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Sample records for junonia coenia densovirus

  1. Improved injection needles facilitate germline transformation of the buckeye butterfly Junonia coenia.

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    Beaudette, Kahlia; Hughes, Tia M; Marcus, Jeffrey M

    2014-01-01

    Germline transformation with transposon vectors is an important tool for insect genetics, but progress in developing transformation protocols for butterflies has been limited by high post-injection ova mortality. Here we present an improved glass injection needle design for injecting butterfly ova that increases survival in three Nymphalid butterfly species. Using the needles to genetically transform the common buckeye butterfly Junonia coenia, the hatch rate for injected Junonia ova was 21.7%, the transformation rate was 3%, and the overall experimental efficiency was 0.327%, a substantial improvement over previous results in other butterfly species. Improved needle design and a higher efficiency of transformation should permit the deployment of transposon-based genetic tools in a broad range of less fecund lepidopteran species.

  2. Pathogenicity of diatraea saccharalis Densovirus to Host Insets and Characterization of its Viral Genome

    Institute of Scientific and Technical Information of China (English)

    Nazaire Kouassi; Jian-xin PENG; Yi LI; Cristina Cavallaro; Jean-Claude Veyrunes; Max Bergoin

    2007-01-01

    Pathogenicity of the Diatraea saccharalis densovirus (DsDNV) was tested on its host larvae.The results showed that up to 4 days after inoculation,no larvae mortality was observed and the infected larvae started to exhibit the infection symptoms from the fourth day.After 5 days of infection,the cumulative mortality of infected larvae increased significantly and reached 60% after 12 days and 100% after 21 days of infection,whereas that of the control group was only 10% and 20%,respectively,after same periods of infection,suggesting that the high mortality of infected larvae groups was due to the high pathogenicity of DsDNV.The size of the DsDNA was determined by Electron microscopy visualization of viral DNA molecules and gel electrophoresis of both native and endonuclease digested DNA fragments.The total length of the native DsDNA was about 5.95 kb.The DsDNV DNA was digested with 16 restriction enzymes and a restriction map of those enzymes was constructed with 41 restriction sites.Comparison of the restriction map of the DsDNV genome with those of the genomes ofJunonia coenia densovirus (JcDNV) and Galleria mellonella densovirus (GmDNV) indicated that the three densovirus genomes were found to share many identical restriction sites.Thus,most of the restriction sites of the following endonucleases Bam H Ⅰ,Hha Ⅰ,Xba Ⅰ,Cla Ⅰ,Asp 700,Spe Ⅰ,Nco Ⅰ and Bcl Ⅰ,were found to be conserved among the three densovirus genomes.Symmetrical cleavage sites mapped at the both ends of the genome suggested the presence of inverted terminal repeats (ITRs) whose size was estimated to be about 500 bp.The similar genome size,almost identical restriction sites and presence of an ITR of about 500 bp for these three densoviruses suggested that they belong to the same group of ambisense densoviruses.

  3. Insect cell transformation vectors that support high level expression and promoter assessment in insect cell culture

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    A somatic transformation vector, pDP9, was constructed that provides a simplified means of producing permanently transformed cultured insect cells that support high levels of protein expression of foreign genes. The pDP9 plasmid vector incorporates DNA sequences from the Junonia coenia densovirus th...

  4. Molecular Population Structure of Junonia Butterflies from French Guiana, Guadeloupe, and Martinique

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    Amber P. Gemmell

    2014-01-01

    Full Text Available Up to 9 described species of Junonia butterflies occur in the Americas, but authorities disagree due to species similarities, geographical and seasonal variability, and possible hybridization. In dispute is whether Caribbean Junonia are conspecific with South American species. Cytochrome oxidase I (COI barcodes, wingless (wg sequences, and Randomly Amplified Fingerprints (RAF were studied to reveal Junonia population structure in French Guiana, Guadeloupe, Martinique, and Argentina. Phylogenetic analysis of COI recovered 2 haplotype groups, but most Junonia species can have either haplotype, so COI barcodes are ambiguous. Analysis of nuclear wingless alleles revealed geographic patterns but did not identify Junonia species. Nuclear RAF genotyping distinguished 11 populations of Junonia arranged into 3 clusters. Gene flow occurs within clusters but is limited between clusters. One cluster included all Argentinian samples. Two clusters included samples from French Guiana, Martinique, and Guadeloupe and appear to be divided by larval host plant use (Lamiales versus Scrophulariales. Many Junonia taxa were distributed across populations, possibly reflecting patterns of genetic exchange. We had difficulty distinguishing between the Caribbean forms J. zonalis and J. neildi, but we demonstrate that Caribbean Junonia are genetically distinct from South American J. evarete and J. genoveva, supporting the taxonomic hypothesis that they are heterospecific.

  5. Comparison of mosquito densoviruses: two clades of viruses isolated from indigenous mosquitoes.

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    Sangdee, Kusavadee; Pattanakitsakul, Sa-Nga

    2013-07-04

    We analyzed the phylogenetic tree of densoviruses isolated from indigenous mosquitoes and mosquito cell lines. Our findings suggest two distinct clades of densovirus. The viruses in the first clade were isolated from an indigenous mosquito which had the Aedes aegypti densovirus (AaeDNV) as a representative virus. The other clade of viruses was isolated from mosquito indigenous cell line which had the Aedes albopictus densovirus (AalDNV) as the representative virus. The origin of the two clades of DNVs is unclear but the phylogenetic trees were significantly different from each other. The two major densoviruses, AaeDNV and AalDNV, that infect mosquitoes that are known to carry viruses responsible for dengue hemorrhagic fever and yellow fever. Understanding the evolution of these two clades of densoviruses is important for studying the distribution of these viruses in mosquito cell lines and the information gained may be applied to understanding other viruses in various mosquito cell lines.

  6. Complete Genome Sequence of a Putative Densovirus of the Asian Citrus Psyllid, Diaphorina citri.

    Science.gov (United States)

    Nigg, Jared C; Nouri, Shahideh; Falk, Bryce W

    2016-07-28

    Here, we report the complete genome sequence of a putative densovirus of the Asian citrus psyllid, Diaphorina citri Diaphorina citri densovirus (DcDNV) was originally identified through metagenomics, and here, we obtained the complete nucleotide sequence using PCR-based approaches. Phylogenetic analysis places DcDNV between viruses of the Ambidensovirus and Iteradensovirus genera.

  7. Structure of Penaeus stylirostris Densovirus, a Shrimp Pathogen

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    Kaufmann, Bärbel; Bowman, Valorie D.; Li, Yi; Szelei, Jozsef; Waddell, Peter J.; Tijssen, Peter; Rossmann, Michael G. (INRS); (Purdue)

    2010-11-16

    Penaeus stylirostris densovirus (PstDNV), a pathogen of penaeid shrimp, causes significant damage to farmed and wild shrimp populations. In contrast to other parvoviruses, PstDNV probably has only one type of capsid protein that lacks the phospholipase A2 activity that has been implicated as a requirement during parvoviral host cell infection. The structure of recombinant virus-like particles, composed of 60 copies of the 37.5-kDa coat protein, the smallest parvoviral capsid protein reported thus far, was determined to 2.5-{angstrom} resolution by X-ray crystallography. The structure represents the first near-atomic resolution structure within the genus Brevidensovirus. The capsid protein has a {beta}-barrel 'jelly roll' motif similar to that found in many icosahedral viruses, including other parvoviruses. The N-terminal portion of the PstDNV coat protein adopts a 'domain-swapped' conformation relative to its twofold-related neighbor similar to the insect parvovirus Galleria mellonella densovirus (GmDNV) but in stark contrast to vertebrate parvoviruses. However, most of the surface loops have little structural resemblance to any of the known parvoviral capsid proteins.

  8. Illumina based whole mitochondrial genome of Junonia iphita reveals minor intraspecific variation

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    Catherine Vanlalruati

    2015-12-01

    Full Text Available In the present study, the near complete mitochondrial genome (mitogenome of Junonia iphita (Lepidoptera: Nymphalidae: Nymphalinae was determined to be 14,892 bp. The gene order and orientation are identical to those in other butterfly species. The phylogenetic tree constructed from the whole mitogenomes using the 13 protein coding genes (PCGs defines the genetic relatedness of the two J. iphita species collected from two different regions. All the Junonia species clustered together, and were further subdivided into clade one consisting of J. almana and J. orithya and clade two comprising of the two J. iphita which were collected from Indo and Indochinese subregions separated by river barrier. Comparison between the two J. iphita sequences revealed minor variations and Single Nucleotide Polymorphisms were identified at 51 sites amounting to 0.4% of the entire mitochondrial genome.

  9. Genetic and biological characterization of a densovirus isolate that affects dengue virus infection

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    Ana Luiza Pamplona Mosimann

    2011-05-01

    Full Text Available Brevidensoviruses have an encapsidated, single-stranded DNA genome that predominantly has a negative polarity. In recent years, they have received particular attention due to their potential role in the biological control of pathogenic arboviruses and to their unnoticed presence in cell cultures as contaminants. In addition, brevidensoviruses may also be useful as viral vectors. This study describes the first genetic and biological characterization of a mosquito densovirus that was isolated in Brazil; moreover, we examined the phylogenetic relationship between this isolate and the other brevidensoviruses. We further demonstrate that this densovirus has the potential to be used to biologically control dengue virus (DENV infection with in vitro co-infection experiments. The present study provides evidence that this densovirus isolate is a fast-spreading virus that affects cell growth and DENV infection.

  10. Discovery of urchin-associated densoviruses (family Parvoviridae) in coastal waters of the Big Island, Hawaii.

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    Gudenkauf, Brent M; Eaglesham, James B; Aragundi, William M; Hewson, Ian

    2014-03-01

    Echinoderms are important constituents of marine ecosystems, where they may influence the recruitment success of benthic flora and fauna, and are important consumers of detritus and plant materials. There are currently no described viruses of echinoderms. We used a viral metagenomic approach to examine viral consortia within three urchins - Colobocentrotus atratus, Tripneustes gratilla and Echinometra mathaei - which are common constituents of reef communities in the Hawaiian archipelago. Metagenomic libraries revealed the presence of bacteriophages and densoviruses (family Parvoviridae) in tissues of all three urchins. Densoviruses are known typically to infect terrestrial and aquatic arthropods. Urchin-associated densoviruses were detected by quantitative PCR in all tissues tested, and were also detected in filtered suspended matter (>0.2 µm) from plankton and in sediments at several locations near to where the urchins were collected for metagenomic analysis. To the best of our knowledge, this is the first report of echinoderm-associated viruses, which extends the known host range of parvoviruses.

  11. Structure comparisons of Aedes albopictus densovirus with other parvoviruses

    Institute of Scientific and Technical Information of China (English)

    2007-01-01

    Parvoviridae is a family of the smallest viruses known with a wide variety of hosts. The capsid structure of the Aedes albopictus C6/36 cell densovirus (C6/36 DNV) at 1.2-nm resolution was obtained by elec-tron cryomicroscopy (cryoEM) and three-dimensional (3D) image reconstruction. Structure compari-sons between the C6/36 DNV and other parvoviruses reveal that the degree of structural similarity be-tween C6/36 DNV and the human parvovirus B19 is higher than that between C6/36 DNV and other in-sect parvoviruses. The amino acid sequence comparisons of structural and non-structural proteins also reveal higher levels of similarity between C6/36 DNV and parvovirus B19 than those between C6/36 DNV and other parvoviruses. These findings indicate that C6/36 DNV is closely related to the human virus B19, and the former might evolve from the human species other than from other insect viruses.

  12. Structure comparisons of Aedes albopictus densovirus with other parvoviruses

    Institute of Scientific and Technical Information of China (English)

    CHENG LingPeng; CHEN SenXiong; Z. H. ZHOU; ZHANG JingQiang

    2007-01-01

    Parvoviridae is a family of the smallest viruses known with a wide variety of hosts. The capsid structure of the Aedes albopictus C6/36 cell densovirus (C6/36 DNV) at 1.2-nm resolution was obtained by electron cryomicroscopy (cryoEM) and three-dimensional (3D) image reconstruction. Structure comparisons between the C6/36 DNV and other parvoviruses reveal that the degree of structural similarity between C6/36 DNV and the human parvovirus B19 is higher than that between C6/36 DNV and other insect parvoviruses. The amino acid sequence comparisons of structural and non-structural proteins also reveal higher levels of similarity between C6/36 DNV and parvovirus B19 than those between C6/36 DNV and other parvoviruses. These findings indicate that C6/36 DNV is closely related to the human virus B19, and the former might evolve from the human species other than from other insect viruses.

  13. Complete mitochondrial genome of Chocolate Pansy, Junonia iphita (Lepidoptera: Nymphalidae: Nymphalinae).

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    Vanlalruati, Catherine; Mandal, Surajit De; Gurusubramanian, Guruswami; Senthil Kumar, Nachimuthu

    2016-07-01

    The complete mitochondrial genome of Junonia iphita was determined to be 15,433 bp in length, including 37 typical mitochondrial genes and an AT-rich region. All the protein coding genes (PCGs) are initiated by typical ATN codons, except cox1 gene that is by CGA codon. Eight genes use complete termination codon (TAA), whereas the cox1, cox2 and nad5 genes end with single T; nad4 and nad1 ends with stop codon TA. All the tRNA show secondary cloverleaf structures except trnS1 (AGN). The A + T rich region is 546 bp in length containing ATAGA motif followed by a 18 bp poly-T stretch, two microsatellite-like (TA)9 elements and 8 bp poly-A stretch immediately upstream of trnM gene.

  14. Three-dimensional structure determination of capsid of Aedes albopicus C6/36 cell densovirus

    Institute of Scientific and Technical Information of China (English)

    CHENG Lingpeng; CHEN Senxiong; Jenifer M.Brannan; Joanita Jakana; ZHANG Qinfen; Z.H.Zhou; ZHANG Jingqiang

    2004-01-01

    The three-dimensional structure of capsid of Aedes albopictus C6/36 densovirus was determined to 14-(A) resolution by electron cryomicroscopy and computer reconstruction. The triangulation number of the capsid is 1. There are 12 holes in each triangular face and a spike on each 5-fold vertex. The validity of the capsid and nucleic acid densities in the reconstructions was discussed.

  15. Insights into Population Origins of Neotropical Junonia (Lepidoptera: Nymphalidae: Nymphalinae Based on Mitochondrial DNA

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    Edward Pfeiler

    2012-01-01

    Full Text Available Cytochrome c oxidase subunit I (COI sequences were used to estimate demographic histories of populations of the buckeye butterfly Junonia genoveva (Cramer from Costa Rica and Mexico. Previous studies have revealed significant structure between populations of J. genoveva from coastal regions of northwestern Mexico, which utilize black mangrove Avicennia germinans (Acanthaceae as a larval host plant, and inland populations from Costa Rica that feed on different hosts in the families Acanthaceae and Verbenaceae. The Mexico population of J. genoveva reported on here is located near the Northern limit of black mangrove habitat on the Pacific coast of North America and is hypothesized to have been established by northward migrations and colonization from southern source populations. The mismatch distribution, Bayesian skyline analyses, and maximum likelihood analyses carried out in FLUCTUATE were used to estimate changes in female effective population size (Nef over time in the two populations. Differences found in COI haplotype diversity, present-day Nef, and the timing of population expansions are consistent with the hypothesis that the Mexico population of J. genoveva is the more recently evolved.

  16. Evidence for the Deflective Function of Eyespots in Wild Junonia evarete Cramer (Lepidoptera, Nymphalidae).

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    Pinheiro, C E G; Antezana, M A; Machado, L P

    2014-02-01

    Junonia evarete Cramer is a fast-flying butterfly that perches on the ground with wings opened exhibiting four eyespots close to wing borders. These eyespots presumably function either to intimidate predators, like insectivorous birds, or to deflect bird attacks to less vital parts of the body. We assessed the form, frequency, and location of beak marks on the wings of wild butterflies in central Brazil during two not consecutive years. We found that almost 50% of males and 80% of females bore signals of predator attacks (wing tears), most of them consisting of partially or totally V-shaped forms apparently produced by birds. Males were significantly less attacked and showed a lower proportion of attacks on eyespots than females, suggesting they are better to escape bird attacks. In contrast, females were heavily attacked on eyespots. Eyespot tears in females were higher (and significant different) than expected by chance, indicating that birds do attempt to reach the eyespots when striking on these butterflies. Other comparisons involving the proportion of tears directed or not directed to eyespots in males and females are presented and discussed.

  17. Densovirus is a mutualistic symbiont of a global crop pest (Helicoverpa armigera and protects against a baculovirus and Bt biopesticide.

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    Pengjun Xu

    2014-10-01

    Full Text Available Mutualistic associations between symbiotic bacteria and their hosts are common within insect systems. However, viruses are often considered as pathogens even though some have been reported to be beneficial to their hosts. Herein, we report a novel densovirus, Helicoverpa armigera densovirus-1 (HaDNV-1 that appears to be beneficial to its host. HaDNV-1 was found to be widespread in wild populations of H. armigera adults (>67% prevalence between 2008 and 2012. In wild larval populations, there was a clear negative interaction between HaDNV-1 and H. armigera nucleopolyhedrovirus (HaNPV, a baculovirus that is widely used as a biopesticide. Laboratory bioassays revealed that larvae hosting HaDNV-1 had significantly enhanced resistance to HaNPV (and lower viral loads, and that resistance to Bacillus thuringiensis (Bt toxin was also higher at low doses. Laboratory assays indicated that the virus was mainly distributed in the fat body, and could be both horizontally- and vertically-transmitted, though the former occurred only at large challenge doses. Densovirus-positive individuals developed more quickly and had higher fecundity than uninfected insects. We found no evidence for a negative effect of HaDNV-1 infection on H. armigera fitness-related traits, strongly suggesting a mutualistic interaction between the cotton bollworm and its densovirus.

  18. Densovirus associated with sea-star wasting disease and mass mortality.

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    Hewson, Ian; Button, Jason B; Gudenkauf, Brent M; Miner, Benjamin; Newton, Alisa L; Gaydos, Joseph K; Wynne, Janna; Groves, Cathy L; Hendler, Gordon; Murray, Michael; Fradkin, Steven; Breitbart, Mya; Fahsbender, Elizabeth; Lafferty, Kevin D; Kilpatrick, A Marm; Miner, C Melissa; Raimondi, Peter; Lahner, Lesanna; Friedman, Carolyn S; Daniels, Stephen; Haulena, Martin; Marliave, Jeffrey; Burge, Colleen A; Eisenlord, Morgan E; Harvell, C Drew

    2014-12-01

    Populations of at least 20 asteroid species on the Northeast Pacific Coast have recently experienced an extensive outbreak of sea-star (asteroid) wasting disease (SSWD). The disease leads to behavioral changes, lesions, loss of turgor, limb autotomy, and death characterized by rapid degradation ("melting"). Here, we present evidence from experimental challenge studies and field observations that link the mass mortalities to a densovirus (Parvoviridae). Virus-sized material (i.e., <0.2 μm) from symptomatic tissues that was inoculated into asymptomatic asteroids consistently resulted in SSWD signs whereas animals receiving heat-killed (i.e., control) virus-sized inoculum remained asymptomatic. Viral metagenomic investigations revealed the sea star-associated densovirus (SSaDV) as the most likely candidate virus associated with tissues from symptomatic asteroids. Quantification of SSaDV during transmission trials indicated that progression of SSWD paralleled increased SSaDV load. In field surveys, SSaDV loads were more abundant in symptomatic than in asymptomatic asteroids. SSaDV could be detected in plankton, sediments and in nonasteroid echinoderms, providing a possible mechanism for viral spread. SSaDV was detected in museum specimens of asteroids from 1942, suggesting that it has been present on the North American Pacific Coast for at least 72 y. SSaDV is therefore the most promising candidate disease agent responsible for asteroid mass mortality.

  19. Factors influencing infection and transmission of Anopheles gambiae densovirus (AgDNV) in mosquitoes.

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    Barik, Tapan K; Suzuki, Yasutsugu; Rasgon, Jason L

    2016-01-01

    Anopheles gambiae densovirus (AgDNV) is a potential microbial agent for paratransgenesis and gene transduction in An. gambiae, the major vector of human malaria in sub-Saharan Africa. Understanding the interaction between AgDNV and An. gambiae is critical for using AgDNV in a basic and applied manner for Anopheles gene manipulation. Here, we tested the effects of mosquito age, sex, blood feeding status, and potential for horizontal transmission using an enhanced green fluorescent protein (EGFP) reporter AgDNV system. Neither mosquito age at infection nor feeding regime affected viral titers. Female mosquitoes were more permissive to viral infection than males. Despite low viral titers, infected males were able to venereally transmit virus to females during mating, where the virus was localized with the transferred sperm in the spermathecae. These findings will be useful for designing AgDNV-based strategies to manipulate Anopheles gambiae.

  20. Factors influencing infection and transmission of Anopheles gambiae densovirus (AgDNV in mosquitoes

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    Tapan K. Barik

    2016-11-01

    Full Text Available Anopheles gambiae densovirus (AgDNV is a potential microbial agent for paratransgenesis and gene transduction in An. gambiae, the major vector of human malaria in sub-Saharan Africa. Understanding the interaction between AgDNV and An. gambiae is critical for using AgDNV in a basic and applied manner for Anopheles gene manipulation. Here, we tested the effects of mosquito age, sex, blood feeding status, and potential for horizontal transmission using an enhanced green fluorescent protein (EGFP reporter AgDNV system. Neither mosquito age at infection nor feeding regime affected viral titers. Female mosquitoes were more permissive to viral infection than males. Despite low viral titers, infected males were able to venereally transmit virus to females during mating, where the virus was localized with the transferred sperm in the spermathecae. These findings will be useful for designing AgDNV-based strategies to manipulate Anopheles gambiae.

  1. Densovirus associated with sea-star wasting disease and mass mortality

    Science.gov (United States)

    Hewson, Ian; Button, Jason B.; Gudenkauf, Brent M.; Miner, Benjamin; Newton, Alisa L.; Gaydos, Joseph K.; Wynne, Janna; Groves, Cathy L.; Hendler, Gordon; Murray, Michael; Fradkin, Steven; Breitbart, Mya; Fahsbender, Elizabeth; Lafferty, Kevin D.; Kilpatrick, A. Marm; Miner, C. Melissa; Raimondi, Peter T.; Lahner, Lesanna L.; Friedman, Carolyn S.; Danielson, Stephen D.; Haulena, Martin; Marliave, Jeffrey; Burge, Colleen A.; Eisenlord, Morgan E.; Harvell, C. Drew

    2015-01-01

    Populations of at least 20 asteroid species on the Northeast Pacific Coast have recently experienced an extensive outbreak of sea-star (asteroid) wasting disease (SSWD). The disease leads to behavioral changes, lesions, loss of turgor, limb autotomy, and death characterized by rapid degradation (“melting”). Here, we present evidence from experimental challenge studies and field observations that link the mass mortalities to a densovirus (Parvoviridae). Virus-sized material (i.e., echinoderms, providing a possible mechanism for viral spread. SSaDV was detected in museum specimens of asteroids from 1942, suggesting that it has been present on the North American Pacific Coast for at least 72 y. SSaDV is therefore the most promising candidate disease agent responsible for asteroid mass mortality.

  2. Out-of-Africa origin and dispersal-mediated diversification of the butterfly genus Junonia (Nymphalidae: Nymphalinae).

    Science.gov (United States)

    Kodandaramaiah, U; Wahlberg, N

    2007-11-01

    The relative importance of dispersal and vicariance in the diversification of taxa has been much debated. Within butterflies, a few studies published so far have demonstrated vicariant patterns at the global level. We studied the historical biogeography of the genus Junonia (Nymphalidae: Nymphalinae) at the intercontinental level based on a molecular phylogeny. The genus is distributed over all major biogeographical regions of the world except the Palaearctic. We found dispersal to be the dominant process in the diversification of the genus. The genus originated and started diversifying in Africa about 20 Ma and soon after dispersed into Asia possibly through the Arabian Peninsula. From Asia, there were dispersals into Africa and Australasia, all around 5 Ma. The origin of the New World species is ambiguous; the ancestral may have dispersed from Asia via the Beringian Strait or from Africa over the Atlantic, about 3 Ma. We found no evidence for vicariance at the intercontinental scale. We argue that dispersal is as important as vicariance, if not more, in the global diversification of butterflies.

  3. Expression of non-structural protein NS3 gene of Bombyx mori densovirus (China isolate)

    Institute of Scientific and Technical Information of China (English)

    Huijuan Yin; Qin Yao; Zhongjian Guo; Fang Bao; Wei Yu; Jun Li; Keping Chen

    2008-01-01

    The invertebrate parvovirus Bombyx mori Densonucleosis Virus type 3 (China isolate),named BmDNV-3,is a kind of bidensovirus.It is a new type of virus with unique replication mechanisms.To investigate the effects of the NS3 gene during viral DNA replication,a pair of primers was designed for amplifying NS3 gene of Bombyx mori densovirus (China isolate).Gene NS3 amplified was cloned into a prokaryotic expression vector pET-30a and the donor plasmid pFastBacHTe,respectively.The NS3 protein was expressed in Escherichia coli BL21.The pFastBacHTe-NS3 was transformed to E.coli DH10Bac.The recombinant bacmid baculoviruses (rBacmid-EGFP-NS3)isolated from the white colonies were transfected into BraN-4 cells using a transfection reagent.BmN-4 cells were infected with recom-binant virus to express fusion proteins.The expression of fusion protein around 30 kDa in E.coli BL21 was identified by SDS-PAGE,Western blotting,and mass spectrometry.The expressed NS3 protein by B.mor/nucleopolyhedrovirus bacmid system was confirmed byWestern blotting using an anti-NS3 polyclonal antibody.And about 45 kDa protein was found.The expressed fusion protein was smalleithan the expected size of EGFP-NS3,55 kDa.Western blotting analysis indicated that EGFP-NS3 protein was expressed in infected lar-vae with smaller molecular size.

  4. Development of non-defective recombinant densovirus vectors for microRNA delivery in the invasive vector mosquito, Aedes albopictus

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    Liu, Peiwen; Li, Xiaocong; Gu, Jinbao; Dong, Yunqiao; Liu, Yan; Santhosh, Puthiyakunnon; Chen, Xiaoguang

    2016-01-01

    We previously reported that mosquito densoviruses (MDVs) are potential vectors for delivering foreign nucleic acids into mosquito cells. However, considering existing expression strategies, recombinant viruses would inevitably become replication-defective viruses and lose their ability for secondary transmission. The packaging limitations of the virion represent a barrier for the development of MDVs for viral paratransgenesis or as high-efficiency bioinsecticides. Herein, we report the development of a non-defective recombinant Aedes aegypti densovirus (AaeDV) miRNA expression system, mediated by an artificial intron, using an intronic miRNA expression strategy. We demonstrated that this recombinant vector could be used to overexpress endogenous miRNAs or to decrease endogenous miRNAs by generating antisense sponges to explore the biological functions of miRNAs. In addition, the vector could express antisense-miRNAs to induce efficient gene silencing in vivo and in vitro. The recombinant virus effectively self-replicated and retained its secondary transmission ability, similar to the wild-type virus. The recombinant virus was also genetically stable. This study demonstrated the first construction of a non-defective recombinant MDV miRNA expression system, which represents a tool for the functional analysis of mosquito genes and lays the foundation for the application of viral paratransgenesis for dengue virus control. PMID:26879823

  5. Development of non-defective recombinant densovirus vectors for microRNA delivery in the invasive vector mosquito, Aedes albopictus.

    Science.gov (United States)

    Liu, Peiwen; Li, Xiaocong; Gu, Jinbao; Dong, Yunqiao; Liu, Yan; Santhosh, Puthiyakunnon; Chen, Xiaoguang

    2016-02-16

    We previously reported that mosquito densoviruses (MDVs) are potential vectors for delivering foreign nucleic acids into mosquito cells. However, considering existing expression strategies, recombinant viruses would inevitably become replication-defective viruses and lose their ability for secondary transmission. The packaging limitations of the virion represent a barrier for the development of MDVs for viral paratransgenesis or as high-efficiency bioinsecticides. Herein, we report the development of a non-defective recombinant Aedes aegypti densovirus (AaeDV) miRNA expression system, mediated by an artificial intron, using an intronic miRNA expression strategy. We demonstrated that this recombinant vector could be used to overexpress endogenous miRNAs or to decrease endogenous miRNAs by generating antisense sponges to explore the biological functions of miRNAs. In addition, the vector could express antisense-miRNAs to induce efficient gene silencing in vivo and in vitro. The recombinant virus effectively self-replicated and retained its secondary transmission ability, similar to the wild-type virus. The recombinant virus was also genetically stable. This study demonstrated the first construction of a non-defective recombinant MDV miRNA expression system, which represents a tool for the functional analysis of mosquito genes and lays the foundation for the application of viral paratransgenesis for dengue virus control.

  6. Restoration of an endangered plant, Hygrophila pogonocalyx, leads to an adaptive host shift of the chocolate pansy (Junonia iphita iphita).

    Science.gov (United States)

    Tan, Wen-Hao; Liu, Tsui-Hua; Lin, Y Kirk; Hsu, Yu-Feng

    2014-08-01

    Anthropogenic introduction of a plant species may cause novel encounters between the plant and local herbivores, and initiate evolutionary changes in host plant usage by herbivores. Until recently the endemic aquatic plant Hygrophila pogonocalyx was endangered and had a restricted distribution in Taiwan. Massive restoration efforts since 1997 have led to an expansion of the plant's distribution and a novel encounter between it and an Asian butterfly, the chocolate pansy, Junonia iphita (Nymphalidae). This butterfly appears to have colonized H. pogonocalyx, switching from its original host, Strobilanthes penstemonoides var. formosana. In the present study, we aimed to investigate whether the utilization of H. pogonocalyx as a host plant has initiated a differentiation between butterflies using the novel and the original hosts. To this purpose we collected butterflies from patches of the two host plants which grow sympatrically. We tested oviposition preference for the two hosts and larval performance on them. Female adults exhibited distinct oviposition preference toward the host plant their mothers preferred. Offspring showed greater survivorship and pupal weight when fed on the host plant their mothers preferred. Male adults displayed territorial behaviors on the host plant that their mothers had preferred. Finally, the survival rate of offspring produced from cross-mating between individuals with different host plant preference was lower than that of non-hybrids. Taken together, we suggest that genetic differentiation has occurred between individuals preferring H. pogonocalyx versus S. penstemonoides as host plants via host shifting. This process was likely induced by the mass restoration of the formerly rare and endangered plant species.

  7. Butterfly Wings Are Three-Dimensional: Pupal Cuticle Focal Spots and Their Associated Structures in Junonia Butterflies.

    Science.gov (United States)

    Taira, Wataru; Otaki, Joji M

    2016-01-01

    Butterfly wing color patterns often contain eyespots, which are developmentally determined at the late larval and early pupal stages by organizing activities of focal cells that can later form eyespot foci. In the pupal stage, the focal position of a future eyespot is often marked by a focal spot, one of the pupal cuticle spots, on the pupal surface. Here, we examined the possible relationships of the pupal focal spots with the underneath pupal wing tissues and with the adult wing eyespots using Junonia butterflies. Large pupal focal spots were found in two species with large adult eyespots, J. orithya and J. almana, whereas only small pupal focal spots were found in a species with small adult eyespots, J. hedonia. The size of five pupal focal spots on a single wing was correlated with the size of the corresponding adult eyespots in J. orithya. A pupal focal spot was a three-dimensional bulge of cuticle surface, and the underside of the major pupal focal spot exhibited a hollowed cuticle in a pupal case. Cross sections of a pupal wing revealed that the cuticle layer shows a curvature at a focal spot, and a positional correlation was observed between the cuticle layer thickness and its corresponding cell layer thickness. Adult major eyespots of J. orithya and J. almana exhibited surface elevations and depressions that approximately correspond to the coloration within an eyespot. Our results suggest that a pupal focal spot is produced by the organizing activity of focal cells underneath the focal spot. Probably because the focal cell layer immediately underneath a focal spot is thicker than that of its surrounding areas, eyespots of adult butterfly wings are three-dimensionally constructed. The color-height relationship in adult eyespots might have an implication in the developmental signaling for determining the eyespot color patterns.

  8. Butterfly Wings Are Three-Dimensional: Pupal Cuticle Focal Spots and Their Associated Structures in Junonia Butterflies.

    Directory of Open Access Journals (Sweden)

    Wataru Taira

    Full Text Available Butterfly wing color patterns often contain eyespots, which are developmentally determined at the late larval and early pupal stages by organizing activities of focal cells that can later form eyespot foci. In the pupal stage, the focal position of a future eyespot is often marked by a focal spot, one of the pupal cuticle spots, on the pupal surface. Here, we examined the possible relationships of the pupal focal spots with the underneath pupal wing tissues and with the adult wing eyespots using Junonia butterflies. Large pupal focal spots were found in two species with large adult eyespots, J. orithya and J. almana, whereas only small pupal focal spots were found in a species with small adult eyespots, J. hedonia. The size of five pupal focal spots on a single wing was correlated with the size of the corresponding adult eyespots in J. orithya. A pupal focal spot was a three-dimensional bulge of cuticle surface, and the underside of the major pupal focal spot exhibited a hollowed cuticle in a pupal case. Cross sections of a pupal wing revealed that the cuticle layer shows a curvature at a focal spot, and a positional correlation was observed between the cuticle layer thickness and its corresponding cell layer thickness. Adult major eyespots of J. orithya and J. almana exhibited surface elevations and depressions that approximately correspond to the coloration within an eyespot. Our results suggest that a pupal focal spot is produced by the organizing activity of focal cells underneath the focal spot. Probably because the focal cell layer immediately underneath a focal spot is thicker than that of its surrounding areas, eyespots of adult butterfly wings are three-dimensionally constructed. The color-height relationship in adult eyespots might have an implication in the developmental signaling for determining the eyespot color patterns.

  9. Construction and Expression of Periplanete fuliginosa densovirus Recombinant Plasmid Which Contains Luciferase Gene%含荧光素酶基因的黑胸大蠊浓核病毒重组质粒的构建与表达

    Institute of Scientific and Technical Information of China (English)

    杨娟; 张珈敏; 蒋洪; 邓晓军; 胡建芳; 胡远扬

    2003-01-01

    A luciferase gene has been inserted into the recombinant plasmid PfDNV-pUC119 which contained partly deletion of genome of Periplanete fuliginosa densovirus(PfDNV.)The recombinant plasmid with luciferase gene was co-transfrected with PfDNV-pUC 119 into Periplanele fuliginosa larvae and had a high luciferase gene expression in enteron of the transfected larvae.

  10. Persistence of Penaeus stylirostris densovirus delays mortality caused by white spot syndrome virus infection in black tiger shrimp (Penaeus monodon)

    Science.gov (United States)

    2013-01-01

    Background Persistent infection of Penaeus stylirostris densovirus (PstDNV) (also called IHHNV) and its non-infectious inserts in the black tiger shrimp, Penaeus monodon (P. monodon) genome are commonly found without apparent disease. Here, we introduced the method of multiplex PCR in order to differentiate shrimp with viral inserts from ones with the infectious virus. The method allowed us to study the effect of pre-infection of IHHNV, in comparison to IHHNV inserts, on WSSV resistance in P. monodon. Results A multiplex PCR system was developed to amplify the entire IHHNV genome, ensuring the accurate diagnosis. Field samples containing IHHNV DNA templates as low as 20 pg or equivalent 150 viral copies can be detected by this method. By challenging the two groups of diagnosed shrimp with WSSV, we found that shrimp with IHHNV infection and those with viral inserts responded to WSSV differently. Considering cumulative mortality, average time to death of shrimp in IHHNV-infected group (day 14) was significantly delayed relative to that (day 10) of IHHNV-inserted group. Real-time PCR analysis of WSSV copy number indicated the lower amount of WSSV in the IHHNV-infected group than the virus-inserted group. The ratio of IHHNV: WSSV copy number in all determined IHHNV-infected samples ranged from approximately 4 to 300-fold. Conclusion The multiplex PCR assay developed herein proved optimal for convenient differentiation of shrimp specimens with real IHHNV infection and those with insert types. Diagnosed shrimp were also found to exhibit different WSSV tolerance. After exposed to WSSV, the naturally pre-infected IHHNV P. monodon were less susceptible to WSSV and, consequently, survived longer than the IHHNV-inserted shrimp. PMID:23414329

  11. Bioinformatics analysis of mosquito densovirus nostructure protein NS1%蚊浓核病毒非结构蛋白1的生物信息学分析

    Institute of Scientific and Technical Information of China (English)

    董云巧; 马文丽; 顾金保; 郑文岭

    2009-01-01

    Objective To analyze and predict the structure and function of mosquito densovirus(MDV)nostmetual proteinl(NSI).Methods Using different bioinformafics software,the EXPASY pmtparam tool,ClustalX1.83,Bioedit,MEGA3.1,ScanProsite,and Motifscan,respectively to comparatively analyze and predict the physic-chemical parameters,homology,evolutionary relation,secondary structure and main functional motifs of NS1.Results MDV NS1 protein wag a unstable hydrophilic protein and the amino acid sequence wag highly conserved which had a relatively closer evolutionary distance with infectious hypodermal and hematopoietic necrosis virus(IHHNV).MDV NS1 hag a specific domain of superfamily 3 helicage of small DNA viruses.This domain contains the NTP-binding region with a metal ion-dependent ATPase activity.A virus replication roller rolling-circle replication(RCR)initiation domain wag found near the N terminal of this protein.This protien has the biological function of single stranded incision enzyme.Conclusion The bioinformatics prediction results suggest that MDV NS1 protein phys a key role in viral replication,packaging,and the other stages of viral life.%目的 通过生物信息学分析软件对蚊浓核病毒(mosquito densovirus,MDV)菲结构蛋白1(Nostructual proteinl,NS1)的结构与功能进行分析预测.方法 应用EXPASY pmtparam tool、ClagtalX1.83、Bioedit、MEGA3.1、ScanProsite、Motifscan等分析软件和在线生物信息学分析工具对MDVNS1的理化特性、同源性、进化关系、二级结构与主要功能结构域进行分析预测.结果 MDV NSI属不稳定亲水性蛋白,氨基酸序列高度保守,与虾传染性皮下及造血组织坏死病毒(Infectious hypodermal and hematopoietic necrosis virus,IHHNV)进化距离较近;MDV NSI具有小分子DNA病毒编码的解螺旋酶超家族3(Saperfamily 3 helicage of DNA viruses)的结构域,该结构域包含有NTP-binding区域,使其具有金属离子依赖的ATPase的活性,蛋白靠近N端包

  12. Cloning and characterization of the densoviruses susceptible gene ...

    African Journals Online (AJOL)

    USER

    2010-06-21

    Jun 21, 2010 ... parvoviruses that are highly pathogenic for invertebrates and are commonly isolated from arthropod hosts (Bergoin and Tijssen, 2000). Bombyx mori ... Nid-1 gene control non-infection to BmDNV-1 (Eguchi et al., 1986) and a ...

  13. Eyespot evolution: phylogenetic insights from Junonia and related butterfly genera (Nymphalidae: Junoniini).

    Science.gov (United States)

    Kodandaramaiah, Ullasa

    2009-01-01

    Butterfly eyespots have been the focus of a number of developmental and evolutionary studies. However, a phylogenetic component has rarely been explicitly incorporated in these studies. In this study, I utilize a phylogeny to trace the evolution of eyespot number and position on the wing in a group of nymphalid butterflies, the subtribe Junoniini. These butterflies have two kinds of eyespot arrangements which I refer to as Serial and Individual. In the Serial arrangement, eyespots are placed in a series on compartments 1-6 (counting from the anterior wing margin). In the Individual arrangement, eyespots are isolated on specific compartments, ranging from 1 to 4 in number. This can be divided into four subtypes based on the number and positions of eyespots. I map the evolution of these five arrangements over a phylogeny of Junoniini reconstructed with ca. 3000 base pairs of sequence data from three genes. The results show that almost all arrangements have evolved at least twice, with multiple shifts between them by addition and deletion of eyespots. I propose a model involving genetic or developmental coupling between eyespots in specific compartments to explain these shifts. I discuss their evolution in light of existing knowledge about their development. I also discuss potential explanations for functional significance of the eyespot patterns found in the group. Differential selection for and against eyespots, both at different times over the phylogeny and in different regions, have driven the evolution of eyespot arrangements. The study throws open many questions about the adaptive significance of eyespots and the developmental underpinnings of the various arrangements.

  14. Messages from the Other Side: Parasites Receive Damage Cues from their Host Plants.

    Science.gov (United States)

    Tjiurutue, Muvari Connie; Stevenson, Philip C; Adler, Lynn S

    2016-08-01

    As sessile organisms, plants rely on their environment for cues indicating imminent herbivory. These cues can originate from tissues on the same plant or from different individuals. Since parasitic plants form vascular connections with their host, parasites have the potential to receive cues from hosts that allow them to adjust defenses against future herbivory. However, the role of plant communication between hosts and parasites for herbivore defense remains poorly investigated. Here, we examined the effects of damage to lupine hosts (Lupinus texensis) on responses of the attached hemiparasite (Castilleja indivisa), and indirectly, on a specialist herbivore of the parasite, buckeyes (Junonia coenia). Lupines produce alkaloids that act as defenses against herbivores that can be taken up by the parasite. We found that damage to lupine host plants by beet armyworm (Spodoptera exigua) significantly increased jasmonic acid (JA) levels in both the lupine host and parasite, suggesting uptake of phytohormones or priming of parasite defenses by using host cues. However, lupine host damage did not induce changes in alkaloid levels in the hosts or parasites. Interestingly, the parasite had substantially higher concentrations of JA and alkaloids compared to lupine host plants. Buckeye herbivores consumed more parasite tissue when attached to damaged compared to undamaged hosts. We hypothesize that increased JA due to lupine host damage induced higher iridoid glycosides in the parasite, which are feeding stimulants for this specialist herbivore. Our results demonstrate that damage to hosts may affect both parasites and associated herbivores, indicating cascading effects of host damage on multiple trophic levels.

  15. The relationship between total cholinesterase activity and mortality in four butterfly species

    Science.gov (United States)

    Bargar, Timothy A.

    2012-01-01

    The relationship between total cholinesterase activity (TChE) and mortality in four butterfly species (great southern white [Ascia monuste], common buckeye [Junonia coenia], painted lady [Vanessa cardui], and julia butterflies [Dryas julia]) was investigated. Acute contact toxicity studies were conducted to evaluate the response (median lethal dose [LD50] and TChE) of the four species following exposure to the organophosphate insecticide naled. The LD50 for these butterflies ranged from 2.3 to 7.6 μg/g. The average level of TChE inhibition associated with significant mortality ranged from 26 to 67%, depending on the species. The lower bounds of normal TChE activity (2 standard deviations less than the average TChE for reference butterflies) ranged from 8.4 to 12.3 μM/min/g. As a percentage of the average reference TChE activity for the respective species, the lower bounds were similar to the inhibition levels associated with significant mortality, indicating there was little difference between the dose resulting in significant TChE inhibition and that resulting in mortality.

  16. Taxonomic review of the tribe Junoniini (Lepidoptera: Nymphalidae: Nymphalinae from Myanmar

    Directory of Open Access Journals (Sweden)

    Nan Zarchi Win

    2016-09-01

    Full Text Available This paper provides keys to the genera and species for the butterfly species belonging to the tribe Junoniini (Lepidoptera: Nymphalidae from Myanmar. Species accounts include taxonomic description, synonymic lists, distributional ranges, and adult illustrations of nine species: Junonia hierta (Fabricius, Junonia orithya (Linnaeus, Junonia almana (Linnaeus, Junonia lemonias (Linnaeus, Junonia atlites (Linnaeus, Junonia iphita (Cramer, Yoma sabina (Cramer, Hypolimnas bolina (Linnaeus, and Hypolimnas misippus (Linnaeus.

  17. In vitro and in vivo host range of Anopheles gambiae densovirus (AgDNV).

    Science.gov (United States)

    Suzuki, Yasutsugu; Barik, Tapan K; Johnson, Rebecca M; Rasgon, Jason L

    2015-07-29

    AgDNV is a powerful gene transduction tool and potential biological control agent for Anopheles mosquitoes. Using a GFP reporter virus system, we investigated AgDNV host range specificity in four arthropod cell lines (derived from An. gambiae, Aedes albopictus and Drosophila melanogaster) and six mosquito species from 3 genera (An. gambiae, An. arabiensis, An. stephensi, Ae. albopictus, Ae. aegypti and Culex tarsalis). In vitro, efficient viral invasion, replication and GFP expression was only observed in MOS55 An. gambiae cells. In vivo, high levels of GFP were observed in An. gambiae mosquitoes. Intermediate levels of GFP were observed in the closely related species An. arabiensis. Low levels of GFP were observed in An. stephensi, Ae. albopictus, Ae. aegypti and Cx. tarsalis. These results suggest that AgDNV is a specific gene transduction tool for members of the An. gambiae species complex, and could be potentially developed into a biocontrol agent with minimal off-target effects.

  18. Incorporation of an introduced weed into the diet of a native butterfly: consequences for preference, performance and chemical defense.

    Science.gov (United States)

    Knerl, Angela; Bowers, M Deane

    2013-10-01

    The introduction of exotic plants, animals, and pathogens into non-native ecosystems can have profound effects on native organisms. Plantago lanceolata, narrow-leaf or ribwort plantain (Plantaginaceae), is a weed that was introduced to North America from Eurasia approximately 200 years ago and that has been incorporated into the diet of a variety of native North American herbivores. Plantain contains two iridoid glycosides, aucubin and catalpol, that can be toxic or deterrent to non-specialized herbivores or herbivores that have recently incorporated this species into their diet. Anartia jatrophae (Nymphalidae), the white peacock, feeds on plants in five families including the Plantaginaceae, and was recently observed feeding on plantain; however, the effects of feeding on this novel host plant are unknown. In this study, we performed a series of experiments to assess larval preference and performance on the introduced P. lanceolata and on a native host plant that does not contain iridoid glycosides, water hyssop, Bacopa monnieri (Plantaginaceae). We also tested whether or not white peacocks were able to sequester iridoid glycosides and compared this ability with an iridoid specialist, the buckeye, Junonia coenia (Nymphalidae). White peacocks successfully developed to the adult stage on plantain; larvae grew more slowly but pupae were heavier when compared with larvae and pupae reared on the native host plant. Larvae showed induced feeding preferences for the host plant on which they were reared. Furthermore, larvae sequestered small amounts of iridoids that were also retained in pupae and adults. These results suggest that incorporation of the introduced weed, plantain, into the diet of the white peacock may have important consequences for larval performance and preference, as well as for interactions with natural enemies.

  19. Arthromitus (Bacillus cereus) symbionts in the cockroach Blaberus giganteus: dietary influences on bacterial development and population density

    Science.gov (United States)

    Feinberg, L.; Jorgensen, J.; Haselton, A.; Pitt, A.; Rudner, R.; Margulis, L.

    1999-01-01

    The filamentous spore-forming bacterium Arthromitus, discovered in termites, millipedes, sow bugs and other soil-dwelling arthropods by Leidy (1850), is the intestinal stage of Bacillus cereus. We extend the range of Arthromitus habitats to include the hindgut of Blaberus giganteus, the large tropical American cockroach. The occurrence and morphology of the intestinal form of the bacillus were compared in individual cockroaches (n=24) placed on four different diet regimes: diurnally maintained insects fed (1) dog food, (2) soy protein only, (3)purified cellulose only, and (4) a dog food-fed group maintained in continuous darkness. Food quality exerted strong influence on population densities and developmental stages of the filamentous bacterium and on fecal pellet composition. The most dramatic rise in Arthromitus populations, defined as the spore-forming filament intestinal stage, occurred in adult cockroaches kept in the dark on a dog food diet. Limited intake of cellulose or protein alone reduced both the frequency of Arthromitus filaments and the rate of weight gain of the insects. Spores isolated from termites, sow bugs, cockroaches and moths, grown on various hard surfaces display a branching mobility and resistance to antibiotics characteristic to group I Bacilli whose members include B. cereus, B. circulans, B. alvei and B. macerans. DNA isolated from pure cultures of these bacilli taken from the guts of Blaberus giganteus (cockroach), Junonia coenia (moth), Porcellio scaber (sow bug) and Cryptotermes brevis (termite) and subjected to Southern hybridization with a 23S-5S B. subtilis ribosomal sequence probe verified that they are indistinguishable from laboratory strains of Bacillus cereus.

  20. Checklist of butterfly (Insecta: Lepidoptera fauna of Tehsil Tangi, Khyber Pakhtunkhwa, Pakistan

    Directory of Open Access Journals (Sweden)

    Farzana Khan Perveen

    2015-12-01

    Full Text Available The butterflies (Insecta: Lepidopteraare well known insects, play an important role in the ecosystem as bioindicators and pollinators. They have bright colours, remarkable shapes and supple flight. The present study was conducted to prepare the checklist of butterfly fauna of Tehsil Tangi during August, 2014 to May, 2015. A total of 506 specimens were collected belong to 3 families with 18 genera and 23 species. The collected species are the common or lemon emigrant, Catopsila ponoma Fabricius; mottled emigrant, Catopsilia pyranthe Linnaeus; clouded yellow, Colias fieldii Fabricius; common grass yellow, Eurema hecabe Linnaeus; eastern pale clouded yellow butterfly, Colias erate Esper; Indian cabbage white, Pieris canidia Sparrman; Indian little orange tip, Colotis etrida Boisduval; pioneer white or African caper white, Belonias aurota Fabricius; plain tiger, Danaua chrysippus Linnaeus; blue tiger, Tirumala liminniace Cramer; peacock pansy, Junonia almanac Linnaeus; Indian fritillary, Argyreus hyperbius Linnaeus; Indian red admiral, Venesa indica Herbst; yellow pansy, Junonia hierta Fabricius; blue pansy, Junonia orytha Linnaeus; white edged rock brown, Hipparchia parisatis Kollar; banded tree brwon, Lethe confuse Aurivillius; common castor, Ariadne merione Cramer; painted lady, Caynthia cardui Linnaeus; Himalayan sailer, Neptis mahendra Moore; common boran, Euthalia garuda Hewitson; lime butterfly, Papilio demoleus Linnaeus and great black mormon butterfly, Papilio polytes Linnaeus. It was concluded that the family Nymphalidae has the highest numbers of individuals in the present checklist. It is recommended that butterfly fauna of the study area should be conserved and their habitat should be protected.

  1. Diversity and distribution of butterflies (Insecta: Lepidoptera of district Dir lower, Khyber Pukhtoonkhwa, Pakistan

    Directory of Open Access Journals (Sweden)

    Muhammad Inayatullah Khan

    2016-03-01

    Full Text Available Butterflies are the fine-looking creatures and act as ecological indicators and pollinators. The present study is the first record of Butterfly fauna of Dir lower. Collection was carried out during March - August 2013. The specimens were collected and identified with the help of taxonomic keys and preserved specimens in National Insect Museum Islamabad. The collection of 375 specimens were preserved. Identification revealed 24 species belonging to 20 genera and 7 families. The species are Papilio polyctor Boisduval, Papilio demoleus Linnaeus, Junonia almanac Linnaeus, Pararge schakra Kollar, Junonia hierta Fabricius, Junonia orythea Linnaeus, Argyrius hyperbius Linnaeus, Hypolimnus bolina Linnaeus, Vanessa cashmiriensis Kollar, Phalantha phalantha Drury, Melitea didyma Esper, Lycaena phalaeas Linnaeus, Lybithea lipita Moore, Danius chrysippus Linnaeus, Hipparchia parasitas Kollar, Lethe rohria Fabricius, Maniola davendra Moore, Pontia daplidice Linnaeus, Belenois aurota Fabricius, Pieris brassicae Linnaeus, Colias erate Esper, Eurema hecabe Linnaeus, Colias fieldi Linnaeus and Cynthia cardui Linnaeus. The highest population was shown by Pieris brassicae followed by Danius chrysippus and Cynthia cardui. Twelve species belong to family Nymphalidae (50%, which shows the highest abundance rate. Butterfly density was the highest at Timergara. Butterfly fauna was the highest in May followed by August and lowest in March. It is concluded that pollution free environment of Dir Lower is more suitable for the survival of butterfly fauna. Large scale study is required to fully explore the butterfly fauna of the area.

  2. Spatial patterns of correlated scale size and scale color in relation to color pattern elements in butterfly wings.

    Science.gov (United States)

    Iwata, Masaki; Otaki, Joji M

    2016-02-01

    Complex butterfly wing color patterns are coordinated throughout a wing by unknown mechanisms that provide undifferentiated immature scale cells with positional information for scale color. Because there is a reasonable level of correspondence between the color pattern element and scale size at least in Junonia orithya and Junonia oenone, a single morphogenic signal may contain positional information for both color and size. However, this color-size relationship has not been demonstrated in other species of the family Nymphalidae. Here, we investigated the distribution patterns of scale size in relation to color pattern elements on the hindwings of the peacock pansy butterfly Junonia almana, together with other nymphalid butterflies, Vanessa indica and Danaus chrysippus. In these species, we observed a general decrease in scale size from the basal to the distal areas, although the size gradient was small in D. chrysippus. Scales of dark color in color pattern elements, including eyespot black rings, parafocal elements, and submarginal bands, were larger than those of their surroundings. Within an eyespot, the largest scales were found at the focal white area, although there were exceptional cases. Similarly, ectopic eyespots that were induced by physical damage on the J. almana background area had larger scales than in the surrounding area. These results are consistent with the previous finding that scale color and size coordinate to form color pattern elements. We propose a ploidy hypothesis to explain the color-size relationship in which the putative morphogenic signal induces the polyploidization (genome amplification) of immature scale cells and that the degrees of ploidy (gene dosage) determine scale color and scale size simultaneously in butterfly wings.

  3. Ochre star mortality during the 2014 wasting disease epizootic: role of population size structure and temperature

    OpenAIRE

    2016-01-01

    Over 20 species of asteroids were devastated by a sea star wasting disease (SSWD) epizootic, linked to a densovirus, from Mexico to Alaska in 2013 and 2014. For Pisaster ochraceus from the San Juan Islands, South Puget Sound and Washington outer coast, time-series monitoring showed rapid disease spread, high mortality rates in 2014, and continuing levels of wasting in the survivors in 2015. Peak prevalence of disease at 16 sites ranged to 100%, with an overall mean of 61%. Analysis of longitu...

  4. Butterfly Diversity of Indian Institute of Forest Management, Bhopal, Madhya Pradesh, India

    Directory of Open Access Journals (Sweden)

    Sprih Harsh

    2014-01-01

    Full Text Available A study to find out the diversity of butterflies at the Indian Institute of Forest Management (IIFM, Bhopal, was carried out over a period of six months from October 2013 to March 2014. A total of 55 butterfly species belonging to 5 families, namely, Hesperiidae (7 species, Papilionidae (4 species, Pieridae (10 species, Lycaenidae (13 species, and Nymphalidae (21 species, were recorded (with photographic record during the study from three different habitats of campus: open scrub, dry deciduous, and urbanized habitat. Shannon diversity indices and Pielou’s evenness index were calculated for all the habitats. Shannon index was found to be highest for open scrub (3.76. Out of 54 species, Eurema brigitta was the most dominant species followed by Eurema hecabe, Junonia lemonias, and Phalanta phalantha. Dominance of these species can be explained by the presence of their larval and host plants in the campus.

  5. Avian pox in white-tailed laurel-pigeons from the Canary Islands.

    Science.gov (United States)

    Manuel Medina, Félix; Adolfo Ramírez, Gustavo; Hernández, Antonio

    2004-04-01

    Two diseased young white-tailed laurel-pigeons (Columba junoniae), an endemic and endangered species of the Canary Islands (Spain), were found in La Palma. They were very depressed and had severe cutaneous yellowish nodular lesions in feathered and unfeathered areas on the bodies of both birds. Necropsy and histopathologic analyses were conducted. The presence of epidermal hypertrophy and hyperplasia in cutaneous lesions, as well as several acidophilic intracytoplasmic inclusions in affected epithelial cells (Bollinger bodies), confirmed avian poxvirus infection. This is the first report of avian pox in whitetailed laurel-pigeons or in any other free-ranging bird in the Canaries, and it might indicate that other threatened birds of the Canarian Archipelago may be affected by this viral disease.

  6. The structure and host entry of an invertebrate parvovirus.

    Science.gov (United States)

    Meng, Geng; Zhang, Xinzheng; Plevka, Pavel; Yu, Qian; Tijssen, Peter; Rossmann, Michael G

    2013-12-01

    The 3.5-Å resolution X-ray crystal structure of mature cricket parvovirus (Acheta domesticus densovirus [AdDNV]) has been determined. Structural comparisons show that vertebrate and invertebrate parvoviruses have evolved independently, although there are common structural features among all parvovirus capsid proteins. It was shown that raising the temperature of the AdDNV particles caused a loss of their genomes. The structure of these emptied particles was determined by cryo-electron microscopy to 5.5-Å resolution, and the capsid structure was found to be the same as that for the full, mature virus except for the absence of the three ordered nucleotides observed in the crystal structure. The viral protein 1 (VP1) amino termini could be externalized without significant damage to the capsid. In vitro, this externalization of the VP1 amino termini is accompanied by the release of the viral genome.

  7. History and current status of development and use of viral insecticides in China.

    Science.gov (United States)

    Sun, Xiulian

    2015-01-20

    The use of insect viruses as biological control agents started in the early 1960s in China. To date, more than 32 viruses have been used to control insect pests in agriculture, forestry, pastures, and domestic gardens in China. In 2014, 57 products from 11 viruses were authorized as commercial viral insecticides by the Ministry of Agriculture of China. Approximately 1600 tons of viral insecticidal formulations have been produced annually in recent years, accounting for about 0.2% of the total insecticide output of China. The development and use of Helicoverpa armigera nucleopolyhedrovirus, Mamestra brassicae nucleopolyhedrovirus, Spodoptera litura nucleopolyhedrovirus, and Periplaneta fuliginosa densovirus are discussed as case studies. Additionally, some baculoviruses have been genetically modified to improve their killing rate, infectivity, and ultraviolet resistance. In this context, the biosafety assessment of a genetically modified Helicoverpa armigera nucleopolyhedrovirus is discussed.

  8. History and Current Status of Development and Use of Viral Insecticides in China

    Directory of Open Access Journals (Sweden)

    Xiulian Sun

    2015-01-01

    Full Text Available The use of insect viruses as biological control agents started in the early 1960s in China. To date, more than 32 viruses have been used to control insect pests in agriculture, forestry, pastures, and domestic gardens in China. In 2014, 57 products from 11 viruses were authorized as commercial viral insecticides by the Ministry of Agriculture of China. Approximately 1600 tons of viral insecticidal formulations have been produced annually in recent years, accounting for about 0.2% of the total insecticide output of China. The development and use of Helicoverpa armigera nucleopolyhedrovirus, Mamestra brassicae nucleopolyhedrovirus, Spodoptera litura nucleopolyhedrovirus, and Periplaneta fuliginosa densovirus are discussed as case studies. Additionally, some baculoviruses have been genetically modified to improve their killing rate, infectivity, and ultraviolet resistance. In this context, the biosafety assessment of a genetically modified Helicoverpa armigera nucleopolyhedrovirus is discussed.

  9. Discovery of a novel circular DNA virus in the Forbes sea star, Asterias forbesi.

    Science.gov (United States)

    Fahsbender, Elizabeth; Hewson, Ian; Rosario, Karyna; Tuttle, Allison D; Varsani, Arvind; Breitbart, Mya

    2015-09-01

    A single-stranded DNA (ssDNA) virus, Asterias forbesi-associated circular virus (AfaCV), was discovered in a Forbes sea star displaying symptoms of sea star wasting disease (SSWD). The AfaCV genome organization is typical of circular Rep-encoding ssDNA (CRESS-DNA) viruses and is similar to that of members of the family Circoviridae. PCR-based surveys indicate that AfaCV is not clearly associated with SSWD, whereas the sea star-associated densovirus (SSaDV), recently implicated in SSWD in the Pacific, was prevalent in symptomatic specimens. AfaCV represents the first CRESS-DNA virus detected in echinoderms, adding to the growing diversity of these viruses recently recovered from invertebrates.

  10. [Do live fences help conserve butterfly diversity in agricultural landscapes?].

    Science.gov (United States)

    Tobar L, Diego Enrique; Ibrahim, Muhammad

    2010-03-01

    In Central America, natural forests have been transformed into agriculture production areas, generating forest fragmentation, desertification, erosion and loss of biodiversity, among other concerns. Different tree cover compositions are kept on these agricultural landscapes, including scattered trees in pastures, live fences, fragments of secondary forests, and riparian forests. These can help in biodiversity conservation because they generate shelter, feeding and reproduction areas, among others. We studied the composition, richness and abundance of diurnal butterflies on two types of live fences in a landscape where pastures are predominant in Costa Rica's Central Pacific Region. Transects (120 x 5 m) were observed for an hour (two days/habitat) in five multi-strata fences (with several plant species, strata and canopy width) and five simple fences (with smaller and pruned trees). A total of 2,782 butterflies were observed (75 species). The most abundant species were Anartia fatima, Eurema daria, Eurema nise, Hermeuptychia hermes, Junonia evarete and Phoebis philea. Multi-strata fences had more species and can help maintain 56% of the total species observed in secondary and riparian forests. This type of live fence can play an important role in butterfly conservation in livestock areas, and its benefits are influenced by the manner in which farmers manage their land.

  11. Molecular Phylogenetic Analysis of the Main Lineages of Nymphalinae (Nymphalidae: Lepidoptera) Based on the Partial Mitochondrial COI Gene

    Institute of Scientific and Technical Information of China (English)

    ZHANG Min; CAO Tian-wen; ZHONG Yang; REN Zhu-mei; GUO Ya-ping; MA En-bo

    2008-01-01

    The phylogenetic relationships of the subfamily Nymphalinae (sensu Chou 1994) were analyzed based on 1488bp of mtDNA cytochrome oxidase subunit I (COI) gene sequence data obtained from 24 individuals, along with those of eight species obtained from GenBank. The base compositions of this COI fragment varied among the individuals as follows: T 39.9%, C 14.6%, A 32.2%, and G 13.4%, with a strong AT bias (72.1%), as usually found in insect mitochondrial genomes. The A+T contents of the third, second, and first codon positions of the COI fragments in this study was 92.4, 62.2, and 61.4%, respectively. The phylogenetic trees were reconstructed by neighbor-joining (NJ), maximum likelihood (ML), and Bayesian methods by using Byblia anvatara as outgroup. Phylogenetic analyses based on the COI gene sequence data created very similar topologies, which were producing trees with two main clades A and B, and five subclades. The data indicated that the tribes Nymphalini and Hypolimni (sensu Chou 1994) are not monophyletic groups, and the genus Junonia should be removed from Nymphalini to Hypolimni (=Junoniini). On the basis of the data, the Symbrenthia and Araschnia had a relative distant relationship with the rest of Nymphalini. The relationships of species in the Nymphalini were confirmed via the NJ, ML, and Bayesian methods, namely ((((Nymphalis+Kaniska)+Polygonia)+Aglais)+Vanessa)+(Symbrenthia+Araschnia). This investigation provides a little novel information for Chinese researches of butterflies.

  12. Butterfly Diversity from Farmlands of Central Uganda

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    M. B. Théodore Munyuli

    2012-01-01

    Full Text Available The aim of this study was to collect information about the diversity of butterfly communities in the mixed coffee-banana mosaic (seminatural, agricultural landscapes of rural central Uganda. Data were collected for one year (2006 using fruit-bait traps, line transect walk-and-counts, and hand nets. A total of 56,315 individuals belonging to 331 species, 95 genera, and 6 families were sampled. The most abundant species was Bicyclus safitza (14.5% followed by Acraea acerata (6.3%, Catopsilia florella (6.5% and Junonia sophia (6.1%. Significant differences in abundance, species richness, and diversity of butterflies occurred between the 26 study sites. Farmland butterflies visited a variety of habitats within and around sites, but important habitats included woodlands, fallows, hedgerows, swampy habitats, abandoned gardens, and home gardens. The highest diversity and abundance of butterflies occurred in sites that contained forest remnants. Thus, forest reserves in the surrounding of fields increased the conservation values of coffee-banana agroforestry systems for butterflies. Their protection from degradation should be a priority for policy makers since they support a species-rich community of butterflies pollinating cultivated plants. Farmers are encouraged to protect and increase on-farm areas covered by complex traditional agroforests, linear, and nonlinear seminatural habitats to provide sufficient breeding sites and nectar resources for butterflies.

  13. ¿Las cercas vivas ayudan a la conservación de la diversidad de mariposas en paisajes agropecuarios?

    Directory of Open Access Journals (Sweden)

    Diego Enrique Tobar L

    2010-03-01

    Full Text Available En Centro América, los paisajes agropecuarios mantienen diferentes formas de cobertura arbórea como árboles dispersos, cercas vivas, fragmentos de bosque, bosques ribereños, que pueden generar hábitats apropiados para la conservación de la biodiversidad. El presente estudio caracterizó la composición, riqueza y abundancia de mariposas en dos tipos de cercas vivas presentes en un paisaje dominado por pasturas en el Pacifico Central de Costa Rica. Se seleccionaron un total de cinco cercas vivas por cada tipo de cerca (simples y multiestrato, donde se establecieron franjas de 120x5m que fueron recorridos por espacio de una hora durante dos días/hábitat. Se registró un total de 2 782 individuos, pertenecientes a 75 especies de mariposas. Las especies más abundantes fueron: Anartia fatima, Eurema daría, E. nise, Hermeuptychia hermes, Junonia evarete y Phoebis philea. Las cercas vivas multiestrato presentaron una mayor riqueza y abundancia de especies de mariposas que las cercas vivas simples. Las cercas vivas multiestrato pueden ayudar a mantener el 56% de las especies encontradas en los bosques secundarios y ribereños. Este tipo de cerca viva pueden jugar un papel importante para la conservación de mariposas en áreas de producción pecuaria, y su beneficio está influenciado por el tipo manejo que realizan los productores.

  14. Ochre star mortality during the 2014 wasting disease epizootic: role of population size structure and temperature

    Science.gov (United States)

    Eisenlord, Morgan E.; Yoshioka, Reyn M.; Elliott, Joel; Maynard, Jeffrey; Fradkin, Steven; Turner, Margaret; Pyne, Katie; Rivlin, Natalie; van Hooidonk, Ruben; Harvell, C. Drew

    2016-01-01

    Over 20 species of asteroids were devastated by a sea star wasting disease (SSWD) epizootic, linked to a densovirus, from Mexico to Alaska in 2013 and 2014. For Pisaster ochraceus from the San Juan Islands, South Puget Sound and Washington outer coast, time-series monitoring showed rapid disease spread, high mortality rates in 2014, and continuing levels of wasting in the survivors in 2015. Peak prevalence of disease at 16 sites ranged to 100%, with an overall mean of 61%. Analysis of longitudinal data showed disease risk was correlated with both size and temperature and resulted in shifts in population size structure; adult populations fell to one quarter of pre-outbreak abundances. In laboratory experiments, time between development of disease signs and death was influenced by temperature in adults but not juveniles and adult mortality was 18% higher in the 19°C treatment compared to the lower temperature treatments. While larger ochre stars developed disease signs sooner than juveniles, diseased juveniles died more quickly than diseased adults. Unusual 2–3°C warm temperature anomalies were coincident with the summer 2014 mortalities. We suggest these warm waters could have increased the disease progression and mortality rates of SSWD in Washington State. PMID:26880844

  15. [Investigation on mosquitoes and mosquito-borne viruses in Dehong prefecture, Yunnan province, 2007 and 2010].

    Science.gov (United States)

    Feng, Yun; Zhang, Hailin; Fu, Shihong; Yang, Weihong; Zhang, Yuzhen; Wang, Piyu; Yang, Jie; Liu, Yonghua; Dong, Chaoliang; Li, Shi; Zhang, Baosen; Yin, Zhengliu; Dong, Xingqi; Wang, Huanyu; Liang, Guodong

    2014-05-01

    To investigate the distribution patterns of mosquito and mosquito-borne viruses in Dehong prefecture, Yunnan province, China. Mosquito samples were collected using the mosquito traps from five counties of Dehong prefecture on July, 2007 and 2010. Mosquito were cell cultured for viral isolation, and positive isolates were identified using RT-PCR and sequence analysis. A total of 43 634 mosquito comprised of 29 species representing six genera were collected. Culex tritaeniorhynchus and Anopheles sinensis comprised 78.69% and 14.77% of the total. Six strains of viruses were isolated from the mosquito pools. RT-PCR and phylogenetic analysis revealed three strains from Cx. tritaeniorhynchus, identified as genotype I Japanese encephalitis virus (JEV). One strain was identified from Cx. tritaeniorhynchus, as Getah virus (GETV). Two strains isolated from Cx. tritaeniorhynchus and Anopheles vagus were identified as Culex pipiens pallens Densovirus (CppDNV). Cx. tritaeniorhynchus had been the major species of mosquito and mainly transmitting vector of mosquito-borne viruses in Dehong prefecture. Genotype I JEV, GETV and CppDNV were the vectors causing transmission of mosquito-borne diseases in this area. Data from phylogenetic analysis showed that these newly discovered isolates seemed to have had close relationship with those viruses previously circulating in Yunnan and other provinces of China.

  16. Selection of reference genes for analysis of stress-responsive genes after challenge with viruses and temperature changes in the silkworm Bombyx mori.

    Science.gov (United States)

    Guo, Huizhen; Jiang, Liang; Xia, Qingyou

    2016-04-01

    Viruses and high temperature (HT) are the primary threats to silkworms. Changes in the expression of stress-response genes can be measured using quantitative polymerase chain reaction (qPCR) after exposure to viruses or HT. However, appropriate reference genes (RGs) for qPCR data normalization have not been established in this organism. In this study, we summarized the RGs used in the previous silkworm studies after infection with Bombyx mori nucleopolyhedrovirus (BmNPV), B. mori cytoplasmic polyhedrosis virus (BmCPV), or B. mori densovirus (BmDNV) or after HT treatment. The expression levels of these RGs were extracted from silkworm transcriptome data to screen for candidate RGs that were unaffected by the experimental conditions. Actin-1 (A1), actin-3 (A3), glyceraldehyde-3-phosphate dehydrogenase (GAPDH), and translation initiation factor 4a (TIF-4A) were selected for further qPCR verification. The results of RNA-seq and qPCR showed that GAPDH and TIF-4A were suitable RGs after BmNPV challenge or HT stress, whereas TIF-4A was an appropriate RG for BmCPV or BmDNV-Z challenge in silkworms. These results suggested that TIF-4A may be the most appropriate RG for gene expression analysis after challenge with viruses or HT in silkworms.

  17. AaeDV编码的miRNA样分子的分析与鉴定%Identification of MicroRNA-like Viral Small RNAs from AaeDV

    Institute of Scientific and Technical Information of China (English)

    王衍海; 吴江; 顾金保

    2015-01-01

    一些双链DNA病毒和RNA病毒可以在宿主细胞内编码自身的miRNAs分子,其在病毒感染宿主细胞与病毒复制过程中发挥重要的作用,为研究单链DNA病毒是否可编码miRNAs,我们利用VMir、miRNAFold和MaturePred软件,通过生物信息学方法预测了一株单链DNA病毒-埃及伊蚊浓核病毒(Aedes aegypti densovirus,AaeDV)编码的miRNAs分子的颈环结构前体与成熟体,并通过Northern blot与stem-loop RT-PCR方法进行了验证,结果在预测的4个茎环结构前体中,通过stem-loop RT-PCR方法验证到一个miRNA样分子AaeDVMD.该结果证实了单链DNA病毒可编码miRNAs,为进一步研究蚊浓核病毒与宿主蚊类之间的相互作用提供了新的研究方向.

  18. A viral over-expression system for the major malaria mosquito Anopheles gambiae.

    Science.gov (United States)

    Suzuki, Yasutsugu; Niu, Guodong; Hughes, Grant L; Rasgon, Jason L

    2014-05-30

    Understanding pathogen/mosquito interactions is essential for developing novel strategies to control mosquito-borne diseases. Technical advances in reverse-genetics, such as RNA interference (RNAi), have facilitated elucidation of components of the mosquito immune system that are antagonistic to pathogen development, and host proteins essential for parasite development. Forward genetic approaches, however, are limited to generation of transgenic insects, and while powerful, mosquito transgenesis is a resource- and time-intensive technique that is not broadly available to most laboratories. The ability to easily "over-express" genes would enhance molecular studies in vector biology and expedite elucidation of pathogen-refractory genes without the need to make transgenic insects. We developed and characterized an efficient Anopheles gambiae densovirus (AgDNV) over-expression system for the major malaria vector Anopheles gambiae. High-levels of gene expression were detected at 3 days post-infection and increased over time, suggesting this is an effective system for gene induction. Strong expression was observed in the fat body and ovaries. We validated multiple short promoters for gene induction studies. Finally, we developed a polycistronic system to simultaneously express multiple genes of interest. This AgDNV-based toolset allows for consistent transduction of genes of interest and will be a powerful molecular tool for research in Anopheles gambiae mosquitoes.

  19. Virus and calcium: an unexpected tandem to optimize insecticide efficacy.

    Science.gov (United States)

    Apaire-Marchais, Véronique; Ogliastro, Mylène; Chandre, Fabrice; Pennetier, Cédric; Raymond, Valérie; Lapied, Bruno

    2016-04-01

    The effective control of insect pests is based on the rational use of the most efficient and safe insecticide treatments. To increase the effects of classical insecticides and to avoid the ability of certain pest insects to develop resistance, it is essential to propose novel strategies. Previous studies have shown that calcium-dependent phosphorylation/dephosphorylation is now considered as a new cellular mechanism for increasing the target sensitivity to insecticides. Because it is known that virus entry is correlated with intracellular calcium concentration rise, this report attempts to present the most important data relevant to the feasibility of combining an insect virus such as baculovirus or densovirus with an insecticide. In this case, the insect virus is not used as a bioinsecticide but acts as a synergistic agent able to trigger calcium rise and to activate calcium-dependent intracellular signalling pathways involved in the increase of the membrane receptors and/or ion channels sensitivity to insecticides. This virus-insecticide mixture represents a promising alternative to optimize the efficacy of insecticides against insect pests while reducing the doses. © 2016 Society for Applied Microbiology and John Wiley & Sons Ltd.

  20. ¿Las cercas vivas ayudan a la conservación de la diversidad de mariposas en paisajes agropecuarios?

    Directory of Open Access Journals (Sweden)

    Diego Enrique Tobar L

    2010-03-01

    Full Text Available En Centro América, los paisajes agropecuarios mantienen diferentes formas de cobertura arbórea como árboles dispersos, cercas vivas, fragmentos de bosque, bosques ribereños, que pueden generar hábitats apropiados para la conservación de la biodiversidad. El presente estudio caracterizó la composición, riqueza y abundancia de mariposas en dos tipos de cercas vivas presentes en un paisaje dominado por pasturas en el Pacifico Central de Costa Rica. Se seleccionaron un total de cinco cercas vivas por cada tipo de cerca (simples y multiestrato, donde se establecieron franjas de 120x5m que fueron recorridos por espacio de una hora durante dos días/hábitat. Se registró un total de 2 782 individuos, pertenecientes a 75 especies de mariposas. Las especies más abundantes fueron: Anartia fatima, Eurema daría, E. nise, Hermeuptychia hermes, Junonia evarete y Phoebis philea. Las cercas vivas multiestrato presentaron una mayor riqueza y abundancia de especies de mariposas que las cercas vivas simples. Las cercas vivas multiestrato pueden ayudar a mantener el 56% de las especies encontradas en los bosques secundarios y ribereños. Este tipo de cerca viva pueden jugar un papel importante para la conservación de mariposas en áreas de producción pecuaria, y su beneficio está influenciado por el tipo manejo que realizan los productores.Do live fences help conserve butterfly diversity in agricultural landscapes? In Central America, natural forests have been transformed into agriculture production areas, generating forest fragmentation, desertification, erosion and loss of biodiversity, among other concerns. Different tree cover compositions are kept on these agricultural landscapes, including scattered trees in pastures, live fences, fragments of secondary forests, and riparian forests. These can help in biodiversity conservation because they generate shelter, feeding and reproduction areas, among others. We studied the composition, richness and

  1. Parasites of Columba livia (Aves: Columbiformes in Tenerife (Canary Islands and their role in the conservation biology of the Laurel pigeons

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    Foronda P.

    2004-09-01

    Full Text Available The prevalence and intensity of the parasites from 50 wild doves (Columba livia from the city of Santa Cruz de Tenerife, in the island of Tenerife (Canary Archipelago, were studied. The following ectoparasites were found in apparently healthy pigeons (prevalences are shown in percentage (% and mean intensities with their standard deviations: the acari Dermanyssus gallinae (De Geer, 1778 (6 241 .0 ± 138.9 and Tinaminyssus melloi Fain, 1962 (10 %, 218.3 ± 117.3; the louses, Columbicola columbae Linnaeus, 1758 (100 %, 111.4 ± 76.8 and Campanulotes bidentatus Scopoli, 1763 (94 %, 48.4 ± 26.6; and the pigeon fly, Pseudolynchia canariensis Macquart, 1839 (36 %, 6.2 ± 1.6. The endoparasites we found, were: a haemoprotozoan species, Haemoproteus columbae Kruze, 1890 (82 %, 14.8 ± 10.3 per 1000; coccidian oocysts, Eimeria sp. (50 %, 0.2 x 103 ± 1.7 x 103 per gr; a cestode species Raillietina micracantha (Fuhrmann, 1909 López Neyra, 1947 (44 %, 12.3 ± 9.4; and four nematode species, Tetrameres (Tetrameres fissispina (Diesing, 1861 Travassos, 1915 (4 %, 99.5 ± 34,1, Synhimantus (Dispharynx spiralis (Molin, 1858 (8 %, 46. 8 ± 11.6, Ascaridia columbae (Gmelin, 1790 Travassos, 1913 (40 %, 8.4 ± 8.8 and Aonchotheca sp. (18 %, 6.0 ± 3.1. Several species detected in our study can be pathogens for C. bollii and C. junoniae, which are endemic pigeons of the Canary Islands, considered endangered species. Parasites (ectoparasites, protozoa and helminths of C. livia found in Tenerife and others from wild and farm birds in the island were considered as healthy controls.

  2. Spontaneous long-range calcium waves in developing butterfly wings.

    Science.gov (United States)

    Ohno, Yoshikazu; Otaki, Joji M

    2015-03-25

    Butterfly wing color patterns emerge as the result of a regular arrangement of scales produced by epithelial scale cells at the pupal stage. These color patterns and scale arrangements are coordinated throughout the wing. However, the mechanism by which the development of scale cells is controlled across the entire wing remains elusive. In the present study, we used pupal wings of the blue pansy butterfly, Junonia orithya, which has distinct eyespots, to examine the possible involvement of Ca(2+) waves in wing development. Here, we demonstrate that the developing pupal wing tissue of the blue pansy butterfly displayed spontaneous low-frequency Ca(2+) waves in vivo that propagated slowly over long distances. Some waves appeared to be released from the immediate peripheries of the prospective eyespot and discal spot, though it was often difficult to identify the specific origins of these waves. Physical damage, which is known to induce ectopic eyespots, led to the radiation of Ca(2+) waves from the immediate periphery of the damaged site. Thapsigargin, which is a specific inhibitor of Ca(2+)-ATPases in the endoplasmic reticulum, induced an acute increase in cytoplasmic Ca(2+) levels and halted the spontaneous Ca(2+) waves. Additionally, thapsigargin-treated wings showed incomplete scale development as well as other scale and color pattern abnormalities. We identified a novel form of Ca(2+) waves, spontaneous low-frequency slow waves, which travel over exceptionally long distances. Our results suggest that spontaneous Ca(2+) waves play a critical role in the coordinated development of scale arrangements and possibly in color pattern formation in butterflies.

  3. On the origins of sexual dimorphism in butterflies.

    Science.gov (United States)

    Oliver, Jeffrey C; Monteiro, Antónia

    2011-07-01

    The processes governing the evolution of sexual dimorphism provided a foundation for sexual selection theory. Two alternative processes, originally proposed by Darwin and Wallace, differ primarily in the timing of events creating the dimorphism. In the process advocated by Darwin, a novel ornament arises in a single sex, with no temporal separation in the origin and sex-limitation of the novel trait. By contrast, Wallace proposed a process where novel ornaments appear simultaneously in both sexes, but are then converted into sex-limited expression by natural selection acting against showy coloration in one sex. Here, we investigate these alternative modes of sexual dimorphism evolution in a phylogenetic framework and demonstrate that both processes contribute to dimorphic wing patterns in the butterfly genera Bicyclus and Junonia. In some lineages, eyespots and bands arise in a single sex, whereas in other lineages they appear in both sexes but are then lost in one of the sexes. In addition, lineages displaying sexual dimorphism were more likely to become sexually monomorphic than they were to remain dimorphic. This derived monomorphism was either owing to a loss of the ornament ('drab monomorphism') or owing to a gain of the same ornament by the opposite sex ('mutual ornamentation'). Our results demonstrate the necessity of a plurality in theories explaining the evolution of sexual dimorphism within and across taxa. The origins and evolutionary fate of sexual dimorphism are probably influenced by underlying genetic architecture responsible for sex-limited expression and the degree of intralocus sexual conflict. Future comparative and developmental work on sexual dimorphism within and among taxa will provide a better understanding of the biases and constraints governing the evolution of animal sexual dimorphism.

  4. Insect Diversity of the Muni-Pomadze Ramsar Site: An Important Site for Biodiversity Conservation in Ghana

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    Rosina Kyerematen

    2014-01-01

    Full Text Available An inventory of species diversity of insects of the Muni-Pomadze Ramsar site, with special reference to species of conservation concern, was carried out as part of an evaluation of changes in the ecological character of the site, twenty years after designation. Samples were taken from two protected areas within the Ramsar site, in the wet (July, dry (January, and intermediate (June seasons. Community diversity was characterized in terms of number of species accumulated, species richness, Shannon-Weiner indices of diversity, Pielou’s evenness, and Bray-Curtis similarity. A total of 134 families from 19 insect orders were recorded during the entire study period. Yenku Block A recorded the highest species richness (98 and the highest diversity index (14.97, corroborated by the highest Margalef index of 3.82 with a relatively even distribution of species (0.834 during the intermediate season, and recorded the lowest diversity (6.957 and species richness (41 during the dry season. On the whole, the Muni-Pomadzi Ramsar site showed a high diversity of insect species. The presence of species such as Junonia oenone and Papilio demodocus which are specialized in degraded habitats at Yenku Block A in large numbers is a clear indication of degradation of the forest, but the presence of forest species such as Salamis anacardii and Euphaedra crokeri is an indication that some parts of this reserve are still in good shape. A comparison of the butterfly species recorded with findings in a 1997 survey showed a marked increase in numbers from 75 to 130; this may be attributed to the habitat changes that have taken place at the site offering more diverse habitat types.

  5. Parasites of Columba livia (Aves: Columbiformes) in Tenerife (Canary Islands) and their role in the conservation biology of the laurel pigeons.

    Science.gov (United States)

    Foronda, P; Valladares, B; Rivera-Medina, J A; Figueruelo, E; Abreu, N; Casanova, J C

    2004-09-01

    The prevalence and intensity of the parasites from 50 wild doves (Columba livia) from the city of Santa Cruz de Tenerife, in the island of Tenerife (Canary Archipelago), were studied. The following ectoparasites were found in apparently healthy pigeons (prevalences are shown in percentage (%) and mean intensities with their standard deviations): the acari Dermanyssus gallinae (De Geer, 1778) (6, 241.0 +/- 138.9) and Tinaminyssus melloi Fain, 1962 (10%, 218.3 +/- 117.3); the louses, Columbicola columbae Linnaeus, 1758 (100%, 111.4 +/- 76.8) and Campanulotes bidentatus Scopoli, 1763 (94%, 48.4 +/- 26.6); and the pigeon fly, Pseudolynchia canariensis Macquart, 1839 (36%, 6.2 +/- 1.6). The endoparasites we found were: a haemoprotozoan species, Haemoproteus columbae Kruze, 1890 (82%, 14.8 +/- 10.3 per 1000); coccidian oocysts, Eimeria sp. (50%, 0.2 x 10(3) +/- 1.7 x 10(3) per gr); a cestode species Raillietina micracantha (Fuhrmann, 1909) López Neyra, 1947 (44%, 12.3 +/- 9.4); and four nematode species, Tetrameres (Tetrameres) fissispina (Diesing, 1861) Travassos, 1915 (4%, 99.5 +/- 34.1), Synhimantus (Dispharynx) spiralis (Molin, 1858) (8%, 46.8 +/- 11.6), Ascaridia columbae (Gmelin, 1790) Travassos, 1913 (40%, 8.4 +/- 8.8) and Aonchotheca sp. (18%, 6.0 +/- 3.1). Several species detected in our study can be pathogens for C. bollii and C. junoniae, which are endemic pigeons of the Canary Islands, considered endangered species. Parasites (ectoparasites, protozoa and helminths) of C. livia found in Tenerife and others from wild and farm birds in the island were considered as healthy controls.

  6. A Single-Wing Removal Method to Assess Correspondence Between Gene Expression and Phenotype in Butterflies: The Case of Distal-less.

    Science.gov (United States)

    Adhikari, Kiran; Otaki, Joji M

    2016-02-01

    It is often desirable but difficult to retrieve information on the mature phenotype of an immature tissue sample that has been subjected to gene expression analysis. This problem cannot be ignored when individual variation within a species is large. To circumvent this problem in the butterfly wing system, we developed a new surgical method for removing a single forewing from a pupa using Junonia orithya; the operated pupa was left to develop to an adult without eclosion. The removed right forewing was subjected to gene expression analysis, whereas the non-removed left forewing was examined for color patterns. As a test case, we focused on Distal-less (Dll), which likely plays an active role in inducing elemental patterns, including eyespots. The Dll expression level in forewings was paired with eyespot size data from the same individual. One third of the operated pupae survived and developed wing color patterns. Dll expression levels were significantly higher in males than in females, although male eyespots were smaller in size than female eyespots. Eyespot size data showed weak but significant correlations with the Dll expression level in females. These results demonstrate that a single-wing removal method was successfully applied to the butterfly wing system and suggest the weak and non-exclusive contribution of Dll to eyespot size determination in this butterfly. Our novel methodology for establishing correspondence between gene expression and phenotype can be applied to other candidate genes for color pattern development in butterflies. Conceptually similar methods may also be applicable in other developmental systems.

  7. Persistent, triple-virus co-infections in mosquito cells

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    Malasit Prida

    2010-01-01

    Full Text Available Abstract Background It is known that insects and crustaceans can carry simultaneous, active infections of two or more viruses without showing signs of disease, but it was not clear whether co-infecting viruses occupied the same cells or different cells in common target tissues. Our previous work showed that successive challenge of mosquito cell cultures followed by serial, split-passage resulted in stabilized cultures with 100% of the cells co-infected with Dengue virus (DEN and an insect parvovirus (densovirus (DNV. By addition of Japanese encephalitis virus (JE, we tested our hypothesis that stable, persistent, triple-virus co-infections could be obtained by the same process. Results Using immunocytochemistry by confocal microscopy, we found that JE super-challenge of cells dually infected with DEN and DNV resulted in stable cultures without signs of cytopathology, and with 99% of the cells producing antigens of the 3 viruses. Location of antigens for all 3 viruses in the triple co-infections was dominant in the cell nuclei. Except for DNV, this differed from the distribution in cells persistently infected with the individual viruses or co-infected with DNV and DEN. The dependence of viral antigen distribution on single infection or co-infection status suggested that host cells underwent an adaptive process to accommodate 2 or more viruses. Conclusions Individual mosquito cells can accommodate at least 3 viruses simultaneously in an adaptive manner. The phenomenon provides an opportunity for genetic exchange between diverse viruses and it may have important medical and veterinary implications for arboviruses.

  8. Citizen Science: The First Peninsular Malaysia Butterfly Count.

    Science.gov (United States)

    Wilson, John-James; Jisming-See, Shi-Wei; Brandon-Mong, Guo-Jie; Lim, Aik-Hean; Lim, Voon-Ching; Lee, Ping-Shin; Sing, Kong-Wah

    2015-01-01

    Over the past 50 years, Southeast Asia has suffered the greatest losses of biodiversity of any tropical region in the world. Malaysia is a biodiversity hotspot in the heart of Southeast Asia with roughly the same number of mammal species, three times the number of butterfly species, but only 4% of the land area of Australia. Consequently, in Malaysia, there is an urgent need for biodiversity monitoring and also public engagement with wildlife to raise awareness of biodiversity loss. Citizen science is "on the rise" globally and can make valuable contributions to long-term biodiversity monitoring, but perhaps more importantly, involving the general public in science projects can raise public awareness and promote engagement. Butterflies are often the focus of citizen science projects due to their charisma and familiarity and are particularly valuable "ambassadors" of biodiversity conservation for public outreach. Here we present the data from our citizen science project, the first "Peninsular Malaysia Butterfly Count". Participants were asked to go outdoors on June 6, 2015, and (non-lethally) sample butterfly legs for species identification through DNA barcoding. Fifty-seven citizens responded to our adverts and registered to take part in the butterfly count with many registering on behalf of groups. Collectively the participants sampled 220 butterfly legs from 26 mostly urban and suburban sampling localities. These included our university campus, a highschool, several public parks and private residences. On the basis of 192 usable DNA barcodes, 43 species were sampled by the participants. The most sampled species was Appias olferna, followed by Junonia orithya and Zizina otis. Twenty-two species were only sampled once, five were only sampled twice, and four were only sampled three times. Three DNA barcodes could not be assigned species names. The sampled butterflies revealed that widely distributed, cosmopolitan species, often those recently arrived to the

  9. Gene expression profiling of Spodoptera frugiperda hemocytes and fat body using cDNA microarray reveals polydnavirus-associated variations in lepidopteran host genes transcript levels

    Directory of Open Access Journals (Sweden)

    Feyereisen R

    2006-06-01

    Full Text Available Abstract Background Genomic approaches provide unique opportunities to study interactions of insects with their pathogens. We developed a cDNA microarray to analyze the gene transcription profile of the lepidopteran pest Spodoptera frugiperda in response to injection of the polydnavirus HdIV associated with the ichneumonid wasp Hyposoter didymator. Polydnaviruses are associated with parasitic ichneumonoid wasps and are required for their development within the lepidopteran host, in which they act as potent immunosuppressive pathogens. In this study, we analyzed transcriptional variations in the two main effectors of the insect immune response, the hemocytes and the fat body, after injection of filter-purified HdIV. Results Results show that 24 hours post-injection, about 4% of the 1750 arrayed host genes display changes in their transcript levels with a large proportion (76% showing a decrease. As a comparison, in S. frugiperda fat body, after injection of the pathogenic JcDNV densovirus, 8 genes display significant changes in their transcript level. They differ from the 7 affected by HdIV and, as opposed to HdIV injection, are all up-regulated. Interestingly, several of the genes that are modulated by HdIV injection have been shown to be involved in lepidopteran innate immunity. Levels of transcripts related to calreticulin, prophenoloxidase-activating enzyme, immulectin-2 and a novel lepidopteran scavenger receptor are decreased in hemocytes of HdIV-injected caterpillars. This was confirmed by quantitative RT-PCR analysis but not observed after injection of heat-inactivated HdIV. Conversely, an increased level of transcripts was found for a galactose-binding lectin and, surprisingly, for the prophenoloxidase subunits. The results obtained suggest that HdIV injection affects transcript levels of genes encoding different components of the host immune response (non-self recognition, humoral and cellular responses. Conclusion This analysis of the

  10. Sea Star Wasting Disease in the Keystone Predator Pisaster ochraceus in Oregon: Insights into Differential Population Impacts, Recovery, Predation Rate, and Temperature Effects from Long-Term Research.

    Science.gov (United States)

    Menge, Bruce A; Cerny-Chipman, Elizabeth B; Johnson, Angela; Sullivan, Jenna; Gravem, Sarah; Chan, Francis

    2016-01-01

    Sea star wasting disease (SSWD) first appeared in Oregon in April 2014, and by June had spread to most of the coast. Although delayed compared to areas to the north and south, SSWD was initially most intense in north and central Oregon and spread southward. Up to 90% of individuals showed signs of disease from June-August 2014. In rocky intertidal habitats, populations of the dominant sea star Pisaster ochraceus were rapidly depleted, with magnitudes of decline in density among sites ranging from -2x to -9x (59 to 84%) and of biomass from -2.6x to -15.8x (60 to 90%) by September 2014. The frequency of symptomatic individuals declined over winter and persisted at a low rate through the spring and summer 2015 (~5-15%, at most sites) and into fall 2015. Disease expression included six symptoms: initially with twisting arms, then deflation and/or lesions, lost arms, losing grip on substrate, and final disintegration. SSWD was disproportionally higher in orange individuals, and higher in tidepools. Although historically P. ochraceus recruitment has been low, from fall 2014 to spring 2015 an unprecedented surge of sea star recruitment occurred at all sites, ranging from ~7x to 300x greater than in 2014. The loss of adult and juvenile individuals in 2014 led to a dramatic decline in predation rate on mussels compared to the previous two decades. A proximate cause of wasting was likely the "Sea Star associated Densovirus" (SSaDV), but the ultimate factors triggering the epidemic, if any, remain unclear. Although warm temperature has been proposed as a possible trigger, SSWD in Oregon populations increased with cool temperatures. Since P. ochraceus is a keystone predator that can strongly influence the biodiversity and community structure of the intertidal community, major community-level responses to the disease are expected. However, predicting the specific impacts and time course of change across west coast meta-communities is difficult, suggesting the need for detailed

  11. Investigation on arboviruses at Sino-Vietnam border areas in Wenshan of Yunnan province%云南省文山中越边境地区虫媒病毒调查

    Institute of Scientific and Technical Information of China (English)

    周涛; 张海林; 李铭华; 王静林; 付士红; 冯云; 梁国栋

    2009-01-01

    Objective To investigate arboviruses in Wenshan and Hekou county which are the Sino-Vietnam frontier regions of Wenshan, Yunnan province, China. Methods In September 2007,6091 culicines, 1334 anophelines, 848 aedes vexans and 53 armigeres obturbans were collected from 5 field sites. Mosquitoes were collected and stored in liquid nitrogen after classification. The mosquito pools were homogenized,and centrifuged,then the supernatant was inoculated onto C6/ 36 and BHK-21 cells,and the viral. isolates were identified by serological. and molecular biological. methods. Sequence alignment and phylogenetic analysis on the viral. isolates were carried out using Clustal. X 1. 85,GENEDOC and MEGA4 software. Results A total. of 4 pairs of virus isolated with C6/36 cells cytopathic effect were observed,and other mosquito species have not cytopathic effect Strain WS0704-2 was Banna virus which identified by antibody response and PCR. Strain WS0704-1, WS0708-1, WS0708-2 were culex pipens pallens densovirus (CppDNV) which identified by PCR. The phylogenetic analysis the 12th segment showed significant difference between the new banna virus and other strains isolated in China. Conclusion There are many mosquito vectors in frontier regions (China and Vietnam) of Wenshan in Yunnan province of China,and mosquito-borne arbovirus,such as BAV were isolated here.%目的 对云南省文山县、河口县等中越边境地区开展虫媒病毒调查,以期了解当地蚊虫携带虫媒病毒情况.方法 2007年9月在当地5个采集点共采集蚊虫8326只,包括库蚊6091只,中华按蚊1334只,刺扰伊蚊848只,阿蚊53只,并保存于液氮.经消毒、研磨、离心等处理后进行组织培养细胞接种,分离病毒和对病毒分离物进行血清学和分子生物学鉴定,以软件进行病毒的核苷酸序列比对和系统发生分析.结果 从库蚊分离到4株对C6/36细胞致病变的病毒分离物,其他蚊种没有分离到病毒分离

  12. 云南省东北等地区蚊虫及蚊媒病毒调查研究%Investigation of mosquitoes and mosquito-borne viruses in northeast Yunnan province

    Institute of Scientific and Technical Information of China (English)

    杨杜鹃; 梁国栋; 付士红; 张海林; 杨卫红; 冯云; 王静林; 章域震; 王丕玉; 陈维欣

    2011-01-01

    Objective To investigate the distribution patterns of mosquitoes and mosquito - borne viruses in Yunnan province to provide evidence for prevention and control of mosquito-borne diseases. Methods In 2009, mosquito samples were collected from 6 counties of Yunnan province. Mosquitoes were first classified, followed by cell culturing for viral isolation for molecular identification. Positive isolates were identified using SDS-PAGE and RT-PCR. The genomes of isolates were also, sequenced for phylogenetic analysis. Results A total of 18 562 mosquitos of 4 genera (Cidex, Anopheles, Armigeres, Aedes) and 24 species were collected. Cx. Tritaeniorhynchus and An. Sinensis were the major species, comprising 58.37% and 28.45% of the total population, respectively. Fifteen strains of viruses were isolated from the mosquito pools. RT-PCR and phylogenetic analysis revealed two viral strains from Cx. Tritaeniorhynchus, identified as Japanese encephalitis virus (JEV, Genotype I). One strain isolated from An. Sinensis was identified as Banna virus. Twelve vrial strains were identified as Cx. Pipiens pollens densovirus (CppDNV), of these, nine strains were isolated from Cx. Tritaeniorhynchus, and three strains from An. Sinensis. Conclusion Cx. Tritaeniorhynchus and An. Sinensis were the predominant species in the investigated areas. JEV, Banna virus and CppDNV were isolated. In northeast Yunnan, JEV was isolated for the first time.%目的 了解滇东北等地区蚊虫及蚊传虫媒病毒分布特点,为虫媒病毒病防治提供科学依据.方法 2009年在滇东北等地区的6个县采集蚊虫标本;蚊虫经分类鉴定后,用细胞培养法分离病毒,对病毒分离物进行分子生物学鉴定.结果 共采集到4属(库蚊、按蚊、阿蚊、伊蚊)24种18 562只蚊虫,其中三带喙库蚊、中华按蚊为主要蚊种,构成比分别为58.37%和28.45%;从蚊虫标本中分离到15株病毒,经分子生物学鉴定,其中2株(YN091l和YN0967)为基因Ⅰ型

  13. 山西省运城市2012年蚊媒病毒的分离鉴定%Isolation and identification of mosquito-borne arboviruses in Yuncheng city, Shanxi province, 2012

    Institute of Scientific and Technical Information of China (English)

    郑雅匀; 曹玉玺; 付士红; 程璟侠; 赵俊英; 代培芳; 孔祥盛; 梁国栋

    2015-01-01

    Objective To investigate the species and distribution of mosquitoes and mosquito-borne arboviruses in Yuncheng city of Shanxi province,China.Methods Mosquito samples were collected in 19 collection sites from Linyi county and Yongji city in Yuncheng city,in August,2012.After identification and classification,all the specimens were homogenized and centrifuged to acquire supernatant before being inoculated to both C6/36 and BHK21 cells for viral isolation.Positive isolates were identified with arbovirus species-specific primers under RT-PCR,for further sequencing and phylogenetic analysis.Results A total of 10 455 mosquitoes of 7 species in 4 genuese were collected.The predominant mosquito species in Linyi county was Culex pipens pallens (91.96%,3 911/4 253),but the one in Yongji city was Culex tritaeniorhynchus (72.85%,4 518/6 202).A total of 23 strains of viruses were isolated from the mosquito pools.15 strains from Culex tritaeniorhynchus and Culex pipens pallens were identified as genotype I Japanese encephalitis virus (JEV).Four strains from Culex pipens pallens were identified as Culex flavivirus (CxFV).Three strains from Culex pipens pallens were identified as Culex pipiens pallens densovirus (CppDNV).One strain from Armigeres subalbatus and Aedes albopictus was identified as Getah virus (GETV).Conclusion Four kinds of arboviruses were isolated from the mosquito pools,including GETV and CxFV,which were isolated and documented in Shanxi province for the first time.In the city of Yuncheng,Culex tritaeniorhynchus had been the predominant species and major vector for transmitting JEV.Genotype I JEV remained the major JEV circulating in the local natural environment.%目的 了解山西省运城市蚊虫及蚊媒病毒的种类和分布.方法 2012年8月在山西省运城市临猗县和永济市采集蚊虫标本,经鉴定分类和分批研磨后,利用细胞(C6/36和BHK21)培养方法分离病毒,对阳性分离物使用蚊媒病毒种属特异引

  14. 云南省德宏州2007年和2010年蚊虫及蚊媒病毒调查%Investigation on mosquitoes and mosquito-borne viruses in Dehong prefecture,Yunnan province,2007 and 2010

    Institute of Scientific and Technical Information of China (English)

    冯云; 李轼; 张保森; 尹正留; 董兴齐; 王环宇; 梁国栋; 张海林; 付士红; 杨卫红; 章域震; 王丕玉; 杨捷; 刘永华; 董朝良

    2014-01-01

    目的:调查云南省德宏州蚊虫和蚊媒病毒分布特点。方法2007年和2010年的7月在德宏州5个县市利用诱蚊灯采集蚊虫标本,采用C6/36和BHK-21细胞分离病毒,RT-PCR和序列分析方法鉴定病毒分离物。结果共采集到6属29种43634只蚊虫,其中三带喙库蚊构成比高达78.69%,中华按蚊构成比为14.77%,其他蚊种数量较少。从蚊虫中分离到6株病毒,经RT-PCR和进化分析表明,其中3株为基因Ⅰ型流行性乙型脑炎病毒(JEV),均分离自三带喙库蚊;1株盖塔病毒(GETV)分离自三带喙库蚊;2株淡色库蚊浓核病毒(CppDNV)分别分离自三带喙库蚊和迷糊按蚊。结论三带喙库蚊是德宏州优势蚊种和蚊媒病毒主要传播媒介,当地存在基因Ⅰ型JEV、GETV和CppDNV的流行,并与以往云南省及我国其他地区的分离株有较近亲缘关系。%Objective To investigate the distribution patterns of mosquitoes and mosquito-borne viruses in Dehong prefecture,Yunnan province,China. Methods Mosquito samples were collected using the mosquito traps from five counties of Dehong prefecture on July,2007 and 2010. Mosquitoes were cell cultured for viral isolation,and positive isolates were identified using RT-PCR and sequence analysis. Results A total of 43 634 mosquitoes comprised of 29 species representing six genera were collected. Culex tritaeriorhynchus and Anopheles sinensis comprised 78.69% and 14.77% of the total. Six strains of viruses were isolated from the mosquito pools. RT-PCR and phylogenetic analysis revealed three strains from Cx. tritaeriorhynchus,identified as genotypeⅠJapanese encephalitis virus(JEV). One strain was identified from Cx. tritaeriorhynchus,as Getah virus (GETV). Two strains isolated from Cx. tritaeriorhynchus and Anopheles vagus were identified as Culex pipiens pallens Densovirus(CppDNV). Conclusion Cx. tritaeriorhynchus had been the major species of mosquitoes and mainly transmitting

  15. Investigation of Arboviruses Carried by Mosquitoes at Ports in Five Provinces of South China%南方五省口岸蚊类携带虫媒病毒调查

    Institute of Scientific and Technical Information of China (English)

    郑夔; 黄吉城; 李小波; 洪烨; 师永霞; 幸芦琴; 相大鹏; 郭波旋; 胡龙飞

    2009-01-01

    effect of 42 samples can be observed in C6/36 cell. And the RT-PCR assay to detect the flavivirus and alphavirus showed a result that no corresponding genetic fragments were amplified. One of the cell samples with typical cytopathie effect was selected to identify the unknown virus by random PCR amplification. A kind of densovirus hidden in C6/36 was found. Conclusion The mosquitoes collected at ports in five Provinces of the South China may not carry dengue virus, Chikungunya virus, West Nile virus and yellow fever virus, only a very low number of mosquitoes carried Japanese encephalitis virus. The genotype Ⅰ Japanese encephalitis virus carried with mosquito was found for the first time in Fujian Province. And the eytopathie effect obversed in C6/36 cell culture may be caused by a kind of Aedes albopictus C6/36 cell densovirus hidden in C6/36 cells.%[目的]掌握南方口岸蚊媒携带病毒的本底资料,为蚊传疾病的预防控制工作提供依据.[方法]采用电动吸蚊器人工法和捕蚊磁场自动法采集南方5省口岸各类蚊虫.采集到的蚊类超低温送至实验室,研磨处理后用荧光PCR方法检测登革病毒、乙脑病毒、黄热病毒、西尼罗病毒、基孔肯雅病毒等重要蚊媒病毒,结果阳性的标本进一步进行PCR扩增和核苷酸序列测定分析;蚊标本研磨液同时用C6/36细胞进行虫媒病毒分离培养,出现细胞病变后分别用黄病毒科、甲病毒科各自的通用引物进行鉴定;对未能鉴定的未知病毒进一步用随机PCR方法进行扩增、克隆、序列测定、Blast搜索.[结果]从南方5省口岸采集到各类蚊虫12575只,鉴定后共分成254组.各组标本经荧光PCR方法检测,结果登革病毒、黄热病毒、西尼罗病毒、基孔肯雅病毒均为阴性;检测到2份福建省来源三带喙库蚊的标本乙脑病毒核酸阳性,经乙脑病毒E基因引物PCR扩增、测序分析证实为G Ⅰ型病毒.254份标本经C6/36细胞分离培养出现42