Palaeosymbiosis Revealed by Genomic Fossils of Wolbachia in a Strongyloidean Nematode

PubMed Central

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

2014-01-01

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

Negative evidence of Wolbachia in the predaceous mite Phytoseiulus persimilis.

PubMed

Enigl, M; Zchori-Fein, E; Schausberger, P

2005-01-01

The cytoplasmically inherited bacterium Wolbachia is widespread in arthropod species and has been repeatedly detected in the predaceous mite Phytoseiulus persimilis. Our original goal was to assess the prevalence of Wolbachia infection in P. persimilis and the potential fitness consequences for this host. To accomplish that goal, seven P. persimilis strains were obtained from Europe, Africa and the USA and reared on the phytophagous mite Tetranychus urticae as prey. After preliminary results showed that the T. urticae used was infected with Wolbachia, the minimum starvation time of the predators to prevent false positive results from undigested prey was determined. We tested DNA samples by PCR (polymerase chain reaction) after starving the predators or feeding them Wolbachia-free T. urticae for various periods. Those experiments showed that Wolbachia could not be detected after 16 h at 25 degrees C and 48 h at 20 degrees C. To verify the results of the PCR analyses, we furthermore conducted crossing experiments with antibiotic-treated and untreated individuals. No indications of Wolbachia effects were recorded. Additionally, we screened live eggs of four of the seven strains reared in our laboratory and alcohol samples of 10 other P. persimilis strains for the occurrence of Wolbachia by PCR, none of which tested positive. Synthesis of our study and previous reports suggests that infection of P. persimilis with Wolbachia is extremely rare and of minor importance. We discuss the significance of our findings for future studies on the presence of Wolbachia in predaceous arthropods.

Wolbachia transmission dynamics in Formica wood ants

PubMed Central

2008-01-01

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

Wolbachia endosymbionts in haplodiploid and diploid scolytine beetles (Coleoptera: Curculionidae: Scolytinae).

PubMed

Kawasaki, Yuuki; Schuler, Hannes; Stauffer, Christian; Lakatos, Ferenc; Kajimura, Hisashi

2016-05-19

Haplodiploidy is a sex determination system in which fertilized diploid eggs develop into females and unfertilized haploid eggs develop into males. The evolutionary explanations for this phenomenon include the possibility that haplodiploidy can be reinforced by infection with endosymbiotic bacteria, such as Wolbachia. The subfamily Scolytinae contains species with haplodiploid and diploid sex determination systems. Thus, we studied the association with Wolbachia in 12 diploid and 11 haplodiploid scolytine beetles by analyzing wsp and multilocus sequence typing (MLST) of five loci in this endosymbiont. Wolbachia genotypes were compared with mitochondrial (COI) and nuclear (EF) genotypes in the scolytines. Eight of the 23 scolytine species were infected with Wolbachia, with haplodiploids at significantly higher rates than diploid species. Cloning and sequencing detected multiple infections with up to six Wolbachia strains in individual species. Phylogenetic analyses of wsp and five MLST genes revealed different Wolbachia strains in scolytines. Comparisons between the beetle and Wolbachia phylogenies revealed that closely related beetles were infected with genetically different Wolbachia strains. These results suggest the horizontal transmission of multiple Wolbachia strains between scolytines. We discuss these results in terms of the evolution of different sex determination systems in scolytine beetles. © 2016 Society for Applied Microbiology and John Wiley & Sons Ltd.

Does a parthenogenesis-inducing Wolbachia induce vestigial cytoplasmic incompatibility?

NASA Astrophysics Data System (ADS)

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

2011-03-01

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

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

USDA-ARS?s Scientific Manuscript database

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

The route of infection determines Wolbachia antibacterial protection in Drosophila.

PubMed

Gupta, Vanika; Vasanthakrishnan, Radhakrishnan B; Siva-Jothy, Jonathon; Monteith, Katy M; Brown, Sam P; Vale, Pedro F

2017-06-14

Bacterial symbionts are widespread among metazoans and provide a range of beneficial functions. Wolbachia -mediated protection against viral infection has been extensively demonstrated in Drosophila. In mosquitoes that are artificially transinfected with Drosophila melanogaster Wolbachia (wMel), protection from both viral and bacterial infections has been demonstrated. However, no evidence for Wolbachia -mediated antibacterial protection has been demonstrated in Drosophila to date. Here, we show that the route of infection is key for Wolbachia -mediated antibacterial protection. Drosophila melanogaster carrying Wolbachia showed reduced mortality during enteric-but not systemic-infection with the opportunist pathogen Pseudomonas aeruginosa Wolbachia -mediated protection was more pronounced in male flies and is associated with increased early expression of the antimicrobial peptide Attacin A , and also increased expression of a reactive oxygen species detoxification gene ( Gst D8 ). These results highlight that the route of infection is important for symbiont-mediated protection from infection, that Wolbachia can protect hosts by eliciting a combination of resistance and disease tolerance mechanisms, and that these effects are sexually dimorphic. We discuss the importance of using ecologically relevant routes of infection to gain a better understanding of symbiont-mediated protection. © 2017 The Authors.

Obligatory symbiotic Wolbachia endobacteria are absent from Loa loa

PubMed Central

Büttner, Dietrich W; Wanji, Samuel; Bazzocchi, Chiara; Bain, Odile; Fischer, Peter

2003-01-01

Background Many filarial nematodes harbour Wolbachia endobacteria. These endobacteria are transmitted vertically from one generation to the next. In several filarial species that have been studied to date they are obligatory symbionts of their hosts. Elimination of the endobacteria by antibiotics interrupts the embryogenesis and hence the production of microfilariae. The medical implication of this being that the use of doxycycline for the treatment of human onchocerciasis and bancroftian filariasis leads to elimination of the Wolbachia and hence sterilisation of the female worms. Wolbachia play a role in the immunopathology of patients and may contribute to side effects seen after antifilarial chemotherapy. In several studies Wolbachia were not observed in Loa loa. Since these results have been doubted, and because of the medical significance, several independent methods were applied to search for Wolbachia in L. loa. Methods Loa loa and Onchocerca volvulus were studied by electron microscopy, histology with silver staining, and immunohistology using antibodies against WSP, Wolbachia aspartate aminotransferase, and heat shock protein 60. The results achieved with L. loa and O. volvulus were compared. Searching for Wolbachia, genes were amplified by PCR coding for the bacterial 16S rDNA, the FTSZ cell division protein, and WSP. Results No Wolbachia endobacteria were discovered by immunohistology in 13 male and 14 female L. loa worms and in numerous L. loa microfilariae. In contrast, endobacteria were found in large numbers in O. volvulus and 14 other filaria species. No intracellular bacteria were seen in electron micrographs of oocytes and young morulae of L. loa in contrast to O. volvulus. In agreement with these results, Wolbachia DNA was not detected by PCR in three male and six female L. loa worms and in two microfilariae samples of L. loa. Conclusions Loa loa do not harbour obligatory symbiotic Wolbachia endobacteria in essential numbers to enable their

The Endosymbiotic Bacterium Wolbachia Induces Resistance to Dengue Virus in Aedes aegypti

PubMed Central

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

2010-01-01

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

Wolbachia infection complexity among insects in the tropical rice-field community.

PubMed

Kittayapong, P; Jamnongluk, W; Thipaksorn, A; Milne, J R; Sindhusake, C

2003-04-01

Wolbachia are a group of intracellular bacteria that cause reproductive alterations in their arthropod hosts. Widely discordant host and Wolbachia phylogenies indicate that horizontal transmission of these bacteria among species sometimes occurs. A likely means of horizontal transfer is through the feeding relations of organisms within communities. Feeding interactions among insects within the rice-field insect community have been well documented in the past. Here, we present the results of a polymerase chain reaction-based survey and phylogenetic analysis of Wolbachia strains in the rice-field insect community of Thailand. Our field survey indicated that 49 of 209 (23.4%) rice-field insect species were infected with Wolbachia. Of the 49 infected species, 27 were members of two feeding complexes: (i) a group of 13 hoppers preyed on by 2 mirid species and parasitized by a fly species, and (ii) 2 lepidopteran pests parasitized by 9 wasp species. Wolbachia strains found in three hoppers, Recilia dorsalis, Nephotettix malayanus and Nisia nervosa, the two mirid predators, Cyrtorhinus lividipennis and Tytthus chinensis, and the fly parasitoid, Tomosvaryella subvirescens, were all in the same Wolbachia clade. In the second complex, the two lepidopteran pests, Cnaphalocrocis medinalis and Scirpophaga incertulas, were both infected with Wolbachia from the same clade, as was the parasitoid Tropobracon schoenobii. However, none of the other infected parasitoid species in this feeding complex was infected by Wolbachia from this clade. Mean (+/- SD) genetic distance of Wolbachia wsp sequences among interacting species pairs of the hopper feeding complex (0.118 +/- 0.091 nucleotide sequence differences), but not for the other two complexes, was significantly smaller than that between noninteracting species pairs (0.162 +/- 0.079 nucleotide sequence differences). Our results suggest that some feeding complexes, such as the hopper complex described here, could be an important

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

PubMed

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

2016-10-13

Dactylopius species, known as cochineal insects, are the source of the carminic acid dye used worldwide. The presence of two Wolbachia strains in Dactylopius coccus from Mexico was revealed by PCR amplification of wsp and sequencing of 16S rRNA genes. A metagenome analysis recovered the genome sequences of Candidatus Wolbachia bourtzisii wDacA (supergroup A) and Candidatus Wolbachia pipientis wDacB (supergroup B). Genome read coverage, as well as 16S rRNA clone sequencing, revealed that wDacB was more abundant than wDacA. The strains shared similar predicted metabolic capabilities that are common to Wolbachia, including riboflavin, ubiquinone, and heme biosynthesis, but lacked other vitamin and cofactor biosynthesis as well as glycolysis, the oxidative pentose phosphate pathway, and sugar uptake systems. A complete tricarboxylic acid cycle and gluconeogenesis were predicted as well as limited amino acid biosynthesis. Uptake and catabolism of proline were evidenced in Dactylopius Wolbachia strains. Both strains possessed WO-like phage regions and type I and type IV secretion systems. Several efflux systems found suggested the existence of metal toxicity within their host. Besides already described putative virulence factors like ankyrin domain proteins, VlrC homologs, and patatin-like proteins, putative novel virulence factors related to those found in intracellular pathogens like Legionella and Mycobacterium are highlighted for the first time in Wolbachia Candidate genes identified in other Wolbachia that are likely involved in cytoplasmic incompatibility were found in wDacB but not in wDacA. Copyright © 2016 Ramírez-Puebla et al.

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

Wolbachia infections in native and introduced populations of fire ants (Solenopsis spp.).

PubMed

Shoemaker, D D; Ross, K G; Keller, L; Vargo, E L; Werren, J H

2000-12-01

Wolbachia are cytoplasmically inherited bacteria that induce a variety of effects with fitness consequences on host arthropods, including cytoplasmic incompatibility, parthenogenesis, male-killing and feminization. We report here the presence of Wolbachia in native South American populations of the fire ant Solenopsis invicta, but the apparent absence of the bacteria in introduced populations of this pest species in the USA. The Wolbachia strains in native S. invicta are of two divergent types (A and B), and the frequency of infection varies dramatically between geographical regions and social forms of this host. Survey data reveal that Wolbachia also are found in other native fire ant species within the Solenopsis saevissima species complex from South America, including S. richteri. This latter species also has been introduced in the USA, where it lacks Wolbachia. Sequence data reveal complete phylogenetic concordance between mtDNA haplotype in S. invicta and Wolbachia infection type (A or B). In addition, the mtDNA and associated group A Wolbachia strain in S. invicta are more closely related to the mtDNA and Wolbachia strain found in S. richteri than they are to the mtDNA and associated group B Wolbachia in S. invicta. These data are consistent with historical introgression of S. richteri cytoplasmic elements into S. invicta populations, resulting in enhanced infection and mtDNA polymorphisms in S. invicta. Wolbachia may have significant fitness effects on these hosts (either directly or by cytoplasmic incompatibility) and therefore these microbes potentially could be used in biological control programmes to suppress introduced fire ant populations.

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

PubMed

Watanabe, Masaya; Kageyama, Daisuke; Miura, Kazuki

2013-01-01

Wolbachia, which are maternally transmitted endosymbionts, are considered to have moved horizontally between invertebrate hosts multiple times. However, it is not well understood how easily Wolbachia are transmitted horizontally between different hosts and how frequently horizontally-transmitted Wolbachia become established in their new hosts. We transferred a parthenogenesis-inducing Wolbachia endosymbiont derived from the parasitic wasp Trichogramma dendrolimi to Trichogramma evanescens. Specifically, Wolbachia was cultivated in a mosquito cell line and the Wolbachia-infected cells were microinjected into uninfected T. evanescens. Among 276 pupae inoculated with Wolbachia-infected cells, 65 adults emerged (G0). Diagnostic PCR demonstrated that 25 of 37 G0 females (68%) were Wolbachia-positive. Among isofemale lines established from G0 females, the proportions of infected lines were 80% (20 of 25) in G1 and 100% (18 of 18) in G2. In an isofemale line, infection was stably maintained for more than 10 generations. These results indicate invasion of Wolbachia into the germline of the recipient insect. Quantitative PCR demonstrated that the Wolbachia titer in the recipient host was significantly lower than that in the native host. The absence or very low number, if any, of parthenogenetically-reproducing individuals in the recipient host may be caused by the low Wolbachia titer. The Wolbachia titer in the recipients was lower in G11 than in G5, suggesting a decline in the density. Together with a previous report, our study may imply that Wolbachia in Trichogramma species are highly adapted to their hosts, which hinders robust expression of the Wolbachia phenotype in non-native host species. Copyright © 2012 Elsevier Inc. All rights reserved.

Classification of endosymbiont Wolbachia (Rickettsiales: Anaplasmataceae) in opiine wasps (Hymenoptera: Braconidae)

NASA Astrophysics Data System (ADS)

Mohammed, Muhamad Azmi; Zuki, Ameyra Aman; Yusof, Suhana; Othman, Nurul Wahida; Zain, Badrul Munir Md; Yaakop, Salmah

2018-04-01

Endosymbiont Wolbachia has always been a hot topic of discussion among entomologists and microbiologists as it can manipulate the reproductive system of their arthropod hosts. In this study, a total of 10 sequences which consist of concatenate data from three genetic markers of Wolbachia (groEL, gltA, and wsp) were obtained from opiine wasps from five localities in Peninsular Malaysia. Among the 10 sequences, six were isolated from Fopius arisanus, one from F. vandenboschi, and three from Psyttalia sp. SY2013. Based on Neighbour-Joining (NJ) analysis of the concatenate data and genetic distances, four variants of Wolbachia have been successfully identified. Our data thus provide an insight on Wolbachia infections in oriental insects as Wolbachia research is still considered as in early stage in Malaysia.

Distribution and phylogeny of Wolbachia strains in wild mosquito populations in Sri Lanka.

PubMed

Nugapola, N W Nalaka P; De Silva, W A Priyanka P; Karunaratne, S H P Parakrama

2017-05-10

Wolbachia are a group of maternally inherited intracellular bacteria known to be widespread among arthropods. Infections with Wolbachia cause declines of host populations, and also induce host resistance to a wide range of pathogens. Over the past few decades, researchers were curious to use Wolbachia as a biological tool to control mosquito vectors. During the present study, assessment of the prevalence of Wolbachia infections among wild mosquito populations in Sri Lanka where mosquito-borne diseases are a major health concern, was carried out for the first time. DNA was extracted from the abdomens of mosquitoes, collected from seven provinces, and screened for the presence of Wolbachia by PCR using wsp and groE primers. Group-specific and strain-specific primers were used to classify Wolbachia into the supergroups A and B, and into the strains Mel, AlbA and Pip. A total of 330 individual mosquitoes belonging to 22 species and 7 genera were screened. Eighty-seven mosquitoes (26.36%) belonging to four species (i.e. Aedes albopictus, Culex quinquefasciatus, Armigeres subalbatus and Mansonia uniformis) were positive for Wolbachia infections. Primary vector of the dengue fever, Ae. aegypti was negative for Wolbachia infections while the secondary vector, Ae. albopictus, showed a very high infection rate. The filarial vector C. quinquefasciatus had a relatively high rate of infection. Japanese encephalitis vectors C. gelidus and C. triteaneorynchus, and the Anopheles vectors of malaria were negative for Wolbachia infections. Nine sequences of Wolbachia-positive PCR products were deposited in the GenBank and compared with other available data. Aedes albopictus was infected with both Wolbachia strains A (AlbA) and B (Pip) supergroups. Phylogenetic analysis of the wsp sequences showed two major branches confirming identities obtained from the PCR screening with strain-specific primers. Wolbachia infections were found only among four mosquito species in Sri Lanka: Aedes

Association of Wolbachia with heartworm disease in cats and dogs.

PubMed

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

2010-05-28

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

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

PubMed Central

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

2016-01-01

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

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

PubMed

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

2004-10-19

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.

A critical re-evaluation of multilocus sequence typing (MLST) efforts in Wolbachia.

PubMed

Bleidorn, Christoph; Gerth, Michael

2018-01-01

Wolbachia (Alphaproteobacteria, Rickettsiales) is the most common, and arguably one of the most important inherited symbionts. Molecular differentiation of Wolbachia strains is routinely performed with a set of five multilocus sequence typing (MLST) markers. However, since its inception in 2006, the performance of MLST in Wolbachia strain typing has not been assessed objectively. Here, we evaluate the properties of Wolbachia MLST markers and compare it to 252 other single copy loci present in the genome of most Wolbachia strains. Specifically, we investigated how well MLST performs at strain differentiation, at reflecting genetic diversity of strains, and as phylogenetic marker. We find that MLST loci are outperformed by other loci at all tasks they are currently employed for, and thus that they do not reflect the properties of a Wolbachia strain very well. We argue that whole genome typing approaches should be used for Wolbachia typing in the future. Alternatively, if few loci approaches are necessary, we provide a characterisation of 252 single copy loci for a number a criteria, which may assist in designing specific typing systems or phylogenetic studies. © FEMS 2017. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

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

PubMed Central

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

2016-01-01

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

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

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

PubMed Central

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

2017-01-01

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

The Gut Commensal Microbiome of Drosophila melanogaster Is Modified by the Endosymbiont Wolbachia

PubMed Central

Fast, Eva M.; Guo, Rong; Vaisman, Natalie; Ortiz, Luis; Bybee, Joanna; Slatko, Barton E.

2017-01-01

ABSTRACT Endosymbiotic Wolbachia bacteria and the gut microbiome have independently been shown to affect several aspects of insect biology, including reproduction, development, life span, stem cell activity, and resistance to human pathogens, in insect vectors. This work shows that Wolbachia bacteria, which reside mainly in the fly germline, affect the microbial species present in the fly gut in a lab-reared strain. Drosophila melanogaster hosts two main genera of commensal bacteria—Acetobacter and Lactobacillus. Wolbachia-infected flies have significantly reduced titers of Acetobacter. Sampling of the microbiome of axenic flies fed with equal proportions of both bacteria shows that the presence of Wolbachia bacteria is a significant determinant of the composition of the microbiome throughout fly development. However, this effect is host genotype dependent. To investigate the mechanism of microbiome modulation, the effect of Wolbachia bacteria on Imd and reactive oxygen species pathways, the main regulators of immune response in the fly gut, was measured. The presence of Wolbachia bacteria does not induce significant changes in the expression of the genes for the effector molecules in either pathway. Furthermore, microbiome modulation is not due to direct interaction between Wolbachia bacteria and gut microbes. Confocal analysis shows that Wolbachia bacteria are absent from the gut lumen. These results indicate that the mechanistic basis of the modulation of composition of the microbiome by Wolbachia bacteria is more complex than a direct bacterial interaction or the effect of Wolbachia bacteria on fly immunity. The findings reported here highlight the importance of considering the composition of the gut microbiome and host genetic background during Wolbachia-induced phenotypic studies and when formulating microbe-based disease vector control strategies. IMPORTANCE Wolbachia bacteria are intracellular bacteria present in the microbiome of a large fraction of

The Gut Commensal Microbiome of Drosophila melanogaster Is Modified by the Endosymbiont Wolbachia.

PubMed

Simhadri, Rama K; Fast, Eva M; Guo, Rong; Schultz, Michaela J; Vaisman, Natalie; Ortiz, Luis; Bybee, Joanna; Slatko, Barton E; Frydman, Horacio M

2017-01-01

Endosymbiotic Wolbachia bacteria and the gut microbiome have independently been shown to affect several aspects of insect biology, including reproduction, development, life span, stem cell activity, and resistance to human pathogens, in insect vectors. This work shows that Wolbachia bacteria, which reside mainly in the fly germline, affect the microbial species present in the fly gut in a lab-reared strain. Drosophila melanogaster hosts two main genera of commensal bacteria- Acetobacter and Lactobacillus . Wolbachia -infected flies have significantly reduced titers of Acetobacter . Sampling of the microbiome of axenic flies fed with equal proportions of both bacteria shows that the presence of Wolbachia bacteria is a significant determinant of the composition of the microbiome throughout fly development. However, this effect is host genotype dependent. To investigate the mechanism of microbiome modulation, the effect of Wolbachia bacteria on Imd and reactive oxygen species pathways, the main regulators of immune response in the fly gut, was measured. The presence of Wolbachia bacteria does not induce significant changes in the expression of the genes for the effector molecules in either pathway. Furthermore, microbiome modulation is not due to direct interaction between Wolbachia bacteria and gut microbes. Confocal analysis shows that Wolbachia bacteria are absent from the gut lumen. These results indicate that the mechanistic basis of the modulation of composition of the microbiome by Wolbachia bacteria is more complex than a direct bacterial interaction or the effect of Wolbachia bacteria on fly immunity. The findings reported here highlight the importance of considering the composition of the gut microbiome and host genetic background during Wolbachia -induced phenotypic studies and when formulating microbe-based disease vector control strategies. IMPORTANCE Wolbachia bacteria are intracellular bacteria present in the microbiome of a large fraction of insects

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

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

PubMed

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

2015-06-01

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

Modelling Wolbachia infection in a sex-structured mosquito population carrying West Nile virus.

PubMed

Farkas, József Z; Gourley, Stephen A; Liu, Rongsong; Yakubu, Abdul-Aziz

2017-09-01

Wolbachia is possibly the most studied reproductive parasite of arthropod species. It appears to be a promising candidate for biocontrol of some mosquito borne diseases. We begin by developing a sex-structured model for a Wolbachia infected mosquito population. Our model incorporates the key effects of Wolbachia infection including cytoplasmic incompatibility and male killing. We also allow the possibility of reduced reproductive output, incomplete maternal transmission, and different mortality rates for uninfected/infected male/female individuals. We study the existence and local stability of equilibria, including the biologically relevant and interesting boundary equilibria. For some biologically relevant parameter regimes there may be multiple coexistence steady states including, very importantly, a coexistence steady state in which Wolbachia infected individuals dominate. We also extend the model to incorporate West Nile virus (WNv) dynamics, using an SEI modelling approach. Recent evidence suggests that a particular strain of Wolbachia infection significantly reduces WNv replication in Aedes aegypti. We model this via increased time spent in the WNv-exposed compartment for Wolbachia infected female mosquitoes. A basic reproduction number [Formula: see text] is computed for the WNv infection. Our results suggest that, if the mosquito population consists mainly of Wolbachia infected individuals, WNv eradication is likely if WNv replication in Wolbachia infected individuals is sufficiently reduced.

Wolbachia influences the maternal transmission of the gypsy endogenous retrovirus in Drosophila melanogaster.

PubMed

Touret, Franck; Guiguen, François; Terzian, Christophe

2014-09-02

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. Animals have established complex relationships with bacteria and viruses that spread horizontally among individuals or are vertically transmitted, i.e., from parents to offspring. It is well established that members of the genus Wolbachia, maternally inherited symbiotic bacteria present mainly in arthropods, reduce the replication of several RNA viruses transmitted horizontally. Here, we demonstrate for the first time that Wolbachia diminishes the maternal transmission of gypsy, an endogenous retrovirus in Drosophila melanogaster. We hypothesize that gypsy cannot efficiently integrate into the germ

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

PubMed Central

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

2010-01-01

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

Feminizing Wolbachia influence microbiota composition in the terrestrial isopod Armadillidium vulgare.

PubMed

Dittmer, Jessica; Bouchon, Didier

2018-05-03

Wolbachia are widespread heritable endosymbionts of arthropods notorious for their profound effects on host fitness as well as for providing protection against viruses and eukaryotic parasites, indicating that they can interact with other microorganisms sharing the same host environment. Using the terrestrial isopod crustacean Armadillidium vulgare, its highly diverse microbiota (>200 bacterial genera) and its three feminizing Wolbachia strains (wVulC, wVulM, wVulP) as a model system, the present study demonstrates that Wolbachia can even influence the composition of a diverse bacterial community under both laboratory and natural conditions. While host origin is the major determinant of the taxonomic composition of the microbiota in A. vulgare, Wolbachia infection affected both the presence and, more importantly, the abundance of many bacterial taxa within each host population, possibly due to competitive interactions. Moreover, different Wolbachia strains had different impacts on microbiota composition. As such, infection with wVulC affected a higher number of taxa than infection with wVulM, possibly due to intrinsic differences in virulence and titer between these two strains. In conclusion, this study shows that heritable endosymbionts such as Wolbachia can act as biotic factors shaping the microbiota of arthropods, with as yet unknown consequences on host fitness.

Wolbachia density changes seasonally amongst populations of the pale grass blue butterfly, Zizeeria maha (Lepidoptera: Lycaenidae)

PubMed Central

Sumi, Takuto; Miura, Kazuki; Miyatake, Takahisa

2017-01-01

Previous studies showed that the survival rate of Wolbachia decreases under high temperature in incubators. It is also known that a high density of Wolbachia in the host body reduces the host emergence rate, while low densities fail to change reproduction rates. However, few studies have examined the density of Wolbachia in hosts in the field. Here, we focus on Wolbachia infection of the pale grass blue butterfly, Zizeeria maha (Lepidoptera: Lycaenidae), which is distributed throughout the Japanese islands. We examined the rate and density of Wolbachia infection in the bodies of butterflies at thirteen locations in Japan. At seven of these places, we collected butterflies in different seasons to determine seasonal differences in the infection rate and density and found that Wolbachia density has seasonal differences within the same population. Moreover, to determine whether Wolbachia density has a geographical cline, we compared the infection density of Wolbachia amongst all geographical populations. In addition, we determined the sequences of Wolbachia wsp and host mtDNA CO1 haplotypes of all populations. The results showed that Wolbachia density increased in early summer and decreased in autumn. Further, the density of Wolbachia infecting the same strain of Z. maha varied amongst populations, although no tendency in geographical cline was observed. PMID:28403227

Wolbachia density changes seasonally amongst populations of the pale grass blue butterfly, Zizeeria maha (Lepidoptera: Lycaenidae).

PubMed

Sumi, Takuto; Miura, Kazuki; Miyatake, Takahisa

2017-01-01

Previous studies showed that the survival rate of Wolbachia decreases under high temperature in incubators. It is also known that a high density of Wolbachia in the host body reduces the host emergence rate, while low densities fail to change reproduction rates. However, few studies have examined the density of Wolbachia in hosts in the field. Here, we focus on Wolbachia infection of the pale grass blue butterfly, Zizeeria maha (Lepidoptera: Lycaenidae), which is distributed throughout the Japanese islands. We examined the rate and density of Wolbachia infection in the bodies of butterflies at thirteen locations in Japan. At seven of these places, we collected butterflies in different seasons to determine seasonal differences in the infection rate and density and found that Wolbachia density has seasonal differences within the same population. Moreover, to determine whether Wolbachia density has a geographical cline, we compared the infection density of Wolbachia amongst all geographical populations. In addition, we determined the sequences of Wolbachia wsp and host mtDNA CO1 haplotypes of all populations. The results showed that Wolbachia density increased in early summer and decreased in autumn. Further, the density of Wolbachia infecting the same strain of Z. maha varied amongst populations, although no tendency in geographical cline was observed.

Cytonuclear Epistasis Controls the Density of Symbiont Wolbachia pipientis in Nongonadal Tissues of Mosquito Culex quinquefasciatus.

PubMed

Emerson, Kevin J; Glaser, Robert L

2017-08-07

Wolbachia pipientis , a bacterial symbiont infecting arthropods and nematodes, is vertically transmitted through the female germline and manipulates its host's reproduction to favor infected females. Wolbachia also infects somatic tissues where it can cause nonreproductive phenotypes in its host, including resistance to viral pathogens. Wolbachia -mediated phenotypes are strongly associated with the density of Wolbachia in host tissues. Little is known, however, about how Wolbachia density is regulated in native or heterologous hosts. Here, we measure the broad-sense heritability of Wolbachia density among families in field populations of the mosquito Culex pipiens , and show that densities in ovary and nongonadal tissues of females in the same family are not correlated, suggesting that Wolbachia density is determined by distinct mechanisms in the two tissues. Using introgression analysis between two different strains of the closely related species C. quinquefasciatus , we show that Wolbachia densities in ovary tissues are determined primarily by cytoplasmic genotype, while densities in nongonadal tissues are determined by both cytoplasmic and nuclear genotypes and their epistatic interactions. Quantitative-trait-locus mapping identified two major-effect quantitative-trait loci in the C. quinquefasciatus genome explaining a combined 23% of variance in Wolbachia density, specifically in nongonadal tissues. A better understanding of how Wolbachia density is regulated will provide insights into how Wolbachia density can vary spatiotemporally in insect populations, leading to changes in Wolbachia -mediated phenotypes such as viral pathogen resistance. Copyright © 2017 Emerson, Glaser.

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

PubMed Central

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

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

PubMed

Joubert, Dirk Albert; O'Neill, Scott L

2017-01-01

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

Current state of knowledge on Wolbachia infection among Coleoptera: a systematic review

PubMed Central

Kotásková, Nela

2018-01-01

Background Despite great progress in studies on Wolbachia infection in insects, the knowledge about its relations with beetle species, populations and individuals, and the effects of bacteria on these hosts, is still unsatisfactory. In this review we summarize the current state of knowledge about Wolbachia occurrence and interactions with Coleopteran hosts. Methods An intensive search of the available literature resulted in the selection of 86 publications that describe the relevant details about Wolbachia presence among beetles. These publications were then examined with respect to the distribution and taxonomy of infected hosts and diversity of Wolbachia found in beetles. Sequences of Wolbachia genes (16S rDNA, ftsZ) were used for the phylogenetic analyses. Results The collected publications revealed that Wolbachia has been confirmed in 204 beetle species and that the estimated average prevalence of this bacteria across beetle species is 38.3% and varies greatly across families and genera (0–88% infected members) and is much lower (c. 13%) in geographic studies. The majority of the examined and infected beetles were from Europe and East Asia. The most intensively studied have been two groups of herbivorous beetles: Curculionidae and Chrysomelidae. Coleoptera harbor Wolbachia belonging to three supergroups: F found in only three species, and A and B found in similar numbers of beetles (including some doubly infected); however the latter two were most prevalent in different families. A total of 59% of species with precise data were found to be totally infected. Single infections were found in 69% of species and others were doubly- or multiply-infected. Wolbachia caused numerous effects on its beetle hosts, including selective sweep with host mtDNA (found in 3% of species), cytoplasmic incompatibility (detected in c. 6% of beetles) and other effects related to reproduction or development (like male-killing, possible parthenogenesis or haplodiploidy induction, and

The effects of outbreeding on a parasitoid wasp fixed for infection with a parthenogenesis-inducing Wolbachia symbiont.

PubMed

Lindsey, A R I; Stouthamer, R

2017-12-01

Trichogramma wasps can be rendered asexual by infection with the maternally inherited symbiont Wolbachia. Previous studies indicate the Wolbachia strains infecting Trichogramma wasps are host-specific, inferred by failed horizontal transfer of Wolbachia to novel Trichogramma hosts. Additionally, Trichogramma can become dependent upon their Wolbachia infection for the production of female offspring, leaving them irreversibly asexual, further linking host and symbiont. We hypothesized Wolbachia strains infecting irreversibly asexual, resistant to horizontal transfer Trichogramma would show adaptation to a particular host genetic background. To test this, we mated Wolbachia-dependent females with males from a Wolbachia-naïve population to create heterozygous wasps. We measured sex ratios and fecundity, a proxy for Wolbachia fitness, produced by heterozygous wasps, and by their recombinant offspring. We find a heterozygote advantage, resulting in higher fitness for Wolbachia, as wasps will produce more offspring without any reduction in the proportion of females. While recombinant wasps did not differ in total fecundity after 10 days, recombinants produced fewer offspring early on, leading to an increased female-biased sex ratio for the whole brood. Despite the previously identified barriers to horizontal transfer of Wolbachia to and from Trichogramma pretiosum, there were no apparent barriers for Wolbachia to induce parthenogenesis in these non-native backgrounds. This is likely due to the route of infection being introgression rather than horizontal transfer, and possibly the co-evolution of Wolbachia with the mitochondria rather than the nuclear genome. These results help to elucidate the mechanisms by which Wolbachia adapt to hosts and the evolution of host-symbiont phenotypes.

Detection of Wolbachia endobacteria in Culex quinquefasciatus by Gimenez staining and confirmation by PCR.

PubMed

Muniaraj, M; Paramasivan, R; Sunish, I P; Arunachalam, N; Mariappan, T; Jerald Leo, S Victor; Dhananjeyan, K J

2012-12-01

Wolbachia are common intracellular bacteria that are found in arthropods and nematodes. These endosymbionts are transmitted vertically through host eggs and alter host biology in diverse ways, including the induction of reproductive manipulations, such as feminization, parthenogenesis, male killing and sperm-egg incompatibility. Since they can also move horizontally across species boundaries, Wolbachia is gaining importance in recent days as it could be used as a biological control agent to control vector mosquitoes or for paratransgenic approaches. However, the study of Wolbachia requires sophisticated techniques such as PCR and cell culture facilities which cannot be affordable for many laboratories where the diseases transmitted by arthropod vectors are common. Hence, it would be beneficial to develop a simple method to detect the presence of Wolbachia in arthropods. In this study, we described a method of staining Wolbachia endobacteria, present in the reproductive tissues of mosquitoes. The reliability of this method was compared with Gram staining and PCR based detection. The microscopic observation of the Gimenez stained smear prepared from the teased ovary of wild caught and Wolbachia (+) Cx. quinquefasciatus revealed the presence of pink coloured pleomorphic cells of Wolbachia ranging from cocci, comma shaped cells to bacillus and chain forms. The ovaries of Wolbachia (-) cured mosquito did not show any cell. Although Gram's staining is a reliable differential staining for the other bacteria, the bacterial cells in the smears from the ovaries of wild caught mosquitoes did not take the stain properly and the cells were not clearly visible. The PCR amplified product from the pooled remains of wild caught and Wolbachia (+) Cx. quinquefasciatus showed clear banding, whereas, no banding was observed for the negative control (distilled water) and Wolbachia (-) Cx. quinquefasciatus. The Gimenez staining technique applied, could be used to detect the members of

Genome-wide analysis of the interaction between the endosymbiotic bacterium Wolbachia and its Drosophila host.

PubMed

Xi, Zhiyong; Gavotte, Laurent; Xie, Yan; Dobson, Stephen L

2008-01-02

Intracellular Wolbachia bacteria are obligate, maternally-inherited, endosymbionts found frequently in insects and other invertebrates. The success of Wolbachia can be attributed in part to an ability to alter host reproduction via mechanisms including cytoplasmic incompatibility (CI), parthenogenesis, feminization and male killing. Despite substantial scientific effort, the molecular mechanisms underlying the Wolbachia/host interaction are unknown. Here, an in vitro Wolbachia infection was generated in the Drosophila S2 cell line, and transcription profiles of infected and uninfected cells were compared by microarray. Differentially-expressed patterns related to reproduction, immune response and heat stress response are observed, including multiple genes that have been previously reported to be involved in the Wolbachia/host interaction. Subsequent in vivo characterization of differentially-expressed products in gonads demonstrates that Angiotensin Converting Enzyme (Ance) varies between Wolbachia infected and uninfected flies and that the variation occurs in a sex-specific manner. Consistent with expectations for the conserved CI mechanism, the observed Ance expression pattern is repeatable in different Drosophila species and with different Wolbachia types. To examine Ance involvement in the CI phenotype, compatible and incompatible crosses of Ance mutant flies were conducted. Significant differences are observed in the egg hatch rate resulting from incompatible crosses, providing support for additional experiments examining for an interaction of Ance with the CI mechanism. Wolbachia infection is shown to affect the expression of multiple host genes, including Ance. Evidence for potential Ance involvement in the CI mechanism is described, including the prior report of Ance in spermatid differentiation, Wolbachia-induced sex-specific effects on Ance expression and an Ance mutation effect on CI levels. The results support the use of Wolbachia infected cell cultures

Genome-wide analysis of the interaction between the endosymbiotic bacterium Wolbachia and its Drosophila host

PubMed Central

Xi, Zhiyong; Gavotte, Laurent; Xie, Yan; Dobson, Stephen L

2008-01-01

Background Intracellular Wolbachia bacteria are obligate, maternally-inherited, endosymbionts found frequently in insects and other invertebrates. The success of Wolbachia can be attributed in part to an ability to alter host reproduction via mechanisms including cytoplasmic incompatibility (CI), parthenogenesis, feminization and male killing. Despite substantial scientific effort, the molecular mechanisms underlying the Wolbachia/host interaction are unknown. Results Here, an in vitro Wolbachia infection was generated in the Drosophila S2 cell line, and transcription profiles of infected and uninfected cells were compared by microarray. Differentially-expressed patterns related to reproduction, immune response and heat stress response are observed, including multiple genes that have been previously reported to be involved in the Wolbachia/host interaction. Subsequent in vivo characterization of differentially-expressed products in gonads demonstrates that Angiotensin Converting Enzyme (Ance) varies between Wolbachia infected and uninfected flies and that the variation occurs in a sex-specific manner. Consistent with expectations for the conserved CI mechanism, the observed Ance expression pattern is repeatable in different Drosophila species and with different Wolbachia types. To examine Ance involvement in the CI phenotype, compatible and incompatible crosses of Ance mutant flies were conducted. Significant differences are observed in the egg hatch rate resulting from incompatible crosses, providing support for additional experiments examining for an interaction of Ance with the CI mechanism. Conclusion Wolbachia infection is shown to affect the expression of multiple host genes, including Ance. Evidence for potential Ance involvement in the CI mechanism is described, including the prior report of Ance in spermatid differentiation, Wolbachia-induced sex-specific effects on Ance expression and an Ance mutation effect on CI levels. The results support the use of

Wolbachia infection reduces blood-feeding success in the dengue fever mosquito, Aedes aegypti.

PubMed

Turley, Andrew P; Moreira, Luciano A; O'Neill, Scott L; McGraw, Elizabeth A

2009-09-15

The mosquito Aedes aegypti was recently transinfected with a life-shortening strain of the endosymbiont Wolbachia pipientis (wMelPop) as the first step in developing a biocontrol strategy for dengue virus transmission. In addition to life-shortening, the wMelPop-infected mosquitoes also exhibit increased daytime activity and metabolic rates. Here we sought to quantify the blood-feeding behaviour of Wolbachia-infected females as an indicator of any virulence or energetic drain associated with Wolbachia infection. In a series of blood-feeding trials in response to humans, we have shown that Wolbachia-infected mosquitoes do not differ in their response time to humans, but that as they age they obtain fewer and smaller blood meals than Wolbachia-uninfected controls. Lastly, we observed a behavioural characteristic in the Wolbachia infected mosquitoes best described as a "bendy" proboscis that may explain the decreased biting success. Taken together the evidence suggests that wMelPop infection may be causing tissue damage in a manner that intensifies with mosquito age and that leads to reduced blood-feeding success. These behavioural changes require further investigation with respect to a possible physiological mechanism and their role in vectorial capacity of the insect. The selective decrease of feeding success in older mosquitoes may act synergistically with other Wolbachia-associated traits including life-shortening and viral protection in biocontrol strategies.

Comprehensive proteome profiling in Aedes albopictus to decipher Wolbachia-arbovirus interference phenomenon.

PubMed

Saucereau, Yoann; Valiente Moro, Claire; Dieryckx, Cindy; Dupuy, Jean-William; Tran, Florence-Hélène; Girard, Vincent; Potier, Patrick; Mavingui, Patrick

2017-08-18

Aedes albopictus is a vector of arboviruses that cause severe diseases in humans such as Chikungunya, Dengue and Zika fevers. The vector competence of Ae. albopictus varies depending on the mosquito population involved and the virus transmitted. Wolbachia infection status in believed to be among key elements that determine viral transmission efficiency. Little is known about the cellular functions mobilized in Ae. albopictus during co-infection by Wolbachia and a given arbovirus. To decipher this tripartite interaction at the molecular level, we performed a proteome analysis in Ae. albopictus C6/36 cells mono-infected by Wolbachia wAlbB strain or Chikungunya virus (CHIKV), and bi-infected. We first confirmed significant inhibition of CHIKV by Wolbachia. Using two-dimensional gel electrophoresis followed by nano liquid chromatography coupled with tandem mass spectrometry, we identified 600 unique differentially expressed proteins mostly related to glycolysis, translation and protein metabolism. Wolbachia infection had greater impact on cellular functions than CHIKV infection, inducing either up or down-regulation of proteins associated with metabolic processes such as glycolysis and ATP metabolism, or structural glycoproteins and capsid proteins in the case of bi-infection with CHIKV. CHIKV infection inhibited expression of proteins linked with the processes of transcription, translation, lipid storage and miRNA pathways. The results of our proteome profiling have provided new insights into the molecular pathways involved in tripartite Ae. albopictus-Wolbachia-CHIKV interaction and may help defining targets for the better implementation of Wolbachia-based strategies for disease transmission control.

Wolbachia: A biological control strategy against arboviral diseases.

PubMed

Mohanty, Ipsita; Rath, Animesha; Mahapatra, Namita; Hazra, Rupenangshu K

2016-01-01

Vector-borne diseases particularly those transmitted by mosquitoes like Dengue are among the leading causes of mortality and morbidity in human population. There are no effective vaccines or treatment against dengue fever till date and the control methods are limited. So, new approaches are urgently in need to reverse these trends. Vector control is currently the primary intervention tool. 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. Wolbachia an endosymbiont of arthropod vectors is being explored as a novel ecofriendly control strategy. Studies in Drosophila have shown that Wolbachia can confer resistance to diverse RNA viruses and protect flies from virus-induced mortality. This review was focused on biology of the Wolbachia and its implication as a control measure for arboviral diseases mainly Dengue and Chikungunya.

Reliance of Wolbachia on High Rates of Host Proteolysis Revealed by a Genome-Wide RNAi Screen of Drosophila Cells.

PubMed

White, Pamela M; Serbus, Laura R; Debec, Alain; Codina, Adan; Bray, Walter; Guichet, Antoine; Lokey, R Scott; Sullivan, William

2017-04-01

Wolbachia are gram-negative, obligate, intracellular bacteria carried by a majority of insect species worldwide. Here we use a Wolbachia -infected Drosophila cell line and genome-wide RNA interference (RNAi) screening to identify host factors that influence Wolbachia titer. By screening an RNAi library targeting 15,699 transcribed host genes, we identified 36 candidate genes that dramatically reduced Wolbachia titer and 41 that increased Wolbachia titer. Host gene knockdowns that reduced Wolbachia titer spanned a broad array of biological pathways including genes that influenced mitochondrial function and lipid metabolism. In addition, knockdown of seven genes in the host ubiquitin and proteolysis pathways significantly reduced Wolbachia titer. To test the in vivo relevance of these results, we found that drug and mutant inhibition of proteolysis reduced levels of Wolbachia in the Drosophila oocyte. The presence of Wolbachia in either cell lines or oocytes dramatically alters the distribution and abundance of ubiquitinated proteins. Functional studies revealed that maintenance of Wolbachia titer relies on an intact host Endoplasmic Reticulum (ER)-associated protein degradation pathway (ERAD). Accordingly, electron microscopy studies demonstrated that Wolbachia is intimately associated with the host ER and dramatically alters the morphology of this organelle. Given Wolbachia lack essential amino acid biosynthetic pathways, the reliance of Wolbachia on high rates of host proteolysis via ubiquitination and the ERAD pathways may be a key mechanism for provisioning Wolbachia with amino acids. In addition, the reliance of Wolbachia on the ERAD pathway and disruption of ER morphology suggests a previously unsuspected mechanism for Wolbachia ' s potent ability to prevent RNA virus replication. Copyright © 2017 by the Genetics Society of America.

A cascade of destabilizations: Combining Wolbachia and Allee effects to eradicate insect pests.

PubMed

Blackwood, Julie C; Vargas, Roger; Fauvergue, Xavier

2018-01-01

The management of insect pests has long been dominated by the use of chemical insecticides, with the aim of instantaneously killing enough individuals to limit their damage. To minimize unwanted consequences, environmentally friendly approaches have been proposed that utilize biological control and take advantage of intrinsic demographic processes to reduce pest populations. We address the feasibility of a novel pest management strategy based on the release of insects infected with Wolbachia, which causes cytoplasmic incompatibilities in its host population, into a population with a pre-existing Allee effect. We hypothesize that the transient decline in population size caused by a successful invasion of Wolbachia can bring the population below its Allee threshold and, consequently, trigger extinction. We develop a stochastic population model that accounts for Wolbachia-induced cytoplasmic incompatibilities in addition to an Allee effect arising from mating failures at low population densities. Using our model, we identify conditions under which cytoplasmic incompatibilities and Allee effects successfully interact to drive insect pest populations towards extinction. Based on our results, we delineate control strategies based on introductions of Wolbachia-infected insects. We extend this analysis to evaluate control strategies that implement successive introductions of two incompatible Wolbachia strains. Additionally, we consider methods that combine Wolbachia invasion with mating disruption tactics to enhance the pre-existing Allee effect. We demonstrate that Wolbachia-induced cytoplasmic incompatibility and the Allee effect act independently from one another: the Allee effect does not modify the Wolbachia invasion threshold, and cytoplasmic incompatibilities only have a marginal effect on the Allee threshold. However, the interaction of these two processes can drive even large populations to extinction. The success of this method can be amplified by the

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

PubMed Central

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

2017-01-01

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

Impacts of temperature and crowding on sex ratio, fecundity and Wolbachia infection intensity in the copepod, Mesocyclops thermocyclopoides.

PubMed

Wiwatanaratanabutr, Itsanun; Grandjean, Frederic

2016-11-01

Wolbachia are a group of intracellular bacteria that cause reproductive alterations in arthropods. Here, we describe the effects of two environmental factors (crowding and temperature) on phenotypic expression of feminization, the host's fecundity and Wolbachia infection intensity among life cycle stages in the naturally Wolbachia-infected copepod, Mesocyclops thermocyclopoides. The copepod was first found to be co-infected with Wolbachia A- and B-supergroups Wolbachia strains based on wsp primers. The relative Wolbachia infection intensity within individuals was determined using quantitative real-time PCR and was significantly higher in the B-supergroup than in the A-supergroup. Experimental results of temperature effect on bacterial density in each developmental stage revealed a significant decrease in Wolbachia infection intensity following exposure to high temperature (37°C) in both sexes and implied that Wolbachia might survive in room temperature (25°C) better than in high temperature. Experimental results of crowding effects on Wolbachia infection intensity suggested a negative correlation between copepod nauplii and Wolbachia infection intensity. No effect of rearing temperature on the sex ratio was reported although the fecundity was significantly decreased by high temperature. The results showed that Wolbachia infection intensity to be correlated with crowding conditions and was decreased following exposure of elevated temperature. Copyright © 2016 Elsevier Inc. All rights reserved.

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

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

Draft Genome Assembly of a Wolbachia Endosymbiont of Plutella australiana

PubMed Central

Ward, Christopher M.

2017-01-01

ABSTRACT Wolbachia spp. are endosymbiotic bacteria that infect around 50% of arthropods and cause a broad range of effects, including manipulating host reproduction. Here, we present the annotated draft genome assembly of Wolbachia strain wAus, which infects Plutella australiana, a cryptic ally of the major Brassica pest Plutella xylostella (diamondback moth). PMID:29074653

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. © 2012 The Authors. Journal of Evolutionary Biology © 2012 European Society For Evolutionary Biology.

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

Draft Genome Assembly of a Wolbachia Endosymbiont of Plutella australiana.

PubMed

Ward, Christopher M; Baxter, Simon W

2017-10-26

Wolbachia spp. are endosymbiotic bacteria that infect around 50% of arthropods and cause a broad range of effects, including manipulating host reproduction. Here, we present the annotated draft genome assembly of Wolbachia strain wAus, which infects Plutella australiana , a cryptic ally of the major Brassica pest Plutella xylostella (diamondback moth). Copyright © 2017 Ward and Baxter.

Determination of Wolbachia Diversity in Butterflies from Western Ghats, India, by a Multigene Approach

PubMed Central

Salunke, Bipinchandra K.; Salunkhe, Rahul C.; Dhotre, Dhiraj P.; Walujkar, Sandeep A.; Khandagale, Avinash B.; Chaudhari, Rahul; Chandode, Rakesh K.; Ghate, Hemant V.; Patole, Milind S.; Werren, John H.

2012-01-01

Members of the genus Wolbachia are intracellular bacteria that are widespread in arthropods and establish diverse symbiotic associations with their hosts, ranging from mutualism to parasitism. Here we present the first detailed analyses of Wolbachia in butterflies from India with screening of 56 species. Twenty-nine species (52%) representing five families were positive for Wolbachia. This is the first report of Wolbachia infection in 27 of the 29 species; the other two were reported previously. This study also provides the first evidence of infection in the family Papilionidae. A striking diversity was observed among Wolbachia strains in butterfly hosts based on five multilocus sequence typing (MLST) genes, with 15 different sequence types (STs). Thirteen STs are new to the MLST database, whereas ST41 and ST125 were reported earlier. Some of the same host species from this study carried distinctly different Wolbachia strains, whereas the same or different butterfly hosts also harbored closely related Wolbachia strains. Butterfly-associated STs in the Indian sample originated by recombination and point mutation, further supporting the role of both processes in generating Wolbachia diversity. Recombination was detected only among the STs in this study and not in those from the MLST database. Most of the strains were remarkably similar in their wsp genotype, despite divergence in MLST. Only two wsp alleles were found among 25 individuals with complete hypervariable region (HVR) peptide profiles. Although both wsp and MLST show variability, MLST gives better separation between the strains. Completely different STs were characterized for the individuals sharing the same wsp alleles. PMID:22504801

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

PubMed Central

2013-01-01

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

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

Loss of Wolbachia infection during colonisation in the invasive Argentine ant Linepithema humile.

PubMed

Reuter, M; Pedersen, J S; Keller, L

2005-03-01

WOLBACHIA are maternally inherited bacteria, which are very common in arthropods and nematodes. Wolbachia infection may affect host reproduction through feminisation, parthenogenesis, male-killing, cytoplasmic incompatibility and increased fecundity. Previous studies showing discrepancies between the phylogenies of Wolbachia and its arthropod hosts indicate that infection is frequently lost, but the causes of symbiont extinction have so far remained elusive. Here, we report data showing that colonisation of new habitats is a possible mechanism leading to the loss of infection. The presence and prevalence of Wolbachia were studied in three native and eight introduced populations of the Argentine ant Linepithema humile. The screening shows that the symbiont is common in the three native L. humile populations analysed. In contrast, Wolbachia was detected in only one of the introduced populations. The loss of infection associated with colonisation of new habitats may result from drift (founder effect) or altered selection pressures in the new habitat. Furthermore, a molecular phylogeny based on sequences of the Wolbachia wsp gene indicates that L. humile has been infected by a single strain. Horizontal transmission of the symbiont may be important in ants as suggested by the sequence similarity of strains in the three genera Linepithema, Acromyrmex, and Solenopsis native from South and Central America.

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

The effect of virus-blocking Wolbachia on male competitiveness of the dengue vector mosquito, Aedes aegypti.

PubMed

Segoli, Michal; Hoffmann, Ary A; Lloyd, Jane; Omodei, Gavin J; Ritchie, Scott A

2014-12-01

The bacterial endosymbiont Wolbachia blocks the transmission of dengue virus by its vector mosquito Aedes aegypti, and is currently being evaluated for control of dengue outbreaks. Wolbachia induces cytoplasmic incompatibility (CI) that results in the developmental failure of offspring in the cross between Wolbachia-infected males and uninfected females. This increases the relative success of infected females in the population, thereby enhancing the spread of the beneficial bacterium. However, Wolbachia spread via CI will only be feasible if infected males are sufficiently competitive in obtaining a mate under field conditions. We tested the effect of Wolbachia on the competitiveness of A. aegypti males under semi-field conditions. In a series of experiments we exposed uninfected females to Wolbachia-infected and uninfected males simultaneously. We scored the competitiveness of infected males according to the proportion of females producing non-viable eggs due to incompatibility. We found that infected males were equally successful to uninfected males in securing a mate within experimental tents and semi-field cages. This was true for males infected by the benign wMel Wolbachia strain, but also for males infected by the virulent wMelPop (popcorn) strain. By manipulating male size we found that larger males had a higher success than smaller underfed males in the semi-field cages, regardless of their infection status. The results indicate that Wolbachia infection does not reduce the competitiveness of A. aegypti males. Moreover, the body size effect suggests a potential advantage for lab-reared Wolbachia-males during a field release episode, due to their better nutrition and larger size. This may promote Wolbachia spread via CI in wild mosquito populations and underscores its potential use for disease control.

Detection and Phylogenetic Analysis of Wolbachia in the Asiatic Rice Leafroller, Cnaphalocrocis medinalis, in Chinese Populations

PubMed Central

Chai, Huan-Na; Du, Yu-Zhou; Qiu, Bao-Li; Zhai, Bao-Ping

2011-01-01

Wolbachia are a group of intracellular inherited endosymbiontic bacteria infecting a wide range of insects. In this study the infection status of Wolbachia (Rickettsiales: Rickettsiaceae) was measured in the Asiatic rice leafroller, Cnaphalocrocis medinalis (Guenée) (Lepidoptera: Pyralidae), from twenty locations in China by sequencing wsp, ftsZ and 16S rDNA genes. The results showed high infection rates of Wolbachia in C. medinalis populations. Wolbachia was detected in all geographically separate populations; the average infection rate was ∼ 62.5%, and the highest rates were 90% in Wenzhou and Yangzhou populations. The Wolbachia detected in different C. medinalis populations were 100% identical to each other when wsp, ftsZ, and 16S rDNA sequences were compared, with all sequences belonging to the Wolbachia B supergroup. Based on wsp, ftsZ and 16S rDNA sequences of Wolbachia, three phylogenetic trees of similar pattern emerged. This analysis indicated the possibility of inter-species and intra-species horizontal transmission of Wolbachia in different arthropods in related geographical regions. The migration route of C. medinalis in mainland China was also discussed since large differentiation had been found between the wsp sequences of Chinese and Thai populations. PMID:22233324

A computer simulation model of Wolbachia invasion for disease vector population modification.

PubMed

Guevara-Souza, Mauricio; Vallejo, Edgar E

2015-10-05

Wolbachia invasion has been proved to be a promising alternative for controlling vector-borne diseases, particularly Dengue fever. Creating computer models that can provide insight into how vector population modification can be achieved under different conditions would be most valuable for assessing the efficacy of control strategies for this disease. In this paper, we present a computer model that simulates the behavior of native mosquito populations after the introduction of mosquitoes infected with the Wolbachia bacteria. We studied how different factors such as fecundity, fitness cost of infection, migration rates, number of populations, population size, and number of introduced infected mosquitoes affect the spread of the Wolbachia bacteria among native mosquito populations. Two main scenarios of the island model are presented in this paper, with infected mosquitoes introduced into the largest source population and peripheral populations. Overall, the results are promising; Wolbachia infection spreads among native populations and the computer model is capable of reproducing the results obtained by mathematical models and field experiments. Computer models can be very useful for gaining insight into how Wolbachia invasion works and are a promising alternative for complementing experimental and mathematical approaches for vector-borne disease control.

The Effect of Virus-Blocking Wolbachia on Male Competitiveness of the Dengue Vector Mosquito, Aedes aegypti

PubMed Central

Segoli, Michal; Hoffmann, Ary A.; Lloyd, Jane; Omodei, Gavin J.; Ritchie, Scott A.

2014-01-01

Background The bacterial endosymbiont Wolbachia blocks the transmission of dengue virus by its vector mosquito Aedes aegypti, and is currently being evaluated for control of dengue outbreaks. Wolbachia induces cytoplasmic incompatibility (CI) that results in the developmental failure of offspring in the cross between Wolbachia-infected males and uninfected females. This increases the relative success of infected females in the population, thereby enhancing the spread of the beneficial bacterium. However, Wolbachia spread via CI will only be feasible if infected males are sufficiently competitive in obtaining a mate under field conditions. We tested the effect of Wolbachia on the competitiveness of A. aegypti males under semi-field conditions. Methodology/Principal Findings In a series of experiments we exposed uninfected females to Wolbachia-infected and uninfected males simultaneously. We scored the competitiveness of infected males according to the proportion of females producing non-viable eggs due to incompatibility. We found that infected males were equally successful to uninfected males in securing a mate within experimental tents and semi-field cages. This was true for males infected by the benign wMel Wolbachia strain, but also for males infected by the virulent wMelPop (popcorn) strain. By manipulating male size we found that larger males had a higher success than smaller underfed males in the semi-field cages, regardless of their infection status. Conclusions/Significance The results indicate that Wolbachia infection does not reduce the competitiveness of A. aegypti males. Moreover, the body size effect suggests a potential advantage for lab-reared Wolbachia-males during a field release episode, due to their better nutrition and larger size. This may promote Wolbachia spread via CI in wild mosquito populations and underscores its potential use for disease control. PMID:25502564

Sex ratio and Wolbachia infection in the ant Formica exsecta.

PubMed

Keller, L; Liautard, C; Reuter, M; Brown, W D; Sundström, L; Chapuisat, M

2001-08-01

Sex allocation data in social Hymenoptera provide some of the best tests of kin selection, parent-offspring conflict and sex ratio theories. However, these studies critically depend on controlling for confounding ecological factors and on identifying all parties that potentially manipulate colony sex ratio. It has been suggested that maternally inherited parasites may influence sex allocation in social Hymenoptera. If the parasites can influence sex allocation, infected colonies are predicted to invest more resources in females than non-infected colonies, because the parasites are transmitted through females but not males. Prime candidates for such sex ratio manipulation are Wolbachia, because these cytoplasmically transmitted bacteria have been shown to affect the sex ratio of host arthropods by cytoplasmic incompatibility, parthenogenesis, male-killing and feminization. In this study, we tested whether Wolbachia infection is associated with colony sex ratio in two populations of the ant Formica exsecta that have been the subject of extensive sex ratio studies. In these populations colonies specialize in the production of one sex or the other. We found that almost all F. exsecta colonies in both populations are infected with Wolbachia. However, in neither population did we find a significant association in the predicted direction between the prevalence of Wolbachia and colony sex ratio. In particular, colonies with a higher proportion of infected workers did not produce more females. Hence, we conclude that Wolbachia does not seem to alter the sex ratio of its hosts as a means to increase transmission rate in these two populations of ants.

Open release of male mosquitoes infected with a wolbachia biopesticide: field performance and infection containment.

PubMed

O'Connor, Linda; Plichart, Catherine; Sang, Ayo Cheong; Brelsfoard, Corey L; Bossin, Hervé C; Dobson, Stephen L

2012-01-01

Lymphatic filariasis (LF) is a globally significant disease, with 1.3 billion persons in 83 countries at risk. A coordinated effort of administering annual macrofilaricidal prophylactics to the entire at-risk population has succeeded in impacting and eliminating LF transmission in multiple regions. However, some areas in the South Pacific are predicted to persist as transmission sites, due in part to the biology of the mosquito vector, which has led to a call for additional tools to augment drug treatments. Autocidal strategies against mosquitoes are resurging in the effort against invasive mosquitoes and vector borne disease, with examples that include field trials of genetically modified mosquitoes and Wolbachia population replacement. However, critical questions must be addressed in anticipation of full field trials, including assessments of field competitiveness of transfected males and the risk of unintended population replacement. We report the outcome of field experiments testing a strategy that employs Wolbachia as a biopesticide. The strategy is based upon Wolbachia-induced conditional sterility, known as cytoplasmic incompatibility, and the repeated release of incompatible males to suppress a population. A criticism of the Wolbachia biopesticide approach is that unintended female release or horizontal Wolbachia transmission can result in population replacement instead of suppression. We present the outcome of laboratory and field experiments assessing the competitiveness of transfected males and their ability to transmit Wolbachia via horizontal transmission. The results demonstrate that Wolbachia-transfected Aedes polynesiensis males are competitive under field conditions during a thirty-week open release period, as indicated by mark, release, recapture and brood-hatch failure among females at the release site. Experiments demonstrate the males to be 'dead end hosts' for Wolbachia and that methods were adequate to prevent population replacement at the

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

2016-06-08

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. Copyright © 2016 The Author(s). Published by Elsevier Inc. All rights reserved.

Detection of Spiroplasma and Wolbachia in the bacterial gonad community of Chorthippus parallelus.

PubMed

Martínez-Rodríguez, P; Hernández-Pérez, M; Bella, J L

2013-07-01

We have recently detected the endosymbiont Wolbachia in multiple individuals and populations of the grasshopper Chorthippus parallelus (Orthoptera: acrididae). This bacterium induces reproductive anomalies, including cytoplasmic incompatibility. Such incompatibilities may help explain the maintenance of two distinct subspecies of this grasshopper, C. parallelus parallelus and C. parallelus erythropus, which are involved in a Pyrenean hybrid zone that has been extensively studied for the past 20 years, becoming a model system for the study of genetic divergence and speciation. To evaluate whether Wolbachia is the sole bacterial infection that might induce reproductive anomalies, the gonadal bacterial community of individuals from 13 distinct populations of C. parallelus was determined by denaturing gradient gel electrophoresis analysis of bacterial 16S rRNA gene fragments and sequencing. The study revealed low bacterial diversity in the gonads: a persistent bacterial trio consistent with Spiroplasma sp. and the two previously described supergroups of Wolbachia (B and F) dominated the gonad microbiota. A further evaluation of the composition of the gonad bacterial communities was carried out by whole cell hybridization. Our results confirm previous studies of the cytological distribution of Wolbachia in C. parallelus gonads and show a homogeneous infection by Spiroplasma. Spiroplasma and Wolbachia cooccurred in some individuals, but there was no significant association of Spiroplasma with a grasshopper's sex or with Wolbachia infection, although subtle trends might be detected with a larger sample size. This information, together with previous experimental crosses of this grasshopper, suggests that Spiroplasma is unlikely to contribute to sex-specific reproductive anomalies; instead, they implicate Wolbachia as the agent of the observed anomalies in C. parallelus.

The AWED trial (Applying Wolbachia to Eliminate Dengue) to assess the efficacy of Wolbachia-infected mosquito deployments to reduce dengue incidence in Yogyakarta, Indonesia: study protocol for a cluster randomised controlled trial.

PubMed

Anders, Katherine L; Indriani, Citra; Ahmad, Riris Andono; Tantowijoyo, Warsito; Arguni, Eggi; Andari, Bekti; Jewell, Nicholas P; Rances, Edwige; O'Neill, Scott L; Simmons, Cameron P; Utarini, Adi

2018-05-31

Dengue and other arboviruses transmitted by Aedes aegypti mosquitoes, including Zika and chikungunya, present an increasing public health challenge in tropical regions. Current vector control strategies have failed to curb disease transmission, but continue to be employed despite the absence of robust evidence for their effectiveness or optimal implementation. The World Mosquito Program has developed a novel approach to arbovirus control using Ae. aegypti stably transfected with Wolbachia bacterium, with a significantly reduced ability to transmit dengue, Zika and chikungunya in laboratory experiments. Modelling predicts this will translate to local elimination of dengue in most epidemiological settings. This study protocol describes the first trial to measure the efficacy of Wolbachia in reducing dengue virus transmission in the field. The study is a parallel, two-arm, non-blinded cluster randomised controlled trial conducted in a single site in Yogyakarta, Indonesia. The aim is to determine whether large-scale deployment of Wolbachia-infected Ae. aegypti mosquitoes leads to a measurable reduction in dengue incidence in treated versus untreated areas. The primary endpoint is symptomatic, virologically confirmed dengue virus infection of any severity. The 26 km 2 study area was subdivided into 24 contiguous clusters, allocated randomly 1:1 to receive Wolbachia deployments or no intervention. We use a novel epidemiological study design, the cluster-randomised test-negative design trial, in which dengue cases and arbovirus-negative controls are sampled concurrently from among febrile patients presenting to a network of primary care clinics, with case or control status classified retrospectively based on the results of laboratory diagnostic testing. Efficacy is estimated from the odds ratio of Wolbachia exposure distribution (probability of living in a Wolbachia-treated area) among virologically confirmed dengue cases compared to test-negative controls. A secondary

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

2016-07-01

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.

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

Optimal control approach for establishing wMelPop Wolbachia infection among wild Aedes aegypti populations.

PubMed

Campo-Duarte, Doris E; Vasilieva, Olga; Cardona-Salgado, Daiver; Svinin, Mikhail

2018-06-01

Wolbachia-based biocontrol has recently emerged as a potential method for prevention and control of dengue and other vector-borne diseases. Major vector species, such as Aedes aegypti females, when deliberately infected with Wolbachia become less capable of getting viral infections and transmitting the virus to human hosts. In this paper, we propose an explicit sex-structured population model that describes an interaction of uninfected (wild) male and female mosquitoes and those deliberately infected with wMelPop strain of Wolbachia in the same locality. This particular strain of Wolbachia is regarded as the best blocker of dengue and other arboviral infections. However, wMelPop strain of Wolbachia also causes the loss of individual fitness in Aedes aegypti mosquitoes. Our model allows for natural introduction of the decision (or control) variable, and we apply the optimal control approach to simulate wMelPop Wolbachia infestation of wild Aedes aegypti populations. The control action consists in continuous periodic releases of mosquitoes previously infected with wMelPop strain of Wolbachia in laboratory conditions. The ultimate purpose of control is to find a tradeoff between reaching the population replacement in minimum time and with minimum cost of the control effort. This approach also allows us to estimate the number of Wolbachia-carrying mosquitoes to be released in day-by-day control action. The proposed method of biological control is safe to human health, does not contaminate the environment, does not make harm to non-target species, and preserves their interaction with mosquitoes in the ecosystem.

High levels of multiple infections, recombination and horizontal transmission of Wolbachia in the Andricus mukaigawae (Hymenoptera; Cynipidae) communities.

PubMed

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

Wolbachia-mediated parthenogenesis in the predatory thrips Franklinothrips vespiformis (Thysanoptera: Insecta).

PubMed Central

Arakaki, N.; Miyoshi, T.; Noda, H.

2001-01-01

Wolbachia are bacterial endosymbionts in arthropods and filarial nematodes. They cause thelytoky, which is a form of parthenogenesis in which females produce females without males, in hymenopteran insects. Infection of this parthenogenesis-inducing Wolbachia has been restricted to the order Hymenoptera, but was found in another insect order, Thysanoptera. A parthenogenetic colony of a predatory thrips Franklinothrips vespiformis (Aeolothripidae) possessed B-group Wolbachia. Male progeny were produced from this thrips by heat and tetracycline treatments. Males produced motile sperm, which were transferred to the female spermatheca by mating. However, the mating did not affect the sex ratios of the next generation, suggesting that the sperm do not fertilize the eggs. PMID:11375084

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

Hidden Wolbachia diversity in field populations of the European cherry fruit fly, Rhagoletis cerasi (Diptera, Tephritidae).

PubMed

Arthofer, Wolfgang; Riegler, Markus; Schneider, Daniela; Krammer, Martin; Miller, Wolfgang J; Stauffer, Christian

2009-09-01

The European cherry fruit fly Rhagoletis cerasi has been a field model for cytoplasmic incompatibility since the mid 1970s. Two Wolbachia strains were detected in this tephritid species and wCer2 was described as the CI inducing agent dividing European populations into two unidirectional incompatible groups, i.e. southern females produce viable offspring with northern males, whereas the reciprocal cross results in incompatibility. We detected three new Wolbachia strains by sequencing a multitude of plasmids derived from Wolbachia surface protein gene (wsp) polymerase chain reaction (PCR) products. Strain-specific primers were developed allowing individual diagnosis without need for cloning. Hybridization of specific PCR products with a wsp oligonucleotide enhanced the detection limit significantly and revealed the presence of low-titre infections in some strains, in different ontogenetic stages and in adults of different age. We then performed a survey of strain prevalence and infection frequency in eight European regions. wCer1 was fixed in all populations, whereas wCer2 was detected only in the South. wCer3 frequency was the lowest without a clear distribution pattern. The abundance of wCer4 was homogenous across Europe. Like wCer2, wCer5 showed significant differences in spatial distribution. Our new findings of previously undetected and recombinant Wolbachia strains in R. cerasi reveal a major caveat to the research community not to overlook hidden Wolbachia diversity in field populations. Low-titres and geographical variability in Wolbachia diversity are expected to influence the outcome of Wolbachia population dynamics and Wolbachia-based insect population control and may create invasion barriers for expanding and artificially introduced Wolbachia strains.

Parthenogenesis induction by Wolbachia in parasitoids

USDA-ARS?s Scientific Manuscript database

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...

Female Adult Aedes albopictus Suppression by Wolbachia-Infected Male Mosquitoes.

PubMed

Mains, James W; Brelsfoard, Corey L; Rose, Robert I; Dobson, Stephen L

2016-09-23

Dengue, chikungunya and zika viruses are pathogens with an increasing global impact. In the absence of an approved vaccine or therapy, their management relies on controlling the mosquito vectors. But traditional controls are inadequate, and the range of invasive species such as Aedes albopictus (Asian Tiger Mosquito) is expanding. Genetically modified mosquitoes are being tested, but their use has encountered regulatory barriers and public opposition in some countries. Wolbachia bacteria can cause a form of conditional sterility, which can provide an alternative to genetic modification or irradiation. It is unknown however, whether openly released, artificially infected male Ae. albopictus can competitively mate and sterilize females at a level adequate to suppress a field population. Also, the unintended establishment of Wolbachia at the introduction site could result from horizontal transmission or inadvertent female release. In 2014, an Experimental Use Permit from the United States Environmental Protection Agency approved a pilot field trial in Lexington, Kentucky, USA. Here, we present data showing localized reduction of both egg hatch and adult female numbers. The artificial Wolbachia type was not observed to establish in the field. The results are discussed in relation to the applied use of Wolbachia-infected males as a biopesticide to suppress field populations of Ae. albopictus.

Vector competence of Malaysian Aedes albopictus with and without Wolbachia to four dengue virus serotypes.

PubMed

Joanne, Sylvia; Vythilingam, Indra; Teoh, Boon-Teong; Leong, Cherng-Shii; Tan, Kim-Kee; Wong, Meng-Li; Yugavathy, Nava; AbuBakar, Sazaly

2017-09-01

To determine the susceptibility status of Aedes albopictus with and without Wolbachia to the four dengue virus serotypes. Two newly colonised colonies of Ae. albopictus from the wild were used for the study. One colony was naturally infected with Wolbachia while in the other Wolbachia was removed by tetracycline treatment. Both colonies were orally infected with dengue virus-infected fresh blood meal. Dengue virus load was measured using quantitative RT-PCR at four-time intervals in the salivary glands, midguts and ovaries. Wolbachia did not significantly affect Malaysian Ae. albopictus dengue infection or the dissemination rate for all four dengue virus serotypes. Malaysian Ae. albopictus had the highest replication kinetics for DENV-1 and the highest salivary gland and midgut infection rate for DENV-4. Wolbachia, which naturally exists in Malaysian Ae. albopictus, does not significantly affect dengue virus replication. Malaysian Ae. albopictus is susceptible to dengue virus infections and capable of transmitting dengue virus, especially DENV-1 and DENV-4. Removal of Wolbachia from Malaysian Ae. albopictus would not reduce their susceptibility status. © 2017 John Wiley & Sons Ltd.

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

Various Wolbachia genotypes differently influence host Drosophila dopamine metabolism and survival under heat stress conditions.

PubMed

Gruntenko, Nataly Е; Ilinsky, Yury Yu; Adonyeva, Natalya V; Burdina, Elena V; Bykov, Roman A; Menshanov, Petr N; Rauschenbach, Inga Yu

2017-12-28

One of the most widespread prokaryotic symbionts of invertebrates is the intracellular bacteria of Wolbachia genus which can be found in about 50% of insect species. Wolbachia causes both parasitic and mutualistic effects on its host that include manipulating the host reproductive systems in order to increase their transmission through the female germline, and increasing the host fitness. One of the mechanisms, promoting adaptation in biological organisms, is a non-specific neuroendocrine stress reaction. In insects, this reaction includes catecholamines, dopamine, serotonin and octopamine, which act as neurotransmitters, neuromodulators and neurohormones. The level of dopamine metabolism correlates with heat stress resistance in Drosophila adults. To examine Wolbachia effect on Drosophila survival under heat stress and dopamine metabolism we used five strains carrying the nuclear background of interbred Bi90 strain and cytoplasmic backgrounds with different genotype variants of Wolbachia (produced by 20 backcrosses of Bi90 males with appropriate source of Wolbachia). Non-infected Bi90 strain (treated with tetracycline for 3 generations) was used as a control group. We demonstrated that two of five investigated Wolbachia variants promote changes in Drosophila heat stress resistance and activity of enzymes that produce and degrade dopamine, alkaline phosphatase and dopamine-dependent arylalkylamine N-acetyltransferase. What is especially interesting, wMelCS genotype of Wolbachia increases stress resistance and the intensity of dopamine metabolism, whereas wMelPop strain decreases them. wMel, wMel2 and wMel4 genotypes of Wolbachia do not show any effect on the survival under heat stress or dopamine metabolism. L-DOPA treatment, known to increase the dopamine content in Drosophila, levels the difference in survival under heat stress between all studied groups. The genotype of symbiont determines the effect that the symbiont has on the stress resistance of the host

Wolbachia Effects on Rift Valley Virus Infection in Culex tarsalis Mosquitoes

DTIC Science & Technology

2017-04-25

Vector-Borne Disease Section, Division of Communicable Diseases, Center 14" for Infectious Diseases, California Department of Public Health ...Wolbachia. 44" 45" Author Summary 46" An integrated vector management program utilizes several practices, including pesticide 47" application and source...mosquitoes and can block pathogen transmission to humans . 51" Additionally, Wolbachia is maternally-inherited, allowing it to spread quickly through

Longevity-modulating effects of symbiosis: insights from Drosophila-Wolbachia interaction.

PubMed

Maistrenko, Oleksandr M; Serga, Svitlana V; Vaiserman, Alexander M; Kozeretska, Iryna A

2016-11-01

Microbial communities are known to significantly affect various fitness components and survival of their insect hosts, including Drosophila. The composition of symbiotic microbiota has been shown to change with the host's aging. It is unclear whether these changes are caused by the aging process or, vice versa, they affect the host's aging and longevity. Recent findings indicate that fitness and lifespan of Drosophila are affected by endosymbiotic bacteria Wolbachia. These effects, however, are inconsistent and have been reported both to extend and shorten longevity. The main molecular pathways underlying the lifespan-modulating effects of Wolbachia remain unclear, however insulin/insulin-like growth factor, immune deficiency, ecdysteroid synthesis and signaling and c-Jun N-terminal kinase pathways as well as heat shock protein synthesis and autophagy have been proposed to play a role. Here we revise the current evidence that elucidates the impact of Wolbachia endosymbionts on the aging processes in Drosophila.

Effect of naturally occurring Wolbachia in Anopheles gambiae s.l. mosquitoes from Mali on Plasmodium falciparum malaria transmission

PubMed Central

Gomes, Fabio M.; Hixson, Bretta L.; Tyner, Miles D. W.; Ramirez, Jose Luis; Canepa, Gaspar E.; Alves e Silva, Thiago Luiz; Molina-Cruz, Alvaro; Keita, Moussa; Kane, Fouseyni; Traoré, Boïssé; Sogoba, Nafomon; Barillas-Mury, Carolina

2017-01-01

A naturally occurring Wolbachia strain (wAnga-Mali) was identified in mosquitoes of the Anopheles gambiae complex collected in the Malian villages of Dangassa and Kenieroba. Phylogenetic analysis of the nucleotide sequence of two 16S rRNA regions showed that wAnga-Mali clusters with Wolbachia strains from supergroup A and has the highest homology to a Wolbachia strain isolated from cat fleas (Ctenocephalides). wAnga-Mali is different from two Wolbachia strains previously reported in A. gambiae from Burkina Faso (wAnga_VK5_STP and wAnga_VK5_3.1a). Quantitative analysis of Wolbachia and Plasmodium sporozoite infection in field-collected mosquitoes indicates that the prevalence and intensity of Plasmodium falciparum sporozoite infection is significantly lower in Wolbachia-infected females. The presence of Wolbachia in females from a laboratory Anopheles coluzzii (A. gambiae, M form) colony experimentally infected with P. falciparum (NF54 strain) gametocyte cultures slightly enhanced oocyst infection. However, Wolbachia infection significantly reduced the prevalence and intensity of sporozoite infection, as observed in the field. This indicates that wAnga-Mali infection does not limit early stages of Plasmodium infection in the mosquito, but it has a strong deleterious effect on sporozoites and reduces malaria transmission. PMID:29114059

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

PubMed

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

2014-06-01

Wolbachia are widespread in insects and can manipulate host reproduction. Nasonia vitripennis is a widely studied organism with a very high prevalence of Wolbachia infection. To study the effect of Wolbachia infection in Nasonia spp., it is important to obtain noninfected individuals by artificial methods. Current methods that employ sugar water-containing antibiotics can successfully eliminate Wolbachia from the parasitic wasps; however, treatment of at least three generations is required. Here, we describe a novel, feasible, and effective approach to eliminate Wolbachia from N. vitripennis by feeding fly pupae continuously offering antibiotics to Nasonia populations, which shortened the time to eliminate the pathogens to two generations. Additionally, the Wolbachia Uni and CauB strains have obviously different rifampicin-resistance abilities, which is a previously unknown phenomenon. © 2014 Federation of European Microbiological Societies. Published by John Wiley & Sons Ltd. All rights reserved.

Wolbachia significantly impacts the vector competence of Aedes aegypti for Mayaro virus.

PubMed

Pereira, Thiago Nunes; Rocha, Marcele Neves; Sucupira, Pedro Henrique Ferreira; Carvalho, Fabiano Duarte; Moreira, Luciano Andrade

2018-05-02

Wolbachia, an intracellular endosymbiont present in up to 70% of all insect species, has been suggested as a sustainable strategy for the control of arboviruses such as Dengue, Zika and Chikungunya. As Mayaro virus outbreaks have also been reported in Latin American countries, the objective of this study was to evaluate the vector competence of Brazilian field-collected Ae. aegypti and the impact of Wolbachia (wMel strain) upon this virus. Our in vitro studies with Aag2 cells showed that Mayaro virus can rapidly multiply, whereas in wMel-infected Aag2 cells, viral growth was significantly impaired. In addition, C6/36 cells seem to have alterations when infected by Mayaro virus. In vivo experiments showed that field-collected Ae. aegypti mosquitoes are highly permissive to Mayaro virus infection, and high viral prevalence was observed in the saliva. On the other hand, Wolbachia-harboring mosquitoes showed significantly impaired capability to transmit Mayaro virus. Our results suggest that the use of Wolbachia-harboring mosquitoes may represent an effective mechanism for the reduction of Mayaro virus transmission throughout Latin America.

The transcriptome of the mosquito Aedes fluviatilis (Diptera: Culicidae), and transcriptional changes associated with its native Wolbachia infection.

PubMed

Caragata, E P; Pais, F S; Baton, L A; Silva, J B L; Sorgine, M H F; Moreira, L A

2017-01-03

Wolbachia is a bacterial endosymbiont that naturally infects a wide range of insect species, and causes drastic changes to host biology. Stable infections of Wolbachia in mosquitoes can inhibit infection with medically important pathogens such as dengue virus and malaria-causing Plasmodium parasites. However, some native Wolbachia strains can enhance infection with certain pathogens, as is the case for the mosquito Aedes fluviatilis, where infection with Plasmodium gallinaceum is enhanced by the native wFlu Wolbachia strain. To better understand the biological interactions between mosquitoes and native Wolbachia infections, and to investigate the process of pathogen enhancement, we used RNA-Seq to generate the transcriptome of Ae. fluviatilis with and without Wolbachia infection. In total, we generated 22,280,160 Illumina paired-end reads from Wolbachia-infected and uninfected mosquitoes, and used these to make a de novo transcriptome assembly, resulting in 58,013 contigs with a median sequence length of 443 bp and an N50 of 2454 bp. Contigs were annotated through local alignments using BlastX, and associated with both gene ontology and KEGG orthology terms. Through baySeq, we identified 159 contigs that were significantly upregulated due to Wolbachia infection, and 98 that were downregulated. Critically, we saw no changes to Toll or IMD immune gene transcription, but did see evidence that wFlu infection altered the expression of several bacterial recognition genes, and immune-related genes that could influence Plasmodium infection. wFlu infection also had a widespread effect on a number of host physiological processes including protein, carbohydrate and lipid metabolism, and oxidative stress. We then compared our data set with transcriptomic data for other Wolbachia infections in Aedes aegypti, and identified a core set of 15 gene groups associated with Wolbachia infection in mosquitoes. While the scale of transcriptional changes associated with wFlu infection

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

PubMed Central

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

2016-01-01

Abstract The endosymbiotic bacterium Wolbachia pipientis infects many species of insects and has been transinfected into the mosquito Aedes aegypti (L.), the primary vector of dengue virus (DENV). Recently, it has been shown that Wolbachia blocks the replication and transmission of RNA viruses, such as DENV, in a number of mosquito species including Ae. aegypti and Aedes albopictus (Skuse), which is naturally infected with Wolbachia and considered a secondary vector for DENV. The mosquito species Aedes notoscriptus (Skuse) is highly prevalent in Australia, including in areas where DENV outbreaks have been recorded. The mosquito has been implicated in the transmission of Ross River and Barmah Forest viruses, but not DENV. We investigated whether Wolbachia naturally infects this mosquito species and whether it has an impact on the ability of Ae. notoscriptus to transmit DENV. We show, for the first time, that Ae. notoscriptus is naturally infected with a strain of Wolbachia that belongs to supergroup B and is localized only in the ovaries. However, Wolbachia infection in Ae. notoscriptus did not induce resistance to DENV and had no effect on overall DENV infection rate or titer. The presence of a native Wolbachia in Ae. notoscriptus cannot explain why this mosquito is an ineffective vector of DENV. PMID:26721865

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

PubMed

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

2016-03-01

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

Ensuring successful introduction of Wolbachia in natural populations of Aedes aegypti by means of feedback control.

PubMed

Bliman, Pierre-Alexandre; Aronna, M Soledad; Coelho, Flávio C; da Silva, Moacyr A H B

2018-04-01

The control of the spread of dengue fever by introduction of the intracellular parasitic bacterium Wolbachia in populations of the vector Aedes aegypti, is presently one of the most promising tools for eliminating dengue, in the absence of an efficient vaccine. The success of this operation requires locally careful planning to determine the adequate number of individuals carrying the Wolbachia parasite that need to be introduced into the natural population. The introduced mosquitoes are expected to eventually replace the Wolbachia-free population and guarantee permanent protection against the transmission of dengue to human. In this study, we propose and analyze a model describing the fundamental aspects of the competition between mosquitoes carrying Wolbachia and mosquitoes free of the parasite. We then use feedback control techniques to devise an introduction protocol that is proved to guarantee that the population converges to a stable equilibrium where the totality of mosquitoes carry Wolbachia.

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

PubMed Central

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

2013-01-01

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

Identification and prioritization of novel anti-Wolbachia chemotypes from screening a 10,000-compound diversity library

PubMed Central

Johnston, Kelly L.; Cook, Darren A. N.; Berry, Neil G.; David Hong, W.; Clare, Rachel H.; Goddard, Megan; Ford, Louise; Nixon, Gemma L.; O’Neill, Paul M.; Ward, Stephen A.; Taylor, Mark J.

2017-01-01

Lymphatic filariasis and onchocerciasis are two important neglected tropical diseases (NTDs) that cause severe disability. Control efforts are hindered by the lack of a safe macrofilaricidal drug. Targeting the Wolbachia bacterial endosymbionts in these parasites with doxycycline leads to a macrofilaricidal outcome, but protracted treatment regimens and contraindications restrict its widespread implementation. The Anti-Wolbachia consortium aims to develop improved anti-Wolbachia drugs to overcome these barriers. We describe the first screening of a large, diverse compound library against Wolbachia. This whole-organism screen, streamlined to reduce bottlenecks, produced a hit rate of 0.5%. Chemoinformatic analysis of the top 50 hits led to the identification of six structurally diverse chemotypes, the disclosure of which could offer interesting avenues of investigation to other researchers active in this field. An example of hit-to-lead optimization is described to further demonstrate the potential of developing these high-quality hit series as safe, efficacious, and selective anti-Wolbachia macrofilaricides. PMID:28959730

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

PubMed

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

2013-03-01

Tsetse flies (Diptera: Glossinidae) are the sole vectors of African trypanosomes, the causative agent of sleeping sickness in human and nagana in animals. Like most eukaryotic organisms, Glossina species have established symbiotic associations with bacteria. Three main symbiotic bacteria have been found in tsetse flies: Wigglesworthia glossinidia, an obligate symbiotic bacterium, the secondary endosymbiont Sodalis glossinidius and the reproductive symbiont Wolbachia pipientis. In the present review, we discuss recent studies on the detection and characterization of Wolbachia infections in Glossina species, the horizontal transfer of Wolbachia genes to tsetse chromosomes, the ability of this symbiont to induce cytoplasmic incompatibility in Glossina morsitans morsitans and also how new environment-friendly tools for disease control could be developed by harnessing Wolbachia symbiosis. Copyright © 2012 International Atomic Energy Agency. Published by Elsevier Inc. All rights reserved.

Short- and Long-term Evolutionary Dynamics of Bacterial Insertion Sequences: Insights from Wolbachia Endosymbionts

PubMed Central

Cerveau, Nicolas; Leclercq, Sébastien; Leroy, Elodie; Bouchon, Didier; Cordaux, Richard

2011-01-01

Transposable elements (TE) are one of the major driving forces of genome evolution, raising the question of the long-term dynamics underlying their evolutionary success. Long-term TE evolution can readily be reconstructed in eukaryotes, thanks to many degraded copies constituting genomic fossil records of past TE proliferations. By contrast, bacterial genomes usually experience high sequence turnover and short TE retention times, thereby obscuring ancient TE evolutionary patterns. We found that Wolbachia bacterial genomes contain 52–171 insertion sequence (IS) TEs. IS account for 11% of Wolbachia wRi, which is one of the highest IS genomic coverage reported in prokaryotes to date. We show that many IS groups are currently expanding in various Wolbachia genomes and that IS horizontal transfers are frequent among strains, which can explain the apparent synchronicity of these IS proliferations. Remarkably, >70% of Wolbachia IS are nonfunctional. They constitute an unusual bacterial IS genomic fossil record providing direct empirical evidence for a long-term IS evolutionary dynamics following successive periods of intense transpositional activity. Our results show that comprehensive IS annotations have the potential to provide new insights into prokaryote TE evolution and, more generally, prokaryote genome evolution. Indeed, the identification of an important IS genomic fossil record in Wolbachia demonstrates that IS elements are not always of recent origin, contrary to the conventional view of TE evolution in prokaryote genomes. Our results also raise the question whether the abundance of IS fossils is specific to Wolbachia or it may be a general, albeit overlooked, feature of prokaryote genomes. PMID:21940637

Short- and long-term evolutionary dynamics of bacterial insertion sequences: insights from Wolbachia endosymbionts.

PubMed

Cerveau, Nicolas; Leclercq, Sébastien; Leroy, Elodie; Bouchon, Didier; Cordaux, Richard

2011-01-01

Transposable elements (TE) are one of the major driving forces of genome evolution, raising the question of the long-term dynamics underlying their evolutionary success. Long-term TE evolution can readily be reconstructed in eukaryotes, thanks to many degraded copies constituting genomic fossil records of past TE proliferations. By contrast, bacterial genomes usually experience high sequence turnover and short TE retention times, thereby obscuring ancient TE evolutionary patterns. We found that Wolbachia bacterial genomes contain 52-171 insertion sequence (IS) TEs. IS account for 11% of Wolbachia wRi, which is one of the highest IS genomic coverage reported in prokaryotes to date. We show that many IS groups are currently expanding in various Wolbachia genomes and that IS horizontal transfers are frequent among strains, which can explain the apparent synchronicity of these IS proliferations. Remarkably, >70% of Wolbachia IS are nonfunctional. They constitute an unusual bacterial IS genomic fossil record providing direct empirical evidence for a long-term IS evolutionary dynamics following successive periods of intense transpositional activity. Our results show that comprehensive IS annotations have the potential to provide new insights into prokaryote TE evolution and, more generally, prokaryote genome evolution. Indeed, the identification of an important IS genomic fossil record in Wolbachia demonstrates that IS elements are not always of recent origin, contrary to the conventional view of TE evolution in prokaryote genomes. Our results also raise the question whether the abundance of IS fossils is specific to Wolbachia or it may be a general, albeit overlooked, feature of prokaryote genomes.

Harnessing mosquito-Wolbachia symbiosis for vector and disease control.

PubMed

Bourtzis, Kostas; Dobson, Stephen L; Xi, Zhiyong; Rasgon, Jason L; Calvitti, Maurizio; Moreira, Luciano A; Bossin, Hervé C; Moretti, Riccardo; Baton, Luke Anthony; Hughes, Grant L; Mavingui, Patrick; Gilles, Jeremie R L

2014-04-01

Mosquito species, members of the genera Aedes, Anopheles and Culex, are the major vectors of human pathogens including protozoa (Plasmodium sp.), filariae and of a variety of viruses (causing dengue, chikungunya, yellow fever, West Nile). There is lack of efficient methods and tools to treat many of the diseases caused by these major human pathogens, since no efficient vaccines or drugs are available; even in malaria where insecticide use and drug therapies have reduced incidence, 219 million cases still occurred in 2010. Therefore efforts are currently focused on the control of vector populations. Insecticides alone are insufficient to control mosquito populations since reduced susceptibility and even resistance is being observed more and more frequently. There is also increased concern about the toxic effects of insecticides on non-target (even beneficial) insect populations, on humans and the environment. During recent years, the role of symbionts in the biology, ecology and evolution of insect species has been well-documented and has led to suggestions that they could potentially be used as tools to control pests and therefore diseases. Wolbachia is perhaps the most renowned insect symbiont, mainly due to its ability to manipulate insect reproduction and to interfere with major human pathogens thus providing new avenues for pest control. We herein present recent achievements in the field of mosquito-Wolbachia symbiosis with an emphasis on Aedes albopictus. We also discuss how Wolbachia symbiosis can be harnessed for vector control as well as the potential to combine the sterile insect technique and Wolbachia-based approaches for the enhancement of population suppression programs. Copyright © 2013 International Atomic Energy Agency 2013. Published by Elsevier B.V. All rights reserved.

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

PubMed

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

2016-10-01

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

Conflict in the Intracellular Lives of Endosymbionts and Viruses: A Mechanistic Look at Wolbachia-Mediated Pathogen-blocking

PubMed Central

Newton, Irene L. G.; Hardy, Richard W.

2018-01-01

At the forefront of vector control efforts are strategies that leverage host-microbe associations to reduce vectorial capacity. The most promising of these efforts employs Wolbachia, a maternally transmitted endosymbiotic bacterium naturally found in 40% of insects. Wolbachia can spread through a population of insects while simultaneously inhibiting the replication of viruses within its host. Despite successes in using Wolbachia-transfected mosquitoes to limit dengue, Zika, and chikungunya transmission, the mechanisms behind pathogen-blocking have not been fully characterized. Firstly, we discuss how Wolbachia and viruses both require specific host-derived structures, compounds, and processes to initiate and maintain infection. There is significant overlap in these requirements, and infection with either microbe often manifests as cellular stress, which may be a key component of Wolbachia’s anti-viral effect. Secondly, we discuss the current understanding of pathogen-blocking through this lens of cellular stress and develop a comprehensive view of how the lives of Wolbachia and viruses are fundamentally in conflict with each other. A thorough understanding of the genetic and cellular determinants of pathogen-blocking will significantly enhance the ability of vector control programs to deploy and maintain effective Wolbachia-mediated control measures. PMID:29561780

Mission Accomplished? We Need a Guide to the 'Post Release' World of Wolbachia for Aedes-borne Disease Control.

PubMed

Ritchie, Scott A; van den Hurk, Andrew F; Smout, Michael J; Staunton, Kyran M; Hoffmann, Ary A

2018-03-01

Historically, sustained control of Aedes aegypti, the vector of dengue, chikungunya, yellow fever, and Zika viruses, has been largely ineffective. Subsequently, two novel 'rear and release' control strategies utilizing mosquitoes infected with Wolbachia are currently being developed and deployed widely. In the incompatible insect technique, male Aedes mosquitoes, infected with Wolbachia, suppress populations through unproductive mating. In the transinfection strategy, both male and female Wolbachia-infected Ae. aegypti mosquitoes rapidly infect the wild population with Wolbachia, blocking virus transmission. It is critical to monitor the long-term stability of Wolbachia in host populations, and also the ability of this bacterium to continually inhibit virus transmission. Ongoing release and monitoring programs must be future-proofed should political support weaken when these vectors are successfully controlled. Copyright © 2017 Elsevier Ltd. All rights reserved.

Impact of Wolbachia on infection with chikungunya and yellow fever viruses in the mosquito vector Aedes aegypti.

PubMed

van den Hurk, Andrew F; Hall-Mendelin, Sonja; Pyke, Alyssa T; Frentiu, Francesca D; McElroy, Kate; Day, Andrew; Higgs, Stephen; O'Neill, Scott L

2012-01-01

Incidence of disease due to dengue (DENV), chikungunya (CHIKV) and yellow fever (YFV) viruses is increasing in many parts of the world. The viruses are primarily transmitted by Aedes aegypti, a highly domesticated mosquito species that is notoriously difficult to control. When transinfected into Ae. aegypti, the intracellular bacterium Wolbachia has recently been shown to inhibit replication of DENVs, CHIKV, malaria parasites and filarial nematodes, providing a potentially powerful biocontrol strategy for human pathogens. Because the extent of pathogen reduction can be influenced by the strain of bacterium, we examined whether the wMel strain of Wolbachia influenced CHIKV and YFV infection in Ae. aegypti. Following exposure to viremic blood meals, CHIKV infection and dissemination rates were significantly reduced in mosquitoes with the wMel strain of Wolbachia compared to Wolbachia-uninfected controls. However, similar rates of infection and dissemination were observed in wMel infected and non-infected Ae. aegypti when intrathoracic inoculation was used to deliver virus. YFV infection, dissemination and replication were similar in wMel-infected and control mosquitoes following intrathoracic inoculations. In contrast, mosquitoes with the wMelPop strain of Wolbachia showed at least a 10(4) times reduction in YFV RNA copies compared to controls. The extent of reduction in virus infection depended on Wolbachia strain, titer and strain of the virus, and mode of exposure. Although originally proposed for dengue biocontrol, our results indicate a Wolbachia-based strategy also holds considerable promise for YFV and CHIKV suppression.

Impact of Wolbachia on Infection with Chikungunya and Yellow Fever Viruses in the Mosquito Vector Aedes aegypti

PubMed Central

van den Hurk, Andrew F.; Hall-Mendelin, Sonja; Pyke, Alyssa T.; Frentiu, Francesca D.; McElroy, Kate; Day, Andrew; Higgs, Stephen; O'Neill, Scott L.

2012-01-01

Incidence of disease due to dengue (DENV), chikungunya (CHIKV) and yellow fever (YFV) viruses is increasing in many parts of the world. The viruses are primarily transmitted by Aedes aegypti, a highly domesticated mosquito species that is notoriously difficult to control. When transinfected into Ae. aegypti, the intracellular bacterium Wolbachia has recently been shown to inhibit replication of DENVs, CHIKV, malaria parasites and filarial nematodes, providing a potentially powerful biocontrol strategy for human pathogens. Because the extent of pathogen reduction can be influenced by the strain of bacterium, we examined whether the wMel strain of Wolbachia influenced CHIKV and YFV infection in Ae. aegypti. Following exposure to viremic blood meals, CHIKV infection and dissemination rates were significantly reduced in mosquitoes with the wMel strain of Wolbachia compared to Wolbachia-uninfected controls. However, similar rates of infection and dissemination were observed in wMel infected and non-infected Ae. aegypti when intrathoracic inoculation was used to deliver virus. YFV infection, dissemination and replication were similar in wMel-infected and control mosquitoes following intrathoracic inoculations. In contrast, mosquitoes with the wMelPop strain of Wolbachia showed at least a 104 times reduction in YFV RNA copies compared to controls. The extent of reduction in virus infection depended on Wolbachia strain, titer and strain of the virus, and mode of exposure. Although originally proposed for dengue biocontrol, our results indicate a Wolbachia-based strategy also holds considerable promise for YFV and CHIKV suppression. PMID:23133693

Albendazole and antibiotics synergize to deliver short-course anti-Wolbachia curative treatments in preclinical models of filariasis

PubMed Central

Sharma, Raman; Al Jayoussi, Ghaith; Tyrer, Hayley E.; Gamble, Joanne; Hayward, Laura; Priestley, Richard S.; Murphy, Emma A.; Davies, Jill; Waterhouse, David; Cook, Darren A. N.; Clare, Rachel H.; Cassidy, Andrew; Steven, Andrew; Johnston, Kelly L.; McCall, John; Ford, Louise; Hemingway, Janet; Ward, Stephen A.

2017-01-01

Elimination of filariasis requires a macrofilaricide treatment that can be delivered within a 7-day period. Here we have identified a synergy between the anthelmintic albendazole (ABZ) and drugs depleting the filarial endosymbiont Wolbachia, a proven macrofilaricide target, which reduces treatment from several weeks to 7 days in preclinical models. ABZ had negligible effects on Wolbachia but synergized with minocycline or rifampicin (RIF) to deplete symbionts, block embryogenesis, and stop microfilariae production. Greater than 99% Wolbachia depletion following 7-day combination of RIF+ABZ also led to accelerated macrofilaricidal activity. Thus, we provide preclinical proof-of-concept of treatment shortening using antibiotic+ABZ combinations to deliver anti-Wolbachia sterilizing and macrofilaricidal effects. Our data are of immediate public health importance as RIF+ABZ are registered drugs and thus immediately implementable to deliver a 1-wk macrofilaricide. They also suggest that novel, more potent anti-Wolbachia drugs under development may be capable of delivering further treatment shortening, to days rather than weeks, if combined with benzimidazoles. PMID:29078351

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.

Wolbachia Affects Reproduction and Population Dynamics of the Coffee Berry Borer (Hypothenemus hampei): Implications for Biological Control

PubMed Central

Mariño, Yobana A.; Verle Rodrigues, José C.; Bayman, Paul

2017-01-01

Wolbachia are widely distributed endosymbiotic bacteria that influence the reproduction and fitness of their hosts. In recent years the manipulation of Wolbachia infection has been considered as a potential tool for biological control. The coffee berry borer (CBB), Hypothenemus hampei, is the most devastating coffee pest worldwide. Wolbachia infection in the CBB has been reported, but until now the role of Wolbachia in CBB reproduction and fitness has not been tested. To address this issue we reared the CBB in artificial diets with and without tetracycline (0.1% w/v) for ten generations. Tetracycline reduced significantly the relative proportion of Wolbachia in the CBB microbiota from 0.49% to 0.04%. This reduction affected CBB reproduction: females fed with tetracycline had significantly fewer progeny, lower fecundity, and fewer eggs per female. Tetracycline also reduced the population growth rate (λ), net reproductive rate (R0), and mean generation time (T) in CBB; the reduction in population growth was mostly due to variation in fertility, according to life time response experiments (LTREs) analysis. Our results suggest that Wolbachia contribute to the reproductive success of the CBB and their manipulation represents a possible approach to CBB biocontrol mediated by microbiome management. PMID:28085049

Survey of endosymbionts in the Diaphorina citri metagenome and assembly of a Wolbachia wDi draft genome.

PubMed

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.

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

Wolbachia Transfer from Rhagoletis cerasi to Drosophila simulans: Investigating the Outcomes of Host-Symbiont Coevolution

PubMed Central

Riegler, Markus; Charlat, Sylvain; Stauffer, Christian; Merçot, Hervé

2004-01-01

Wolbachia is an endosymbiont of diverse arthropod lineages that can induce various alterations of host reproduction for its own benefice. Cytoplasmic incompatibility (CI) is the most common phenomenon, which results in embryonic lethality when males that bear Wolbachia are mated with females that do not. In the cherry fruit fly, Rhagoletis cerasi, Wolbachia seems to be responsible for previously reported patterns of incompatibility between populations. Here we report on the artificial transfer of two Wolbachia variants (wCer1 and wCer2) from R. cerasi into Drosophila simulans, which was performed with two major goals in mind: first, to isolate wCer1 from wCer2 in order to individually test their respective abilities to induce CI in the new host; and, second, to test the theoretical prediction that recent Wolbachia-host associations should be characterized by high levels of CI, fitness costs to the new host, and inefficient transmission from mothers to offspring. wCer1 was unable to develop in the new host, resulting in its rapid loss after successful injection, while wCer2 was established in the new host. Transmission rates of wCer2 were low, and the infection showed negative fitness effects, consistent with our prediction, but CI levels were unexpectedly lower in the new host. Based on these parameter estimates, neither wCer1 nor wCer2 could be naturally maintained in D. simulans. The experiment thus suggests that natural Wolbachia transfer between species might be restricted by many factors, should the ecological barriers be bypassed. PMID:14711652

Wolbachia transfer from Rhagoletis cerasi to Drosophila simulans: investigating the outcomes of host-symbiont coevolution.

PubMed

Riegler, Markus; Charlat, Sylvain; Stauffer, Christian; Merçot, Hervé

2004-01-01

Wolbachia is an endosymbiont of diverse arthropod lineages that can induce various alterations of host reproduction for its own benefice. Cytoplasmic incompatibility (CI) is the most common phenomenon, which results in embryonic lethality when males that bear Wolbachia are mated with females that do not. In the cherry fruit fly, Rhagoletis cerasi, Wolbachia seems to be responsible for previously reported patterns of incompatibility between populations. Here we report on the artificial transfer of two Wolbachia variants (wCer1 and wCer2) from R. cerasi into Drosophila simulans, which was performed with two major goals in mind: first, to isolate wCer1 from wCer2 in order to individually test their respective abilities to induce CI in the new host; and, second, to test the theoretical prediction that recent Wolbachia-host associations should be characterized by high levels of CI, fitness costs to the new host, and inefficient transmission from mothers to offspring. wCer1 was unable to develop in the new host, resulting in its rapid loss after successful injection, while wCer2 was established in the new host. Transmission rates of wCer2 were low, and the infection showed negative fitness effects, consistent with our prediction, but CI levels were unexpectedly lower in the new host. Based on these parameter estimates, neither wCer1 nor wCer2 could be naturally maintained in D. simulans. The experiment thus suggests that natural Wolbachia transfer between species might be restricted by many factors, should the ecological barriers be bypassed.

Localization and dynamics of Wolbachia infection in Asian citrus psyllid Diaphorina citri, the insect vector of the causal pathogens of Huanglongbing.

PubMed

Ren, Su-Li; Li, Yi-Han; Ou, Da; Guo, Yan-Jun; Qureshi, Jawwad A; Stansly, Philip A; Qiu, Bao-Li

2018-03-23

Wolbachia is a group of intracellular bacteria that infect a wide range of arthropods including the Asian citrus psyllid (ACP), Diaphorina citri Kuwayama. This insect is the vector of Candidatus Liberibacter asiaticus (CLas), the causal pathogen of Huanglongbing or citrus greening disease. Here, we investigated the localization pattern and infection dynamics of Wolbachia in different developmental stages of ACP. Results revealed that all developmental stages of ACP including egg, 1st-5th instar nymphs, and adults of both gender were infected with Wolbachia. FISH visualization of an ACP egg showed that Wolbachia moved from the egg stalk of newly laid eggs to a randomly distributed pattern throughout the egg prior to hatching. The infection rate varied between nymphal instars. The titers of Wolbachia in fourth and fifth instar nymphs were significantly higher than those in the first and second instar nymphs. Wolbachia were scattered in all nymphal stages, but with highest intensity in the U-shaped bacteriome located in the abdomen of the nymph. Wolbachia was confined to two symmetrical organizations in the abdomen of newly emerged female and male adults. The potential mechanisms of Wolbachia infection dynamics are discussed. © 2018 The Authors. MicrobiologyOpen published by John Wiley & Sons Ltd.

Wolbachia supergroups A and B in natural populations of medically important filth flies (diptera: muscidae, calliphoridae, and sarcophagidae) in Thailand.

PubMed

Mingchay, Pichanon; Sai-Ngam, Arkhom; Phumee, Atchara; Bhakdeenuan, Payu; Lorlertthum, Kittitouch; Thavara, Usavadee; Tawatsin, Apiwat; Choochote, Wej; Siriyasatien, Padet

2014-03-01

Filth flies, belonging to suborder Brachycera (Family; Muscidae, Calliphoridae and Sarcophagidae), are a major cause of nuisance and able to transmit pathogens to humans and animals. These insects are distributed worldwide and their populations are increasing especially in sub-tropical and tropical areas. One strategy for controlling insects employs Wolbachia, which is a group of maternally inherited intracellular bacteria, found in many insect species. The bacteria can cause reproductive abnormalities in their hosts, such as cytoplasmic incompatibility, feminization, parthenogenesis, and male lethality. In this study we determined Wolbachia endosymbionts in natural population of medically important flies (42 females and 9 males) from several geographic regions of Thailand. Wolbachia supergroups A or B were detected in 7 of female flies using PCR specific for wsp. Sequence analysis of wsp showed variations between and within the Wolbachia supergroup. Phylogenetics demonstrated that wsp is able to diverge between Wolbachia supergroups A and B. These data should be useful in future Wolbachia-based programs of fly control.

Intragenomic conflict in populations infected by Parthenogenesis Inducing Wolbachia ends with irreversible loss of sexual reproduction

PubMed Central

2010-01-01

Background The maternally inherited, bacterial symbiont, parthenogenesis inducing (PI) Wolbachia, causes females in some haplodiploid insects to produce daughters from both fertilized and unfertilized eggs. The symbionts, with their maternal inheritance, benefit from inducing the production of exclusively daughters, however the optimal sex ratio for the nuclear genome is more male-biased. Here we examine through models how an infection with PI-Wolbachia in a previously uninfected population leads to a genomic conflict between PI-Wolbachia and the nuclear genome. In most natural populations infected with PI-Wolbachia the infection has gone to fixation and sexual reproduction is impossible, specifically because the females have lost their ability to fertilize eggs, even when mated with functional males. Results The PI Wolbachia infection by itself does not interfere with the fertilization process in infected eggs, fertilized infected eggs develop into biparental infected females. Because of the increasingly female-biased sex ratio in the population during a spreading PI-Wolbachia infection, sex allocation alleles in the host that cause the production of more sons are rapidly selected. In haplodiploid species a reduced fertilization rate leads to the production of more sons. Selection for the reduced fertilization rate leads to a spread of these alleles through both the infected and uninfected population, eventually resulting in the population becoming fixed for both the PI-Wolbachia infection and the reduced fertilization rate. Fertilization rate alleles that completely interfere with fertilization ("virginity alleles") will be selected over alleles that still allow for some fertilization. This drives the final resolution of the conflict: the irreversible loss of sexual reproduction and the complete dependence of the host on its symbiont. Conclusions This study shows that dependence among organisms can evolve rapidly due to the resolution of the conflicts between

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.

Wolbachia and dengue virus infection in the mosquito Aedes fluviatilis (Diptera: Culicidae).

PubMed

Silva, Jéssica Barreto Lopes; Magalhães Alves, Debora; Bottino-Rojas, Vanessa; Pereira, Thiago Nunes; Sorgine, Marcos Henrique Ferreira; Caragata, Eric Pearce; Moreira, Luciano Andrade

2017-01-01

Dengue represents a serious threat to human health, with billions of people living at risk of the disease. Wolbachia pipientis is a bacterial endosymbiont common to many insect species. Wolbachia transinfections in mosquito disease vectors have great value for disease control given the bacterium's ability to spread into wild mosquito populations, and to interfere with infections of pathogens, such as dengue virus. Aedes fluviatilis is a mosquito with a widespread distribution in Latin America, but its status as a dengue vector has not been clarified. Ae. fluviatilis is also naturally infected by the wFlu Wolbachia strain, which has been demonstrated to enhance infection with the avian malarial parasite Plasmodium gallinaceum. We performed experimental infections of Ae. fluviatilis with DENV-2 and DENV-3 isolates from Brazil via injection or oral feeding to provide insight into its competence for the virus. We also examined the effect of the native Wolbachia infection on the virus using a mosquito line where the wFlu infection had been cleared by antibiotic treatment. Through RT-qPCR, we observed that Ae. fluviatilis could become infected with both viruses via either method of infection, although at a lower rate than Aedes aegypti, the primary dengue vector. We then detected DENV-2 and DENV-3 in the saliva of injected mosquitoes, and observed that injection of DENV-3-infected saliva produced subsequent infections in naïve Ae. aegypti. However, across our data we observed no difference in prevalence of infection and viral load between Wolbachia-infected and -uninfected mosquitoes, suggesting that there is no effect of wFlu on dengue virus. Our results highlight that Ae. fluviatilis could potentially serve as a dengue vector under the right circumstances, although further testing is required to determine if this occurs in the field.

Wolbachia and dengue virus infection in the mosquito Aedes fluviatilis (Diptera: Culicidae)

PubMed Central

Silva, Jéssica Barreto Lopes; Magalhães Alves, Debora; Bottino-Rojas, Vanessa; Pereira, Thiago Nunes; Sorgine, Marcos Henrique Ferreira; Caragata, Eric Pearce

2017-01-01

Dengue represents a serious threat to human health, with billions of people living at risk of the disease. Wolbachia pipientis is a bacterial endosymbiont common to many insect species. Wolbachia transinfections in mosquito disease vectors have great value for disease control given the bacterium’s ability to spread into wild mosquito populations, and to interfere with infections of pathogens, such as dengue virus. Aedes fluviatilis is a mosquito with a widespread distribution in Latin America, but its status as a dengue vector has not been clarified. Ae. fluviatilis is also naturally infected by the wFlu Wolbachia strain, which has been demonstrated to enhance infection with the avian malarial parasite Plasmodium gallinaceum. We performed experimental infections of Ae. fluviatilis with DENV-2 and DENV-3 isolates from Brazil via injection or oral feeding to provide insight into its competence for the virus. We also examined the effect of the native Wolbachia infection on the virus using a mosquito line where the wFlu infection had been cleared by antibiotic treatment. Through RT-qPCR, we observed that Ae. fluviatilis could become infected with both viruses via either method of infection, although at a lower rate than Aedes aegypti, the primary dengue vector. We then detected DENV-2 and DENV-3 in the saliva of injected mosquitoes, and observed that injection of DENV-3-infected saliva produced subsequent infections in naïve Ae. aegypti. However, across our data we observed no difference in prevalence of infection and viral load between Wolbachia-infected and -uninfected mosquitoes, suggesting that there is no effect of wFlu on dengue virus. Our results highlight that Ae. fluviatilis could potentially serve as a dengue vector under the right circumstances, although further testing is required to determine if this occurs in the field. PMID:28732048

Spiders do not escape reproductive manipulations by Wolbachia

PubMed Central

2011-01-01

Background Maternally inherited bacteria that reside obligatorily or facultatively in arthropods can increase their prevalence in the population by altering their hosts' reproduction. Such reproductive manipulations have been reported from the major arthropod groups such as insects (in particular hymenopterans, butterflies, dipterans and beetles), crustaceans (isopods) and mites. Despite the observation that endosymbiont bacteria are frequently encountered in spiders and that the sex ratio of particular spider species is strongly female biased, a direct relationship between bacterial infection and sex ratio variation has not yet been demonstrated for this arthropod order. Results Females of the dwarf spider Oedothorax gibbosus exhibit considerable variation in the sex ratio of their clutches and were infected with at least three different endosymbiont bacteria capable of altering host reproduction i.e. Wolbachia, Rickettsia and Cardinium. Breeding experiments show that sex ratio variation in this species is primarily maternally inherited and that removal of the bacteria by antibiotics restores an unbiased sex ratio. Moreover, clutches of females infected with Wolbachia were significantly female biased while uninfected females showed an even sex ratio. As female biased clutches were of significantly smaller size compared to non-distorted clutches, killing of male embryos appears to be the most likely manipulative effect. Conclusions This represents to our knowledge the first direct evidence that endosymbiont bacteria, and in particular Wolbachia, might induce sex ratio variation in spiders. These findings are pivotal to further understand the diversity of reproductive phenotypes observed in this arthropod order. PMID:21235755

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

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

USDA-ARS?s Scientific Manuscript database

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...

Suppression of the pelo protein by Wolbachia and its effect on dengue virus in Aedes aegypti

PubMed Central

Asad, Sultan; Hussain, Mazhar; Hugo, Leon; Zhang, Guangmei; Watterson, Daniel

2018-01-01

The endosymbiont Wolbachia is known to block replication of several important arboviruses, including dengue virus (DENV), in the mosquito vector Aedes aegypti. So far, the exact mechanism of this viral inhibition is not fully understood. A recent study in Drosophila melanogaster has demonstrated an interaction between the pelo gene and Drosophila C virus. In this study, we explored the possible involvement of the pelo protein, that is involved in protein translation, in Wolbachia-mediated antiviral response and mosquito-DENV interaction. We found that pelo is upregulated during DENV replication and its silencing leads to reduced DENV virion production suggesting that it facilities DENV replication. However, in the presence of Wolbachia, specifically in female mosquitoes, the pelo protein is downregulated and its subcellular localization is altered, which could contribute to reduction in DENV replication in Ae. aegypti. In addition, we show that the microRNA aae-miR-2940-5p, whose abundance is highly enriched in Wolbachia-infected mosquitoes, might mediate regulation of pelo. Our data reveals identification of pelo as a host factor that is positively involved in DENV replication, and its suppression in the presence of Wolbachia may contribute to virus blocking exhibited by the endosymbiont. PMID:29641562

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

Is Wolbachia participating in the bronchial reactivity of cats with heartworm associated respiratory disease?

PubMed

García-Guasch, L; Caro-Vadillo, A; Manubens-Grau, J; Carretón, E; Morchón, R; Simón, F; Kramer, L H; Montoya-Alonso, J A

2013-09-01

Heartworm associated respiratory disease is a pulmonary syndrome in cats that results from the vascular and parenchymal inflammatory response associated with the arrival and death of Dirofilaria immitis worms into the distal pulmonary arteries. This parasite harbors intracellular Wolbachia, an endosymbiont bacteria. The association between the parasite and the bacteria is obligatory. Some studies suggest the involvement of Wolbachia in the development of the inflammatory reaction and in the polarization of the host immune response against the parasite. Barometric whole-body plethysmography is a non-invasive pulmonary function test that allows a dynamic study of breathing patterns and is useful to study airway disease and the response to different treatments. The aim of this prospective non-blinded study was to compare the influence of Wolbachia upon the respiratory function variables in a population of cats seropositive to D. immitis by use of Barometric whole-body plethysmography. Fourteen seropositive cats to Wolbachia and eight seronegative cats were put into the plethysmograph chamber and different respiratory variables were measured. The results were analyzed and compared between the two groups of animals. Significant differences were found for bronchoconstriction index variables PAU (pause) (P-value<0.05) and Penh (enhanced pause) (P-value<0.05). The results obtained in our study suggest that Wolbachia seems to produce a greater acute inflammatory response at bronchial, vascular and parenchymal level worsening the state of broncho-reactivity associated with the presence of seropositivity to D. immitis in cats. Copyright © 2013 Elsevier B.V. All rights reserved.

Molecular detection and identification of Wolbachia in three species of the genus Lutzomyia on the Colombian Caribbean coast.

PubMed

Vivero, Rafael José; Cadavid-Restrepo, Gloria; Herrera, Claudia Ximena Moreno; Soto, Sandra I Uribe

2017-02-28

The hematophagous habits of insects belonging to the genus Lutzomyia (Diptera: Psychodidae), as well as their role as biological vectors of Leishmania species, make their presence an indication of infection risk. In the present study, seven species of Lutzomyia were identified and screened for natural infections with Wolbachia. Collection of sand flies was done in an endemic focus of leishmaniasis on the Colombian Caribbean coast (Department of Sucre, Ovejas municipality). DNA collected from Lutzomyia species was evaluated with PCR for wsp gene amplification to screen for bacterial infection. Endosymbiotic Wolbachia was found in three species: Lutzomyia c. cayennensis, Lutzomyia dubitans and Lutzomyia evansi. Two Wolbachia strains (genotypes) were found in Lutzomyia spp. These genotypes were previously unknown in dipteran insects. The wLev strain was found in Lutzomyia dubitans, L. c. cayennensis and L. evansi and the wLcy strain was found only in L. c. cayennensis. Genetic analysis indicated that the Wolbachia strains wLcy and wLev belong to the B Supergroup. This study provides evidence of infections of more than one strain of Wolbachia in L. c. cayennensis.

How does competition among wild type mosquitoes influence the performance of Aedes aegypti and dissemination of Wolbachia pipientis?

PubMed Central

de Oliveira, Suellen; Villela, Daniel Antunes Maciel; Dias, Fernando Braga Stehling; Moreira, Luciano Andrade

2017-01-01

Background Wolbachia has been deployed in several countries to reduce transmission of dengue, Zika and chikungunya viruses. During releases, Wolbachia-infected females are likely to lay their eggs in local available breeding sites, which might already be colonized by local Aedes sp. mosquitoes. Therefore, there is an urgent need to estimate the deleterious effects of intra and interspecific larval competition on mosquito life history traits, especially on the duration of larval development time, larval mortality and adult size. Methodology/principal findings Three different mosquito populations were used: Ae. aegypti infected with Wolbachia (wMelBr strain), wild Ae. aegypti and wild Ae. albopictus. A total of 21 treatments explored intra and interspecific larval competition with varying larval densities, species proportions and food levels. Each treatment had eight replicates with two distinct food levels: 0.25 or 0.50 g of Chitosan and fallen avocado leaves. Overall, overcrowding reduced fitness correlates of the three populations. Ae. albopictus larvae presented lower larval mortality, shorter development time to adult and smaller wing sizes than Ae. aegypti. The presence of Wolbachia had a slight positive effect on larval biology, since infected individuals had higher survivorship than uninfected Ae. aegypti larvae. Conclusions/significance In all treatments, Ae. albopictus outperformed both wild Ae. aegypti and the Wolbachia-infected group in larval competition, irrespective of larval density and the amount of food resources. The major force that can slow down Wolbachia invasion is the population density of wild mosquitoes. Given that Ae. aegypti currently dominates in Rio, in comparison with Ae. albopictus frequency, additional attention must be given to the population density of Ae. aegypti during releases to increase the likelihood of Wolbachia invasion. PMID:28991902

Immunohistological studies on neoplasms of female and male Onchocerca volvulus: filarial origin and absence of Wolbachia from tumor cells

PubMed Central

BRATTIG, N. W.; HOERAUF, A.; FISCHER, P. U.; LIEBAU, E.; BANDI, C.; DEBRAH, A.; BÜTTNER, M.; BÜTTNER, D. W.

2010-01-01

SUMMARY Up to 5% of untreated female Onchocerca volvulus filariae develop potentially fatal pleomorphic neoplasms, whose incidence is increased following ivermectin treatment. We studied the occurrence of 8 filarial proteins and of Wolbachia endobacteria in the tumor cells. Onchocercomas from patients, untreated and treated with antibiotics and anthelminthics, were examined by immunohistology. Neoplasms were diagnosed in 112 of 3587 female and in 2 of 1570 male O. volvulus. The following proteins and other compounds of O. volvulus were expressed in the cells of the neoplasms: glutathione S-transferase 1, lysosomal aspartic protease, cAMP-dependent protein kinase, alpha-enolase, aspartate aminotransferase, ankyrin E1, tropomyosin, heat shock protein 60, transforming growth factor-beta, and prostaglandin E2. These findings prove the filarial origin of the neoplasms and confirm the pleomorphism of the tumor cells. Signs indicating malignancy of the neoplasms are described. Wolbachia were observed in the hypodermis, oocytes, and embryos of tumor-harbouring filariae using antibodies against Wolbachia surface protein, Wolbachia HtrA-type serine protease, and Wolbachia aspartate aminotransferase. In contrast, Wolbachia were not found in the cells of the neoplasms. Further, neoplasm-containing worms were not observed after more than 10 months after the start of sufficient treatment with doxycycline or doxycycline plus ivermectin. PMID:20199697

Infection Density Dynamics and Phylogeny of Wolbachia Associated with Coconut Hispine Beetle, Brontispa longissima (Gestro) (Coleoptera: Chrysomelidae), by Multilocus Sequence Type (MLST) Genotyping.

PubMed

Ali, Habib; Muhammad, Abrar; Hou, Youming

2018-05-28

The intracellular bacterium Wolbachia pipientis is widespread in arthropods. Recently, possibilities of novel Wolbachia -mediated hosts, their distribution, and natural rate have been anticipated, and the coconut leaf beetle Brontispa longissima (Gestro) (Coleoptera: Chrysomelidae), which has garnered attention as a serious pest of palms, was subjected to this interrogation. By adopting Wolbachia surface protein ( wsp ) and multilocus sequence type (MLST) genotypic systems, we determined the Wolbachia infection density within host developmental stages, body parts, and tissues, and the results revealed that all the tested samples of B. longissima were infected with the same Wolbachia strain (wLog), suggesting complete vertical transmission. The MLST profile elucidated two new alleles ( ftsZ -234 and coxA-266) that define a new sequence type (ST-483), which indicates the particular genotypic association of B. longissima and Wolbachia . The quantitative real-time PCR analysis revealed a higher infection density in the eggs and adult stage, followed by the abdomen and reproductive tissues, respectively. However, no significant differences were observed in the infection density between sexes. Moreover, the wsp and concatenated MLST alignment analysis of this study with other known Wolbachia-mediated arthropods revealed similar clustering with distinct monophyletic supergroup B. This is the first comprehensive report on the prevalence, infection dynamics, and phylogeny of the Wolbachia endosymbiont in B. longissima , which demonstrated that Wolbachia is ubiquitous across all developmental stages and distributed in the entire body of B. longissima . Understanding the Wolbachia infection dynamics would provide useful insight to build a framework for future investigations, understand its impacts on host physiology, and exploit it as a potential biocontrol agent.

Effects of Larval Nutrition on Wolbachia-Based Dengue Virus Interference in Aedes aegypti (Diptera: Culicidae).

PubMed

Kho, Elise A; Hugo, Leon E; Lu, Guangjin; Smith, David D; Kay, Brian H

2016-07-01

In order to assess the broad-scale applicability of field releases of Wolbachia for the biological control of insect-transmitted diseases, we determined the relationship between the larval diet of Aedes aegypti L. mosquitoes infected with Wolbachia strains and their susceptibility to dengue virus (DENV) infection via intrathoracic injection and oral inoculation. Larvae were reared on diets that varied in the quantity of food which had the effect of modifying development time and adult body size. Wolbachia wMel infection was associated with highly significant reductions in dengue serotype 2 (DENV-2) infection rates of between 80 and 97.5% following intrathoracic injection of adults emerging from three diet levels. Reductions were 100% in two diet level treatments following oral inoculation. Similarly, wMelPop infection was associated with highly significant reductions in DENV-2 infection rates of between 95 and 100% for intrathoracic injection and 97.5 and 100% for oral inoculation across diet level treatments. Larval diet level had no significant effect on DENV-2 infection rates in the presence of Wolbachia infection in mosquitoes that were intrathoracically injected with the virus. This indicates that the effectiveness of Wolbachia on vector competence disruption within Ae. aegypti is unlikely to be compromised by variable larval nutrition in field settings. © The Authors 2016. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

Diversity and distribution of Wolbachia in relation to geography, host plant affiliation and life cycle of a heterogonic gall wasp.

PubMed

Schuler, Hannes; Egan, Scott P; Hood, Glen R; Busbee, Robert W; Driscoe, Amanda L; Ott, James R

2018-03-27

The maternally inherited endosymbiont Wolbachia is widespread in arthropods and nematodes and can play an important role in the ecology and evolution of its host through reproductive manipulation. Here, we survey Wolbachia in Belonocnema treatae, a widely distributed North American cynipid gall forming wasp that exhibits regional host specialization on three species of oaks and alternation of sexually and asexually reproducing generations. We investigated whether patterns of Wolbachia infection and diversity in B. treatae are associated with the insect's geographic distribution, host plant association, life cycle, and mitochondrial evolutionary history. Screening of 463 individuals from 23 populations including sexual and asexual generations from all three host plants across the southern U.S. showed an average infection rate of 56% with three common Wolbachia strains: wTre1-3 and an additional rare variant wTre4. Phylogenetic analysis based on wsp showed that these strains are unrelated and likely independently inherited. We found no difference in Wolbachia infection frequency among host plant associated populations or between the asexual and sexual generations, or between males and females of the sexual generation. Partially incomplete Wolbachia transmission rates might explain the occurrence of uninfected individuals. A parallel analysis of the mitochondrial cytochrome oxidase I gene in B. treatae showed high mtDNA haplotype diversity in both infected and uninfected populations suggesting an ancestral infection by Wolbachia as well as a clear split between eastern and western B. treatae mtDNA clades with a sequence divergence of > 6%. The strain wTre1 was present almost exclusively in the western clade while wTre2 and wTre3 occur almost exclusively in eastern populations. In contrast, the same strains co-occur as double-infections in Georgia and triple-infections in two populations in central Florida. The diversity of Wolbachia across geographically and

Effects of co-occurring Wolbachia and Spiroplasma endosymbionts on the Drosophila immune response against insect pathogenic and non-pathogenic bacteria.

PubMed

Shokal, Upasana; Yadav, Shruti; Atri, Jaishri; Accetta, Julia; Kenney, Eric; Banks, Katherine; Katakam, Akash; Jaenike, John; Eleftherianos, Ioannis

2016-02-09

Symbiotic interactions between microbes and animals are common in nature. Symbiotic organisms are particularly common in insects and, in some cases, they may protect their hosts from pathogenic infections. Wolbachia and Spiroplasma endosymbionts naturally inhabit various insects including Drosophila melanogaster fruit flies. Therefore, this symbiotic association is considered an excellent model to investigate whether endosymbiotic bacteria participate in host immune processes against certain pathogens. Here we have investigated whether the presence of Wolbachia alone or together with Spiroplasma endosymbionts in D. melanogaster adult flies affects the immune response against the virulent insect pathogen Photorhabdus luminescens and against non-pathogenic Escherichia coli bacteria. We found that D. melanogaster flies carrying no endosymbionts, those carrying both Wolbachia and Spiroplasma, and those containing Wolbachia only had similar survival rates after infection with P. luminescens or Escherichia coli bacteria. However, flies carrying both endosymbionts or Wolbachia only contained higher numbers of E. coli cells at early time-points post infection than flies without endosymbiotic bacteria. Interestingly, flies containing Wolbachia only had lower titers of this endosymbiont upon infection with the pathogen P. luminescens than uninfected flies of the same strain. We further found that the presence of Wolbachia and Spiroplasma in D. melanogaster up-regulated certain immune-related genes upon infection with P. luminescens or E. coli bacteria, but it failed to alter the phagocytic ability of the flies toward E. coli inactive bioparticles. Our results suggest that the presence of Wolbachia and Spiroplasma in D. melanogaster can modulate immune signaling against infection by certain insect pathogenic and non-pathogenic bacteria. Results from such studies are important for understanding the molecular basis of the interactions between endosymbiotic bacteria of insects

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. © 2015 The Author(s) Published by the Royal Society. All rights reserved.

Wolbachia in Parasitoids Attacking Native European and Introduced Eastern Cherry Fruit Flies in Europe.

PubMed

Schuler, Hannes; Kern, Peter; Arthofer, Wolfgang; Vogt, Heidrun; Fischer, Maximilian; Stauffer, Christian; Riegler, Markus

2016-12-01

The eastern cherry fruit fly, Rhagoletis cingulata Loew (Diptera: Tephritidae), is an economically important pest of cherries in North America. In 1983 it was first reported in Europe where it shares its ecological niche with the native European cherry fruit fly, Rhagoletis cerasi L. (Diptera: Tephritidae). Their coexistence in Europe led to the recent horizontal transmission of the Wolbachia strain wCer1 from R. cerasi to R. cingulata Horizontal Wolbachia transmission is mediated by either sharing of ecological niches or by interacting species such as parasitoids. Here we describe for the first time that two braconid wasps, Psyttalia rhagoleticola Sachtleben (Hymenoptera: Braconidae) and Utetes magnus Fischer (Hymenoptera: Braconidae), naturally parasitizing R. cerasi, use the invasive R. cingulata in Europe as a new host. In contrast, no parasitoids that parasitize R. cingulata in its native American range were detected in the introduced European range. Diagnostic Wolbachia PCR screening and sequence analyses demonstrated that all P. rhagoleticola individuals were infected with the newly described Wolbachia strain wRha while all U. magnus individuals were uninfected. wRha is different from wCer1 but had an Wolbachia surface protein (wsp) gene sequence that was identical to wCer2 of R. cerasi and wCin2 of R. cingulata. However, multi locus sequence typing revealed differences in all loci between wRha and the tephritid's strains. The horizontal transmission of wCer1 between the two tephritid species did not result in fixed heritable infections in the parasitoids. However, the parasitoids may have acted as a transient wCer1 vector. © The Authors 2016. Published by Oxford University Press on behalf of Entomological Society of America.

Co-evolution between an Endosymbiont and Its Nematode Host: Wolbachia Asymmetric Posterior Localization and AP Polarity Establishment

PubMed Central

Landmann, Frederic; Foster, Jeremy M.; Michalski, Michelle L.; Slatko, Barton E.; Sullivan, William

2014-01-01

While bacterial symbionts influence a variety of host cellular responses throughout development, there are no documented instances in which symbionts influence early embryogenesis. Here we demonstrate that Wolbachia, an obligate endosymbiont of the parasitic filarial nematodes, is required for proper anterior-posterior polarity establishment in the filarial nematode B. malayi. Characterization of pre- and post-fertilization events in B. malayi reveals that, unlike C. elegans, the centrosomes are maternally derived and produce a cortical-based microtubule organizing center prior to fertilization. We establish that Wolbachia rely on these cortical microtubules and dynein to concentrate at the posterior cortex. Wolbachia also rely on PAR-1 and PAR-3 polarity cues for normal concentration at the posterior cortex. Finally, we demonstrate that Wolbachia depletion results in distinct anterior-posterior polarity defects. These results provide a striking example of endosymbiont-host co-evolution operating on the core initial developmental event of axis determination. PMID:25165813

Antibiotic treatment leads to the elimination of Wolbachia endosymbionts and sterility in the diplodiploid collembolan Folsomia candida

PubMed Central

Pike, Nathan; Kingcombe, Rachel

2009-01-01

Background Wolbachia is an extremely widespread bacterial endosymbiont of arthropods and nematodes that causes a variety of reproductive peculiarities. Parthenogenesis is one such peculiarity but it has been hypothesised that this phenomenon may be functionally restricted to organisms that employ haplodiploid sex determination. Using two antibiotics, tetracycline and rifampicin, we attempted to eliminate Wolbachia from the diplodiploid host Folsomia candida, a species of springtail which is a widely used study organism. Results Molecular assays confirmed that elimination of Wolbachia was successfully achieved through continuous exposure of populations (over two generations and several weeks) to rifampicin administered as 2.7% dry weight of their yeast food source. The consequence of this elimination was total sterility of all individuals, despite the continuation of normal egg production. Conclusion Microbial endosymbionts play an obligatory role in the reproduction of their diplodiploid host, most likely one in which the parthenogenetic process is facilitated by Wolbachia. A hitherto unknown level of host-parasite interdependence is thus recorded. PMID:19698188

Fine-scale landscape genomics helps explain the slow spatial spread of Wolbachia through the Aedes aegypti population in Cairns, Australia.

PubMed

Schmidt, Thomas L; Filipović, Igor; Hoffmann, Ary A; Rašić, Gordana

2018-05-01

The endosymbiotic bacterium Wolbachia suppresses the capacity for arbovirus transmission in the mosquito Aedes aegypti, and can spread spatially through wild mosquito populations following local introductions. Recent introductions in Cairns, Australia have demonstrated slower than expected spatial spread. Potential reasons for this include: (i) barriers to Ae. aegypti dispersal; (ii) higher incidence of long-range dispersal; and (iii) intergenerational loss of Wolbachia. We investigated these three potential factors using genome-wide single-nucleotide polymorphisms (SNPs) and an assay for the Wolbachia infection wMel in 161 Ae. aegypti collected from Cairns in 2015. We detected a small but significant barrier effect of Cairns highways on Ae. aegypti dispersal using distance-based redundancy analysis and patch-based simulation analysis. We detected a pair of putative full-siblings in ovitraps 1312 m apart, indicating long-distance female movement likely mediated by human transport. We also found a pair of full-siblings of different infection status, indicating intergenerational loss of Wolbachia in the field. These three factors are all expected to contribute to the slow spread of Wolbachia through Ae. aegypti populations, though from our results it is unclear whether Wolbachia loss and long-distance movement are sufficiently common to reduce the speed of spatial spread appreciably. Our findings inform the strategic deployment of Wolbachia-infected mosquitoes during releases, and show how parameter estimates from laboratory studies may differ from those estimated using field data. Our landscape genomics approach can be extended to other host/symbiont systems that are being considered for biocontrol.

Local introduction and heterogeneous spatial spread of dengue-suppressing Wolbachia through an urban population of Aedes aegypti

PubMed Central

Schmidt, Tom L.; Barton, Nicholas H.; Rašić, Gordana; Turley, Andrew P.; Montgomery, Brian L.; Iturbe-Ormaetxe, Inaki; Cook, Peter E.; Ryan, Peter A.; Ritchie, Scott A.; Hoffmann, Ary A.; O’Neill, Scott L.

2017-01-01

Dengue-suppressing Wolbachia strains are promising tools for arbovirus control, particularly as they have the potential to self-spread following local introductions. To test this, we followed the frequency of the transinfected Wolbachia strain wMel through Ae. aegypti in Cairns, Australia, following releases at 3 nonisolated locations within the city in early 2013. Spatial spread was analysed graphically using interpolation and by fitting a statistical model describing the position and width of the wave. For the larger 2 of the 3 releases (covering 0.97 km2 and 0.52 km2), we observed slow but steady spatial spread, at about 100–200 m per year, roughly consistent with theoretical predictions. In contrast, the smallest release (0.11 km2) produced erratic temporal and spatial dynamics, with little evidence of spread after 2 years. This is consistent with the prediction concerning fitness-decreasing Wolbachia transinfections that a minimum release area is needed to achieve stable local establishment and spread in continuous habitats. Our graphical and likelihood analyses produced broadly consistent estimates of wave speed and wave width. Spread at all sites was spatially heterogeneous, suggesting that environmental heterogeneity will affect large-scale Wolbachia transformations of urban mosquito populations. The persistence and spread of Wolbachia in release areas meeting minimum area requirements indicates the promise of successful large-scale population transformation. PMID:28557993

Local introduction and heterogeneous spatial spread of dengue-suppressing Wolbachia through an urban population of Aedes aegypti.

PubMed

Schmidt, Tom L; Barton, Nicholas H; Rašić, Gordana; Turley, Andrew P; Montgomery, Brian L; Iturbe-Ormaetxe, Inaki; Cook, Peter E; Ryan, Peter A; Ritchie, Scott A; Hoffmann, Ary A; O'Neill, Scott L; Turelli, Michael

2017-05-01

Dengue-suppressing Wolbachia strains are promising tools for arbovirus control, particularly as they have the potential to self-spread following local introductions. To test this, we followed the frequency of the transinfected Wolbachia strain wMel through Ae. aegypti in Cairns, Australia, following releases at 3 nonisolated locations within the city in early 2013. Spatial spread was analysed graphically using interpolation and by fitting a statistical model describing the position and width of the wave. For the larger 2 of the 3 releases (covering 0.97 km2 and 0.52 km2), we observed slow but steady spatial spread, at about 100-200 m per year, roughly consistent with theoretical predictions. In contrast, the smallest release (0.11 km2) produced erratic temporal and spatial dynamics, with little evidence of spread after 2 years. This is consistent with the prediction concerning fitness-decreasing Wolbachia transinfections that a minimum release area is needed to achieve stable local establishment and spread in continuous habitats. Our graphical and likelihood analyses produced broadly consistent estimates of wave speed and wave width. Spread at all sites was spatially heterogeneous, suggesting that environmental heterogeneity will affect large-scale Wolbachia transformations of urban mosquito populations. The persistence and spread of Wolbachia in release areas meeting minimum area requirements indicates the promise of successful large-scale population transformation.

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

USDA-ARS?s Scientific Manuscript database

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...

Wolbachia infections and superinfections in cytoplasmically incompatible populations of the European cherry fruit fly Rhagoletis cerasi (Diptera, Tephritidae).

PubMed

Riegler, Markus; Stauffer, Christian

2002-11-01

Wolbachia is an obligately intracellular, maternally inherited bacterium which has been detected in many arthropods. Wolbachia infections disperse in host populations by mechanisms such as cytoplasmic incompatibility (CI). CI leads to embryonic mortality which occurs when infected males mate with uninfected females or females with a different Wolbachia strain. Populations of the European cherry fruit fly Rhagoletis cerasi (Diptera, Tephritidae) were found to be infected by two different Wolbachia strains, wCer1 and wCer2. Superinfections with both strains occurred throughout southern and central Europe and infections with wCer1 were found in northern, western and eastern Europe. Strong unidirectional CI between European populations of R. cerasi were first reported in the 1970s. From the conformity in the recent geographical distribution of the Wolbachia infections and the CI expression patterns found 25 years ago it was deduced that wCer2 potentially causes CI in R. cerasi. The comparison of the geographical distributions indicated that wCer1 + 2 must have spread into wCer1-infected populations in some areas. In other regions, a spread of wCer1 + 2 was probably prevented by dispersal barriers. There, a sharp transition from infected to superinfected populations suggested regional isolation between wCer1 and wCer1 + 2-infected populations.

Serendipitous discovery of Wolbachia genomes in multiple Drosophila species.

PubMed

Salzberg, Steven L; Dunning Hotopp, Julie C; Delcher, Arthur L; Pop, Mihai; Smith, Douglas R; Eisen, Michael B; Nelson, William C

2005-01-01

The Trace Archive is a repository for the raw, unanalyzed data generated by large-scale genome sequencing projects. The existence of this data offers scientists the possibility of discovering additional genomic sequences beyond those originally sequenced. In particular, if the source DNA for a sequencing project came from a species that was colonized by another organism, then the project may yield substantial amounts of genomic DNA, including near-complete genomes, from the symbiotic or parasitic organism. By searching the publicly available repository of DNA sequencing trace data, we discovered three new species of the bacterial endosymbiont Wolbachia pipientis in three different species of fruit fly: Drosophila ananassae, D. simulans, and D. mojavensis. We extracted all sequences with partial matches to a previously sequenced Wolbachia strain and assembled those sequences using customized software. For one of the three new species, the data recovered were sufficient to produce an assembly that covers more than 95% of the genome; for a second species the data produce the equivalent of a 'light shotgun' sampling of the genome, covering an estimated 75-80% of the genome; and for the third species the data cover approximately 6-7% of the genome. The results of this study reveal an unexpected benefit of depositing raw data in a central genome sequence repository: new species can be discovered within this data. The differences between these three new Wolbachia genomes and the previously sequenced strain revealed numerous rearrangements and insertions within each lineage and hundreds of novel genes. The three new genomes, with annotation, have been deposited in GenBank.

Artificial selection on ant female caste ratio uncovers a link between female-biased sex ratios and infection by Wolbachia endosymbionts.

PubMed

Pontieri, L; Schmidt, A M; Singh, R; Pedersen, J S; Linksvayer, T A

2017-02-01

Social insect sex and caste ratios are well-studied targets of evolutionary conflicts, but the heritable factors affecting these traits remain unknown. To elucidate these factors, we carried out a short-term artificial selection study on female caste ratio in the ant Monomorium pharaonis. Across three generations of bidirectional selection, we observed no response for caste ratio, but sex ratios rapidly became more female-biased in the two replicate high selection lines and less female-biased in the two replicate low selection lines. We hypothesized that this rapid divergence for sex ratio was caused by changes in the frequency of infection by the heritable bacterial endosymbiont Wolbachia, because the initial breeding stock varied for Wolbachia infection, and Wolbachia is known to cause female-biased sex ratios in other insects. Consistent with this hypothesis, the proportions of Wolbachia-infected colonies in the selection lines changed rapidly, mirroring the sex ratio changes. Moreover, the estimated effect of Wolbachia on sex ratio (~13% female bias) was similar in colonies before and during artificial selection, indicating that this Wolbachia effect is likely independent of the effects of artificial selection on other heritable factors. Our study provides evidence for the first case of endosymbiont sex ratio manipulation in a social insect. © 2016 European Society For Evolutionary Biology. Journal of Evolutionary Biology © 2016 European Society For Evolutionary Biology.

Wolbachia Protein TomO Targets nanos mRNA and Restores Germ Stem Cells in Drosophila Sex-lethal Mutants.

PubMed

Ote, Manabu; Ueyama, Morio; Yamamoto, Daisuke

2016-09-12

Wolbachia, endosymbiotic bacteria prevalent in invertebrates, manipulate their hosts in a variety of ways: they induce cytoplasmic incompatibility, male lethality, male-to-female transformation, and parthenogenesis. However, little is known about the molecular basis for host manipulation by these bacteria. In Drosophila melanogaster, Wolbachia infection makes otherwise sterile Sex-lethal (Sxl) mutant females capable of producing mature eggs. Through a functional genomic screen for Wolbachia genes with growth-inhibitory effects when expressed in cultured Drosophila cells, we identified the gene WD1278 encoding a novel protein we call toxic manipulator of oogenesis (TomO), which phenocopies some of the Wolbachia effects in Sxl mutant D. melanogaster females. We demonstrate that TomO enhances the maintenance of germ stem cells (GSCs) by elevating Nanos (Nos) expression via its interaction with nos mRNA, ultimately leading to the restoration of germ cell production in Sxl mutant females that are otherwise without GSCs. Copyright © 2016 Elsevier Ltd. All rights reserved.

Evolutionary Significance of Wolbachia-to-Animal Horizontal Gene Transfer: Female Sex Determination and the f Element in the Isopod Armadillidium vulgare.

PubMed

Cordaux, Richard; Gilbert, Clément

2017-07-21

An increasing number of horizontal gene transfer (HGT) events from bacteria to animals have been reported in the past years, many of which involve Wolbachia bacterial endosymbionts and their invertebrate hosts. Most transferred Wolbachia genes are neutrally-evolving fossils embedded in host genomes. A remarkable case of Wolbachia HGT for which a clear evolutionary significance has been demonstrated is the " f element", a nuclear Wolbachia insert involved in female sex determination in the terrestrial isopod Armadillidium vulgare . The f element represents an instance of bacteria-to-animal HGT that has occurred so recently that it was possible to infer the donor (feminizing Wolbachia closely related to the w VulC Wolbachia strain of A. vulgare ) and the mechanism of integration (a nearly complete genome inserted by micro-homology-mediated recombination). In this review, we summarize our current knowledge of the f element and discuss arising perspectives regarding female sex determination, unstable inheritance, population dynamics and the molecular evolution of the f element. Overall, the f element unifies three major areas in evolutionary biology: symbiosis, HGT and sex determination. Its characterization highlights the tremendous impact sex ratio distorters can have on the evolution of sex determination mechanisms and sex chromosomes in animals and plants.

The genetics and evolution of obligate reproductive parasitism in Trichogramma pretiosum infected with parthenogenesis-inducing Wolbachia

PubMed Central

Russell, J E; Stouthamer, R

2011-01-01

Parthenogenesis-inducing (PI) Wolbachia belong to a class of intracellular symbionts that distort the offspring sex ratio of their hosts toward a female bias. In many PI Wolbachia-infected species sex ratio distortion has reached its ultimate expression-fixation of infection and all-female populations. This is only possible with thelytokous PI symbionts as they provide an alternative form of reproduction and remove the requirement for males and sexual reproduction. Many populations fixed for PI Wolbachia infection have lost the ability to reproduce sexually, even when cured of the infection. We examine one such population in the species Trichogramma pretiosum. Through a series of backcrossing experiments with an uninfected Trichogramma pretiosum population we were able to show that the genetic basis for the loss of female sexual function could be explained by a dominant nuclear effect. Male sexual function had not been completely lost, though some deterioration of male sexual function was also evident when males from the infected population (created through antibiotic curing of infected females) were mated to uninfected females. We discuss the dynamics of sex ratio selection in PI Wolbachia-infected populations and the evolution of non-fertilizing mutations. PMID:20442735

The genetics and evolution of obligate reproductive parasitism in Trichogramma pretiosum infected with parthenogenesis-inducing Wolbachia.

PubMed

Russell, J E; Stouthamer, R

2011-01-01

Parthenogenesis-inducing (PI) Wolbachia belong to a class of intracellular symbionts that distort the offspring sex ratio of their hosts toward a female bias. In many PI Wolbachia-infected species sex ratio distortion has reached its ultimate expression-fixation of infection and all-female populations. This is only possible with thelytokous PI symbionts as they provide an alternative form of reproduction and remove the requirement for males and sexual reproduction. Many populations fixed for PI Wolbachia infection have lost the ability to reproduce sexually, even when cured of the infection. We examine one such population in the species Trichogramma pretiosum. Through a series of backcrossing experiments with an uninfected Trichogramma pretiosum population we were able to show that the genetic basis for the loss of female sexual function could be explained by a dominant nuclear effect. Male sexual function had not been completely lost, though some deterioration of male sexual function was also evident when males from the infected population (created through antibiotic curing of infected females) were mated to uninfected females. We discuss the dynamics of sex ratio selection in PI Wolbachia-infected populations and the evolution of non-fertilizing mutations.

Effect of naturally-occurring Wolbachia in Anopheles gambiae s.l. mosquitoes from Mali on Plasmodium falciparum malaria transmission

USDA-ARS?s Scientific Manuscript database

A novel naturally-occurring Wolbachia strain was identified in mosquitoes of the A. gambiae complex collected in the Malian villages of Dangassa and Kenieroba (wAnga-Mali). Phylogenetic analysis of the nucleotide sequence of two 16S rRNA regions showed that wAnga-Mali clusters with Wolbachia strains...

Field evaluation of the establishment potential of wMelPop Wolbachia in Australia and Vietnam for dengue control.

PubMed

Nguyen, Tran Hien; Nguyen, H Le; Nguyen, Thu Yen; Vu, Sinh Nam; Tran, Nhu Duong; Le, T N; Vien, Quang Mai; Bui, T C; Le, Huu Tho; Kutcher, Simon; Hurst, Tim P; Duong, T T H; Jeffery, Jason A L; Darbro, Jonathan M; Kay, B H; Iturbe-Ormaetxe, Iñaki; Popovici, Jean; Montgomery, Brian L; Turley, Andrew P; Zigterman, Flora; Cook, Helen; Cook, Peter E; Johnson, Petrina H; Ryan, Peter A; Paton, Chris J; Ritchie, Scott A; Simmons, Cameron P; O'Neill, Scott L; Hoffmann, Ary A

2015-10-28

Introduced Wolbachia bacteria can influence the susceptibility of Aedes aegypti mosquitoes to arboviral infections as well as having detrimental effects on host fitness. Previous field trials demonstrated that the wMel strain of Wolbachia effectively and durably invades Ae. aegypti populations. Here we report on trials of a second strain, wMelPop-PGYP Wolbachia, in field sites in northern Australia (Machans Beach and Babinda) and central Vietnam (Tri Nguyen, Hon Mieu Island), each with contrasting natural Ae. aegypti densities. Mosquitoes were released at the adult or pupal stages for different lengths of time at the sites depending on changes in Wolbachia frequency as assessed through PCR assays of material collected through Biogents-Sentinel (BG-S) traps and ovitraps. Adult numbers were also monitored through BG-S traps. Changes in Wolbachia frequency were compared across hamlets or house blocks. Releases of adult wMelPop-Ae. aegypti resulted in the transient invasion of wMelPop in all three field sites. Invasion at the Australian sites was heterogeneous, reflecting a slower rate of invasion in locations where background mosquito numbers were high. In contrast, invasion across Tri Nguyen was relatively uniform. After cessation of releases, the frequency of wMelPop declined in all sites, most rapidly in Babinda and Tri Nguyen. Within Machans Beach the rate of decrease varied among areas, and wMelPop was detected for several months in an area with a relatively low mosquito density. These findings highlight challenges associated with releasing Wolbachia-Ae. aegypti combinations with low fitness, albeit strong virus interference properties, as a means of sustainable control of dengue virus transmission.

Hindrances to bistable front propagation: application to Wolbachia invasion.

PubMed

Nadin, Grégoire; Strugarek, Martin; Vauchelet, Nicolas

2018-05-01

We study the biological situation when an invading population propagates and replaces an existing population with different characteristics. For instance, this may occur in the presence of a vertically transmitted infection causing a cytoplasmic effect similar to the Allee effect (e.g. Wolbachia in Aedes mosquitoes): the invading dynamics we model is bistable. We aim at quantifying the propagules (what does it take for an invasion to start?) and the invasive power (how far can an invading front go, and what can stop it?). We rigorously show that a heterogeneous environment inducing a strong enough population gradient can stop an invading front, which will converge in this case to a stable front. We characterize the critical population jump, and also prove the existence of unstable fronts above the stable (blocking) fronts. Being above the maximal unstable front enables an invading front to clear the obstacle and propagate further. We are particularly interested in the case of artificial Wolbachia infection, used as a tool to fight arboviruses.

Microsatellite and Wolbachia analysis in Rhagoletis cerasi natural populations: population structuring and multiple infections.

PubMed

Augustinos, Antonios A; Asimakopoulou, Anastasia K; Moraiti, Cleopatra A; Mavragani-Tsipidou, Penelope; Papadopoulos, Nikolaos T; Bourtzis, Kostas

2014-05-01

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.

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

The joint evolutionary histories of Wolbachia and mitochondria in Hypolimnas bolina.

PubMed

Charlat, Sylvain; Duplouy, Anne; Hornett, Emily A; Dyson, Emily A; Davies, Neil; Roderick, George K; Wedell, Nina; Hurst, Gregory D D

2009-03-24

The interaction between the Blue Moon butterfly, Hypolimnas bolina, and Wolbachia has attracted interest because of the high prevalence of male-killing achieved within the species, the ecological consequences of this high prevalence, the intensity of selection on the host to suppress the infection, and the presence of multiple Wolbachia infections inducing different phenotypes. We examined diversity in the co-inherited marker, mtDNA, and the partitioning of this between individuals of different infection status, as a means to investigate the population biology and evolutionary history of the Wolbachia infections. Part of the mitochondrial COI gene was sequenced from 298 individuals of known infection status revealing ten different haplotypes. Despite very strong biological evidence that the sample represents a single species, the ten haplotypes did not fall within a monophyletic clade within the Hypolimnas genus, with one haplotype differing by 5% from the other nine. There were strong associations between infection status and mtDNA haplotype. The presence of wBol1 infection in association with strongly divergent haplotypes prompted closer examination of wBol1 genetic variation. This revealed the existence of two cryptic subtypes, wBol1a and wBol1b. The wBol1a infection, by far the most common, was in strict association with the single divergent mtDNA haplotype. The wBol1b infection was found with two haplotypes that were also observed in uninfected specimens. Finally, the wBol2 infection was associated with a large diversity of mtDNA haplotypes, most often shared with uninfected sympatric butterflies. This data overall supports the hypothesis that high prevalence of male-killing Wolbachia (wBol1) in H. bolina is associated with very high transmission efficiency rather than regular horizontal transmission. It also suggests this infection has undergone a recent selective sweep and was introduced in this species through introgression. In contrast, the sharing of

Parasitism and mutualism in Wolbachia: what the phylogenomic trees can and cannot say.

PubMed

Bordenstein, Seth R; Paraskevopoulos, Charalampos; Dunning Hotopp, Julie C; Sapountzis, Panagiotis; Lo, Nathan; Bandi, Claudio; Tettelin, Hervé; Werren, John H; Bourtzis, Kostas

2009-01-01

Ecological and evolutionary theories predict that parasitism and mutualism are not fixed endpoints of the symbiotic spectrum. Rather, parasitism and mutualism may be host or environment dependent, induced by the same genetic machinery, and shifted due to selection. These models presume the existence of genetic or environmental variation that can spur incipient changes in symbiotic lifestyle. However, for obligate intracellular bacteria whose genomes are highly reduced, studies specify that discrete symbiotic associations can be evolutionarily stable for hundreds of millions of years. Wolbachia is an inherited obligate, intracellular infection of invertebrates containing taxa that act broadly as both parasites in arthropods and mutualists in certain roundworms. Here, we analyze the ancestry of mutualism and parasitism in Wolbachia and the evolutionary trajectory of this variation in symbiotic lifestyle with a comprehensive, phylogenomic analysis. Contrary to previous claims, we show unequivocally that the transition in lifestyle cannot be reconstructed with current methods due to long-branch attraction (LBA) artifacts of the distant Anaplasma and Ehrlichia outgroups. Despite the use of 1) site-heterogenous phylogenomic methods that can overcome systematic error, 2) a taxonomically rich set of taxa, and 3) statistical assessments of the genes, tree topologies, and models of evolution, we conclude that the LBA artifact is serious enough to afflict past and recent claims including the root lies in the middle of the Wolbachia mutualists and parasites. We show that different inference methods yield different results and high bootstrap support did not equal phylogenetic accuracy. Recombination was rare among this taxonomically diverse data set, indicating that elevated levels of recombination in Wolbachia are restricted to specific coinfecting groups. In conclusion, we attribute the inability to root the tree to rate heterogeneity between the ingroup and outgroup. Site

Wolbachia diversity and cytoplasmic incompatibility patterns in Culex pipiens populations in Turkey.

PubMed

Altinli, Mine; Gunay, Filiz; Alten, Bulent; Weill, Mylene; Sicard, Mathieu

2018-03-20

Wolbachia are maternally transmitted bacteria that can manipulate their hosts' reproduction causing cytoplasmic incompatibility (CI). CI is a sperm-egg incompatibility resulting in embryonic death. Due to this sterilising effect on mosquitoes, Wolbachia are considered for vector control strategies. Important vectors for arboviruses, filarial nematodes and avian malaria, mosquitoes of Culex pipiens complex are suitable for Wolbachia-based vector control. They are infected with Wolbachia wPip strains belonging to five genetically distinct groups (wPip-I to V) within the Wolbachia B supergroup. CI properties of wPip strongly correlate with this genetic diversity: mosquitoes infected with wPip strains from a different wPip group are more likely to be incompatible with each other. Turkey is a critical spot for vector-borne diseases due to its unique geographical position as a natural bridge between Asia, Europe and Africa. However, general wPip diversity, distribution and CI patterns in natural Cx. pipiens (s.l.) populations in the region are unknown. In this study, we first identified wPip diversity in Turkish Cx. pipiens (s.l.) populations, by assigning them to one of the five groups within wPip (wPip-Ito V). We further investigated CI properties between different wPip strains from this region. We showed a wPip fixation in Cx. pipiens (s.l.) populations in Turkey by analysing 753 samples from 59 sampling sites. Three wPip groups were detected in the region: wPip-I, wPip-II and wPip-IV. The most dominant group was wPip-II. While wPip-IV was restricted to only two locations, wPip-I and wPip-II had wider distributions. Individuals infected with wPip-II were found co-existing with individuals infected with wPip-I or wPip-IV in some sampling sites. Two mosquito isofemale lines harbouring either a wPip-I or a wPip-II strain were established from a population in northwestern Turkey. Reciprocal crosses between these lines showed that they were fully compatible with each

Removing the needle from the haystack: Enrichment of Wolbachia endosymbiont transcripts from host nematode RNA by Cappable-seq™.

PubMed

Luck, Ashley N; Slatko, Barton E; Foster, Jeremy M

2017-01-01

Efficient transcriptomic sequencing of microbial mRNA derived from host-microbe associations is often compromised by the much lower relative abundance of microbial RNA in the mixed total RNA sample. One solution to this problem is to perform extensive sequencing until an acceptable level of transcriptome coverage is obtained. More cost-effective methods include use of prokaryotic and/or eukaryotic rRNA depletion strategies, sometimes in conjunction with depletion of polyadenylated eukaryotic mRNA. Here, we report use of Cappable-seq™ to specifically enrich, in a single step, Wolbachia endobacterial mRNA transcripts from total RNA prepared from the parasitic filarial nematode, Brugia malayi. The obligate Wolbachia endosymbiont is a proven drug target for many human filarial infections, yet the precise nature of its symbiosis with the nematode host is poorly understood. Insightful analysis of the expression levels of Wolbachia genes predicted to underpin the mutualistic association and of known drug target genes at different life cycle stages or in response to drug treatments is typically challenged by low transcriptomic coverage. Cappable-seq resulted in up to ~ 5-fold increase in the number of reads mapping to Wolbachia. On average, coverage of Wolbachia transcripts from B. malayi microfilariae was enriched ~40-fold by Cappable-seq. Additionally, this method has an additional benefit of selectively removing abundant prokaryotic ribosomal RNAs.The deeper microbial transcriptome sequencing afforded by Cappable-seq facilitates more detailed characterization of gene expression levels of pathogens and symbionts present in animal tissues.

A lack of Wolbachia-specific DNA in samples from apollo butterfly (Parnassius apollo, Lepidoptera: Papilionidae) individuals with deformed or reduced wings.

PubMed

Łukasiewicz, Kinga; Sanak, Marek; Węgrzyn, Grzegorz

2016-05-01

Various insects contain maternally inherited endosymbiotic bacteria which can cause reproductive alterations, modulation of some physiological responses (like immunity, heat shock response, and oxidative stress response), and resistance to viral infections. In butterflies, Wolbachia sp. is the most frequent endosymbiont from this group, occurring in about 30 % of species tested to date. In this report, the presence of Wolbachia-specific DNA has been detected in apollo butterfly (Parnassius apollo). In the isolated population of this insect occurring in Pieniny National Park (Poland), malformed individuals with deformed or reduced wings appear with an exceptionally high frequency. Interestingly, while total DNA isolated from most (about 85 %) normal insects contained Wolbachia-specific sequences detected by PCR, such sequences were absent in a large fraction (70 %) of individuals with deformed wings and in all tested individuals with reduced wings. These results indicate for the first time the correlation between malformation of wings and the absence of Wolbachia sp. in insects. Although the lack of the endosymbiotic bacteria cannot be considered as the sole cause of the deformation or reduction of wings, one might suggest that Wolbachia sp. could play a protective role in the ontogenetic development of apollo butterfly.

Wolbachia wStri Blocks Zika Virus Growth at Two Independent Stages of Viral Replication.

PubMed

Schultz, M J; Tan, A L; Gray, C N; Isern, S; Michael, S F; Frydman, H M; Connor, J H

2018-05-22

Mosquito-transmitted viruses are spread globally and present a great risk to human health. Among the many approaches investigated to limit the diseases caused by these viruses are attempts to make mosquitos resistant to virus infection. Coinfection of mosquitos with the bacterium Wolbachia pipientis from supergroup A is a recent strategy employed to reduce the capacity for major vectors in the Aedes mosquito genus to transmit viruses, including dengue virus (DENV), Chikungunya virus (CHIKV), and Zika virus (ZIKV). Recently, a supergroup B Wolbachia w Stri, isolated from Laodelphax striatellus , was shown to inhibit multiple lineages of ZIKV in Aedes albopictus cells. Here, we show that w Stri blocks the growth of positive-sense RNA viruses DENV, CHIKV, ZIKV, and yellow fever virus by greater than 99.9%. w Stri presence did not affect the growth of the negative-sense RNA viruses LaCrosse virus or vesicular stomatitis virus. Investigation of the stages of the ZIKV life cycle inhibited by w Stri identified two distinct blocks in viral replication. We found a reduction of ZIKV entry into w Stri-infected cells. This was partially rescued by the addition of a cholesterol-lipid supplement. Independent of entry, transfected viral genome was unable to replicate in Wolbachia -infected cells. RNA transfection and metabolic labeling studies suggested that this replication defect is at the level of RNA translation, where we saw a 66% reduction in mosquito protein synthesis in w Stri-infected cells. This study's findings increase the potential for application of w Stri to block additional arboviruses and also identify specific blocks in viral infection caused by Wolbachia coinfection. IMPORTANCE Dengue, Zika, and yellow fever viruses are mosquito-transmitted diseases that have spread throughout the world, causing millions of infections and thousands of deaths each year. Existing programs that seek to contain these diseases through elimination of the mosquito population have so

"Good Passengers and Not Good Passengers:" Adolescent Drivers' Perceptions About Inattention and Peer Passengers.

PubMed

McDonald, Catherine C; Sommers, Marilyn S

The purpose of this qualitative focus group elicitation research study was to explore teen driver perceptions of peer passengers and driver inattention. We utilized focus groups for data collection and content analysis to analyze the data, both of which were guided by the theory of planned behavior. We conducted 7 focus groups with 30 teens, ages 16-18, licensed for ≤1year to examine attitudes, perceived behavioral control, and norms related to driving inattention and peer passengers. The sample was 50% male, mean age 17.39 (SD 0.52) with mean length of licensure 173.7days (SD 109.2). Three themes emerged: 1) "Good and not good" passengers; 2) Passengers and technology as harmful and helpful; and 3) The driver is in charge. While passengers can be a source of distraction, our participants also identified passenger behaviors that reduced risk, such as assistance with technology and guidance for directions. An understanding of teens' perceptions of peer passengers can contribute to the development of effective interventions targeting teen driver inattention. Nurses are well-positioned to contribute to these teen crash prevention efforts. Copyright © 2016 Elsevier Inc. All rights reserved.

Novel Wolbachia-transinfected Aedes aegypti mosquitoes possess diverse fitness and vector competence phenotypes.

PubMed

Fraser, Johanna E; De Bruyne, Jyotika Taneja; Iturbe-Ormaetxe, Iñaki; Stepnell, Justin; Burns, Rhiannon L; Flores, Heather A; O'Neill, Scott L

2017-12-01

Wolbachia pipientis from Drosophila melanogaster (wMel) is an endosymbiotic bacterium that restricts transmission of human pathogenic flaviviruses and alphaviruses, including dengue, Zika, and chikungunya viruses, when introduced into the mosquito vector Aedes aegypti. To date, wMel-infected Ae. aegypti have been released in field trials in 5 countries to evaluate the effectiveness of this strategy for disease control. Despite the success in establishing wMel-infected mosquitoes in wild populations, and the well-characterized antiviral capabilities of wMel, transinfecting different or additional Wolbachia strains into Ae. aegypti may improve disease impact, and perhaps more importantly, could provide a strategy to account for the possible evolution of resistant arboviruses. Here, we report the successful transinfection of Ae. aegypti with the Wolbachia strains wMelCS (D. melanogaster), wRi (D. simulans) and wPip (Culex quinquefasciatus) and assess the effects on Ae. aegypti fitness, cytoplasmic incompatibility, tissue tropism and pathogen blocking in a laboratory setting. The results demonstrate that wMelCS provides a similar degree of protection against dengue virus as wMel following an infectious blood meal, and significantly reduces viral RNA levels beyond that of wMel following a direct challenge with infectious virus in mosquitoes, with no additional fitness cost to the host. The protection provided by wRi is markedly weaker than that of wMelCS, consistent with previous characterisations of these lines in Drosophila, while wPip was found to substantially reduce the fitness of Ae. aegypti. Thus, we determine wMelCS as a key candidate for further testing in field-relevant fitness tests and viremic blood feeding challenges in a clinical setting to determine if it may represent an alternative Wolbachia strain with more desirable attributes than wMel for future field testing.

Wolbachia infection in Aedes aegypti mosquitoes alters blood meal excretion and delays oviposition without affecting trypsin activity.

PubMed

Pimenta de Oliveira, Sofia; Dantas de Oliveira, Caroline; Viana Sant'Anna, Mauricio Roberto; Carneiro Dutra, Heverton Leandro; Caragata, Eric Pearce; Moreira, Luciano Andrade

2017-08-01

Blood feeding in Aedes aegypti is essential for reproduction, but also permits the mosquito to act as a vector for key human pathogens such as the Zika and dengue viruses. Wolbachia pipientis is an endosymbiotic bacterium that can manipulate the biology of Aedes aegypti mosquitoes, making them less competent hosts for many pathogens. Yet while Wolbachia affects other aspects of host physiology, it is unclear whether it influences physiological processes associated with blood meal digestion. To that end, we examined the effects of wMel Wolbachia infection in Ae. aegypti, on survival post-blood feeding, blood meal excretion, rate of oviposition, expression levels of key genes involved in oogenesis, and activity levels of trypsin blood digestion enzymes. We observed that wMel infection altered the rate and duration of blood meal excretion, delayed the onset of oviposition and was associated with a greater number of eggs being laid later. wMel-infected Ae. aegypti also had lower levels of key yolk protein precursor genes necessary for oogenesis. However, all of these effects occurred without a change in trypsin activity. These results suggest that Wolbachia infection may disrupt normal metabolic processes associated with blood feeding and reproduction in Ae. aegypti. Copyright © 2017 Elsevier Ltd. All rights reserved.

Wolbachia wSinvictaA Infections in Natural Populations of the Fire Ant Solenopsis invicta: Testing for Phenotypic Effects

PubMed Central

Bouwma, Andrew M.; Shoemaker, DeWayne

2011-01-01

Wolbachia are intracellular bacteria that commonly infect many arthropods and some nematodes. In arthropods, these maternally transmitted bacteria often induce a variety of phenotypic effects to enhance their own spread within host populations. Wolbachia phenotypic effects generally either provide benefits to infected host females (cytoplasmic incompatibility, positive fitness effects) or bias host sex ratio in favor of females (male-killing, parthenogenesis, feminization), all of which increase the relative production of infected females in host populations. Wolbachia surveys have found infections to be exceedingly common in ants, but little is known at this juncture as to what phenotypic effects, if any, they induce in this group. Previous studies have demonstrated that individuals from native populations of the invasive fire ant Solenopsis invicta commonly harbor one or more of three Wolbachia variants. One of the variants, wSinvictaA, typically occurs at low prevalence in S. invicta populations, appears to have been transmitted horizontally into S. invicta three or more times, and has been lost repeatedly from host lineages over time. In order to determine the phenotypic effects and likely population dynamics of wSinvictaA infections in these ants, brood production patterns of newly mated fire ant queens were studied during simulated claustral founding and measured wSinvictaA transmission fidelity within mature single-queen families. No clear evidence was found for Wolbachia-mduced cytoplasmic incompatibility, significant fitness effects, or male-killing. Maternal transmission was perfect to both virgin queens and males. Possible mechanisms for how this variant could be maintained in host populations are discussed. PMID:21526927

Wolbachia wSinvictaA infections in natural populations of the fire ant Solenopsis invicta: testing for phenotypic effects.

PubMed

Bouwma, Andrew M; Shoemaker, Dewayne

2011-01-01

Wolbachia are intracellular bacteria that commonly infect many arthropods and some nematodes. In arthropods, these maternally transmitted bacteria often induce a variety of phenotypic effects to enhance their own spread within host populations. Wolbachia phenotypic effects generally either provide benefits to infected host females (cytoplasmic incompatibility, positive fitness effects) or bias host sex ratio in favor of females (male-killing, parthenogenesis, feminization), all of which increase the relative production of infected females in host populations. Wolbachia surveys have found infections to be exceedingly common in ants, but little is known at this juncture as to what phenotypic effects, if any, they induce in this group. Previous studies have demonstrated that individuals from native populations of the invasive fire ant Solenopsis invicta commonly harbor one or more of three Wolbachia variants. One of the variants, wSinvictaA, typically occurs at low prevalence in S. invicta populations, appears to have been transmitted horizontally into S. invicta three or more times, and has been lost repeatedly from host lineages over time. In order to determine the phenotypic effects and likely population dynamics of wSinvictaA infections in these ants, brood production patterns of newly mated fire ant queens were studied during simulated claustral founding and measured wSinvictaA transmission fidelity within mature single-queen families. No clear evidence was found for Wolbachia-induced cytoplasmic incompatibility, significant fitness effects, or male-killing. Maternal transmission was perfect to both virgin queens and males. Possible mechanisms for how this variant could be maintained in host populations are discussed.

Mito-nuclear genetic comparison in a Wolbachia infected weevil: insights on reproductive mode, infection age and evolutionary forces shaping genetic variation

PubMed Central

2010-01-01

Background Maternally inherited endosymbionts like Wolbachia pipientis are in linkage disequilibrium with the mtDNA of their hosts. Therefore, they can induce selective sweeps, decreasing genetic diversity over many generations. This sex ratio distorter, that is involved in the origin of parthenogenesis and other reproductive alterations, infects the parthenogenetic weevil Naupactus cervinus, a serious pest of ornamental and fruit plants. Results Molecular evolution analyses of mitochondrial (COI) and nuclear (ITS1) sequences from 309 individuals of Naupactus cervinus sampled over a broad range of its geographical distribution were carried out. Our results demonstrate lack of recombination in the nuclear fragment, non-random association between nuclear and mitochondrial genomes and the consequent coevolution of both genomes, being an indirect evidence of apomixis. This weevil is infected by a single Wolbachia strain, which could have caused a moderate bottleneck in the invaded population which survived the initial infection. Conclusions Clonal reproduction and Wolbachia infection induce the coevolution of bacterial, mitochondrial and nuclear genomes. The time elapsed since the Wolbachia invasion would have erased the traces of the demographic crash in the mtDNA, being the nuclear genome the only one that retained the signal of the bottleneck. The amount of genetic change accumulated in the mtDNA and the high prevalence of Wolbachia in all populations of N. cervinus agree with the hypothesis of an ancient infection. Wolbachia probably had great influence in shaping the genetic diversity of N. cervinus. However, it would have not caused the extinction of males, since sexual and asexual infected lineages coexisted until recent times. PMID:21050430

The joint evolutionary histories of Wolbachia and mitochondria in Hypolimnas bolina

PubMed Central

Charlat, Sylvain; Duplouy, Anne; Hornett, Emily A; Dyson, Emily A; Davies, Neil; Roderick, George K; Wedell, Nina; Hurst, Gregory DD

2009-01-01

Background The interaction between the Blue Moon butterfly, Hypolimnas bolina, and Wolbachia has attracted interest because of the high prevalence of male-killing achieved within the species, the ecological consequences of this high prevalence, the intensity of selection on the host to suppress the infection, and the presence of multiple Wolbachia infections inducing different phenotypes. We examined diversity in the co-inherited marker, mtDNA, and the partitioning of this between individuals of different infection status, as a means to investigate the population biology and evolutionary history of the Wolbachia infections. Results Part of the mitochondrial COI gene was sequenced from 298 individuals of known infection status revealing ten different haplotypes. Despite very strong biological evidence that the sample represents a single species, the ten haplotypes did not fall within a monophyletic clade within the Hypolimnas genus, with one haplotype differing by 5% from the other nine. There were strong associations between infection status and mtDNA haplotype. The presence of wBol1 infection in association with strongly divergent haplotypes prompted closer examination of wBol1 genetic variation. This revealed the existence of two cryptic subtypes, wBol1a and wBol1b. The wBol1a infection, by far the most common, was in strict association with the single divergent mtDNA haplotype. The wBol1b infection was found with two haplotypes that were also observed in uninfected specimens. Finally, the wBol2 infection was associated with a large diversity of mtDNA haplotypes, most often shared with uninfected sympatric butterflies. Conclusion This data overall supports the hypothesis that high prevalence of male-killing Wolbachia (wBol1) in H. bolina is associated with very high transmission efficiency rather than regular horizontal transmission. It also suggests this infection has undergone a recent selective sweep and was introduced in this species through introgression. In

Onchocerca parasites and Wolbachia endosymbionts: evaluation of a spectrum of antibiotic types for activity against Onchocerca gutturosa in vitro

PubMed Central

Townson, Simon; Tagboto, Senyo; McGarry, Helen F; Egerton, Gillian L; Taylor, Mark J

2006-01-01

Background The filarial parasites of major importance in humans contain the symbiotic bacterium Wolbachia and recent studies have shown that targeting of these bacteria with antibiotics results in a reduction in worm viability, development, embryogenesis, and survival. Doxycycline has been effective in human trials, but there is a need to develop drugs that can be given for shorter periods and to pregnant women and children. The World Health Organisation-approved assay to screen for anti-filarial activity in vitro uses male Onchocerca gutturosa, with effects being determined by worm motility and viability as measured by reduction of MTT to MTT formazan. Here we have used this system to screen antibiotics for anti-filarial activity. In addition we have determined the contribution of Wolbachia depletion to the MTT reduction assay. Methods Adult male O. gutturosa were cultured on a monkey kidney cell (LLCMK 2) feeder layer in 24-well plates with antibiotics and antibiotic combinations (6 to 10 worms per group). The macrofilaricide CGP 6140 (Amocarzine) was used as a positive control. Worm viability was assessed by two methods, (i) motility levels and (ii) MTT/formazan colorimetry. Worm motility was scored on a scale of 0 (immotile) to 10 (maximum) every 5 days up to 40 days. On day 40 worm viability was evaluated by MTT/formazan colorimetry, and results were expressed as a mean percentage reduction compared with untreated control values at day 40. To determine the contribution of Wolbachia to the MTT assay, the MTT formazan formation of an insect cell-line (C6/36) with or without insect Wolbachia infection and treated or untreated with tetracycline was compared. Results Antibiotics with known anti-Wolbachia activity were efficacious in this system. Rifampicin (5 × 10-5M) was the most effective anti-mycobacterial agent; clofazimine (1.25 × 10-5M and 3.13 × 10-6M) produced a gradual reduction in motility and by 40 days had reduced worm viability. The other anti

Evidence for a recent horizontal transmission and spatial spread of Wolbachia from endemic Rhagoletis cerasi (Diptera: Tephritidae) to invasive Rhagoletis cingulata in Europe.

PubMed

Schuler, Hannes; Bertheau, Coralie; Egan, Scott P; Feder, Jeffrey L; Riegler, Markus; Schlick-Steiner, Birgit C; Steiner, Florian M; Johannesen, Jes; Kern, Peter; Tuba, Katalin; Lakatos, Ferenc; Köppler, Kirsten; Arthofer, Wolfgang; Stauffer, Christian

2013-08-01

The widespread occurrence of Wolbachia in arthropods and nematodes suggests that this intracellular, maternally inherited endosymbiont has the ability to cross species boundaries. However, direct evidence for such a horizontal transmission of Wolbachia in nature is scarce. Here, we compare the well-characterized Wolbachia infection of the European cherry fruit fly, Rhagoletis cerasi, with that of the North American eastern cherry fruit fly, Rhagoletis cingulata, recently introduced to Europe. Molecular genetic analysis of Wolbachia based on multilocus sequence typing and the Wolbachia surface protein wsp showed that all R. cingulata individuals are infected with wCin2 identical to wCer2 in R. cerasi. In contrast, wCin1, a strain identical to wCer1 in R. cerasi, was present in several European populations of R. cingulata, but not in any individual from the United States. Surveys of R. cingulata from Germany and Hungary indicated that in some populations, the frequency of wCin1 increased significantly in just a few years with at least two independent horizontal transmission events. This is corroborated by the analysis of the mitochondrial cytochrome oxidase II gene that showed association of wCin1 with two distinct haplotypes in Germany, one of which is also infected with wCin1 in Hungary. In summary, our study provides strong evidence for a very recent inter-specific Wolbachia transmission with a subsequent spatial spread in field populations. © 2013 John Wiley & Sons Ltd.

The bacterium Wolbachia exploits host innate immunity to establish a symbiotic relationship with the dengue vector mosquito Aedes aegypti.

PubMed

Pan, Xiaoling; Pike, Andrew; Joshi, Deepak; Bian, Guowu; McFadden, Michael J; Lu, Peng; Liang, Xiao; Zhang, Fengrui; Raikhel, Alexander S; Xi, Zhiyong

2018-01-01

A host's immune system plays a central role in shaping the composition of the microbiota and, in return, resident microbes influence immune responses. Symbiotic associations of the maternally transmitted bacterium Wolbachia occur with a wide range of arthropods. It is, however, absent from the dengue and Zika vector mosquito Aedes aegypti in nature. When Wolbachia is artificially forced to form symbiosis with this new mosquito host, it boosts the basal immune response and enhances the mosquito's resistance to pathogens, including dengue, Zika virus and malaria parasites. The mechanisms involved in establishing a symbiotic relationship between Wolbachia and A. aegypti, and the long-term outcomes of this interaction, are not well understood. Here, we have demonstrated that both the immune deficiency (IMD) and Toll pathways are activated by the Wolbachia strain wAlbB upon its introduction into A. aegypti. Silencing the Toll and IMD pathways via RNA interference reduces the wAlbB load. Notably, wAlbB induces peptidoglycan recognition protein (PGRP)-LE expression in the carcass of A. aegypti, and its silencing results in a reduction of symbiont load. Using transgenic mosquitoes with stage-specific induction of the IMD and Toll pathways, we have shown that elevated wAlbB infection in these mosquitoes is maintained via maternal transmission. These results indicate that host innate immunity is utilized to establish and promote host-microbial symbiosis. Our results will facilitate a long-term projection of the stability of the Wolbachia-A. aegypti mosquito system that is being developed to control dengue and Zika virus transmission to humans.

Reproductive Manipulators in the Bark Beetle Pityogenes chalcographus (Coleoptera: Curculionidae)—The Role of Cardinium, Rickettsia, Spiroplasma, and Wolbachia

PubMed Central

Schebeck, Martin; Feldkirchner, Lukas; Marín, Belen; Krumböck, Susanne; Schuler, Hannes; Stauffer, Christian

2018-01-01

Abstract Heritable bacterial endosymbionts can alter the biology of numerous arthropods. They can influence the reproductive outcome of infected hosts, thus affecting the ecology and evolution of various arthropod species. The spruce bark beetle Pityogenes chalcographus (L.) (Coleoptera: Curculionidae: Scolytinae) was reported to express partial, unidirectional crossing incompatibilities among certain European populations. Knowledge on the background of these findings is lacking; however, bacterial endosymbionts have been assumed to manipulate the reproduction of this beetle. Previous work reported low-density and low-frequency Wolbachia infections of P. chalcographus but found it unlikely that this infection results in reproductive alterations. The aim of this study was to test the hypothesis of an endosymbiont-driven incompatibility, other than Wolbachia, reflected by an infection pattern on a wide geographic scale. We performed a polymerase chain reaction (PCR) screening of 226 individuals from 18 European populations for the presence of the endosymbionts Cardinium, Rickettsia, and Spiroplasma, and additionally screened these individuals for Wolbachia. Positive PCR products were sequenced to characterize these bacteria. Our study shows a low prevalence of these four endosymbionts in P. chalcographus. We detected a yet undescribed Spiroplasma strain in a single individual from Greece. This is the first time that this endosymbiont has been found in a bark beetle. Further, Wolbachia was detected in three beetles from two Scandinavian populations and two new Wolbachia strains were described. None of the individuals analyzed were infected with Cardinium and Rickettsia. The low prevalence of bacteria found here does not support the hypothesis of an endosymbiont-driven reproductive incompatibility in P. chalcographus. PMID:29771340

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

USDA-ARS?s Scientific Manuscript database

Wolbachia, the most widely studied endosymbiont in arthropods, is a target for biological control of mosquito-borne diseases (malaria and dengue virus), and antibiotic elimination of infectious filarial nematodes. We sequenced and analyzed the genome of a new strain (wPpe) in the plant-parasitic nem...

“Good passengers and not good passengers:” Adolescent drivers’ perceptions about inattention and peer passengers

PubMed Central

Sommers, Marilyn S.

2016-01-01

Purpose The purpose of this qualitative focus group elicitation research study was to explore teen driver perceptions of peer passengers and driver inattention. Design & Methods We utilized focus groups for data collection and content analysis to analyze the data, both of which were guided by the Theory of Planned Behavior. We conducted 7 focus groups with 30 teens, ages 16–18, licensed for ≤1 year to examine attitudes, perceived behavioral control, and norms related to driving inattention and peer passengers. Results The sample was 50% male, mean age 17.39 (sd 0.52) with mean length of licensure 173.7 days (sd 109.2). Three themes emerged: 1) “Good and not good” passengers; 2) Passengers and technology as harmful and helpful; and 3) The driver is in charge. Conclusions While passengers can be a source of distraction, our participants also identified passenger behaviors that reduced risk, such as assistance with technology and guidance for directions. Practical Implications An understanding of teens’ perceptions of peer passengers can contribute to the development of effective interventions targeting teen driver inattention. Nurses are well-positioned to contribute to these teen crash prevention efforts. PMID:27496828

Epidemiology of asexuality induced by the endosymbiotic Wolbachia across phytophagous wasp species: host plant specialization matters.

PubMed

Boivin, T; Henri, H; Vavre, F; Gidoin, C; Veber, P; Candau, J-N; Magnoux, E; Roques, A; Auger-Rozenberg, M-A

2014-05-01

Among eukaryotes, sexual reproduction is by far the most predominant mode of reproduction. However, some systems maintaining sexuality appear particularly labile and raise intriguing questions on the evolutionary routes to asexuality. Thelytokous parthenogenesis is a form of spontaneous loss of sexuality leading to strong distortion of sex ratio towards females and resulting from mutation, hybridization or infection by bacterial endosymbionts. We investigated whether ecological specialization is a likely mechanism of spread of thelytoky within insect communities. Focusing on the highly specialized genus Megastigmus (Hymenoptera: Torymidae), we first performed a large literature survey to examine the distribution of thelytoky in these wasps across their respective obligate host plant families. Second, we tested for thelytoky caused by endosymbionts by screening in 15 arrhenotokous and 10 thelytokous species for Wolbachia, Cardinium, Arsenophonus and Rickettsia endosymbionts and by performing antibiotic treatments. Finally, we performed phylogenetic reconstructions using multilocus sequence typing (MLST) to examine the evolution of endosymbiont-mediated thelytoky in Megastigmus and its possible connections to host plant specialization. We demonstrate that thelytoky evolved from ancestral arrhenotoky through the horizontal transmission and the fixation of the parthenogenesis-inducing Wolbachia. We find that ecological specialization in Wolbachia's hosts was probably a critical driving force for Wolbachia infection and spread of thelytoky, but also a constraint. Our work further reinforces the hypothesis that community structure of insects is a major driver of the epidemiology of endosymbionts and that competitive interactions among closely related species may facilitate their horizontal transmission. © 2014 John Wiley & Sons Ltd.

Minocycline as a re-purposed anti-Wolbachia macrofilaricide: superiority compared with doxycycline regimens in a murine infection model of human lymphatic filariasis

PubMed Central

Sharma, Raman; Jayoussi, Ghaith Al; Tyrer, Hayley E.; Gamble, Joanne; Hayward, Laura; Guimaraes, Ana F.; Davies, Jill; Waterhouse, David; Cook, Darren A. N.; Myhill, Laura J.; Clare, Rachel H.; Cassidy, Andrew; Steven, Andrew; Johnston, Kelly L.; Ford, Louise; Turner, Joseph D.; Ward, Stephen A.; Taylor, Mark J.

2016-01-01

Lymphatic filariasis and onchocerciasis are parasitic helminth diseases, which cause severe morbidities such as elephantiasis, skin disease and blindness, presenting a major public health burden in endemic communities. The anti-Wolbachia consortium (A·WOL: http://www.a-wol.com/) has identified a number of registered antibiotics that target the endosymbiotic bacterium, Wolbachia, delivering macrofilaricidal activity. Here we use pharmacokinetics/pharmacodynamics (PK/PD) analysis to rationally develop an anti-Wolbachia chemotherapy by linking drug exposure to pharmacological effect. We compare the pharmacokinetics and anti-Wolbachia efficacy in a murine Brugia malayi model of minocycline versus doxycycline. Doxycycline exhibits superior PK in comparison to minocycline resulting in a 3-fold greater exposure in SCID mice. Monte-Carlo simulations confirmed that a bi-daily 25–40 mg/Kg regimen is bioequivalent to a clinically effective 100–200 mg/day dose for these tetracyclines. Pharmacodynamic studies showed that minocycline depletes Wolbachia more effectively than doxycycline (99.51% vs. 90.35%) after 28 day 25 mg/Kg bid regimens with a more potent block in microfilarial production. PK/PD analysis predicts that minocycline would be expected to be 1.7 fold more effective than doxycycline in man despite lower exposure in our infection models. Our findings warrant onward clinical investigations to examine the clinical efficacy of minocycline treatment regimens against lymphatic filariasis and onchocerciasis. PMID:26996237

Evidence of natural Wolbachia infections and molecular identification of field populations of Culex pipiens complex (Diptera: Culicidae) mosquitoes in western Turkey.

PubMed

Morçiçek, Burçin; Taskin, Belgin Gocmen; Doğaç, Ersin; Doğaroğlu, Taylan; Taskin, Vatan

2018-06-01

Establishing reliable risk projection information about the distribution pattern of members of the Culex pipiens complex is of particular interest, as these mosquitoes are competent vectors for certain disease-causing pathogens. Wolbachia, a maternally inherited bacterial symbiont, are distributed in various arthropod species and can induce cytoplasmic incompatibility, i.e., reduced egg hatch, in certain crosses. It is being considered as a tool for population control of mosquito disease vectors. The Aegean region is characterized by highly populated, rural, and agricultural areas and is also on the route of the migratory birds. In this study, a fragment of the 658 bp of the mitochondrial cytochrome c oxidase subunit 1 (COI) gene, which includes the barcode region, was employed to differentiate Cx. pipiens complex species found in this region. Moreover, for the first time, the prevalence of Wolbachia endobacteria in these natural populations was examined using PCR amplification of a specific wsp gene. Our results revealed a widespread (more than 90%, n=121) presence of the highly efficient West Nile virus vector Cx. quinquefasciatus in the region. We also found that Wolbachia infection is widespread; the average prevalence was 62% in populations throughout the region. This study provided valuable information about the composition of Cx. pipiens complex mosquitoes and the prevalence of Wolbachia infection in these populations in the Aegean region. This information will be helpful in tracking mosquito-borne diseases and designing and implementing Wolbachia-based control strategies in the region. © 2018 The Society for Vector Ecology.

Deploying dengue-suppressing Wolbachia : Robust models predict slow but effective spatial spread in Aedes aegypti.

PubMed

Turelli, Michael; Barton, Nicholas H

2017-06-01

A novel strategy for controlling the spread of arboviral diseases such as dengue, Zika and chikungunya is to transform mosquito populations with virus-suppressing Wolbachia. In general, Wolbachia transinfected into mosquitoes induce fitness costs through lower viability or fecundity. These maternally inherited bacteria also produce a frequency-dependent advantage for infected females by inducing cytoplasmic incompatibility (CI), which kills the embryos produced by uninfected females mated to infected males. These competing effects, a frequency-dependent advantage and frequency-independent costs, produce bistable Wolbachia frequency dynamics. Above a threshold frequency, denoted pˆ, CI drives fitness-decreasing Wolbachia transinfections through local populations; but below pˆ, infection frequencies tend to decline to zero. If pˆ is not too high, CI also drives spatial spread once infections become established over sufficiently large areas. We illustrate how simple models provide testable predictions concerning the spatial and temporal dynamics of Wolbachia introductions, focusing on rate of spatial spread, the shape of spreading waves, and the conditions for initiating spread from local introductions. First, we consider the robustness of diffusion-based predictions to incorporating two important features of wMel-Aedes aegypti biology that may be inconsistent with the diffusion approximations, namely fast local dynamics induced by complete CI (i.e., all embryos produced from incompatible crosses die) and long-tailed, non-Gaussian dispersal. With complete CI, our numerical analyses show that long-tailed dispersal changes wave-width predictions only slightly; but it can significantly reduce wave speed relative to the diffusion prediction; it also allows smaller local introductions to initiate spatial spread. Second, we use approximations for pˆ and dispersal distances to predict the outcome of 2013 releases of wMel-infected Aedes aegypti in Cairns, Australia, Third

The effects of host age, host nuclear background and temperature on phenotypic effects of the virulent Wolbachia strain popcorn in Drosophila melanogaster.

PubMed Central

Reynolds, K Tracy; Thomson, Linda J; Hoffmann, Ary A

2003-01-01

Because of their obligate endosymbiotic nature, Wolbachia strains by necessity are defined by their phenotypic effects upon their host. Nevertheless, studies on the influence of host background and environmental conditions upon the manifestation of Wolbachia effects are relatively uncommon. Here we examine the behavior of the overreplicating Wolbachia strain popcorn in four different Drosophila melanogaster backgrounds at two temperatures. Unlike other strains of Wolbachia in Drosophila, popcorn has a major fitness impact upon its hosts. The rapid proliferation of popcorn causes cells to rupture, resulting in the premature death of adult hosts. Apart from this effect, we found that popcorn delayed development time, and host background influenced both this trait and the rate of mortality associated with infection. Temperature influenced the impact of popcorn upon host mortality, with no reduction in life span occurring in flies reared at 19 degrees. No effect upon fecundity was found. Contrary to earlier reports, popcorn induced high levels of incompatibility when young males were used in tests, and CI levels declined rapidly with male age. The population dynamics of popcorn-type infections will therefore depend on environmental temperature, host background, and the age structure of the population. PMID:12871912

The effects of host age, host nuclear background and temperature on phenotypic effects of the virulent Wolbachia strain popcorn in Drosophila melanogaster.

PubMed

Reynolds, K Tracy; Thomson, Linda J; Hoffmann, Ary A

2003-07-01

Because of their obligate endosymbiotic nature, Wolbachia strains by necessity are defined by their phenotypic effects upon their host. Nevertheless, studies on the influence of host background and environmental conditions upon the manifestation of Wolbachia effects are relatively uncommon. Here we examine the behavior of the overreplicating Wolbachia strain popcorn in four different Drosophila melanogaster backgrounds at two temperatures. Unlike other strains of Wolbachia in Drosophila, popcorn has a major fitness impact upon its hosts. The rapid proliferation of popcorn causes cells to rupture, resulting in the premature death of adult hosts. Apart from this effect, we found that popcorn delayed development time, and host background influenced both this trait and the rate of mortality associated with infection. Temperature influenced the impact of popcorn upon host mortality, with no reduction in life span occurring in flies reared at 19 degrees. No effect upon fecundity was found. Contrary to earlier reports, popcorn induced high levels of incompatibility when young males were used in tests, and CI levels declined rapidly with male age. The population dynamics of popcorn-type infections will therefore depend on environmental temperature, host background, and the age structure of the population.

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

USDA-ARS?s Scientific Manuscript database

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...

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. © 2016 The Authors. Molecular Ecology Published by John Wiley & Sons Ltd.

wFlu: Characterization and Evaluation of a Native Wolbachia from the Mosquito Aedes fluviatilis as a Potential Vector Control Agent

PubMed Central

Gonçalves, Daniela da Silva; Moreira, Luciano Andrade

2013-01-01

There is currently considerable interest and practical progress in using the endosymbiotic bacteria Wolbachia as a vector control agent for human vector-borne diseases. Such vector control strategies may require the introduction of multiple, different Wolbachia strains into target vector populations, necessitating the identification and characterization of appropriate endosymbiont variants. Here, we report preliminary characterization of wFlu, a native Wolbachia from the neotropical mosquito Aedes fluviatilis, and evaluate its potential as a vector control agent by confirming its ability to cause cytoplasmic incompatibility, and measuring its effect on three parameters determining host fitness (survival, fecundity and fertility), as well as vector competence (susceptibility) for pathogen infection. Using an aposymbiotic strain of Ae. fluviatilis cured of its native Wolbachia by antibiotic treatment, we show that in its natural host wFlu causes incomplete, but high levels of, unidirectional cytoplasmic incompatibility, has high rates of maternal transmission, and no detectable fitness costs, indicating a high capacity to rapidly spread through host populations. However, wFlu does not inhibit, and even enhances, oocyst infection with the avian malaria parasite Plasmodium gallinaceum. The stage- and sex-specific density of wFlu was relatively low, and with limited tissue distribution, consistent with the lack of virulence and pathogen interference/symbiont-mediated protection observed. Unexpectedly, the density of wFlu was also shown to be specifically-reduced in the ovaries after bloodfeeding Ae. fluviatilis. Overall, our observations indicate that the Wolbachia strain wFlu has the potential to be used as a vector control agent, and suggests that appreciable mutualistic coevolution has occurred between this endosymbiont and its natural host. Future work will be needed to determine whether wFlu has virulent host effects and/or exhibits pathogen interference when

NPS Transit System Passenger Boardings Study: Converting Ticket Sales to Passenger Boardings.

DOT National Transportation Integrated Search

2016-01-01

This report examines the reporting of passenger boardings (unlinked passenger trips) by NPS transit systems that use a ticket sales conversion methodology. By studying and validating the park units' passenger boarding methodology from converting tick...

Assessing the effect of extrinsic incubation period (EIP) prolongation in controlling dengue transmission with wolbachia-infected mosquito intervention

NASA Astrophysics Data System (ADS)

Putri, Y. E.; Rozi, S.; Tasman, H.; Aldila, D.

2017-03-01

A mathematical model of dengue disease transmission with involving Extrinsic Incubation Period (EIP) effect as a consequence of wolbachia introduction into mossquito population will be discussed in this article. Mathematical model analysis to find equilibrium points, basic reproductive ratio (ℛ0), and criteria of endemic occurrence which depend on some parameters were performed. From analytical result, we find that ℛ0 hold an important role to determine the existence and local stability of equilibrium points. From sensitivity analysis of ℛ0 and numerical simulation, we conclude that prolongation of EIP with wolbachia intervention succeed to reduce number of infected human and mosquito significantly.

Variation in Wolbachia effects on Aedes mosquitoes as a determinant of invasiveness and vectorial capacity.

PubMed

King, Jessica G; Souto-Maior, Caetano; Sartori, Larissa M; Maciel-de-Freitas, Rafael; Gomes, M Gabriela M

2018-04-16

Wolbachia has been introduced into Aedes aegypti mosquitoes to control the spread of arboviruses, such as dengue, chikungunya and Zika. Studies showed that certain Wolbachia strains (such as wMel) reduce replication of dengue viruses in the laboratory, prompting the release of mosquitoes carrying the bacterium into the field, where vectorial capacity can be realistically assessed in relation to native non-carriers. Here we apply a new analysis to two published datasets, and show that wMel increases the mean and the variance in Ae. aegypti susceptibility to dengue infection when introgressed into Brazil and Vietnam genetic backgrounds. In the absence of other processes, higher mean susceptibility should lead to enhanced viral transmission. The increase in variance, however, widens the basis for selection imposed by unexplored natural forces, retaining the potential for reducing transmission overall.

Combinations of registered drugs reduce treatment times required to deplete Wolbachia in the Litomosoides sigmodontis mouse model

PubMed Central

Specht, Sabine; Arriens, Sandra; Hübner, Marc P.; Klarmann-Schulz, Ute; Koschel, Marianne; Sternberg, Sonja; Martin, Coralie; Taylor, Mark J.; Hoerauf, Achim

2018-01-01

Filarial parasites can be targeted by antibiotic treatment due to their unique endosymbiotic relationship with Wolbachia bacteria. This finding has led to successful treatment strategies in both, human onchocerciasis and lymphatic filariasis. A 4–6 week treatment course using doxycycline results in long-term sterility and safe macrofilaricidal activity in humans. However, current treatment times and doxycycline contraindications in children and pregnant women preclude widespread administration of doxycycline in public health control programs; therefore, the search for shorter anti-wolbachial regimens is a focus of ongoing research. We have established an in vivo model for compound screening, using mice infected with Litomosoides sigmodontis. We could show that gold standard doxycycline treatment did not only deplete Wolbachia, it also resulted in a larval arrest. In this model, combinations of registered antibiotics were tested for their anti-wolbachial activity. Administration of rifamycins in combination with doxycycline for 7 days successfully depleted Wolbachia by > 2 log (>99% reduction) and thus resulted in a significant reduction of the treatment duration. Using a triple combination of a tetracycline (doxycycline or minocycline), a rifamycin and a fluoroquinolone (moxifloxacin) led to an even greater shortening of the treatment time. Testing all double combinations that could be derived from the triple combinations revealed that the combination of rifapentine (15mg/kg) and moxifloxacin (2 x 200mg/kg) showed the strongest reduction of treatment time in intraperitoneal and also oral administration routes. The rifapentine plus moxifloxacin combination was equivalent to the triple combination with additional doxycycline (>99% Wolbachia reduction). These investigations suggest that it is possible to shorten anti-wolbachial treatment times with combination treatments in order to achieve the target product profile (TPP) requirements for macrofilaricidal drugs of

Combinations of registered drugs reduce treatment times required to deplete Wolbachia in the Litomosoides sigmodontis mouse model.

PubMed

Specht, Sabine; Pfarr, Kenneth M; Arriens, Sandra; Hübner, Marc P; Klarmann-Schulz, Ute; Koschel, Marianne; Sternberg, Sonja; Martin, Coralie; Ford, Louise; Taylor, Mark J; Hoerauf, Achim

2018-01-01

Filarial parasites can be targeted by antibiotic treatment due to their unique endosymbiotic relationship with Wolbachia bacteria. This finding has led to successful treatment strategies in both, human onchocerciasis and lymphatic filariasis. A 4-6 week treatment course using doxycycline results in long-term sterility and safe macrofilaricidal activity in humans. However, current treatment times and doxycycline contraindications in children and pregnant women preclude widespread administration of doxycycline in public health control programs; therefore, the search for shorter anti-wolbachial regimens is a focus of ongoing research. We have established an in vivo model for compound screening, using mice infected with Litomosoides sigmodontis. We could show that gold standard doxycycline treatment did not only deplete Wolbachia, it also resulted in a larval arrest. In this model, combinations of registered antibiotics were tested for their anti-wolbachial activity. Administration of rifamycins in combination with doxycycline for 7 days successfully depleted Wolbachia by > 2 log (>99% reduction) and thus resulted in a significant reduction of the treatment duration. Using a triple combination of a tetracycline (doxycycline or minocycline), a rifamycin and a fluoroquinolone (moxifloxacin) led to an even greater shortening of the treatment time. Testing all double combinations that could be derived from the triple combinations revealed that the combination of rifapentine (15mg/kg) and moxifloxacin (2 x 200mg/kg) showed the strongest reduction of treatment time in intraperitoneal and also oral administration routes. The rifapentine plus moxifloxacin combination was equivalent to the triple combination with additional doxycycline (>99% Wolbachia reduction). These investigations suggest that it is possible to shorten anti-wolbachial treatment times with combination treatments in order to achieve the target product profile (TPP) requirements for macrofilaricidal drugs of no

"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,…

Using Near-Infrared Spectroscopy to Resolve the Species, Gender, Age, and the Presence of Wolbachia Infection in Laboratory-Reared Drosophila

PubMed Central

Aw, Wen C.; Dowell, Floyd E.; Ballard, J. William O.

2012-01-01

The aim of the study was to determine the accuracy of near-infrared spectroscopy (NIRS) in determining species, gender, age, and the presence of the common endosymbiont Wolbachia in laboratory-reared Drosophila. NIRS measures the absorption of light by organic molecules. Initially, a calibration model was developed for each study. An independent set with flies not involved in initial cross-validation was then used to validate the accuracy of each calibration model. Flies from the independent sets were correctly classified into Drosophila melanogaster and Drosophila simulans with 94% and 82% accuracy, respectively, whereas flies were successfully classified by gender with accuracy greater than 90%. In the age grading test, correlation plots of the actual and predicted age for males and females of D. melanogaster and D. simulans were shown to be overlapping between the adjacent age groups. It is, however, possible to predict the age of flies as less than 9 days of age with 62–88% accuracy and flies that are equal to or older than 9 days of age with 91–98% accuracy. Finally, we used NIRS to detect the presence of Wolbachia in flies. Flies from the independent sets were successfully identified as infected or not infected with Wolbachia with approximately 90% accuracy. These results suggest that NIRS has the potential to quantify the species, gender, and presence of Wolbachia in fly populations. However, additional optimization of the protocol may be necessary before the technique can reliably estimate fly age. PMID:22973543

Molecular genetics of the Wolbachia endosymbionts that infect the parasitoids of tephritid fruit flies.

USDA-ARS?s Scientific Manuscript database

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...

Two Strains of Male-Killing Wolbachia in a Ladybird, Coccinella undecimpunctata, from a Hot Climate

PubMed Central

Elnagdy, Sherif; Messing, Susan

2013-01-01

Ladybirds are a hot-spot for the invasion of male-killing bacteria. These maternally inherited endosymbionts cause the death of male host embryos, to the benefit of female sibling hosts and the bacteria that they contain. Previous studies have shown that high temperatures can eradicate male-killers from ladybirds, leaving the host free from infection. Here we report the discovery of two maternally inherited sex ratio distorters in populations of a coccinellid, Coccinella undecimpunctata, from a hot lowland region of the Middle East. DNA sequence analysis indicates that the male killing is the result of infection by Wolbachia, that the trait is tetracycline sensitive, and that two distinct strains of Wolbachia co-occur within one beetle population. We discuss the implications of these findings for theories of male-killing and suggest avenues for future field-work on this system. PMID:23349831

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.

The α-proteobacteria Wolbachia pipientis protein disulfide machinery has a regulatory mechanism absent in γ-proteobacteria.

PubMed

Walden, Patricia M; Halili, Maria A; Archbold, Julia K; Lindahl, Fredrik; Fairlie, David P; Inaba, Kenji; Martin, Jennifer L

2013-01-01

The α-proteobacterium Wolbachia pipientis infects more than 65% of insect species worldwide and manipulates the host reproductive machinery to enable its own survival. It can live in mutualistic relationships with hosts that cause human disease, including mosquitoes that carry the Dengue virus. Like many other bacteria, Wolbachia contains disulfide bond forming (Dsb) proteins that introduce disulfide bonds into secreted effector proteins. The genome of the Wolbachia strain wMel encodes two DsbA-like proteins sharing just 21% sequence identity to each other, α-DsbA1 and α-DsbA2, and an integral membrane protein, α-DsbB. α-DsbA1 and α-DsbA2 both have a Cys-X-X-Cys active site that, by analogy with Escherichia coli DsbA, would need to be oxidized to the disulfide form to serve as a disulfide bond donor toward substrate proteins. Here we show that the integral membrane protein α-DsbB oxidizes α-DsbA1, but not α-DsbA2. The interaction between α-DsbA1 and α-DsbB is very specific, involving four essential cysteines located in the two periplasmic loops of α-DsbB. In the electron flow cascade, oxidation of α-DsbA1 by α-DsbB is initiated by an oxidizing quinone cofactor that interacts with the cysteine pair in the first periplasmic loop. Oxidizing power is transferred to the second cysteine pair, which directly interacts with α-DsbA1. This reaction is inhibited by a non-catalytic disulfide present in α-DsbA1, conserved in other α-proteobacterial DsbAs but not in γ-proteobacterial DsbAs. This is the first characterization of the integral membrane protein α-DsbB from Wolbachia and reveals that the non-catalytic cysteines of α-DsbA1 regulate the redox relay system in cooperation with α-DsbB.

Cloning, Expression and Characterization of UDP-N-Acetylglucosamine Enolpyruvyl Transferase (MurA) from Wolbachia Endosymbiont of Human Lymphatic Filarial Parasite Brugia malayi

PubMed Central

Shahab, Mohd; Verma, Meenakshi; Pathak, Manisha; Mitra, Kalyan; Misra-Bhattacharya, Shailja

2014-01-01

Wolbachia, an endosymbiont of filarial nematode, is considered a promising target for treatment of lymphatic filariasis. Although functional characterization of the Wolbachia peptidoglycan assembly has not been fully explored, the Wolbachia genome provides evidence for coding all of the genes involved in lipid II biosynthesis, a part of peptidoglycan biosynthesis pathway. UDP-N-acetylglucosamine enolpyruvyl transferase (MurA) is one of the lipid II biosynthesis pathway enzymes and it has inevitably been recognized as an antibiotic target. In view of the vital role of MurA in bacterial viability and survival, MurA ortholog from Wolbachia endosymbiont of Brugia malayi (wBm-MurA) was cloned, expressed and purified for further molecular characterization. The enzyme kinetics and inhibition studies were undertaken using fosfomycin. wBm-MurA was found to be expressed in all the major life stages of B. malayi and was immunolocalized in Wolbachia within the microfilariae and female adults by the confocal microscopy. Sequence analysis suggests that the amino acids crucial for enzymatic activity are conserved. The purified wBm-MurA was shown to possess the EPSP synthase (3-phosphoshikimate 1-carboxyvinyltransferase) like activity at a broad pH range with optimal activity at pH 7.5 and 37°C temperature. The apparent affinity constant (K m) for the substrate UDP-N-acetylglucosamine was found to be 0.03149 mM and for phosphoenolpyruvate 0.009198 mM. The relative enzymatic activity was inhibited ∼2 fold in presence of fosfomycin. Superimposition of the wBm-MurA homology model with the structural model of Haemophilus influenzae (Hi-MurA) suggests binding of fosfomycin at the same active site. The findings suggest wBm-MurA to be a putative antifilarial drug target for screening of novel compounds. PMID:24941309

Air bags and passenger fatality according to passenger age and restraint use.

PubMed

Cummings, Peter; Koepsell, Thomas D; Rivara, Frederick P; McKnight, Barbara; Mack, Christopher

2002-09-01

Some children have been killed by air bags, leading to advice that young children should not sit in front of an active air bag. We conducted a case-control study to estimate the association of passenger air bag presence with death, according to passenger age and seat belt use. We used data from crashes on U.S. public roads in 1992 through 1998. Cases (N = 20,987) were front seat passengers who died, and controls (N = 69,277) were a sample of survivors. Among restrained passengers, the adjusted relative risk of death for those with a passenger air bag was 0.79 (95% confidence interval [CI] = 0.66-0.94); for children 12 years or younger, the adjusted relative risk was 1.04 (0.65-1.67) [corrected], and for adults 20 years or older it was 0.75 (0.62-0.91) [corrected]. Among unrestrained passengers, the adjusted relative risk was 1.03 (CI = 0.81-1.30); for children 12 years or younger the adjusted relative risk was 1.37 (0.84-2.21) [corrected], and for adults 20 years or older it was 0.96 (0.75-1.24). Passenger air bags may be a hazard to unrestrained children and of little benefit to unrestrained adults. Our results support the advice that children younger than 13 years should not sit in front of an active air bag.

Most influential FEMS publications.

PubMed

Prosser, James I; Cole, Jeff A; Nielsen, Jens; Bavoil, Patrik M; Häggblom, Max M

2014-05-01

A selection of influential FEMS publications to celebrate the 40th anniversary of FEMS. © 2014 Federation of European Microbiological Societies. Published by John Wiley & Sons Ltd. All rights reserved.

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

Code of Federal Regulations, 2012 CFR

2012-10-01

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

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

Code of Federal Regulations, 2013 CFR

2013-10-01

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

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

Code of Federal Regulations, 2010 CFR

2010-10-01

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

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

Code of Federal Regulations, 2014 CFR

2014-10-01

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

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

Code of Federal Regulations, 2011 CFR

2011-10-01

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

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

DOE Office of Scientific and Technical Information (OSTI.GOV)

Kurz, M.; Iturbe-Ormaetxe, I.; Jarrott, R.

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, bmore » = 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.« less

75 FR 26839 - Metrics and Standards for Intercity Passenger Rail Service under Section 207 of the Passenger...

Federal Register 2010, 2011, 2012, 2013, 2014

2010-05-12

... performance and service quality of intercity passenger train operations. In compliance with the statute, the FRA and Amtrak jointly drafted performance metrics and standards for intercity passenger rail service... and Standards for Intercity Passenger Rail Service under Section 207 of the Passenger Rail Investment...

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 or...

Reductions in deaths in frontal crashes among right front passengers in vehicles equipped with passenger air bags.

PubMed

Braver, E R; Ferguson, S A; Greene, M A; Lund, A K

1997-11-05

Virtually all new cars now are equipped with passenger air bags. Determining whether passenger air bags are saving lives is important, particularly because passenger air bags have caused some deaths among children and adults. To assess the effectiveness of passenger air bags in reducing the risk of death in frontal crashes for right front passengers. Air bags are designed to protect occupants in frontal crashes. Using Fatality Analysis Reporting System data for calendar years 1992 through 1995, the relative frequency of right front passenger deaths in frontal vs nonfrontal fatal crashes was compared for cars with dual air bags and for cars with driver-only air bags. Odds of right front passengers dying in frontal compared with nonfrontal fatal crashes were computed for 1992 through 1995 model year cars with dual air bags and for cars with driver-only air bags. Percentage reductions in right front passenger deaths in dual air bag vehicles were estimated. Right front passenger fatalities were 18% lower than expected in frontal crashes of cars with dual air bags and 11% lower in all crashes. An estimated 73 fewer than expected right front passengers died in 1992 through 1995 model cars with dual air bags during 1992 through 1995. The risk of frontal crash death for right front passengers in cars with dual air bags was reduced 14% among those reported to be using belts and 23% among belt nonusers. Children younger than 10 years in cars with dual air bags had a 34% increased risk of dying in frontal crashes. Passenger air bags were associated with substantial reductions in fatalities among right front passengers in frontal crashes. However, more children are being killed than are being saved by air bags. Immediate countermeasures to reduce the dangers of air bags to children and adults are suggested.

Decision-making in a bisexual line and a thelytokous Wolbachia-infected line of Trichogramma dendrolimi Matsumura (Hymenoptera: Trichogrammatidae) regarding behavior toward their hosts.

PubMed

Liu, Quan-Quan; Zhang, Tong-Shu; Li, Chun-Xue; Gu, Jun-Wen; Hou, Jie-Bin; Dong, Hui

2018-07-01

The use of thelytokous Wolbachia-infected Trichogramma (parasitic wasps) has long been considered as a way to enhance the efficacy of biocontrol. However, Wolbachia can affect the host physiology. We compared decision-making between bisexual and thelytokous Wolbachia-infected lines of Trichogramma dendrolimi Matsumura regarding behavior toward fresh and old eggs of Corcyra cephalonica at 25 ± 1 °C and 70 ± 5% relative humidity. The behavioral patterns and sequences of the two lines were basically the same. The durations of various behavioral patterns and values of fitness indicators of the bisexual line on fresh eggs were generally significantly shorter and better, respectively, than on old eggs, whereas the thelytokous line behaved similarly toward the two types of eggs, and differences in most fitness indicators between fresh and old eggs were not significant. On fresh eggs, the durations of various behaviors in the bisexual line were generally significantly shorter than in the thelytokous line and the fitness indicators were generally significantly better. Wolbachia affected the fitness of T. dendrolimi negatively. The potential of the thelytokous line as a biocontrol agent would not be as good as that of the bisexual line when decision-making only is considered. Therefore, further evaluations need to be carried out before the thelytokous line can be used in practical biocontrol. © 2018 Society of Chemical Industry. © 2018 Society of Chemical Industry.

19 CFR 4.50 - Passenger lists.

Code of Federal Regulations, 2010 CFR

2010-04-01

... 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 noncontiguous... 19 Customs Duties 1 2010-04-01 2010-04-01 false Passenger lists. 4.50 Section 4.50 Customs Duties...

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.

33 CFR 104.106 - Passenger access area.

Code of Federal Regulations, 2011 CFR

2011-07-01

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

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..., or cruise ship may designate areas within the vessel as passenger access areas. (b) A passenger...

33 CFR 104.106 - Passenger access area.

Code of Federal Regulations, 2013 CFR

2013-07-01

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

33 CFR 104.106 - Passenger access area.

Code of Federal Regulations, 2012 CFR

2012-07-01

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

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..., or cruise ship may designate areas within the vessel as passenger access areas. (b) A passenger...

Evolution of cooperation in a heterogeneous population with influential individuals

NASA Astrophysics Data System (ADS)

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

2012-02-01

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

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.

Global genetic variation in the Asian citrus psyllid, Diaphorina citri (Hemiptera: Liviidae) and the endosymbiont Wolbachia: links between Iran and the USA detected.

PubMed

Lashkari, Mohammadreza; Manzari, Shahab; Sahragard, Ahad; Malagnini, Valeria; Boykin, Laura M; Hosseini, Reza

2014-07-01

The Asian citrus psyllid, Diaphorina citri Kuwayama (Hemiptera: Liviidae), is one of the most serious pests of citrus in the world, because it transmits the pathogen that causes citrus greening disease. To determine genetic variation among geographic populations of D. citri, microsatellite markers, mitochondrial gene cytochrome oxidase I (mtCOI) and the Wolbachia-Diaphorina, wDi, gene wsp sequence data were used to characterize Iranian and Pakistani populations. Also, a Bayesian phylogenetic technique was utilized to elucidate the relationships among the sequences data in this study and all mtCOI and wsp sequence data available in GenBank and the Wolbachia database. Microsatellite markers revealed significant genetic differentiation among Iranian populations, as well as between Iranian and Pakistani populations (FST  = 0.0428, p < 0.01). Within Iran, the Sistan-Baluchestan population is significantly different from the Hormozgan (Fareghan) and Fars populations. By contrast, mtCOI data revealed two polymorphic sites separating the sequences from Iran and Pakistan. Global phylogenetic analyses showed that D. citri populations in Iran, India, Saudi Arabia, Brazil, Mexico, Florida and Texas (USA) are similar. Wolbachia, wDi, wsp sequences were similar among Iranian populations, but different between Iranian and Pakistani populations. The South West Asia (SWA) group is the most likely source of the introduced Iranian populations of D. citri. This assertion is also supported by the sequence similarity of the Wolbachia, wDi, strains from the Florida, USA and Iranian D. citri. These results should be considered when looking for biological controls in either country. © 2013 Society of Chemical Industry.

Passenger Value Structure Model

DOT National Transportation Integrated Search

1980-07-01

The objective of this research was to develop a model of the passenger's value structure which would reveal the role and importance of perceived security and other system characteristics on the passenger's evaluation and use of transit systems. The g...

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 includes...

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 includes...

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 includes...

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 includes...

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 includes...

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) manufactured...

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) manufactured...

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) manufactured...

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) manufactured...

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) manufactured...

14 CFR 91.1035 - Passenger awareness.

Code of Federal Regulations, 2011 CFR

2011-01-01

... orally briefed on— (1) Smoking: Each passenger must be briefed on when, where, and under what conditions smoking is prohibited. This briefing must include a statement, as appropriate, that the regulations require passenger compliance with lighted passenger information signs and no smoking placards, prohibit...

Passengers in containers

NASA Technical Reports Server (NTRS)

Tarkhanovskiy, V.

1977-01-01

A futuristic vision of future passenger and cargo transport is presented. To speed up lengthy transit operations, passengers would be accomodated in comfortable, compartment-like containers. Several diagrams show how such containers can be accomodated aboard an aircraft or a helicopter, on a truck, or in a railroad car. A system would result in great economy in both cost and time. Of particular importance is such a system for cargo traffic.

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

Code of Federal Regulations, 2011 CFR

2011-10-01

... 49 Transportation 4 2011-10-01 2011-10-01 false Periodic mechanical inspection of passenger cars and unpowered vehicles used in passenger trains. 238.307 Section 238.307 Transportation Other Regulations Relating to Transportation (Continued) FEDERAL RAILROAD ADMINISTRATION, DEPARTMENT OF TRANSPORTATION PASSENGER EQUIPMENT SAFETY STANDARDS...

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

Code of Federal Regulations, 2012 CFR

2012-10-01

... 49 Transportation 4 2012-10-01 2012-10-01 false Periodic mechanical inspection of passenger cars and unpowered vehicles used in passenger trains. 238.307 Section 238.307 Transportation Other Regulations Relating to Transportation (Continued) FEDERAL RAILROAD ADMINISTRATION, DEPARTMENT OF TRANSPORTATION PASSENGER EQUIPMENT SAFETY STANDARDS...

Genetic variation and potential coinfection of Wolbachia among widespread Asian citrus psyllid (Diaphorina citri Kuwayama) populations.

PubMed

Chu, Chia-Ching; Hoffmann, Mark; Braswell, W Evan; Pelz-Stelinski, Kirsten S

2017-12-29

Wolbachia can profoundly influence the survival, reproduction, and defenses of insect hosts. These interactions could potentially be harnessed for managing pests or insect-transmitted diseases. Diaphorina citri Kuwayama is a phloem-feeding pest capable of transmitting the putative causal agent of citrus greening, Candidatus Liberibacter asiaticus (CLas). Like many insects, D. citri is also infected with Wolbachia (wDi). Recent studies indicate that the relative abundance of wDi could be associated with the abundance of CLas, and that wDi may contribute to regulating expression of phage lytic cycle genes in CLas, suggesting the need for better understanding of wDi biology in general. This study investigated the genetic diversity of wDi among D. citri in populations spanning eleven countries and two U.S. territories. Six Wolbachia genes, wsp, coxA, fbpA, ftsZ, gatB, and hcpA, were sequenced and compared across samples. Two prevalent wDi strains were identified across the samples, and screening of clone libraries revealed possible coinfection of wDi strains in specific populations. D. citri mitochondrial cytochrome oxidase subunit I gene (mtCOI) were more divergent between D. citri populations that were infected with different wDi strains or had different infection statuses (single infection vs. coinfection). While we could not eliminate the possibility that maternal transmission may contribute to such patterns, it is also possible that wDi may induce cytoplasmic incompatibility in their host. These findings should contribute to the understanding of wDi population ecology, which may facilitate manipulation of this endosymbiont for management of citrus greening disease worldwide. © 2017 The Authors. Insect Science published by John Wiley & Sons Australia, Ltd on behalf of Institute of Zoology, Chinese Academy of Sciences.

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. Copyright © 2015 Elsevier Ltd. All rights reserved.

Intercity passenger rail : assessing the benefits of increased federal funding for Amtrak and high-speed passenger rail systems

DOT National Transportation Integrated Search

2001-03-21

The Rail Passenger Service Act of 1970 created Amtrak to provide intercity passenger rail service. Like other major national intercity passenger rail systems in the world, Amtrak has received substantial government support-nearly $24 billion for capi...

Parents as passengers during pediatric transport.

PubMed

Lewis, M M; Holditch-Davis, D; Brunssen, S

1997-01-01

The transport environment presents a unique setting in which the feasibility, advantages, and disadvantages of passengers accompanying a patient must be assessed carefully. The purpose of this study was to describe the current practice of including parents as passengers during pediatric interfacility transport. One-hundred-eighty-eight critical care transport programs in the United States responded to a voluntary mail survey, providing information about current policies, practices, and crew perceptions of the advantages and disadvantages of carrying parents as passengers. Extra seating for passengers was available in 96% of ambulances, 86% of fixed-wing aircraft, and 54% of helicopters used for pediatric transport. Parents traveled as passengers in all types of vehicles; most frequently in ambulances and fixed-wing aircraft. Twenty percent of helicopter programs allowed parent passengers on more than half of their pediatric transports in this vehicle. Advantages of parent passengers included emotional benefit for the parent and child, availability of parents for history and consent, good public relations, and having the parent present if the child dies. Disadvantages included potential parent anxiety, crew distraction, and space limitations. This study reflects the widely diverse policies, practices, and opinions relevant to this topic and confirms a need for further study.

Access improvement to aircraft passengers' hand luggage.

PubMed

Alberda, W; Kampinga, O; Kassels, R; van Kester, R; Noriega, J; Vink, P

2015-01-01

Efficient use of space and passenger comfort in aircraft interiors are major issues. There is not much research available about the flying experience regarding passengers' personal belongings. The objective of this study is to explore concepts within the current aircraft seats which improve the passenger experience related to their personal belongings like wallets, mobile phones and laptops. Through on-site observations, interviews and online questionnaires, data regarding the number of personal belongings taken into the airplane and opinions about access to hand luggage were gathered. These data were used to develop different concepts to optimize the aircraft interior, which were evaluated by passengers. Almost every passenger carries a phone (88%), wallet (94%), travel documents (98%) and keys (76%) with them and they like to have these stored close by. Passengers rate the concept that provides integrated storage in the tray table of the aircraft seat the best. Extra storage possibility in the table-tray seems a promising solution according to the passengers.

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 any...

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 any...

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 any...

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 any...

Network structure of subway passenger flows

NASA Astrophysics Data System (ADS)

Xu, Q.; Mao, B. H.; Bai, Y.

2016-03-01

The results of transportation infrastructure network analyses have been used to analyze complex networks in a topological context. However, most modeling approaches, including those based on complex network theory, do not fully account for real-life traffic patterns and may provide an incomplete view of network functions. This study utilizes trip data obtained from the Beijing Subway System to characterize individual passenger movement patterns. A directed weighted passenger flow network was constructed from the subway infrastructure network topology by incorporating trip data. The passenger flow networks exhibit several properties that can be characterized by power-law distributions based on flow size, and log-logistic distributions based on the fraction of boarding and departing passengers. The study also characterizes the temporal patterns of in-transit and waiting passengers and provides a hierarchical clustering structure for passenger flows. This hierarchical flow organization varies in the spatial domain. Ten cluster groups were identified, indicating a hierarchical urban polycentric structure composed of large concentrated flows at urban activity centers. These empirical findings provide insights regarding urban human mobility patterns within a large subway network.

Teen Drivers' Perceptions of Their Peer Passengers: Qualitative Study.

PubMed

Ehsani, Johnathon P; Haynie, Denise L; Luthers, Christina; Perlus, Jessamyn; Gerber, Eli; Ouimet, Marie Claude; Klauer, Sheila G; Simons-Morton, Bruce

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. 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. 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. 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.

Enforcement of the child passenger protection law

DOT National Transportation Integrated Search

1980-10-01

This report describes the Tennessee law enforcement agencies and their role in the enforcement of the child passenger protection law. Chapter I introduces the importance of protecting child passengers in motor vehicles, describes the child passenger ...

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

USDA-ARS?s Scientific Manuscript database

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...

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.

Securing America’s Passenger-Rail Systems

DTIC Science & Technology

2007-01-01

around the world highlight the vulnerability of rail travel and the importance of rail security for these passengers. The use of passenger rail and...take to U.S. railways. Recent attacks on passenger-rail systems around the world highlight the vulnerability of rail travel and the importance of rail...rails (Boardman, 2005), making more than 3.5 billion trips (APTA, 2006).1 And these estimates do not count the passengers traveling on the National

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 Certificate...

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 Certificate...

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 Certificate...

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 Certificate...

Ranking influential spreaders is an ill-defined problem

NASA Astrophysics Data System (ADS)

Gu, Jain; Lee, Sungmin; Saramäki, Jari; Holme, Petter

2017-06-01

Finding influential spreaders of information and disease in networks is an important theoretical problem, and one of considerable recent interest. It has been almost exclusively formulated as a node-ranking problem —methods for identifying influential spreaders output a ranking of the nodes. In this work, we show that such a greedy heuristic does not necessarily work: the set of most influential nodes depends on the number of nodes in the set. Therefore, the set of n most important nodes to vaccinate does not need to have any node in common with the set of n + 1 most important nodes. We propose a method for quantifying the extent and impact of this phenomenon. By this method, we show that it is a common phenomenon in both empirical and model networks.

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.

Lutzomyia Sand Fly Diversity and Rates of Infection by Wolbachia and an Exotic Leishmania Species on Barro Colorado Island, Panama

PubMed Central

Azpurua, Jorge; De La Cruz, Dianne; Valderama, Anayansi; Windsor, Donald

2010-01-01

Background Sand flies (Diptera, Psychodidae, Phlebotominae) in the genus Lutzomyia are the predominant vectors of the protozoan disease leishmaniasis in the New World. Within the watershed of the Panama Canal, the cutaneous form of leishmaniasis is a continuous health threat for residents, tourists and members of an international research community. Here we report the results of screening a tropical forest assemblage of sand fly species for infection by both Leishmania and a microbe that can potentially serve in vector population control, the cytoplasmically transmitted rickettsia, Wolbachia pipientis. Knowing accurately which Lutzomyia species are present, what their evolutionary relationships are, and how they are infected by strains of both Leishmania and Wolbachia is of critical value for building strategies to mitigate the impact of this disease in humans. Methodology and Findings We collected, sorted and then used DNA sequences to determine the diversity and probable phylogenetic relationships of the Phlebotominae occurring in the understory of Barro Colorado Island in the Republic of Panama. Sequence from CO1, the DNA barcoding gene, supported 18 morphology-based species determinations while revealing the presence of two possible “cryptic” species, one (Lu. sp. nr vespertilionis) within the Vespertilionis group, the other (Lu. gomezi) within the Lutzomyia-cruciata series. Using ITS-1 and “minicircle” primers we detected Leishmania DNA in 43.3% of Lu. trapidoi, 26.3% of Lu. gomezi individuals and in 0% of the other 18 sand fly species. Identical ITS-1 sequence was obtained from the Leishmania infecting Lu. trapidoi and Lu. gomezi, sequence which was 93% similar to Leishmania (viannia) naiffi in GenBank, a species previously unknown in Panama, but recognized as a type of cutaneous leishmaniasis vectored broadly across northern and central South America. Distinct strains of the intracellular bacterium Wolbachia were detected in three of 20 sand fly

The impact of temperature and Wolbachia infection on vector competence of potential dengue vectors Aedes aegypti and Aedes albopictus in the transmission of dengue virus serotype 1 in southern Taiwan.

PubMed

Tsai, Cheng-Hui; Chen, Tien-Huang; Lin, Cheo; Shu, Pei-Yun; Su, Chien-Ling; Teng, Hwa-Jen

2017-11-07

We evaluated the impact of temperature and Wolbachia infection on vector competence of the local Aedes aegypti and Ae. albopictus populations of southern Taiwan in the laboratory. After oral infection with dengue serotype 1 virus (DENV-1), female mosquitoes were incubated at temperatures of 10, 16, 22, 28 and 34 °C. Subsequently, salivary gland, head, and thorax-abdomen samples were analyzed for their virus titer at 0, 5, 10, 15, 20, 25 and 30 days post-infection (dpi) by real-time RT-PCR. The results showed that Ae. aegypti survived significantly longer and that dengue viral genome levels in the thorax-abdomen (10 3.25 ± 0.53 -10 4.09 ± 0.71 PFU equivalents/ml) and salivary gland samples (10 2.67 ± 0.33 -10 3.89 ± 0.58 PFU equivalents/ml) were significantly higher at high temperature (28-34 °C). The survival of Ae. albopictus was significantly better at 16 or 28 °C, but the virus titers from thorax-abdomen (10 0.70 -10 2.39 ± 1.31 PFU equivalents/ml) and salivary gland samples (10 0.12 ± 0.05 -10 1.51 ± 0.31 PFU equivalents/ml) were significantly higher at 22-28 °C. Within viable temperature ranges, the viruses were detectable after 10 dpi in salivary glands and head tissues in Ae. aegypti and after 5-10 dpi in Ae. albopictus. Vector competence was measured in Ae. albopictus with and without Wolbachia at 28 °C. Wolbachia-infected mosquitoes survived significantly better and carried lower virus titers than Wolbachia-free mosquitoes. Wolbachia coinfections (92.8-97.2%) with wAlbA and wAlbB strains were commonly found in a wild population of Ae. albopictus. In southern Taiwan, Ae. aegypti is the main vector of dengue and Ae. albopictus has a non-significant role in the transmission of dengue virus due to the high prevalence of Wolbachia infection in the local mosquito population of southern Taiwan.

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.

Ontology Design of Influential People Identification Using Centrality

NASA Astrophysics Data System (ADS)

Maulana Awangga, Rolly; Yusril, Muhammad; Setyawan, Helmi

2018-04-01

Identifying influential people as a node in a graph theory commonly calculated by social network analysis. The social network data has the user as node and edge as relation forming a friend relation graph. This research is conducting different meaning of every nodes relation in the social network. Ontology was perfect match science to describe the social network data as conceptual and domain. Ontology gives essential relationship in a social network more than a current graph. Ontology proposed as a standard for knowledge representation for the semantic web by World Wide Web Consortium. The formal data representation use Resource Description Framework (RDF) and Web Ontology Language (OWL) which is strategic for Open Knowledge-Based website data. Ontology used in the semantic description for a relationship in the social network, it is open to developing semantic based relationship ontology by adding and modifying various and different relationship to have influential people as a conclusion. This research proposes a model using OWL and RDF for influential people identification in the social network. The study use degree centrality, between ness centrality, and closeness centrality measurement for data validation. As a conclusion, influential people identification in Facebook can use proposed Ontology model in the Group, Photos, Photo Tag, Friends, Events and Works data.

Identifying influential spreaders in complex networks based on kshell hybrid method

NASA Astrophysics Data System (ADS)

Namtirtha, Amrita; Dutta, Animesh; Dutta, Biswanath

2018-06-01

Influential spreaders are the key players in maximizing or controlling the spreading in a complex network. Identifying the influential spreaders using kshell decomposition method has become very popular in the recent time. In the literature, the core nodes i.e. with the largest kshell index of a network are considered as the most influential spreaders. We have studied the kshell method and spreading dynamics of nodes using Susceptible-Infected-Recovered (SIR) epidemic model to understand the behavior of influential spreaders in terms of its topological location in the network. From the study, we have found that every node in the core area is not the most influential spreader. Even a strategically placed lower shell node can also be a most influential spreader. Moreover, the core area can also be situated at the periphery of the network. The existing indexing methods are only designed to identify the most influential spreaders from core nodes and not from lower shells. In this work, we propose a kshell hybrid method to identify highly influential spreaders not only from the core but also from lower shells. The proposed method comprises the parameters such as kshell power, node's degree, contact distance, and many levels of neighbors' influence potential. The proposed method is evaluated using nine real world network datasets. In terms of the spreading dynamics, the experimental results show the superiority of the proposed method over the other existing indexing methods such as the kshell method, the neighborhood coreness centrality, the mixed degree decomposition, etc. Furthermore, the proposed method can also be applied to large-scale networks by considering the three levels of neighbors' influence potential.

The impact of Wolbachia, male age and mating history on cytoplasmic incompatibility and sperm transfer in Drosophila simulans.

PubMed

Awrahman, Z A; Champion de Crespigny, F; Wedell, N

2014-01-01

Most insects harbour a variety of maternally inherited endosymbionts, the most widespread being Wolbachia pipientis that commonly induce cytoplasmic incompatibility (CI) and reduced hatching success in crosses between infected males and uninfected females. High temperature and increasing male age are known to reduce the level of CI in a variety of insects. In Drosophila simulans, infected males have been shown to mate at a higher rate than uninfected males. By examining the impact of mating rate independent of age, this study investigates whether a high mating rate confers an advantage to infected males through restoring their compatibility with uninfected females over and above the effect of age. The impact of Wolbachia infection, male mating rate and age on the number of sperm transferred to females during copulation and how it relates to CI expression was also assessed. As predicted, we found that reproductive compatibility was restored faster in males that mate at higher rate than that of low mating and virgin males, and that the effect of mating history was over and above the effect of male age. Nonvirgin infected males transferred fewer sperm than uninfected males during copulation, and mating at a high rate resulted in the transfer of fewer sperm per mating irrespective of infection status. These results indicate that the advantage to infected males of mating at a high rate is through restoration of reproductive compatibility with uninfected females, whereas uninfected males appear to trade off the number of sperm transferred per mating with female encounter rate and success in sperm competition. This study highlights the importance Wolbachia may play in sexual selection by affecting male reproductive strategies. © 2013 The Authors. Journal of Evolutionary Biology © 2013 European Society For Evolutionary Biology.

The influence of passenger car front shape on pedestrian injury risk observed from German in-depth accident data.

PubMed

Li, Guibing; Lyons, Mathew; Wang, Bingyu; Yang, Jikuang; Otte, Dietmar; Simms, Ciaran

2017-04-01

odds increase by 74% for a 10% increase in NBLEH. These findings suggest that passenger car bumpers should support the lower leg with a low and flat lower bumper and even contact up to the femur area with a high upper bumper which extends above the knee to protect the pedestrian's leg. A low passenger car bonnet leading edge helps to reduce femur/pelvis injury risk. The passenger car front shape parameters are less influential than impact speed and pedestrian age for pedestrian injury risk. Copyright © 2017 Elsevier Ltd. All rights reserved.

Using near-infrared spectroscopy to resolve the species, gender, age, and the presence of Wolbachia infection in laboratory-reared Drosophila

USDA-ARS?s Scientific Manuscript database

The aim of the study was to determine the accuracy of near-infrared spectroscopy (NIRS) in determining species, gender, age and the presence of the common endosymbiont Wolbachia in laboratory reared Drosophila. NIRS measures absorption of light by organic molecules. Initially, a calibration model wa...

49 CFR 223.15 - Requirements for existing passenger cars.

Code of Federal Regulations, 2010 CFR

2010-10-01

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

49 CFR 223.15 - Requirements for existing passenger cars.

Code of Federal Regulations, 2014 CFR

2014-10-01

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

49 CFR 223.15 - Requirements for existing passenger cars.

Code of Federal Regulations, 2011 CFR

2011-10-01

... 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 CABOOSES Specific Requirements § 223.15 Requirements for existing passenger cars. (a) Passenger cars built or...

49 CFR 223.15 - Requirements for existing passenger cars.

Code of Federal Regulations, 2012 CFR

2012-10-01

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

49 CFR 223.15 - Requirements for existing passenger cars.

Code of Federal Regulations, 2013 CFR

2013-10-01

... 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 CABOOSES Specific Requirements § 223.15 Requirements for existing passenger cars. (a) Passenger cars built or...

The Effect of Passengers on Teen Driver Behavior

DOT National Transportation Integrated Search

2012-04-01

A number of studies have shown that passengers substantially increase the risk of crashes for young, novice drivers. This increased risk may result from distractions that young passengers create for drivers. Alternatively, the presence of passengers ...

Optimal boarding method for airline passengers

DOE Office of Scientific and Technical Information (OSTI.GOV)

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 andmore » discuss practical modifications to the optimal. Finally, I mention some of the benefits that could come from implementing an improved passenger boarding scheme.« less

49 CFR 239.103 - Passenger train emergency simulations.

Code of Federal Regulations, 2014 CFR

2014-10-01

... 49 Transportation 4 2014-10-01 2014-10-01 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 execute...

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 execute...

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 execute...

49 CFR 239.103 - Passenger train emergency simulations.

Code of Federal Regulations, 2013 CFR

2013-10-01

... 49 Transportation 4 2013-10-01 2013-10-01 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 execute...

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.

Passenger Travel Facts and Figures 2014

DOT National Transportation Integrated Search

2014-12-17

Passenger Travel Facts and Figures 2014 is a snapshot of the characteristics and trends of personal travel in the United States; the network over which passenger travel takes place; and the related economic, safety, and environmental implications. An...

Passenger Travel Facts and Figures 2016

DOT National Transportation Integrated Search

2016-08-10

Passenger Travel Facts and Figures 2016 is a snapshot of the characteristics and trends of personal travel in the United States; the network over which passenger travel takes place; and the related economic, safety, and environmental aspects of passe...

Passenger Travel Facts and Figures 2015

DOT National Transportation Integrated Search

2015-09-16

Passenger Travel Facts and Figures 2015 is a snapshot of the characteristics and trends of personal travel in the United States; the network over which passenger travel takes place; and the related economic, safety, and environmental aspects of passe...

14 CFR 19-7 - Passenger origin-destination survey.

Code of Federal Regulations, 2012 CFR

2012-01-01

... 14 Aeronautics and Space 4 2012-01-01 2012-01-01 false Passenger origin-destination survey. Sec... AIR CARRIERS Operating Statistics Classifications Sec. 19-7 Passenger origin-destination survey. (a... carriers) shall participate in a Passenger Origin-Destination (O & D) Survey covering domestic and...

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

Population structure in relation to host-plant ecology and Wolbachia infestation in the comma butterfly.

PubMed

Kodandaramaiah, U; Weingartner, E; Janz, N; Dalén, L; Nylin, S

2011-10-01

Experimental work on Polygonia c-album, a temperate polyphagous butterfly species, has shown that Swedish, Belgian, Norwegian and Estonian females are generalists with respect to host-plant preference, whereas females from UK and Spain are specialized on Urticaceae. Female preference is known to have a strong genetic component. We test whether the specialist and generalist populations form respective genetic clusters using data from mitochondrial sequences and 10 microsatellite loci. Results do not support this hypothesis, suggesting that the specialist and generalist traits have evolved more than once independently. Mitochondrial DNA variation suggests a rapid expansion scenario, with a single widespread haplotype occurring in high frequency, whereas microsatellite data indicate strong differentiation of the Moroccan population. Based on a comparison of polymorphism in the mitochondrial data and sequences from a nuclear gene, we show that the diversity in the former is significantly less than that expected under neutral evolution. Furthermore, we found that almost all butterfly samples were infected with a single strain of Wolbachia, a maternally inherited bacterium. We reason that indirect selection on the mitochondrial genome mediated by a recent sweep of Wolbachia infection has depleted variability in the mitochondrial sequences. We also surmise that P. c-album could have expanded out of a single glacial refugium and colonized Morocco recently. © 2011 The Authors. Journal of Evolutionary Biology © 2011 European Society For Evolutionary Biology.

[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.

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

Infection with a Virulent Strain of Wolbachia Disrupts Genome Wide-Patterns of Cytosine Methylation in the Mosquito Aedes aegypti

PubMed Central

Ye, Yixin H.; Woolfit, Megan; Huttley, Gavin A.; Rancès, Edwige; Caragata, Eric P.; Popovici, Jean; O'Neill, Scott L.; McGraw, Elizabeth A.

2013-01-01

Background Cytosine methylation is one of several reversible epigenetic modifications of DNA that allow a greater flexibility in the relationship between genotype and phenotype. Methylation in the simplest models dampens gene expression by modifying regions of DNA critical for transcription factor binding. The capacity to methylate DNA is variable in the insects due to diverse histories of gene loss and duplication of DNA methylases. Mosquitoes like Drosophila melanogaster possess only a single methylase, DNMT2. Description Here we characterise the methylome of the mosquito Aedes aegypti and examine its relationship to transcription and test the effects of infection with a virulent strain of the endosymbiont Wolbachia on the stability of methylation patterns. Conclusion We see that methylation in the A. aegypti genome is associated with reduced transcription and is most common in the promoters of genes relating to regulation of transcription and metabolism. Similar gene classes are also methylated in aphids and honeybees, suggesting either conservation or convergence of methylation patterns. In addition to this evidence of evolutionary stability, we also show that infection with the virulent wMelPop Wolbachia strain induces additional methylation and demethylation events in the genome. While most of these changes seem random with respect to gene function and have no detected effect on transcription, there does appear to be enrichment of genes associated with membrane function. Given that Wolbachia lives within a membrane-bound vacuole of host origin and retains a large number of genes for transporting host amino acids, inorganic ions and ATP despite a severely reduced genome, these changes might represent an evolved strategy for manipulating the host environments for its own gain. Testing for a direct link between these methylation changes and expression, however, will require study across a broader range of developmental stages and tissues with methods that detect

Influential Observations in Principal Factor Analysis.

ERIC Educational Resources Information Center

Tanaka, Yutaka; Odaka, Yoshimasa

1989-01-01

A method is proposed for detecting influential observations in iterative principal factor analysis. Theoretical influence functions are derived for two components of the common variance decomposition. The major mathematical tool is the influence function derived by Tanaka (1988). (SLD)

Passenger Flow Estimation and Characteristics Expansion

DOT National Transportation Integrated Search

2016-04-01

Mark R. McCord (ORCID ID 0000-0002-6293-3143) The objectives of this study are to investigate the estimation of bus passenger boarding-to-alighting (B2A) flows using Automatic Passenger Count (APC) and Automatic Fare Collection (AFC) (fare-box) data ...

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 cognizant...

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 SOLAS...

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 cognizant...

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 cognizant...

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 cognizant...

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.

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

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.

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 176.910 Shipping COAST GUARD, DEPARTMENT OF HOMELAND SECURITY (CONTINUED) SMALL PASSENGER VESSELS (UNDER 100 GROSS TONS) INSPECTION AND CERTIFICATION International Convention for Safety of Life at Sea, 1974, as Amended (SOLAS) § 176.910 Passenger Ship...

Passenger well-being in airplanes.

PubMed

Hinninghofen, H; Enck, P

2006-10-30

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

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…

Passenger safety, health, and comfort: a review.

PubMed

Rayman, R B

1997-05-01

Since the birth of aviation medicine approximately 80 yrs ago, practitioners and scientists have given their attention primarily to flight deck crew, cabin crew, and ground support personnel. However, in more recent years we have broadened our horizons to include the safety, health, and comfort of passengers flying commercial aircraft. This will be even more compelling as more passengers take to the air in larger aircraft and flying longer hours to more distant destinations. Further, we can expect to see more older passengers because people in many countries are living longer, healthier lives. The author first discusses the stresses imposed by ordinary commercial flight upon travelers such as airport tumult, barometric pressure changes, immobility, jet lag, noise/ vibration, and radiation. Medical considerations are next addressed describing inflight illness and medical care capability aboard U.S. air carriers. Passenger safety, cabin air quality, and the preventive medicine aspects of air travel are next reviewed in the context of passenger safety, health, and comfort. Recommendations are addressed to regulator agencies, airlines aircraft manufacturers, and the aerospace medicine community.

Identification of influential users by neighbors in online social networks

NASA Astrophysics Data System (ADS)

Sheikhahmadi, Amir; Nematbakhsh, Mohammad Ali; Zareie, Ahmad

2017-11-01

Identification and ranking of influential users in social networks for the sake of news spreading and advertising has recently become an attractive field of research. Given the large number of users in social networks and also the various relations that exist among them, providing an effective method to identify influential users has been gradually considered as an essential factor. In most of the already-provided methods, those users who are located in an appropriate structural position of the network are regarded as influential users. These methods do not usually pay attention to the interactions among users, and also consider those relations as being binary in nature. This paper, therefore, proposes a new method to identify influential users in a social network by considering those interactions that exist among the users. Since users tend to act within the frame of communities, the network is initially divided into different communities. Then the amount of interaction among users is used as a parameter to set the weight of relations existing within the network. Afterward, by determining the neighbors' role for each user, a two-level method is proposed for both detecting users' influence and also ranking them. Simulation and experimental results on twitter data shows that those users who are selected by the proposed method, comparing to other existing ones, are distributed in a more appropriate distance. Moreover, the proposed method outperforms the other ones in terms of both the influential speed and capacity of the users it selects.

46 CFR 168.05-5 - Application of passenger vessel inspection regulations.

Code of Federal Regulations, 2011 CFR

2011-10-01

... applying to passenger vessels in subchapters E (Load Lines), F (Marine Engineering), H (Passenger Vessels), J (Electrical Engineering), K (Small Passenger Vessels Carrying More Than 150 Passengers Or With...

46 CFR 168.05-5 - Application of passenger vessel inspection regulations.

Code of Federal Regulations, 2014 CFR

2014-10-01

... applying to passenger vessels in subchapters E (Load Lines), F (Marine Engineering), H (Passenger Vessels), J (Electrical Engineering), K (Small Passenger Vessels Carrying More Than 150 Passengers Or With...

46 CFR 168.05-5 - Application of passenger vessel inspection regulations.

Code of Federal Regulations, 2010 CFR

2010-10-01

... applying to passenger vessels in subchapters E (Load Lines), F (Marine Engineering), H (Passenger Vessels), J (Electrical Engineering), K (Small Passenger Vessels Carrying More Than 150 Passengers Or With...

46 CFR 168.05-5 - Application of passenger vessel inspection regulations.

Code of Federal Regulations, 2013 CFR

2013-10-01

... applying to passenger vessels in subchapters E (Load Lines), F (Marine Engineering), H (Passenger Vessels), J (Electrical Engineering), K (Small Passenger Vessels Carrying More Than 150 Passengers Or With...

The Drosophila bag of marbles Gene Interacts Genetically with Wolbachia and Shows Female-Specific Effects of Divergence

PubMed Central

Flores, Heather A.; Bubnell, Jaclyn E.; Aquadro, Charles F.; Barbash, Daniel A.

2015-01-01

Many reproductive proteins from diverse taxa evolve rapidly and adaptively. These proteins are typically involved in late stages of reproduction such as sperm development and fertilization, and are more often functional in males than females. Surprisingly, many germline stem cell (GSC) regulatory genes, which are essential for the earliest stages of reproduction, also evolve adaptively in Drosophila. One example is the bag of marbles (bam) gene, which is required for GSC differentiation and germline cyst development in females and for regulating mitotic divisions and entry to spermatocyte differentiation in males. Here we show that the extensive divergence of bam between Drosophila melanogaster and D. simulans affects bam function in females but has no apparent effect in males. We further find that infection with Wolbachia pipientis, an endosymbiotic bacterium that can affect host reproduction through various mechanisms, partially suppresses female sterility caused by bam mutations in D. melanogaster and interacts differentially with bam orthologs from D. melanogaster and D. simulans. We propose that the adaptive evolution of bam has been driven at least in part by the long-term interactions between Drosophila species and Wolbachia. More generally, we suggest that microbial infections of the germline may explain the unexpected pattern of evolution of several GSC regulatory genes. PMID:26291077

Who Are the Most Influential Emergency Physicians on Twitter?

PubMed

Riddell, Jeff; Brown, Alisha; Kovic, Ivor; Jauregui, Joshua

2017-02-01

Twitter has recently gained popularity in emergency medicine (EM). Opinion leaders on Twitter have significant influence on the conversation and content, yet little is known about these opinion leaders. We aimed to describe a methodology to identify the most influential emergency physicians (EP) on Twitter and present a current list. We analyzed 2,234 English-language EPs on Twitter from a previously published list of Twitter accounts generated by a snowball sampling technique. Using NodeXL software, we performed a network analysis of these EPs and ranked them on three measures of influence: in-degree centrality, eigenvector centrality, and betweenness centrality. We analyzed the top 100 users in each of these three measures of influence and compiled a list of users found in the top 100 in all three measures. Of the 300 total users identified by one of the measures of influence, there were 142 unique users. Of the 142 unique users, 61 users were in the top 100 on all three measures of influence. We identify these 61 users as the most influential EM Twitter users. We both describe a method for identifying the most influential users and provide a list of the 61 most influential EPs on Twitter as of January 1, 2016. This application of network science to the EM Twitter community can guide future research to better understand the networked global community of EM.

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

19 CFR 122.156 - Release of passengers.

Code of Federal Regulations, 2010 CFR

2010-04-01

... TREASURY AIR COMMERCE REGULATIONS Flights to and From Cuba § 122.156 Release of passengers. No passengers arriving from Cuba by aircraft will be released by Customs, nor will the aircraft be cleared or permitted...

46 CFR 115.113 - Passengers permitted.

Code of Federal Regulations, 2013 CFR

2013-10-01

..., tables, or other room furnishings; (iii) Toilets and washrooms; (iv) Spaces occupied by and necessary for... 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...

46 CFR 115.113 - Passengers permitted.

Code of Federal Regulations, 2012 CFR

2012-10-01

..., tables, or other room furnishings; (iii) Toilets and washrooms; (iv) Spaces occupied by and necessary for... 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...

46 CFR 115.113 - Passengers permitted.

Code of Federal Regulations, 2014 CFR

2014-10-01

..., tables, or other room furnishings; (iii) Toilets and washrooms; (iv) Spaces occupied by and necessary for... 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...

[Comfort of crew and passengers and atmospheric pressure, noise, wind speed in high-speed train of Shijiazhuang-Taiyuan passenger dedicated line].

PubMed

Zhai, Yi-biao; Huo, Wei; Liu, Qiao-ying; Chen, Bao-shan; Zhang, Jin-long; Shi, Lei

2012-11-01

To explore the crew and passengers' comfort on the Shijiazhuang-Taiyuan passenger dedicated line and physical factors, such as air pressure, noise, wind speed. Comfort investigation of all the crew (n = 244) and passengers (n = 377) on the Shijiazhuang-Taiyuan passenger dedicated line at speed of 250 km/h and 200 km/h and the detection of the air pressure, noise and wind speed were performed in 2011. Significantly higher ratio of comfortable feeling, lower ratio of seriously discomfortable feeling were observed in crew and passengers at 200 km/h compared with those at 250 km/h (P < 0.05), as well as rapid disappearance of discomfortable feeling in crew (P < 0.05) and significantly higher ratio of lightly discomfortable feeling and lower ratios of tinnitus and eardrum discomfort induced by air pressure and noise in passengers at 200 km/h. No significant difference was observed in ear discomfort induced by air pressure and noise among crew, and the duration of disappearance of discomfortable feeling among passengers between 200 km/h and 250 km/h. The noise in carriages exceeded the related standard when the high-speed train passing through the tunnels. The individuals feel more comfortable at 200 km/h than 250 km/h in this line., which may be related with rapid variation of wind speed and noise when the train passes through the tunnels with high speed.

Organizational networks for promoting child passenger safety

DOT National Transportation Integrated Search

1980-10-01

The report describes the organizational network of various support groups that was established by the Tennessee Child Passenger Safety Program as a means of supporting and promoting Tennessee's child passenger protection law. Chapter I introduces the...

Passenger comfort during terminal-area flight maneuvers. M.S. Thesis.

NASA Technical Reports Server (NTRS)

Schoonover, W. E., Jr.

1976-01-01

A series of flight experiments was conducted to obtain passenger subjective responses to closely controlled and repeatable flight maneuvers. In 8 test flights, reactions were obtained from 30 passenger subjects to a wide range of terminal-area maneuvers, including descents, turns, decelerations, and combinations thereof. Analysis of the passenger rating variance indicated that the objective of a repeatable flight passenger environment was achieved. Multiple linear regression models developed from the test data were used to define maneuver motion boundaries for specified degrees of passenger acceptance.

Stress analysis on passenger deck due to modification from passenger ship to vehicle-carrying ship

NASA Astrophysics Data System (ADS)

Zubaydi, A.; Sujiatanti, S. H.; Hariyanto, T. R.

2018-03-01

Stress is a basic concept in learning about material mechanism. The main focus that needs to be brought to attention in analyzing stress is strength, which is the structural capacity to carry or distribute loads. The structural capacity not only measured by comparing the maximum stress with the material’s yield strength but also with the permissible stress required by the Indonesian Classification Bureau (BKI), which certainly makes it much safer. This final project analyzes stress in passenger deck that experiences modification due to load changes, from passenger load to vehicle one, carrying: 6-wheels truck with maximum weight of 14 tons, a passenger car with maximum weight of 3.5 tons, and a motorcycle with maximum weight of 0.4 tons. The deck structure is modelled using finite element software. The boundary conditions given to the structural model are fix and simple constraint. The load that works on this deck is the deck load which comes from the vehicles on deck with three vehicles’ arrangement plans. After that, software modelling is conducted for analysis purpose. Analysis result shows a variation of maximum stress that occurs i.e. 135 N/mm2, 133 N/mm2, and 152 N/mm2. Those maximum stresses will not affect the structure of passenger deck’s because the maximum stress that occurs indicates smaller value compared to the Indonesian Classification Bureau’s permissible stress (175 N/mm2) as well as the material’s yield strength (235 N/mm2). Thus, the structural strength of passenger deck is shown to be capable of carrying the weight of vehicles in accordance with the three vehicles’ arrangement plans.

14 CFR Sec. 19-7 - Passenger origin-destination survey.

Code of Federal Regulations, 2013 CFR

2013-01-01

... 14 Aeronautics and Space 4 2013-01-01 2013-01-01 false Passenger origin-destination survey. Sec... AIR CARRIERS Operating Statistics Classifications Sec. 19-7 Passenger origin-destination survey. (a... carriers) shall participate in a Passenger Origin-Destination (O & D) Survey covering domestic and...

14 CFR Sec. 19-7 - Passenger origin-destination survey.

Code of Federal Regulations, 2014 CFR

2014-01-01

... 14 Aeronautics and Space 4 2014-01-01 2014-01-01 false Passenger origin-destination survey. Sec... AIR CARRIERS Operating Statistics Classifications Sec. 19-7 Passenger origin-destination survey. (a... carriers) shall participate in a Passenger Origin-Destination (O & D) Survey covering domestic and...

Detecting influential observations in nonlinear regression modeling of groundwater flow

USGS Publications Warehouse

Yager, Richard 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.

A Data-Based Approach to Discovering Multi-Topic Influential Leaders.

PubMed

Tang, Xing; Miao, Qiguang; Yu, Shangshang; Quan, Yining

2016-01-01

Recently, increasing numbers of users have adopted microblogging services as their main information source. However, most of them find themselves drowning in the millions of posts produced by other users every day. To cope with this, identifying a set of the most influential people is paramount. Moreover, finding a set of related influential users to expand the coverage of one particular topic is required in real world scenarios. Most of the existing algorithms in this area focus on topology-related methods such as PageRank. These methods mine link structures to find the expected influential rank of users. However, because they ignore the interaction data, these methods turn out to be less effective in social networks. In reality, a variety of topics exist within the information diffusing through the network. Because they have different interests, users play different roles in the diffusion of information related to different topics. As a result, distinguishing influential leaders according to different topics is also worthy of research. In this paper, we propose a multi-topic influence diffusion model (MTID) based on traces acquired from historic information. We decompose the influential scores of users into two parts: the direct influence determined by information propagation along the link structure and indirect influence that extends beyond the restrictions of direct follower relationships. To model the network from a multi-topical viewpoint, we introduce topic pools, each of which represents a particular topic information source. Then, we extract the topic distributions from the traces of tweets, determining the influence propagation probability and content generation probability. In the network, we adopt multiple ground nodes representing topic pools to connect every user through bidirectional links. Based on this multi-topical view of the network, we further introduce the topic-dependent rank (TD-Rank) algorithm to identify the multi-topic influential users

A Data-Based Approach to Discovering Multi-Topic Influential Leaders

PubMed Central

Tang, Xing; Miao, Qiguang; Yu, Shangshang; Quan, Yining

2016-01-01

Recently, increasing numbers of users have adopted microblogging services as their main information source. However, most of them find themselves drowning in the millions of posts produced by other users every day. To cope with this, identifying a set of the most influential people is paramount. Moreover, finding a set of related influential users to expand the coverage of one particular topic is required in real world scenarios. Most of the existing algorithms in this area focus on topology-related methods such as PageRank. These methods mine link structures to find the expected influential rank of users. However, because they ignore the interaction data, these methods turn out to be less effective in social networks. In reality, a variety of topics exist within the information diffusing through the network. Because they have different interests, users play different roles in the diffusion of information related to different topics. As a result, distinguishing influential leaders according to different topics is also worthy of research. In this paper, we propose a multi-topic influence diffusion model (MTID) based on traces acquired from historic information. We decompose the influential scores of users into two parts: the direct influence determined by information propagation along the link structure and indirect influence that extends beyond the restrictions of direct follower relationships. To model the network from a multi-topical viewpoint, we introduce topic pools, each of which represents a particular topic information source. Then, we extract the topic distributions from the traces of tweets, determining the influence propagation probability and content generation probability. In the network, we adopt multiple ground nodes representing topic pools to connect every user through bidirectional links. Based on this multi-topical view of the network, we further introduce the topic-dependent rank (TD-Rank) algorithm to identify the multi-topic influential users

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.

14 CFR 29.1413 - Safety belts: passenger warning device.

Code of Federal Regulations, 2013 CFR

2013-01-01

... 14 Aeronautics and Space 1 2013-01-01 2013-01-01 false Safety belts: passenger warning device. 29.1413 Section 29.1413 Aeronautics and Space FEDERAL AVIATION ADMINISTRATION, DEPARTMENT OF....1413 Safety belts: passenger warning device. (a) If there are means to indicate to the passengers when...

14 CFR 29.1413 - Safety belts: passenger warning device.

Code of Federal Regulations, 2012 CFR

2012-01-01

... 14 Aeronautics and Space 1 2012-01-01 2012-01-01 false Safety belts: passenger warning device. 29.1413 Section 29.1413 Aeronautics and Space FEDERAL AVIATION ADMINISTRATION, DEPARTMENT OF....1413 Safety belts: passenger warning device. (a) If there are means to indicate to the passengers when...

14 CFR 29.1413 - Safety belts: passenger warning device.

Code of Federal Regulations, 2010 CFR

2010-01-01

... 14 Aeronautics and Space 1 2010-01-01 2010-01-01 false Safety belts: passenger warning device. 29.1413 Section 29.1413 Aeronautics and Space FEDERAL AVIATION ADMINISTRATION, DEPARTMENT OF....1413 Safety belts: passenger warning device. (a) If there are means to indicate to the passengers when...

14 CFR 29.1413 - Safety belts: passenger warning device.

Code of Federal Regulations, 2011 CFR

2011-01-01

... 14 Aeronautics and Space 1 2011-01-01 2011-01-01 false Safety belts: passenger warning device. 29.1413 Section 29.1413 Aeronautics and Space FEDERAL AVIATION ADMINISTRATION, DEPARTMENT OF....1413 Safety belts: passenger warning device. (a) If there are means to indicate to the passengers when...

14 CFR 29.1413 - Safety belts: passenger warning device.

Code of Federal Regulations, 2014 CFR

2014-01-01

... 14 Aeronautics and Space 1 2014-01-01 2014-01-01 false Safety belts: passenger warning device. 29.1413 Section 29.1413 Aeronautics and Space FEDERAL AVIATION ADMINISTRATION, DEPARTMENT OF....1413 Safety belts: passenger warning device. (a) If there are means to indicate to the passengers when...

Intercity passenger rail : the congress faces critical decisions about the role of and funding for intercity passenger rail systems

DOT National Transportation Integrated Search

2001-07-25

With the growth in the nation's highway and aviation systems in the : previous decades, intercity passenger rail service lost its competitive edge. Highways have enabled cars to be competitive with conventional passenger trains (those operating up to...

Qualities of Influential Literacy Teacher Educators

ERIC Educational Resources Information Center

Wold, Linda S.; Young, Janet R.; Risko, Victoria J.

2011-01-01

An online survey of award-winning literacy teachers was conducted to determine the most influential qualities of literacy teacher educators in teacher preparation programs. Sixty-two recipients of literacy awards participated in the study, representing teachers of excellence from all U.S. geographic regions. Using a backward mapping process,…

A Veritable Menagerie of Heritable Bacteria from Ants, Butterflies, and Beyond: Broad Molecular Surveys and a Systematic Review

PubMed Central

Russell, Jacob A.; Funaro, Colin F.; Giraldo, Ysabel M.; Goldman-Huertas, Benjamin; Suh, David; Kronauer, Daniel J. C.; Moreau, Corrie S.; Pierce, Naomi E.

2012-01-01

Maternally transmitted bacteria have been important players in the evolution of insects and other arthropods, affecting their nutrition, defense, development, and reproduction. Wolbachia are the best studied among these and typically the most prevalent. While several other bacteria have independently evolved a heritable lifestyle, less is known about their host ranges. Moreover, most groups of insects have not had their heritable microflora systematically surveyed across a broad range of their taxonomic diversity. To help remedy these shortcomings we used diagnostic PCR to screen for five groups of heritable symbionts—Arsenophonus spp., Cardinium hertigii, Hamiltonella defensa, Spiroplasma spp., and Wolbachia spp.—across the ants and lepidopterans (focusing, in the latter case, on two butterfly families—the Lycaenidae and Nymphalidae). We did not detect Cardinium or Hamiltonella in any host. Wolbachia were the most widespread, while Spiroplasma (ants and lepidopterans) and Arsenophonus (ants only) were present at low levels. Co-infections with different Wolbachia strains appeared especially common in ants and less so in lepidopterans. While no additional facultative heritable symbionts were found among ants using universal bacterial primers, microbes related to heritable enteric bacteria were detected in several hosts. In summary, our findings show that Wolbachia are the dominant heritable symbionts of ants and at least some lepidopterans. However, a systematic review of symbiont frequencies across host taxa revealed that this is not always the case across other arthropods. Furthermore, comparisons of symbiont frequencies revealed that the prevalence of Wolbachia and other heritable symbionts varies substantially across lower-level arthropod taxa. We discuss the correlates, potential causes, and implications of these patterns, providing hypotheses on host attributes that may shape the distributions of these influential bacteria. PMID:23284655

Challenging the Wigglesworthia, Sodalis, Wolbachia symbiosis dogma in tsetse flies: Spiroplasma is present in both laboratory and natural populations.

PubMed

Doudoumis, V; Blow, F; Saridaki, A; Augustinos, A; Dyer, N A; Goodhead, I; Solano, P; Rayaisse, J-B; Takac, P; Mekonnen, S; Parker, A G; Abd-Alla, A M M; Darby, A; Bourtzis, K; Tsiamis, G

2017-07-05

Profiling of wild and laboratory tsetse populations using 16S rRNA gene amplicon sequencing allowed us to examine whether the "Wigglesworthia-Sodalis-Wolbachia dogma" operates across species and populations. The most abundant taxa, in wild and laboratory populations, were Wigglesworthia (the primary endosymbiont), Sodalis and Wolbachia as previously characterized. The species richness of the microbiota was greater in wild than laboratory populations. Spiroplasma was identified as a new symbiont exclusively in Glossina fuscipes fuscipes and G. tachinoides, members of the palpalis sub-group, and the infection prevalence in several laboratory and natural populations was surveyed. Multi locus sequencing typing (MLST) analysis identified two strains of tsetse-associated Spiroplasma, present in G. f. fuscipes and G. tachinoides. Spiroplasma density in G. f. fuscipes larva guts was significantly higher than in guts from teneral and 15-day old male and female adults. In gonads of teneral and 15-day old insects, Spiroplasma density was higher in testes than ovaries, and was significantly higher density in live versus prematurely deceased females indicating a potentially mutualistic association. Higher Spiroplasma density in testes than in ovaries was also detected by fluorescent in situ hybridization in G. f. fuscipes.

Who Are the Most Influential Emergency Physicians on Twitter?

PubMed Central

Riddell, Jeff; Brown, Alisha; Kovic, Ivor; Jauregui, Joshua

2017-01-01

Introduction Twitter has recently gained popularity in emergency medicine (EM). Opinion leaders on Twitter have significant influence on the conversation and content, yet little is known about these opinion leaders. We aimed to describe a methodology to identify the most influential emergency physicians (EP) on Twitter and present a current list. Methods We analyzed 2,234 English-language EPs on Twitter from a previously published list of Twitter accounts generated by a snowball sampling technique. Using NodeXL software, we performed a network analysis of these EPs and ranked them on three measures of influence: in-degree centrality, eigenvector centrality, and betweenness centrality. We analyzed the top 100 users in each of these three measures of influence and compiled a list of users found in the top 100 in all three measures. Results Of the 300 total users identified by one of the measures of influence, there were 142 unique users. Of the 142 unique users, 61 users were in the top 100 on all three measures of influence. We identify these 61 users as the most influential EM Twitter users. Conclusion We both describe a method for identifying the most influential users and provide a list of the 61 most influential EPs on Twitter as of January 1, 2016. This application of network science to the EM Twitter community can guide future research to better understand the networked global community of EM. PMID:28210365

Mortality of German travellers on passenger vessels.

PubMed

Oldenburg, Marcus; Herzog, Jan; Püschel, Klaus; Harth, Volker

2016-01-01

In the past two decades, more and more Germans decided to spend their holidays on a passenger vessel. This study examined the frequencies and causes of deaths of German travellers aboard passenger vessels of all flags. The shipboard deaths of all German travellers within the time period from 1998 to 2008 were counted using the German civil central register in Berlin. The available documentation in this register provides information on frequencies, circumstances and causes of deaths on ships. In the above-mentioned period of time, the total cohort of German travellers on cruise ships is estimated to be 5.97 million persons. During the 11-year examination period, 135 shipboard deaths of German passengers [102 males (75.6%) and 33 females (24.4%)] were recorded. Out of these travellers, 110 died on cruise ships. When considering only the passengers on cruise ships (without those on ferries) an average crude mortality rate of 1.8 per 100,000 German passengers was calculated. The crude mortality rate of shipboard death for males and females was 2.5 and 0.8 per 100,000 German passengers with a mean age of 71.2 years [standard deviation (SD) 16.0 years] and 73.3 years (SD 16.0 years), respectively. Significantly, more deceased travellers older than 70 years were observed on traditional cruise ships and resort vessels than on passenger ferries (P = 0.001). The causes of death were documented in 85 cases (63.0%). Out of these documented deaths, 82 (96.5%) cases were regarded to be natural causes (particularly circulatory diseases) and 3 (3.5%) as unnatural causes (twice drowning and once an accidental fall). In spite of the large proportion of unknown causes of death, this study argues for a high significance of internal causes of deaths among German passengers. Thus, ship's doctors-particularly those on traditional cruise ships-should be well experienced in internal and geriatric medicines. © The Author 2016. Published by Oxford University Press on behalf of

Real-time passenger counting by active linear cameras

NASA Astrophysics Data System (ADS)

Khoudour, Louahdi; Duvieubourg, Luc; Deparis, Jean-Pierre

1996-03-01

The companies operating subways are very much concerned with counting the passengers traveling through their transport systems. One of the most widely used systems for counting passengers consists of a mechanical gate equipped with a counter. However, such simple systems are not able to count passengers jumping above the gates. Moreover, passengers carrying large luggage or bags may meet some difficulties when going through such gates. The ideal solution is a contact-free counting system that would bring more comfort of use for the passengers. For these reasons, we propose to use a video processing system instead of these mechanical gates. The optical sensors discussed in this paper offer several advantages including well defined detection areas, fast response time and reliable counting capability. A new technology has been developed and tested, based on linear cameras. Preliminary results show that this system is very efficient when the passengers crossing the optical gate are well separated. In other cases, such as in compact crowd conditions, reasonable accuracy has been demonstrated. These results are illustrated by means of a number of sequences shot in field conditions. It is our belief that more precise measurements could be achieved, in the case of compact crowd, by other algorithms and acquisition techniques of the line images that we are presently developing.

19 CFR 4.80a - Coastwise transportation of passengers.

Code of Federal Regulations, 2010 CFR

2010-04-01

... 19 Customs Duties 1 2010-04-01 2010-04-01 false Coastwise transportation of passengers. 4.80a... transportation of passengers. (a) For the purposes of this section, the following terms will have the meaning set... to the prohibition in this section is the transportation of passengers between ports in Puerto Rico...

19 CFR 4.80a - Coastwise transportation of passengers.

Code of Federal Regulations, 2014 CFR

2014-04-01

... 19 Customs Duties 1 2014-04-01 2014-04-01 false Coastwise transportation of passengers. 4.80a... transportation of passengers. (a) For the purposes of this section, the following terms will have the meaning set... to the prohibition in this section is the transportation of passengers between ports in Puerto Rico...

19 CFR 4.80a - Coastwise transportation of passengers.

Code of Federal Regulations, 2011 CFR

2011-04-01

... 19 Customs Duties 1 2011-04-01 2011-04-01 false Coastwise transportation of passengers. 4.80a... transportation of passengers. (a) For the purposes of this section, the following terms will have the meaning set... to the prohibition in this section is the transportation of passengers between ports in Puerto Rico...

19 CFR 4.80a - Coastwise transportation of passengers.

Code of Federal Regulations, 2013 CFR

2013-04-01

... 19 Customs Duties 1 2013-04-01 2013-04-01 false Coastwise transportation of passengers. 4.80a... transportation of passengers. (a) For the purposes of this section, the following terms will have the meaning set... to the prohibition in this section is the transportation of passengers between ports in Puerto Rico...

19 CFR 4.80a - Coastwise transportation of passengers.

Code of Federal Regulations, 2012 CFR

2012-04-01

... 19 Customs Duties 1 2012-04-01 2012-04-01 false Coastwise transportation of passengers. 4.80a... transportation of passengers. (a) For the purposes of this section, the following terms will have the meaning set... to the prohibition in this section is the transportation of passengers between ports in Puerto Rico...

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 the...

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 the...

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 the...

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 the...

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, of...

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, of...

Passenger Flow Analysis, 1978. Riverside Line, MBTA.

DOT National Transportation Integrated Search

1981-08-01

In order to complete a set of passenger flow estimates for use in a simulation model of a light rail line, a count of passenger movement was made at randomly selected stations in the underground section. Above-ground stations had been studied a year ...

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.

Identifying influential neighbors in animal flocking

PubMed Central

Jiang, Li; Giuggioli, Luca; Escobedo, Ramón; Sire, Clément; Han, Zhangang

2017-01-01

Schools of fish and flocks of birds can move together in synchrony and decide on new directions of movement in a seamless way. This is possible because group members constantly share directional information with their neighbors. Although detecting the directionality of other group members is known to be important to maintain cohesion, it is not clear how many neighbors each individual can simultaneously track and pay attention to, and what the spatial distribution of these influential neighbors is. Here, we address these questions on shoals of Hemigrammus rhodostomus, a species of fish exhibiting strong schooling behavior. We adopt a data-driven analysis technique based on the study of short-term directional correlations to identify which neighbors have the strongest influence over the participation of an individual in a collective U-turn event. We find that fish mainly react to one or two neighbors at a time. Moreover, we find no correlation between the distance rank of a neighbor and its likelihood to be influential. We interpret our results in terms of fish allocating sequential and selective attention to their neighbors. PMID:29161269

46 CFR 15.905 - Uninspected passenger vessels.

Code of Federal Regulations, 2010 CFR

2010-10-01

... 46 Shipping 1 2010-10-01 2010-10-01 false Uninspected passenger vessels. 15.905 Section 15.905 Shipping COAST GUARD, DEPARTMENT OF HOMELAND SECURITY MERCHANT MARINE OFFICERS AND SEAMEN MANNING REQUIREMENTS Equivalents § 15.905 Uninspected passenger vessels. (a) An individual holding a license or MMC...

46 CFR 15.905 - Uninspected passenger vessels.

Code of Federal Regulations, 2011 CFR

2011-10-01

... 46 Shipping 1 2011-10-01 2011-10-01 false Uninspected passenger vessels. 15.905 Section 15.905 Shipping COAST GUARD, DEPARTMENT OF HOMELAND SECURITY MERCHANT MARINE OFFICERS AND SEAMEN MANNING REQUIREMENTS Equivalents § 15.905 Uninspected passenger vessels. (a) An individual holding a license or MMC...

46 CFR 15.905 - Uninspected passenger vessels.

Code of Federal Regulations, 2013 CFR

2013-10-01

... 46 Shipping 1 2013-10-01 2013-10-01 false Uninspected passenger vessels. 15.905 Section 15.905 Shipping COAST GUARD, DEPARTMENT OF HOMELAND SECURITY MERCHANT MARINE OFFICERS AND SEAMEN MANNING REQUIREMENTS Equivalents § 15.905 Uninspected passenger vessels. (a) An individual holding a license or MMC...

46 CFR 15.905 - Uninspected passenger vessels.

Code of Federal Regulations, 2014 CFR

2014-10-01

... 46 Shipping 1 2014-10-01 2014-10-01 false Uninspected passenger vessels. 15.905 Section 15.905 Shipping COAST GUARD, DEPARTMENT OF HOMELAND SECURITY MERCHANT MARINE OFFICERS AND SEAMEN MANNING REQUIREMENTS Equivalents § 15.905 Uninspected passenger vessels. (a) An individual holding a license or MMC...

46 CFR 15.905 - Uninspected passenger vessels.

Code of Federal Regulations, 2012 CFR

2012-10-01

... 46 Shipping 1 2012-10-01 2012-10-01 false Uninspected passenger vessels. 15.905 Section 15.905 Shipping COAST GUARD, DEPARTMENT OF HOMELAND SECURITY MERCHANT MARINE OFFICERS AND SEAMEN MANNING REQUIREMENTS Equivalents § 15.905 Uninspected passenger vessels. (a) An individual holding a license or MMC...

Salmonella outbreak among railway and airline passengers.

PubMed

Hatakka, M

1992-01-01

A widespread outbreak by Salmonella infantis, infecting a total of 226 people, occurred in Finland at the beginning of August 1986. Of those infected, 107 were railway passengers, 91 were airline passengers and 28 were employed in a food processing establishment. The outbreak among the railway passengers was caused by egg sandwiches, the airline passengers were infected by a meal served on board and the catering employees by the breakfast served in the establishment. The outbreak was caused by food prepared in the establishment's kitchen. The employees' breakfasts had probably been contaminated by an employee who was a symptom-free Salmonella infantis carrier, and a number of the employees subsequently became infected, leading to widespread contamination of the food prepared in the establishment. The spread of the outbreak was further influenced by a heatwave at the time and by shortcomings in the cold storage facilities. The kitchen's hygiene supervision and the quality control of its output were reorganized after the outbreak.

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.

75 FR 45562 - Enhancing Airline Passenger Protections

Federal Register 2010, 2011, 2012, 2013, 2014

2010-08-03

... [Docket No. DOT-OST-2010-0140] RIN No. 2105-AD92 Enhancing Airline Passenger Protections AGENCY: Office of... Friday, except Federal Holidays. Fax: (202) 493-2251. Instructions: You must include the agency name and... enhancing airline passenger protections that proposed to improve the air travel environment for [[Page 45563...

Child Passenger Deaths Involving Alcohol-Impaired Drivers

PubMed Central

Quinlan, Kyran; Shults, Ruth A.; Rudd, Rose A.

2017-01-01

BACKGROUND AND OBJECTIVE Approximately 1 in 5 child passenger deaths in the United States involves an alcohol-impaired driver, most commonly the child’s own driver. The objective of this study was to document recent trends and state-specific rates of these deaths. METHODS A descriptive analysis of 2001–2010 Fatality Analysis Reporting System data for child passengers aged <15 years killed in alcohol-impaired driving crashes. Driver impairment was defined as a blood alcohol concentration of ≥0.08 g/dL. RESULTS During 2001–2010, 2344 children <15 years were killed in crashes involving at least 1 alcohol-impaired driver. Of these children, 1515 (65%) were riding with an impaired driver. Annual deaths among children riding with an alcohol-impaired driver decreased by 41% over the decade. Among the 37 states included in the state-level analysis, Texas (272) and California (135) had the most children killed while riding with an impaired driver and South Dakota (0.98) and New Mexico (0.86) had the highest annualized child passenger death rates (per 100 000 children). Most (61%) child passengers of impaired drivers were unrestrained at the time of the crash. One-third of the impaired drivers did not have a valid driver’s license. CONCLUSIONS Alcohol-impaired driving remains a substantial threat to the safety of child passengers in the United States, and typically involves children being driven by impaired drivers. This risk varies meaningfully among states. To make further progress, states and communities could consider increased use of effective interventions and efforts aimed specifically at protecting child passengers from impaired drivers. PMID:24799550

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 COAST GUARD, DEPARTMENT OF HOMELAND SECURITY MERCHANT MARINE OFFICERS AND SEAMEN MANNING REQUIREMENTS Manning Requirements; Inspected Vessels § 15.530 Large passenger vessels. (a) The owner or operator of a U...

46 CFR 15.530 - Large passenger vessels.

Code of Federal Regulations, 2013 CFR

2013-10-01

... 46 Shipping 1 2013-10-01 2013-10-01 false Large passenger vessels. 15.530 Section 15.530 Shipping COAST GUARD, DEPARTMENT OF HOMELAND SECURITY MERCHANT MARINE OFFICERS AND SEAMEN MANNING REQUIREMENTS Manning Requirements; Inspected Vessels § 15.530 Large passenger vessels. (a) The owner or operator of a U...

46 CFR 15.530 - Large passenger vessels.

Code of Federal Regulations, 2012 CFR

2012-10-01

... 46 Shipping 1 2012-10-01 2012-10-01 false Large passenger vessels. 15.530 Section 15.530 Shipping COAST GUARD, DEPARTMENT OF HOMELAND SECURITY MERCHANT MARINE OFFICERS AND SEAMEN MANNING REQUIREMENTS Manning Requirements; Inspected Vessels § 15.530 Large passenger vessels. (a) The owner or operator of a U...

46 CFR 15.530 - Large passenger vessels.

Code of Federal Regulations, 2011 CFR

2011-10-01

... 46 Shipping 1 2011-10-01 2011-10-01 false Large passenger vessels. 15.530 Section 15.530 Shipping COAST GUARD, DEPARTMENT OF HOMELAND SECURITY MERCHANT MARINE OFFICERS AND SEAMEN MANNING REQUIREMENTS Manning Requirements; Inspected Vessels § 15.530 Large passenger vessels. (a) The owner or operator of a U...

46 CFR 15.530 - Large passenger vessels.

Code of Federal Regulations, 2014 CFR

2014-10-01

... 46 Shipping 1 2014-10-01 2014-10-01 false Large passenger vessels. 15.530 Section 15.530 Shipping COAST GUARD, DEPARTMENT OF HOMELAND SECURITY MERCHANT MARINE OFFICERS AND SEAMEN MANNING REQUIREMENTS Manning Requirements; Inspected Vessels § 15.530 Large passenger vessels. (a) The owner or operator of a U...

Reducing risks to children in vehicles with passenger airbags.

PubMed

Graham, J D; Goldie, S J; Segui-Gomez, M; Thompson, K M; Nelson, T; Glass, R; Simpson, A; Woerner, L G

1998-07-01

This review examines the risk that passenger airbags pose for children and discusses behavioral and technologic measures aimed at protecting children from airbag deployment. Although airbags reduce fatal crash injuries among adult drivers and passengers, this safety technology increases mortality risk among children younger than age 12. The magnitude of the risk is multiplied when children are unrestrained or restrained improperly. As new vehicles are resold to buyers who tend to be less safety-conscious than new car owners, the number of children endangered by passenger airbag deployment may increase. For vehicles already in the fleet, strong measures are required to secure children in the rear seat and increase the proper use of appropriate restraint systems through police enforcement of laws. One promising strategy is to amend child passenger safety laws to require that parents secure children in the rear seats. For future vehicles, a mandatory performance standard should be adopted that suppresses airbag deployment automatically if a child is located in the front passenger seat. Other promising improvements in airbag design also are discussed. Major changes in passenger airbag design must be evaluated in a broad analytical framework that considers the welfare of adults as well as children.

Manikin families representing obese airline passengers in the US.

PubMed

Park, Hanjun; Park, Woojin; Kim, Yongkang

2014-01-01

Aircraft passenger spaces designed without proper anthropometric analyses can create serious problems for obese passengers, including: possible denial of boarding, excessive body pressures and contact stresses, postural fixity and related health hazards, and increased risks of emergency evacuation failure. In order to help address the obese passenger's accommodation issues, this study developed male and female manikin families that represent obese US airline passengers. Anthropometric data of obese individuals obtained from the CAESAR anthropometric database were analyzed through PCA-based factor analyses. For each gender, a 99% enclosure cuboid was constructed, and a small set of manikins was defined on the basis of each enclosure cuboid. Digital human models (articulated human figures) representing the manikins were created using a human CAD software program. The manikin families were utilized to develop design recommendations for selected aircraft seat dimensions. The manikin families presented in this study would greatly facilitate anthropometrically accommodating large airline passengers.

Finding the top influential bloggers based on productivity and popularity features

NASA Astrophysics Data System (ADS)

Khan, Hikmat Ullah; Daud, Ali

2017-07-01

A blog acts as a platform of virtual communication to share comments or views about products, events and social issues. Like other social web activities, blogging actions spread to a large number of people. Users influence others in many ways, such as buying a product, having a particular political or social opinion or initiating new activity. Finding the top influential bloggers is an active research domain as it helps us in various fields, such as online marketing, e-commerce, product search and e-advertisements. There exist various models to find the influential bloggers, but they consider limited features using non-modular approach. This paper proposes a new model, Popularity and Productivity Model (PPM), based on a modular approach to find the top influential bloggers. It consists of popularity and productivity modules which exploit various features. We discuss the role of each proposed and existing features and evaluate the proposed model against the standard baseline models using datasets from the real-world blogs. The analysis using standard performance evaluation measures verifies that both productivity and popularity modules play a vital role to find influential bloggers in blogging community in an effective manner.

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…

78 FR 38102 - Passenger Vessels Accessibility Guidelines

Federal Register 2010, 2011, 2012, 2013, 2014

2013-06-25

... translation (CART) web streaming link will be posted on the Access Board's Passenger Vessels homepage at http... guidelines use mandatory language (i.e., shall) so DOT and DOJ can adopt the guidelines as accessibility... passenger vessels subject to U.S. Coast Guard regulations in 46 CFR Chapter I, Subchapters H and K.\\6...

Intercity passenger rail : prospects for Amtrak's financial viability

DOT National Transportation Integrated Search

1998-06-01

Since it began operations in 1971, the National Railroad Passenger Corporation (AmtraK0 has never been profitable and has received about $21 billion in federal subsidies for operating and capital expenses. Amtrak currently provides intercity passenge...

Airbags & children: making correct choices in child passenger restraints.

PubMed

Kamerling, Susan Nudelman

2002-01-01

Countless numbers of young lives are lost each year due to motor vehicle crashes. One of the most effective means of reducing the number of children killed and injured as occupants in motor vehicles is through the use of child passenger restraints. Seat belts and child safety seats save lives and reduce the severity of injuries sustained by occupants in motor vehicle crashes. Nurses are in a unique position to educate and influence families on the proper way to safely transport all motor vehicle passengers. Therefore, nurses working with infants, children, and families have a moral and professional obligation to understand the basics of child passenger safety. At the very least, nurses should be able to direct families appropriately for specifics related to child passenger restraints. The intent of this article is to provide an overview of child passenger safety through a historical, theoretical, and clinical approach. Proper child restraint use is reviewed based on current recommendations for age and size. The potential hazards of child restraint misuse, with an in-depth discussion on airbags, is presented. The discussion on airbags serves to dispel any misconceptions that may be held about airbags. The article includes two case studies that illustrate the injury potential of relatively common misuse patterns. The clinical implications for maternal-child nurses include the recognition that child passenger deaths are primarily due to child restraint nonuse and misuse and the realization of nursing's role in the promotion of proper child passenger restraint.

49 CFR 231.14 - Passenger-train cars without end platforms.

Code of Federal Regulations, 2010 CFR

2010-10-01

... 49 Transportation 4 2010-10-01 2010-10-01 false Passenger-train cars without end platforms. 231.14... ADMINISTRATION, DEPARTMENT OF TRANSPORTATION RAILROAD SAFETY APPLIANCE STANDARDS § 231.14 Passenger-train cars without end platforms. (a) Handbrakes—(1) Number. Each passenger-train car shall be equipped with an...

49 CFR 231.14 - Passenger-train cars without end platforms.

Code of Federal Regulations, 2011 CFR

2011-10-01

... 49 Transportation 4 2011-10-01 2011-10-01 false Passenger-train cars without end platforms. 231.14... ADMINISTRATION, DEPARTMENT OF TRANSPORTATION RAILROAD SAFETY APPLIANCE STANDARDS § 231.14 Passenger-train cars without end platforms. (a) Handbrakes—(1) Number. Each passenger-train car shall be equipped with an...

49 CFR 231.14 - Passenger-train cars without end platforms.

Code of Federal Regulations, 2013 CFR

2013-10-01

... 49 Transportation 4 2013-10-01 2013-10-01 false Passenger-train cars without end platforms. 231.14... ADMINISTRATION, DEPARTMENT OF TRANSPORTATION RAILROAD SAFETY APPLIANCE STANDARDS § 231.14 Passenger-train cars without end platforms. (a) Handbrakes—(1) Number. Each passenger-train car shall be equipped with an...

49 CFR 231.12 - Passenger-train cars with wide vestibules.

Code of Federal Regulations, 2013 CFR

2013-10-01

... 49 Transportation 4 2013-10-01 2013-10-01 false Passenger-train cars with wide vestibules. 231.12... ADMINISTRATION, DEPARTMENT OF TRANSPORTATION RAILROAD SAFETY APPLIANCE STANDARDS § 231.12 Passenger-train cars with wide vestibules. (a) Hand brakes—(1) Number. Each passenger-train car shall be equipped with an...

49 CFR 231.12 - Passenger-train cars with wide vestibules.

Code of Federal Regulations, 2011 CFR

2011-10-01

... 49 Transportation 4 2011-10-01 2011-10-01 false Passenger-train cars with wide vestibules. 231.12... ADMINISTRATION, DEPARTMENT OF TRANSPORTATION RAILROAD SAFETY APPLIANCE STANDARDS § 231.12 Passenger-train cars with wide vestibules. (a) Hand brakes—(1) Number. Each passenger-train car shall be equipped with an...

49 CFR 231.12 - Passenger-train cars with wide vestibules.

Code of Federal Regulations, 2014 CFR

2014-10-01

... 49 Transportation 4 2014-10-01 2014-10-01 false Passenger-train cars with wide vestibules. 231.12... ADMINISTRATION, DEPARTMENT OF TRANSPORTATION RAILROAD SAFETY APPLIANCE STANDARDS § 231.12 Passenger-train cars with wide vestibules. (a) Hand brakes—(1) Number. Each passenger-train car shall be equipped with an...

49 CFR 231.14 - Passenger-train cars without end platforms.

Code of Federal Regulations, 2014 CFR

2014-10-01

... 49 Transportation 4 2014-10-01 2014-10-01 false Passenger-train cars without end platforms. 231.14... ADMINISTRATION, DEPARTMENT OF TRANSPORTATION RAILROAD SAFETY APPLIANCE STANDARDS § 231.14 Passenger-train cars without end platforms. (a) Handbrakes—(1) Number. Each passenger-train car shall be equipped with an...

49 CFR 231.12 - Passenger-train cars with wide vestibules.

Code of Federal Regulations, 2010 CFR

2010-10-01

... 49 Transportation 4 2010-10-01 2010-10-01 false Passenger-train cars with wide vestibules. 231.12... ADMINISTRATION, DEPARTMENT OF TRANSPORTATION RAILROAD SAFETY APPLIANCE STANDARDS § 231.12 Passenger-train cars with wide vestibules. (a) Hand brakes—(1) Number. Each passenger-train car shall be equipped with an...

49 CFR 231.12 - Passenger-train cars with wide vestibules.

Code of Federal Regulations, 2012 CFR

2012-10-01

... 49 Transportation 4 2012-10-01 2012-10-01 false Passenger-train cars with wide vestibules. 231.12... ADMINISTRATION, DEPARTMENT OF TRANSPORTATION RAILROAD SAFETY APPLIANCE STANDARDS § 231.12 Passenger-train cars with wide vestibules. (a) Hand brakes—(1) Number. Each passenger-train car shall be equipped with an...

49 CFR 231.14 - Passenger-train cars without end platforms.

Code of Federal Regulations, 2012 CFR

2012-10-01

... 49 Transportation 4 2012-10-01 2012-10-01 false Passenger-train cars without end platforms. 231.14... ADMINISTRATION, DEPARTMENT OF TRANSPORTATION RAILROAD SAFETY APPLIANCE STANDARDS § 231.14 Passenger-train cars without end platforms. (a) Handbrakes—(1) Number. Each passenger-train car shall be equipped with an...

Effects of aircraft cabin noise on passenger comfort.

PubMed

Pennig, Sibylle; Quehl, Julia; Rolny, Vinzent

2012-01-01

The effects of cabin noise on subjective comfort assessments were systematically investigated in order to reveal optimisation potentials for an improved passenger noise acceptance. Two aircraft simulation studies were conducted. An acoustic laboratory test facility provided with loudspeaker systems for realistic sound presentations and an aircraft cabin simulator (Dornier Do 728) with a high degree of ecological validity were used. Subjects were exposed to nine different noise patterns (three noise levels ranging from 66 to 78 dB(A) combined with three different frequency spectra). Regression analysis demonstrated a significant increase of passengers' acceptance with lower noise levels and significant effects of different frequency spectra determined by seat position in the aircraft cabin (front, middle, rear). Acoustic cabin design should therefore consider measures beyond noise level reduction altering noise characteristics to improve passengers' comfort and well-being in the aircraft cabin. To improve passenger comfort in the aircraft with respect to cabin noise, passengers' reactions to specific noise conditions were systematically investigated. Two laboratory studies showed significant dose-response relationships between sound pressure level and subjective comfort ratings which differed due to the noise at specific seat positions in the aircraft.

Age Appropriate Restraints for the Right Front Passenger

PubMed Central

Augenstein, J; Perdeck, E.; Digges, K.; Bahouth, G.

2007-01-01

This study applies NASS/CDS, GES and FARS data to examine occupant exposure plus injury and fatality rates for belted occupants in frontal crashes by seating position, age and gender. The NASS data was used to examine the distributions by crash severity. The GES data showed that when two elderly occupants (age 65+) were present, the female occupied the right front passenger position 73% of the time. A paired comparison analysis using FARS data showed that, for elderly occupants (age 65+), the fatality risk for elderly right front passengers is 42% higher than for elderly drivers. The NASS/CDS analysis found 74% of the seriously injured vulnerable passengers with MAIS 3+ injuries were in crashes less severe than 26 mph. This group of injured occupants was made up of 43% aged 50 and older and 42% younger females. The injury rates for the older (age 50+) right front passengers were 1.8 times the rates for the elderly drivers. These results suggest the need for more benign safety systems for the right front passenger that are appropriate for the lower injury tolerance of the predominant occupants of that seating position. PMID:18184503

Passenger and cell phone conversations in simulated driving.

PubMed

Drews, Frank A; Pasupathi, Monisha; Strayer, David L

2008-12-01

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 instructed to converse with a friend about past experiences in which their life was threatened. The results show that the number of driving errors was highest in the cell phone condition; in passenger conversations more references were made to traffic, and the production rate of the driver and the complexity of speech of both interlocutors dropped in response to an increase in the demand of the traffic. The results indicate that passenger conversations differ from cell phone conversations because the surrounding traffic not only becomes a topic of the conversation, helping driver and passenger to share situation awareness, but the driving condition also has a direct influence on the complexity of the conversation, thereby mitigating the potential negative effects of a conversation on driving. PsycINFO Database Record (c) 2008 APA, all rights reserved.

Peer passenger influences on male adolescent drivers’ visual scanning behavior during simulated driving

PubMed Central

Pradhan, Anuj K.; Li, Kaigang; Bingham, C. Raymond; Simons-Morton, Bruce; Ouimet, Marie Claude; Shope, Jean T.

2014-01-01

Purpose There is a higher likelihood of crashes and fatalities when an adolescent drives with peer passengers, especially for male drivers and male passengers. Simulated driving of male adolescent drivers with male peer passengers was studied to examine passenger influences on distraction and inattention. Methods Male adolescents drove in a high-fidelity driving simulator with a male confederate who posed either as a risk-accepting or risk-averse passenger. Drivers’ eye-movements were recorded. The visual scanning behavior of the drivers was compared when driving alone versus when driving with a passenger, and when driving with a risk-accepting versus a risk-averse passenger. Results The visual scanning of a driver significantly narrowed horizontally and vertically when driving with a peer passenger. There were no significant differences in the times the drivers’ eyes were off the forward roadway when driving with a passenger versus when driving alone. Some significant correlations were found between personality characteristics and the outcome measures. Conclusions The presence of a male peer passenger was associated with a reduction in the visual scanning range of male adolescent drivers. This reduction could be a result of potential cognitive load imposed on the driver due to the presence of a passenger and the real or perceived normative influences or expectations from the passenger. Implications and contribution The presence of male peer passengers was associated with deficient visual scanning in male adolescent drivers. Such reduced scanning behavior is evident in drivers with high cognitive load. Further investigation of passenger influences on adolescent drivers should include examination of distraction and inattention aspects of passenger influence. PMID:24759440

Spatial and Temporal Variation in Aedes aegypti and Aedes albopictus (Diptera: Culicidae) Numbers in the Yogyakarta Area of Java, Indonesia, With Implications for Wolbachia Releases.

PubMed

Tantowijoyo, W; Arguni, E; Johnson, P; Budiwati, N; Nurhayati, P I; Fitriana, I; Wardana, S; Ardiansyah, H; Turley, A P; Ryan, P; O'Neill, S L; Hoffmann, A A

2016-01-01

of mosquito vector populations, particularly through Wolbachia endosymbionts. The success of these strategies depends on understanding the dynamics of vector populations. In preparation for Wolbachia releases around Yogyakarta, we have studied Aedes populations in five hamlets. Adult monitoring with BioGent- Sentinel (BG-S) traps indicated that hamlet populations had different dynamics across the year; while there was an increase in Aedes aegypti (L.) and Aedes albopictus (Skuse) numbers in the wet season, species abundance remained relatively stable in some hamlets but changed markedly (>2 fold) in others. Local rainfall a month prior to monitoring partly predicted numbers of Ae. aegypti but not Ae. albopictus. Site differences in population size indicated by BG-S traps were also evident in ovitrap data. Egg or larval collections with ovitraps repeated at the same location suggested spatial autocorrelation (<250 m) in the areas of the hamlets where Ae. aegypti numbers were high. Overall, there was a weak negative association (r<0.43) between Ae. aegypti and Ae. albopictus numbers in ovitraps when averaged across collections. Ae. albopictus numbers in ovitraps and BG-S traps were positively correlated with vegetation around areas where traps were placed, while Ae. aegypti were negatively correlated with this feature. These data inform intervention strategies by defining periods when mosquito densities are high, highlighting the importance of local site characteristics on populations, and suggesting relatively weak interactions between Ae. aegypti and Ae. albopictus. They also indicate local areas within hamlets where consistently high mosquito densities may influence Wolbachia invasions and other interventions.

Passenger flows in underground railway stations and platforms.

DOT National Transportation Integrated Search

2015-05-01

Urban rail systems are designed to carry large volumes of people into and out of major activity centers. As a result, the stations : at these major activity centers are often crowded with boarding and alighting passengers, resulting in passenger inco...

Ocean passenger vessels : migrating south for the winter

DOT National Transportation Integrated Search

2010-01-01

In response to consumer demand, the passenger vessels that operate from seaports along the Atlantic, Gulf, and Pacific coasts alternate between north and south. Passenger vessels that sail out of ports such as New York, Baltimore and Seattle in the s...

The impact of a child passenger restraint law and a public information and education program on child passenger safety in Tennessee

DOT National Transportation Integrated Search

1980-10-01

This report provides an analysis of the impact of child passenger protection legislation and a public information and education (PI&E) program on child passenger safety in Tennessee. This study is unique in that Tennessee was the first state to pass ...

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. 23.1524 Section 23.1524 Aeronautics and Space FEDERAL AVIATION ADMINISTRATION, DEPARTMENT OF... Operating Limitations and Information § 23.1524 Maximum passenger seating configuration. The maximum...

Combining the Sterile Insect Technique with the Incompatible Insect Technique: I-Impact of Wolbachia Infection on the Fitness of Triple- and Double-Infected Strains of Aedes albopictus

PubMed Central

Zhang, Dongjing; Zheng, Xiaoying; Xi, Zhiyong; Bourtzis, Kostas; Gilles, Jeremie R. L.

2015-01-01

The mosquito species Aedes albopictus is a major vector of the human diseases dengue and chikungunya. Due to the lack of efficient and sustainable methods to control this mosquito species, there is an increasing interest in developing and applying the sterile insect technique (SIT) and the incompatible insect technique (IIT), separately or in combination, as population suppression approaches. Ae. albopictus is naturally double-infected with two Wolbachia strains, wAlbA and wAlbB. A new triple Wolbachia-infected strain (i.e., a strain infected with wAlbA, wAlbB, and wPip), known as HC and expressing strong cytoplasmic incompatibility (CI) in appropriate matings, was recently developed. In the present study, we compared several fitness traits of three Ae. albopictus strains (triple-infected, double-infected and uninfected), all of which were of the same genetic background (“Guangzhou City, China”) and were reared under the same conditions. Investigation of egg-hatching rate, survival of pupae and adults, sex ratio, duration of larval stages (development time from L1 to pupation), time to emergence (development time from L1 to adult emergence), wing length, female fecundity and adult longevity indicated that the presence of Wolbachia had only a minimal effect on host fitness. Based on this evidence, the HC strain is currently under consideration for mass rearing and application in a combined SIT-IIT strategy to control natural populations of Ae. albopictus in mainland China. PMID:25849812

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…

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.

Novel strain of Spiroplasma found in flower bugs of the genus Orius (Hemiptera: Anthocoridae): transovarial transmission, coexistence with Wolbachia and varied population density.

PubMed

Watanabe, Masaya; Yukuhiro, Fumiko; Maeda, Taro; Miura, Kazuki; Kageyama, Daisuke

2014-01-01

Spiroplasma, a group of small, wall-less, helical, and motile bacteria belonging to the Mollicutes, contains species with diverse life histories. To date, all the Spiroplasma strains that are known to be transmitted vertically in arthropod lineages belong to either the Spiroplasma ixodetis group or the Spiroplasma poulsonii group. Here, we found that a unique strain of Spiroplasma vertically transmitted in predatory flower bugs of the genus Orius belongs to the Spiroplasma insolitum group, which is a group of bacteria phylogenetically closely related to S. insolitum derived from the tickseed sunflower, Bidens sp. (Asterales: Asteraceae). The infection frequencies in natural populations were16.0% in Orius sauteri (n = 75), 40.5% in Orius nagaii (n = 37), and 8.0% in Orius minutus (n = 87). Orius strigicollis was not infected with Spiroplasma (n = 147). In the early stage of oogenesis (i.e., within the germarium), a large number of bacteria with the typical morphology of Spiroplasma existed, keeping a distance from Wolbachia bacteria. The Spiroplasma population seemed to increase during host development but Wolbachia population did not.

Methods for analysis of passenger trip performance in a complex networked transportation system

NASA Astrophysics Data System (ADS)

Wang, Danyi

2007-12-01

The purpose of the Air Transportation System (ATS) is to provide safe and efficient transportation service of passengers and cargo. The on-time performance of a passenger's trip is a critical performance measurement of the Quality of Service (QOS) provided by any Air Transportation System. QOS has been correlated with airline profitability, productivity, customer loyalty and customer satisfaction (Heskett et al. 1994). Btatu and Barnhart have shown that official government and airline on-time performance metrics (i.e. flight-centric measures of air transportation) fail to accurately reflect the passenger experience (Btatu and Barnhart, 2005). Flight-based metrics do not include the trip delays accrued by passengers who were re-booked due to cancelled flights or missed connections. Also, flight-based metrics do not quantify the magnitude of the delay (only the likelihood) and thus fails to provide the consumer with a useful assessment of the impact of a delay. Passenger-centric metrics have not been developed because of the unavailability of airline proprietary data, which is also protected by anti-trust collusion concerns and civil liberty privacy restrictions. Moveover, the growth of the ATS is trending out of the historical range. The objectives of this research were to (1) estimate ATS-wide passenger trip delay using publicly accessible flight data, and (2) investigate passenger trip dynamics out of the range of historical data by building a passenger flow simulation model to predict impact on passenger trip time given anticipated changes in the future. The first objective enables researchers to conduct historical analysis on passenger on-time performance without proprietary itinerary data, and the second objective enables researchers to conduct experiments outside the range of historic data. The estimated passenger trip delay was for 1,030 routes between the 35 busiest airports in the United States in 2006. The major findings of this research are listed as

Diarrhea and related factors among passengers on world cruises departing from Japan.

PubMed

Yamakawa, Michiyo; Sasai, Megumi; Kasai, Yosuke; Tsuda, Toshihide; Suzuki, Etsuji

2018-01-25

Despite growth in the number of cruises worldwide, evidence about diarrhea experienced by cruise ship passengers remains sparse. We investigated rates of diarrhea and related factors among passengers on world cruises departing from Japan. Targeting passengers on five world cruises (n = 4180) from 2012 to 2013 (85-103 travel days), we calculated rates of health seeking behavior for diarrhea by sex, age group, and number of roommates for each cruise. We estimated rate ratios and 95% confidence intervals, using the group aged 20-39 years, women, and 2-4 roommates as referent categories. We found 5.04-6.00 cases per 10,000 person-days in the five cruises, with an elevated number after calling at ports. Older passengers (>60 years) and passengers with fewer roommates had an elevated risk of health seeking behavior for diarrhea, although passengers aged <20 years had an elevated risk on one cruise. After controlling for covariates (including cruise), significant associations remained for passengers aged >60 years and without roommates. Older passengers and passengers with fewer roommates may be more likely to seek medical treatment for diarrhea during travel on a world cruise, and should take preventive measures. Copyright © 2018. Published by Elsevier Ltd.

Cabin location and the likelihood of motion sickness in cruise ship passengers.

PubMed

Gahlinger , P M

2000-01-01

The prevalence of motion sickness approaches 100% on rough seas. Some previous studies have reported a strong association between location on a ship and the risk of motion sickness, whereas other studies found no association. This study was undertaken to determine if there is a statistical association between the location of the passenger cabin on a ship and the risk of motion sickness in unadapted passengers. Data were collected on 260 passengers on an expedition ship traversing the Drake Passage between South America and Antarctica, during rough sea conditions. A standard scale was employed to record motion sickness severity. The risk of motion sickness was found to be statistically associated with age and sex. However, no association was found with the location of the passenger cabin. Previous research reporting a strong association of motion sickness and passenger location on a ship, studied passengers in the seated position. Passengers who are able to lie in a supine position are at considerably reduced risk of motion sickness. Expedition or cruise ships that provide ready access to berths, allow passengers to avoid the most nauseogenic positions. The location of the passenger cabin does not appear to be related to the likelihood of seasickness.

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

Cape Lookout National Seashore passenger ferry transportation feasibility study

DOT National Transportation Integrated Search

2010-09-01

The Cape Lookout National Seashore Passenger Ferry Transportation Feasibility Study is a technical study documenting the capital investment and operational considerations associated with consolidation of passenger ferry service from Harkers Island an...

Ejection of a rear facing, golf cart passenger.

PubMed

Schau, Kyle; Masory, Oren

2013-10-01

The following report details the findings of a series of experiments and simulations performed on a commercially available, shuttle style golf cart during several maneuvers involving rapid accelerations of the vehicle. It is determined that the current set of passive restraints on these types of golf carts are not adequate in preventing ejection of a rear facing passenger during rapid accelerations in the forward and lateral directions. Experimental data and simulations show that a hip restraint must be a minimum of 13 in. above the seat in order to secure a rear facing passenger during sharp turns, compared to the current restraint height of 5 in. Furthermore, it is determined that a restraint directly in front of the rear facing passenger is necessary to prevent ejection. In addressing these issues, golf cart manufacturers could greatly reduce the likelihood of injury due to ejection of a rear facing, golf cart passenger. Copyright © 2013 Elsevier Ltd. All rights reserved.

Symptom-based categorization of in-flight passenger medical incidents.

PubMed

Mahony, Paul H; Myers, Julia A; Larsen, Peter D; Powell, David M C; Griffiths, Robin F

2011-12-01

The majority of in-flight passenger medical events are managed by cabin crew. Our study aimed to evaluate the reliability of cabin crew reports of in-flight medical events and to develop a symptom-based categorization system. All cabin crew in-flight passenger medical incident reports for an airline over a 9-yr period were examined retrospectively. Validation of incident descriptions were undertaken on a sample of 162 cabin crew reports where medically trained persons' reports were available for comparison using a three Round Delphi technique and testing concordance using Cohen's Kappa. A hierarchical symptom-based categorization system was designed and validated. The rate was 159 incidents per 106 passengers carried, or 70.4/113.3 incidents per 106 revenue passenger kilometres/miles, respectively. Concordance between cabin crew and medical reports was 96%, with a high validity rating (mean 4.6 on a 1-5 scale) and high Cohen's Kappa (0.94). The most common in-flight medical events were transient loss of consciousness (41%), nausea/vomiting/diarrhea (19.5%), and breathing difficulty (16%). Cabin crew records provide reliable data regarding in-flight passenger medical incidents, complementary to diagnosis-based systems, and allow the use of currently underutilized data. The categorization system provides a means for tracking passenger medical incidents internationally and an evidence base for cabin crew first aid training.

Comfort studies of rail passengers

PubMed Central

Nicol, J. F.; Doré, C.; Weiner, J. S.; Lee, D. E.; Prestidge, S. P.; Andrews, M. J.

1973-01-01

Nicol, J. F., Doré, C., Weiner, J. S., Lee, D. E., Prestidge, S. P., and Andrews, M. J. (1973).British Journal of Industrial Medicine,30, 325-334. Comfort studies of rail passengers. A short series of trials is described in which a specimen car of the new High Density Rolling Stock was laden with passengers at different densities and under different environmental constraints, designed to simulate `shut-down' conditions. The results suggest that the limit for comfort, 21·8°C corrected effective temperature (CET), proposed by Bell and Watts (1971) is reasonable but that temperatures some 3 or 4°C higher can be tolerated without undue discomfort. The physiological limit for safety recommended by Bell and Watts is a CET of 30·6°C. This will be reached in less than 20 minutes if there is a power failure in warm conditions in crowded trains. An undesirable, possibly dangerous, level of discomfort will be experienced by passengers in ventilated but crowded trains after 30 minutes. In any case it is recommended that the globe temperature in a carriage should not exceed 30°C. Images PMID:4753715

100 Most Influential Publications in Scoliosis Surgery.

PubMed

Zhou, James Jun; Koltz, Michael T; Agarwal, Nitin; Tempel, Zachary J; Kanter, Adam S; Okonkwo, David O; Hamilton, D Kojo

2017-03-01

Bibliometric analysis. To apply the established technique of citation analysis to identify the 100 most influential articles in scoliosis surgery research published between 1900 and 2015. Previous studies have applied the technique of citation analysis to other areas of study. This is the first article to apply this technique to the field of scoliosis surgery. A two-step search of the Thomson Reuters Web of Science was conducted to identify all articles relevant to the field of scoliosis surgery. The top 100 articles with the most citations were identified based on analysis of titles and abstracts. Further statistical analysis was conducted to determine whether measures of author reputation and overall publication influence affected the rate at which publications were recognized and incorporated by other researchers in the field. Total citations for the final 100 publications included in the list ranged from 82 to 509. The period for publication ranged from 1954 to 2010. Most studies were published in the journal Spine (n = 63). The most frequently published topics of study were surgical techniques (n = 35) and outcomes (n = 35). Measures of author reputation (number of total studies in the top 100, number of first-author studies in the top 100) were found to have no effect on the rate at which studies were adopted by other researchers (number of years until first citation, and number of years until maximum citations). The number of citations/year a publication received was found to be negatively correlated with the rate at which it was adopted by other researchers, indicating that more influential manuscripts attained more rapid recognition by the scientific community at large. In assembling this publication, we have strived to identify and recognize the 100 most influential articles in scoliosis surgery research from 1900 to 2015. N/A.

Application of alternative criteria and procedures to passenger railcars.

DOT National Transportation Integrated Search

2015-05-01

Occupied volume integrity (OVI) refers to a : passenger railcars ability to preserve space for : passengers and crew during an accident. The : Federal Railroad Administrations (FRA) Office : of Research and Development (R&D) is : sponsoring res...

The Research of China's Civil Aviation Passenger Multi-Channel Service Technology Platform

NASA Astrophysics Data System (ADS)

Zhibing, Xue; Xinming, Wang

IATA is promoting Simplifying the Business. The traditional passenger services and business process, such as ticketing, airport counters, had a great influence. The airlines have the passenger service and convenience as the next product development requirements. With civil aviation industry and their company's products construction, the authors propose a solution of passenger multi-channel service product platform. The solution is to streamline the business as the breakthrough point, around the convenience of passengers travel services to travelers as the center, using the current mainstream and the latest IT technology to establish passenger service product platform. The solution will promote DCS e-ticketing business development and service channel diversity. In this paper, the research results have been applied in the product platform construction of the authors' company. The practice shows that through traditional business with the latest IT technologies, traditional passenger services into the emerging service model, passenger service product platform has strong advantages and characteristics. Based on the platform, various types of service products is growing rapidly.

Unsupervised Scalable Statistical Method for Identifying Influential Users in Online Social Networks.

PubMed

Azcorra, A; Chiroque, L F; Cuevas, R; Fernández Anta, A; Laniado, H; Lillo, R E; Romo, J; Sguera, C

2018-05-03

Billions of users interact intensively every day via Online Social Networks (OSNs) such as Facebook, Twitter, or Google+. This makes OSNs an invaluable source of information, and channel of actuation, for sectors like advertising, marketing, or politics. To get the most of OSNs, analysts need to identify influential users that can be leveraged for promoting products, distributing messages, or improving the image of companies. In this report we propose a new unsupervised method, Massive Unsupervised Outlier Detection (MUOD), based on outliers detection, for providing support in the identification of influential users. MUOD is scalable, and can hence be used in large OSNs. Moreover, it labels the outliers as of shape, magnitude, or amplitude, depending of their features. This allows classifying the outlier users in multiple different classes, which are likely to include different types of influential users. Applying MUOD to a subset of roughly 400 million Google+ users, it has allowed identifying and discriminating automatically sets of outlier users, which present features associated to different definitions of influential users, like capacity to attract engagement, capacity to attract a large number of followers, or high infection capacity.

Convenient Airports: Point of View of the Passengers

NASA Technical Reports Server (NTRS)

Magri, Adival Aparecido, Jr.; Alves, Claudio Jorge Pinto

2003-01-01

The competition among airlines or among airports aiming at to increase the demand for its services has been more and more incited. Knowledge the perception of the users for the offered services means to meet the customer's needs and expectations in order either to keep the customer, and therefore keep a significant advantage over competitors. The passenger of the air transportation wants rapidity, security and convenience. Convenience can be translated by comfort that the passenger wants for the price that he can pay. In this paper had been identified. as a result of a survey achieved in six Brazilian airports during 2002, the best indicators in the passenger's perception. These indicators among any others were listed m the handbook of Airports Council International (ACI). Distinctive perceptions were observed among passengers with different travel motivations. This survey had been carried through in the airports of Brasilia, Porto Alegre, Salvador. Fortaleza, Curitiba and Bel6m. Considering this survey we can identified the most attractive airport among them. This work is a way to help improve quality of service, in particular, m these six airports of the Brazilian network. The results should be published and made available to all the parties concerned (airport authority, airlines and service providers) and should lead to corrective action when the passenger is not satisfied with the service.

Airline policy for passengers requiring supplemental in-flight oxygen.

PubMed

Walker, Jacqueline; Kelly, Paul T; Beckert, Lutz

2009-05-01

The aim of this study was to investigate the current Australian/New Zealand airline policy on supplemental in-flight oxygen for passengers with lung disease. Fifty-four commercial airlines servicing international routes were surveyed. Information was gathered from airline call centres and web sites. The survey documented individual airline policy on in-flight oxygen delivery, approval schemes, equipment and cost. Of the 54 airlines contacted, 43 (81%) were able to support passengers requiring in-flight oxygen. The majority (88%) of airlines provided a cylinder for passengers to use. Airline policy for calculating the cost of in-flight oxygen differed considerably between carriers. Six (14%) airlines supplied oxygen to passengers free of charge; however, three of these airlines charged for an extra seat. Fifteen airlines (35%) charged on the basis of oxygen supplied, that is, per cylinder. Fourteen airlines (33%) had a flat rate charge per sector. This study confirmed that most airlines can accommodate passengers requiring supplemental oxygen. However, the findings highlight inconsistencies in airline policies and substantial cost differences for supplemental in-flight oxygen. We advocate an industry standardization of policy and cost of in-flight oxygen.

Fuzzy Temporal Logic Based Railway Passenger Flow Forecast Model

PubMed Central

Dou, Fei; Jia, Limin; Wang, Li; Xu, Jie; Huang, Yakun

2014-01-01

Passenger flow forecast is of essential importance to the organization of railway transportation and is one of the most important basics for the decision-making on transportation pattern and train operation planning. Passenger flow of high-speed railway features the quasi-periodic variations in a short time and complex nonlinear fluctuation because of existence of many influencing factors. In this study, a fuzzy temporal logic based passenger flow forecast model (FTLPFFM) is presented based on fuzzy logic relationship recognition techniques that predicts the short-term passenger flow for high-speed railway, and the forecast accuracy is also significantly improved. An applied case that uses the real-world data illustrates the precision and accuracy of FTLPFFM. For this applied case, the proposed model performs better than the k-nearest neighbor (KNN) and autoregressive integrated moving average (ARIMA) models. PMID:25431586

RCA: A route city attraction model for air passengers

NASA Astrophysics Data System (ADS)

Huang, Feihu; Xiong, Xi; Peng, Jian; Guo, Bing; Tong, Bo

2018-02-01

Human movement pattern is a research hotspot of social computing and has practical values in various fields, such as traffic planning. Previous studies mainly focus on the travel activities of human beings on the ground rather than those in the air. In this paper, we use the reservation records of air passengers to explore air passengers' movement characteristics. After analyzing the effect of the route-trip length on the throughput, we find that most passengers eventually return to their original departure city and that the mobility of air passengers is not related to the route length. Based on these characteristics, we present a route city attraction (RCA) model, in which GDP or population is considered for the calculation of the attraction. The sub models of our RCA model show the better prediction performance of throughput than the radiation model and the gravity model.

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…

Identifying influential nodes in complex networks: A node information dimension approach

NASA Astrophysics Data System (ADS)

Bian, Tian; Deng, Yong

2018-04-01

In the field of complex networks, how to identify influential nodes is a significant issue in analyzing the structure of a network. In the existing method proposed to identify influential nodes based on the local dimension, the global structure information in complex networks is not taken into consideration. In this paper, a node information dimension is proposed by synthesizing the local dimensions at different topological distance scales. A case study of the Netscience network is used to illustrate the efficiency and practicability of the proposed method.

Bus-stop Based Real Time Passenger Information System - Case Study Maribor

NASA Astrophysics Data System (ADS)

Čelan, Marko; Klemenčič, Mitja; Mrgole, Anamarija L.; Lep, Marjan

2017-10-01

Real time passenger information system is one of the key element of promoting public transport. For the successful implementation of real time passenger information systems, various components should be considered, such as: passenger needs and requirements, stakeholder involvement, technological solution for tracking, data transfer, etc. This article carrying out designing and evaluation of real time passenger information (RTPI) in the city of Maribor. The design phase included development of methodology for selection of appropriate macro and micro location of the real-time panel, development of a real-time passenger algorithm, definition of a technical specification, financial issues and time frame. The evaluation shows that different people have different requirements; therefore, the system should be adaptable to be used by various types of people, according to the age, the purpose of journey, experience of using public transport, etc. The average difference between perceived waiting time for a bus is 35% higher than the actual waiting time and grow with the headway increase. Experiences from Maribor have shown that the reliability of real time passenger system (from technical point of view) must be close to 100%, otherwise the system may have negative impact on passengers and may discourage the use of public transport. Among considered events of arrivals during the test period, 92% of all prediction were accurate. The cost benefit analysis has focused only on potential benefits from reduced perceived users waiting time and foreseen costs of real time information system in Maribor for 10 years’ period. Analysis shows that the optimal number for implementing real time passenger information system at the bus stops in Maribor is set on 83 bus stops (approx. 20 %) with the highest number of passenger. If we consider all entries at the chosen bus stops, the total perceived waiting time on yearly level could be decreased by about 60,000 hours.

Assessing the Impact of Influential Observations on Multiple Regression Analysis on Human Resource Research.

ERIC Educational Resources Information Center

Bates, Reid A.; Holton, Elwood F., III; Burnett, Michael F.

1999-01-01

A case study of learning transfer demonstrates the possible effect of influential observation on linear regression analysis. A diagnostic method that tests for violation of assumptions, multicollinearity, and individual and multiple influential observations helps determine which observation to delete to eliminate bias. (SK)

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

49 CFR 231.13 - Passenger-train cars with open-end platforms.

Code of Federal Regulations, 2010 CFR

2010-10-01

... 49 Transportation 4 2010-10-01 2010-10-01 false Passenger-train cars with open-end platforms. 231... Passenger-train cars with open-end platforms. (a) Hand brakes—(1) Number. Each passenger-train car shall be...) Location. Each hand brake shall be so located that it can be safely operated while car is in motion. (b...

49 CFR 231.13 - Passenger-train cars with open-end platforms.

Code of Federal Regulations, 2012 CFR

2012-10-01

... 49 Transportation 4 2012-10-01 2012-10-01 false Passenger-train cars with open-end platforms. 231... Passenger-train cars with open-end platforms. (a) Hand brakes—(1) Number. Each passenger-train car shall be...) Location. Each hand brake shall be so located that it can be safely operated while car is in motion. (b...

49 CFR 231.13 - Passenger-train cars with open-end platforms.

Code of Federal Regulations, 2011 CFR

2011-10-01

... 49 Transportation 4 2011-10-01 2011-10-01 false Passenger-train cars with open-end platforms. 231... Passenger-train cars with open-end platforms. (a) Hand brakes—(1) Number. Each passenger-train car shall be...) Location. Each hand brake shall be so located that it can be safely operated while car is in motion. (b...

49 CFR 231.13 - Passenger-train cars with open-end platforms.

Code of Federal Regulations, 2013 CFR

2013-10-01

... 49 Transportation 4 2013-10-01 2013-10-01 false Passenger-train cars with open-end platforms. 231... Passenger-train cars with open-end platforms. (a) Hand brakes—(1) Number. Each passenger-train car shall be...) Location. Each hand brake shall be so located that it can be safely operated while car is in motion. (b...

49 CFR 231.13 - Passenger-train cars with open-end platforms.

Code of Federal Regulations, 2014 CFR

2014-10-01

... 49 Transportation 4 2014-10-01 2014-10-01 false Passenger-train cars with open-end platforms. 231... Passenger-train cars with open-end platforms. (a) Hand brakes—(1) Number. Each passenger-train car shall be...) Location. Each hand brake shall be so located that it can be safely operated while car is in motion. (b...

MIIB: A Metric to Identify Top Influential Bloggers in a Community.

PubMed

Khan, Hikmat Ullah; Daud, Ali; Malik, Tahir Afzal

2015-01-01

Social networking has revolutionized the use of conventional web and has converted World Wide Web into the social web as users can generate their own content. This change has been possible due to social web platforms like forums, wikis, and blogs. Blogs are more commonly being used as a form of virtual communication to express an opinion about an event, product or experience and can reach a large audience. Users can influence others to buy a product, have certain political or social views, etc. Therefore, identifying the most influential bloggers has become very significant as this can help us in the fields of commerce, advertisement and product knowledge searching. Existing approaches consider some basic features, but lack to consider some other features like the importance of the blog on which the post has been created. This paper presents a new metric, MIIB (Metric for Identification of Influential Bloggers), based on various features of bloggers' productivity and popularity. Productivity refers to bloggers' blogging activity and popularity measures bloggers' influence in the blogging community. The novel module of BlogRank depicts the importance of blog sites where bloggers create their posts. The MIIB has been evaluated against the standard model and existing metrics for finding the influential bloggers using dataset from the real-world blogosphere. The obtained results confirm that the MIIB is able to find the most influential bloggers in a more effective manner.

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

Differential risk of injury in child occupants by passenger car classification.

PubMed

Kallan, Michael J; Durbin, Dennis R; Elliott, Michael R; Menon, Rajiv A; Winston, Flaura K

2003-01-01

In the United States, passenger cars are the most common passenger vehicle, yet they vary widely in size and crashworthiness. Using data collected from a population-based sample of crashes in State Farm-insured vehicles, we quantified the risk of injury to child occupants by passenger car size and classification. Injury risk is predicted by vehicle weight; however, there is an increased risk in both Large vs. Luxury and Sports vs. Small cars, despite similar average vehicle weights in both comparisons. Parents who are purchasing passenger cars should strongly consider the size of the vehicle and its crashworthiness.

Differential Risk of Injury in Child Occupants by Passenger Car Classification

PubMed Central

Kallan, Michael J.; Durbin, Dennis R.; Elliott, Michael R.; Menon, Rajiv A.; Winston, Flaura K.

2003-01-01

In the United States, passenger cars are the most common passenger vehicle, yet they vary widely in size and crashworthiness. Using data collected from a population-based sample of crashes in State Farm-insured vehicles, we quantified the risk of injury to child occupants by passenger car size and classification. Injury risk is predicted by vehicle weight; however, there is an increased risk in both Large vs. Luxury and Sports vs. Small cars, despite similar average vehicle weights in both comparisons. Parents who are purchasing passenger cars should strongly consider the size of the vehicle and its crashworthiness. PMID:12941234

Passenger flow analysis of Beijing urban rail transit network using fractal approach

NASA Astrophysics Data System (ADS)

Li, Xiaohong; Chen, Peiwen; Chen, Feng; Wang, Zijia

2018-04-01

To quantify the spatiotemporal distribution of passenger flow and the characteristics of an urban rail transit network, we introduce four radius fractal dimensions and two branch fractal dimensions by combining a fractal approach with passenger flow assignment model. These fractal dimensions can numerically describe the complexity of passenger flow in the urban rail transit network and its change characteristics. Based on it, we establish a fractal quantification method to measure the fractal characteristics of passenger follow in the rail transit network. Finally, we validate the reasonability of our proposed method by using the actual data of Beijing subway network. It has been shown that our proposed method can effectively measure the scale-free range of the urban rail transit network, network development and the fractal characteristics of time-varying passenger flow, which further provides a reference for network planning and analysis of passenger flow.

Child passengers and driver culpability in fatal crashes by driver gender.

PubMed

Maasalo, Ida; Lehtonen, Esko; Pekkanen, Jami; Summala, Heikki

2016-07-03

Studies based on accident statistics generally suggest that the presence of a passenger reduces adult drivers' accident risk. However, passengers have been reported to be a source of distraction in a remarkable portion of distraction-related crashes. Although the effect of passengers on driving performance has been studied extensively, few studies have focused on how a child passenger affects the driver.  A child in a car is a potential distractor for parents, especially for mothers of small children, who often suffer from sleep deficit. The aim of this study was to examine how the presence of child passengers of different ages is associated with a higher driver culpability, which was expected due to child-related distraction and fatigue. The analysis was based on the comprehensive data of fatal crashes studied in-depth by multidisciplinary road accident investigation teams in Finland during 1988-2012. Teams determine the primary party who had the most crucial effect on the origin of the event. We define the primary party as culpable and the others involved as nonculpable drivers. The culpability rate was defined as the percentage of culpable drivers and rates were compared for drivers with a child/teen passenger aged 0-17 years (N = 348), with an adult passenger without children (N = 324), and when driving alone (N = 579), grouped by child age and driver gender.  Drivers with specific risk-related behavior (substantial speeding, driving when intoxicated, unbelted, or without a license) were excluded from the analyses, in order to make the drivers with and without children comparable. Only drivers 26-47 years old were included, representing parents with children 0-9 years of age. Male drivers were less often culpable with 0- to 17-year-old passengers in the car than alone or with adults. This was not the case with female drivers. The gender difference in culpability was most marked with small children age 0-4 years. Female drivers' culpability rate with a 0

49 CFR 223.17 - Identification of equipped locomotives, passenger cars and cabooses.

Code of Federal Regulations, 2013 CFR

2013-10-01

... cars and cabooses. 223.17 Section 223.17 Transportation Other Regulations Relating to Transportation...-LOCOMOTIVES, PASSENGER CARS AND CABOOSES Specific Requirements § 223.17 Identification of equipped locomotives, passenger cars and cabooses. Each locomotive, passenger car and caboose that is fully equipped with glazing...

49 CFR 238.305 - Interior calendar day mechanical inspection of passenger cars.

Code of Federal Regulations, 2014 CFR

2014-10-01

... passenger cars. 238.305 Section 238.305 Transportation Other Regulations Relating to Transportation... Interior calendar day mechanical inspection of passenger cars. (a) Except as provided in paragraph (e) of this section, each passenger car shall receive an interior mechanical inspection at least once each...

49 CFR 223.17 - Identification of equipped locomotives, passenger cars and cabooses.

Code of Federal Regulations, 2014 CFR

2014-10-01

... cars and cabooses. 223.17 Section 223.17 Transportation Other Regulations Relating to Transportation...-LOCOMOTIVES, PASSENGER CARS AND CABOOSES Specific Requirements § 223.17 Identification of equipped locomotives, passenger cars and cabooses. Each locomotive, passenger car and caboose that is fully equipped with glazing...

49 CFR 238.305 - Interior calendar day mechanical inspection of passenger cars.

Code of Federal Regulations, 2010 CFR

2010-10-01

... passenger cars. 238.305 Section 238.305 Transportation Other Regulations Relating to Transportation... Interior calendar day mechanical inspection of passenger cars. (a) Except as provided in paragraph (d) of this section, each passenger car shall receive an interior mechanical inspection at least once each...

49 CFR 223.17 - Identification of equipped locomotives, passenger cars and cabooses.

Code of Federal Regulations, 2012 CFR

2012-10-01

... cars and cabooses. 223.17 Section 223.17 Transportation Other Regulations Relating to Transportation...-LOCOMOTIVES, PASSENGER CARS AND CABOOSES Specific Requirements § 223.17 Identification of equipped locomotives, passenger cars and cabooses. Each locomotive, passenger car and caboose that is fully equipped with glazing...

49 CFR 223.17 - Identification of equipped locomotives, passenger cars and cabooses.

Code of Federal Regulations, 2010 CFR

2010-10-01

... cars and cabooses. 223.17 Section 223.17 Transportation Other Regulations Relating to Transportation...-LOCOMOTIVES, PASSENGER CARS AND CABOOSES Specific Requirements § 223.17 Identification of equipped locomotives, passenger cars and cabooses. Each locomotive, passenger car and caboose that is fully equipped with glazing...

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

Combining the Sterile Insect Technique with Wolbachia-Based Approaches: II- A Safer Approach to Aedes albopictus Population Suppression Programmes, Designed to Minimize the Consequences of Inadvertent Female Release

PubMed Central

Zhang, Dongjing; Lees, Rosemary Susan; Xi, Zhiyong; Gilles, Jeremie R. L.; Bourtzis, Kostas

2015-01-01

Due to the absence of a perfect method for mosquito sex separation, the combination of the sterile insect technique and the incompatible insect technique is now being considered as a potentially effective method to control Aedes albopictus. In this present study first we examine the minimum pupal irradiation dose required to induce complete sterility in Wolbachia triple-infected (HC), double-infected (GUA) and uninfected (GT) female Ae. albopictus. The HC line is a candidate for Ae. albopictus population suppression programmes, but due to the risk of population replacement which characterizes this triple infected line, the individuals to be released need to be additionally irradiated. After determining the minimum irradiation dose required for complete female sterility, we test whether sterilization is sufficient to prevent invasion of the triple infection from the HC females into double-infected (GUA) populations. Our results indicate that irradiated Ae. albopictus HC, GUA and GT strain females have decreased fecundity and egg hatch rate when irradiated, inversely proportional to the dose, and the complete sterilization of females can be acquired by pupal irradiation with doses above 28 Gy. PCR-based analysis of F1 and F2 progeny indicate that the irradiated HC females, cannot spread the new Wolbachia wPip strain into a small cage GUA population, released at a 1:5 ratio. Considering the above results, we conclude that irradiation can be used to reduce the risk of population replacement caused by an unintentional release of Wolbachia triple-infected Ae. albopictus HC strain females during male release for population suppression. PMID:26252474

Passenger baggage object database (PBOD)

NASA Astrophysics Data System (ADS)

Gittinger, Jaxon M.; Suknot, April N.; Jimenez, Edward S.; Spaulding, Terry W.; Wenrich, Steve A.

2018-04-01

Detection of anomalies of interest in x-ray images is an ever-evolving problem that requires the rapid development of automatic detection algorithms. Automatic detection algorithms are developed using machine learning techniques, which would require developers to obtain the x-ray machine that was used to create the images being trained on, and compile all associated metadata for those images by hand. The Passenger Baggage Object Database (PBOD) and data acquisition application were designed and developed for acquiring and persisting 2-D and 3-D x-ray image data and associated metadata. PBOD was specifically created to capture simulated airline passenger "stream of commerce" luggage data, but could be applied to other areas of x-ray imaging to utilize machine-learning methods.

Metro passenger behaviors and their relations to metro incident involvement.

PubMed

Wan, Xin; Li, Qiming; Yuan, Jingfeng; Schonfeld, Paul M

2015-09-01

The frequent incidents caused by metro passengers in China suggest that it is necessary to explore the classification and effects of passenger behaviors and their relations to incident involvement. A metro passenger behavior questionnaire (MPBQ) and a metro station staff questionnaire (MSSQ), both comprising 32 behavior items, were developed and surveyed on a sample of metro passengers (N=579) and metro staff (N=99). Using the MPBQ, the self-reported frequency of each aberrant behavior was measured and subjected to explanatory factor analysis, which revealed a three-factor solution on the 28 retained behavior items: transgressions, self-willed inattentions and abrupt violations. ANOVA was used to examine the effects of demographic and riding profile variables on different types of behaviors. The MSSQ was used to collect metro staff opinions on behavior frequency, severity and entities that might be affected, given that a specific behavior occurred. An importance hierarchy was established over the 32 identified behaviors to determine the most important riding behaviors. Finally, logistic regression showed that riding time, number of stops experienced by a passenger and, more importantly, transgressions and abrupt violations, were significant predictors of incident involvement. The possible explanations and implications of the findings might help in understanding passenger behaviors and targeting metro safety interventions in ways that promote safer operations. Copyright © 2015 Elsevier Ltd. All rights reserved.

Passenger cab car grade crossing impact test report : rail passenger equipment impact tests

DOT National Transportation Integrated Search

2007-10-01

Two full-scale oblique grade crossing impact tests were conducted in June 2002 to compare the crashworthiness performance of alternative corner post designs on rail passenger cab cars. On June 4, 2002, a cab car fitted with an end frame built to pre-...

36 CFR 5.4 - Commercial passenger-carrying motor vehicles.

Code of Federal Regulations, 2011 CFR

2011-07-01

... motor vehicles. 5.4 Section 5.4 Parks, Forests, and Public Property NATIONAL PARK SERVICE, DEPARTMENT OF THE INTERIOR COMMERCIAL AND PRIVATE OPERATIONS § 5.4 Commercial passenger-carrying motor vehicles. (a) The commercial transportation of passengers by motor vehicles except as authorized under a contract or...

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... passengers: Extended overwater operations. (a) In addition to the oral briefing required by § 121.571(a), each certificate holder operating an airplane in extended overwater operations shall ensure that all...

14 CFR 121.573 - Briefing passengers: Extended overwater operations.

Code of Federal Regulations, 2014 CFR

2014-01-01

... 14 Aeronautics and Space 3 2014-01-01 2014-01-01 false Briefing passengers: Extended overwater... passengers: Extended overwater operations. (a) In addition to the oral briefing required by § 121.571(a), each certificate holder operating an airplane in extended overwater operations shall ensure that all...

14 CFR 121.573 - Briefing passengers: Extended overwater operations.

Code of Federal Regulations, 2012 CFR

2012-01-01

... 14 Aeronautics and Space 3 2012-01-01 2012-01-01 false Briefing passengers: Extended overwater... passengers: Extended overwater operations. (a) In addition to the oral briefing required by § 121.571(a), each certificate holder operating an airplane in extended overwater operations shall ensure that all...

14 CFR 121.573 - Briefing passengers: Extended overwater operations.

Code of Federal Regulations, 2013 CFR

2013-01-01

... 14 Aeronautics and Space 3 2013-01-01 2013-01-01 false Briefing passengers: Extended overwater... passengers: Extended overwater operations. (a) In addition to the oral briefing required by § 121.571(a), each certificate holder operating an airplane in extended overwater operations shall ensure that all...

14 CFR 121.573 - Briefing passengers: Extended overwater operations.

Code of Federal Regulations, 2011 CFR

2011-01-01

... 14 Aeronautics and Space 3 2011-01-01 2011-01-01 false Briefing passengers: Extended overwater... passengers: Extended overwater operations. (a) In addition to the oral briefing required by § 121.571(a), each certificate holder operating an airplane in extended overwater operations shall ensure that all...

49 CFR 37.205 - Additional passengers who use wheelchairs.

Code of Federal Regulations, 2010 CFR

2010-10-01

... 49 Transportation 1 2010-10-01 2010-10-01 false Additional passengers who use wheelchairs. 37.205 Section 37.205 Transportation Office of the Secretary of Transportation TRANSPORTATION SERVICES FOR INDIVIDUALS WITH DISABILITIES (ADA) Over-the-Road Buses (OTRBs) § 37.205 Additional passengers who use...

Design of limited-stop service based on the degree of unbalance of passenger demand

PubMed Central

2018-01-01

This paper presents a limited-stop service for a bus fleet to meet the unbalanced demand of passengers on a bus route and to improve the transit service of the bus route. This strategy includes two parts: a degree assessment of unbalanced passenger demand and an optimization of the limited-stop service. The degree assessment of unbalanced passenger demand, which is based on the different passenger demand between stations and the unbalance of passengers within the station, is used to judge whether implementing the limited-stop service is necessary for a bus route. The optimization of limited-stop service considers the influence of stop skipping action and bus capacity on the left-over passengers to determine the proper skipping stations for the bus fleet serving the entire route by minimizing both the waiting time and in-vehicle time of passengers and the running time of vehicles. A solution algorithm based on genetic algorithm is also presented to evaluate the degree of unbalanced passenger demand and optimize the limited-stop scheme. Then, the proper strategy is tested on a bus route in Changchun city of China. The threshold of degree assessment of unbalanced passenger demand can be calibrated and adapted to different passenger demands. PMID:29505585

Design of limited-stop service based on the degree of unbalance of passenger demand.

PubMed

Zhang, Hu; Zhao, Shuzhi; Liu, Huasheng; Liang, Shidong

2018-01-01

This paper presents a limited-stop service for a bus fleet to meet the unbalanced demand of passengers on a bus route and to improve the transit service of the bus route. This strategy includes two parts: a degree assessment of unbalanced passenger demand and an optimization of the limited-stop service. The degree assessment of unbalanced passenger demand, which is based on the different passenger demand between stations and the unbalance of passengers within the station, is used to judge whether implementing the limited-stop service is necessary for a bus route. The optimization of limited-stop service considers the influence of stop skipping action and bus capacity on the left-over passengers to determine the proper skipping stations for the bus fleet serving the entire route by minimizing both the waiting time and in-vehicle time of passengers and the running time of vehicles. A solution algorithm based on genetic algorithm is also presented to evaluate the degree of unbalanced passenger demand and optimize the limited-stop scheme. Then, the proper strategy is tested on a bus route in Changchun city of China. The threshold of degree assessment of unbalanced passenger demand can be calibrated and adapted to different passenger demands.

Secretion of the Intimin Passenger Domain Is Driven by Protein Folding*

PubMed Central

Leo, Jack C.; Oberhettinger, Philipp; Yoshimoto, Shogo; Udatha, D. B. R. K. Gupta; Morth, J. Preben; Schütz, Monika; Hori, Katsutoshi

2016-01-01

Intimin is an essential adhesin of attaching and effacing organisms such as entropathogenic Escherichia coli. It is also the prototype of type Ve secretion or inverse autotransport, where the extracellular C-terminal region or passenger is exported with the help of an N-terminal transmembrane β-barrel domain. We recently reported a stalled secretion intermediate of intimin, where the passenger is located in the periplasm but the β-barrel is already inserted into the membrane. Stalling of this mutant is due to the insertion of an epitope tag at the very N terminus of the passenger. Here, we examined how this insertion disrupts autotransport and found that it causes misfolding of the N-terminal immunoglobulin (Ig)-like domain D00. We could also stall the secretion by making an internal deletion in D00, and introducing the epitope tag into the second Ig-like domain, D0, also resulted in reduced passenger secretion. In contrast to many classical autotransporters, where a proximal folding core in the passenger is required for secretion, the D00 domain is dispensable, as the passenger of an intimin mutant lacking D00 entirely is efficiently exported. Furthermore, the D00 domain is slightly less stable than the D0 and D1 domains, unfolding at ∼200 piconewtons (pN) compared with ∼250 pN for D0 and D1 domains as measured by atomic force microscopy. Our results support a model where the secretion of the passenger is driven by sequential folding of the extracellular Ig-like domains, leading to vectorial transport of the passenger domain across the outer membrane in an N to C direction. PMID:27466361

Examining Passenger Flow Choke Points at Airports Using Discrete Event Simulation

NASA Technical Reports Server (NTRS)

Brown, Jeremy R.; Madhavan, Poomima

2011-01-01

The movement of passengers through an airport quickly, safely, and efficiently is the main function of the various checkpoints (check-in, security. etc) found in airports. Human error combined with other breakdowns in the complex system of the airport can disrupt passenger flow through the airport leading to lengthy waiting times, missing luggage and missed flights. In this paper we present a model of passenger flow through an airport using discrete event simulation that will provide a closer look into the possible reasons for breakdowns and their implications for passenger flow. The simulation is based on data collected at Norfolk International Airport (ORF). The primary goal of this simulation is to present ways to optimize the work force to keep passenger flow smooth even during peak travel times and for emergency preparedness at ORF in case of adverse events. In this simulation we ran three different scenarios: real world, increased check-in stations, and multiple waiting lines. Increased check-in stations increased waiting time and instantaneous utilization. while the multiple waiting lines decreased both the waiting time and instantaneous utilization. This simulation was able to show how different changes affected the passenger flow through the airport.

Ride quality criteria. [transportation system interior and passenger response to environment

NASA Technical Reports Server (NTRS)

Stephens, D. G.

1977-01-01

Ride quality refers to the interior or passenger environment of a transportation system as well as the passenger response to the environment. Ride quality factors are illustrated with the aid of a diagram presenting inputs to vehicle, the vehicle transfer function, the ride environment, the passenger response function, and the passenger ride response. The reported investigation considers the ride environment as measured on a variety of air and surface vehicles, the passenger response to the environment as determined from laboratory and field surveys, and criteria/standards for vibration, noise, and combined stimuli. Attention is given to the vertical vibration characteristics in cruise for aircraft and automobile, the aircraft vibration levels for various operating regimes, comparative noise levels during cruise, the discomfort level for a 9 Hz sinusoidal vibration, equal discomfort contours for vertical vibration, subjective response to noise in a speech situation, and noise and vibration levels for constant discomfort contours.

Intrastate passenger commuter ferry study : New Haven, Bridgeport, Norwalk, Stamford

DOT National Transportation Integrated Search

2001-03-01

This study analyzes the need and opportunity for establishing an Intrastate Passenger Commuter Ferry service along Long Island Sound, serving ports between Branford and Stamford, Connecticut. The feasibility of providing passenger ferry service focus...

Numerical analysis of air-flow and temperature field in a passenger car compartment

NASA Astrophysics Data System (ADS)

Kamar, Haslinda Mohamed; Kamsah, Nazri; Mohammad Nor, Ahmad Miski

2012-06-01

This paper presents a numerical study on the temperature field inside a passenger's compartment of a Proton Wira saloon car using computational fluid dynamics (CFD) method. The main goal is to investigate the effects of different glazing types applied onto the front and rear windscreens of the car on the distribution of air-temperature inside the passenger compartment in the steady-state conditions. The air-flow condition in the passenger's compartment is also investigated. Fluent CFD software was used to develop a three-dimensional symmetrical model of the passenger's compartment. Simplified representations of the driver and one rear passenger were incorporated into the CFD model of the passenger's compartment. Two types of glazing were considered namely clear insulated laminated tint (CIL) with a shading coefficient of 0.78 and green insulated laminate tint (GIL) with a shading coefficient of 0.5. Results of the CFD analysis were compared with those obtained when the windscreens are made up of clear glass having a shading coefficient of 0.86. Results of the CFD analysis show that for a given glazing material, the temperature of the air around the driver is slightly lower than the air around the rear passenger. Also, the use of GIL glazing material on both the front and rear windscreens significantly reduces the air temperature inside the passenger's compartment of the car. This contributes to a better thermal comfort condition to the occupants. Swirling air flow condition occurs in the passenger compartment. The air-flow intensity and velocity are higher along the side wall of the passenger's compartment compared to that along the middle section of the compartment. It was also found that the use of glazing materials on both the front and rear windscreen has no significant effects on the air-flow condition inside the passenger's compartment of the car.

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.

Research the simulation model of the passenger travel behavior in urban rail platform

NASA Astrophysics Data System (ADS)

Wang, Yujia; Yin, Xiangyong

2017-05-01

Based on the results of the research on the platform of the Beijing Chegongzhuang subway station in the line 2, the passenger travel behavior in urban rail platform is divided into 4 parts, which are the enter passenger walking, the passenger waiting distribution and queuing up before the door, passenger boarding and alighting and the alighting passengers walking, according to the social force model, simulation model was built based on Matlab software. Combined with the actual data of subway the Chegongzhuang subway station in the line 2, the simulation results show that the social force model is effective.

An investigation of passing stop signals at a passenger railroad

DOT National Transportation Integrated Search

2015-07-01

This report documents the findings and recommendations from an investigation into stop signal violations at an American passenger railroad. The investigation was requested by the Federal Railroad Administration (FRA) and the passenger railroad, and s...

Factors related to seatbelt-wearing among rear-seat passengers in Malaysia.

PubMed

Ng, Choy Peng; Law, Teik Hua; Wong, Shaw Voon; Kulanthayan, S

2013-01-01

The benefit of wearing a rear seatbelt in reducing the risk of motor vehicle crash-related fatalities and injuries has been well documented in previous studies. Wearing a seatbelt not only reduces the risk of injury to rear-seat passengers, but also reduces the risk of injury to front-seat occupant who could be crushed by unbelted rear-seat passengers in a motor vehicle crash. Despite the benefits of wearing a rear seatbelt, its rate of use in Malaysia is generally low. The objective of this study was to identify factors that are associated with the wearing of a seatbelt among rear-seat passengers in Malaysia. Multinomial logistic regression analysis of the results of a questionnaire survey of 1651 rear-seat passengers revealed that rear-seat passengers who were younger, male, single and less educated and who had a perception of a low level of legislation enforcement, a lower risk-aversion and less driving experience (only for passengers who are also drivers) were less likely to wear a rear seatbelt. There was also a significant positive correlation between driver seatbelt and rear seatbelt-wearing behaviour. This implies that, in regards to seatbelt-wearing behaviour, drivers are more likely to adopt the same seatbelt-wearing behaviour when travelling as rear-seat passengers as they do when driving. These findings are crucial to the development of new interventions to increase the compliance rate of wearing a rear seatbelt. Copyright © 2012 Elsevier Ltd. All rights reserved.

The effect of passengers on teen driver behavior : traffic tech.

DOT National Transportation Integrated Search

2012-04-01

A number of studies have shown that passengers substantially : increase the risk of crashes for young, novice drivers. : This increased risk may result from distractions that young : passengers create for drivers. Alternatively, the presence : of pas...

Intercity passenger rail : issues associated with a possible Amtrak liquidation

DOT National Transportation Integrated Search

1998-03-01

The National Railroad Passenger Corporation (Amtrak) carries over 20 million : passengers a year through the trains that it operates. It also provides services, : such as dispatching, to state and local commuter rail operators. Since 1971, : Amtrak h...

49 CFR 39.53 - What information must PVOs provide to passengers with a disability?

Code of Federal Regulations, 2010 CFR

2010-10-01

... 49 Transportation 1 2010-10-01 2010-10-01 false What information must PVOs provide to passengers with a disability? 39.53 Section 39.53 Transportation Office of the Secretary of Transportation TRANSPORTATION FOR INDIVIDUALS WITH DISABILITIES: PASSENGER VESSELS Information for Passengers § 39.53 What information must PVOs provide to passengers...

49 CFR 39.53 - What information must PVOs provide to passengers with a disability?

Code of Federal Regulations, 2011 CFR

2011-10-01

... 49 Transportation 1 2011-10-01 2011-10-01 false What information must PVOs provide to passengers with a disability? 39.53 Section 39.53 Transportation Office of the Secretary of Transportation TRANSPORTATION FOR INDIVIDUALS WITH DISABILITIES: PASSENGER VESSELS Information for Passengers § 39.53 What information must PVOs provide to passengers...

49 CFR 39.53 - What information must PVOs provide to passengers with a disability?

Code of Federal Regulations, 2014 CFR

2014-10-01

... 49 Transportation 1 2014-10-01 2014-10-01 false What information must PVOs provide to passengers with a disability? 39.53 Section 39.53 Transportation Office of the Secretary of Transportation TRANSPORTATION FOR INDIVIDUALS WITH DISABILITIES: PASSENGER VESSELS Information for Passengers § 39.53 What information must PVOs provide to passengers...

49 CFR 39.53 - What information must PVOs provide to passengers with a disability?

Code of Federal Regulations, 2012 CFR

2012-10-01

... 49 Transportation 1 2012-10-01 2012-10-01 false What information must PVOs provide to passengers with a disability? 39.53 Section 39.53 Transportation Office of the Secretary of Transportation TRANSPORTATION FOR INDIVIDUALS WITH DISABILITIES: PASSENGER VESSELS Information for Passengers § 39.53 What information must PVOs provide to passengers...

Aircraft passenger comfort experience: underlying factors and differentiation from discomfort.

PubMed

Ahmadpour, Naseem; Robert, Jean-Marc; Lindgaard, Gitte

2016-01-01

Previous studies defined passengers' comfort based on their concerns during the flight and a set of eight experiential factors such as 'peace of mind', 'physical wellbeing', 'pleasure', etc. One Objective of this paper was to determine whether the factors underlying the passengers' experience of comfort differ from those of discomfort. Another objective was to cross-validate those factors. In the first study, respondents provided written reports of flight comfort and discomfort experiences separately and gave ratings on the impact of the eight factors on each experience. Follow up interviews were also conducted. Significant difference was found between comfort and discomfort ratings for two factors of 'pleasure', denoted by one's concern for stimulation, ambience and exceeded expectations, and 'physical wellbeing' characterized in terms of bodily support and energy. However, there were no significant differences between the comfort and discomfort ratings on the other six factors. The evidence does not support the proposition that passenger comfort and discomfort are underline by different sets of factors. It is therefore suggested that the evaluation of overall passenger comfort experience, as a whole, employ one spectrum ranging from extreme comfort to discomfort. In study two, a pool of comfort descriptors was collected. Those that were less relevant to passenger comfort were eliminated in a number of steps. Factor analysis was used to classify the remaining descriptors, using respondents' ratings on their potential impact on passenger comfort. Seven factors corresponded to the pre-determined passenger comfort factors from previous research, validating those with an exception of 'proxemics' (concerning one's privacy and control over their situation) but it was argued that this is due to the nature of the factor itself, which is context dependent and generally perceived unconsciously. Copyright © 2015 Elsevier Ltd and The Ergonomics Society. All rights reserved.

Effects of cold exposure on wet aircraft passengers : a review.

DOT National Transportation Integrated Search

1994-05-01

The incorporation of a cabin water spray system (CWSS) aboard commercial passenger aircraft has been suggested as a mechanism of reducing passenger death and injury from the fire and smoke commonly associated with aircraft accidents. A potential heal...

Forecasting the Short-Term Passenger Flow on High-Speed Railway with Neural Networks

PubMed Central

Xie, Mei-Quan; Li, Xia-Miao; Zhou, Wen-Liang; Fu, Yan-Bing

2014-01-01

Short-term passenger flow forecasting is an important component of transportation systems. The forecasting result can be applied to support transportation system operation and management such as operation planning and revenue management. In this paper, a divide-and-conquer method based on neural network and origin-destination (OD) matrix estimation is developed to forecast the short-term passenger flow in high-speed railway system. There are three steps in the forecasting method. Firstly, the numbers of passengers who arrive at each station or depart from each station are obtained from historical passenger flow data, which are OD matrices in this paper. Secondly, short-term passenger flow forecasting of the numbers of passengers who arrive at each station or depart from each station based on neural network is realized. At last, the OD matrices in short-term time are obtained with an OD matrix estimation method. The experimental results indicate that the proposed divide-and-conquer method performs well in forecasting the short-term passenger flow on high-speed railway. PMID:25544838

Forecasting the short-term passenger flow on high-speed railway with neural networks.

PubMed

Xie, Mei-Quan; Li, Xia-Miao; Zhou, Wen-Liang; Fu, Yan-Bing

2014-01-01

Short-term passenger flow forecasting is an important component of transportation systems. The forecasting result can be applied to support transportation system operation and management such as operation planning and revenue management. In this paper, a divide-and-conquer method based on neural network and origin-destination (OD) matrix estimation is developed to forecast the short-term passenger flow in high-speed railway system. There are three steps in the forecasting method. Firstly, the numbers of passengers who arrive at each station or depart from each station are obtained from historical passenger flow data, which are OD matrices in this paper. Secondly, short-term passenger flow forecasting of the numbers of passengers who arrive at each station or depart from each station based on neural network is realized. At last, the OD matrices in short-term time are obtained with an OD matrix estimation method. The experimental results indicate that the proposed divide-and-conquer method performs well in forecasting the short-term passenger flow on high-speed railway.

Passengers' perception of the safety demonstration on board an aircraft

NASA Astrophysics Data System (ADS)

Ruenruoy, Ratchada

The cabin safety demonstration on board an aircraft is one of the methods to provide safety information for passengers before aircraft takeoff. However, passengers' enthusiasm toward safety demonstrations is normally low. Therefore, the study of passengers' perception toward safety briefings on board an aircraft is important in increasing the safety awareness for the travelling public on commercial aircraft. A survey was distributed to measure the perceptions of Middle Tennessee State University (MTSU) faculty and staff, Aerospace students, and international students who have traveled in the last year. It was generally found that watching the cabin safety demonstration before aircraft takeoff was believed to be important for passengers. However, the attention to the safety demonstration remained low because the safety briefings were not good enough in terms of clear communication, particularly in the recorded audio demonstration and the live safety demonstration methods of briefing.

14 CFR 23.853 - Passenger and crew compartment interiors.

Code of Federal Regulations, 2010 CFR

2010-01-01

... constructed of at least fire resistant materials and must contain fires likely to occur in it under normal use... Design and Construction Fire Protection § 23.853 Passenger and crew compartment interiors. For each compartment to be used by the crew or passengers: (a) The materials must be at least flame-resistant; (b...

46 CFR 71.75-5 - Passenger Ship Safety Certificate.

Code of Federal Regulations, 2010 CFR