of the D. melanogaster–Wolbachia symbiosis, as well as important resources for further analyses of the impact of Wolbachia on host biology. PMID:23284297

  12. 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. PMID:25899523

  13. 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

  14. A feedback loop between Wolbachia and the Drosophila gurken mRNP complex influences Wolbachia titer

    PubMed Central

    Serbus, Laura R.; Ferreccio, Amy; Zhukova, Mariya; McMorris, Chanel L.; Kiseleva, Elena; Sullivan, William

    2011-01-01

    Although much is known about interactions between bacterial endosymbionts and their hosts, little is known concerning the host factors that influence endosymbiont titer. Wolbachia endosymbionts are globally dispersed throughout most insect species and are the causative agent in filarial nematode-mediated disease. Our investigation indicates that gurken (grk), a host gene encoding a crucial axis determinant, has a cumulative, dosage-sensitive impact on Wolbachia growth and proliferation during Drosophila oogenesis. This effect appears to be mediated by grk mRNA and its protein-binding partners Squid and Hrp48/Hrb27C, implicating the grk mRNA–protein (mRNP) complex as a rate-limiting host factor controlling Wolbachia titer. Furthermore, highly infected flies exhibit defects that match those occurring with disruption of grk mRNPs, such as nurse cell chromatin disruptions and malformation of chorionic appendages. These findings suggest a feedback loop in which Wolbachia interaction with the grk mRNP affects both Wolbachia titer and grk mRNP function. PMID:22193955

  15. Native microbiome impedes vertical transmission of Wolbachia in Anopheles mosquitoes

    PubMed Central

    Hughes, Grant L.; Dodson, Brittany L.; Johnson, Rebecca M.; Murdock, Courtney C.; Tsujimoto, Hitoshi; Suzuki, Yasutsugu; Patt, Alyssa A.; Cui, Long; Nossa, Carlos W.; Barry, Rhiannon M.; Sakamoto, Joyce M.; Hornett, Emily A.; Rasgon, Jason L.

    2014-01-01

    Over evolutionary time, Wolbachia has been repeatedly transferred between host species contributing to the widespread distribution of the symbiont in arthropods. For novel infections to be maintained, Wolbachia must infect the female germ line after being acquired by horizontal transfer. Although mechanistic examples of horizontal transfer exist, there is a poor understanding of factors that lead to successful vertical maintenance of the acquired infection. Using Anopheles mosquitoes (which are naturally uninfected by Wolbachia) we demonstrate that the native mosquito microbiota is a major barrier to vertical transmission of a horizontally acquired Wolbachia infection. After injection into adult Anopheles gambiae, some strains of Wolbachia invade the germ line, but are poorly transmitted to the next generation. In Anopheles stephensi, Wolbachia infection elicited massive blood meal-induced mortality, preventing development of progeny. Manipulation of the mosquito microbiota by antibiotic treatment resulted in perfect maternal transmission at significantly elevated titers of the wAlbB Wolbachia strain in A. gambiae, and alleviated blood meal-induced mortality in A. stephensi enabling production of Wolbachia-infected offspring. Microbiome analysis using high-throughput sequencing identified that the bacterium Asaia was significantly reduced by antibiotic treatment in both mosquito species. Supplementation of an antibiotic-resistant mutant of Asaia to antibiotic-treated mosquitoes completely inhibited Wolbachia transmission and partly contributed to blood meal-induced mortality. These data suggest that the components of the native mosquito microbiota can impede Wolbachia transmission in Anopheles. Incompatibility between the microbiota and Wolbachia may in part explain why some hosts are uninfected by this endosymbiont in nature. PMID:25114252

  16. Comparative Genomics of a Parthenogenesis-Inducing Wolbachia Symbiont.

    PubMed

    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. 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

  18. Macronutrients mediate the functional relationship between Drosophila and Wolbachia

    PubMed Central

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

    2015-01-01

    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. PMID:25520356

  19. The Impact of Host Diet on Wolbachia Titer in Drosophila

    PubMed Central

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

    2015-01-01

    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. PMID:25826386

  20. 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

  1. 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

  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. PMID:26664683

  3. 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

  4. Monitoring long-term evolutionary changes following Wolbachia introduction into a novel host: the Wolbachia popcorn infection in Drosophila simulans

    PubMed Central

    Carrington, Lauren B.; Hoffmann, Ary A.; Weeks, Andrew R.

    2010-01-01

    Wolbachia may act as a biological control agent for pest management; in particular, the Wolbachia variant wMelPop (popcorn) shortens host longevity and may be useful for dengue suppression. However, long-term changes in the host and Wolbachia genomes can alter Wolbachia spread and/or host effects that suppress disease. Here, we investigate the phenotypic effects of wMelPop in a non-native host, Drosophila simulans, following artificial transinfection approximately 200 generations ago. Long-term rearing and maintenance of the bacteria were at 19°C in the original I-102 genetic background that was transinfected with the popcorn strain. The bacteria were then introgressed into three massbred backgrounds, and tetracycline was used to create uninfected sublines. The effect of wMelPop on longevity in this species appears to have changed; longevity was no longer reduced at 25°C in some nuclear backgrounds, reflecting different geographical origin, selection or drift, although the reduction was still evident for flies held at 30°C. Wolbachia influenced productivity and viability, and development time in some host backgrounds. These findings suggest that long-term attenuation of Wolbachia effects may compromise the effectiveness of this bacterium in pest control. They also emphasize the importance of host nuclear background on Wolbachia phenotypic effects. PMID:20219740

  5. Wolbachia infection shared among planthoppers (Homoptera: Delphacidae) and their endoparasite (Strepsiptera: Elenchidae): a probable case of interspecies transmission.

    PubMed

    Noda, H; Miyoshi, T; Zhang, Q; Watanabe, K; Deng, K; Hoshizaki, S

    2001-08-01

    Wolbachia, a group of parasitic bacteria of arthropods, are believed to be horizontally transmitted among arthropod taxa. We present a new probable example of interspecies horizontal transmission of Wolbachia by way of an endoparasite based on the conformity of Wolbachia gene sequences. Field samples of two rice planthoppers, Laodelphax striatellus and Sogatella furcifera possessed identical Wolbachia. Among three major endoparasites of planthoppers, a strepsipteran, Elenchus japonicus, harboured the identical Wolbachia strain, suggesting strepsipteran transmission of Wolbachia from one planthopper to the other. No Wolbachia was detected in a mermithid nematode Agamermis unka, and dryinid wasps possessed different types of Wolbachia. PMID:11555254

  6. 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.

  7. 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. PMID:26776329

  8. Wolbachia infection lowers fertile sperm transfer in a moth

    PubMed Central

    Lewis, Z.; Champion de Crespigny, F. E.; Sait, S. M.; Tregenza, T.; Wedell, N.

    2011-01-01

    The endosymbiotic bacterium Wolbachia pipientis manipulates host reproduction by rendering infected males reproductively incompatible with uninfected females (cytoplasmic incompatibility; CI). CI is believed to occur as a result of Wolbachia-induced modifications to sperm during maturation, which prevent infected sperm from initiating successful zygote development when fertilizing uninfected females' eggs. However, the mechanism by which CI occurs has been little studied outside the genus Drosophila. Here, we show that in the sperm heteromorphic Mediterranean flour moth, Ephestia kuehniella, infected males transfer fewer fertile sperm at mating than uninfected males. In contrast, non-fertile apyrene sperm are not affected. This indicates that Wolbachia may only affect fertile sperm production and highlights the potential of the Lepidoptera as a model for examining the mechanism by which Wolbachia induces CI in insects. PMID:20880864

  9. Interactions between coexisting intracellular genomes: mitochondrial density and Wolbachia infection.

    PubMed

    Mouton, L; Henri, H; Fleury, F

    2009-04-01

    Many arthropods are infected with maternally transmitted microorganisms, leading to the coexistence of several intracellular genomes within the host cells, including their own mitochondria. As these genomes are cotransmitted, their patterns of evolution have been intimately linked, with possible consequences for the diversity and evolution of the host mitochondrial DNA. The evolutionary aspects of the situation have been thoroughly investigated, especially the selective sweep on the mitochondria as a result of Wolbachia invasion, whereas direct interactions between mitochondria and intracellular symbionts within the host cells or body have received little attention. Since endosymbionts exploit host resources but mitochondria supply energy to meet the bioenergetic demands of organisms, an unanswered question concerns the correlation between their densities. Here, we investigated the influence of Wolbachia symbiosis on mitochondrial density in two parasitic wasps of Drosophila species, both of which are naturally infected by three Wolbachia strains, but they differ in their degree of dependency on these bacteria. In Leptopilina heterotoma, all Wolbachia strains are facultative, whereas Asobara tabida requires a strain of Wolbachia for oogenesis to occur. In both species, Wolbachia infections are stable and well regulated, since the density of each strain does not depend on the presence or absence of other strains. Using lines that harbor various Wolbachia infection statuses, we found that mitochondrial density was not affected by the infection regardless of the sex and age of the host, which is strongly reminiscent of the independent regulation of specific Wolbachia strains and suggest that the protagonists coexist independently of each other as the result of a long-term coevolutionary interaction. PMID:19181828

  10. 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

  11. Addition of wsp sequences to the Wolbachia phylogenetic tree and stability of the classification.

    PubMed

    Pintureau, B; Chaudier, S; Lassablière, F; Charles, H; Grenier, S

    2000-10-01

    Wolbachia are symbiotic bacteria altering reproductive characters of numerous arthropods. Their most recent phylogeny and classification are based on sequences of the wsp gene. We sequenced wsp gene from six Wolbachia strains infecting six Trichogramma species that live as egg parasitoids on many insects. This allows us to test the effect of the addition of sequences on the Wolbachia phylogeny and to check the classification of Wolbachia infecting Trichogramma. The six Wolbachia studied are classified in the B supergroup. They confirm the monophyletic structure of the B Wolbachia in Trichogramma but introduce small differences in the Wolbachia classification. Modifications include the definition of a new group, Sem, for Wolbachia of T. semblidis and the merging of the two closely related groups, Sib and Kay. Specific primers were determined and tested for the Sem group. PMID:11040288

  12. 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. PMID:25259690

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

    PubMed

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

    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

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

    PubMed

    Lefoulon, Emilie; Bain, Odile; Makepeace, Benjamin L; d'Haese, Cyrille; Uni, Shigehiko; Martin, Coralie; 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

  15. 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

  16. 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

  17. Integral aircraft passenger seat

    NASA Technical Reports Server (NTRS)

    Kubokawa, C. C.

    1973-01-01

    Human-engineering approach was used to design integral seat which provides all the safety, comfort, and protective features that can possibly be afforded airline passengers. Results of dynamic impact testing indicated that seat can withstand and attenuate gravity loads of 21-g horizontal and 45-g vertical; by design, seat will withstand lateral g's as well.

  18. 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. PMID:24909063

  19. 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. PMID:25541519

  20. Wolbachia Infection in a Natural Parasitoid Wasp Population

    PubMed Central

    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. PMID:26244782

  1. Phylogeny and PCR-based classification of Wolbachia strains using wsp gene sequences.

    PubMed Central

    Zhou, W; Rousset, F; O'Neil, S

    1998-01-01

    Wolbachia are a group of intracellular inherited bacteria that infect a wide range of arthropods. They are associated with a number of different reproductive phenotypes in their hosts, such as cytoplasmic incompatibility, parthenogenesis and feminization. While it is known that the bacterial strains responsible for these different host phenotypes form a single clade within the alpha-Proteobacteria, until now it has not been possible to resolve the evolutionary relationships between different Wolbachia strains. To address this issue we have cloned and sequenced a gene encoding a surface protein of Wolbachia (wsp) from a representative sample of 28 Wolbachia strains. The sequences from this gene were highly variable and could be used to resolve the phylogenetic relationships of different Wolbachia strains. Based on the sequence of the wsp gene from different Wolbachia isolates we propose that the Wolbachia pipientis clade be initially divided into 12 groups. As more sequence information becomes available we expect the number of such groups to increase. In addition, we present a method of Wolbachia classification based on the use of group-specific wsp polymerase chain reaction (PGR) primers which will allow Wolbachia isolates to be typed without the need to clone and sequence individual Wolbachia genes. This system should facilitate future studies investigating the distribution and biology of Wolbachia strains from large samples of different host species. PMID:9569669

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

    PubMed Central

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

    2014-01-01

    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. PMID:24905191

  3. 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

  4. 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

  5. Phage WO of Wolbachia: lambda of the endosymbiont world

    PubMed Central

    Kent, Bethany N.; Bordenstein, Seth R.

    2010-01-01

    The discovery of an extraordinarily high level of mobile elements in the genome of Wolbachia, a widespread arthropod and nematode endosymbiont, suggests that this bacterium could be an excellent model for assessing the evolution and function of mobile DNA in specialized bacteria. Here, we discuss how studies on the temperate bacteriophage WO of Wolbachia have revealed unexpected levels of genomic flux and are challenging previously held views about the clonality of obligate intracellular bacteria. We also discuss the roles that this phage might play in the Wolbachia-arthropod symbiosis, and infer how this research can be translated to combating human diseases vectored by arthropods. We expect that this temperate phage will be a preeminent model system to understand phage genetics, evolution, and ecology in obligate intracellular bacteria. In this sense, phage WO might be likened to phage λ of the endosymbiont world. PMID:20083406

  6. 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. PMID:21422088

  7. 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. PMID:21422094

  8. 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

  9. Passage of Wolbachia pipientis through Mutant Drosophila melanogaster Induces Phenotypic and Genomic Changes

    PubMed Central

    Sheehan, Kathy B.

    2014-01-01

    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. PMID:25452279

  10. 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

  11. 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

  12. Energy efficient passenger vehicle

    SciTech Connect

    Dessert, R.

    1983-02-22

    An energy efficient passenger carrying vehicle for road use. The vehicle basically comprises a long, narrow body carrying two passengers in a back-to-back relationship. The vehicle is basically a battery powered electric vehicle that can be charged by all free energy sources; namely, the sun, the wind, human muscles and momentum. The vehicle comprises four modules, namely body, solar, and two power modules. An electric power module is located within each end of the body module. This module includes electric motors driving the vehicle supporting wheels and rechargeable batteries to power the motors. Pedals, similar to those on a bicycle, located at each power module, drive generators to help recharge the batteries during operation of the vehicle, or directly help drive the vehicle wheels. A solar module comprising a large electricity generating solar cell panel covers most of the vehicle roof to aid in charging the batteries. Means are provided to tilt the solar cell panel toward the sun about a longitudinal axis. A unique flexible duct below the solar panel serves to cool the cells and, if desired, heat the passenger compartment. Further energy savings are obtained by canting the rear wheels while steering with the front wheels, so that the vehicle moves down the road at a crab angle which provides a sail effect when wind is from the vehicle beam or aft of the beam. Regenerative braking means can be used when slowing down, on a long down grade, when sailing speed is greater than required, or any other time when vehicle momentum is greater than necessary for vehicle operation, to use the excess forward momentum to drive generators to charge the batteries. Thus, a single battery charge will be conserved and vehicle operation will be assisted in a manner giving maximum vehicle range and speed.

  13. Energy efficient passenger vehicle

    SciTech Connect

    Dessert, R.

    1980-01-01

    An energy efficient passenger carrying vehicle for road use comprised of a long, narrow body carrying two passengers in a back-to-back relationship is described. The vehicle is basically a battery powered electric vehicle that can be charged by all free energy sources; namely, the sun, the wind, human muscles and momentum. The vehicle comprises four modules: body, solar, and two power modules. An electric power module is located within each end of the body module. This module includes electric motors driving the vehicle supporting wheels and rechargeable batteries to power the motors. Pedals, similar to those on a bicycle, located at each power module, drive generators to help recharge the batteries during operation of the vehicle, or directly help drive the vehicle wheels. A solar module comprising a large electricity generating solar cell panel covers most of the vehicle roof to aid in charging the batteries. Means are provided to tilt the solar cell panel toward the sun about a longitudinal axis. A unique flexible duct below the solar panel serves to cool the cells and, if desired, heat the passenger compartment. Further energy savings are obtained by canting the rear wheels while steering with the front wheels, so that the vehicle moves down the road at a crab angle which provides a sail effect when wind is from the vehicle beam or aft of the beam. Regenerative braking means can be used when slowing down, on a long down grade, when sailing speed is greater than required, or any other time when vehicle momentum is greater than necessary for vehicle operation, to use the excess forward momentum to drive generators to charge the batteries. Thus, a single battery charge will be conserved and vehicle operation will be assisted in a manner giving maximum vehicle range and speed.

  14. MULTIPLE STRAIN WOLBACHIA INFECTION OF NORTHERN CORN ROOTWORM (DIABROTICA BARBERI)

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Northern corn rootworm (Diabrotica barberi)(NCR) populations in the USA are infected with at least 3 strains of the endosymbiont, Wolbachia. NCR from central Illinois appears to be singly infected with a strain wBar2. NCR from eastern Illinois to Pennsylvania appear to harbor at least 2 differen...

  15. 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. PMID:26051887

  16. The Diversity and Evolution of Wolbachia Ankyrin Repeat Domain Genes

    PubMed Central

    Siozios, Stefanos; Ioannidis, Panagiotis; Klasson, Lisa; Andersson, Siv G. E.; Braig, Henk R.; Bourtzis, Kostas

    2013-01-01

    Ankyrin repeat domain-encoding genes are common in the eukaryotic and viral domains of life, but they are rare in bacteria, the exception being a few obligate or facultative intracellular Proteobacteria species. Despite having a reduced genome, the arthropod strains of the alphaproteobacterium Wolbachia contain an unusually high number of ankyrin repeat domain-encoding genes ranging from 23 in wMel to 60 in wPip strain. This group of genes has attracted considerable attention for their astonishing large number as well as for the fact that ankyrin proteins are known to participate in protein-protein interactions, suggesting that they play a critical role in the molecular mechanism that determines host-Wolbachia symbiotic interactions. We present a comparative evolutionary analysis of the wMel-related ankyrin repeat domain-encoding genes present in different Drosophila-Wolbachia associations. Our results show that the ankyrin repeat domain-encoding genes change in size by expansion and contraction mediated by short directly repeated sequences. We provide examples of intra-genic recombination events and show that these genes are likely to be horizontally transferred between strains with the aid of bacteriophages. These results confirm previous findings that the Wolbachia genomes are evolutionary mosaics and illustrate the potential that these bacteria have to generate diversity in proteins potentially involved in the symbiotic interactions. PMID:23390535

  17. Assessing the epidemiological impact of Wolbachia deployment for dengue control

    PubMed Central

    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.

    2016-01-01

    Summary Dengue viruses cause more human morbidity and mortality than any other arthropod-borne virus. Dengue prevention relies primarily on vector control but 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 half a century ago, it has only gained significant interest as a potential agent of dengue control in the last decade. Here, we review the evidence that supports a practical approach for dengue reduction through field release of Wolbachia-infected mosquitoes and discuss the additional studies that must be conducted before the strategy can be validated and operationally implemented. A critical next step is to assess the efficacy of Wolbachia deployment in reducing dengue virus transmission. We argue that a cluster-randomized trial is currently premature because Wolbachia strain choice for release as well as deployment strategies are still being optimized. We therefore present a pragmatic approach to acquiring preliminary evidence of efficacy via a suite of complementary methodologies: prospective cohort study, geographical cluster investigation, virus phylogenetic analysis, virus surveillance in mosquitoes, and vector competence assays. This multi-pronged approach could provide valuable intermediate evidence of efficacy to justify a future cluster-randomized trial. PMID:26051887

  18. 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

  19. 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. PMID:26579107

  20. Wolbachia is not all about sex: male-feminizing Wolbachia alters the leafhopper Zyginidia pullula transcriptome in a mainly sex-independent manner

    PubMed Central

    Asgharian, Hosseinali; Chang, Peter L.; Mazzoglio, Peter J.; Negri, Ilaria

    2014-01-01

    Wolbachia causes the feminization of chromosomally male embryos in several species of crustaceans and insects, including the leafhopper Zyginidia pullula. In contrast to the relatively well-established ecological aspects of male feminization (e.g., sex ratio distortion and its consequences), the underlying molecular mechanisms remain understudied and unclear. We embarked on an exploratory study to investigate the extent and nature of Wolbachia's effect on gene expression pattern in Z. pullula. We sequenced whole transcriptomes from Wolbachia-infected and uninfected adults. 18147 loci were assembled de novo, including homologs of several Drosophila sex determination genes. A number of transcripts were flagged as candidate Wolbachia sequences. Despite the resemblance of Wolbachia-infected chromosomal males to uninfected and infected chromosomal females in terms of sexual morphology and behavior, principal component analysis revealed that gene expression patterns did not follow these sexual phenotype categories. The principal components generated by differentially expressed genes specified a strong sex-independent Wolbachia effect, followed by a weaker Wolbachia-sexual karyotype interaction effect. Approaches to further examine the molecular mechanism of Wolbachia-host interactions have been suggested based on the presented findings. PMID:25225494

  1. 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…

  2. Wolbachia-induced cytoplasmic incompatibility as a means for insect pest population control

    PubMed Central

    Zabalou, Sofia; Riegler, Markus; Theodorakopoulou, Marianna; Stauffer, Christian; Savakis, Charalambos; Bourtzis, Kostas

    2004-01-01

    Biological control is the purposeful introduction of parasites, predators, and pathogens to reduce or suppress pest populations. Wolbachia are inherited bacteria of arthropods that have recently attracted attention for their potential as new biocontrol agents. Wolbachia manipulate host reproduction by using several strategies, one of which is cytoplasmic incompatibility (CI) [Stouthamer, R., Breeuwer, J. A. J. & Hurst, G. D. D. (1999) Annu. Rev. Microbiol. 53, 71–102]. We established Wolbachia-infected lines of the medfly Ceratitis capitata using the infected cherry fruit fly Rhagoletis cerasi as donor. Wolbachia induced complete CI in the novel host. Laboratory cage populations were completely suppressed by single releases of infected males, suggesting that Wolbachia-induced CI could be used as a novel environmentally friendly tool for the control of medfly populations. The results also encourage the introduction of Wolbachia into pest and vector species of economic and hygenic relevance to suppress or modify natural populations. PMID:15469918

  3. Interaction between host genotype and environmental conditions affects bacterial density in Wolbachia symbiosis.

    PubMed

    Mouton, Laurence; Henri, Hélène; Charif, Delphine; Boulétreau, Michel; Vavre, Fabrice

    2007-04-22

    Regulation of microbial population density is a necessity in stable symbiotic interactions. In Wolbachia symbiosis, both bacterial and host genotypes are involved in density regulation, but environmental factors may also affect bacterial population density. Here, we studied the interaction between three strains of Wolbachia in two divergent homozygous lines of the wasp Leptopilina heterotoma at two different temperatures. Wolbachia density varied between the two host genotypes at only one temperature. Moreover, at this temperature, reciprocal-cross F1 insects displayed identical Wolbachia densities, which were intermediate between the densities in the two parental lines. While these findings confirm that the host genotype plays an important role in Wolbachia density, they also highlight its interaction with environmental conditions, making possible the evolution of local adaptations for the regulation of Wolbachia density. PMID:17251124

  4. 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

  5. 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 ...

  6. No effect of Wolbachia on resistance to intracellular infection by pathogenic bacteria in Drosophila melanogaster.

    PubMed

    Rottschaefer, Susan M; Lazzaro, Brian P

    2012-01-01

    Multiple studies have shown that infection with the endosymbiotic bacterium Wolbachia pipientis confers Drosophila melanogaster and other insects with resistance to infection by RNA viruses. Studies investigating whether Wolbachia infection induces the immune system or confers protection against secondary bacterial infection have not shown any effect. These studies, however, have emphasized resistance against extracellular pathogens. Since Wolbachia lives inside the host cell, we hypothesized that Wolbachia might confer resistance to pathogens that establish infection by invading host cells. We therefore tested whether Wolbachia-infected D. melanogaster are protected against infection by the intracellular pathogenic bacteria Listeria monocytogenes and Salmonella typhimurium, as well as the extracellular pathogenic bacterium Providencia rettgeri. We evaluated the ability of flies infected with Wolbachia to suppress secondary infection by pathogenic bacteria relative to genetically matched controls that had been cured of Wolbachia by treatment with tetracycline. We found no evidence that Wolbachia alters host ability to suppress proliferation of any of the three pathogenic bacteria. Our results indicate that Wolbachia-induced antiviral protection does not result from a generalized response to intracellular pathogens. PMID:22808174

  7. 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

  8. Wolbachia Utilizes Host Microtubules and Dynein for Anterior Localization in the Drosophila Oocyte

    PubMed Central

    Li, Jennifer M; Cao, Jian; Wieschaus, Eric; Sullivan, William

    2005-01-01

    To investigate the role of the host cytoskeleton in the maternal transmission of the endoparasitic bacteria Wolbachia, we have characterized their distribution in the female germ line of Drosophila melanogaster. In the germarium, Wolbachia are distributed to all germ cells of the cyst, establishing an early infection in the cell destined to become the oocyte. During mid-oogenesis, Wolbachia exhibit a distinct concentration between the anterior cortex and the nucleus in the oocyte, where many bacteria appear to contact the nuclear envelope. Following programmed rearrangement of the microtubule network, Wolbachia dissociate from this anterior position and become dispersed throughout the oocyte. This localization pattern is distinct from mitochondria and all known axis determinants. Manipulation of microtubules and cytoplasmic Dynein and Dynactin, but not Kinesin-1, disrupts anterior bacterial localization in the oocyte. In live egg chambers, Wolbachia exhibit movement in nurse cells but not in the oocyte, suggesting that the bacteria are anchored by host factors. In addition, we identify mid-oogenesis as a period in the life cycle of Wolbachia in which bacterial replication occurs. Total bacterial counts show that Wolbachia increase at a significantly higher rate in the oocyte than in the average nurse cell, and that normal Wolbachia levels in the oocyte depend on microtubules. These findings demonstrate that Wolbachia utilize the host microtubule network and associated proteins for their subcellular localization in the Drosophila oocyte. These interactions may also play a role in bacterial motility and replication, ultimately leading to the bacteria's efficient maternal transmission. PMID:16228015

  9. Symbionts Commonly Provide Broad Spectrum Resistance to Viruses in Insects: A Comparative Analysis of Wolbachia Strains

    PubMed Central

    Martinez, Julien; Longdon, Ben; Bauer, Simone; Chan, Yuk-Sang; Miller, Wolfgang J.; Bourtzis, Kostas; Teixeira, Luis; Jiggins, Francis M.

    2014-01-01

    In the last decade, bacterial symbionts have been shown to play an important role in protecting hosts against pathogens. Wolbachia, a widespread symbiont in arthropods, can protect Drosophila and mosquito species against viral infections. We have investigated antiviral protection in 19 Wolbachia strains originating from 16 Drosophila species after transfer into the same genotype of Drosophila simulans. We found that approximately half of the strains protected against two RNA viruses. Given that 40% of terrestrial arthropod species are estimated to harbour Wolbachia, as many as a fifth of all arthropods species may benefit from Wolbachia-mediated protection. The level of protection against two distantly related RNA viruses – DCV and FHV – was strongly genetically correlated, which suggests that there is a single mechanism of protection with broad specificity. Furthermore, Wolbachia is making flies resistant to viruses, as increases in survival can be largely explained by reductions in viral titer. Variation in the level of antiviral protection provided by different Wolbachia strains is strongly genetically correlated to the density of the bacteria strains in host tissues. We found no support for two previously proposed mechanisms of Wolbachia-mediated protection — activation of the immune system and upregulation of the methyltransferase Dnmt2. The large variation in Wolbachia's antiviral properties highlights the need to carefully select Wolbachia strains introduced into mosquito populations to prevent the transmission of arboviruses. PMID:25233341

  10. Diversity and phylogeny of Wolbachia infecting Bactrocera dorsalis (Diptera: Tephritidae) populations from China.

    PubMed

    Sun, Xiao; Cui, Liwang; Li, Zhihong

    2007-10-01

    Wolbachia are a common and widespread group of symbiotic bacteria found in the reproductive tissues of arthropods. Bactrocera dorsalis (Hendel) is an important pest causing considerable economic losses of fruits and vegetables in several southern provinces of China. In this study, polymerase chain reaction (PCR) with general Wolbachia surface protein (wsp) primers was used to test the presence of Wolbachia in 1,500 individuals of B. dorsalis from five geographical populations of China. We detected 19 individuals of B. dorsalis infected by Wolbachia, and the infection rates of different populations varied. Comparison of wsp gene sequences from 19 individuals and search of the GenBank identified four new sequences, probably representing four Wolbachia strains. Sequence comparison showed that the four Wolbachia strains from B. dorsalis in China belonged to three groups (Kue, Mel, and Cuc). Phylogenetic analysis of the wsp sequences suggests that geographical isolation of Wolbachia exists among the populations of B. dorsalis in China, and gene flow of Wolbachia might have occurred between B. dorsalis populations of China and Thailand. Phylogenetic analysis performed on the host mitochondrial cytochrome oxidase I (COI) gene and wsp gene suggests that host has coevolved with Wolbachia. PMID:18284754

  11. 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

  12. 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.

  13. 14 CFR 91.517 - Passenger information.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 14 Aeronautics and Space 2 2010-01-01 2010-01-01 false Passenger information. 91.517 Section 91... Multiengine Airplanes and Fractional Ownership Program Aircraft § 91.517 Passenger information. (a) Except as... belts and when smoking is prohibited. (c) If passenger information signs are installed, no passenger...

  14. 46 CFR 176.113 - Passengers permitted.

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ... 46 Shipping 7 2010-10-01 2010-10-01 false Passengers permitted. 176.113 Section 176.113 Shipping COAST GUARD, DEPARTMENT OF HOMELAND SECURITY (CONTINUED) SMALL PASSENGER VESSELS (UNDER 100 GROSS TONS) INSPECTION AND CERTIFICATION Certificate of Inspection § 176.113 Passengers permitted. (a) The maximum number of passengers permitted must be...

  15. 46 CFR 115.113 - Passengers permitted.

    Code of Federal Regulations, 2012 CFR

    2012-10-01

    ... 46 Shipping 4 2012-10-01 2012-10-01 false Passengers permitted. 115.113 Section 115.113 Shipping COAST GUARD, DEPARTMENT OF HOMELAND SECURITY (CONTINUED) SMALL PASSENGER VESSELS CARRYING MORE THAN 150 PASSENGERS OR WITH OVERNIGHT ACCOMMODATIONS FOR MORE THAN 49 PASSENGERS INSPECTION AND CERTIFICATION General Provisions; Certificate of...

  16. 46 CFR 115.113 - Passengers permitted.

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ... 46 Shipping 4 2013-10-01 2013-10-01 false Passengers permitted. 115.113 Section 115.113 Shipping COAST GUARD, DEPARTMENT OF HOMELAND SECURITY (CONTINUED) SMALL PASSENGER VESSELS CARRYING MORE THAN 150 PASSENGERS OR WITH OVERNIGHT ACCOMMODATIONS FOR MORE THAN 49 PASSENGERS INSPECTION AND CERTIFICATION General Provisions; Certificate of...

  17. 46 CFR 115.113 - Passengers permitted.

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    ... 46 Shipping 4 2014-10-01 2014-10-01 false Passengers permitted. 115.113 Section 115.113 Shipping COAST GUARD, DEPARTMENT OF HOMELAND SECURITY (CONTINUED) SMALL PASSENGER VESSELS CARRYING MORE THAN 150 PASSENGERS OR WITH OVERNIGHT ACCOMMODATIONS FOR MORE THAN 49 PASSENGERS INSPECTION AND CERTIFICATION General Provisions; Certificate of...

  18. 46 CFR 115.113 - Passengers permitted.

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ... 46 Shipping 4 2011-10-01 2011-10-01 false Passengers permitted. 115.113 Section 115.113 Shipping COAST GUARD, DEPARTMENT OF HOMELAND SECURITY (CONTINUED) SMALL PASSENGER VESSELS CARRYING MORE THAN 150 PASSENGERS OR WITH OVERNIGHT ACCOMMODATIONS FOR MORE THAN 49 PASSENGERS INSPECTION AND CERTIFICATION Certificate of Inspection § 115.113...

  19. 19 CFR 4.50 - Passenger lists.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 19 Customs Duties 1 2010-04-01 2010-04-01 false Passenger lists. 4.50 Section 4.50 Customs Duties... VESSELS IN FOREIGN AND DOMESTIC TRADES Passengers on Vessels § 4.50 Passenger lists. (a) The master of... passenger and crew lists, as required by § 4.7(a) of this part. If the vessel is arriving from...

  20. 76 FR 28998 - Implementation of Revised Passenger Weight Standards for Existing Passenger Vessels

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-05-19

    ... FR 78064, December 14, 2010), which updated the AAWPP for new and existing inspected passenger... SECURITY Coast Guard Implementation of Revised Passenger Weight Standards for Existing Passenger Vessels... Passenger Weight Standards for Existing Passenger Vessels.'' This policy letter provides guidance on how...

  1. High Virulence of Wolbachia after Host Switching: When Autophagy Hurts

    PubMed Central

    Le Clec'h, Winka; Braquart-Varnier, Christine; Raimond, Maryline; Ferdy, Jean-Baptiste; Bouchon, Didier; Sicard, Mathieu

    2012-01-01

    Wolbachia are widespread endosymbionts found in a large variety of arthropods. While these bacteria are generally transmitted vertically and exhibit weak virulence in their native hosts, a growing number of studies suggests that horizontal transfers of Wolbachia to new host species also occur frequently in nature. In transfer situations, virulence variations can be predicted since hosts and symbionts are not adapted to each other. Here, we describe a situation where a Wolbachia strain (wVulC) becomes a pathogen when transfected from its native terrestrial isopod host species (Armadillidium vulgare) to another species (Porcellio d. dilatatus). Such transfer of wVulC kills all recipient animals within 75 days. Before death, animals suffer symptoms such as growth slowdown and nervous system disorders. Neither those symptoms nor mortalities were observed after injection of wVulC into its native host A. vulgare. Analyses of wVulC's densities in main organs including Central Nervous System (CNS) of both naturally infected A. vulgare and transfected P. d. dilatatus and A. vulgare individuals revealed a similar pattern of host colonization suggesting an overall similar resistance of both host species towards this bacterium. However, for only P. d. dilatatus, we observed drastic accumulations of autophagic vesicles and vacuoles in the nerve cells and adipocytes of the CNS from individuals infected by wVulC. The symptoms and mortalities could therefore be explained by this huge autophagic response against wVulC in P. d. dilatatus cells that is not triggered in A. vulgare. Our results show that Wolbachia (wVulC) can lead to a pathogenic interaction when transferred horizontally into species that are phylogenetically close to their native hosts. This change in virulence likely results from the autophagic response of the host, strongly altering its tolerance to the symbiont and turning it into a deadly pathogen. PMID:22876183

  2. High virulence of Wolbachia after host switching: when autophagy hurts.

    PubMed

    Le Clec'h, Winka; Braquart-Varnier, Christine; Raimond, Maryline; Ferdy, Jean-Baptiste; Bouchon, Didier; Sicard, Mathieu

    2012-01-01

    Wolbachia are widespread endosymbionts found in a large variety of arthropods. While these bacteria are generally transmitted vertically and exhibit weak virulence in their native hosts, a growing number of studies suggests that horizontal transfers of Wolbachia to new host species also occur frequently in nature. In transfer situations, virulence variations can be predicted since hosts and symbionts are not adapted to each other. Here, we describe a situation where a Wolbachia strain (wVulC) becomes a pathogen when transfected from its native terrestrial isopod host species (Armadillidium vulgare) to another species (Porcellio d. dilatatus). Such transfer of wVulC kills all recipient animals within 75 days. Before death, animals suffer symptoms such as growth slowdown and nervous system disorders. Neither those symptoms nor mortalities were observed after injection of wVulC into its native host A. vulgare. Analyses of wVulC's densities in main organs including Central Nervous System (CNS) of both naturally infected A. vulgare and transfected P. d. dilatatus and A. vulgare individuals revealed a similar pattern of host colonization suggesting an overall similar resistance of both host species towards this bacterium. However, for only P. d. dilatatus, we observed drastic accumulations of autophagic vesicles and vacuoles in the nerve cells and adipocytes of the CNS from individuals infected by wVulC. The symptoms and mortalities could therefore be explained by this huge autophagic response against wVulC in P. d. dilatatus cells that is not triggered in A. vulgare. Our results show that Wolbachia (wVulC) can lead to a pathogenic interaction when transferred horizontally into species that are phylogenetically close to their native hosts. This change in virulence likely results from the autophagic response of the host, strongly altering its tolerance to the symbiont and turning it into a deadly pathogen. PMID:22876183

  3. Wolbachia and its implications for the immunopathology of filariasis.

    PubMed

    Genchi, Claudio; Kramer, Laura H; Sassera, Davide; Bandi, Claudio

    2012-03-01

    Filarial infections are characterized by immunopathological phenomena, that are responsible for the onset of often dramatic pathological outcomes, such as blindness (Onchocerca volvulus) and elephantiasis (W. bancrofti). In addition, the long-term survival (as long as 10 years) of these parasites in otherwise immunocompetent hosts indicates that these nematodes are capable of manipulating the host immune response. The ground-breaking discovery of the bacterial endosymbiont Wolbachia, which resides in most filarial nematodes causing disease, has led to increasing interest in the role it may play in immuno-modulation, pro-inflammatory pathology and other aspects of filarial infection. Indeed, Wolbachia has been shown to be responsible for exacerbating inflammation (as in river blindness), while at the same time blocking efficient elimination of parasites through the host immune response (Onchocerca ochengi). While studies aimed at identifying Wolbachia as a potential target for anti-filarial therapy are at the forefront of current research, understanding its role in the immunology of filarial infection is a fascinating field that has yet to uncover many secrets. PMID:22214329

  4. 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

  5. Facilitating Wolbachia introductions into mosquito populations through insecticide-resistance selection

    PubMed Central

    Hoffmann, Ary A.; Turelli, Michael

    2013-01-01

    Wolbachia infections are being introduced into mosquito vectors of human diseases following the discovery that they can block transmission of disease agents. This requires mosquitoes infected with the disease-blocking Wolbachia to successfully invade populations lacking the infection. While this process is facilitated by features of Wolbachia, particularly their ability to cause cytoplasmic incompatibility, blocking Wolbachia may produce deleterious effects, such as reduced host viability or fecundity, that inhibit successful local introductions and subsequent spatial spread. Here, we outline an approach to facilitate the introduction and spread of Wolbachia infections by coupling Wolbachia introduction to resistance to specific classes of insecticides. The approach takes advantage of very high maternal transmission fidelity of Wolbachia infections in mosquitoes, complete incompatibility between infected males and uninfected females, the widespread occurrence of insecticide resistance, and the widespread use of chemical control in disease-endemic countries. This approach is easily integrated into many existing control strategies, provides population suppression during release and might be used to introduce Wolbachia infections even with high and seasonally dependent deleterious effects, such as the wMelPop infection introduced into Aedes aegypti for dengue control. However, possible benefits will need to be weighed against concerns associated with the introduction of resistance alleles. PMID:23576788

  6. Diversity and phylogenetic relationships of Wolbachia in Drosophila and other native Hawaiian insects

    PubMed Central

    Bennett, Gordon M.; Pantoja, Norma A.; O’Grady, Patrick M.

    2012-01-01

    Wolbachia is a genus of parasitic alphaproteobacteria found in arthropods and nematodes, and represents on of the most common, widespread endosymbionts known. Wolbachia affects a variety of reproductive functions in its host (e.g., male killing, cytoplasmic incompatibility, parthenogenesis), which have the potential to dramatically impact host evolution and species formation. Here, we present the first broad-scale study to screen natural populations of native Hawaiian insects for Wolbachia, focusing on the endemic Diptera. Results indicate that Wolbachia infects native Hawaiian taxa, with alleles spanning phylogenetic supergroups, A and B. The overall frequency of Wolbachia incidene in Hawaiian insects was 14%. The incidence of infection in native Hawaiian Diptera was 11% for individuals and 12% for all species screened. Wolbachia was not detected in two large, widespread Hawaiian dipteran families—Dolichopodidae (44 spp screened) and Limoniidae (12 spp screened). Incidence of infection within endemic Hawaiian lineages that carry Wolbachia was 18% in Drosophilidae species, 25% in Caliphoridae species, > 90% in Nesophrosyne species, 20% in Drosophila dasycnemia and 100% in Nesophrosyne craterigena. Twenty unique alleles were recovered in this study, of which 18 are newly recorded. Screening of endemic populations of D. dasycnemia across Hawaii Island revealed 4 unique alleles. Phylogenetic relationships and allele diversity provide evidence for horizontal transfer of Wolbachia among Hawaiian arthropod lineages. PMID:22878693

  7. 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...

  8. Modeling the indirect effect of Wolbachia on the infection dynamics of horizontally transmitted viruses.

    PubMed

    Strauß, Jakob F; Telschow, Arndt

    2015-01-01

    Intracellular bacteria of the genus Wolbachia are widely distributed in arthropods. There is growing empirical evidence that Wolbachia directly interacts with viruses and other parasites inside the arthropod host, sometimes resulting in low or no pathogen replication. Previous theoretical studies showed that this direct effect of Wolbachia can result in a reduced virus prevalence (within the population), suggesting that Wolbachia could be used in the biological control of vector-borne diseases (e.g., dengue fever). However, Wolbachia might also indirectly affect virus dynamics because Wolbachia-induced reproductive phenotypes (cytoplasmic incompatibility or male killing) increase the larval mortality of hosts and thus alter the age structure of populations. We investigated this indirect effect using mathematical models with overlapping generations, and found the results to depend strongly on the host's life history. In general, the indirect effect can result in two different outcomes: (1) reduced virus prevalence and virus invasion ability, and (2) increased virus prevalence and virus invasion ability. The former occurs for host species with larval competition and undercompensation, the latter for hosts with either adult competition or larval competition and overcompensation. These findings suggest that the effect of Wolbachia on a specific virus is sensitive to the host's life history. We discuss the results with respect to biocontrol programs using Wolbachia. PMID:25972858

  9. Effects on male fitness of removing Wolbachia infections from the mosquito Aedes albopictus.

    PubMed

    Calvitti, M; Moretti, R; Porretta, D; Bellini, R; Urbanelli, S

    2009-06-01

    Cytoplasmic incompatibility (CI) induced by maternally inherited Wolbachia bacteria is a potential tool for the suppression of insect pest species with appropriate patterns of infection. The Asian tiger mosquito Aedes albopictus (Skuse) (Diptera: Culicidae) is known to be infected by two strains of Wolbachia pipientis Hertig (Rickettsiales: Rickettsiaceae), wAlb A and wAlb B, throughout its geographical distribution. This infection pattern theoretically restricts the application of CI-based control strategies. However, Wolbachia can be horizontally transferred using embryonic microinjection to generate incompatible transfected lines harbouring a single new strain of Wolbachia. In order to assess the feasibility of this approach, the effects of Wolbachia removal on mosquito fitness need to be clearly evaluated as the removal of natural superinfection is an inescapable step of this approach. Previous research has shown that uninfected females, produced by antibiotic treatment, showed a decrease in fitness compared with those infected with Wolbachia. In this study, the effect of Wolbachia removal on male fitness was investigated. Longevity and reproductive potential (mating competitiveness and sperm capacity) were assessed in both laboratory cages and greenhouses. No differences were observed between uninfected and infected males with respect to longevity, mating rate, sperm capacity and mating competitiveness in either laboratory conditions or greenhouses. The preservation of fitness in males of Ae. albopictus deprived of natural Wolbachia infection is discussed in relation to the development of incompatible insect technique suppression strategies. Finally, the potential application of aposymbiotic males in mark-release-recapture studies is suggested. PMID:19292821

  10. Cannibalism and Predation as Paths for Horizontal Passage of Wolbachia between Terrestrial Isopods

    PubMed Central

    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. PMID:23593179

  11. 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. PMID:23593179

  12. 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. PMID:24324814

  13. 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

  14. Influence of oxidative homeostasis on bacterial density and cost of infection in Drosophila-Wolbachia symbioses.

    PubMed

    Monnin, D; Kremer, N; Berny, C; Henri, H; Dumet, A; Voituron, Y; Desouhant, E; Vavre, F

    2016-06-01

    The evolution of symbioses along the continuum between parasitism and mutualism can be influenced by the oxidative homeostasis, that is the balance between reactive oxygen species (ROS) and antioxidant molecules. Indeed, ROS can contribute to the host immune defence to regulate symbiont populations, but are also toxic. This interplay between ROS and symbiosis is notably exemplified by recent results in arthropod-Wolbachia interactions. Wolbachia are symbiotic bacteria involved in a wide range of interactions with their arthropods hosts, from facultative, parasitic associations to obligatory, mutualistic ones. In this study, we used Drosophila-Wolbachia associations to determine whether the oxidative homeostasis plays a role in explaining the differences between phenotypically distinct arthropod-Wolbachia symbioses. We used Drosophila lines with different Wolbachia infections and measured the effects of pro-oxidant (paraquat) and antioxidant (glutathione) treatments on the Wolbachia density and the host survival. We show that experimental manipulations of the oxidative homeostasis can reduce the cost of the infection through its effect on Wolbachia density. We discuss the implication of this result from an evolutionary perspective and argue that the oxidative homeostasis could underlie the evolution of tolerance and dependence on Wolbachia. PMID:26999590

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

    PubMed

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

    2016-06-01

    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

  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. A Cell-Based Screen Reveals that the Albendazole Metabolite, Albendazole Sulfone, Targets Wolbachia

    PubMed Central

    Bray, Walter M.; White, Pamela M.; Ruybal, Jordan; Lokey, R. Scott; Debec, Alain; Sullivan, William

    2012-01-01

    Wolbachia endosymbionts carried by filarial nematodes give rise to the neglected diseases African river blindness and lymphatic filariasis afflicting millions worldwide. Here we identify new Wolbachia-disrupting compounds by conducting high-throughput cell-based chemical screens using a Wolbachia-infected, fluorescently labeled Drosophila cell line. This screen yielded several Wolbachia-disrupting compounds including three that resembled Albendazole, a widely used anthelmintic drug that targets nematode microtubules. Follow-up studies demonstrate that a common Albendazole metabolite, Albendazole sulfone, reduces intracellular Wolbachia titer both in Drosophila melanogaster and Brugia malayi, the nematode responsible for lymphatic filariasis. Significantly, Albendazole sulfone does not disrupt Drosophila microtubule organization, suggesting that this compound reduces titer through direct targeting of Wolbachia. Accordingly, both DNA staining and FtsZ immunofluorescence demonstrates that Albendazole sulfone treatment induces Wolbachia elongation, a phenotype indicative of binary fission defects. This suggests that the efficacy of Albendazole in treating filarial nematode-based diseases is attributable to dual targeting of nematode microtubules and their Wolbachia endosymbionts. PMID:23028321

  18. Nonrandom Wolbachia Infection Status of Drosophila melanogaster Strains with Different mtDNA Haplotypes

    PubMed Central

    Nunes, Maria D. S.; Nolte, Viola

    2008-01-01

    Wolbachia are maternally inherited bacteria, which typically spread in the host population by inducing cytoplasmic incompatibility (CI). In Drosophila melanogaster, Wolbachia is quite common but CI is variable, with most of the studies reporting low levels of CI. Surveying mitochondrial DNA (mtDNA) variation and infection status in a worldwide D. melanogaster collection, we found that the Wolbachia infection was not randomly distributed among flies with different mtDNA haplotypes. This preferential infection of some mtDNA haplotypes could be caused by a recent spread of mtDNA haplotypes associated with the infection. The comparison of contemporary D. melanogaster samples with lines collected more than 50 years ago shows that indeed one haplotype with a high incidence of Wolbachia infection has increased in frequency. Consistent with this observation, we found that the acquisition of a Wolbachia infection in a population from Crete was accompanied with an almost complete mtDNA replacement, with the Wolbachia-associated haplotype becoming abundant. Although it is difficult to identify the evolutionary forces causing the global increase of wMel, the parallel sweep of Wolbachia and an mtDNA haplotype suggests a fitness advantage of the Wolbachia infection. PMID:18780877

  19. The endosymbiotic bacterium Wolbachia induces resistance to dengue virus in Aedes aegypti.

    PubMed

    Bian, Guowu; Xu, Yao; Lu, Peng; Xie, Yan; Xi, Zhiyong

    2010-04-01

    Genetic strategies that reduce or block pathogen transmission by mosquitoes have been proposed as a means of augmenting current control measures to reduce the growing burden of vector-borne diseases. The endosymbiotic bacterium Wolbachia has long been promoted as a potential vehicle for introducing disease-resistance genes into mosquitoes, thereby making them refractory to the human pathogens they transmit. Given the large overlap in tissue distribution and intracellular localization between Wolbachia and dengue virus in mosquitoes, we conducted experiments to characterize their interactions. Our results show that Wolbachia inhibits viral replication and dissemination in the main dengue vector, Aedes aegypti. Moreover, the virus transmission potential of Wolbachia-infected Ae. aegypti was significantly diminished when compared to wild-type mosquitoes that did not harbor Wolbachia. At 14 days post-infection, Wolbachia completely blocked dengue transmission in at least 37.5% of Ae. aegypti mosquitoes. We also observed that this Wolbachia-mediated viral interference was associated with an elevated basal immunity and increased longevity in the mosquitoes. These results underscore the potential usefulness of Wolbachia-based control strategies for population replacement. PMID:20368968

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

    PubMed

    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

  1. Wolbachia Induces Male-Specific Mortality in the Mosquito Culex pipiens (LIN Strain)

    PubMed Central

    Rasgon, Jason L.

    2012-01-01

    Background Wolbachia are maternally inherited endosymbionts that infect a diverse range of invertebrates, including insects, arachnids, crustaceans and filarial nematodes. Wolbachia are responsible for causing diverse reproductive alterations in their invertebrate hosts that maximize their transmission to the next generation. Evolutionary theory suggests that due to maternal inheritance, Wolbachia should evolve toward mutualism in infected females, but strict maternal inheritance means there is no corresponding force to select for Wolbachia strains that are mutualistic in males. Methodology/Principal findings Using cohort life-table analysis, we demonstrate that in the mosquito Culex pipiens (LIN strain), Wolbachia-infected females show no fitness costs due to infection. However, Wolbachia induces up to a 30% reduction in male lifespan. Conclusions/significance These results indicate that the Wolbachia infection of the Culex pipiens LIN strain is virulent in a sex-specific manner. Under laboratory situations where mosquitoes generally mate at young ages, Wolbachia strains that reduce male survival could evolve by drift because increased mortality in older males is not a significant selective force. PMID:22427798

  2. 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...

  3. 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. PMID:26070017

  4. Characterization and transcriptional analysis of two gene clusters for type IV secretion machinery in Wolbachia of Armadillidium vulgare.

    PubMed

    Félix, Christine; Pichon, Samuel; Braquart-Varnier, Christine; Braig, Henk; Chen, Lanming; Garrett, Roger A; Martin, Gilbert; Grève, Pierre

    2008-01-01

    Wolbachia are maternally inherited alpha-proteobacteria that induce feminization of genetic males in most terrestrial crustacean isopods. Two clusters of vir genes for a type IV secretion machinery have been identified at two separate loci and characterized for the first time in a feminizing Wolbachia. Furthermore, we demonstrated that these operons are transcriptionally active in ovaries and in all other tissues tested, suggesting that T4SS has a significant role in Wolbachia biology. These observations and the identification of homologous vir genes in Wolbachia strains infecting insects or nematodes show that vir genes are conserved among Wolbachia strains whatever the phenotype induced by the bacteria. PMID:18582562

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

    PubMed

    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. 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

  7. Effect of Wolbachia on insecticide susceptibility in lines of Aedes aegypti.

    PubMed

    Endersby, N M; Hoffmann, A A

    2013-06-01

    Two stable infections of Wolbachia pipientis, wMelPop and wMel, now established in Aedes aegypti, are being used in a biocontrol program to suppress the transmission of dengue. Any effects of Wolbachia infection on insecticide resistance of mosquitoes may undermine the success of this program. Bioassays of Ae. aegypti were conducted to test for differences in response to insecticides between Wolbachia infected (wMelPop, wMel) and uninfected lines. Insecticides screened were bifenthrin, the pyrethroid commonly used for adult knockdown, as well as larvicides: Bacillus thuringiensis var. israelensis, the organophosphate, temephos and the insect growth regulator, s-methoprene. While differences in response between lines were detected for some insecticides, no obvious or consistent effects related to presence of Wolbachia infection were observed. Spreading Wolbachia infections are, therefore, unlikely to affect the efficacy of traditional chemical control of mosquito outbreaks. PMID:23149015

  8. DETECTION OF THE WOLBACHIA-ENCODED DNA BINDING PROTEIN, HU beta, IN MOSQUITO GONADS

    PubMed Central

    BECKMANN, JOHN F.; MARKOWSKI, TODD W.; WITTHUHN, BRUCE A.; FALLON, ANN M.

    2013-01-01

    Wolbachia are obligate intracellular bacteria that cause cytoplasmic incompatibility in mosquitoes. In an incompatible cross, eggs of uninfected females fail to hatch when fertilized by sperm from infected males. We used polyacrylamide gel electrophoresis and tandem mass spectrometry to identify Wolbachia proteins in infected mosquito gonads. These included surface proteins with masses of 25 and 18 kDa and the DNA binding protein, HU beta. Using reverse transcriptase polymerase chain reaction, we showed that the HU gene is transcribed in Wolbachia-infected Culex pipiens and Aedes albopictus mosquitoes. We sequenced HU genes from four Wolbachia strains and compared deduced protein sequences with additional homologs from the databases. Among the Rickettsiales, Wolbachia HU has distinct N- and C-terminal basic/acidic amino acid motifs as well as a pair of conserved, cysteine residues. PMID:23287400

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

    PubMed

    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

  10. 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

  11. 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

  12. 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

  13. 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. PMID:24750488

  14. 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. PMID:27078260

  15. Transinfected Wolbachia have minimal effects on male reproductive success in Aedes aegypti

    PubMed Central

    2013-01-01

    Background Wolbachia are maternally inherited endosymbiotic bacteria that manipulate the reproductive success of their insect hosts. Uninfected females that mate with Wolbachia infected males do not reproduce due to cytoplasmic incompatibility (CI). CI results in the increased frequency of Wolbachia-infected individuals in populations. Recently, two Wolbachia strains, the benign wMel and virulent wMelPop have been artificially transinfected into the primary vector of dengue virus, the mosquito Ae. aegypti where they have formed stable infections. These Wolbachia infections are being developed for a biological control strategy against dengue virus transmission. While the effects of Wolbachia on female Ae. aegypti have been examined the effects on males are less well characterised. Here we ascertain and compare the effects of the two strains on male fitness in resource-limited environments that may better approximate the natural environment. Methods A series of population mating trials were conducted to examine the effect of Wolbachia infection status (with strains wMel and wMelPop) and male larval nutrition on insemination frequency, remating rates, the fecundity of females, the hatch rates of eggs and the wing length and fertility of males. Results wMel and wMelPop infections reduce the fecundity of infected females and wMelPop reduces the viability of eggs. Low nutrition diets for males in the larval phase affects the fecundity of wMel-infected females. Neither strain of Wolbachia affected sperm quality or viability or the ability of males to successfully mate multiple females. Conclusions The benign strain of Wolbachia, wMel causes similar reductions in fecundity as the more virulent, wMelPop, and neither are too great that they should not still spread given the action of CI. The ability of Wolbachia-infected males to repeat mate as frequently as wildtype mosquitoes indicates that they will be very good agents of delivering CI in field release populations. PMID

  16. Discovery of a new Wolbachia supergroup in cave spider species and the lateral transfer of phage WO among distant hosts.

    PubMed

    Wang, Guan-Hong; Jia, Ling-Yi; Xiao, Jin-Hua; Huang, Da-Wei

    2016-07-01

    Wolbachia are widespread intracellular bacteria infecting the major classes of arthropods and some filarial nematodes. In arthropods, Wolbachia have evolved various intriguing reproductive manipulations, including cytoplasmic incompatibility, parthenogenesis, feminization, and male killing. Sixteen supergroups of Wolbachia have been identified, named A-Q (except G). Though Wolbachia present great diversity in arthropods, spiders, especially cave spiders, are still a poorly surveyed group of Wolbachia hosts. Here, we report a novel Wolbachia supergroup from nine Telema cave spiders (Araneae: Telemidae) based on five molecular markers (16S rRNA, ftsZ, gltA, groEL, and coxA). In addition, phage WO, which was previously reported only in Wolbachia supergroups A, B, and F, infects this new Wolbachia supergroup. We detected a 100% infection rate for phage WO and Wolbachia in Telema species. The phylogenetic trees of phage WO and Wolbachia are not congruent, which suggests that horizontal transfer of phage WO has occurred in these secluded species. Additionally, these data indicate Telema-Wolbachia-phage WO may be a good model for exploring the horizontal transfer history of WO among different host species. PMID:26997548

  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. PMID:25812130

  18. Wolbachia in the flesh: symbiont intensities in germ-line and somatic tissues challenge the conventional view of Wolbachia transmission routes.

    PubMed

    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

  19. 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

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

    PubMed Central

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

    2009-01-01

    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. PMID:19364731

  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. 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. PMID:26631376

  3. 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

  4. Structural and functional characterization of the oxidoreductase alpha-DsbA1 from Wolbachia pipientis.

    PubMed

    Kurz, Mareike; Iturbe-Ormaetxe, Iñaki; Jarrott, Russell; Shouldice, Stephen R; Wouters, Merridee A; Frei, Patrick; Glockshuber, Rudi; O'Neill, Scott L; Heras, Begoña; Martin, Jennifer L

    2009-07-01

    The alpha-proteobacterium Wolbachia pipientis is a highly successful intracellular endosymbiont of invertebrates that manipulates its host's reproductive biology to facilitate its own maternal transmission. The fastidious nature of Wolbachia and the lack of genetic transformation have hampered analysis of the molecular basis of these manipulations. Structure determination of key Wolbachia proteins will enable the development of inhibitors for chemical genetics studies. Wolbachia encodes a homologue (alpha-DsbA1) of the Escherichia coli dithiol oxidase enzyme EcDsbA, essential for the oxidative folding of many exported proteins. We found that the active-site cysteine pair of Wolbachia alpha-DsbA1 has the most reducing redox potential of any characterized DsbA. In addition, Wolbachia alpha-DsbA1 possesses a second disulfide that is highly conserved in alpha-proteobacterial DsbAs but not in other DsbAs. The alpha-DsbA1 structure lacks the characteristic hydrophobic features of EcDsbA, and the protein neither complements EcDsbA deletion mutants in E. coli nor interacts with EcDsbB, the redox partner of EcDsbA. The surface characteristics and redox profile of alpha-DsbA1 indicate that it probably plays a specialized oxidative folding role with a narrow substrate specificity. This first report of a Wolbachia protein structure provides the basis for future chemical genetics studies. PMID:19265485

  5. Infection with Wolbachia protects mosquitoes against Plasmodium-induced mortality in a natural system.

    PubMed

    Zélé, F; Nicot, A; Duron, O; Rivero, A

    2012-07-01

    In recent years, there has been a shift in the one host-one parasite paradigm with the realization that, in the field, most hosts are coinfected with multiple parasites. Coinfections are particularly relevant when the host is a vector of diseases, because multiple infections can have drastic consequences for parasite transmission at both the ecological and evolutionary timescales. Wolbachia pipientis is the most common parasitic microorganism in insects, and as such, it is of special interest for understanding the role of coinfections in the outcome of parasite infections. Here, we investigate whether Wolbachia can modulate the effect of Plasmodium on what is, arguably, the most important component of the vectorial capacity of mosquitoes: their longevity. For this purpose, and in contrast to recent studies that have focused on mosquito-Plasmodium and/or mosquito-Wolbachia combinations not found in nature, we work on a Wolbachia-mosquito-Plasmodium triad with a common evolutionary history. Our results show that Wolbachia protects mosquitoes from Plasmodium-induced mortality. The results are consistent across two different strains of Wolbachia and repeatable across two different experimental blocks. To our knowledge, this is the first time that such an effect has been shown for Plasmodium-infected mosquitoes and, in particular, in a natural Wolbachia-host combination. We discuss different mechanistic and evolutionary explanations for these results as well as their consequences for Plasmodium transmission. PMID:22533729

  6. 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. PMID:15138452

  7. Wolbachia endosymbiont responsible for cytoplasmic incompatibility in a terrestrial crustacean: effects in natural and foreign hosts.

    PubMed

    Moret, Y; Juchault, P; Rigaud, T

    2001-03-01

    Wolbachia bacteria are vertically transmitted endosymbionts that disturb the reproduction of many arthropods thereby enhancing their spread in host populations. Wolbachia are often responsible for changes of sex ratios in terrestrial isopods, a result of the feminization of genotypic males. Here we found that the Wolbachia hosted by Cylisticus convexus (wCc) caused unidirectional cytoplasmic incompatibility (CI), an effect commonly found in insects. To understand the diversity of Wolbachia-induced effects in isopods, wCc were experimentally transferred in a novel isopod host, Armadillidium vulgare. wCc conserved the ability to induce CI. However, Wolbachia were not transmitted to the eggs, so the capacity to restore the compatibility in crosses involving two transinfected individuals was lost. The feminizing Wolbachia hosted by A. vulgare was unable to rescue CI induced by wCc. These results showed that Wolbachia in isopods did not evolved broadly to induce feminization, and that CI and the feminizing effect are probably due to different mechanisms. In addition, wCc reduces the mating capacity of infected C. convexus males, suggesting that the bacteria might alter reproductive behaviour. The maintenance of wCc in host populations is discussed. PMID:11488969

  8. 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

    2009-11-01

    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

  9. 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

  10. 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

  11. 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. PMID:27492563

  12. 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

  13. Physiological cost induced by the maternally-transmitted endosymbiont Wolbachia in the Drosophila parasitoid Leptopilina heterotoma.

    PubMed

    Fleury, F; Vavre, F; Ris, N; Fouillet, P; Boulétreau, M

    2000-11-01

    Endosymbiotic bacteria of the genus Wolbachia infect a number of invertebrate species in which they induce various alterations in host reproduction, mainly cytoplasmic incompatibility (CI). In contrast to most other maternally transmitted parasites, manipulation of host reproduction makes the spread of Wolbachia possible even if they induce a physiological cost on their hosts. Current studies have shown that fitness consequences of Wolbachia infection could range from positive (mutualist) to negative (parasitic) but, in most cases, Wolbachia do not have strong deleterious effects on host fitness and the status of association remains unclear. Here, we show that in the Drosophila parasitoid wasp Leptopilina heterotoma, Wolbachia infection has a negative impact on several host fitness traits of both sexes. Fecundity, adult survival and locomotor performance are significantly reduced, whereas circadian rhythm, development time and offspring sex-ratio are not affected. Although the cost of bacterial infection can be overcome by effects on host reproduction i.e. cytoplasmic incompatibility, it could influence the spread of the bacterium at the early stages of the invasion process. Clearly, results underline the wide spectrum of phenotypic effects of Wolbachia infection and, to our knowledge, Wolbachia infection of L. heterotoma appears to be one of the most virulent that has ever been observed in insects. PMID:11128800

  14. Onchocerciasis: the Role of Wolbachia Bacterial Endosymbionts in Parasite Biology, Disease Pathogenesis, and Treatment

    PubMed Central

    Tamarozzi, Francesca; Halliday, Alice; Gentil, Katrin; Hoerauf, Achim; Pearlman, Eric; Taylor, Mark J.

    2011-01-01

    Summary: The discovery of Wolbachia intracellular bacteria within filarial nematodes, including Onchocerca volvulus, the causative agent of onchocerciasis or “river blindness,” has delivered a paradigm shift in our understanding of the parasite's biology, to where we now know that the bacterial endosymbionts are essential for normal development of larvae and embryos and may support the long-term survival of adult worms. The apparent mutualistic dependency has also offered a novel approach to the treatment of onchocerciasis through the use of antibiotics to eliminate Wolbachia, delivering for the first time a treatment which has significant macrofilaricidal efficacy. Studies with other filarial nematode species have also highlighted a role for Wolbachia in transmission and infection of the mammalian host through a fascinating manipulation of mast cell-mediated vasodilation to enhance infectivity of vector-borne larvae. Wolbachia has also been identified as the principal driver of innate and adaptive Th1 inflammatory immunity, which can either contribute to disease pathogenesis or, with the Wolbachia-mediated recruitment of mast cells, enhance infectivity. The Wolbachia activation of innate inflammation also drives inflammatory adverse events in response to chemotherapy with either diethylcarbamazine (DEC) or ivermectin. In this review we summarize the experimental and field trial data which have uncovered the importance of Wolbachia symbiosis in onchocerciasis. PMID:21734243

  15. 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. PMID:27459519

  16. 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

  17. 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

  18. 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

  19. 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. PMID:26531251

  20. Wolbachia Utilize Host Actin for Efficient Maternal Transmission in Drosophila melanogaster

    PubMed Central

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

    2015-01-01

    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 (chic221/+ and chic1320/+) or villin (qua6-396/+) 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 chic221 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. PMID:25906062

  1. Integrated transcriptomic and proteomic analysis of the global response of Wolbachia to doxycycline-induced stress

    PubMed Central

    Darby, Alistair C; Christina Gill, A; Armstrong, Stuart D; Hartley, Catherine S; Xia, Dong; Wastling, Jonathan M; Makepeace, Benjamin L

    2014-01-01

    The bacterium Wolbachia (order Rickettsiales), representing perhaps the most abundant vertically transmitted microbe worldwide, infects arthropods and filarial nematodes. In arthropods, Wolbachia can induce reproductive alterations and interfere with the transmission of several arthropod-borne pathogens. In addition, Wolbachia is an obligate mutualist of the filarial parasites that cause lymphatic filariasis and onchocerciasis in the tropics. Targeting Wolbachia with tetracycline antibiotics leads to sterilisation and ultimately death of adult filariae. However, several weeks of treatment are required, restricting the implementation of this control strategy. To date, the response of Wolbachia to stress has not been investigated, and almost nothing is known about global regulation of gene expression in this organism. We exposed an arthropod Wolbachia strain to doxycycline in vitro, and analysed differential expression by directional RNA-seq and label-free, quantitative proteomics. We found that Wolbachia responded not only by modulating expression of the translation machinery, but also by upregulating nucleotide synthesis and energy metabolism, while downregulating outer membrane proteins. Moreover, Wolbachia increased the expression of a key component of the twin-arginine translocase (tatA) and a phosphate ABC transporter ATPase (PstB); the latter is associated with decreased susceptibility to antimicrobials in free-living bacteria. Finally, the downregulation of 6S RNA during translational inhibition suggests that this small RNA is involved in growth rate control. Despite its highly reduced genome, Wolbachia shows a surprising ability to regulate gene expression during exposure to a potent stressor. Our findings have general relevance for the chemotherapy of obligate intracellular bacteria and the mechanistic basis of persistence in the Rickettsiales. PMID:24152719

  2. 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

  3. 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

  4. Extreme Divergence of Wolbachia Tropism for the Stem-Cell-Niche in the Drosophila Testis

    PubMed Central

    Toomey, Michelle E.; Frydman, Horacio M.

    2014-01-01

    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

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

    PubMed Central

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

    2016-01-01

    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. PMID:26531251

  6. 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

  7. 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

  8. High Levels of Multiple Infections, Recombination and Horizontal Transmission of Wolbachia in the Andricus mukaigawae (Hymenoptera; Cynipidae) Communities

    PubMed Central

    Yang, Xiao-Hui; Zhu, Dao-Hong; Liu, Zhiwei; Zhao, Ling; Su, Cheng-Yuan

    2013-01-01

    Wolbachia are maternally inherited endosymbiotic bacteria of arthropods and nematodes. In arthropods, they manipulate the reproduction of their hosts to facilitate their own spread in host populations, causing cytoplasmic incompatibility, parthenogenesis induction, feminization of genetic males and male-killing. In this study, we investigated Wolbachia infection and studied wsp (Wolbachia surface protein) sequences in three wasp species associated with the unisexual galls of A. mukaigawae with the aim of determining the transmission mode and the reason for multiple infections of Wolbachia. Frequency of Wolbachia infected populations for A. mukaigawae, Synergus japonicus (inquiline), and Torymus sp. (parasitoid) was 75%, 100%, and 100%, respectively. Multiple Wolbachia infections were detected in A. mukaigawae and S. japonicus, with 5 and 8 Wolbachia strains, respectively. The two host species shared 5 Wolbachia strains and were infected by identical strains in several locations, indicating horizontal transmission of Wolbachia. The transmission potentially takes place through gall tissues, which the larvae of both wasps feed on. Furthermore, three recombination events of Wolbachia were observed: the strains W8, W2 and W6 apparently have derived from W3 and W5a, W6 and W7, W4 and W9, respectively. W8 and W2 and their respective parental strains were detected in S. japonicus. W6 was detected with only one parent (W4) in S. japonicus; W9 was detected in Torymus sp., suggesting horizontal transmission between hosts and parasitoids. In conclusion, our research supports earlier studies that horizontal transmission of Wolbachia, a symbiont of the Rickettsiales order, may be plant-mediated or take place between hosts and parasitoids. Our research provides novel molecular evidence for multiple recombination events of Wolbachia in gall wasp communities. We suggest that genomic recombination and potential plant-mediated horizontal transmission may be attributable to the high

  9. Identifying influential nodes in complex networks based on expansion factor

    NASA Astrophysics Data System (ADS)

    Liu, Dong; Jing, Yun; Chang, Baofang

    2016-03-01

    Identifying the top influential spreaders in a network has practical significance. In this paper, we propose a novel centrality to identify influential spreaders based on expansion factor. Nodes with high expansion factor centrality (EFC) have strong spreading capability. During the course of the work, an improved strategy is proposed to reduce the time complexity of EFC. We discuss the correlations between EFC and the other five classical indicators. Simulation results on the Susceptible-Infected-Removed (SIR) model manifest that EFC can identify influential nodes and find some critical influential nodes neglected by other indicators.

  10. 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)...

  11. 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)...

  12. 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)...

  13. 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)...

  14. 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)...

  15. 46 CFR 176.113 - Passengers permitted.

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ... 46 Shipping 7 2013-10-01 2013-10-01 false Passengers permitted. 176.113 Section 176.113 Shipping COAST GUARD, DEPARTMENT OF HOMELAND SECURITY (CONTINUED) SMALL PASSENGER VESSELS (UNDER 100 GROSS TONS) INSPECTION AND CERTIFICATION General Provisions; Certificate of Inspection § 176.113 Passengers permitted. (a) The maximum number of...

  16. 46 CFR 176.113 - Passengers permitted.

    Code of Federal Regulations, 2012 CFR

    2012-10-01

    ... 46 Shipping 7 2012-10-01 2012-10-01 false Passengers permitted. 176.113 Section 176.113 Shipping COAST GUARD, DEPARTMENT OF HOMELAND SECURITY (CONTINUED) SMALL PASSENGER VESSELS (UNDER 100 GROSS TONS) INSPECTION AND CERTIFICATION General Provisions; Certificate of Inspection § 176.113 Passengers permitted. (a) The maximum number of...

  17. 46 CFR 176.113 - Passengers permitted.

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    ... 46 Shipping 7 2014-10-01 2014-10-01 false Passengers permitted. 176.113 Section 176.113 Shipping COAST GUARD, DEPARTMENT OF HOMELAND SECURITY (CONTINUED) SMALL PASSENGER VESSELS (UNDER 100 GROSS TONS) INSPECTION AND CERTIFICATION General Provisions; Certificate of Inspection § 176.113 Passengers permitted. (a) The maximum number of...

  18. 14 CFR 91.517 - Passenger information.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... 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 or crewmember may smoke while any “no smoking” sign is lighted nor may any passenger or crewmember smoke in...

  19. 14 CFR 91.519 - Passenger briefing.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... briefed on— (1) Smoking. Each passenger shall be briefed on when, where, and under what conditions smoking... 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...

  20. 46 CFR 176.113 - Passengers permitted.

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ... permitted for each 760 millimeters (30 inches) of rail space available to the passengers at the periphery of each deck. The following rail space may not be used in determining the maximum number of passengers permitted: (i) Rail space in congested areas unsafe for passengers, such as near anchor handling...

  1. 77 FR 38248 - Passenger Train Emergency Preparedness

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-06-27

    ... rule on passenger train emergency preparedness that was codified at 49 CFR part 239. See 63 FR 24629... evacuate passengers. See 73 FR 6369 (February 1, 2008). While this final rule did not make any changes to... existing requirements as well as create new requirements for passenger train emergency systems. See 77...

  2. 14 CFR 91.517 - Passenger information.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... 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 or crewmember may smoke while any “no smoking” sign is lighted nor may any passenger or crewmember smoke in...

  3. 14 CFR 91.517 - Passenger information.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... 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 or crewmember may smoke while any “no smoking” sign is lighted nor may any passenger or crewmember smoke in...

  4. 14 CFR 91.519 - Passenger briefing.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... briefed on— (1) Smoking. Each passenger shall be briefed on when, where, and under what conditions smoking... 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...

  5. 14 CFR 91.519 - Passenger briefing.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... briefed on— (1) Smoking. Each passenger shall be briefed on when, where, and under what conditions smoking... 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...

  6. 14 CFR 91.517 - Passenger information.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... 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 or crewmember may smoke while any “no smoking” sign is lighted nor may any passenger or crewmember smoke in...

  7. 14 CFR 91.519 - Passenger briefing.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... briefed on— (1) Smoking. Each passenger shall be briefed on when, where, and under what conditions smoking... 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...

  8. PASSENGER CAR HYDROCARBON EMISSIONS SPECIATION

    EPA Science Inventory

    Emission factors for over 60 individual hydrocarbon compounds were determined for four passenger cars. The cars included a 1963 Chevrolet, a 1977 Mustang, and 1978 Monarch, and 1979 LTD II. The speciation data is reported for both tailpipe and evaporative emissions. The tailpipe ...

  9. 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. PMID:26664808

  10. 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

  11. 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…

  12. 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.

  13. Wolbachia Blocks Currently Circulating Zika Virus Isolates in Brazilian Aedes aegypti Mosquitoes.

    PubMed

    Dutra, Heverton Leandro Carneiro; Rocha, Marcele Neves; Dias, Fernando Braga Stehling; Mansur, Simone Brutman; Caragata, Eric Pearce; Moreira, Luciano Andrade

    2016-06-01

    The recent association of Zika virus with cases of microcephaly has sparked a global health crisis and highlighted the need for mechanisms to combat the Zika vector, Aedes aegypti mosquitoes. Wolbachia pipientis, a bacterial endosymbiont of insect, has recently garnered attention as a mechanism for arbovirus control. Here we report that Aedes aegypti harboring Wolbachia are highly resistant to infection with two currently circulating Zika virus isolates from the recent Brazilian epidemic. Wolbachia-harboring mosquitoes displayed lower viral prevalence and intensity and decreased disseminated infection and, critically, did not carry infectious virus in the saliva, suggesting that viral transmission was blocked. Our data indicate that the use of Wolbachia-harboring mosquitoes could represent an effective mechanism to reduce Zika virus transmission and should be included as part of Zika control strategies. PMID:27156023

  14. 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...

  15. Association of a New Wolbachia Strain with, and Its Effects on, Leptopilina victoriae, a Virulent Wasp Parasitic to Drosophila spp.

    PubMed Central

    Gueguen, Gwenaelle; Onemola, Bodunde

    2012-01-01

    Wolbachia bacteria are ubiquitous intracellular bacteria of arthropods. Often considered reproductive parasites, they can benefit certain host species. We describe a new Wolbachia strain from Leptopilina victoriae, a Drosophila wasp. The strain is closely related to Wolbachia from Culex sp. Located to the posterior poles of oocytes, it manipulates its host's reproduction by inducing a male development type of cytoplasmic incompatibility. We also report its diverse effects on the wasp's life history traits. PMID:22685158

  16. 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

  17. 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

  18. Temperature Affects the Tripartite Interactions between Bacteriophage WO, Wolbachia, and Cytoplasmic Incompatibility

    PubMed Central

    Bordenstein, Sarah R.; Bordenstein, Seth R.

    2011-01-01

    Wolbachia infections are a model for understanding intracellular, bacterial symbioses. While the symbiosis is often studied from a binary perspective of host and bacteria, it is increasingly apparent that additional trophic levels can influence the symbiosis. For example, Wolbachia in arthropods harbor a widespread temperate bacteriophage, termed WO, that forms virions and rampantly transfers between coinfections. Here we test the hypothesis that temperatures at the extreme edges of an insect's habitable range alter bacteriophage WO inducibility and in turn, Wolbachia densities and the penetrance of cytoplasmic incompatibility. We report four key findings using the model wasp, Nasonia vitripennis: First, both cold treatment at 18 C and heat treatment at 30 C reduce Wolbachia densities by as much as 74% relative to wasps reared at 25 C. Second, in all cases where Wolbachia densities decline due to temperature changes, phage WO densities increase and inversely associate with Wolbachia densities. Heat has a marked effect on phage WO, yielding phage densities that are 552% higher than the room temperature control. Third, there is a significant affect of insect family on phage WO and endoysmbiont densities. Fourth, at extreme temperatures, there was a temperature-mediated adjustment to the density threshold at which Wolbachia cause complete cytoplasmic incompatibility. Taken together, these results demonstrate that temperature simultaneously affects phage WO densities, endosymbiont densities, and the penetrance of cytoplasmic incompatibility. While temperature shock enhances bacteriophage inducibility and the ensuing bacterial mortality in a wide range of medically and industrially-important bacteria, this is the first investigation of the associations in an obligate intracellular bacteria. Implications to a SOS global sensing feedback mechanism in Wolbachia are discussed. PMID:22194999

  19. The Relative Importance of Innate Immune Priming in Wolbachia-Mediated Dengue Interference

    PubMed Central

    Rancès, Edwige; Ye, Yixin H.; Woolfit, Megan; McGraw, Elizabeth A.; O'Neill, Scott L.

    2012-01-01

    The non-virulent Wolbachia strain wMel and the life-shortening strain wMelPop-CLA, both originally from Drosophila melanogaster, have been stably introduced into the mosquito vector of dengue fever, Aedes aegypti. Each of these Wolbachia strains interferes with viral pathogenicity and/or dissemination in both their natural Drosophila host and in their new mosquito host, and it has been suggested that this virus interference may be due to host immune priming by Wolbachia. In order to identify aspects of the mosquito immune response that might underpin virus interference, we used whole-genome microarrays to analyse the transcriptional response of A. aegypti to the wMel and wMelPop-CLA Wolbachia strains. While wMel affected the transcription of far fewer host genes than wMelPop-CLA, both strains activated the expression of some immune genes including anti-microbial peptides, Toll pathway genes and genes involved in melanization. Because the induction of these immune genes might be associated with the very recent introduction of Wolbachia into the mosquito, we also examined the same Wolbachia strains in their original host D. melanogaster. First we demonstrated that when dengue viruses were injected into D. melanogaster, virus accumulation was significantly reduced in the presence of Wolbachia, just as in A. aegypti. Second, when we carried out transcriptional analyses of the same immune genes up-regulated in the new heterologous mosquito host in response to Wolbachia we found no over-expression of these genes in D. melanogaster, infected with either wMel or wMelPop. These results reinforce the idea that the fundamental mechanism involved in viral interference in Drosophila and Aedes is not dependent on the up-regulation of the immune effectors examined, although it cannot be excluded that immune priming in the heterologous mosquito host might enhance the virus interference trait. PMID:22383881

  20. Human Probing Behavior of Aedes aegypti when Infected with a Life-Shortening Strain of Wolbachia

    PubMed Central

    Moreira, Luciano A.; Saig, Emad; Turley, Andrew P.; Ribeiro, José M. C.; O'Neill, Scott L.; McGraw, Elizabeth A.

    2009-01-01

    Background Mosquitoes are vectors of many serious pathogens in tropical and sub-tropical countries. Current control strategies almost entirely rely upon insecticides, which increasingly face the problems of high cost, increasing mosquito resistance and negative effects on non-target organisms. Alternative strategies include the proposed use of inherited life-shortening agents, such as the Wolbachia bacterium. By shortening mosquito vector lifespan, Wolbachia could potentially reduce the vectorial capacity of mosquito populations. We have recently been able to stably transinfect Aedes aegypti mosquitoes with the life-shortening Wolbachia strain wMelPop, and are assessing various aspects of its interaction with the mosquito host to determine its likely impact on pathogen transmission as well as its potential ability to invade A. aegypti populations. Methodology/Principal Findings Here we have examined the probing behavior of Wolbachia-infected mosquitoes in an attempt to understand both the broader impact of Wolbachia infection on mosquito biology and, in particular, vectorial capacity. The probing behavior of wMelPop-infected mosquitoes at four adult ages was examined and compared to uninfected controls during video-recorded feeding trials on a human hand. Wolbachia-positive insects, from 15 days of age, showed a drastic increase in the time spent pre-probing and probing relative to uninfected controls. Two other important features for blood feeding, saliva volume and apyrase content of saliva, were also studied. Conclusions/Significance As A. aegypti infected with wMelPop age, they show increasing difficulty in completing the process of blood feeding effectively and efficiently. Wolbachia-infected mosquitoes on average produced smaller volumes of saliva that still contained the same amount of apyrase activity as uninfected mosquitoes. These effects on blood feeding behavior may reduce vectorial capacity and point to underlying physiological changes in Wolbachia

  1. Wolbachia from Drosophila incompta: just a hitchhiker shared by Drosophila in the New and Old World?

    PubMed

    Wallau, G L; da Rosa, M T; De Ré, F C; Loreto, E L S

    2016-08-01

    Wolbachia are intracellular endosymbionts that infect arthropods and filarial nematodes, occasionally causing a wide variety of modifications in host biology, such as male-killing and cytoplasmic incompatibility (CI), amongst others. This study assembled draft genomes for Wolbachia infecting Drosophila incompta, a species that uses flowers as exclusive breeding and feeding sites, in two distinct Brazilian populations. The absence of four genes involved in CI from this genome, together with literature reports of low frequencies of infected flies in wild populations that contain high mitogenome polymorphism, suggests that this bacterium does not induce CI in D. incompta. Phylogenomic analysis placed Wolbachia infecting D. incompta as closely related to the wMel strain which received such name since it was originally detected in Drosophila melanogaster. In addition, phylogenetic analysis using the Wolbachia surface protein gene and five genes used for multilocus sequence typing of Wolbachia found infecting Drosophila and other arthropod species of Old and New World displayed a complex evolutionary scenario involving recent horizontal transfer bursts in all major clades of Wolbachia pipens belonging to the supergroup A in both geographical regions. PMID:27122079

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

    PubMed

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

    2014-11-01

    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 desulphurase, 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

  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. The Intracellular Bacterium Wolbachia Uses Parasitoid Wasps as Phoretic Vectors for Efficient Horizontal Transmission

    PubMed Central

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

    2015-01-01

    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. PMID:25675099

  6. Evidence for recombination between feminizing Wolbachia in the isopod genus Armadillidium.

    PubMed

    Verne, Sébastien; Johnson, Monique; Bouchon, Didier; Grandjean, Frédéric

    2007-08-01

    Wolbachia are maternally inherited endosymbiotic alpha-Proteobacteria infecting a wide range of arthropods. Wolbachia induce feminization in many terrestrial isopod species, particularly in the genus Armadillidium (Crustacea, Oniscidea). The diversity of Wolbachia strains infecting Armadillidium species was examined. Results reveal that natural populations of A. vulgare contain three different Wolbachia strains (wVulC, wVulM and wVulP). The wsp gene and its 3'-adjacent region show evidence that two recombination events have occurred between two of these strains. In both cases, multiple statistical analyses suggest that a small gene fragment of a strain closely related to wVulM (minor parent) is inserted into the genome of another strain closely related to wVulC (major parent). Although multiple infections in a single individual have never been demonstrated in natural population, the existence of recombination between feminizing strains suggests that bi-infections are possible, or at least that bi-infections can be maintained sufficiently long enough to allow recombination. Recombination events increase genetic diversity of Wolbachia found in Armadillidium species and may play a role in the ability of Wolbachia strains to invade new hosts. PMID:17537593

  7. 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

  8. 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

  9. 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. PMID:26913026

  10. Strain-specific regulation of intracellular Wolbachia density in multiply infected insects.

    PubMed

    Mouton, L; Henri, H; Bouletreau, M; Vavre, F

    2003-12-01

    Vertically transmitted symbionts suffer a severe reduction in numbers when they pass through host generations, resulting in genetic homogeneity or even clonality of their populations. Wolbachia endosymbionts that induce cytoplasmic incompatibility in their hosts depart from this rule, because cytoplasmic incompatibility actively maintains multiple infection within hosts. Hosts and symbionts are thus probably under peculiar selective pressures that must shape the way intracellular bacterial populations are regulated. We studied the density and location of Wolbachia within adult Leptopilina heterotoma, a haplodiploid wasp that is parasitic on Drosophila and that is naturally infected with three Wolbachia strains, but for which we also obtained one simply infected and two doubly infected lines. Comparison of these four lines by quantitative polymerase chain reaction using a real-time detection system showed that total Wolbachia density varies according to the infection status of individuals, while the specific density of each Wolbachia strain remains constant regardless of the presence of other strains. This suggests that Wolbachia strains do not compete with one another within the same host individual, and that a strain-specific regulatory mechanism is operating. We discuss the regulatory mechanisms that are involved, and how this process might have evolved as a response to selective pressures acting on both partners. PMID:14629360

  11. Detection and Localization of Wolbachia in Thrips palmi Karny (Thysanoptera: Thripidae).

    PubMed

    Saurav, Gunjan Kumar; Daimei, Guisuibou; Rana, Vipin Singh; Popli, Sonam; Rajagopal, Raman

    2016-06-01

    Thrips palmi Karny is a globally distributed polyphagous agricultural pest. It causes huge economic loss by its biological behaviors like feeding, reproduction and transmission of tospoviruses. Since T. palmi shows close morphological similarities with other thrips species, we employed mitochondrial cytochrome oxidase 1 (mtCO1) gene as a molecular marker. BLAST analysis of this sequence helped us to identify the collected specimen as T. palmi. We observed the female to male ratio of about 3:1 from collected samples and suspected the presence of Wolbachia. The presence of Wolbachia was detected by PCR using genus specific primers of 16S rRNA gene. Further confirmation of Wolbachia strain was achieved by conducting PCR amplification of three ubiquitous genes ftsZ, gatB and groEL. A phylogenetic tree was constructed with concatenated sequences of ftsZ and gatB gene to assign supergroup to Wolbachia. Finally, we localized Wolbachia in abdominal region of the insect using fluorescent in situ hybridization with the help of confocal microscope. Our result confirmed the presence of Wolbachia supergroup B strain for the first time in T. palmi. PMID:27570308

  12. Identification and characterization of Wolbachia in Solenopsis saevissima fire ants (Hymenoptera: Formicidae) in southeastern Brazil.

    PubMed

    Fernando de Souza, Rodrigo; Daivison Silva Ramalho, João; Santina de Castro Morini, Maria; Wolff, José Luiz Caldas; Araújo, Ronaldo Carvalho; Mascara, Douglas

    2009-03-01

    The genus Solenopsis appears to have evolved and radiated very rapidly in South America and then spread throughout the rest of the continent. As part of the expansion process, distribution patterns and different degrees of geographic isolation among populations of S. saevissima can be observed. We have investigated the presence of Wolbachia in 52 colonies and 1623 individuals in southeastern Brazil. Detection of Wolbachia infection was based on amplification of the 16S rRNA and wsp genes by polymerase chain reactions. Wolbachia was found in only one of the four locations investigated and it was observed that the populations were polymorphic for infection. The infection level observed increased during the period of screening. In particular, double infection (16SWA and B) increased from 44% in 2005 to 90% in 2006. The A-group of Wolbachia from the wsp sequences was determined by sequencing. However, two variant wsp sequences were detected in Wolbachia present in these populations. The alignment of our sequences with those deposited in GenBank indicated significant differences in relation to homologous sequences. Phylogenetic relationships were inferred using parsimony, and confidence intervals were estimated by bootstrapping. Then the divergence of the Wolbachia of S. saevissima in the populations studied with other variants allowed us to verify that wSS1 and dwSS2 formed a distinct clade within the A-group (>75%). These results can be useful in studies on the dynamics of ant populations. PMID:18979132

  13. 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

  14. The role of Wolbachia bacteria in reproductive incompatibilities and hybrid zones of Diabrotica beetles and Gryllus crickets.

    PubMed

    Giordano, R; Jackson, J J; Robertson, H M

    1997-10-14

    A rickettsial bacterium in the genus Wolbachia is the cause of a unidirectional reproductive incompatibility observed between two major beetle pests of maize, the western corn rootworm, Diabrotica virgifera virgifera, and the Mexican corn rootworm, D. v. zeae. These subspecies are allopatric except for two known regions of sympatry in Texas and Mexico. We demonstrate that populations of D. v. virgifera, with the exception of two populations in southern Arizona, are infected with a strain of Wolbachia. Populations of D. v. zeae are not infected. Treatment of D. v. virgifera with tetracycline eliminated the Wolbachia and removed the reproductive incompatibility. Similar patterns of reproductive incompatibility exist among taxa of the cricket genus Gryllus. Gryllus assimilis, G. integer, G. ovisopis, G. pennsylvanicus, and G. rubens are infected with Wolbachia whereas G. firmus is usually not. Populations of G. rubens and G. ovisopis carry the same Wolbachia strain, which is distinct from that of G. integer. G. pennsylvanicus is infected with two Wolbachia strains, that found in G. rubens and one unique to G. pennsylvanicus. Moreover, a proportion of G. pennsylvanicus individuals harbors both strains. Wolbachia may have influenced speciation in some members of the genus Gryllus by affecting the degree of hybridization between species. Given that Wolbachia infections are relatively common in insects, it is likely that other insect hybrid zones may be influenced by infections with Wolbachia. PMID:9326628

  15. The role of Wolbachia bacteria in reproductive incompatibilities and hybrid zones of Diabrotica beetles and Gryllus crickets

    PubMed Central

    Giordano, Rosanna; Jackson, Jan J.; Robertson, Hugh M.

    1997-01-01

    A rickettsial bacterium in the genus Wolbachia is the cause of a unidirectional reproductive incompatibility observed between two major beetle pests of maize, the western corn rootworm, Diabrotica virgifera virgifera, and the Mexican corn rootworm, D. v. zeae. These subspecies are allopatric except for two known regions of sympatry in Texas and Mexico. We demonstrate that populations of D. v. virgifera, with the exception of two populations in southern Arizona, are infected with a strain of Wolbachia. Populations of D. v. zeae are not infected. Treatment of D. v. virgifera with tetracycline eliminated the Wolbachia and removed the reproductive incompatibility. Similar patterns of reproductive incompatibility exist among taxa of the cricket genus Gryllus. Gryllus assimilis, G. integer, G. ovisopis, G. pennsylvanicus, and G. rubens are infected with Wolbachia whereas G. firmus is usually not. Populations of G. rubens and G. ovisopis carry the same Wolbachia strain, which is distinct from that of G. integer. G. pennsylvanicus is infected with two Wolbachia strains, that found in G. rubens and one unique to G. pennsylvanicus. Moreover, a proportion of G. pennsylvanicus individuals harbors both strains. Wolbachia may have influenced speciation in some members of the genus Gryllus by affecting the degree of hybridization between species. Given that Wolbachia infections are relatively common in insects, it is likely that other insect hybrid zones may be influenced by infections with Wolbachia. PMID:9326628

  16. Immunohistochemical/immunogold detection and distribution of the endosymbiont Wolbachia of Dirofilaria immitis and Brugia pahangi using a polyclonal antiserum raised against WSP (Wolbachia surface protein).

    PubMed

    Kramer, L H; Passeri, B; Corona, S; Simoncini, L; Casiraghi, M

    2003-03-01

    Intracellular bacteria in filarial nematodes were described as early as the 1970s, yet it was only with the work on Dirofilaria immitis, the agent of canine and feline heartworm disease, that these microorganisms were identified as belonging to Wolbachia, a genus known for encompassing bacteria infecting insects and other arthropods. The implications for the presence of intracellular bacteria in filarial nematodes is now the subject of intense research, particularly regarding their role in the immunology and pathogenesis of disease in infected humans and animals and as a possible target for therapy. Here, the authors report results on the immunohistochemical and immunogold staining of Wolbachia in D. immitis and Brugia pahangi using polyclonal antibodies raised against the recombinant Wolbachia surface protein (WSP). The bacteria were present in the lateral hypodermal chords of both male and female worms and in the reproductive tract of adult females (oocytes, morulae, microfilariae). In D. immitis and B. pahangi from animals treated with tetracycline, positive staining was observed in the lateral chords of adult males and females, but was absent from the oocytes and morulae. These results indicate that Wolbachia endosymbionts can be identified immunohistochemically with anti-WSP polyclonal antibodies, that their distribution matches that already described for Wolbachia of other filarial worms, and that antibiotic treatment may impede the vertical transmission of these bacteria. Unequivocal detection of Wolbachia is essential for the study of this symbiont, in particular to monitor the effects of antibiotic treatment on worms. The use of a specific marker for bacteria in their nematode hosts represents an extremely useful tool in evaluating the pathogenic role and the effect of antibiotic treatment on these potential targets in the control of filarial disease. PMID:12632152

  17. 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.

  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. Tandem repeat markers as novel diagnostic tools for high resolution fingerprinting of Wolbachia

    PubMed Central

    2012-01-01

    Background Strains of the endosymbiotic bacterium Wolbachia pipientis are extremely diverse both genotypically and in terms of their induced phenotypes in invertebrate hosts. Despite extensive molecular characterisation of Wolbachia diversity, little is known about the actual genomic diversity within or between closely related strains that group tightly on the basis of existing gene marker systems, including Multiple Locus Sequence Typing (MLST). There is an urgent need for higher resolution fingerprinting markers of Wolbachia for studies of population genetics, horizontal transmission and experimental evolution. Results The genome of the wMel Wolbachia strain that infects Drosophila melanogaster contains inter- and intragenic tandem repeats that may evolve through expansion or contraction. We identified hypervariable regions in wMel, including intergenic Variable Number Tandem Repeats (VNTRs), and genes encoding ankyrin (ANK) repeat domains. We amplified these markers from 14 related Wolbachia strains belonging to supergroup A and were successful in differentiating size polymorphic alleles. Because of their tandemly repeated structure and length polymorphism, the markers can be used in a PCR-diagnostic multilocus typing approach, analogous to the Multiple Locus VNTR Analysis (MLVA) established for many other bacteria and organisms. The isolated markers are highly specific for supergroup A and not informative for other supergroups. However, in silico analysis of completed genomes from other supergroups revealed the presence of tandem repeats that are variable and could therefore be useful for typing target strains. Conclusions Wolbachia genomes contain inter- and intragenic tandem repeats that evolve through expansion or contraction. A selection of polymorphic tandem repeats is a novel and useful PCR diagnostic extension to the existing MLST typing system of Wolbachia, as it allows rapid and inexpensive high-throughput fingerprinting of closely related strains for

  20. The Wolbachia Genome of Brugia malayi: Endosymbiont Evolution within a Human Pathogenic Nematode

    PubMed Central

    2005-01-01

    Complete genome DNA sequence and analysis is presented for Wolbachia, the obligate alpha-proteobacterial endosymbiont required for fertility and survival of the human filarial parasitic nematode Brugia malayi. Although, quantitatively, the genome is even more degraded than those of closely related Rickettsia species, Wolbachia has retained more intact metabolic pathways. The ability to provide riboflavin, flavin adenine dinucleotide, heme, and nucleotides is likely to be Wolbachia's principal contribution to the mutualistic relationship, whereas the host nematode likely supplies amino acids required for Wolbachia growth. Genome comparison of the Wolbachia endosymbiont of B. malayi (wBm) with the Wolbachia endosymbiont of Drosophila melanogaster (wMel) shows that they share similar metabolic trends, although their genomes show a high degree of genome shuffling. In contrast to wMel, wBm contains no prophage and has a reduced level of repeated DNA. Both Wolbachia have lost a considerable number of membrane biogenesis genes that apparently make them unable to synthesize lipid A, the usual component of proteobacterial membranes. However, differences in their peptidoglycan structures may reflect the mutualistic lifestyle of wBm in contrast to the parasitic lifestyle of wMel. The smaller genome size of wBm, relative to wMel, may reflect the loss of genes required for infecting host cells and avoiding host defense systems. Analysis of this first sequenced endosymbiont genome from a filarial nematode provides insight into endosymbiont evolution and additionally provides new potential targets for elimination of cutaneous and lymphatic human filarial disease. PMID:15780005

  1. 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

  2. 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. PMID:25671520

  3. 33 CFR 104.106 - Passenger access area.

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... measures for access control, of a ferry, passenger vessel, or cruise ship that is open to passengers. It is... 33 Navigation and Navigable Waters 1 2014-07-01 2014-07-01 false Passenger access area. 104.106... SECURITY MARITIME SECURITY: VESSELS General § 104.106 Passenger access area. (a) A ferry, passenger...

  4. 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:...

  5. 33 CFR 104.106 - Passenger access area.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... measures for access control, of a ferry, passenger vessel, or cruise ship that is open to passengers. It is... 33 Navigation and Navigable Waters 1 2010-07-01 2010-07-01 false Passenger access area. 104.106... SECURITY MARITIME SECURITY: VESSELS General § 104.106 Passenger access area. (a) A ferry, passenger...

  6. 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.

  7. Locating influential nodes via dynamics-sensitive centrality

    NASA Astrophysics Data System (ADS)

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

    2016-02-01

    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.

  8. 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. PMID:17340854

  9. 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...

  10. 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

  11. 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. PMID:22188300

  12. TWO DIFFERENT WOLBACHIA TYPES CREATE A BREEDING BARRIER BETWEEN POPULATIONS OF NORTHERN CORN ROOTWORM (DIABROTICA BARBERI) IN ILLINOIS

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Wolbachia sp. are endosymbiotic bacteria that are widespread in Arthropods and are often associated with reproductive incompatibilities between infected and uninfected individuals. 16S rDNA primers detected Wolbachia in northern corn rootworms (NCR) from the eastern portion of their range with a W...

  13. 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...

  14. [Medical problems among airline passengers].

    PubMed

    Owe, J O; Christensen, C C

    1998-09-30

    Worldwide, there are more than one billion air travelers each year. Flying in a modern jet airliner is a safe, efficient and relatively comfortable mode of transport, although a few susceptible passengers may be adversely affected by environmental and physiological stresses like pressure change, reduced level of oxygen, dry air, immobility due to cramped seating, noise, vibration and turbulence, in addition to stressful airports. This article describes these factors and their medical implications and includes some practical medical advice to travellers. Reported inflight illness and injuries in two major Scandinavian airlines 1993-97 are presented. PMID:9820008

  15. Male-killing Wolbachia and mitochondrial selective sweep in a migratory African insect

    PubMed Central

    2012-01-01

    Background Numerous recent studies have shown that resident symbiotic microorganisms of insects play a fundamental role in host ecology and evolution. The lepidopteran pest, African armyworm (Spodoptera exempta), is a highly migratory and destructive species found throughout sub-Saharan Africa, that can experience eruptive outbreaks within the space of a single generation, making predicting population dynamics and pest control forecasting extremely difficult. Three strains of Wolbachia have recently been identified infecting this species in populations sampled from Tanzania. In this study, we examined the interaction between Wolbachia pipiensis infections and the co-inherited marker, mtDNA, within populations of armyworm, as a means to investigate the population biology and evolutionary history of Wolbachia and its host. Results A Wolbachia-infected isofemale line was established in the laboratory. Phenotypic studies confirmed the strain wExe1 as a male-killer. Partial sequencing of the mitochondrial COI gene from 164 individual field-collected armyworm of known infection status revealed 17 different haplotypes. There was a strong association between Wolbachia infection status and mtDNA haplotype, with a single dominant haplotype, haplo1 (90.2% prevalence), harbouring the endosymbiont. All three Wolbachia strains were associated with this haplotype. This indicates that Wolbachia may be driving a selective sweep on armyworm haplotype diversity. Despite very strong biological and molecular evidence that the samples represent a single species (including from nuclear 28S gene markers), the 17 haplotypes did not fall into a monophyletic clade within the Spodoptera genus; with six haplotypes (2 each from 3 geographically separate populations) differing by >11% in their nucleotide sequence to the other eleven. Conclusions This study suggests that three strains of Wolbachia may be driving a selective sweep on armyworm haplotype diversity, and that based on COI sequence

  16. Population genetic structure and Wolbachia infection in an endangered butterfly, Zizina emelina (Lepidoptera, Lycaenidae), in Japan.

    PubMed

    Sakamoto, Y; Hirai, N; Tanikawa, T; Yago, M; Ishii, M

    2015-04-01

    Zizina emelina (de l'Orza) is listed on Japan's Red Data List as an endangered species because of loss of its principal food plant and habitat. We compared parts of the mitochondrial and nuclear genes of this species to investigate the level of genetic differentiation among the 14 extant populations. We also examined infection of the butterfly with the bacterium Wolbachia to clarify the bacterium's effects on the host population's genetic structure. Mitochondrial and nuclear DNA analyses revealed that haplotype composition differed significantly among most of the populations, and the fixation index F ST was positively correlated with geographic distance. In addition, we found three strains of Wolbachia, one of which was a male killer; these strains were prevalent in several populations. There was linkage between some host mitochondrial haplotypes and the three Wolbachia strains, although no significant differences were found in a comparison of host mitochondrial genetic diversity with nuclear genetic diversity in Wolbachia-infected or -uninfected populations. These genetic analyses and Wolbachia infection findings show that Z. emelina has little migratory activity and that little gene flow occurs among the current populations. PMID:25499047

  17. 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

  18. Wolbachia Infection and Resource Competition Effects on Immature Aedes albopictus (Diptera: Culicidae)

    PubMed Central

    Gavotte, Laurent; Mercer, David R.; Vandyke, Rhonda; Mains, James W.; Dobson, Stephen L.

    2009-01-01

    Wolbachia pipientis Hertig and Wolbach (Rickettsiales: Rickettsiaceae) are intracellular α-proteobacteria that occur naturally in Aedes albopictus (Skuse) (Diptera: Culicidae) and numerous other invertebrates. These endosymbionts can invade host populations by manipulating host reproduction. Wolbachia infections have been shown to impart both costs and benefits to hosts in terms of development, survival, and fecundity. Here, we monitor intraspecific competition among independent cohorts of infected or uninfected larvae. Levels of competition are manipulated by varying initial larval densities and food levels. Although larval density is observed to have major impacts on immature survivorship, sex ratio of eclosing adults, and developmental rates, the Wolbachia infection status had minimal impact on male immatures and no effect on immature females under these experimental conditions. Female and male immatures were observed to respond differently to competitive pressure, with the functional relationships of females and males consistent with scramble and contest competition, respectively. The results are discussed in relation to the evolution of naturally occurring Wolbachia infections in Ae. albopictus (i.e., natural population replacement events) and public health strategies that propose the manipulation of Wolbachia infections in Ae. albopictus populations. PMID:19496412

  19. Strain-specific quantification of Wolbachia density in Aedes albopictus and effects of larval rearing conditions.

    PubMed

    Dutton, T J; Sinkins, S P

    2004-06-01

    The density of the endosymbiont Wolbachia can influence the expression of the crossing sterilities known as cytoplasmic incompatibility (CI), and also its rate of maternal transmission. Aedes albopictus mosquitoes contain a superinfection with the Wolbachia strains wAlbA and wAlbB. A strain-specific real-time quantitative PCR assay was developed and used to quantify relative Wolbachia strain densities within individual mosquitoes. The wAlbB strain was consistently found to be at higher density than wAlbA, which can explain a slightly lower rate of maternal transmission reported for wAlbA. The effects of larval crowding and nutritional stress were also examined. Larval crowding always reduced adult size, but reduced the density of Wolbachia strains relative to uncrowded conditions only if crowding was accompanied by restricted nutrient availability. Crowded rearing conditions never resulted in strain segregation or in a reduction in the penetrance of CI, however. The rate of maternal transmission and the penetrance of CI are the two most important variables that determine relative Wolbachia population invasion dynamics, and both are considerably higher here than have been reported in the Drosophila simulans model system. PMID:15157232

  20. Variability and Expression of Ankyrin Domain Genes in Wolbachia Variants Infecting the Mosquito Culex pipiens▿ †

    PubMed Central

    Duron, Olivier; Boureux, Anthony; Echaubard, Pierre; Berthomieu, Arnaud; Berticat, Claire; Fort, Philippe; Weill, Mylène

    2007-01-01

    Wolbachia strains are maternally inherited endosymbiotic bacteria that infect many arthropod species and have evolved several different ways of manipulating their hosts, the most frequent way being cytoplasmic incompatibility (CI). CI leads to embryo death in crosses between infected males and uninfected females as well as in crosses between individuals infected by incompatible Wolbachia strains. The mosquito Culex pipiens exhibits the highest crossing type variability reported so far. Our crossing data support the notion that CI might be driven by at least two distinct genetic units that control the CI functions independently in males and females. Although the molecular basis of CI remains unknown, proteins with ankyrin (ANK) domains represent promising candidates since they might interact with a wide range of host proteins. Here we searched for sequence variability in the 58 ANK genes carried in the genomes of Wolbachia variants infecting Culex pipiens. Only five ANK genes were polymorphic in the genomes of incompatible Wolbachia variants, and none correlated with the CI pattern obtained with 15 mosquito strains (representing 14 Wolbachia variants). Further analysis of ANK gene expression evidenced host- and sex-dependent variations, which did not improve the correlation. Taken together, these data do not support the direct implication of ANK genes in CI determinism. PMID:17449622

  1. Effect of temperature on Wolbachia density and impact on cytoplasmic incompatibility.

    PubMed

    Mouton, L; Henri, H; Bouletreau, M; Vavre, F

    2006-01-01

    The outcome and the evolution of host-symbiont associations depend on environmental constraints, but responses are difficult to predict since they arise from a complex interaction between the host, the parasite and the environment. The situation can be even more complex when multiple parasite genotypes, with potentially different responses to environmental changes, coexist within a single host. In this paper, we investigated the effect of the temperature (from 14 to 26 degrees C) during the host development on the density of 3 strains of the intracellular bacterium Wolbachia that coexist within the wasp Leptopilina heterotoma. In this species, Wolbachia induces cytoplasmic incompatibility, a sperm-egg incompatibility that allows it to spread and persist in host populations. Using real-time quantitative PCR we found that (i) Wolbachia density is temperature-specific and highest at 26 degrees C; (ii) the order of the abundance of the 3 Wolbachia strains does not vary with temperature changes; (iii) the response of bacterial density to temperature occurs within a single insect generation, during the egg-to-adult developmental period; (iv) in this species, temperature-related changes in Wolbachia density do not influence cytoplasmic incompatibility. PMID:16393353

  2. 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

  3. 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. PMID:27012719

  4. 14 CFR 23.1524 - Maximum passenger seating configuration.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... 14 Aeronautics and Space 1 2012-01-01 2012-01-01 false Maximum passenger seating configuration. 23... Operating Limitations and Information § 23.1524 Maximum passenger seating configuration. The maximum passenger seating configuration must be established....

  5. 14 CFR 23.1524 - Maximum passenger seating configuration.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... 14 Aeronautics and Space 1 2011-01-01 2011-01-01 false Maximum passenger seating configuration. 23... Operating Limitations and Information § 23.1524 Maximum passenger seating configuration. The maximum passenger seating configuration must be established....

  6. 14 CFR 23.1524 - Maximum passenger seating configuration.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... 14 Aeronautics and Space 1 2013-01-01 2013-01-01 false Maximum passenger seating configuration. 23... Operating Limitations and Information § 23.1524 Maximum passenger seating configuration. The maximum passenger seating configuration must be established....

  7. 14 CFR 23.1524 - Maximum passenger seating configuration.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 14 Aeronautics and Space 1 2010-01-01 2010-01-01 false Maximum passenger seating configuration. 23... Operating Limitations and Information § 23.1524 Maximum passenger seating configuration. The maximum passenger seating configuration must be established....

  8. 14 CFR 23.1524 - Maximum passenger seating configuration.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... 14 Aeronautics and Space 1 2014-01-01 2014-01-01 false Maximum passenger seating configuration. 23... Operating Limitations and Information § 23.1524 Maximum passenger seating configuration. The maximum passenger seating configuration must be established....

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

    PubMed Central

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

    2015-01-01

    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. PMID:25059495

  10. 14 CFR 136.7 - Passenger briefings.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 14 Aeronautics and Space 3 2010-01-01 2010-01-01 false Passenger briefings. 136.7 Section 136.7... PARKS AIR TOUR MANAGEMENT National Air Tour Safety Standards § 136.7 Passenger briefings. (a) Before... opening exits and exiting the aircraft. (b) For flight segments over water beyond the shoreline,...

  11. 46 CFR 115.113 - Passengers permitted.

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ...) of rail space available to the passengers at the periphery of each deck. The following rail space may not be used in determining the maximum number of passengers permitted: (i) Rail space in congested... sail booms, running rigging, or paddle wheels, or along pulpits; (ii) Rail space on stairways; and...

  12. 78 FR 49248 - Passenger Vessels Accessibility Guidelines

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-08-13

    ... additional time to submit comments. DATES: For the proposed rule published June 25, 2013 (78 FR 38102... passengers with disabilities. See 78 FR 38102, June 25, 2013. In that notice, the Access Board requested... TRANSPORTATION BARRIERS COMPLIANCE BOARD 36 CFR Part 1196 RIN 3014-AA11 Passenger Vessels...

  13. Passenger and Naturalization Lists: The New Sources.

    ERIC Educational Resources Information Center

    Filby, P. William

    1983-01-01

    Reviews information sources designed to assist the genealogical researcher with the arrival of his/her ancestors: "A Bibliography of Ship Passenger Lists 1538-1825"; "Passenger and Immigration Lists Index"; "Philadelphia Naturalization Records." Examples provided include name entry, source citation, annotation, and subject entries. Nineteen…

  14. 14 CFR 91.519 - Passenger briefing.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 14 Aeronautics and Space 2 2010-01-01 2010-01-01 false Passenger briefing. 91.519 Section 91.519 Aeronautics and Space FEDERAL AVIATION ADMINISTRATION, DEPARTMENT OF TRANSPORTATION (CONTINUED) AIR TRAFFIC... be supplemented by printed cards for the use of each passenger containing— (1) A diagram of,...

  15. Genomics of Loa loa, a Wolbachia-free filarial parasite of humans.

    PubMed

    Desjardins, Christopher A; Cerqueira, Gustavo C; Goldberg, Jonathan M; Dunning Hotopp, Julie C; Haas, Brian J; Zucker, Jeremy; Ribeiro, José M C; Saif, Sakina; Levin, Joshua Z; Fan, Lin; Zeng, Qiandong; Russ, Carsten; Wortman, Jennifer R; Fink, Doran L; Birren, Bruce W; Nutman, Thomas B

    2013-05-01

    Loa loa, the African eyeworm, is a major filarial pathogen of humans. Unlike most filariae, L. loa does not contain the obligate intracellular Wolbachia endosymbiont. We describe the 91.4-Mb genome of L. loa and that of the related filarial parasite Wuchereria bancrofti and predict 14,907 L. loa genes on the basis of microfilarial RNA sequencing. By comparing these genomes to that of another filarial parasite, Brugia malayi, and to those of several other nematodes, we demonstrate synteny among filariae but not with nonparasitic nematodes. The L. loa genome encodes many immunologically relevant genes, as well as protein kinases targeted by drugs currently approved for use in humans. Despite lacking Wolbachia, L. loa shows no new metabolic synthesis or transport capabilities compared to other filariae. These results suggest that the role of Wolbachia in filarial biology is more subtle than previously thought and reveal marked differences between parasitic and nonparasitic nematodes. PMID:23525074

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

    PubMed Central

    Fallon, Ann M.

    2016-01-01

    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. PMID:26019119

  17. 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. PMID:22669278

  18. Finding Influential Spreaders from Human Activity beyond Network Location

    PubMed Central

    Min, Byungjoon; Liljeros, Fredrik; Makse, Hernán A.

    2015-01-01

    Most centralities proposed for identifying influential spreaders on social networks to either spread a message or to stop an epidemic require the full topological information of the network on which spreading occurs. In practice, however, collecting all connections between agents in social networks can be hardly achieved. As a result, such metrics could be difficult to apply to real social networks. Consequently, a new approach for identifying influential people without the explicit network information is demanded in order to provide an efficient immunization or spreading strategy, in a practical sense. In this study, we seek a possible way for finding influential spreaders by using the social mechanisms of how social connections are formed in real networks. We find that a reliable immunization scheme can be achieved by asking people how they interact with each other. From these surveys we find that the probabilistic tendency to connect to a hub has the strongest predictive power for influential spreaders among tested social mechanisms. Our observation also suggests that people who connect different communities is more likely to be an influential spreader when a network has a strong modular structure. Our finding implies that not only the effect of network location but also the behavior of individuals is important to design optimal immunization or spreading schemes. PMID:26323015

  19. Finding Influential Spreaders from Human Activity beyond Network Location.

    PubMed

    Min, Byungjoon; Liljeros, Fredrik; Makse, Hernán A

    2015-01-01

    Most centralities proposed for identifying influential spreaders on social networks to either spread a message or to stop an epidemic require the full topological information of the network on which spreading occurs. In practice, however, collecting all connections between agents in social networks can be hardly achieved. As a result, such metrics could be difficult to apply to real social networks. Consequently, a new approach for identifying influential people without the explicit network information is demanded in order to provide an efficient immunization or spreading strategy, in a practical sense. In this study, we seek a possible way for finding influential spreaders by using the social mechanisms of how social connections are formed in real networks. We find that a reliable immunization scheme can be achieved by asking people how they interact with each other. From these surveys we find that the probabilistic tendency to connect to a hub has the strongest predictive power for influential spreaders among tested social mechanisms. Our observation also suggests that people who connect different communities is more likely to be an influential spreader when a network has a strong modular structure. Our finding implies that not only the effect of network location but also the behavior of individuals is important to design optimal immunization or spreading schemes. PMID:26323015

  20. 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

  1. Incidence of Wolbachia and Cardinium endosymbionts in the Osmia community in Korea.

    PubMed

    Jeong, Gilsang; Lee, Kyeongyong; Choi, Jiyoung; Hwang, Seokjo; Park, Byeongdo; Kim, Wontae; Choi, Youngcheol; Park, Ingyun; Kim, Jonggill

    2009-02-01

    Sex ratio distorting endosymbionts induce reproductive anomalies in their arthropod hosts. They have recently been paid much attention as firstly texts of evolution of host-symbiont relationships and secondly potential biological control agents to control arthropod pests. Among such organisms, Wolbachia and Cardinium bacteria are well characterized. This study aims at probing such bacteria in the Osmia community to evaluate their potential utilization to control arthropod pests. Among 17 PCR tested species, Osmia cornifrons and a parasitic fly are infected with Wolbachia and a mite species is infected with Cardinium. Phylogenetic tree analyses suggest that horizontal transfer of the bacteria occurred between phylogenetically distant hosts. PMID:19229488

  2. Wolbachia symbiont infections induce strong cytoplasmic incompatibility in the tsetse fly Glossina morsitans.

    PubMed

    Alam, Uzma; Medlock, Jan; Brelsfoard, Corey; Pais, Roshan; Lohs, Claudia; Balmand, Séverine; Carnogursky, Jozef; Heddi, Abdelaziz; Takac, Peter; Galvani, Alison; Aksoy, Serap

    2011-12-01

    Tsetse flies are vectors of the protozoan parasite African trypanosomes, which cause sleeping sickness disease in humans and nagana in livestock. Although there are no effective vaccines and efficacious drugs against this parasite, vector reduction methods have been successful in curbing the disease, especially for nagana. Potential vector control methods that do not involve use of chemicals is a genetic modification approach where flies engineered to be parasite resistant are allowed to replace their susceptible natural counterparts, and Sterile Insect technique (SIT) where males sterilized by chemical means are released to suppress female fecundity. The success of genetic modification approaches requires identification of strong drive systems to spread the desirable traits and the efficacy of SIT can be enhanced by identification of natural mating incompatibility. One such drive mechanism results from the cytoplasmic incompatibility (CI) phenomenon induced by the symbiont Wolbachia. CI can also be used to induce natural mating incompatibility between release males and natural populations. Although Wolbachia infections have been reported in tsetse, it has been a challenge to understand their functional biology as attempts to cure tsetse of Wolbachia infections by antibiotic treatment damages the obligate mutualistic symbiont (Wigglesworthia), without which the flies are sterile. Here, we developed aposymbiotic (symbiont-free) and fertile tsetse lines by dietary provisioning of tetracycline supplemented blood meals with yeast extract, which rescues Wigglesworthia-induced sterility. Our results reveal that Wolbachia infections confer strong CI during embryogenesis in Wolbachia-free (Gmm(Apo)) females when mated with Wolbachia-infected (Gmm(Wt)) males. These results are the first demonstration of the biological significance of Wolbachia infections in tsetse. Furthermore, when incorporated into a mathematical model, our results confirm that Wolbachia can be used

  3. Genome Evolution of Wolbachia Strain wPip from the Culex pipiens Group

    PubMed Central

    Klasson, Lisa; Walker, Thomas; Sebaihia, Mohammed; Sanders, Mandy J.; Quail, Michael A.; Lord, Angela; Sanders, Susanne; Earl, Julie; O'Neill, Scott L.; Thomson, Nicholas; Sinkins, Steven P.; Parkhill, Julian

    2008-01-01

    The obligate intracellular bacterium Wolbachia pipientis strain wPip induces cytoplasmic incompatibility (CI), patterns of crossing sterility, in the Culex pipiens group of mosquitoes. The complete sequence is presented of the 1.48-Mbp genome of wPip which encodes 1386 coding sequences (CDSs), representing the first genome sequence of a B-supergroup Wolbachia. Comparisons were made with the smaller genomes of Wolbachia strains wMel of Drosophila melanogaster, an A-supergroup Wolbachia that is also a CI inducer, and wBm, a mutualist of Brugia malayi nematodes that belongs to the D-supergroup of Wolbachia. Despite extensive gene order rearrangement, a core set of Wolbachia genes shared between the 3 genomes can be identified and contrasts with a flexible gene pool where rapid evolution has taken place. There are much more extensive prophage and ankyrin repeat encoding (ANK) gene components of the wPip genome compared with wMel and wBm, and both are likely to be of considerable importance in wPip biology. Five WO-B–like prophage regions are present and contain some genes that are identical or highly similar in multiple prophage copies, whereas other genes are unique, and it is likely that extensive recombination, duplication, and insertion have occurred between copies. A much larger number of genes encode ankyrin repeat (ANK) proteins in wPip, with 60 present compared with 23 in wMel, many of which are within or close to the prophage regions. It is likely that this pattern is partly a result of expansions in the wPip lineage, due for example to gene duplication, but their presence is in some cases more ancient. The wPip genome underlines the considerable evolutionary flexibility of Wolbachia, providing clear evidence for the rapid evolution of ANK-encoding genes and of prophage regions. This host–Wolbachia system, with its complex patterns of sterility induced between populations, now provides an excellent model for unraveling the molecular systems underlying host

  4. 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.

  5. Culture and Phenotypic Characterization of a Wolbachia pipientis Isolate

    PubMed Central

    Fenollar, Florence; La Scola, Bernard; Inokuma, Hisashi; Dumler, J. Stephen; Taylor, Mark J.; Raoult, Didier

    2003-01-01

    The recent isolation of Wolbachia pipientis in the continuous cell line Aa23, established from eggs of a strain of the Asian tiger mosquito Aedes albopictus, allowed us to perform extensive characterization of the isolate. Bacterial growth could be obtained in C6/36, another A. albopictus cell line, at 28°C and in a human embryonic lung fibroblast monolayer at 28 and 37°C, confirming that its host cell range is broader than was initially thought. The bacteria were best visualized by Diff-Quik and May-Grünwald-Giemsa staining. Proteins from 213 to 18 kDa with two major protein bands of 65 and 25 kDa were observed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. By Western blotting with specific polyclonal mouse and rabbit antisera, dominant immunoreactive antigens were found at approximately 100, 80, and 30 kDa. The genome size was calculated to be 1,790 ± 17 kb by pulsed-field gel electrophoresis. The sequence of the citrate synthase gene (gltA) of W. pipientis was determined by gene walking. Its position in the phylogenetic tree constructed with gltA confirmed that found in a phylogenetic tree constructed with 16S rRNA genes and that it belongs in the α subgroup of the class Proteobacteria and that it is closely related to but independent from the genera Ehrlichia, Anaplasma, and Neorickettsia. PMID:14662922

  6. Low speed vehicle passenger ejection restraint effectiveness.

    PubMed

    Seluga, Kristopher J; Ojalvo, Irving U; Obert, Richard M

    2005-07-01

    Current golf carts and LSV's (Low Speed Vehicles) produce a significant number of passenger ejections during sharp turns. These LSV's do not typically possess seatbelts, but do provide outboard bench seat hip restraints that also serve as handholds. However, many current restraint designs appear incapable of preventing passenger ejections due to their low height and inefficient handhold position. Alternative handhold and hip restraint designs may improve passenger safety. Accordingly, this paper examines minimum size requirements for hip restraints to prevent passenger ejection during sharp turns and evaluates the effectiveness of a handhold mounted at the center of the bench seat. In this study, a simulation of a turning cart supplies the dynamic input to a biomechanical model of an adult male seated in a golf cart. Various restraint combinations are considered, both with and without the central handhold, to determine the likelihood of passenger ejection. It is shown that only the largest restraint geometries prevent passenger ejection. Adequate hip restraints should be much larger than current designs and a central handhold should be provided. In this way, golf cart and LSV manufacturers could reduce passenger ejections and improve fleet safety by incorporating recommendations provided herein. PMID:15893288

  7. 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. PMID:25681033

  8. Anti-filarial Activity of Antibiotic Therapy Is Due to Extensive Apoptosis after Wolbachia Depletion from Filarial Nematodes

    PubMed Central

    Sullivan, William; Taylor, Mark J.

    2011-01-01

    Filarial nematodes maintain a mutualistic relationship with the endosymbiont Wolbachia. Depletion of Wolbachia produces profound defects in nematode development, fertility and viability and thus has great promise as a novel approach for treating filarial diseases. However, little is known concerning the basis for this mutualistic relationship. Here we demonstrate using whole mount confocal microscopy that an immediate response to Wolbachia depletion is extensive apoptosis in the adult germline, and in the somatic cells of the embryos, microfilariae and fourth-stage larvae (L4). Surprisingly, apoptosis occurs in the majority of embryonic cells that had not been infected prior to antibiotic treatment. In addition, no apoptosis occurs in the hypodermal chords, which are populated with large numbers of Wolbachia, although disruption of the hypodermal cytoskeleton occurs following their depletion. Thus, the induction of apoptosis upon Wolbachia depletion is non-cell autonomous and suggests the involvement of factors originating from Wolbachia in the hypodermal chords. The pattern of apoptosis correlates closely with the nematode tissues and processes initially perturbed following depletion of Wolbachia, embryogenesis and long-term sterilization, which are sustained for several months until the premature death of the adult worms. Our observations provide a cellular mechanism to account for the sustained reductions in microfilarial loads and interruption of transmission that occurs prior to macrofilaricidal activity following antibiotic therapy of filarial nematodes. PMID:22072969

  9. Persistence of a Wolbachia infection frequency cline in Drosophila melanogaster and the possible role of reproductive dormancy.

    PubMed

    Kriesner, Peter; Conner, William R; Weeks, Andrew R; Turelli, Michael; Hoffmann, Ary A

    2016-05-01

    Field populations of arthropods are often polymorphic for Wolbachia but the factors maintaining intermediate Wolbachia frequencies are generally not understood. In Drosophila melanogaster, Wolbachia frequencies are highly variable across the globe. We document the persistence of a Wolbachia infection frequency cline in D. melanogaster populations from eastern Australia across at least 20 years, with frequencies generally high in the tropics but lower in cool temperate regions. The results are interpreted using a model of frequency dynamics incorporating cytoplasmic incompatibility (CI), imperfect maternal transmission and Wolbachia effects on fitness. Clinal variation is less pronounced in eastern North America which may reflect annual recolonization at higher latitudes. Limited samples from Africa from latitudes matching our tropical and subtropical samples from Australia and North America show comparably high infection frequencies, but some equatorial samples show lower frequencies. Adult dormancy across cold periods may contribute to the Australian Wolbachia cline. Infected flies exposed to cold conditions for an extended period had reduced fecundity and viability, an effect not evident in unexposed controls. These fitness costs may contribute to the relatively low Wolbachia frequencies in Australian temperate areas; whereas different processes, including CI induced by young males, may contribute to higher frequencies in tropical locations. PMID:27076356

  10. Locating influential nodes via dynamics-sensitive centrality.

    PubMed

    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

  11. 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

  12. Comparison of airline passenger oxygen systems.

    PubMed

    Byrne, N J

    1995-08-01

    The principal sources of oxygen for inflight passenger use, scheduled and unscheduled, are examined. Present practices of assessment of the passenger's "fitness to fly" are described. Three partner airlines, British Airways, U.S. Air, and Qantas, catering for more than 8000 oxygen requests annually, are compared. Analysis of customer use suggests that medical oxygen requests are frequently not clinically justified. The growth in demand, for both scheduled and unscheduled use of an expensive resource, supports the need for a "recommended best practice" among carriers. Passengers with respiratory disorders who will most benefit from inflight oxygen are vulnerable either to hypoxia or asthma. PMID:7487813

  13. Effects of Host Sex, Plant Species, and Putative Host Species on the Prevalence of Wolbachia in Natural Populations of Bemisia tabaci (Hemiptera: Aleyrodidae): A Modified Nested PCR Study.

    PubMed

    Ji, Han-Le; Qi, Lan-Da; Hong, Xiao-Yue; Xie, Hong-Fang; Li, Yuan-Xi

    2015-02-01

    Bemisia tabaci (Gennadius) (Hemiptera: Aleyrodidae) is a globally distributed pest. One of the key endosymbionts in B. tabaci is Wolbachia, an α-proteobacterium implicated in many important biological processes. Previous studies indicated that the infection frequency of Wolbachia in Middle East-Asia Minor 1 (MEAM1) and Mediterranean (MED) varied greatly among populations in different areas. However, little is known about the factors that influence the prevalence of Wolbachia in B. tabaci. In this paper, 25 field populations were collected from different locations in China, and 1,161 individuals were screened for the presence of Wolbachia using a nested polymerase chain reaction (PCR)-based method, which targets the wsp gene, to confirm Wolbachia infection status. The prevalence of Wolbachia ranged from 1.54 to 66.67% within the 25 field populations, and the infection frequency of Wolbachia was affected significantly by the putative species of B. tabaci. The infection frequency (51.55%) of Wolbachia was significantly greater in native species than in the MED (25.65%) and MEAM1 (14.37%). With the exception of host plant, all factors, including putative species, geographic location, and the sex of the host, affected the Wolbachia infection frequency in whiteflies. Six Wolbachia strains were found and clustered into four distinct clades upon phylogenetic analyses. Furthermore, Wolbachia in B. tabaci have close relationships with those from other host species, including Liriomyza trifolii (Burgess), Sogatella furcifera (Horvath), Nilaparvata lugens (Stål), and Culex pipiens L. The results demonstrated the variation and diversity of Wolbachia in B. tabaci field populations, and that the application of nested PCR extended our knowledge of Wolbachia infection in B. tabaci, especially in invasive whiteflies. PMID:26470122

  14. 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...

  15. 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...

  16. 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...

  17. 46 CFR 176.910 - Passenger Ship Safety Certificate.

    Code of Federal Regulations, 2012 CFR

    2012-10-01

    ... 46 Shipping 7 2012-10-01 2012-10-01 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...

  18. 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...

  19. 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...

  20. 19 CFR 122.88 - Aircraft carrying domestic (stopover) passengers.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 19 Customs Duties 1 2012-04-01 2012-04-01 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...

  1. 19 CFR 122.88 - Aircraft carrying domestic (stopover) passengers.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 19 Customs Duties 1 2014-04-01 2014-04-01 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...

  2. 46 CFR 122.502 - Crew and passenger list.

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ... 46 Shipping 4 2011-10-01 2011-10-01 false Crew and passenger list. 122.502 Section 122.502 Shipping COAST GUARD, DEPARTMENT OF HOMELAND SECURITY (CONTINUED) SMALL PASSENGER VESSELS CARRYING MORE THAN 150 PASSENGERS OR WITH OVERNIGHT ACCOMMODATIONS FOR MORE THAN 49 PASSENGERS...

  3. 46 CFR 122.502 - Crew and passenger list.

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ... 46 Shipping 4 2013-10-01 2013-10-01 false Crew and passenger list. 122.502 Section 122.502 Shipping COAST GUARD, DEPARTMENT OF HOMELAND SECURITY (CONTINUED) SMALL PASSENGER VESSELS CARRYING MORE THAN 150 PASSENGERS OR WITH OVERNIGHT ACCOMMODATIONS FOR MORE THAN 49 PASSENGERS...

  4. 46 CFR 122.502 - Crew and passenger list.

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    ... 46 Shipping 4 2014-10-01 2014-10-01 false Crew and passenger list. 122.502 Section 122.502 Shipping COAST GUARD, DEPARTMENT OF HOMELAND SECURITY (CONTINUED) SMALL PASSENGER VESSELS CARRYING MORE THAN 150 PASSENGERS OR WITH OVERNIGHT ACCOMMODATIONS FOR MORE THAN 49 PASSENGERS...

  5. 46 CFR 122.502 - Crew and passenger list.

    Code of Federal Regulations, 2012 CFR

    2012-10-01

    ... 46 Shipping 4 2012-10-01 2012-10-01 false Crew and passenger list. 122.502 Section 122.502 Shipping COAST GUARD, DEPARTMENT OF HOMELAND SECURITY (CONTINUED) SMALL PASSENGER VESSELS CARRYING MORE THAN 150 PASSENGERS OR WITH OVERNIGHT ACCOMMODATIONS FOR MORE THAN 49 PASSENGERS...

  6. 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 Shipping COAST GUARD, DEPARTMENT OF HOMELAND SECURITY (CONTINUED) SMALL PASSENGER VESSELS CARRYING MORE THAN 150 PASSENGERS OR WITH OVERNIGHT ACCOMMODATIONS FOR MORE THAN 49 PASSENGERS...

  7. 33 CFR 104.106 - Passenger access area.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... 33 Navigation and Navigable Waters 1 2011-07-01 2011-07-01 false Passenger access area. 104.106... SECURITY MARITIME SECURITY: VESSELS General § 104.106 Passenger access area. (a) A ferry, passenger vessel, or cruise ship may designate areas within the vessel as passenger access areas. (b) A...

  8. 46 CFR 72.25-10 - Location of passenger quarters.

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ... 46 Shipping 3 2011-10-01 2011-10-01 false Location of passenger quarters. 72.25-10 Section 72.25-10 Shipping COAST GUARD, DEPARTMENT OF HOMELAND SECURITY (CONTINUED) PASSENGER VESSELS CONSTRUCTION AND ARRANGEMENT Passenger Accommodations § 72.25-10 Location of passenger quarters. (a) The...

  9. 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...

  10. 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...

  11. 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...

  12. 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...

  13. 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...

  14. 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.

  15. 46 CFR 171.045 - Weight of passengers and crew.

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ... 46 Shipping 7 2011-10-01 2011-10-01 false Weight of passengers and crew. 171.045 Section 171.045... PERTAINING TO VESSELS CARRYING PASSENGERS Intact Stability § 171.045 Weight of passengers and crew. (a) This... requirements applicable to each vessel, using a total weight of passengers and crew carried, is based upon...

  16. 46 CFR 122.515 - Passenger safety bill.

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ... accommodations for more than 49 passengers. (b) Each passenger safety bill required by this section must list: (1... 46 Shipping 4 2010-10-01 2010-10-01 false Passenger safety bill. 122.515 Section 122.515 Shipping COAST GUARD, DEPARTMENT OF HOMELAND SECURITY (CONTINUED) SMALL PASSENGER VESSELS CARRYING MORE THAN...

  17. 19 CFR 122.88 - Aircraft carrying domestic (stopover) passengers.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 19 Customs Duties 1 2010-04-01 2010-04-01 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...

  18. Teaching Children To Read and Write: Becoming an Influential Teacher.

    ERIC Educational Resources Information Center

    Ruddell, Robert B.; Ruddell, Martha Rapp

    Designed to include a broad base of information about how children acquire and develop literacy, this book discusses the knowledge that is necessary to be an influential teacher of reading and writing. Each chapter in the book begins and ends with a "Double Entry Journal" (DEJ)--an interactive strategy designed to stimulate thinking and to draw…

  19. 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.

  20. 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.

  1. Community centrality for node's influential ranking in complex network

    NASA Astrophysics Data System (ADS)

    Cai, Biao; Tuo, Xian-Guo; Yang, Kai-Xue; Liu, Ming-Zhe

    2014-10-01

    Some tiny party of influential nodes may highly affect spread of information in complex networks. For the case of very high time complexity in the shortest path computation of global centralities, making use of local community centrality to identify influential nodes is an open and possible problem. Compared to degree and local centralities, a five-heartbeat forward community centrality is proposed in this paper, in which a five-step induced sub-graph of certain node in the network will be achieved. Next, we induce the minimal spanning tree (MMT) of the sub-graph. Finally, we take the sum of all weights of the MMT as community centrality measurement that needs to be the influential ranking of the node. We use the susceptible, infected and recovered (SIR) model to evaluate the performance of this method on several public test network data and explore the forward steps of community centrality by experiments. Simulative results show that our method with five steps can identify the influential ranking of nodes in complex network as well.

  2. Ten Lives in Mine: Creating Portraits of Influential Chinese Educators

    ERIC Educational Resources Information Center

    Hayhoe, Ruth

    2004-01-01

    This is an essay about research methodology, particularly the use of narrative method, in a project which depicts portraits of 10 influential educators of China. The essay begins with reflections on the dialogue of civilizations and the potential future influence of Chinese civilization in the global community. It then goes on to consider the…

  3. Optimal boarding method for airline passengers

    SciTech Connect

    Steffen, Jason H.; /Fermilab

    2008-02-01

    Using a Markov Chain Monte Carlo optimization algorithm and a computer simulation, I find the passenger ordering which minimizes the time required to board the passengers onto an airplane. The model that I employ assumes that the time that a passenger requires to load his or her luggage is the dominant contribution to the time needed to completely fill the aircraft. The optimal boarding strategy may reduce the time required to board and airplane by over a factor of four and possibly more depending upon the dimensions of the aircraft. I explore some features of the optimal boarding method and discuss practical modifications to the optimal. Finally, I mention some of the benefits that could come from implementing an improved passenger boarding scheme.

  4. Multiple Infections with Cardinium and Two Strains of Wolbachia in The Spider Mite Tetranychus phaselus Ehara: Revealing New Forces Driving the Spread of Wolbachia

    PubMed Central

    Zhao, Dong-Xiao; Chen, Da-Song; Ge, Cheng; Gotoh, Tetsuo; Hong, Xiao-Yue

    2013-01-01

    Cytoplasmic incompatibility (CI) has been proposed as a major mechanism by which certain strains of Wolbachia to invade and persist in host populations. However, mechanisms that underlie the invasion and persistence of non-CI strains are less well understood. Here, we established a spider mite Tetranychus phaselus population multiply infected by Cardinium as well as two distinct lineages of Wolbachia, designated wCon and wOri, to study the forces driving the spread of the non-CI strain of Wolbachia wOri. Interestingly, we found that wOri provided a longevity advantage to its female hosts under ideal conditions, making wOri stay longer in this population, and then being transmitted to more offspring. Furthermore, the lifespan of uninfected females was reduced when mated with multiple-infected males. As a result, the uninfected population is attenuated by the multiple-infected males. Thus, we infer that the host age effects of multiple infection may represent sufficient forces driving the spread of wOri through the host population. PMID:23355904

  5. Proinflammatory Cytokine Gene Expression by Murine Macrophages in Response to Brugia malayi Wolbachia Surface Protein

    PubMed Central

    Porksakorn, Chantima; Nuchprayoon, Surang; Park, Kiwon; Scott, Alan L.

    2007-01-01

    Wolbachia, an endosymbiotic bacterium found in most species of filarial parasites, is thought to play a significant role in inducing innate inflammatory responses in lymphatic filariasis patients. However, the Wolbachia-derived molecules that are recognized by the innate immune system have not yet been identified. In this study, we exposed the murine macrophage cell line RAW 264.7 to a recombinant form of the major Wolbachia surface protein (rWSP) to determine if WSP is capable of innately inducing cytokine transcription. Interleukin (IL)-1β, IL-6, and tumor necrosis factor (TNF) mRNAs were all upregulated by the rWSP stimulation in a dose-dependant manner. TNF transcription peaked at 3 hours, whereas IL-1β and IL-6 transcription peaked at 6 hours post-rWSP exposure. The levels of innate cytokine expression induced by a high-dose (9.0 μg/mL) rWSP in the RAW 264.7 cells were comparable to the levels induced by 0.1 μg/mL E. coli-derived lipopolysaccharides. Pretreatment of the rWSP with proteinase-K drastically reduced IL-1β, IL-6, and TNF transcription. However, the proinflammatory response was not inhibited by polymyxin B treatment. These results strongly suggest that the major Wolbachia surface protein molecule WSP is an important inducer of innate immune responses during filarial infections. PMID:17641731

  6. 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

  7. 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. PMID:25948567

  8. Targeting the Wolbachia Cell Division Protein FtsZ as a New Approach for Antifilarial Therapy

    PubMed Central

    Li, Zhiru; Garner, Amanda L.; Gloeckner, Christian; Janda, Kim D.; Carlow, Clotilde K.

    2011-01-01

    The use of antibiotics targeting the obligate bacterial endosymbiont Wolbachia of filarial parasites has been validated as an approach for controlling filarial infection in animals and humans. Availability of genomic sequences for the Wolbachia (wBm) present in the human filarial parasite Brugia malayi has enabled genome-wide searching for new potential drug targets. In the present study, we investigated the cell division machinery of wBm and determined that it possesses the essential cell division gene ftsZ which was expressed in all developmental stages of B. malayi examined. FtsZ is a GTPase thereby making the protein an attractive Wolbachia drug target. We described the molecular characterization and catalytic properties of Wolbachia FtsZ. We also demonstrated that the GTPase activity was inhibited by the natural product, berberine, and small molecule inhibitors identified from a high-throughput screen. Furthermore, berberine was also effective in reducing motility and reproduction in B. malayi parasites in vitro. Our results should facilitate the discovery of selective inhibitors of FtsZ as a novel anti-symbiotic approach for controlling filarial infection. Note The nucleotide sequences reported in this paper are available in GenBank™ Data Bank under the accession number wAlB-FtsZ (JN616286). PMID:22140592

  9. 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. PMID:26470291

  10. Models to assess how best to replace dengue virus vectors with Wolbachia-infected mosquito populations.

    PubMed

    Zhang, Xianghong; Tang, Sanyi; Cheke, Robert A

    2015-11-01

    Dengue fever is increasing in importance in the tropics and subtropics. Endosymbiotic Wolbachia bacteria as novel control methods can reduce the ability of virus transmission. So, many mosquitoes infected with Wolbachia are released in some countries so that strategies for population replacement can be fulfilled. However, not all of these field trails are successful, for example, releases on Tri Nguyen Island, Vietnam in 2013 failed. Thus, we evaluated a series of relevant issues such as (a) why do some releases fail? (b) What affects the success of population replacement? And (c) Whether or not augmentation can block the dengue diseases in field trials. If not, how we can success be achieved? Models with and without augmentation, incorporating the effects of cytoplasmic incompatibility (CI) and fitness effects are proposed to describe the spread of Wolbachia in mosquito populations. Stability analysis revealed that backward bifurcations and multiple attractors may exist, which indicate that initial quantities of infected and uninfected mosquitoes, augmentation methods (timing, quantity, order and frequency) may affect the success of the strategies. The results show that successful population replacement will rely on selection of suitable strains of Wolbachia and careful design of augmentation methods. PMID:26407645

  11. 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

  12. 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...

  13. 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...

  14. Influence of the Virus LbFV and of Wolbachia in a Host-Parasitoid Interaction

    PubMed Central

    Woolfit, Megan; Vavre, Fabrice; O'Neill, Scott L.; Varaldi, Julien

    2012-01-01

    Symbionts are widespread and might have a substantial effect on the outcome of interactions between species, such as in host-parasitoid systems. Here, we studied the effects of symbionts on the outcome of host-parasitoid interactions in a four-partner system, consisting of the parasitoid wasp Leptopilina boulardi, its two hosts Drosophila melanogaster and D. simulans, the wasp virus LbFV, and the endosymbiotic bacterium Wolbachia. The virus is known to manipulate the superparasitism behavior of the parasitoid whereas some Wolbachia strains can reproductively manipulate and/or confer pathogen protection to Drosophila hosts. We used two nuclear backgrounds for both Drosophila species, infected with or cured of their respective Wolbachia strains, and offered them to L. boulardi of one nuclear background, either infected or uninfected by the virus. The main defence mechanism against parasitoids, i.e. encapsulation, and other important traits of the interaction were measured. The results showed that virus-infected parasitoids are less frequently encapsulated than uninfected ones. Further experiments showed that this viral effect involved both a direct protective effect against encapsulation and an indirect effect of superparasitism. Additionally, the Wolbachia strain wAu affected the encapsulation ability of its Drosophila host but the direction of this effect was strongly dependent on the presence/absence of LbFV. Our results confirmed the importance of heritable symbionts in the outcome of antagonistic interactions. PMID:22558118

  15. The diversity of reproductive parasites among arthropods: Wolbachia do not walk alone

    PubMed Central

    Duron, Olivier; Bouchon, Didier; Boutin, Sébastien; Bellamy, Lawrence; Zhou, Liqin; Engelstädter, Jan; Hurst, Gregory D

    2008-01-01

    Background Inherited bacteria have come to be recognised as important components of arthropod biology. In addition to mutualistic symbioses, a range of other inherited bacteria are known to act either as reproductive parasites or as secondary symbionts. Whilst the incidence of the α-proteobacterium Wolbachia is relatively well established, the current knowledge of other inherited bacteria is much weaker. Here, we tested 136 arthropod species for a range of inherited bacteria known to demonstrate reproductive parasitism, sampling each species more intensively than in past surveys. Results The inclusion of inherited bacteria other than Wolbachia increased the number of infections recorded in our sample from 33 to 57, and the proportion of species infected from 22.8% to 32.4%. Thus, whilst Wolbachia remained the dominant inherited bacterium, it alone was responsible for around half of all inherited infections of the bacteria sampled, with members of the Cardinium, Arsenophonus and Spiroplasma ixodetis clades each occurring in 4% to 7% of all species. The observation that infection was sometimes rare within host populations, and that there was variation in presence of symbionts between populations indicates that our survey will itself underscore incidence. Conclusion This extensive survey demonstrates that at least a third of arthropod species are infected by a diverse assemblage of maternally inherited bacteria that are likely to strongly influence their hosts' biology, and indicates an urgent need to establish the nature of the interaction between non-Wolbachia bacteria and their hosts. PMID:18577218

  16. Dynamics of the "popcorn" Wolbachia infection in outbred Aedes aegypti informs prospects for mosquito vector control.

    PubMed

    Yeap, H L; Mee, P; Walker, T; Weeks, A R; O'Neill, S L; Johnson, P; Ritchie, S A; Richardson, K M; Doig, C; Endersby, N M; Hoffmann, A A

    2011-02-01

    Forty percent of the world's population is at risk of contracting dengue virus, which produces dengue fever with a potentially fatal hemorrhagic form. The wMelPop Wolbachia infection of Drosophila melanogaster reduces life span and interferes with viral transmission when introduced into the mosquito Aedes aegypti, the primary vector of dengue virus. Wolbachia has been proposed as an agent for preventing transmission of dengue virus. Population invasion by Wolbachia depends on levels of cytoplasmic incompatibility, fitness effects, and maternal transmission. Here we characterized these traits in an outbred genetic background of a potential target population of Ae. aegypti using two crossing schemes. Cytoplasmic incompatibility was strong in this background, and the maternal transmission rate of Wolbachia was high. The infection substantially reduced longevity of infected adult females, regardless of whether adults came from larvae cultured under high or low levels of nutrition or density. The infection reduced the viability of diapausing and nondiapausing eggs. Viability was particularly low when eggs were laid by older females and when diapausing eggs had been stored for a few weeks. The infection affected mosquito larval development time and adult body size under different larval nutrition levels and densities. The results were used to assess the potential for wMelPop-CLA to invade natural populations of Ae. aegypti and to develop recommendations for the maintenance of fitness in infected mosquitoes that need to compete against field insects. PMID:21135075

  17. 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...

  18. 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. PMID:23802459

  19. 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

  20. Composition of bacterial communities associated with natural and laboratory populations of Asobara tabida infected with Wolbachia.

    PubMed

    Zouache, Karima; Voronin, Denis; Tran-Van, Van; Mavingui, Patrick

    2009-06-01

    Asobara tabida wasps are fly endoparasitoids that naturally harbor three Wolbachia strains, which induce cytoplasmic incompatibility and control oogenesis. To investigate whether other bacteria play a role in wasp biology, we surveyed the bacterial communities of wild A. tabida populations originating from different regions of France and of laboratory colonies using PCR-denaturing gradient gel electrophoresis and culture methods. Proteobacteria and Firmicutes were found to be the main phyla represented in these populations. Among these were several cultured and uncultured representatives of the genera Acetobacter, Acidomonas, Bacillus, Brevibacillus, Duganella, Herbaspirillum, Pseudomonas, Staphylococcus, and Streptococcus. In addition to Wolbachia, wild individuals harbored Rickettsia, which tended to be lost when insects were reared in the laboratory. The antibiotic treatment used to generate wasp sublines singly infected with Wolbachia also affected the overall bacterial composition, with most fingerprint sequences being characteristic of the family Enterobacteriaceae. We also screened for potentially heritable endosymbionts by PCR and fluorescence in situ hybridization in stable laboratory lines, with only Wolbachia being consistently found in wasp ovaries. PMID:19376923

  1. 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.

  2. Deleterious Passengers in Adapting Populations

    PubMed Central

    Good, Benjamin H.; Desai, Michael M.

    2014-01-01

    Most new mutations are deleterious and are eventually eliminated by natural selection. But in an adapting population, the rapid amplification of beneficial mutations can hinder the removal of deleterious variants in nearby regions of the genome, altering the patterns of sequence evolution. Here, we analyze the interactions between beneficial “driver” mutations and linked deleterious “passengers” during the course of adaptation. We derive analytical expressions for the substitution rate of a deleterious mutation as a function of its fitness cost, as well as the reduction in the beneficial substitution rate due to the genetic load of the passengers. We find that the fate of each deleterious mutation varies dramatically with the rate and spectrum of beneficial mutations and the deleterious substitution rate depends nonmonotonically on the population size and the rate of adaptation. By quantifying this dependence, our results allow us to estimate which deleterious mutations will be likely to fix and how many of these mutations must arise before the progress of adaptation is significantly reduced. PMID:25194161

  3. 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

  4. Potential involvement of Brugia malayi cysteine proteases in the maintenance of the endosymbiotic relationship with Wolbachia

    PubMed Central

    Lustigman, Sara; Melnikow, Elena; Anand, Setty Balakrishnan; Contreras, Aroha; Nandi, Vijay; Liu, Jing; Bell, Aaron; Unnasch, Thomas R.; Rogers, Mathew B.; Ghedin, Elodie

    2014-01-01

    Brugia malayi, a parasitic nematode that causes lymphatic filariasis, harbors endosymbiotic intracellular bacteria, Wolbachia, that are required for the development and reproduction of the worm. The essential nature of this endosymbiosis led to the development of anti-Wolbachia chemotherapeutic approaches for the treatment of human filarial infections. Our study is aimed at identifying specific proteins that play a critical role in this endosymbiotic relationship leading to the identification of potential targets in the adult worms. Filarial cysteine proteases are known to be involved in molting and embryogenesis, processes shown to also be Wolbachia dependent. Based on the observation that cysteine protease transcripts are differentially regulated in response to tetracycline treatment, we focused on defining their role in symbiosis. We observe a bimodal regulation pattern of transcripts encoding cysteine proteases when in vitro tetracycline treated worms were examined. Using tetracycline-treated infertile female worms and purified embryos we established that the first peak of the bimodal pattern corresponds to embryonic transcripts while the second takes place within the hypodermis of the adult worms. Localization studies of the native proteins corresponding to Bm-cpl-3 and Bm-cpl-6 indicate that they are present in the area surrounding Wolbachia, and, in some cases, the proteins appear localized within the bacteria. Both proteins were also found in the inner bodies of microfilariae. The possible role of these cysteine proteases during development and endosymbiosis was further characterized using RNAi. Reduction in Bm-cpl-3 and Bm-cpl-6 transcript levels was accompanied by hindered microfilarial development and release, and reduced Wolbachia DNA levels, making these enzymes strong drug target candidates. PMID:25516837

  5. Using Wolbachia-based release for suppression of Aedes mosquitoes: insights from genetic data and population simulations.

    PubMed

    Rasić, Gordana; Endersby, Nancy M; Williams, Craig; Hoffmann, Ary A

    2014-07-01

    A novel strategy for suppressing disease transmission by Aedes aegypti, the main vector of dengue, uses releases of mosquitoes infected with the bacterium Wolbachia pipientis. Wolbachia are currently released to interfere with viral transmission, but there is also potential to use strains in mosquito suppression and elimination programs via the deleterious effects of the bacterium on the host. Mosquito suppression depends on target areas being relatively isolated to prevent reinvasion and on local climatic conditions. Here we explored the opportunity for suppression of A. aegypti in central Queensland, Australia, by using microsatellite data and simulations based on CIMSiM models of local weather conditions and breeding container data. Our results indicate that Wolbachia-induced extinctions in central Queensland are possible, although they may eventually be compromised by ongoing mosquito migration between towns until these sources are also suppressed. The results highlight a novel use of deleterious Wolbachia infections to achieve ecological as well as disease-related endpoints. PMID:25154109

  6. 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

  7. 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. PMID:24345405

  8. Absence of the Filarial Endosymbiont Wolbachia in Seal Heartworm (Acanthocheilonema spirocauda) but Evidence of Ancient Lateral Gene Transfer.

    PubMed

    Keroack, Caroline D; Wurster, Jenna I; Decker, Caroline G; Williams, Kalani M; Slatko, Barton E; Foster, Jeremy M; Williams, Steven A

    2016-06-01

    The symbiotic relationship of Wolbachia spp. was first observed in insects and subsequently in many parasitic filarial nematodes. This bacterium is believed to provide metabolic and developmental assistance to filarial parasitic nematodes, although the exact nature of this relationship remains to be fully elucidated. While Wolbachia is present in most filarial nematodes in the family Onchocercidae, it is absent in several disparate species such as the human parasite Loa loa . All tested members of the genus Acanthocheilonema, such as Acanthocheilonema viteae, have been shown to lack Wolbachia. Consistent with this, we show that Wolbachia is absent from the seal heartworm (Acanthocheilonema spirocauda), but lateral gene transfer (LGT) of DNA sequences between Wolbachia and A. spirocauda has occurred, indicating a past evolutionary association. Seal heartworm is an important pathogen of phocid seals and understanding its basic biology is essential for conservation of the host. The findings presented here may allow for the development of future treatments or diagnostics for the disease and also aid in clarification of the complicated nematode-Wolbachia relationship. PMID:26859724

  9. The effect of Wolbachia-induced cytoplasmic incompatibility on host population size in natural and manipulated systems.

    PubMed

    Dobson, Stephen L; Fox, Charles W; Jiggins, Francis M

    2002-03-01

    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

  10. 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

  11. 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

  12. 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.

  13. 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.

  14. 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. PMID:27415287

  15. 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. PMID:12433241

  16. 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

    2016-01-01

    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

  17. 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

  18. Construction of bacterial artificial chromosome libraries from the parasitic nematode Brugia malayi and physical mapping of the genome of its Wolbachia endosymbiont.

    PubMed

    Foster, Jeremy M; Kumar, Sanjay; Ganatra, Mehul B; Kamal, Ibrahim H; Ware, Jennifer; Ingram, Jessica; Pope-Chappell, Jesse; Guiliano, David; Whitton, Claire; Daub, Jennifer; Blaxter, Mark L; Slatko, Barton E

    2004-05-01

    The parasitic nematode, Brugia malayi, causes lymphatic filariasis in humans, which in severe cases leads to the condition known as elephantiasis. The parasite contains an endosymbiotic alpha-proteobacterium of the genus Wolbachia that is required for normal worm development and fecundity and is also implicated in the pathology associated with infections by these filarial nematodes. Bacterial artificial chromosome libraries were constructed from B. malayi DNA and provide over 11-fold coverage of the nematode genome. Wolbachia genomic fragments were simultaneously cloned into the libraries giving over 5-fold coverage of the 1.1 Mb bacterial genome. A physical framework for the Wolbachia genome was developed by construction of a plasmid library enriched for Wolbachia DNA as a source of sequences to hybridise to high-density bacterial artificial chromosome colony filters. Bacterial artificial chromosome end sequencing provided additional Wolbachia probe sequences to facilitate assembly of a contig that spanned the entire genome. The Wolbachia sequences provided a marker approximately every 10 kb. Four rare-cutting restriction endonucleases were used to restriction map the genome to a resolution of approximately 60 kb and demonstrate concordance between the bacterial artificial chromosome clones and native Wolbachia genomic DNA. Comparison of Wolbachia sequences to public databases using BLAST algorithms under stringent conditions allowed confident prediction of 69 Wolbachia peptide functions and two rRNA genes. Comparison to closely related complete genomes revealed that while most sequences had orthologs in the genome of the Wolbachia endosymbiont from Drosophila melanogaster, there was no evidence for long-range synteny. Rather, there were a few cases of short-range conservation of gene order extending over regions of less than 10 kb. The molecular scaffold produced for the genome of the Wolbachia from B. malayi forms the basis of a genomic sequencing effort for

  19. 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.

  20. 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

  1. 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

  2. 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

  3. 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

  4. 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. PMID:23349831

  5. Wolbachia infection suppresses both host defence and parasitoid counter-defence.

    PubMed

    Fytrou, Anastasia; Schofield, Peter G; Kraaijeveld, Alex R; Hubbard, Stephen F

    2006-04-01

    Endosymbiotic bacteria in the genus Wolbachia have been linked to several types of reproductive parasitism, which enhance their own transmission, while their direct effects on the host vary from beneficial to neutral or detrimental. Here, we report negative effects of infection on immunity-related traits of Drosophila simulans and the parasitoid wasp Leptopilina heterotoma. Infected D. simulans showed a reduced ability to encapsulate parasitoid eggs, compared to a tetracycline-treated, bacterium-free line. Challenging the two lines with a fungal pathogen, Beauveria bassiana, on the other hand, revealed no differences in survival. Moreover, elimination of Wolbachia was beneficial for the parasitoid wasp, as eggs laid by uninfected females suffered significantly lower encapsulation rates. We discuss possible origins of these fitness costs and their implications for infection dynamics and the interactions between host species. PMID:16618671

  6. 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

  7. 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

  8. Genetic Structure and Wolbachia Genotyping in Naturally Occurring Populations of Aedes albopictus across Contiguous Landscapes of Orissa, India

    PubMed Central

    Das, Biswadeep; Satapathy, Truptimayee; Kar, Santanu K.; Hazra, Rupenangshu K.

    2014-01-01

    Background Aedes albopictus has recently been implicated as a major vector in the emergence of dengue and chikungunya in several parts of India, like Orissa, which is gradually gaining endemicity for arboviral diseases. Ae. albopictus is further known to be naturally infected with Wolbachia (maternally inherited bacterium), which causes cytoplasmic incompatibility (CI) in mosquitoes leading to sperm-egg incompatibility inducing the death of embryo. Knowledge of genetic diversity of Ae. albopictus, along with revealing the type of Wolbachia infection in Ae. albopictus is important to explore the genetic and biological characteristics of Ae. albopictus, prior to exploring the uses of CI-based vector control strategies. In this study, we assessed the population genetic structure and the pattern of Wolbachia infection in Ae. albopictus mosquitoes of Orissa. Methods and Results Ae. albopictus mosquitoes were collected from 15 districts representing the four physiographical regions of Orissa from 2010–2012, analyzed for genetic variability at seven microsatellite loci and genotyped for Wolbachia strain detection using wsp gene primers. Most microsatellite markers were successfully amplified and were polymorphic, showing moderate genetic structure among all geographic populations (FST = 0.088). Genetic diversity was high (FST = 0.168) in Coastal Plains populations when compared with other populations, which was also evident from cluster analyses that showed most Coastal Plains populations consisted of a separate genetic cluster. Genotyping analyses revealed that Wolbachia-infected Ae. albopictus field populations of Orissa were mostly superinfected with wAlbA and wAlbB strains. Wolbachia superinfection was more pronounced in the Coastal Plain populations. Conclusion High genetic structure and Wolbachia superinfection, observed in the Coastal Plain populations of Orissa suggested it to be genetically and biologically more unique than other populations, and hence

  9. 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

  10. 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. PMID:26856916

  11. Wolbachia Variants Induce Differential Protection to Viruses in Drosophila melanogaster: A Phenotypic and Phylogenomic Analysis

    PubMed Central

    Chrostek, Ewa; Marialva, Marta S. P.; Esteves, Sara S.; Weinert, Lucy A.; Martinez, Julien; Jiggins, Francis M.; Teixeira, Luis

    2013-01-01

    Wolbachia are intracellular bacterial symbionts that are able to protect various insect hosts from viral infections. This tripartite interaction was initially described in Drosophila melanogaster carrying wMel, its natural Wolbachia strain. wMel has been shown to be genetically polymorphic and there has been a recent change in variant frequencies in natural populations. We have compared the antiviral protection conferred by different wMel variants, their titres and influence on host longevity, in a genetically identical D. melanogaster host. The phenotypes cluster the variants into two groups — wMelCS-like and wMel-like. wMelCS-like variants give stronger protection against Drosophila C virus and Flock House virus, reach higher titres and often shorten the host lifespan. We have sequenced and assembled the genomes of these Wolbachia, and shown that the two phenotypic groups are two monophyletic groups. We have also analysed a virulent and over-replicating variant, wMelPop, which protects D. melanogaster even better than the closely related wMelCS. We have found that a ∼21 kb region of the genome, encoding eight genes, is amplified seven times in wMelPop and may be the cause of its phenotypes. Our results indicate that the more protective wMelCS-like variants, which sometimes have a cost, were replaced by the less protective but more benign wMel-like variants. This has resulted in a recent reduction in virus resistance in D. melanogaster in natural populations worldwide. Our work helps to understand the natural variation in wMel and its evolutionary dynamics, and inform the use of Wolbachia in arthropod-borne disease control. PMID:24348259

  12. Within-species diversity of Wolbachia-induced cytoplasmic incompatibility in haplodiploid insects.

    PubMed

    Vavre, F; Dedeine, F; Quillon, M; Fouillet, P; Fleury, F; Bouletreau, M

    2001-08-01

    Wolbachia-induced cytoplasmic incompatibility (CI) can have two consequences in haplodiploid insects: fertilized eggs either die (female mortality, FM) or they develop into haploid males (male development, MD). Origin of this diversity remains poorly understood, but current hypotheses invoke variation in damage suffered by paternal chromosomes in incompatible eggs, thus intermediate CI types should be expected. Here, we show the existence of such a particular CI type. In the parasitoid wasp Leptopilina heterotoma, we compared CI effects in crosses involving lines derived from a single inbred line with various Wolbachia infection statuses (natural tri-infection, mono-infection, or no infection). Tri-infected males induce a FM CI type when crossed with either uninfected or mono-infected females. Crossing mono-infected males with uninfected females results in almost complete CI with both reduced offspring production, indicating partial mortality of fertilized eggs, and increased number of sons, showing haploid male development of others. Mono-infected males thus induce an intermediate Cl type when mated with uninfected females. The first evidence of this expected particular CI type demonstrates that no discontinuity separates MD and FM CI types, which appear to be end points of a phenotypic continuum. Second, different CI types can occur within a given species and even within offspring of a single pair. Third, phenotypic expression of the particular CI type induced by a given Wolbachia variant depends on other bacterial variants that co-infect the same tissues. These results support the idea that haplodiploids should be helpful in clarifying evolutionary pathways of insect-Wolbachia associations. PMID:11580031

  13. 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...

  14. 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...

  15. 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...

  16. 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...

  17. 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...

  18. 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...

  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, 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...

  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. 14 CFR 125.327 - 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. 125... § 125.327 Briefing of passengers before flight. (a) Before each takeoff, each pilot in command of an... briefing shall include a statement that the Federal Aviation Regulations require passenger compliance...

  4. 46 CFR 185.506 - Passenger safety orientation.

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ... 46 Shipping 7 2011-10-01 2011-10-01 false Passenger safety orientation. 185.506 Section 185.506 Shipping COAST GUARD, DEPARTMENT OF HOMELAND SECURITY (CONTINUED) SMALL PASSENGER VESSELS (UNDER 100 GROSS... safety orientation. If only a small number of passengers embark at a port after the original muster...

  5. 46 CFR 122.506 - Passenger safety orientation.

    Code of Federal Regulations, 2012 CFR

    2012-10-01

    ... 46 Shipping 4 2012-10-01 2012-10-01 false Passenger safety orientation. 122.506 Section 122.506 Shipping COAST GUARD, DEPARTMENT OF HOMELAND SECURITY (CONTINUED) SMALL PASSENGER VESSELS CARRYING MORE.... If only a small number of passengers embark at a port after the original muster has been held,...

  6. 46 CFR 185.502 - Crew and passenger list.

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ... 46 Shipping 7 2011-10-01 2011-10-01 false Crew and passenger list. 185.502 Section 185.502 Shipping COAST GUARD, DEPARTMENT OF HOMELAND SECURITY (CONTINUED) SMALL PASSENGER VESSELS (UNDER 100 GROSS TONS) OPERATIONS Preparations for Emergencies § 185.502 Crew and passenger list. (a) The...

  7. 46 CFR 122.506 - Passenger safety orientation.

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ... 46 Shipping 4 2011-10-01 2011-10-01 false Passenger safety orientation. 122.506 Section 122.506 Shipping COAST GUARD, DEPARTMENT OF HOMELAND SECURITY (CONTINUED) SMALL PASSENGER VESSELS CARRYING MORE.... If only a small number of passengers embark at a port after the original muster has been held,...

  8. 46 CFR 122.506 - Passenger safety orientation.

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ... 46 Shipping 4 2013-10-01 2013-10-01 false Passenger safety orientation. 122.506 Section 122.506 Shipping COAST GUARD, DEPARTMENT OF HOMELAND SECURITY (CONTINUED) SMALL PASSENGER VESSELS CARRYING MORE.... If only a small number of passengers embark at a port after the original muster has been held,...

  9. 46 CFR 185.506 - Passenger safety orientation.

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    ... 46 Shipping 7 2014-10-01 2014-10-01 false Passenger safety orientation. 185.506 Section 185.506 Shipping COAST GUARD, DEPARTMENT OF HOMELAND SECURITY (CONTINUED) SMALL PASSENGER VESSELS (UNDER 100 GROSS... safety orientation. If only a small number of passengers embark at a port after the original muster...

  10. 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 Shipping COAST GUARD, DEPARTMENT OF HOMELAND SECURITY (CONTINUED) SMALL PASSENGER VESSELS (UNDER 100 GROSS TONS) OPERATIONS Preparations for Emergencies § 185.502 Crew and passenger list. (a) The...

  11. 46 CFR 185.502 - Crew and passenger list.

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    ... 46 Shipping 7 2014-10-01 2014-10-01 false Crew and passenger list. 185.502 Section 185.502 Shipping COAST GUARD, DEPARTMENT OF HOMELAND SECURITY (CONTINUED) SMALL PASSENGER VESSELS (UNDER 100 GROSS TONS) OPERATIONS Preparations for Emergencies § 185.502 Crew and passenger list. (a) The...

  12. 46 CFR 185.506 - Passenger safety orientation.

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ... 46 Shipping 7 2013-10-01 2013-10-01 false Passenger safety orientation. 185.506 Section 185.506 Shipping COAST GUARD, DEPARTMENT OF HOMELAND SECURITY (CONTINUED) SMALL PASSENGER VESSELS (UNDER 100 GROSS... safety orientation. If only a small number of passengers embark at a port after the original muster...

  13. 46 CFR 185.506 - Passenger safety orientation.

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ... 46 Shipping 7 2010-10-01 2010-10-01 false Passenger safety orientation. 185.506 Section 185.506 Shipping COAST GUARD, DEPARTMENT OF HOMELAND SECURITY (CONTINUED) SMALL PASSENGER VESSELS (UNDER 100 GROSS... safety orientation. If only a small number of passengers embark at a port after the original muster...

  14. 46 CFR 185.502 - Crew and passenger list.

    Code of Federal Regulations, 2012 CFR

    2012-10-01

    ... 46 Shipping 7 2012-10-01 2012-10-01 false Crew and passenger list. 185.502 Section 185.502 Shipping COAST GUARD, DEPARTMENT OF HOMELAND SECURITY (CONTINUED) SMALL PASSENGER VESSELS (UNDER 100 GROSS TONS) OPERATIONS Preparations for Emergencies § 185.502 Crew and passenger list. (a) The...

  15. 46 CFR 185.502 - Crew and passenger list.

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ... 46 Shipping 7 2013-10-01 2013-10-01 false Crew and passenger list. 185.502 Section 185.502 Shipping COAST GUARD, DEPARTMENT OF HOMELAND SECURITY (CONTINUED) SMALL PASSENGER VESSELS (UNDER 100 GROSS TONS) OPERATIONS Preparations for Emergencies § 185.502 Crew and passenger list. (a) The...

  16. 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 Shipping COAST GUARD, DEPARTMENT OF HOMELAND SECURITY (CONTINUED) SMALL PASSENGER VESSELS CARRYING MORE.... If only a small number of passengers embark at a port after the original muster has been held,...

  17. 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 Shipping COAST GUARD, DEPARTMENT OF HOMELAND SECURITY (CONTINUED) SMALL PASSENGER VESSELS (UNDER 100 GROSS... safety orientation. If only a small number of passengers embark at a port after the original muster...

  18. 46 CFR 122.506 - Passenger safety orientation.

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ... 46 Shipping 4 2010-10-01 2010-10-01 false Passenger safety orientation. 122.506 Section 122.506 Shipping COAST GUARD, DEPARTMENT OF HOMELAND SECURITY (CONTINUED) SMALL PASSENGER VESSELS CARRYING MORE.... If only a small number of passengers embark at a port after the original muster has been held,...

  19. 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 Safety belts: passenger warning device. (a) If there are means to indicate to the passengers when safety belts should be fastened, they must be installed to be operated from either pilot seat. (b)...

  20. 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 Safety belts: passenger warning device. (a) If there are means to indicate to the passengers when safety belts should be fastened, they must be installed to be operated from either pilot seat. (b)...

  1. 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 Safety belts: passenger warning device. (a) If there are means to indicate to the passengers when safety belts should be fastened, they must be installed to be operated from either pilot seat. (b)...

  2. 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 Safety belts: passenger warning device. (a) If there are means to indicate to the passengers when safety belts should be fastened, they must be installed to be operated from either pilot seat. (b)...

  3. 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 Safety belts: passenger warning device. (a) If there are means to indicate to the passengers when safety belts should be fastened, they must be installed to be operated from either pilot seat. (b)...

  4. 49 CFR 239.103 - Passenger train emergency simulations.

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ... 49 Transportation 4 2011-10-01 2011-10-01 false Passenger train emergency simulations. 239.103....103 Passenger train emergency simulations. (a) General. Each railroad operating passenger train service shall conduct full-scale emergency simulations, in order to determine its capability to...

  5. 46 CFR 15.530 - Large passenger vessels.

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ... 46 Shipping 1 2010-10-01 2010-10-01 false Large passenger vessels. 15.530 Section 15.530 Shipping... Manning Requirements; Inspected Vessels § 15.530 Large passenger vessels. (a) The owner or operator of a U.S. flag large passenger vessel must ensure that any non-resident alien holding a Coast...

  6. 49 CFR 541.5 - Requirements for passenger motor vehicles.

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ... 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 to... paragraphs (a)(1) through (a)(18) inclusive, if the part is present on the passenger motor vehicle. In...

  7. 14 CFR 23.791 - Passenger information signs.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... 14 Aeronautics and Space 1 2011-01-01 2011-01-01 false Passenger information signs. 23.791 Section... Personnel and Cargo Accommodations § 23.791 Passenger information signs. For those airplanes in which the... separated from the passenger compartment, there must be at least one illuminated sign (using either...

  8. 14 CFR 23.791 - Passenger information signs.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... 14 Aeronautics and Space 1 2014-01-01 2014-01-01 false Passenger information signs. 23.791 Section... Personnel and Cargo Accommodations § 23.791 Passenger information signs. For those airplanes in which the... separated from the passenger compartment, there must be at least one illuminated sign (using either...

  9. 14 CFR 23.791 - Passenger information signs.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... 14 Aeronautics and Space 1 2013-01-01 2013-01-01 false Passenger information signs. 23.791 Section... Personnel and Cargo Accommodations § 23.791 Passenger information signs. For those airplanes in which the... separated from the passenger compartment, there must be at least one illuminated sign (using either...

  10. 14 CFR 23.791 - Passenger information signs.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... 14 Aeronautics and Space 1 2012-01-01 2012-01-01 false Passenger information signs. 23.791 Section... Personnel and Cargo Accommodations § 23.791 Passenger information signs. For those airplanes in which the... separated from the passenger compartment, there must be at least one illuminated sign (using either...

  11. 14 CFR 23.791 - Passenger information signs.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 14 Aeronautics and Space 1 2010-01-01 2010-01-01 false Passenger information signs. 23.791 Section... Personnel and Cargo Accommodations § 23.791 Passenger information signs. For those airplanes in which the... separated from the passenger compartment, there must be at least one illuminated sign (using either...

  12. 49 CFR 239.103 - Passenger train emergency simulations.

    Code of Federal Regulations, 2012 CFR

    2012-10-01

    ... 49 Transportation 4 2012-10-01 2012-10-01 false Passenger train emergency simulations. 239.103....103 Passenger train emergency simulations. (a) General. Each railroad operating passenger train service shall conduct full-scale emergency simulations, in order to determine its capability to...

  13. 14 CFR 135.113 - Passenger occupancy of pilot seat.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 14 Aeronautics and Space 3 2010-01-01 2010-01-01 false Passenger occupancy of pilot seat. 135.113... Operations § 135.113 Passenger occupancy of pilot seat. No certificate holder may operate an aircraft type certificated after October 15, 1971, that has a passenger seating configuration, excluding any pilot seat,...

  14. 14 CFR 135.113 - Passenger occupancy of pilot seat.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... 14 Aeronautics and Space 3 2011-01-01 2011-01-01 false Passenger occupancy of pilot seat. 135.113... Operations § 135.113 Passenger occupancy of pilot seat. No certificate holder may operate an aircraft type certificated after October 15, 1971, that has a passenger seating configuration, excluding any pilot seat,...

  15. 49 CFR 177.870 - Regulations for passenger carrying vehicles.

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    ... Register citations affecting § 177.870 see the List of CFR Sections Affected, which appears in the Finding... carried in the passenger-carrying space of any motor vehicle transporting passengers for hire. (d... be transported by passenger-carrying aircraft or rail car may be transported on a motor...

  16. 49 CFR 177.870 - Regulations for passenger carrying vehicles.

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ... Register citations affecting § 177.870 see the List of CFR Sections Affected, which appears in the Finding... carried in the passenger-carrying space of any motor vehicle transporting passengers for hire. (d... be transported by passenger-carrying aircraft or rail car may be transported on a motor...

  17. 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…

  18. 46 CFR 46.05-1 - Passenger vessel.

    Code of Federal Regulations, 2012 CFR

    2012-10-01

    ... 46 Shipping 2 2012-10-01 2012-10-01 false Passenger vessel. 46.05-1 Section 46.05-1 Shipping COAST... VESSELS Definitions Used in This Part § 46.05-1 Passenger vessel. (a) For the purpose of the regulations in this part, a vessel is a passenger vessel if: (1) Engaged on an international voyage by sea,...

  19. 46 CFR 46.05-1 - Passenger vessel.

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ... 46 Shipping 2 2010-10-01 2010-10-01 false Passenger vessel. 46.05-1 Section 46.05-1 Shipping COAST... VESSELS Definitions Used in This Part § 46.05-1 Passenger vessel. (a) For the purpose of the regulations in this part, a vessel is a passenger vessel if: (1) Engaged on an international voyage by sea,...

  20. 46 CFR 46.05-1 - Passenger vessel.

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ... 46 Shipping 2 2013-10-01 2013-10-01 false Passenger vessel. 46.05-1 Section 46.05-1 Shipping COAST... VESSELS Definitions Used in This Part § 46.05-1 Passenger vessel. (a) For the purpose of the regulations in this part, a vessel is a passenger vessel if: (1) Engaged on an international voyage by sea,...

  1. 46 CFR 46.05-1 - Passenger vessel.

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    ... 46 Shipping 2 2014-10-01 2014-10-01 false Passenger vessel. 46.05-1 Section 46.05-1 Shipping COAST... VESSELS Definitions Used in This Part § 46.05-1 Passenger vessel. (a) For the purpose of the regulations in this part, a vessel is a passenger vessel if: (1) Engaged on an international voyage by sea,...

  2. 46 CFR 46.05-1 - Passenger vessel.

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ... 46 Shipping 2 2011-10-01 2011-10-01 false Passenger vessel. 46.05-1 Section 46.05-1 Shipping COAST... VESSELS Definitions Used in This Part § 46.05-1 Passenger vessel. (a) For the purpose of the regulations in this part, a vessel is a passenger vessel if: (1) Engaged on an international voyage by sea,...

  3. 14 CFR 29.807 - Passenger emergency exits.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... a crash landing must be extremely remote. (d) Ditching emergency exits for passengers. If... 14 Aeronautics and Space 1 2011-01-01 2011-01-01 false Passenger emergency exits. 29.807 Section... Accommodations § 29.807 Passenger emergency exits. (a) Type. For the purpose of this part, the types of...

  4. 75 FR 68467 - Transportation for Individuals With Disabilities: Passenger Vessels

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-11-08

    ... and practices of passenger vessel operators (PVOs). The rule was issued on July 6, 2010 (75 FR 38878... 49 CFR Part 39 RIN 2105-AB87 Transportation for Individuals With Disabilities: Passenger Vessels... Disabilities Act (ADA) final rule to ensure nondiscrimination on the basis of disability by passenger...

  5. Identifying influential nodes in weighted networks based on evidence theory

    NASA Astrophysics Data System (ADS)

    Wei, Daijun; Deng, Xinyang; Zhang, Xiaoge; Deng, Yong; Mahadevan, Sankaran

    2013-05-01

    The design of an effective ranking method to identify influential nodes is an important problem in the study of complex networks. In this paper, a new centrality measure is proposed based on the Dempster-Shafer evidence theory. The proposed measure trades off between the degree and strength of every node in a weighted network. The influences of both the degree and the strength of each node are represented by basic probability assignment (BPA). The proposed centrality measure is determined by the combination of these BPAs. Numerical examples are used to illustrate the effectiveness of the proposed method.

  6. Introduction to the alpha-proteobacteria: Wolbachia and Bartonella, Rickettsia, Brucella, Ehrlichia, and Anaplasma.

    PubMed

    Bowman, Dwight D

    2011-11-01

    Wolbachia is an obligate intracellular endosymbiont and likely mutualist living within the heartworm Dirofilaria immitis and a number of other filarial nematodes in the family Onchocercidae. The bacterial infection is passed from worm to worm transovarially; the organisms are in ovarian cells, the developing microfilariae, and multiply and persist in all later developmental stages through the mosquito and into the next host. Besides being present in the ovaries of the adult worms, they also are present in large numbers within the hypodermal tissues of the nematode. It is now know that these bacteria that were first observed in heartworms more than 30 years ago are actually related to similar Wolbachia bacteria that are found in arthropods. Wolbachia is an alpha-proteobacteria, and this group includes a number of important arthropod-transmitted bacterial agents of dogs and cats: Rickettsia rickettsii, R. felis, Anaplasma platys, Ehrlichia canis, E. chaffeensis, and E. ewingii. Alpha-proteobacteria are also important as obligate intracellular mutualists in plants in which they are responsible for nitrogen fixation. Recent work on the treatment of heartworms in dogs with doxycycline stems from related work with the human filarial nematode Onchocerca volvulus that causes river blindness in people. PMID:22152604

  7. Tropical Drosophila pandora carry Wolbachia infections causing cytoplasmic incompatibility or male killing.

    PubMed

    Richardson, Kelly M; Schiffer, Michele; Griffin, Philippa C; Lee, Siu F; Hoffmann, Ary A

    2016-08-01

    Wolbachia infections have been described in several Drosophila species, but relatively few have been assessed for phenotypic effects. Cytoplasmic incompatibility (CI) is the most common phenotypic effect that has been detected, while some infections cause male killing or feminization, and many Wolbachia infections have few host effects. Here, we describe two new infections in a recently described species, Drosophila pandora, one of which causes near-complete CI and near-perfect maternal transmission (the "CI" strain). The other infection is a male killer (the "MK" strain), which we confirm by observing reinitiation of male production following tetracycline treatment. No incompatibility was detected in crosses between CI strain males and MK strain females, and rare MK males do not cause CI. Molecular analyses indicate that the CI and MK infections are distantly related and the CI infection is closely related to the wRi infection of Drosophila simulans. Two population surveys indicate that all individuals are infected with Wolbachia, but the MK infection is uncommon. Given patterns of incompatibility among the strains, the infection dynamics is expected to be governed by the relative fitness of the females, suggesting that the CI infection should have a higher fitness. This was evidenced by changes in infection frequencies and sex ratios in population cages initiated at different starting frequencies of the infections. PMID:27282489

  8. Drosophila melanogaster brain invasion: pathogenic Wolbachia in central nervous system of the fly.

    PubMed

    Strunov, Anton; Kiseleva, Elena

    2016-04-01

    The pathogenic Wolbachia strain wMelPop rapidly over-replicates in the brain, muscles, and retina of Drosophila melanogaster, causing severe tissue degeneration and premature death of the host. The unique features of this endosymbiont make it an excellent tool to be used for biological control of insects, pests, and vectors of human diseases. To follow the dynamics of bacterial morphology and titer in the nerve cells we used transmission electron microscopy of 3-d-old female brains. The neurons and glial cells from central brain of the fly had different Wolbachia titers ranging from single bacteria to large accumulations, tearing cell apart and invading extracellular space. The neuropile regions of the brain were free of wMelPop. Wolbachia tightly interacted with host cell organelles and underwent several morphological changes in nerve cells. Based on different morphological types of bacteria described we propose for the first time a scheme of wMelPop dynamics within the somatic tissue of the host. PMID:25394184

  9. An ancient horizontal gene transfer between mosquito and the endosymbiotic bacterium Wolbachia pipientis.

    PubMed

    Woolfit, Megan; Iturbe-Ormaetxe, Iñaki; McGraw, Elizabeth A; O'Neill, Scott L

    2009-02-01

    The extent and biological relevance of horizontal gene transfer (HGT) in eukaryotic evolution remain highly controversial. Recent studies have demonstrated frequent and large-scale HGT from endosymbiotic bacteria to their hosts, but the great majority of these transferred genes rapidly become nonfunctional in the recipient genome. Here, we investigate an ancient HGT between a host metazoan and an endosymbiotic bacterium, Wolbachia pipientis. The transferred gene has so far been found only in mosquitoes and Wolbachia. In mosquitoes, it is a member of a gene family encoding candidate receptors required for malaria sporozoite invasion of the mosquito salivary gland. The gene copy in Wolbachia has substantially diverged in sequence from the mosquito homolog, is evolving under purifying selection, and is expressed, suggesting that this gene is also functional in the bacterial genome. Several lines of evidence indicate that the gene may have been transferred from eukaryotic host to bacterial endosymbiont. Regardless of the direction of transfer, however, these results demonstrate that interdomain HGT may give rise to functional, persistent, and possibly evolutionarily significant new genes. PMID:18988686

  10. Limited Dengue Virus Replication in Field-Collected Aedes aegypti Mosquitoes Infected with Wolbachia

    PubMed Central

    Frentiu, Francesca D.; Zakir, Tasnim; Walker, Thomas; Popovici, Jean; Pyke, Alyssa T.; van den Hurk, Andrew; McGraw, Elizabeth A.; O'Neill, Scott L.

    2014-01-01

    Introduction Dengue is one of the most widespread mosquito-borne diseases in the world. The causative agent, dengue virus (DENV), is primarily transmitted by the mosquito Aedes aegypti, a species that has proved difficult to control using conventional methods. The discovery that A. aegypti transinfected with the wMel strain of Wolbachia showed limited DENV replication led to trial field releases of these mosquitoes in Cairns, Australia as a biocontrol strategy for the virus. Methodology/Principal Findings Field collected wMel mosquitoes that were challenged with three DENV serotypes displayed limited rates of body infection, viral replication and dissemination to the head compared to uninfected controls. Rates of dengue infection, replication and dissemination in field wMel mosquitoes were similar to those observed in the original transinfected wMel line that had been maintained in the laboratory. We found that wMel was distributed in similar body tissues in field mosquitoes as in laboratory ones, but, at seven days following blood-feeding, wMel densities increased to a greater extent in field mosquitoes. Conclusions/Significance Our results indicate that virus-blocking is likely to persist in Wolbachia-infected mosquitoes after their release and establishment in wild populations, suggesting that Wolbachia biocontrol may be a successful strategy for reducing dengue transmission in the field. PMID:24587459

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

    PubMed Central

    White, J A; Kelly, S E; Cockburn, S N; Perlman, S J; Hunter, M S

    2011-01-01

    Theory suggests that maternally inherited endosymbionts can promote their spread and persistence in host populations by enhancing the production of daughters by infected hosts, either by improving overall host fitness, or through reproductive manipulation. In the doubly infected parasitoid wasp Encarsia inaron, Wolbachia manipulates host reproduction through cytoplasmic incompatibility (CI), but Cardinium does not. We investigated the fitness costs and/or benefits of infection by each bacterium in differentially cured E. inaron as a potential explanation for persistence of Cardinium in this population. We introgressed lines infected with Wolbachia, Cardinium or both with the cured line to create a similar genetic background, and evaluated several parasitoid fitness parameters. We found that symbiont infection resulted in both fitness costs and benefits for E. inaron. The cost was lower initial egg load for all infected wasps. The benefit was increased survivorship, which in turn increased male production for wasps infected with only Cardinium. Female production was unaffected by symbiont infection; we therefore have not yet identified a causal fitness effect that can explain the persistence of Cardinium in the population. Interestingly, the Cardinium survivorship benefit was not evident when Wolbachia was also present in the host, and the reproduction of doubly infected individuals did not differ significantly from uninfected wasps. Therefore, the results of our study show that even when multiple infections seem to have no effect on a host, there may be a complex interaction of costs and benefits among symbionts. PMID:20606691

  12. Differential induction of Th2- and Th1-associated responses by filarial antigens and endosymbiotic Wolbachia in a murine model of river blindness.

    PubMed

    Gentil, K; Hoerauf, A; Pearlman, E

    2012-06-01

    Immune responses to filarial parasites like the river blindness inducing Onchocerca volvulus are obscured by combined reactions to the filarial nematodes themselves and their endosymbiont bacteria Wolbachia. Overall, infection with filarial nematodes induces a strong Th2 response characterized by IL-5 production and to a lesser degree a Th1 response and IFNγ production. Neutrophil and eosinophil infiltration into the corneal stroma are hallmark features of Onchocerca volvulus stimulation in a mouse model of river blindness. To determine the splenic and corneal response to filarial antigens in the absence of Wolbachia, C57BL/6 mice were immunized subcutaneously with either endosymbiotic Wolbachia alone, a soluble extract from the filaria Acanthocheilonema viteae that does not contain Wolbachia, or both, and injected into the corneal stroma. Neutrophil and eosinophil infiltration into the cornea was assessed by immunohistochemistry. In addition, Th1- and Th2-associated responses to filaria or Wolbachia were investigated by determining IL-5 and IFN-γ production by splenocytes. We found that A. viteae in the absence of Wolbachia induced IL-5 production and eosinophil infiltration, but not IFN-γ. Conversely, Wolbachia induced IFN-γ production and no migration of eosinophils. There was no difference in neutrophil infiltration. Together, these findings demonstrate a distinct Th-associated phenotype induced by filaria and Wolbachia. PMID:24672682

  13. Replacing a Native Wolbachia with a Novel Strain Results in an Increase in Endosymbiont Load and Resistance to Dengue Virus in a Mosquito Vector

    PubMed Central

    Lu, Peng; Xi, Zhiyong

    2013-01-01

    Wolbachia is a maternally transmitted endosymbiotic bacterium that is estimated to infect up to 65% of insect species. The ability of Wolbachia to both induce pathogen interference and spread into mosquito vector populations makes it possible to develop Wolbachia as a biological control agent for vector-borne disease control. Although Wolbachia induces resistance to dengue virus (DENV), filarial worms, and Plasmodium in mosquitoes, species like Aedes polynesiensis and Aedes albopictus, which carry native Wolbachia infections, are able to transmit dengue and filariasis. In a previous study, the native wPolA in Ae. polynesiensis was replaced with wAlbB from Ae. albopictus, and resulted in the generation of the transinfected “MTB” strain with low susceptibility for filarial worms. In this study, we compare the dynamics of DENV serotype 2 (DENV-2) within the wild type “APM” strain and the MTB strain of Ae. polynesiensis by measuring viral infection in the mosquito whole body, midgut, head, and saliva at different time points post infection. The results show that wAlbB can induce a strong resistance to DENV-2 in the MTB mosquito. Evidence also supports that this resistance is related to a dramatic increase in Wolbachia density in the MTB's somatic tissues, including the midgut and salivary gland. Our results suggests that replacement of a native Wolbachia with a novel infection could serve as a strategy for developing a Wolbachia-based approach to target naturally infected insects for vector-borne disease control. PMID:23755311

  14. 75 FR 36300 - Enhancing Airline Passenger Protections

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-06-25

    ... April 11, 2000 (65 FR 19477-78), or you may visit http://DocketsInfo.dot.gov . Docket: For access to the... Airline Passenger Protections (75 FR 32318), which, among other things, solicits comment, without... the current practice of not prescribing carrier practices concerning the serving of peanuts. (75...

  15. Passenger Vans: A Transportation Concern for Camps.

    ERIC Educational Resources Information Center

    Gentle, Don; Gordon, Harold; Schainman, Steve

    2002-01-01

    In April 2002, the National Transportation Safety Board reissued a cautionary warning about the high accident potential of 15-passenger vans; some states have prohibited their use for transporting children. Three camp directors discuss legal and cost considerations for camps, possible alternative transportation, additional staff training needs,…

  16. Using Cooperatives to Transport Rural Passengers.

    ERIC Educational Resources Information Center

    Stommes, Eileen S.; Byrne, Robert J.

    A study of two rural New York counties--Cortland and Otsego--was undertaken to identify innovative ways to provide public passenger transportation services in rural areas by using transportation cooperatives. Information about current transportation services was obtained from staff members of the human service agencies and county government…

  17. 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...

  18. 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.

  19. 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) AIR TRAFFIC AND GENERAL OPERATING RULES GENERAL OPERATING AND FLIGHT RULES Fractional Ownership Operations Program Management § 91.1035...

  20. A Genetic Test of the Role of the Maternal Pronucleus in Wolbachia-Induced Cytoplasmic Incompatibility in Drosophila melanogaster

    PubMed Central

    Ferree, Patrick M.; Sullivan, William

    2006-01-01

    Cytoplasmic incompatibility (CI) is a reproductive sterility found in arthropods that is caused by the endoparasitic bacteria Wolbachia. In CI, host progeny fail to develop during early embryogenesis if Wolbachia-infected males fertilize uninfected females. It is widely accepted that this lethality is caused by some unknown Wolbachia-induced modification of the paternal nuclear material in the host testes. However, the direct means by which this modification leads to early embryonic death are currently unresolved. Results from previous studies suggested that CI lethality occurs as a result of asynchrony in cell cycle timing between the paternal and maternal pronuclei. This hypothesis can be tested experimentally by the prediction that the Wolbachia-modified paternal pronucleus should support androgenetic development (i.e., from the paternal pronucleus only). Using specific mutations in Drosophila melanogaster that produce androgenetic progeny, we demonstrate that the Wolbachia-induced modification inhibits this type of development. This result suggests that CI occurs independently of the maternal pronucleus and argues against pronuclear asynchrony as the primary cause of CI lethality. We propose that CI occurs instead as the result of either a developmentally incompetent paternal pronucleus or asynchrony between the paternal pronucleus and the cell cycle of the egg cytoplasm. PMID:16624919

  1. Identification and phylogenetic analysis of Dirofilaria ursi (Nematoda: Filarioidea) from Wisconsin black bears (Ursus americanus) and its Wolbachia endosymbiont.

    PubMed

    Michalski, Michelle L; Bain, Odile; Fischer, Kerstin; Fischer, Peter U; Kumar, Sanjay; Foster, Jeremy M

    2010-04-01

    Dirofilaria ursi is a filarial nematode of American black bears (Ursus americanus Pallas, 1780) that is vectored by black flies (Simuliidae) in many parts of the United States. In northwestern Wisconsin, the prevalence of microfilaremic bears during the fall hunting season was 21% (n = 47). Unsheathed blood microfilariae from Wisconsin bears possess characters consistent with the original description of D. ursi, as do adult worms observed histologically and grossly. Immunohistochemistry was used to identify the Wolbachia endosymbiont in the hypodermis and lateral cords of an adult female D. ursi. Amplification of wsp, gatB, coxA, fbpA, and ftsZ bacterial sequences from parasite DNA confirmed the presence of Wolbachia, and molecular phylogenetic analysis of the Wolbachia ftsZ gene groups the endosymbiont with Wolbachia from D. immitis and D. repens. Phylogenetic analysis of D. ursi 5s rDNA sequence confirms the morphological observations grouping this parasite as a member of Dirofilaria, and within the Dirofilaria - Onchocerca clade of filarial nematodes. This is the first report of Wolbachia characterization and molecular phylogeny information for D. ursi. PMID:19916630

  2. The "Genetic Program": Behind the Genesis of an Influential Metaphor.

    PubMed

    Peluffo, Alexandre E

    2015-07-01

    The metaphor of the "genetic program," indicating the genome as a set of instructions required to build a phenotype, has been very influential in biology despite various criticisms over the years. This metaphor, first published in 1961, is thought to have been invented independently in two different articles, one by Ernst Mayr and the other by François Jacob and Jacques Monod. Here, after a detailed analysis of what both parties meant by "genetic program," I show, using unpublished archives, the strong resemblance between the ideas of Mayr and Monod and suggest that their idea of genetic program probably shares a common origin. I explore the possibility that the two men met before 1961 and also exchanged their ideas through common friends and colleagues in the field of molecular biology. Based on unpublished correspondence of Jacob and Monod, I highlight the important events that influenced the preparation of their influential paper, which introduced the concept of the genetic program. Finally, I suggest that the genetic program metaphor may have preceded both papers and that it was probably used informally before 1961. PMID:26170444

  3. 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.

  4. 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. PMID:22181221

  5. 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.

  6. The Potential Use of Wolbachia-Based Mosquito Biocontrol Strategies for Japanese Encephalitis.

    PubMed

    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. The successful

  7. 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

  8. A Fast Estimation Method of Railway Passengers' Flow

    NASA Astrophysics Data System (ADS)

    Nagasaki, Yusaku; Asuka, Masashi; Komaya, Kiyotoshi

    To evaluate a train schedule from the viewpoint of passengers' convenience, it is important to know each passenger's choice of trains and transfer stations to arrive at his/her destination. Because of difficulties of measuring such passengers' behavior, estimation methods of railway passengers' flow are proposed to execute such an evaluation. However, a train schedule planning system equipped with those methods is not practical due to necessity of much time to complete the estimation. In this article, the authors propose a fast passengers' flow estimation method that employs features of passengers' flow graph using preparative search based on each train's arrival time at each station. And the authors show the results of passengers' flow estimation applied on a railway in an urban area.

  9. Detection of Wolbachia in the tick Ixodes ricinus is due to the presence of the hymenoptera endoparasitoid Ixodiphagus hookeri.

    PubMed

    Plantard, Olivier; Bouju-Albert, Agnès; Malard, Marie-Astrid; Hermouet, Axelle; Capron, Gilles; Verheyden, Hélène

    2012-01-01

    The identification of micro-organisms carried by ticks is an important issue for human and animal health. In addition to their role as pathogen vectors, ticks are also the hosts for symbiotic bacteria whose impact on tick biology is poorly known. Among these, the bacterium Wolbachia pipientis has already been reported associated with Ixodes ricinus and other tick species. However, the origins of Wolbachia in ticks and their consequences on tick biology (known to be very diverse in invertebrates, ranging from nutritional symbionts in nematodes to reproductive manipulators in insects) are unknown. Here we report that the endoparasitoid wasp Ixodiphagus hookeri (Hymenoptera, Chalcidoidea, Encyrtidae)--strictly associated with ticks for their development--infested at almost 100% prevalence by a W. pipientis strain belonging to a Wolbachia supergroup that has already been reported as associated with other hymenopteran parasitoids. In a natural population of I. ricinus that suffers high parasitism rates due to I. hookeri, we used specific PCR primers for both hymenopteran and W. pipientis gene fragments to show that all unfed tick nymphs parasitized by I. hookeri also harbored Wolbachia, while unparasitized ticks were Wolbachia-free. We demonstrated experimentally that unfed nymphs obtained from larvae exposed to I. hookeri while gorging on their vertebrate host also harbor Wolbachia. We hypothesize that previous studies that have reported W. pipientis in ticks are due to the cryptic presence of the endoparasitoid wasp I. hookeri. This association has remained hidden until now because parasitoids within ticks cannot be detected until engorgement of the nymphs brings the wasp eggs out of diapause. Finally, we discuss the consequences of this finding for our understanding of the tick microbiome, and their possible role in horizontal gene transfer among pathogenic and symbiotic bacteria. PMID:22292021

  10. Recent genome reduction of Wolbachia in Drosophila recens targets phage WO and narrows candidates for reproductive parasitism

    PubMed Central

    Metcalf, Jason A.; Jo, Minhee; Bordenstein, Sarah R.; Jaenike, John

    2014-01-01

    Wolbachia are maternally transmitted endosymbionts that often alter their arthropod hosts’ biology to favor the success of infected females, and they may also serve as a speciation microbe driving reproductive isolation. Two of these host manipulations include killing males outright and reducing offspring survival when infected males mate with uninfected females, a phenomenon known as cytoplasmic incompatibility. Little is known about the mechanisms behind these phenotypes, but interestingly either effect can be caused by the same Wolbachia strain when infecting different hosts. For instance, wRec causes cytoplasmic incompatibility in its native host Drosophila recens and male killing in D. subquinaria. The discovery of prophage WO elements in most arthropod Wolbachia has generated the hypothesis that WO may encode genes involved in these reproductive manipulations. However, PCR screens for the WO minor capsid gene indicated that wRec lacks phage WO. Thus, wRec seemed to provide an example where phage WO is not needed for Wolbachia-induced reproductive manipulation. To enable investigation of the mechanism of phenotype switching in different host backgrounds, and to examine the unexpected absence of phage WO, we sequenced the genome of wRec. Analyses reveal that wRec diverged from wMel approximately 350,000 years ago, mainly by genome reduction in the phage regions. While it lost the minor capsid gene used in standard PCR screens for phage WO, it retained two regions encompassing 33 genes, several of which have previously been associated with reproductive parasitism. Thus, WO gene involvement in reproductive manipulation cannot be excluded and reliance on single gene PCR should not be used to rule out the presence of phage WO in Wolbachia. Additionally, the genome sequence for wRec will enable transcriptomic and proteomic studies that may help elucidate the Wolbachia mechanisms of altered reproductive manipulations associated with host switching, perhaps among

  11. Detection of Wolbachia in the Tick Ixodes ricinus is Due to the Presence of the Hymenoptera Endoparasitoid Ixodiphagus hookeri

    PubMed Central

    Plantard, Olivier; Bouju-Albert, Agnès; Malard, Marie-Astrid; Hermouet, Axelle; Capron, Gilles; Verheyden, Hélène

    2012-01-01

    The identification of micro-organisms carried by ticks is an important issue for human and animal health. In addition to their role as pathogen vectors, ticks are also the hosts for symbiotic bacteria whose impact on tick biology is poorly known. Among these, the bacterium Wolbachia pipientis has already been reported associated with Ixodes ricinus and other tick species. However, the origins of Wolbachia in ticks and their consequences on tick biology (known to be very diverse in invertebrates, ranging from nutritional symbionts in nematodes to reproductive manipulators in insects) are unknown. Here we report that the endoparasitoid wasp Ixodiphagus hookeri (Hymenoptera, Chalcidoidea, Encyrtidae) – strictly associated with ticks for their development - is infested at almost 100% prevalence by a W. pipientis strain belonging to a Wolbachia supergroup that has already been reported as associated with other hymenopteran parasitoids. In a natural population of I. ricinus that suffers high parasitism rates due to I. hookeri, we used specific PCR primers for both hymenopteran and W. pipientis gene fragments to show that all unfed tick nymphs parasitized by I. hookeri also harbored Wolbachia, while unparasitized ticks were Wolbachia-free. We demonstrated experimentally that unfed nymphs obtained from larvae exposed to I. hookeri while gorging on their vertebrate host also harbor Wolbachia. We hypothesize that previous studies that have reported W. pipientis in ticks are due to the cryptic presence of the endoparasitoid wasp I. hookeri. This association has remained hidden until now because parasitoids within ticks cannot be detected until engorgement of the nymphs brings the wasp eggs out of diapause. Finally, we discuss the consequences of this finding for our understanding of the tick microbiome, and their possible role in horizontal gene transfer among pathogenic and symbiotic bacteria. PMID:22292021

  12. 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.

  13. Considerations for Passenger Transport by Advanced Spaceplanes

    NASA Astrophysics Data System (ADS)

    Scott-Scott, J. L.; Harrison, M.; Woodrow, A. D.

    One of the strongest incentives to space industry development in the 21st century is likely to be space tourism. At present the greatest handicaps to the exploitation of this opportunity are the prohibitive cost of access to space (few tourists could afford the ticket) and safety (astronautics remains an inherently dangerous occupation). This paper addresses both of these issues. First, the characteristics required of a vehicle suitable for carrying passengers into space are discussed in the context of the SKYLON spaceplane concept. Attention is focused on the design of the cabin module. Then the needs of the passengers are considered both from the perspectives of safety (the unique environmental hazards), health, and comfort. The paper concludes with a review of the key issues.

  14. [Problems of hygienic safety of passenger transportation].

    PubMed

    Sergeev, E P; Nedomerkov, Iu N; Shterengarts, R Ia; Noarov, Iu A

    1995-01-01

    Research in the sphere of railway hygiene helped develop sanitary regulations for railway stations, centers for preparation of passenger cars for exploitation, train restaurants, etc. The authors demonstrate the advances in prophylactic disinfection of stations and cars, in assessment of the hygienic hazards of polymeric materials used in car making and of the microclimate in the cars, etc. Trends for investigation in railway transport hygiene are outlined. PMID:7590382

  15. 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.

  16. Requirements for the Crash Protection of Older Vehicle Passengers

    PubMed Central

    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. PMID:12941224

  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. The expression of one ankyrin pk2 allele of the WO prophage is correlated with the Wolbachia feminizing effect in isopods

    PubMed Central

    2012-01-01

    Background The maternally inherited α-Proteobacteria Wolbachia pipientis is an obligate endosymbiont of nematodes and arthropods, in which they induce a variety of reproductive alterations, including Cytoplasmic Incompatibility (CI) and feminization. The genome of the feminizing wVulC Wolbachia strain harboured by the isopod Armadillidium vulgare has been sequenced and is now at the final assembly step. It contains an unusually high number of ankyrin motif-containing genes, two of which are homologous to the phage-related pk1 and pk2 genes thought to contribute to the CI phenotype in Culex pipiens. These genes encode putative bacterial effectors mediating Wolbachia-host protein-protein interactions via their ankyrin motifs. Results To test whether these Wolbachia homologs are potentially involved in altering terrestrial isopod reproduction, we determined the distribution and expression of both pk1 and pk2 genes in the 3 Wolbachia strains that induce CI and in 5 inducing feminization of their isopod hosts. Aside from the genes being highly conserved, we found a substantial copy number variation among strains, and that is linked to prophage diversity. Transcriptional analyses revealed expression of one pk2 allele (pk2b2) only in the feminizing Wolbachia strains of isopods. Conclusions These results reveal the need to investigate the functions of Wolbachia ankyrin gene products, in particular those of Pk2, and their host targets with respect to host sex manipulation. PMID:22497736

  19. Analysis of gene expression from the Wolbachia genome of a filarial nematode supports both metabolic and defensive roles within the symbiosis.

    PubMed

    Darby, Alistair C; Armstrong, Stuart D; Bah, Germanus S; Kaur, Gaganjot; Hughes, Margaret A; Kay, Suzanne M; Koldkjær, Pia; Rainbow, Lucille; Radford, Alan D; Blaxter, Mark L; Tanya, Vincent N; Trees, Alexander J; Cordaux, Richard; Wastling, Jonathan M; Makepeace, Benjamin L

    2012-12-01

    The α-proteobacterium Wolbachia is probably the most prevalent, vertically transmitted symbiont on Earth. In contrast with its wide distribution in arthropods, Wolbachia is restricted to one family of animal-parasitic nematodes, the Onchocercidae. This includes filarial pathogens such as Onchocerca volvulus, the cause of human onchocerciasis, or river blindness. The symbiosis between filariae and Wolbachia is obligate, although the basis of this dependency is not fully understood. Previous studies suggested that Wolbachia may provision metabolites (e.g., haem, riboflavin, and nucleotides) and/or contribute to immune defense. Importantly, Wolbachia is restricted to somatic tissues in adult male worms, whereas females also harbor bacteria in the germline. We sought to characterize the nature of the symbiosis between Wolbachia and O. ochengi, a bovine parasite representing the closest relative of O. volvulus. First, we sequenced the complete genome of Wolbachia strain wOo, which revealed an inability to synthesize riboflavin de novo. Using RNA-seq, we also generated endobacterial transcriptomes from male soma and female germline. In the soma, transcripts for membrane transport and respiration were up-regulated, while the gonad exhibited enrichment for DNA replication and translation. The most abundant Wolbachia proteins, as determined by geLC-MS, included ligands for mammalian Toll-like receptors. Enzymes involved in nucleotide synthesis were dominant among metabolism-related proteins, whereas the haem biosynthetic pathway was poorly represented. We conclude that Wolbachia may have a mitochondrion-like function in the soma, generating ATP for its host. Moreover, the abundance of immunogenic proteins in wOo suggests a role in diverting the immune system toward an ineffective antibacterial response. PMID:22919073

  20. Analysis of gene expression from the Wolbachia genome of a filarial nematode supports both metabolic and defensive roles within the symbiosis

    PubMed Central

    Darby, Alistair C.; Armstrong, Stuart D.; Bah, Germanus S.; Kaur, Gaganjot; Hughes, Margaret A.; Kay, Suzanne M.; Koldkjær, Pia; Rainbow, Lucille; Radford, Alan D.; Blaxter, Mark L.; Tanya, Vincent N.; Trees, Alexander J.; Cordaux, Richard; Wastling, Jonathan M.; Makepeace, Benjamin L.

    2012-01-01

    The α-proteobacterium Wolbachia is probably the most prevalent, vertically transmitted symbiont on Earth. In contrast with its wide distribution in arthropods, Wolbachia is restricted to one family of animal-parasitic nematodes, the Onchocercidae. This includes filarial pathogens such as Onchocerca volvulus, the cause of human onchocerciasis, or river blindness. The symbiosis between filariae and Wolbachia is obligate, although the basis of this dependency is not fully understood. Previous studies suggested that Wolbachia may provision metabolites (e.g., haem, riboflavin, and nucleotides) and/or contribute to immune defense. Importantly, Wolbachia is restricted to somatic tissues in adult male worms, whereas females also harbor bacteria in the germline. We sought to characterize the nature of the symbiosis between Wolbachia and O. ochengi, a bovine parasite representing the closest relative of O. volvulus. First, we sequenced the complete genome of Wolbachia strain wOo, which revealed an inability to synthesize riboflavin de novo. Using RNA-seq, we also generated endobacterial transcriptomes from male soma and female germline. In the soma, transcripts for membrane transport and respiration were up-regulated, while the gonad exhibited enrichment for DNA replication and translation. The most abundant Wolbachia proteins, as determined by geLC-MS, included ligands for mammalian Toll-like receptors. Enzymes involved in nucleotide synthesis were dominant among metabolism-related proteins, whereas the haem biosynthetic pathway was poorly represented. We conclude that Wolbachia may have a mitochondrion-like function in the soma, generating ATP for its host. Moreover, the abundance of immunogenic proteins in wOo suggests a role in diverting the immune system toward an ineffective antibacterial response. PMID:22919073

  1. Distribution patterns of Wolbachia endosymbionts in the closely related flower bugs of the genus Orius: implications for coevolution and horizontal transfer.

    PubMed

    Watanabe, Masaya; Tagami, Yohsuke; Miura, Kazuki; Kageyama, Daisuke; Stouthamer, Richard

    2012-08-01

    Vertical transmission is the primary route of the endosymbiont Wolbachia for its own spread among invertebrate hosts, but horizontal transmission between different hosts is believed to have occurred multiple times. However, it is not well known how Wolbachia commonly spread among closely related hosts. We focused on the closely related species of the minute pirate bugs belonging to the genus Orius, which are important biological control agents in agricultural crops because they are the most useful natural enemy of various tiny pests, such as thrips. Here, we examined five Orius species (Orius sauteri, Orius nagaii, Orius minutus, Orius strigicollis, and Orius tantillus) from eight geographic localities in Japan for Wolbachia infection. Two distinct strains, wOus1 and wOus2, were detected based on Wolbachia surface protein (wsp) gene sequencing. Furthermore, multilocus sequence typing revealed that each of the strains comprised two variants that differed in a single nucleotide. The overall distribution patterns of the two Wolbachia strains were found to differ among host species: prevalent double infection with wOus1 and wOus2 in O. strigicollis; fixation of single infection with wOus2 in O. nagaii; occurrence of single infection with wOus1 in O. sauteri; prevalence of single infection with wOus1 in O. minutus with an exception in a single population; and lack of Wolbachia infection in O. tantillus. Such differences in the distribution patterns of Wolbachia may reflect the evolutionary history of Wolbachia infection among Orius species and/or ecological and physiological differences among the Orius species that determine the invasiveness and maintenance of the two Wolbachia strains. PMID:22450512

  2. Application of wMelPop Wolbachia Strain to Crash Local Populations of Aedes aegypti

    PubMed Central

    Ritchie, Scott A.; Townsend, Michael; Paton, Chris J.; Callahan, Ashley G.; Hoffmann, Ary A.

    2015-01-01

    The endosymbiotic bacteria Wolbachia pipientis (wMel strain) has been successfully established in several populations of Aedes aegypti, the primary dengue vector. The virulent Wolbachia strain wMelPop is known to cause several pathological impacts (increased egg mortality, life shortening, etc.) reducing overall fitness in the mosquito Ae. aegypti. Increased egg mortality could substantially reduce egg banks in areas with a lengthy monsoonal dry season, and be employed to eliminate local populations. We tested this application under semi-field cage conditions. First, we determined that wMelPop infection significantly reduced the survival of desiccation-resistant eggs of the dengue vector Ae. aegypti, with shade and temperature having a significant impact; nearly all wMelPop-infected eggs failed to hatch after 6 and 10 weeks in summer and winter conditions, respectively. In laboratory selection experiments we found that egg desiccation resistance can be increased by selection, and that this effect of wMelPop infection is due to the nuclear background of the host rather than Wolbachia. We then conducted an invasion of wMelPop within a semi-field cage using sustained weekly releases of wMelPop infected mosquitoes, with fixation achieved after 9 weeks. The egg populations wMelPop infected and an uninfected control were then subjected to a simulated prolonged monsoonal dry season (2.5 months) before flooding to induce hatching. The wMelPop infected eggs suffered significantly greater mortality than the controls, with only 0.67% and 4.35% of respective infected and uninfected eggs held in 99% shade hatching after 80 days. These studies suggest that wMelPop could be used to locally eliminate populations of Ae. aegypti that are exposed to prolonged dry conditions, particularly if combined with vector control. PMID:26204449

  3. [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. PMID:23405594

  4. Immune activation by life-shortening Wolbachia and reduced filarial competence in mosquitoes

    PubMed Central

    Kambris, Zakaria; Cook, Peter E.; Phuc, Hoang K.; Sinkins, Steven P.

    2010-01-01

    Wolbachia strain wMelPop reduces longevity of its Drosophila melanogaster host and halves lifespan when introduced into the mosquito Aedes aegypti. We show that wMelPop induces upregulation of the mosquito innate immune system and that its presence inhibits the development of filarial nematodes in the mosquito. These data suggest that wMelPop could be used in the global effort to eliminate lymphatic filariasis, and possibly the control of other mosquito-borne parasites where immune preactivation inhibits their development. The cost of constitutive immune upregulation may contribute to the life-shortening phenotype. PMID:19797660

  5. The virulent Wolbachia strain wMelPop increases the frequency of apoptosis in the female germline cells of Drosophila melanogaster

    PubMed Central

    2012-01-01

    Background Wolbachia are bacterial endosymbionts of many arthropod species in which they manipulate reproductive functions. The distribution of these bacteria in the Drosophila ovarian cells at different stages of oogenesis has been amply described. The pathways along which Wolbachia influences Drosophila oogenesis have been, so far, little studied. It is known that Wolbachia are abundant in the somatic stem cell niche of the Drosophila germarium. A checkpoint, where programmed cell death, or apoptosis, can occur, is located in region 2a/2b of the germarium, which comprises niche cells. Here we address the question whether or not the presence of Wolbachia in germarium cells can affect the frequency of cyst apoptosis in the checkpoint. Results Our current fluorescent microscopic observations showed that the wMel and wMelPop strains had different effects on female germline cells of D. melanogaster. The Wolbachia strain wMel did not affect the frequency of apoptosis in cells of the germarium. The presence of the Wolbachia strain wMelPop in the D. melanogasterw1118 ovaries increased the number of germaria where cells underwent apoptosis in the checkpoint. Based on the appearance in the electron microscope, there was no difference in morphological features of apoptotic cystocytes between Wolbachia-infected and uninfected flies. Bacteria with normal ultrastructure and large numbers of degenerating bacteria were found in the dying cyst cells. Conclusions Our current study demonstrated that the Wolbachia strain wMelPop affects the egg chamber formation in the D. melanogaster ovaries. This led to an increase in the number of germaria containing apoptotic cells. It is suggested that Wolbachia can adversely interfere either with the cystocyte differentiation into the oocyte or with the division of somatic stem cells giving rise to follicle cells and, as a consequence, to improper ratio of germline cells to follicle cells and, ultimately, to apoptosis of cysts. There was no

  6. 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.

  7. Study of LH2 fueled subsonic passenger transport aircraft

    NASA Technical Reports Server (NTRS)

    Brewer, G. D.; Morris, R. E.

    1976-01-01

    The potential of using liquid hydrogen as fuel in subsonic transport aircraft was investigated to explore an expanded matrix of passenger aircraft sizes. Aircraft capable of carrying 130 passengers 2,780 km (1500 n.mi.); 200 passengers 5,560 km (3000 n.mi.); and 400 passengers on a 9,265 km (5000 n.mi.) radius mission, were designed parametrically. Both liquid hydrogen and conventionally fueled versions were generated for each payload/range in order that comparisons could be made. Aircraft in each mission category were compared on the basis of weight, size, cost, energy utilization, and noise.

  8. Wolbachia multilocus sequence typing of singly infected and multiply infected populations of Northern Corn Rootworm (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; multiply-infected. The infected states are associated with different mtDNA haplotypes and reduced mtDNA ...

  9. Wolbachia wsp gene hypervariable region specific PCR primers detect multiple strain infections in northern corn rootworm (Diabrotica barberi)

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The northern corn rootworm (Diabrotica barberi)(Coleoptera: Chrysomelidae) 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 haplo...

  10. Operational use of household bleach to "crash and release" Aedes aegypti prior to Wolbachia-infected mosquito release.

    PubMed

    Jacups, Susan P; Ball, Tamara S; Paton, Christopher J; Johnson, Petrina H; Ritchie, Scott A

    2013-03-01

    Dengue (family Flaviviridae, genus Flavivirus, DENV) remains the leading arboviral cause of mortality in the tropics. Wolbachia pipientis has been shown to interrupt DENV transmission and is presently being trialled as a biological control. However, deployment issues have arisen on methods to temporarily suppress wild mosquito populations before Wolbachia-infected mosquito releases. By suppressing wild populations, fewer Ae. aegypti releases are required to achieve a sustainable Wolbachia density threshold. Furthermore, public distress is reduced. This study tests the application of domestic bleach (4% NaCIO) to temporarily "crash" immature Aedes populations in water-filled containers. Spray application NaClO (215 ppm) resulted in a mean 48-h mortality of 100, 100, 97, and 88% of eggs, second-instar larvae, fourth-instar larvae, and pupae, respectively. In the field, NaClO delayed ovipositing by 9 d in cooler months, and 11 d in hotter months, after which oviposition resumed in treated receptacles. We found bleach treatment of pot-plant bases did not cause wilting, yellowing, or dropping of leaves in two ornamental plants species. Domestically available NaClO could be adopted for a "crash and release" strategy to temporarily suppress wild populations of Ae. aegypti in containers before release of Wolbachia-infected mosquitoes. The "crash and release" strategy is also applicable to other mosquito species, e.g., Aedes albopictus (Skuse), in strategies using released mosquitoes. PMID:23540123

  11. Identifying a set of influential spreaders in complex networks

    PubMed Central

    Zhang, Jian-Xiong; Chen, Duan-Bing; Dong, Qiang; Zhao, Zhi-Dan

    2016-01-01

    Identifying a set of influential spreaders in complex networks plays a crucial role in effective information spreading. A simple strategy is to choose top-r ranked nodes as spreaders according to influence ranking method such as PageRank, ClusterRank and k-shell decomposition. Besides, some heuristic methods such as hill-climbing, SPIN, degree discount and independent set based are also proposed. However, these approaches suffer from a possibility that some spreaders are so close together that they overlap sphere of influence or time consuming. In this report, we present a simply yet effectively iterative method named VoteRank to identify a set of decentralized spreaders with the best spreading ability. In this approach, all nodes vote in a spreader in each turn, and the voting ability of neighbors of elected spreader will be decreased in subsequent turn. Experimental results on four real networks show that under Susceptible-Infected-Recovered (SIR) and Susceptible-Infected (SI) models, VoteRank outperforms the traditional benchmark methods on both spreading rate and final affected scale. What’s more, VoteRank has superior computational efficiency. PMID:27296252

  12. Influential Factors of Z-pin Bridging Force

    NASA Astrophysics Data System (ADS)

    Li, Chenghu; Wu, Zhe; Meng, Zhijun; Li, Muchen

    2014-08-01

    The nine-pin bridging force experiments are conducted at room temperature (20 °C) and at 75 °C. A three-dimensional z-pin unit cell for finite element analysis is established to study the influential factors of the z-pin bridging force. The experimental results show that the z-pin bridging force at 75 °C is smaller than at 20 °C. The z-pin bridging force is highly dependent on the surface area of z-pin. Therefore, it is feasible that the bridging force can be enhanced by changing the surface area of z-pin. The study also shows that the lay-up sequences of laminates have an impact on the z-pin bridging force, and the maximum bridging force of z-pin rises with the increase elastic modules of the resin. The z-pin bridging force reduces with rise of temperature for two reasons: The elastic modulus and shear strength of the resin decrease with rise of temperature, forcing the bridging force to reduce and the resin makes some clamping effect on z-pin, pushing the bridging force to increase.

  13. Identifying a set of influential spreaders in complex networks

    NASA Astrophysics Data System (ADS)

    Zhang, Jian-Xiong; Chen, Duan-Bing; Dong, Qiang; Zhao, Zhi-Dan

    2016-06-01

    Identifying a set of influential spreaders in complex networks plays a crucial role in effective information spreading. A simple strategy is to choose top-r ranked nodes as spreaders according to influence ranking method such as PageRank, ClusterRank and k-shell decomposition. Besides, some heuristic methods such as hill-climbing, SPIN, degree discount and independent set based are also proposed. However, these approaches suffer from a possibility that some spreaders are so close together that they overlap sphere of influence or time consuming. In this report, we present a simply yet effectively iterative method named VoteRank to identify a set of decentralized spreaders with the best spreading ability. In this approach, all nodes vote in a spreader in each turn, and the voting ability of neighbors of elected spreader will be decreased in subsequent turn. Experimental results on four real networks show that under Susceptible-Infected-Recovered (SIR) and Susceptible-Infected (SI) models, VoteRank outperforms the traditional benchmark methods on both spreading rate and final affected scale. What’s more, VoteRank has superior computational efficiency.

  14. Influential parameters on particle exposure of pedestrians in urban microenvironments

    NASA Astrophysics Data System (ADS)

    Buonanno, G.; Fuoco, F. C.; Stabile, L.

    2011-03-01

    Exposure to particle concentrations in urban areas was evaluated in several studies since airborne particles are considered to bring about adverse health effects. Transportation modes and urban microenvironments account for the highest contributions to the overall daily particle exposure concentration. In the present study an evaluation of the influential parameters affecting particle exposure of pedestrian in urban areas is reported. Street geometry, traffic mode, wind speed and direction effects were analyzed through an experimental campaign performed in different streets of an Italian town. To this purpose a high-resolution time measurement apparatus was used in order to capture the dynamic of the freshly emitted particles. Number, surface area and mass concentrations and distributions were measured continuously along both the sides of street canyons and avenue canyons during 10-minutes walking routes. The combined effect of street geometry and wind direction may contribute strongly to dilute the fresh particles emitted by vehicles. In particular, street canyons are characterized by lower ventilation phenomena which lead to similar concentration values on both the side of the street. Higher wind speed was found to decrease concentrations in the canyon. Traffic mode also seems to influence exposure concentrations. In particular, submicrometer particle mass concentration was higher as the traffic is more congested; otherwise, coarse fraction dominates mass exposure concentration along street characterized by a more fluent traffic, showing a typical resuspension modality.

  15. Combination methods for identifying influential nodes in networks

    NASA Astrophysics Data System (ADS)

    Gao, Chao; Zhong, Lu; Li, Xianghua; Zhang, Zili; Shi, Ning

    2015-11-01

    Identifying influential nodes is of theoretical significance in many domains. Although lots of methods have been proposed to solve this problem, their evaluations are under single-source attack in scale-free networks. Meanwhile, some researches have speculated that the combinations of some methods may achieve more optimal results. In order to evaluate this speculation and design a universal strategy suitable for different types of networks under the consideration of multi-source attacks, this paper proposes an attribute fusion method with two independent strategies to reveal the correlation of existing ranking methods and indicators. One is based on feature union (FU) and the other is based on feature ranking (FR). Two different propagation models in the fields of recommendation system and network immunization are used to simulate the efficiency of our proposed method. Experimental results show that our method can enlarge information spreading and restrain virus propagation in the application of recommendation system and network immunization in different types of networks under the condition of multi-source attacks.

  16. Identifying influential user communities on the social network

    NASA Astrophysics Data System (ADS)

    Hu, Weishu; Gong, Zhiguo; Hou U, Leong; Guo, Jingzhi

    2015-10-01

    Nowadays social network services have been popularly used in electronic commerce systems. Users on the social network can develop different relationships based on their common interests and activities. In order to promote the business, it is interesting to explore hidden relationships among users developed on the social network. Such knowledge can be used to locate target users for different advertisements and to provide effective product recommendations. In this paper, we define and study a novel community detection problem that is to discover the hidden community structure in large social networks based on their common interests. We observe that the users typically pay more attention to those users who share similar interests, which enable a way to partition the users into different communities according to their common interests. We propose two algorithms to detect influential communities using common interests in large social networks efficiently and effectively. We conduct our experimental evaluation using a data set from Epinions, which demonstrates that our method achieves 4-11.8% accuracy improvement over the state-of-the-art method.

  17. Identifying a set of influential spreaders in complex networks.

    PubMed

    Zhang, Jian-Xiong; Chen, Duan-Bing; Dong, Qiang; Zhao, Zhi-Dan

    2016-01-01

    Identifying a set of influential spreaders in complex networks plays a crucial role in effective information spreading. A simple strategy is to choose top-r ranked nodes as spreaders according to influence ranking method such as PageRank, ClusterRank and k-shell decomposition. Besides, some heuristic methods such as hill-climbing, SPIN, degree discount and independent set based are also proposed. However, these approaches suffer from a possibility that some spreaders are so close together that they overlap sphere of influence or time consuming. In this report, we present a simply yet effectively iterative method named VoteRank to identify a set of decentralized spreaders with the best spreading ability. In this approach, all nodes vote in a spreader in each turn, and the voting ability of neighbors of elected spreader will be decreased in subsequent turn. Experimental results on four real networks show that under Susceptible-Infected-Recovered (SIR) and Susceptible-Infected (SI) models, VoteRank outperforms the traditional benchmark methods on both spreading rate and final affected scale. What's more, VoteRank has superior computational efficiency. PMID:27296252

  18. Passenger car hydrocarbon emissions speciation. Final report

    SciTech Connect

    Black, F.; High, L.

    1980-05-01

    Emission factors for over 60 individual hydrocarbon compounds were determined for four passenger cars. The cars included a 1963 Chevrolet, a 1977 Mustang, and 1978 Monarch, and 1979 LTD II. The speciation data is reported for both tailpipe and evaporative emissions. The tailpipe emissions were for the urban driving conditions of the Federal Test Procedure used in motor vehicle certification. The evaporative emissions were for both diurnal and hot soak conditions, also prescribed in the Federal Test Procedure for certification. The vehicle tests involved four gasoline fuels of varying composition.

  19. Advanced Vehicle system concepts. [nonpetroleum passenger transportation

    NASA Technical Reports Server (NTRS)

    Hardy, K. S.; Langendoen, J. M.

    1983-01-01

    Various nonpetroleum vehicle system concepts for passenger vehicles in the 1990's are being considered as part of the Advanced Vehicle (AV) Assessment at the Jet Propulsion Laboratory. The vehicle system and subsystem performance requirements, the projected characteristics of mature subsystem candidates, and promising systems are presented. The system candidates include electric and hybrid vehicles powered by electricity with or without a nonpetroleum power source. The subsystem candidates include batteries (aqueous-mobile, flow, high-temperature, and metal-air), fuel cells (phosphoric acid, advanced acids, and solid polymer electrolyte), nonpetroleum heat engines, advanced dc and ac propulsion components, power-peaking devices, and transmissions.

  20. From Lab to Field: The Influence of Urban Landscapes on the Invasive Potential of Wolbachia in Brazilian Aedes aegypti Mosquitoes

    PubMed Central

    Caragata, Eric Pearce; Silva, Jéssica Barreto Lopes; Villela, Daniel Antunes Maciel; Maciel-de-Freitas, Rafael; Moreira, Luciano Andrade

    2015-01-01

    Background The symbiotic bacterium Wolbachia is currently being trialled as a biocontrol agent in several countries to reduce dengue transmission. Wolbachia can invade and spread to infect all individuals within wild mosquito populations, but requires a high rate of maternal transmission, strong cytoplasmic incompatibility and low fitness costs in the host in order to do so. Additionally, extensive differences in climate, field-release protocols, urbanization level and human density amongst the sites where this bacterium has been deployed have limited comparison and analysis of Wolbachia’s invasive potential. Methodology/Principal Findings We examined key phenotypic effects of the wMel Wolbachia strain in laboratory Aedes aegypti mosquitoes with a Brazilian genetic background to characterize its invasive potential. We show that the wMel strain causes strong cytoplasmic incompatibility, a high rate of maternal transmission and has no evident detrimental effect on host fecundity or fertility. Next, to understand the effects of different urban landscapes on the likelihood of mosquito survival, we performed mark-release-recapture experiments using Wolbachia-uninfected Brazilian mosquitoes in two areas of Rio de Janeiro where Wolbachia will be deployed in the future. We characterized the mosquito populations in relation to the socio-demographic conditions at these sites, and at three other future release areas. We then constructed mathematical models using both the laboratory and field data, and used these to describe the influence of urban environmental conditions on the likelihood that the Wolbachia infection frequency could reach 100% following mosquito release. We predict successful invasion at all five field sites, however the conditions by which this occurs vary greatly between sites, and are strongly influenced by the size of the local mosquito population. Conclusions/Significance Through analysis of laboratory, field and mathematical data, we show that the w

  1. 49 CFR 238.307 - Periodic mechanical inspection of passenger cars and unpowered vehicles used in passenger trains.

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    ... TRANSPORTATION PASSENGER EQUIPMENT SAFETY STANDARDS Inspection, Testing, and Maintenance Requirements for Tier I... (iii) Each railroad shall retain records of the inspection, testing, and maintenance of the...

  2. Characterization of intersex production in Trichogramma kaykai infected with parthenogenesis-inducing Wolbachia

    NASA Astrophysics Data System (ADS)

    Tulgetske, Genet M.; Stouthamer, Richard

    2012-02-01

    Sexually aberrant individuals, displaying both male and female characteristics, are rare in occurrence but are documented throughout the animal kingdom. In parasitoid wasps of the genus Trichogramma, such individuals typically appear as a result of rearing Wolbachia-infected thelytokous wasps at high temperatures. Sexually aberrant Trichogramma have been referred to interchangeably in the literature as gynandromorphs, sexual mosaics and intersexes. However, accurately used, the terms "gynandromorph" and "sexual mosaic" describe an individual composed of a mixture of genetically distinct tissues corresponding to the sexual phenotypes observed, while "intersex" refers to an individual having a uniform genetic constitution but with some tissues exhibiting sexual phenotypes conflicting with the associated genotype. Here, we investigate the heat-induced production of sexually aberrant offspring by thelytokous Trichogramma kaykai. Aberrant individuals were rare, but each was characterized as one of 11 morphotypes ranging from very feminine to very masculine. Overall, the production of aberrant individuals increased with time from the onset of maternal oviposition. However, while the production of males also increased with time, the degree of masculinity of aberrant individuals did not; the different morphotypes appeared to be produced haphazardly. We conclude that the aberrant individuals produced by T. kaykai are actually intersexes and not gynandromorphs. The wasp's close association with Wolbachia and the absence of intersexes in uninfected populations allow us to discuss a possible origin of the condition.

  3. Anti-Wolbachia drug discovery and development: safe macrofilaricides for onchocerciasis and lymphatic filariasis.

    PubMed

    Taylor, Mark J; Hoerauf, Achim; Townson, Simon; Slatko, Barton E; Ward, Stephen A

    2014-01-01

    Anti-Wolbachia therapy delivers safe macrofilaricidal activity with superior therapeutic outcomes compared to all standard anti-filarial treatments, with the added benefit of substantial improvements in clinical pathology. These outcomes can be achieved, in principle, with existing registered drugs, e.g. doxycycline, that are affordable, available to endemic communities and have well known, albeit population-limiting, safety profiles. The key barriers to using doxycycline as an mass drug administration (MDA) strategy for widespread community-based control are the logistics of a relatively lengthy course of treatment (4-6 weeks) and contraindications in children under eight years and pregnancy. Therefore, the primary goal of the anti-Wolbachia (A·WOL) consortium is to find drugs and regimens that reduce the period of treatment from weeks to days (7 days or less), and to find drugs which would be safe in excluded target populations (pregnancy and children). A secondary goal is to refine regimens of existing antibiotics suitable for a more restricted use, prior to the availability of a regimen that is compatible with MDA usage. For example, for use in the event of the emergence of drug-resistance, in individuals with high loiasis co-infection and at risk of severe adverse events (SAE) to ivermectin, or in post-MDA 'endgame scenarios', where test and treat strategies become more cost effective and deliverable. PMID:23866958

  4. Coexistence of Wolbachia with Buchnera aphidicola and a Secondary Symbiont in the Aphid Cinara cedri

    PubMed Central

    Gómez-Valero, Laura; Soriano-Navarro, Mario; Pérez-Brocal, Vicente; Heddi, Abdelaziz; Moya, Andrés; García-Verdugo, José Manuel; Latorre, Amparo

    2004-01-01

    Intracellular symbiosis is very common in the insect world. For the aphid Cinara cedri, we have identified by electron microscopy three symbiotic bacteria that can be characterized by their different sizes, morphologies, and electrodensities. PCR amplification and sequencing of the 16S ribosomal DNA (rDNA) genes showed that, in addition to harboring Buchnera aphidicola, the primary endosymbiont of aphids, C. cedri harbors a secondary symbiont (S symbiont) that was previously found to be associated with aphids (PASS, or R type) and an α-proteobacterium that belongs to the Wolbachia genus. Using in situ hybridization with specific bacterial probes designed for symbiont 16S rDNA sequences, we have shown that Wolbachia was represented by only a few minute bacteria surrounding the S symbionts. Moreover, the observed B. aphidicola and the S symbionts had similar sizes and were housed in separate specific bacterial cells, the bacteriocytes. Interestingly, in contrast to the case for all aphids examined thus far, the S symbionts were shown to occupy a similarly sized or even larger bacteriocyte space than B. aphidicola. These findings, along with the facts that C. cedri harbors the B. aphidicola strain with the smallest bacterial genome and that the S symbionts infect all Cinara spp. analyzed so far, suggest the possibility of bacterial replacement in these species. PMID:15375144

  5. Variations of immune parameters in terrestrial isopods: a matter of gender, aging and Wolbachia.

    PubMed

    Sicard, Mathieu; Chevalier, Frédéric; De Vlechouver, Mickaël; Bouchon, Didier; Grève, Pierre; Braquart-Varnier, Christine

    2010-09-01

    Ecological factors modulate animal immunocompetence and potentially shape the evolution of their immune systems. Not only environmental parameters impact on immunocompetence: Aging is one major cause of variability of immunocompetence between individuals, and sex-specific levels of immunocompetence have also been frequently described. Moreover, a growing core of data put in light that vertically transmitted symbionts can dramatically modulate the immunocompetence of their hosts. In this study, we addressed the influence of gender, age and the feminising endosymbiont Wolbachia (wVulC) on variations in haemocyte density, total PO activity and bacterial load in the haemolymph of the terrestrial isopod Armadillidium vulgare. This host-symbiont system is of particular interest to address this question since: (1) wVulC was previously shown as immunosuppressive in middle-aged females and (2) wVulC influences sex determination. We show that age, gender and Wolbachia modulate together immune parameters in A. vulgare. However, wVulC, which interacts with aging, appears to be the prominent factor interfering with both PO activity and haemocyte density. This interference with immune parameters is not the only aspect of wVulC virulence on its host, as reproduction and survival are also altered. PMID:20676599

  6. Variations of immune parameters in terrestrial isopods: a matter of gender, aging and Wolbachia

    NASA Astrophysics Data System (ADS)

    Sicard, Mathieu; Chevalier, Frédéric; de Vlechouver, Mickaël; Bouchon, Didier; Grève, Pierre; Braquart-Varnier, Christine

    2010-09-01

    Ecological factors modulate animal immunocompetence and potentially shape the evolution of their immune systems. Not only environmental parameters impact on immunocompetence: Aging is one major cause of variability of immunocompetence between individuals, and sex-specific levels of immunocompetence have also been frequently described. Moreover, a growing core of data put in light that vertically transmitted symbionts can dramatically modulate the immunocompetence of their hosts. In this study, we addressed the influence of gender, age and the feminising endosymbiont Wolbachia ( wVulC) on variations in haemocyte density, total PO activity and bacterial load in the haemolymph of the terrestrial isopod Armadillidium vulgare. This host-symbiont system is of particular interest to address this question since: (1) wVulC was previously shown as immunosuppressive in middle-aged females and (2) wVulC influences sex determination. We show that age, gender and Wolbachia modulate together immune parameters in A. vulgare. However, wVulC, which interacts with aging, appears to be the prominent factor interfering with both PO activity and haemocyte density. This interference with immune parameters is not the only aspect of wVulC virulence on its host, as reproduction and survival are also altered.

  7. Crystallization and preliminary diffraction analysis of a DsbA homologue from Wolbachia pipientis

    SciTech Connect

    Kurz, M.; Iturbe-Ormaetxe, I.; Jarrott, R.; O’Neill, S. L.; Byriel, K. A.; Martin, J. L. Heras, B.

    2008-02-01

    The first crystallization of a W. pipientis protein, α-DsbA1, was achieved using hanging-drop and sitting-drop vapour diffusion. α-DsbA1 is one of two DsbA homologues encoded by the Gram-negative α-proteobacterium Wolbachia pipientis, an endosymbiont that can behave as a reproductive parasite in insects and as a mutualist in medically important filarial nematodes. The α-DsbA1 protein is thought to be important for the folding and secretion of Wolbachia proteins involved in the induction of reproductive distortions. Crystals of native and SeMet α-DsbA1 were grown by vapour diffusion and belong to the monoclinic space group C2, with unit-cell parameters a = 71.4, b = 49.5, c = 69.3 Å, β = 107.0° and one molecule in the asymmetric unit (44% solvent content). X-ray data were recorded from native crystals to a resolution of 2.01 Å using a copper anode and data from SeMet α-DsbA1 crystals were recorded to 2.45 Å resolution using a chromium anode.

  8. Incidence of the endosymbionts Wolbachia, Cardinium and Spiroplasma in phytoseiid mites and associated prey.

    PubMed

    Enigl, Monika; Schausberger, Peter

    2007-01-01

    Endosymbiotic bacteria that potentially influence reproduction and other fitness-related traits of their hosts are widespread in insects and mites and their appeal to researchers' interest is still increasing. We screened 20 strains of 12 agriculturally relevant herbivorous and predatory mite species for infection with Wolbachia, Cardinium and Spiroplasma by the use of PCR. The majority of specimens originated from Austria and were field collected or mass-reared. Eight out of 20 strains (40%) tested, representing seven of 12 mite species (58%), carried at least one of the three bacteria. We found Wolbachia in the herbivorous spider mites Tetranychus urticae and Bryobia rubrioculus, with the former also carrying Spiroplasma and the latter also carrying Cardinium. Cardinium was furthermore found in two populations of the predatory mite Euseius finlandicus and the spider mite Eotetranychus uncatus. Spiroplasma was detected in the predatory mite Neoseiulus californicus. All bacteria positive PCR products were sequenced, submitted to GenBank and analyzed in BLAST queries. We found high similarities to complete identity with bacteria found in the same and different mite species but also with bacteria found in insect species like ladybirds, butterflies and minute pirate bugs, Orius. We discuss the significance of potential (multiple) infections with the investigated bacteria for biological control. PMID:17554631

  9. The microbiome of Folsomia candida: an assessment of bacterial diversity in a Wolbachia-containing animal.

    PubMed

    Agamennone, Valeria; Jakupović, Dennis; Weedon, James T; Suring, Wouter J; van Straalen, Nico M; Roelofs, Dick; Röling, Wilfred F M

    2015-11-01

    The springtail Folsomia candida is an important model organism for soil ecology, ecotoxicology and ecogenomics. The decomposer activities of soil invertebrates like Folsomia depend on their relationship with microbial communities including gut symbionts. In this paper, we apply high-throughput sequencing to provide a detailed characterization of the bacterial community associated with parthenogenetic F. candida. First, we evaluated a method to suppress the amplification of DNA from the endosymbiont Wolbachia, to prevent it from interfering with the identification of less abundant operational taxonomic units (OTUs). The suppression treatment applied was effective against Wolbachia and did not interfere with the detection of the most abundant OTUs (59 OTUs, contributing over 87% of the reads). However, this method did affect the inferred community composition. Significant differences were subsequently observed in the composition of bacterial communities associated with two different strains of F. candida. A total of 832 OTUs were found, of which 45% were only present in one strain and 17% only in the other. Among the 20 most abundant OTUs, 16 were shared between strains. Denaturing gradient gel electrophoresis and clone libraries, although unable to capture the full diversity of the bacterial community, provided results that supported the NGS data. PMID:26499484

  10. The Wolbachia WO bacteriophage proteome in the Aedes albopictus C/wStr1 cell line: evidence for lytic activity?

    PubMed

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

    2016-01-01

    Wolbachia pipientis (Rickettsiales), an obligate intracellular alphaproteobacterium in insects, manipulates host reproduction to maximize invasion of uninfected insect populations. Modification of host population structure has potential applications for control of pest species, particularly if Wolbachia can be maintained, manipulated, and genetically engineered in vitro. Although Wolbachia maintains an obligate mutualism with genome stability in nematodes, arthropods can be co-infected with distinct Wolbachia strains, and horizontal gene transfer between strains is potentially mediated by WO phages encoded within Wolbachia genomes. Proteomic analysis of a robust, persistent infection of a mosquito cell line with wStr from the planthopper, Laodelphax striatellus, revealed expression of a full array of WO phage genes, as well as nine of ten non-phage genes that occur between two distinct clusters of WOMelB genes in the genome of wMel, which infects Drosophila melanogaster. These non-phage genes encode potential host-adaptive proteins and are expressed in wStr at higher levels than phage structural proteins. A subset of seven of the non-phage genes is flanked by highly conserved non-coding sequences, including a putative promoter element, that are not present in a syntenically arranged array of homologs in plasmids from three tick-associated Rickettsia spp. These studies expand our understanding of wStr in a host cell line derived from the mosquito, Aedes albopictus, and provide a basis for investigating conditions that favor the lytic phase of the WO phage life cycle and recovery of infectious phage particles. PMID:26427709

  11. Establishment of a Wolbachia Superinfection in Aedes aegypti Mosquitoes as a Potential Approach for Future Resistance Management

    PubMed Central

    De Bruyne, Jyotika Taneja; Kien, Duong Hue T.; Hoang, Nhat Le Thanh; Chau, Nguyen Van Vinh; Iturbe-Ormaetxe, Iñaki; Simmons, Cameron P.; O’Neill, Scott L.

    2016-01-01

    Wolbachia pipientis is an endosymbiotic bacterium estimated to chronically infect between 40–75% of all arthropod species. Aedes aegypti, the principle mosquito vector of dengue virus (DENV), is not a natural host of Wolbachia. The transinfection of Wolbachia strains such as wAlbB, wMel and wMelPop-CLA into Ae. aegypti has been shown to significantly reduce the vector competence of this mosquito for a range of human pathogens in the laboratory. This has led to wMel-transinfected Ae. aegypti currently being released in five countries to evaluate its effectiveness to control dengue disease in human populations. Here we describe the generation of a superinfected Ae. aegypti mosquito line simultaneously infected with two avirulent Wolbachia strains, wMel and wAlbB. The line carries a high overall Wolbachia density and tissue localisation of the individual strains is very similar to each respective single infected parental line. The superinfected line induces unidirectional cytoplasmic incompatibility (CI) when crossed to each single infected parental line, suggesting that the superinfection would have the capacity to replace either of the single constituent infections already present in a mosquito population. No significant differences in fitness parameters were observed between the superinfected line and the parental lines under the experimental conditions tested. Finally, the superinfected line blocks DENV replication more efficiently than the single wMel strain when challenged with blood meals from viremic dengue patients. These results suggest that the deployment of superinfections could be used to replace single infections and may represent an effective strategy to help manage potential resistance by DENV to field deployments of single infected strains. PMID:26891349

  12. The hitchhiker's guide to Europe: the infection dynamics of an ongoing Wolbachia invasion and mitochondrial selective sweep in Rhagoletis cerasi.

    PubMed

    Schuler, Hannes; Köppler, Kirsten; Daxböck-Horvath, Sabine; Rasool, Bilal; Krumböck, Susanne; Schwarz, Dietmar; Hoffmeister, Thomas S; Schlick-Steiner, Birgit C; Steiner, Florian M; Telschow, Arndt; Stauffer, Christian; Arthofer, Wolfgang; Riegler, Markus

    2016-04-01

    Wolbachia is a maternally inherited and ubiquitous endosymbiont of insects. It can hijack host reproduction by manipulations such as cytoplasmic incompatibility (CI) to enhance vertical transmission. Horizontal transmission of Wolbachia can also result in the colonization of new mitochondrial lineages. In this study, we present a 15-year-long survey of Wolbachia in the cherry fruit fly Rhagoletis cerasi across Europe and the spatiotemporal distribution of two prevalent strains, wCer1 and wCer2, and associated mitochondrial haplotypes in Germany. Across most of Europe, populations consisted of either 100% singly (wCer1) infected individuals with haplotype HT1, or 100% doubly (wCer1&2) infected individuals with haplotype HT2, differentiated only by a single nucleotide polymorphism. In central Germany, singly infected populations were surrounded by transitional populations, consisting of both singly and doubly infected individuals, sandwiched between populations fixed for wCer1&2. Populations with fixed infection status showed perfect association of infection and mitochondria, suggesting a recent CI-driven selective sweep of wCer2 linked with HT2. Spatial analysis revealed a range expansion for wCer2 and a large transition zone in which wCer2 splashes appeared to coalesce into doubly infected populations. Unexpectedly, the transition zone contained a large proportion (22%) of wCer1&2 individuals with HT1, suggesting frequent intraspecific horizontal transmission. However, this horizontal transmission did not break the strict association between infection types and haplotypes in populations outside the transition zone, suggesting that this horizontally acquired Wolbachia infection may be transient. Our study provides new insights into the rarely studied Wolbachia invasion dynamics in field populations. PMID:26846713

  13. Identification and Characterization of a Candidate Wolbachia pipientis Type IV Effector That Interacts with the Actin Cytoskeleton

    PubMed Central

    Sheehan, Kathy B.; Martin, MaryAnn; Lesser, Cammie F.; Isberg, Ralph R.

    2016-01-01

    ABSTRACT Many bacteria live as intracellular symbionts, causing persistent infections within insects. One extraordinarily common infection is that of Wolbachia pipientis, which infects 40% of insect species and induces reproductive effects. The bacteria are passed from generation to generation both vertically (through the oocyte) and horizontally (by environmental transmission). Maintenance of the infection within Drosophila melanogaster is sensitive to the regulation of actin, as Wolbachia inefficiently colonizes strains hemizygous for the profilin or villin genes. Therefore, we hypothesized that Wolbachia must depend on the host actin cytoskeleton. In this study, we identify and characterize a Wolbachia protein (WD0830) that is predicted to be secreted by the bacterial parasite. Expression of WD0830 in a model eukaryote (the yeast Saccharomyces cerevisiae) induces a growth defect associated with the appearance of aberrant, filamentous structures which colocalize with rhodamine-phalloidin-stained actin. Purified WD0830 bundles actin in vitro and cosediments with actin filaments, suggesting a direct interaction of the two proteins. We characterized the expression of WD0830 throughout Drosophila development and found it to be upregulated in third-instar larvae, peaking in early pupation, during the critical formation of adult tissues, including the reproductive system. In transgenic flies, heterologously expressed WD0830 localizes to the developing oocyte. Additionally, overexpression of WD0830 results in increased Wolbachia titers in whole flies, in stage 9 and 10 oocytes, and in embryos, compared to controls, suggesting that the protein may facilitate Wolbachia’s replication or transmission. Therefore, this candidate secreted effector may play a role in Wolbachia’s infection of and persistence within host niches. PMID:27381293

  14. Phylogenomics of the Reproductive Parasite Wolbachia pipientis wMel: A Streamlined Genome Overrun by Mobile Genetic Elements

    PubMed Central

    2004-01-01

    The complete sequence of the 1,267,782 bp genome of Wolbachia pipientis wMel, an obligate intracellular bacteria of Drosophila melanogaster, has been determined. Wolbachia, which are found in a variety of invertebrate species, are of great interest due to their diverse interactions with different hosts, which range from many forms of reproductive parasitism to mutualistic symbioses. Analysis of the wMel genome, in particular phylogenomic comparisons with other intracellular bacteria, has revealed many insights into the biology and evolution of wMel and Wolbachia in general. For example, the wMel genome is unique among sequenced obligate intracellular species in both being highly streamlined and containing very high levels of repetitive DNA and mobile DNA elements. This observation, coupled with multiple evolutionary reconstructions, suggests that natural selection is somewhat inefficient in wMel, most likely owing to the occurrence of repeated population bottlenecks. Genome analysis predicts many metabolic differences with the closely related Rickettsia species, including the presence of intact glycolysis and purine synthesis, which may compensate for an inability to obtain ATP directly from its host, as Rickettsia can. Other discoveries include the apparent inability of wMel to synthesize lipopolysaccharide and the presence of the most genes encoding proteins with ankyrin repeat domains of any prokaryotic genome yet sequenced. Despite the ability of wMel to infect the germline of its host, we find no evidence for either recent lateral gene transfer between wMel and D. melanogaster or older transfers between Wolbachia and any host. Evolutionary analysis further supports the hypothesis that mitochondria share a common ancestor with the α-Proteobacteria, but shows little support for the grouping of mitochondria with species in the order Rickettsiales. With the availability of the complete genomes of both species and excellent genetic tools for the host, the w

  15. Establishment of a Wolbachia Superinfection in Aedes aegypti Mosquitoes as a Potential Approach for Future Resistance Management.

    PubMed

    Joubert, D Albert; Walker, Thomas; Carrington, Lauren B; De Bruyne, Jyotika Taneja; Kien, Duong Hue T; Hoang, Nhat Le Thanh; Chau, Nguyen Van Vinh; Iturbe-Ormaetxe, Iñaki; Simmons, Cameron P; O'Neill, Scott L

    2016-02-01

    Wolbachia pipientis is an endosymbiotic bacterium estimated to chronically infect between 40-75% of all arthropod species. Aedes aegypti, the principle mosquito vector of dengue virus (DENV), is not a natural host of Wolbachia. The transinfection of Wolbachia strains such as wAlbB, wMel and wMelPop-CLA into Ae. aegypti has been shown to significantly reduce the vector competence of this mosquito for a range of human pathogens in the laboratory. This has led to wMel-transinfected Ae. aegypti currently being released in five countries to evaluate its effectiveness to control dengue disease in human populations. Here we describe the generation of a superinfected Ae. aegypti mosquito line simultaneously infected with two avirulent Wolbachia strains, wMel and wAlbB. The line carries a high overall Wolbachia density and tissue localisation of the individual strains is very similar to each respective single infected parental line. The superinfected line induces unidirectional cytoplasmic incompatibility (CI) when crossed to each single infected parental line, suggesting that the superinfection would have the capacity to replace either of the single constituent infections already present in a mosquito population. No significant differences in fitness parameters were observed between the superinfected line and the parental lines under the experimental conditions tested. Finally, the superinfected line blocks DENV replication more efficiently than the single wMel strain when challenged with blood meals from viremic dengue patients. These results suggest that the deployment of superinfections could be used to replace single infections and may represent an effective strategy to help manage potential resistance by DENV to field deployments of single infected strains. PMID:26891349

  16. Evidence for female mortality in Wolbachia-mediated cytoplasmic incompatibility in haplodiploid insects: epidemiologic and evolutionary consequences.

    PubMed

    Vavre, F; Fleury, F; Varaldi, J; Fouillet, P; Boulétreau, M

    2000-02-01

    Until now, only two Wolbachia-mediated cytoplasmic incompatibility (CI) types have been described in haplodiploid species, the first in Nasonia (Insect) and the second in Tetranychus (Acari). They both induce a male-biased sex ratio in the incompatible cross. In Nasonia, CI does not reduce fertility since incompatible eggs develop as haploid males, whereas in Tetranychus CI leads to a partial mortality of incompatible eggs, thus reducing the fertility of females. Here, we study Wolbachia infection in a Drosophila parasitoid, Leptopilina heterotoma (Hymenoptera: Figitidae). A survey of Wolbachia infection shows that all natural populations tested are totally infected. Crosses between infected males and cured females show complete incompatibility: almost no females are produced. Moreover, incompatible eggs die early during their development, unlike Nasonia. This early death allows the parasitized Drosophila larva to achieve its development and to emerge. Thus, uninfected females crossed with infected males have reduced offspring production consisting only of males. Evidence of this CI type in insects demonstrates that the difference in CI types of Nasonia and Tetranychus is not due to specific factors of insects or acari. Using theoretical models, we compare the invasion processes of different strategies of Wolbachia: CI in diploid species, the two CI types in haplodiploid species, and parthenogenesis (the classical effect in haplodiploid species). Models show that CI in haplodiploid species is less efficient than in diploid ones. However, the Leptopilina type is advantageous compared to the Nasonia type. Parthenogenesis may be more or less advantageous, depending on the infection cost and on the proportion of fertilized eggs. Finally, we can propose different processes of Wolbachia strategy evolution in haplodiploid species from Nasonia CI type to Leptopilina CI type or parthenogenesis. PMID:10937195

  17. Wolbachia Infections Mimic Cryptic Speciation in Two Parasitic Butterfly Species, Phengaris teleius and P. nausithous (Lepidoptera: Lycaenidae)

    PubMed Central

    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. PMID:24223136

  18. Deceptive single-locus taxonomy and phylogeography: Wolbachia-associated divergence in mitochondrial DNA is not reflected in morphology and nuclear markers in a butterfly species

    PubMed Central

    Kodandaramaiah, Ullasa; Simonsen, Thomas J; Bromilow, Sean; Wahlberg, Niklas; Sperling, Felix

    2013-01-01

    The satyrine butterfly Coenonympha tullia (Nymphalidae: Satyrinae) displays a deep split between two mitochondrial clades, one restricted to northern Alberta, Canada, and the other found throughout Alberta and across North America. We confirm this deep divide and test hypotheses explaining its phylogeographic structure. Neither genitalia morphology nor nuclear gene sequence supports cryptic species as an explanation, instead indicating differences between nuclear and mitochondrial genome histories. Sex-biased dispersal is unlikely to cause such mito-nuclear differences; however, selective sweeps by reproductive parasites could have led to this conflict. About half of the tested samples were infected by Wolbachia bacteria. Using multilocus strain typing for three Wolbachia genes, we show that the divergent mitochondrial clades are associated with two different Wolbachia strains, supporting the hypothesis that the mito-nuclear differences resulted from selection on the mitochondrial genome due to selective sweeps by Wolbachia strains. PMID:24455146

  19. Relative utility of arrhenotokous and Wolbachia-associated thelytokous Odontosema anastrephae figitid fruit fly parasitoids for mass rearing and biological control

    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 reproducti...

  20. Identifying highly influential nodes in the complicated grief network.

    PubMed

    Robinaugh, Donald J; Millner, Alexander J; McNally, Richard J

    2016-08-01

    The network approach to psychopathology conceptualizes mental disorders as networks of mutually reinforcing nodes (i.e., symptoms). Researchers adopting this approach have suggested that network topology can be used to identify influential nodes, with nodes central to the network having the greatest influence on the development and maintenance of the disorder. However, because commonly used centrality indices do not distinguish between positive and negative edges, they may not adequately assess the nature and strength of a node's influence within the network. To address this limitation, we developed 2 indices of a node's expected influence (EI) that account for the presence of negative edges. To evaluate centrality and EI indices, we simulated single-node interventions on randomly generated networks. In networks with exclusively positive edges, centrality and EI were both strongly associated with observed node influence. In networks with negative edges, EI was more strongly associated with observed influence than was centrality. We then used data from a longitudinal study of bereavement to examine the association between (a) a node's centrality and EI in the complicated grief (CG) network and (b) the strength of association between change in that node and change in the remainder of the CG network from 6- to 18-months postloss. Centrality and EI were both correlated with the strength of the association between node change and network change. Together, these findings suggest high-EI nodes, such as emotional pain and feelings of emptiness, may be especially important to the etiology and treatment of CG. (PsycINFO Database Record PMID:27505622

  1. 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…

  2. 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...

  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. 14 CFR 29.807 - Passenger emergency exits.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... 14 Aeronautics and Space 1 2012-01-01 2012-01-01 false Passenger emergency exits. 29.807 Section 29.807 Aeronautics and Space FEDERAL AVIATION ADMINISTRATION, DEPARTMENT OF TRANSPORTATION AIRCRAFT AIRWORTHINESS STANDARDS: TRANSPORT CATEGORY ROTORCRAFT Design and Construction Personnel and Cargo Accommodations § 29.807 Passenger emergency exits....

  5. 14 CFR 121.573 - Briefing passengers: Extended overwater operations.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 14 Aeronautics and Space 3 2010-01-01 2010-01-01 false Briefing passengers: Extended overwater... OPERATING REQUIREMENTS: DOMESTIC, FLAG, AND SUPPLEMENTAL OPERATIONS Flight Operations § 121.573 Briefing passengers: Extended overwater operations. (a) In addition to the oral briefing required by §...

  6. 49 CFR 223.15 - Requirements for existing passenger cars.

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ... regulations of the Office of the Secretary of Transportation, 49 CFR 1.49(m)) ... 49 Transportation 4 2013-10-01 2013-10-01 false Requirements for existing passenger cars. 223.15... ADMINISTRATION, DEPARTMENT OF TRANSPORTATION SAFETY GLAZING STANDARDS-LOCOMOTIVES, PASSENGER CARS AND...

  7. 49 CFR 223.15 - Requirements for existing passenger cars.

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    ... regulations of the Office of the Secretary of Transportation, 49 CFR 1.49(m)) [44 FR 77352, Dec. 31, 1979, as... 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...

  8. 49 CFR 223.15 - Requirements for existing passenger cars.

    Code of Federal Regulations, 2012 CFR

    2012-10-01

    ... regulations of the Office of the Secretary of Transportation, 49 CFR 1.49(m)) ... 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...

  9. 32 CFR 700.842 - Authority over passengers.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... regulations or in orders from competent authority, all passengers in a ship or aircraft of the naval service... and routine of the ship or aircraft. The commanding officer of such ship or aircraft shall take no... aircraft or of any persons embarked, subject a passenger not in the naval service to such restraint as...

  10. 78 FR 71785 - Passenger Train Emergency Systems II

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-11-29

    ... FRA's January 3, 2012, proposed rule on passenger train emergency systems, see 77 FR 153, FRA issues... the Act, FRA published the Passenger Train Emergency Preparedness (PTEP) final rule. See 63 FR 24629...) final rule. See 64 FR 25540. The rule established comprehensive safety standards for railroad...

  11. 49 CFR 223.8 - Additional requirements for passenger equipment.

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ... 49 Transportation 4 2010-10-01 2010-10-01 false Additional requirements for passenger equipment. 223.8 Section 223.8 Transportation Other Regulations Relating to Transportation (Continued) FEDERAL... AND CABOOSES Specific Requirements § 223.8 Additional requirements for passenger equipment....

  12. 46 CFR 178.215 - Weight of passengers and crew.

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ... 46 Shipping 7 2011-10-01 2011-10-01 false Weight of passengers and crew. 178.215 Section 178.215... Weight of passengers and crew. (a) This section applies to each vessel, regardless of when constructed... required, the owner of each vessel must provide the master with the total test weight used in...

  13. 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.

  14. 49 CFR 223.15 - Requirements for existing passenger cars.

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ... regulations of the Office of the Secretary of Transportation, 49 CFR 1.49(m)) ... 49 Transportation 4 2011-10-01 2011-10-01 false Requirements for existing passenger cars. 223.15... ADMINISTRATION, DEPARTMENT OF TRANSPORTATION SAFETY GLAZING STANDARDS-LOCOMOTIVES, PASSENGER CARS AND...

  15. 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.

  16. 14 CFR 221.30 - Passenger fares and charges.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 14 Aeronautics and Space 4 2010-01-01 2010-01-01 false Passenger fares and charges. 221.30 Section 221.30 Aeronautics and Space OFFICE OF THE SECRETARY, DEPARTMENT OF TRANSPORTATION (AVIATION PROCEEDINGS) ECONOMIC REGULATIONS TARIFFS Manner of Filing Tariffs § 221.30 Passenger fares and charges....

  17. 49 CFR 541.5 - Requirements for passenger motor vehicles.

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    ... 49 Transportation 6 2014-10-01 2014-10-01 false Requirements for passenger motor vehicles. 541.5 Section 541.5 Transportation Other Regulations Relating to Transportation (Continued) NATIONAL HIGHWAY TRAFFIC SAFETY ADMINISTRATION, DEPARTMENT OF TRANSPORTATION FEDERAL MOTOR VEHICLE THEFT PREVENTION STANDARD § 541.5 Requirements for passenger...

  18. 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 Section 541.5 Transportation Other Regulations Relating to Transportation (Continued) NATIONAL HIGHWAY TRAFFIC SAFETY ADMINISTRATION, DEPARTMENT OF TRANSPORTATION FEDERAL MOTOR VEHICLE THEFT PREVENTION STANDARD § 541.5 Requirements for passenger...

  19. 49 CFR 541.5 - Requirements for passenger motor vehicles.

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ... confidential treatment pursuant to 49 CFR part 512, the manufacturer must also submit a complete copy of the... 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. 49 CFR 541.5 - Requirements for passenger motor vehicles.

    Code of Federal Regulations, 2012 CFR

    2012-10-01

    ... 49 Transportation 6 2012-10-01 2012-10-01 false Requirements for passenger motor vehicles. 541.5 Section 541.5 Transportation Other Regulations Relating to Transportation (Continued) NATIONAL HIGHWAY TRAFFIC SAFETY ADMINISTRATION, DEPARTMENT OF TRANSPORTATION FEDERAL MOTOR VEHICLE THEFT PREVENTION STANDARD § 541.5 Requirements for passenger...