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Sample records for spot virus isolado

  1. Characterization of Tomato yellow spot virus, a novel tomato-infecting begomovirus in Brazil Caracterização do Tomato yellow spot virus, um novo begomovírus isolado de tomateiro no Brasil

    Directory of Open Access Journals (Sweden)

    Renata Faier Calegario

    2007-09-01

    Full Text Available The objective of this work was the biological and molecular characterization of a begomovirus detected in São Joaquim de Bicas, Minas Gerais, Brazil, named TGV-[Bi2], by determining its host range, complete nucleotide sequence and phylogenetic relationships with other begomoviruses. Biological characterization consisted of a host range study using either sap inoculation or particle bombardment as inoculation methods. The yellow spot virus can infect plants in Solanaceae and Amaranthaceae, including economically importat crops as sweet pepper, and weeds as Datura stramonium and Nicotiana silvestris. For the molecular characterization, the full-length genome (DNA-A and DNA-B was amplified, cloned and completely sequenced. Sequence comparisons and phylogenetic analyses indicated that TGV-[Bi2] constitutes a novel begomovirus species named Tomato yellow spot virus (ToYSV, closely related to Sida mottle virus (SiMoV.O objetivo deste trabalho foi a caracterização biológica e molecular de um begomovírus detectado em tomateiros em São Joaquim de Bicas, Minas Gerais, denominado TGV-[Bi2]. A caracterização biológica consistiu em teste de gama de hospedeiros, realizado por meio de inoculação via extrato foliar tamponado ou bombardeamento de partículas. O isolado TGV-[Bi2] infecta plantas das famílias Solanaceae e Amaranthaceae, inclusive espécies economicamente importantes como o pimentão, e algumas plantas daninhas como Datura stramonium e Nicotiana silvestris. A caracterização molecular consistiu na clonagem e seqüenciamento de seu genoma completo (DNA-A e DNA-B. A comparação de seqüências e análise filogenética indicaram que o TGV-[Bi2] constitui uma nova espécie de begomovírus, denominada Tomato yellow spot virus (ToYSV, filogeneticamente relacionado ao Sida mottle virus (SiMoV.

  2. Restricted spread of tomato spotted wilt virus

    NARCIS (Netherlands)

    Maris, P.C.; Joosten, N.N.; Goldbach, R.W.; Peters, D.

    2003-01-01

    Spread of Tomato spotted wilt virus (TSWV) and population development of its vector Frankliniella occidentalis were studied on the pepper accessions CPRO-1 and Pikante Reuzen, which are resistant and susceptible to thrips, respectively. Viruliferous thrips were released on plants of each accession

  3. Characterization of a tailless white spot syndrome virus from ...

    African Journals Online (AJOL)

    Jane

    2011-10-17

    , Yangzhou, China. Accepted 15 July, 2011. In this study, a virus similar to the causative agent of white spot syndrome virus (WSSV) but without tail-like extension was identified and characterized from diseased Penaeus ...

  4. Clustering and cellular distribution characteristics of virus particles of Tomato spotted wilt virus and Tomato zonate spot virus in different plant hosts

    National Research Council Canada - National Science Library

    Zhang, Zhongkai; Zheng, Kuanyu; Dong, Jiahong; Fang, Qi; Hong, Jian; Wang, Xifeng

    2016-01-01

    Tomato spotted wilt virus (TSWV) and Tomato zonate spot virus (TZSV) are the two dominant species of thrip-transmitted tospoviruses, cause significant losses in crop yield in Yunnan and its neighboring provinces in China...

  5. Stability of Citrus tristeza virus protective isolates in field conditions Estabilidade de isolados protetores contra Citrus tristeza virus em condições de campo

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    Alessandra Tenório Costa

    2010-07-01

    Full Text Available The objective of this work was to monitor the maintenance of Citrus tristeza virus (CTV protective isolates stability in selected clones of 'Pêra' sweet orange (Citrus sinensis, preimmunized or naturally infected by the virus, after successive clonal propagations. The work was carried out in field conditions in the north of Paraná State, Brazil. Coat protein gene (CPG analysis of 33 isolates collected from 16 clones of 'Pêra' sweet orange was performed using single strand conformational polymorphism (SSCP. Initially, the isolates were characterized by symptoms of stem pitting observed in clones. Then viral genome was extracted and used as template for the amplification of CPG by reverse transcription polimerase chain reaction (RTPCR. RTPCR products electrophoretic profiles were analyzed using the Jaccard coefficient and the UPGMA method. The majority of the clones had weak to moderate stem pitting symptoms and its CTV isolates showed alterations in the SSCP profiles. However, the stability of the protective complex has been maintained, except for isolates from two analised clones. Low genetic variability was observed within the isolates during the studied years.O objetivo deste trabalho foi monitorar a manutenção da estabilidade de isolados protetores contra Citrus tristeza virus (CTV em clones selecionados de laranja 'Pêra' (Citrus sinensis pré-imunizados ou infectados naturalmente pelo vírus, após sucessivas propagações clonais. O trabalho foi realizado em condições de campo, no norte do Estado do Paraná. A análise do gene da capa protéica (GPC de 33 isolados, coletados de 16 clones de laranjeira 'Pêra', foi realizada com o uso da técnica polimorfismo conformacional da fita simples (SSCP. Inicialmente, os isolados foram caracterizados por meio de sintomas de caneluras observados nos clones. Em seguida, o genoma viral foi extraído e utilizado como molde para a amplificação do GCP com uso da transcrição reversa da rea

  6. The role of NSm during tomato spotted wilt virus infection

    NARCIS (Netherlands)

    Storms, M.M.H.

    1998-01-01

    In the past ten years the genome organisation of tomato spotted wilt virus (TSWV) has been intensively studied in our laboratory. Complete genome sequence data revealed that this enveloped plant virus belongs to the Bunyaviridae, a virus family further restricted to

  7. Generation and characterization of mutants of tomato spotted wilt virus

    NARCIS (Netherlands)

    Oliveira Resende, de R.

    1993-01-01

    In nature, tospoviruses like tomato spotted wilt virus (TSWV) are exclusively transmitted by thrips species (Sakimura, 1962) producing numerous enveloped virions during infection, which accumulate in the cisternae of the endoplasmatic. reticulum. system (Kitajima, 1965; Milne, 1970; Ie,

  8. Genetic variation among isolates of White Spot Syndrome Virus

    NARCIS (Netherlands)

    Marks, H.; Goldbach, R.W.; Vlak, J.M.; Hulten, van M.C.W.

    2004-01-01

    White spot syndrome virus (WSSV), member of a. new virus family called Nimaviridae, is a major scourge in worldwide shrimp, cultivation. Geographical isolates of WSSV identified so far are very similar in morphology and proteome, and show little difference in restriction fragment length polymorphism

  9. Multiple proteins of White spot syndrome virus involved in ...

    Indian Academy of Sciences (India)

    The recognition and attachment of virus to its host cell surface is a critical step for viral infection. Recent research revealed that -integrin was involved in White spot syndrome virus (WSSV) infection. In this study, the interaction of -integrin with structure proteins of WSSV and motifs involved in WSSV infection was ...

  10. Multiple proteins of White spot syndrome virus involved in ...

    Indian Academy of Sciences (India)

    2014-03-20

    Mar 20, 2014 ... The recognition and attachment of virus to its host cell surface is a critical step for viral infection. Recent research revealed that β-integrin was involved in White spot syndrome virus (WSSV) infection. In this study, the interaction of β-integrin with structure proteins of WSSV and motifs involved in WSSV ...

  11. Genomics and transcriptomics of White spot syndrome virus

    NARCIS (Netherlands)

    Marks, H.

    2005-01-01

    White Spot Syndrome Virus (WSSV) is a large enveloped DNA virus that infects shrimp and other crustaceans. The virions are approximately 275 x 120 nm in size and have an ovoid to bacilliform shape and a tail-like appendage at one end. Sequencing revealed that the circular, double stranded (ds) DNA

  12. Western flower thrips can transmit Tomato spotted wilt virus from virus-infected tomato fruits

    Science.gov (United States)

    Acquisition and transmission of Tomato spotted wilt virus from symptomatic tomato fruits by western flower thrips was demonstrated for the first time. This suggests that infected tomato fruits may be a source of virus and also provide an additional means of virus movement between geographic areas....

  13. Caracterização de um isolado de Bidens mosaic virus proveniente de alface Characterization of an isolate of Bidens mosaic virus (BiMV from lettuce

    Directory of Open Access Journals (Sweden)

    Gerson Shinia Suzuki

    2009-09-01

    Full Text Available Em 2004, plantas de alface com sintomas de mosaico coletadas em São Manuel - SP foram analisadas por microscopia eletrônica, constatando-se presença de partículas típicas de potyvirus com 730 nm de comprimento. Após purificação biológica por monolesionais em Chenopodium quinoa, o extrato vegetal foi inoculado em uma série de plantas diferenciadoras, verificando-se que o isolado testado foi capaz de infectar C. quinoa e C. amaranticolor induzindo lesões locais seguidas de mosaico sistêmico. Ervilha (Pisum sativum mostrou-se assintomática, e em diferentes cultivares de alface como Trocadero, White Boston, Regina, Verônica, Lucy Brown, Rafaela, Tainá, Vera e Laurel foi observado o mosaico. A cultivar Gizele foi tolerante ao vírus. O sequenciamento da região codificadora da proteína capsidial revelou maior identidade de aminoácidos (97% deste isolado com o Bidens mosaic virus - BiMV (nº de acesso AY960151. Diferentemente dos isolados de BiMV já descritos, este proveniente de alface não foi capaz de infectar Bidens pilosa, Helianthus annuus, Nicotiana tabacum TNN e N. glutinosa. A ocorrência natural do BiMV em alface, causando sintomas semelhantes aos do LMV e a suscetibilidade de várias das cultivares hoje plantadas, servem como um alerta para a correta diagnose do vírus a campo.In 2004 lettuce plants showing mosaic symptoms collected in São Manuel, SP were analyzed by electron microscopy, and particles with 730 nm typically from potyvirus were observed. After biological purification by monolesionals on Chenopodium quinoa, this isolate was sap inoculated on a host range assay. The virus infected C. quinoa and C. amaranticolor, causing local lesions and systemic mosaic. The virus did not induce symptoms on pea (Pisum sativum, but induced mosaic on the leaves of some lettuce cultivars such as Trocadero, White Boston, Regina, Verônica, Lucy Brown, Rafaela, Tainá, Vera and Laurel. The lettuce cultivar Gizele was tolerant to

  14. Molecular variability analyses of Apple chlorotic leaf spot virus ...

    Indian Academy of Sciences (India)

    The complete sequences of the coat protein (CP) gene of 26 isolates of Apple chlorotic leaf spot virus (ACLSV) from India were determined. The isolates were obtained from various pome (apple, pear and quince) and stone (plum, peach, apricot, almond and wild Himalayan cherry) fruit trees. Other previously characterized ...

  15. On the vaccination of shrimp against white spot syndrome virus

    NARCIS (Netherlands)

    Witteveldt, J.

    2006-01-01

    More than a decade after its discovery inSouth-East Asia, White Spot Syndrome Virus (WSSV) is still the most important (viral) pathogen in the shrimp culture industry. Despite the shift from culturingPenaeusmonodon

  16. Transgenic tomato hybrids resistant to tomato spotted wilt virus infection.

    NARCIS (Netherlands)

    Haan, de P.; Ultzen, T.; Prins, M.; Gielen, J.; Goldbach, R.; Grinsven, van M.

    1996-01-01

    Tomato spotted wilt virus (TSWV) infections cause significant economic losses in the commercial culture of tomato (Lycopersicon esculentum). Culture practices have only been marginally effective in controlling TSWV. The ultimate way to minimize losses caused by TSWV is resistant varieties. These can

  17. Tomato chlorotic spot virus Identified in Marsdenia floribunda in Florida

    Science.gov (United States)

    Ornamental crops including hoya, annual vinca and portulaca have recently been identified with Tomato chlorotic spot virus (TCSV) infections in Florida. Observations of Marsdenia floribunda, commonly known as Madagascar jasmine, in September 2016 revealed TCSV-like symptoms. Testing of these sympt...

  18. White Spot Syndrome Virus infection in Penaeus monodon is ...

    Indian Academy of Sciences (India)

    Home; Journals; Journal of Biosciences; Volume 38; Issue 5. White Spot Syndrome Virus infection in ... Molecular Biology Research Laboratory, Department of Zoology, Centre for Advance Studies, University of Pune, Pune 411 007, India; These authors contributed equally to this work. National Institute of Virology, Microbial ...

  19. Can white spot syndrome virus be transmitted through the ...

    African Journals Online (AJOL)

    The transmission of white spot syndrome virus (WSSV) in the aquatic environment by the pathway of phytoplankton through rotifer to artemia and shrimp was investigated. The phytoplankton Alexandrium tamarense and Alexandrium minutum were co-cultured with adult Fenneropenaeus chinensis infected with WSSV and ...

  20. Clustering and cellular distribution characteristics of virus particles of Tomato spotted wilt virus and Tomato zonate spot virus in different plant hosts

    OpenAIRE

    ZHANG Zhongkai; Zheng, Kuanyu; Dong, Jiahong; Fang, Qi; Hong, Jian; Wang, Xifeng

    2016-01-01

    Background Tomato spotted wilt virus (TSWV) and Tomato zonate spot virus (TZSV) are the two dominant species of thrip-transmitted tospoviruses, cause significant losses in crop yield in Yunnan and its neighboring provinces in China. TSWV and TZSV belong to different serogroup of tospoviruses but induce similar symptoms in the same host plant species, which makes diagnostic difficult. We used different electron microscopy preparing methods to investigate clustering and cellular distribution of...

  1. White spot syndrome virus: an overview on an emergent concern.

    Science.gov (United States)

    Sánchez-Paz, Arturo

    2010-01-01

    Viruses are ubiquitous and extremely abundant in the marine environment. One of such marine viruses, the white spot syndrome virus (WSSV), has emerged globally as one of the most prevalent, widespread and lethal for shrimp populations. However, at present there is no treatment available to interfere with the unrestrained occurrence and spread of the disease. The recent progress in molecular biology techniques has made it possible to obtain information on the factors, mechanisms and strategies used by this virus to infect and replicate in susceptible host cells. Yet, further research is still required to fully understand the basic nature of WSSV, its exact life cycle and mode of infection. This information will expand our knowledge and may contribute to developing effective prophylactic or therapeutic measures. This review provides a state-of-the-art overview of the topic, and emphasizes the current progress and future direction for the development of WSSV control strategies. © INRA, EDP Sciences, 2010.

  2. Transgenic tomato hybrids resistant to tomato spotted wilt virus infection.

    OpenAIRE

    de Haan,; Ultzen, T.; Prins, M; Gielen, J.; Goldbach, R; Grinsven, van, Saskia

    1996-01-01

    Tomato spotted wilt virus (TSWV) infections cause significant economic losses in the commercial culture of tomato (Lycopersicon esculentum). Culture practices have only been marginally effective in controlling TSWV. The ultimate way to minimize losses caused by TSWV is resistant varieties. These can be obtained by introgression of natural sources of resistance from wild relatives or by expressing viral sequences in transgenic tomato plants. We report high levels of resistance to TSWV obtained...

  3. El virus de la mancha clorótica del tomate: Tomato chlorotic spot virus (TCSV)

    Science.gov (United States)

    Tomato chlorotic spot virus has emerged as a major pathogen of vegetables in Puerto Rico, the Caribbean and Florida. This virus is transmitted by thrips making management difficult. Growers must be aware of the distribution, host range, insect vectors, symptoms, modes of transmission to successfully...

  4. Suscetibilidade de Spodoptera Frugiperda a isolados geográficos de um vírus de poliedrose nuclear Spodoptera Frugiperda susceptibility to nuclear polyhedrosis virus geographical isolates

    Directory of Open Access Journals (Sweden)

    Sérgio Arce Gomez

    1999-09-01

    Full Text Available O presente trabalho objetivou verificar a suscetibilidade de larvas de segundo ínstar de Spodoptera frugiperda (Smith, 1797 a sete isolados geográficos de um vírus de poliedrose nuclear (VPN, conduzindo-se sete bioensaios no Laboratório de Patologia de Insetos da Embrapa-Centro Nacional de Pesquisa de Soja, Londrina. Para cada isolado preparou-se dieta artificial contendo 0, 2x10³, 4x10³, 8x10³, 16x10³, 32x10³ e 64x10³ corpos poliédricos de inclusão (CPI/mL. Cada dose foi oferecida às larvas em copos de plástico de 50 mL, sob condições controladas (temperatura: 26±2ºC; umidade relativa: 60±10%; fotófase:14 horas. A análise (Probits realizada sobre o somatório de larvas mortas (contadas, diariamente, do quinto ao décimo quarto dia após a inoculação mostrou, com base na ausência de sobreposição das amplitudes dos intervalos de confiança das concentrações letais médias (CL50, que: o isolado de Sertaneja, PR (5.631 CPI/mL, foi o mais virulento; o da Guatemala (11.520 CPI/mL equivaleu aos de Ponta Grossa, PR (14.184 CPI/mL, Argentina (15.891 CPI/mL e Alabama, EUA (17.558 CPI/mL, mas foi superior aos isolados de Louisiana, EUA (19.325 CPI/mL e Sete Lagoas, MG (25.310 CPI/mL. A variação do tempo letal médio, de 8,3 a 10 dias, não foi significativa em relação aos isolados.In order to verify the Spodoptera frugiperda (Smith,1797 second instar larvae susceptibility to seven nuclear polyhedrosis virus (NPV geographical isolates, seven bioassays were carried out at Embrapa-Centro Nacional de Pesquisa de Soja, Insect Pathology Laboratory, Londrina, Paraná State, Brazil. Artificial diet containing 0 (control, 2x10³, 4x10³, 8x10³, 16x10³, 32x10³, and 64x10³ polyhedral inclusion bodies (PIB/mL was prepared for each virus isolate; each dose was offered, in 50 mL plastic cups to the larvae under controlled conditions (temperature 26±2ºC; relative humidity: 60±10% and photophase: 14 hours. The statistical analysis

  5. Deep insight into white spot syndrome virus vaccines: A review

    Directory of Open Access Journals (Sweden)

    MA Badhul Haq

    2012-02-01

    Full Text Available White spot syndrome virus (WSSV, the causative virus of the disease, is found in most shrimp farming areas of the world, where it causes large economic losses to the shrimp farming industry. The potentially fatal virus has been found to be a threat not only to all shrimp species, but also to other marine and freshwater crustaceans, such as crab and crayfish. To date, no effective prophylactic treatment measures are available for viral infections in shrimp and other crustaceans. Due to current aquaculture practices and the broad host range of WSSV, intervention strategies including vaccination against this virus would be pivotal to save and protect shrimp farming. Several achievements have been attained in the search of novel vaccines for WSSV. DNA vaccination, recombinant vaccines, oral vaccination techniques and gene therapy are some of the thrust areas of focus for scientists and researchers. This review article highlights the recent trends in the development of WSSV vaccines either as DNA vaccines or recombinant vaccines and their functioning strategies as suggested by the researchers worldwide.

  6. Phylogenetic analysis of Tomato mosaic virus from Hemerocallis sp. and Impatiens hawkeri Análise filogenética de Tomato mosaic virus isolado de Hemerocallis sp. e Impatiens hawkeri

    Directory of Open Access Journals (Sweden)

    Lígia Maria Lembo Duarte

    2007-12-01

    Full Text Available The culture and commercialization of ornamental plants have considerably increased in the last years. To supply the commercial demand, several Hemerocallis and Impatiens varieties have been bred for appreciated qualities such as flowers with a diversity of shapes and colors. With the aim of characterizing the tobamovirus isolated from Hemerocallis sp. (tobamo-H and Impatiens hawkeri (tobamo-I from the USA and São Paulo, respectively, as well as to establish phylogenetic relationships between them and other Tobamovirus species, the viruses were submitted to RNA extraction, RT-PCR amplification, coat-protein gene sequencing and phylogenetic analyses. Comparison of tobamovirus homologous sequences yielded values superior to 98.5% of identity with Tomato mosaic virus (ToMV isolates at the nucleotide level. In relation to tobamo-H, 100% of identity with ToMV from tomatoes from Australia and Peru was found. Based on maximum likelihood (ML analysis it was suggested that tobamo-H and tobamo-I share a common ancestor with ToMV, Tobacco mosaic virus, Odontoglossum ringspot virus and Pepper mild mottle virus. The tree topology reconstructed under ML methodology shows a monophyletic group, supported by 100% of bootstrap, consisting of various ToMV isolates from different hosts, including some ornamentals, from different geographical locations. The results indicate that Hemerocallis sp. and I. hawkeri are infected by ToMV. This is the first report of the occurrence of this virus in ornamental species in Brazil.O cultivo e comercialização de plantas ornamentais têm aumentado consideravelmente nos últimos anos. Para suprir a demanda comercial, diversas variedades de Hemerocallis sp. e Impatiens hawkeri têm sido desenvolvidas pelas qualidades apreciáveis como flores com diversidade de formas e cores. Com o objetivo de caracterizar o tobamovirus isolado de Hemerocallis sp. (tobamo-H e Impatiens hawkeri (tobamo-I provenientes dos EUA e São Paulo

  7. Reação de acessos de Lycopersicon spp. a um isolado de Potato Virus Y (PVYº de tomateiro Reaction of accesses of Lycopersicon spp. to an isolate of Potato Virus Y (PVYº in tomato

    Directory of Open Access Journals (Sweden)

    Silvia Regina Luz Palazzo

    2008-01-01

    Full Text Available Amostras de tomateiro (Lycopersicon esculentum Mill ‘Alambra’ coletadas nas regiões produtoras de Elias Fausto, Monte-Mor e Mogi-Guaçú (SP, com sintomas de amarelecimento foliar generalizado semelhante aqueles causados por vírus foram submetidas à identificação do agente causal, por testes biológicos de transmissão mecânica, pela determinação do círculo de hospedeiras, empregando-se plantas indicadoras e diferenciadoras pertencentes às famílias Chenopodiaceae e Solanaceae pela identificação sorológica por PTA-ELISA do Potato virus Y (PVY, Pepper yellow mosaic virus (PepYMV, Cucumber mosaic virus (CMV, Tomato mosaic virus (ToMV e DAS-ELISA com antissoros policlonais para as espécies Tomato spot wilt vírus (TSWV, Tomato chlorotic spot virus (TCSV, Groundnut ringspot virus (GRSV, Chrysanthemum stem necrosis virus (CSNV e anticorpos monoclonais para as estirpes do PVY comum (PVYº, PVY necrótico (PVY N e PVY clorótico (PVY C. Das amostras coletadas 19 reagiram positivamente, com o PVY em PTA-ELISA e PVYºem DAS-ELISA. Plantas de Chenopodium amaranticolor reagiram com sintoma local e plantas de Nicotiana glutinosa, N. tabacum ‘WB’, N. sylvestris, N.debneyi, N. tabacum ‘Sansun’ reagiram com sintomas de mosaico sistêmico; tomateiros ‘Alambra’ manifestaram sintomas de mosaico-amarelo. Plantas de Datura stramonium, D. metel e C. annuum ‘Magda’ não foram infectadas. A ausência de sintomas em C.annuum ‘Magda’, identificou a presença do PVY patotipo 1 (PVYº1, nas 19 amostras. Como foi identificada a mesma espécie de vírus nas amostras, optou-se pela inoculação do isolado de tomate ‘Alambra’ de Elias Fausto nos dezenove acessos de Lycopersicon spp. do Banco Ativo de Germoplasma (BAG- IAC. O delineamento dos experimentos foi inteiramente casualizado. A reação dos acessos foi avaliada pela manifestação dos sintomas, pelos resultados positivos ou negativos após testes de retro-inoculação e PTA

  8. Field-Usable Lateral Flow Immunoassay for the Rapid Detection of White Spot Syndrome Virus (WSSV)

    OpenAIRE

    Kulabhusan, Prabir Kumar; Rajwade, Jyutika M.; Sugumar, Vimal; Taju, Gani; Sahul Hameed, A. S.; Paknikar, Kishore M.

    2017-01-01

    Background White spot disease (WSD), a major threat to sustainable aquaculture worldwide, is caused by White spot syndrome virus (WSSV). The diagnosis of WSD relies heavily on molecular detection of the virus by one-step PCR. These procedures are neither field-usable nor rapid enough considering the speed at which the virus spreads. Thus, development of a rapid, reliable and field-usable diagnostic method for the detection of WSSV infection is imperative to prevent huge economic losses. Metho...

  9. In silico identification of putative promoter motifs of White Spot syndrome virus

    NARCIS (Netherlands)

    Marks, H.; Ren, X.Y.; Sandbrink, H.; Hulten, van M.C.W.; Vlak, J.M.

    2006-01-01

    Background: White Spot Syndrome Virus, a member of the virus family Nimaviridae, is a large dsDNA virus infecting shrimp and other crustacean species. Although limited information is available on the mode of transcription, previous data suggest that WSSV gene expression occurs in a coordinated and

  10. First report of Tomato chlorotic spot virus on Annual Vinca (Catharanthus roseus) in the United States

    Science.gov (United States)

    Tomato chlorotic spot virus was identified in the ornamental crop Catharanthus roseus (commonly known as vinca) in south Florida, the first report of this virus naturally infecting this species. Genetic diversity of the virus was characterized. This report provides an overview of this emerging vir...

  11. Variabilidade biológica de isolados do Citrus leprosis virus (CiLV oriundos de cultivares de laranjeira Biological variability of Citrus leprosis virus (CiLV isolates from sweet orange cultivars

    Directory of Open Access Journals (Sweden)

    Jadier de Oliveira Cunha Junior

    2007-09-01

    Full Text Available A leprose, causada pelo Citrus leprosis virus (CiLV, é uma das principais doenças presentes em pomares cítricos fluminenses. O objetivo deste trabalho foi comparar o quadro sintomatológico desenvolvido por isolados de CiLV obtidos de cultivares comerciais de laranjeira (Lima, Pêra e Seleta, inoculados mecanicamente em Chenopodium amaranticolor, em três diluições. Após cinco a sete dias da inoculação foram observadas lesões necróticas, com pequeno halo clorótico quando observadas contra a luz. O maior número de lesões, nas três diluições, foi obtido do isolado de 'Seleta', seguido por 'Pêra' e 'Lima'. A melhor diluição utilizada para a observação das lesões foi de 1:10. Os resultados demonstram uma possível variabilidade biológica entre os isolados virais e/ou uma menor ou maior replicação viral, dependendo da cultivar, indicando um possível mecanismo de resistência da planta ao vírus.Citrus leprosis, caused by Citrus leprosis virus (CiLV is one of the major diseases in citrus orchards in the State of Rio de Janeiro. The objective of this study was to compare the development of symptoms on Chenopodium amaranticolor inoculated with CiLV isolates from 'Lima', 'Pera' and 'Seleta' sweet oranges using three different dilutions. Five to seven-days after inoculation, necrotic lesions exhibiting a small chlorotic halo when exposed to the light, were observed. The highest number of lesions developed using the three dilutions was obtained from 'Seleta', an intermediate value from 'Pêra' and the lowest number from 'Lima'. The best dilution for lesions development was 1:10. The results demonstrate a possible biological variability among the virus isolates and/or a lower or higher viral replication, dependent on the cultivar. This indicates a putative mechanism of Citrus resistance to the virus.

  12. Phylogenetic and recombination analysis of tomato spotted wilt virus.

    Directory of Open Access Journals (Sweden)

    Sen Lian

    Full Text Available Tomato spotted wilt virus (TSWV severely damages and reduces the yield of many economically important plants worldwide. In this study, we determined the whole-genome sequences of 10 TSWV isolates recently identified from various regions and hosts in Korea. Phylogenetic analysis of these 10 isolates as well as the three previously sequenced isolates indicated that the 13 Korean TSWV isolates could be divided into two groups reflecting either two different origins or divergences of Korean TSWV isolates. In addition, the complete nucleotide sequences for the 13 Korean TSWV isolates along with previously sequenced TSWV RNA segments from Korea and other countries were subjected to phylogenetic and recombination analysis. The phylogenetic analysis indicated that both the RNA L and RNA M segments of most Korean isolates might have originated in Western Europe and North America but that the RNA S segments for all Korean isolates might have originated in China and Japan. Recombination analysis identified a total of 12 recombination events among all isolates and segments and five recombination events among the 13 Korea isolates; among the five recombinants from Korea, three contained the whole RNA L segment, suggesting reassortment rather than recombination. Our analyses provide evidence that both recombination and reassortment have contributed to the molecular diversity of TSWV.

  13. Tomato spotted wilt virus infection improves host suitability for its vector Frankliniella occidentalis

    NARCIS (Netherlands)

    Maris, P.C.; Joosten, N.N.; Goldbach, R.W.; Peters, D.

    2004-01-01

    The effect of Tomato spotted wilt virus (TSWV) infection on plant attractiveness for the western flower thrips (Frankliniella occidentalis) was studied. Significantly more thrips were recovered on infected than were recovered on noninfected pepper (Capsicum annuum) plants in different preference

  14. Trichoderma L-Lysine-α-Oxidase Producer Strain Culture Fluid Inhibits Impatiens Necrotic Spot Virus.

    Science.gov (United States)

    Smirnova, I P; Shneider, Yu A; Karimova, E V

    2016-01-01

    A method for PCR diagnosis of impatiens necrotic spot virus is developed. Concentrated culture fluid with active L-lysine-α-oxidase (0.54 U/ml) from Trichoderma harzianum Rifai fungus producer strain F-180 inhibits vitally hazardous impatiens necrotic spot phytovirus.

  15. Western flower thrips can transmit Tomato spotted wilt virus from infected tomato fruits

    Science.gov (United States)

    Tomato spotted wilt virus (TSWV) has long been known to spread via plant propagation materials including transplants. Global dissemination of TSWV has also been linked to transport of thrips-infested and virus-infected horticultural and floricultural products through trade and commerce. However, th...

  16. Virion composition and genomics of white spot syndrome virus of shrimp

    NARCIS (Netherlands)

    Hulten, van M.C.W.

    2001-01-01


    Since its first discovery in Taiwan in 1992, White spot syndrome virus (WSSV) has caused major economic damage to shrimp culture. The virus has spread rapidly through Asia and reached the Western Hemisphere in 1995 (Texas), where it continued its devastating effect

  17. Identification and phylogeny of a protein kinase gene of white spot syndrome virus

    NARCIS (Netherlands)

    Hulten, van M.C.W.; Vlak, J.M.

    2001-01-01

    White spot syndrome virus (WSSV) is a virus infecting shrimp and other crustaceans, which is unclassified taxonomically. A 2193 bp long open reading frame, encoding a putative protein kinase (PK), was found on a 8.4 kb EcoRI fragment of WSSV proximal to the gene for the major envelope protein

  18. Identification of two major virion protein genes of white spot syndrome virus of shrimp

    NARCIS (Netherlands)

    Hulten, van M.C.W.; Westenberg, M.; Goodall, S.D.; Vlak, J.M.

    2000-01-01

    White Spot Syndrome Virus (WSSV) is an invertebrate virus, causing considerable mortality in shrimp. Two structural proteins of WSSV were identified. WSSV virions are enveloped nucleocapsids with a bacilliform morphology with an approximate size of 275 x 120 nm, and a tail-like extension at one end.

  19. Three functionally diverged major White Spot Syndrome Virus structural proteins evolved by gene duplication

    NARCIS (Netherlands)

    Hulten, van M.C.W.; Goldbach, R.W.; Vlak, J.M.

    2000-01-01

    White spot syndrome virus (WSSV) is an invertebrate virus causing considerable mortality in penaeid shrimp. The oval-to-bacilliform shaped virions, isolated from infected Penaeus monodon, contain four major proteins: VP28, VP26, VP24 and VP19 (28, 26, 24 and 19 kDa, respectively). VP26 and VP24 are

  20. The route of tomato spotted wilt virus inside the thrips body in relation to transmission efficiency

    NARCIS (Netherlands)

    Kritzman, A.; Gera, A.; Raccah, B.; Lent, van J.W.M.; Peters, D.

    2002-01-01

    The route of tomato spotted wilt virus (TSWV) in the body of its vectors, Frankliniella occidentalis and Thrips tabaci (Thysanoptera: Thripidae) was studied during their development. First instar larvae were allowed, immediately upon hatching, to acquire virus from mechanically infected Datura

  1. Occurrence of Tomato spotted wilt virus in Stevia rebaudiana and Solanum tuberosum in Northern Greece

    NARCIS (Netherlands)

    Chatzivassiliou, E.K.; Peters, D.; Lolas, P.

    2007-01-01

    Tomato spotted wilt virus (TSWV) (genus Tospovirus, family Bunyaviridae) was first reported in Greece during 1972 (3) and currently is widespread in the central and northern part of the country infecting several cultivated and wild plant species (1,2). In June 2006, virus-like symptoms similar to

  2. First report of the natural occurrence of tomato chlorotic spot virus in peanuts in Haiti

    Science.gov (United States)

    Tomato chlorotic spot virus (TCSV) was identified in peanut in Haiti. This is the first report of TCSV naturally infecting peanut. Genetic diversity of TCSV was characterized. This report provides an overview of this emerging virus for growers, extension workers, crop consultants and research and...

  3. Pathogenicity of different rabies virus isolates and protection test in vaccinated mice Patogenicidade de diferentes isolados do vírus da raiva e teste de proteção em camundongos vacinados

    Directory of Open Access Journals (Sweden)

    Elenice M.S. Cunha

    2010-10-01

    Full Text Available This study was aimed to evaluate and compare the pathogenicity of rabies virus isolated from bats and dogs, and to verify the efficacy of a commercial rabies vaccine against these isolates. For evaluation of pathogenicity, mice were inoculated by the intramuscular route (IM with 500MICLD50/0.03mL of the viruses. The cross-protection test was performed by vaccinating groups of mice by the subcutaneous route and challenged through the intracerebral (IC route. Isolates were fully pathogenic when inoculated by the IC route. When inoculated intramuscularly, the pathogenicity observed showed different death rates: 60.0% for the Desmodus rotundus isolate; 50.0% for dog and Nyctinomops laticaudatus isolates; 40.0% for Artibeus lituratus isolate; 9.5% Molossus molossus isolate; and 5.2% for the Eptesicus furinalis isolate. Mice receiving two doses of the vaccine and challenged by the IC route with the isolates were fully protected. Mice receiving only one dose of vaccine were partially protected against the dog isolate. The isolates from bats were pathogenic by the IC route in mice. However, when inoculated through the intramuscular route, the same isolates were found with different degrees of pathogenicity. The results of this work suggest that a commercial vaccine protects mice from infection with bat rabies virus isolates, in addition to a canine rabies virus isolate.O estudo avaliou e comparou as propriedades patogênicas de cinco isolados do vírus da raiva de morcegos e um isolado do vírus da raiva de cão e analisou a eficácia de vacina comercial contra estes isolados, em camundongos. Para o estudo de patogenicidade camundongos foram inoculados pela via IM com 0,1 mL contendo 500MICLD50/0,03mL das amostras de vírus. Quando inoculados pela via IC, os isolados do vírus da raiva provocaram a morte de 100% dos camundongos. No entanto, 500MICLD50/0,03mL das mesmas amostras, inoculadas pela via IM, ocasionaram mortalidade de: 60,0% quando a amostra

  4. Characterization of a tailless white spot syndrome virus from ...

    African Journals Online (AJOL)

    WSSV) but without tail-like extension was identified and characterized from diseased Penaeus vannamei and moribund Procambarus clarkia. Contrary to previous reports, white spots were not observed on the carapace of the diseased P.

  5. Cytopathological characteristics of tomato spotted wilt virus isolates

    Directory of Open Access Journals (Sweden)

    Anna Rudzińska-Langwald

    2014-01-01

    Full Text Available The electron microscopy study revealed that four examined virus isolates in the cells of the infected host plant produced different inclusions depending on the virus isolate and the time of passaging by mechanical transmission. Numerous virus particle inclusions as well as viroplasm and filamentous inclusions typical for TSWV were present in the plant cells infected with TSWV isolate (PPR. This isolate was kept in N. rustica by 4 mechanical transmissions. A similar virus isolate but maintained for 2 years by mechanical transmission in Nicotiana plants (TI produced virus particle inclusions as well as amorphous inclusions typical for defective isolates. In plant cells infected with the same isolate but maintained by mechanical transmission one year longer (T2 no virus particle inclusions were produced. In the amorphous inclusions produced by this isolate virus particles were seen, but they were not surrounded by additional membrane. The isolate G induced only amorphous inclusions dispersed within the cytoplasm of infected cells. No virus particles were seen in the amorphous inclusions. The mechanical transmission of TSWV isolates in N. rustica plants reduced the number of virus particles present in the cytoplasm. The defectivenes of the isolate cause also the appearance of a new type of inclusion - the amorphous inclusions.

  6. Comparative analysis of chrysanthemum transcriptome in response to three RNA viruses: Cucumber mosaic virus, Tomato spotted wilt virus and Potato virus X.

    Science.gov (United States)

    Choi, Hoseong; Jo, Yeonhwa; Lian, Sen; Jo, Kyoung-Min; Chu, Hyosub; Yoon, Ju-Yeon; Choi, Seung-Kook; Kim, Kook-Hyung; Cho, Won Kyong

    2015-06-01

    The chrysanthemum is one of popular flowers in the world and a host for several viruses. So far, molecular interaction studies between the chrysanthemum and viruses are limited. In this study, we carried out a transcriptome analysis of chrysanthemum in response to three different viruses including Cucumber mosaic virus (CMV), Tomato spotted wilt virus (TSWV) and Potato virus X (PVX). A chrysanthemum 135K microarray derived from expressed sequence tags was successfully applied for the expression profiles of the chrysanthemum at early stage of virus infection. Finally, we identified a total of 125, 70 and 124 differentially expressed genes (DEGs) for CMV, TSWV and PVX, respectively. Many DEGs were virus specific; however, 33 DEGs were commonly regulated by three viruses. Gene ontology (GO) enrichment analysis identified a total of 132 GO terms, and of them, six GO terms related stress response and MCM complex were commonly identified for three viruses. Several genes functioning in stress response such as chitin response and ethylene mediated signaling pathway were up-regulated indicating their involvement in establishment of host immune system. In particular, TSWV infection significantly down-regulated genes related to DNA metabolic process including DNA replication, chromatin organization, histone modification and cytokinesis, and they are mostly targeted to nucleosome and MCM complex. Taken together, our comparative transcriptome analysis revealed several genes related to hormone mediated viral stress response and DNA modification. The identified chrysanthemums genes could be good candidates for further functional study associated with resistant to various plant viruses.

  7. Expression and Characterization of a Soluble Form of Tomato Spotted Wilt Virus Glycoprotein GN

    OpenAIRE

    Whitfield, Anna E.; Ullman, Diane E.; German, Thomas L

    2004-01-01

    Tomato spotted wilt virus (TSWV), a member of the Tospovirus genus within the Bunyaviridae, is an economically important plant pathogen with a worldwide distribution. TSWV is transmitted to plants via thrips (Thysanoptera: Thripidae), which transmit the virus in a persistent propagative manner. The envelope glycoproteins, GN and GC, are critical for the infection of thrips, but they are not required for the initial infection of plants. Thus, it is assumed that the envelope glycoproteins play ...

  8. Tomato spotted wilt virus Gc and N proteins interact in vivo

    NARCIS (Netherlands)

    Snippe, M.; Borst, J.W.; Goldbach, R.W.; Kormelink, R.J.M.

    2007-01-01

    Tomato spotted wilt virus (TSWV) virions consist of a nucleocapsid core surrounded by a membrane containing glycoproteins Gn and Gc. To unravel the protein interactions involved in the membrane acquisition of RNPs, TSWV nucleocapsid protein (N), Gn and Gc were expressed and analyzed in BHK21 cells.

  9. Tomato spotted wilt virus in Sri Lanka: Emerging problems of Tospovirues

    NARCIS (Netherlands)

    Widana Gamage, S.M.K.; Hassani Mehraban, A.; Peters, D.

    2015-01-01

    Tomato spotted wilt virus (TSWV), the type species of the genus Tospovirus has long been reported infecting economically important horticultural crops such as tomato, groundnut, pepper, potato and soybean, all over the world. In Sri Lanka, characteristic tospovirus symptoms such as bud necrosis,

  10. Evaluation of thrips resistance in pepper to control Tomato spotted wilt virus infection

    NARCIS (Netherlands)

    Maris, P.C.

    2004-01-01

    The objective of this study was to analyse the effect of thrips ( F. occidentalis ) resistance in pepper ( Capsicum ) on the spread of Tomato spotted wilt virus (TSWV). Results demonstrate that the rate of primary TSWV-infection is effectively limited in a thrips-resistant (TR) pepper crop compared

  11. Treatment of white spot syndrome virus (WSSV) in penaeid shrimp aquaculture using plant extract

    Digital Repository Service at National Institute of Oceanography (India)

    Achuthankutty, C.T.; Desai, U.M.

    The white spot syndrome virus (WSSV) is causing a serious concern and threat to the shrimp aquaculture production in India since 1994. In this paper, the result of a successful attempt made in formulating a plant extract for treating the WSSV...

  12. Characterization of two biologically distinct variants of Tomato spotted wilt virus

    Science.gov (United States)

    Significant economic losses result on a wide range of crops due to infection with Tomato spotted wilt virus (TSWV). In this study, two TSWV isolates, one from basil and a second from tomato, were established in a common plant host. Viral proteins were monitored over time, plant host ranges were comp...

  13. Current status of Tomato chlorotic spot virus in Florida and the Caribbean

    Science.gov (United States)

    Damaging outbreaks of Tomato chlorotic spot virus (TCSV), an emerging thrips-vectored tospovirus, and several invasive species of thrips are significantly impacting vegetable and other crops in Florida and the Caribbean. Host and geographic ranges of TCSV are continuing to expand in this region. Dev...

  14. Transmission of white spot syndrome virus (WSSV) from Dendronereis spp. (Peters) (Nereididae) to penaeid shrimp

    NARCIS (Netherlands)

    Haryadi, D.; Verreth, J.A.J.; Verdegem, M.C.J.; Vlak, J.M.

    2015-01-01

    Dendronereis spp. (Peters) (Nereididae) is a common polychaete in shrimp ponds built on intertidal land and is natural food for shrimp in traditionally managed ponds in Indonesia. White spot syndrome virus (WSSV), an important viral pathogen of the shrimp, can replicate in this polychaete (Desrina

  15. Evolutionary trajectory of white spot syndrome virus (WSSV) genome shrinkage during spread in Asia

    NARCIS (Netherlands)

    Zwart, M.P.; Bui Thi Minh Dieu,; Hemerik, L.; Vlak, J.M.

    2010-01-01

    Background - White spot syndrome virus (WSSV) is the sole member of the novel Nimaviridae family, and the source of major economic problems in shrimp aquaculture. WSSV appears to have rapidly spread worldwide after the first reported outbreak in the early 1990s. Genomic deletions of various sizes

  16. Molecular Diagnosis of Iris Yellow Spot Virus (IYSV) on Onion in Iran

    NARCIS (Netherlands)

    Beikzadeh, N.; Jafarpour, B.; Rouhani, H.; Peters, D.; Hassani-Mehraban, A.

    2012-01-01

    Viral symptoms indicative of Iris yellow spot virus (IYSV) were observed on onion in several fields near Chenaran in Khorasan Razavi Province. Mechanical inoculation of herbaceous hosts with onion sap extracts from symptomatic plants showed similar symptoms to those described for IYSV. The

  17. Role of the envelope glycoproteins in the infection cycle of tomato spotted wilt virus

    NARCIS (Netherlands)

    Kikkert, M.

    1999-01-01

    Tomato spotted wilt virus (TSWV) forms the type member of the genus Tospovirus , which today harbors more than twelve different species. TSWV is able to infect an enormous variety of different plants, to which it often causes devastating effects,

  18. Primeiro relato do Tomato spotted wilt virus (TSWV em Campanula medium L. no Brasil First report of Tomato spotted wilt virus (TSWV infecting Campanula medium in Brazil

    Directory of Open Access Journals (Sweden)

    Ricardo Gioria

    2010-06-01

    Full Text Available Plantas de campânula (Campanula medium exibindo mosaico e necrose foliar e anéis em flores foram coletadas em uma estufa comercial de flores na região de Atibaia, SP. Suspeitando de possível etiologia viral, amostras de tecido lesionado foram analisadas por ensaios de transmissão mecânica, microscopia eletrônica e sorologia. Todos os resultados apontaram para a presença do Tomato spotted wilt virus (TSWV como o responsável pelos sintomas. Esse é o primeiro relato deste patógeno em campânula no Brasil.Plants of bellflower (Campanula medium exhibiting symptoms of mosaic, leaf necrosis and flower ring spot were found in a commercial crop in a greenhouse in Atibaia, São Paulo State. Electron microscopy, serology and biological assays indicated the presence of Tomato spotted wilt virus (TSWV. This is the first report of this pathogen in bellflower plants in Brazil.

  19. Clustering and cellular distribution characteristics of virus particles of Tomato spotted wilt virus and Tomato zonate spot virus in different plant hosts.

    Science.gov (United States)

    Zhang, Zhongkai; Zheng, Kuanyu; Dong, Jiahong; Fang, Qi; Hong, Jian; Wang, Xifeng

    2016-01-19

    Tomato spotted wilt virus (TSWV) and Tomato zonate spot virus (TZSV) are the two dominant species of thrip-transmitted tospoviruses, cause significant losses in crop yield in Yunnan and its neighboring provinces in China. TSWV and TZSV belong to different serogroup of tospoviruses but induce similar symptoms in the same host plant species, which makes diagnostic difficult. We used different electron microscopy preparing methods to investigate clustering and cellular distribution of TSWV and TZSV in the host plant species. Negative staining of samples infected with TSWV and TZSV revealed that particles usually clustered in the vesicles, including single particle (SP), double particles clustering (DPC), triple particles clustering (TPC). In the immunogold labeling negative staining against proteins of TZSV, the antibodies against Gn protein were stained more strongly than the N protein. Ultrathin section and high pressure freeze (HPF)-electron microscopy preparations revealed that TSWV particles were distributed in the cisternae of endoplasmic reticulum (ER), filamentous inclusions (FI) and Golgi bodies in the mesophyll cells. The TSWV particles clustered as multiple particles clustering (MPC) and distributed in globular viroplasm or cisternae of ER in the top leaf cell. TZSV particles were distributed more abundantly in the swollen membrane of ER in the mesophyll cell than those in the phloem parenchyma cells and were not observed in the top leaf cell. However, TZSV virions were mainly present as single particle in the cytoplasm, with few clustering as MPC. In this study, we identified TSWV and TZSV particles had the distinct cellular distribution patterns in the cytoplasm from different tissues and host plants. This is the first report of specific clustering characteristics of tospoviruses particles as well as the cellular distribution of TSWV particles in the FI and globular viroplasm where as TZSV particles inside the membrane of ER. These results indicated that

  20. Epidemiology of spotted wilt disease of peanut caused by Tomato spotted wilt virus in the southeastern U.S.

    Science.gov (United States)

    Culbreath, A K; Srinivasan, R

    2011-08-01

    Spotted wilt disease of peanut (Arachis hypogaea) (SWP), caused by Tomato spotted wilt virus (TSWV) (genus Tospovirus, family Bunyaviridae), was first observed in Alabama, Florida, and Georgia in the late 1980s and rapidly became a major limiting factor for peanut production in the region. Tobacco thrips (Frankliniella fusca) and western flower thrips (Frankliniella occidentalis) both occur on peanut throughout the southeastern U.S., but F. fusca is the predominant species that reproduces on peanut, and is considered to be the more important vector. Several non-crop sources of potential primary vectors and TSWV inoculum have been identified, but their relative importance has not been determined. The peanut growing season in Alabama, Florida, and Georgia is from April through November, and 'volunteer' peanut plants can be present for much of the remainder of the year. Therefore peanut itself has huge potential for perpetuating both vector and virus. Symptoms are often evident within a few days of seedling emergence, and disease progress is often rapid within the first 50-60 days after planting. Based on destructive sampling and assays for TSWV, there is often a high incidence of asymptomatic infections even in peanut genotypes that produce few and mild symptoms of infection in the field. Severity of SWP epidemics fluctuates significantly from year to year. The variability has not been fully explained, but lower incidences have been associated with years categorized as "La Niña" in the El Niño-Southern Oscillation. Planting date can have a large effect on disease incidence within a location. This may be linked to the thrips reproductive cycle and environmental effects on the plant and plant-thrips-virus interactions. Row pattern, plant population, and in-furrow applications of phorate insecticide can also affect epidemics of SWP. Considerable progress has been made in developing cultivars with natural field resistance to TSWV. Use of cultivars with moderate field

  1. Genetic diversity of papaya ring spot virus in Puerto Rico

    Science.gov (United States)

    The Food and Agriculture Organization estimates that 20-40% of crop yield is lost due to pests and diseases. Viruses are agents that cause diseases which contribute greatly to the global yield loss. Because of this, food production is negatively affected, especially in the tropics. Carica papaya, co...

  2. Tomato chocolate spot virus, a member of a new torradovirus species that causes a necrosis-associated disease of tomato in Guatemala

    OpenAIRE

    Batuman, O; Kuo, Y.-W.; Palmieri, M; Rojas, M. R.; Gilbertson, R. L.

    2010-01-01

    Tomatoes in Guatemala have been affected by a new disease, locally known as ?mancha de chocolate? (chocolate spot). The disease is characterized by distinct necrotic spots on leaves, stems and petioles that eventually expand and cause a dieback of apical tissues. Samples from symptomatic plants tested negative for infection by tomato spotted wilt virus, tobacco streak virus, tobacco etch virus and other known tomato-infecting viruses. A virus-like agent was sap-transmitted from diseased tissu...

  3. Primeiro relato do Tomato spotted wilt virus (TSWV) em Campanula medium L. no Brasil First report of Tomato spotted wilt virus (TSWV) infecting Campanula medium in Brazil

    OpenAIRE

    Ricardo Gioria; Kátia Regiane Brunelli; Romulo Fujito Kobori; Márcia Maria Rabelo Guimarães Kobori; Jorge Alberto Marques Rezende; Elliot Watanabe Kitajima

    2010-01-01

    Plantas de campânula (Campanula medium) exibindo mosaico e necrose foliar e anéis em flores foram coletadas em uma estufa comercial de flores na região de Atibaia, SP. Suspeitando de possível etiologia viral, amostras de tecido lesionado foram analisadas por ensaios de transmissão mecânica, microscopia eletrônica e sorologia. Todos os resultados apontaram para a presença do Tomato spotted wilt virus (TSWV) como o responsável pelos sintomas. Esse é o primeiro relato deste patógeno em campânula...

  4. Resistance breaking tomato spotted wilt virus isolates on resistant pepper varieties in Italy.

    Science.gov (United States)

    Crescenzi, A; Viggiano, A; Fanigliulo, A

    2013-01-01

    In spring 2012, resistance breaking (RB) isolates of tomato spotted wilt virus (TSWV) that overcome the resistance conferred by the Tsw gene in different pepper hybrids have been recovered in different locations in southern Italy (Campania and Apulia regions) in protected cultivation, about one month after transplant. The percentage of symptomatic plants was 5-10% and, only in particular cases of advanced stage of cultivation, it reached 30-50% at the end of cycle. All TSWV isolates induced similar systemic symptoms in all resistant infected pepper hybrids: yellowing or browning of apical leaves, which later become necrotic, long necrotic streakson stems, extending to the terminal shoots, complete necrosis of younger fruits and large necrotic streaks and spots on fruits formed after infection. On ripe fruits, yellow spots with concentric rings or necrotic streaks could be observed. Leaf extracts of these samples were tested in ELISA for the detection of TSWV, Cucumber mosaic virus (CMV), Tobacco mosaic virus (TMV), Tomato mosaic virus (ToMV), Impatiens necrotic spot virus (INSV), Potato virus Y (PVY), Alfalfa mosaic virus (AMV), Pepper mild mottle virus (PMMoV) and Pepper Mottle Virus (PepMoV). Only TSWV was detected in all the field samples tested. The correspondent virus isolates were inoculated mechanically and by Frankliniella occidentalis on to a set of different pepper and tomato hybrids, as well as on some herbaceous test plants, in order to investigate for their ability to overcome the resistance genes Tsw and Sw5, respectively. Tomato hybrids carrying the Sw5 gene were uninfected by all RB isolates, whereas all resistant pepper hybrids became systemically infected. RB isolates did not differ noticeably in transmission efficiency when they were tested with the thrips F. occidentalis. Obtained results demonstrate that evolved strains of TSWV have emerged, that they are able to overcome the Tsw resistance gene in pepper plants experimentally inoculated both

  5. Horizontal transmission dynamics of White spot syndrome virus by cohabitation trials in juvenile Penaeus monodon and P. vannamei

    NARCIS (Netherlands)

    Ngo Xuan, T.; Verreth, J.A.J.; Vlak, J.M.; Jong, de M.C.M.

    2014-01-01

    White spot syndrome virus (WSSV), a rod-shaped double-stranded DNA virus, is an infectious agent causing fatal disease in shrimp farming around the globe. Within shrimp populations WSSV is transmitted very fast, however, the modes and dynamics of transmission of this virus are not well understood.

  6. Cytopathological changes in Schefflera actinophylla Harms. naturally infected with impatiens necrotic spot virus (INSV

    Directory of Open Access Journals (Sweden)

    Anna Rudzińska-Langwald

    2014-01-01

    Full Text Available Plants of Schefflera actinophylla Harms. with stunted growth, chlorotic and necrotic spots and patterns, leaf epinasty and distortion are infected with impatiens necrotic spot virus classified as a member of genus Tospovirus. Studies with electron microscope revealed that in cells of S. actinophylla leaves there were no virus particle inclusions typical for tospovirus infection, but only single particles were present. The isolate of INSV was defective and the amount of INSV particles was strongly reduced in Schefflera plants. Some inclusions (I type were composed of short branched cisterns of endoplasmic reticulum and a dark substance forming characteristic bands. The II type represented inclusions that were usually ballshaped and composed of a dark substance resembling that of the I type, but not accompanied by endoplasmic reticulum cisterns. The III type of inclusions was composed of dark, osmophilic masses. The inclusions present in the cells differ from those typical for tospovirus infection.

  7. Effect of tiazofurin on tomato plants infected with tomato spotted wilt virus.

    Science.gov (United States)

    Caner, J; Amélia, M; Alexandre, V; Vicente, M

    1984-12-01

    Tiazofurin (2-beta-D-ribofuranosylthiazole-4-carboxamide) was examined for its activity against tomato-spotted wilt virus (TSWV) in tomato plants. Solutions containing 50, 100, 200, 400 and 800 mg/l of the drug were sprayed onto the leaves. The results showed that 100 and 200 mg/l were the most efficient concentrations to suppress TSWV infection, thereby delaying the appearance of systemic symptoms. The drug was more effective in controlling TSWV infection when applied after than before virus inoculation. The results suggest that tiazofurin can be used as an efficient antiviral drug in the treatment of TSWV-infected tomato plants.

  8. White spot syndrome virus infection: Threat to crustacean biodiversity in Vembanad Lake, India

    OpenAIRE

    Joseph, Toms C.; James, Roswin; Rajan, L. Anbu; Surendran, P.K.; Lalitha, K. V.

    2015-01-01

    The Vembanad Lake located on the south-west coast of India, an ecological hotspot is the nursing ground of many economically important crustaceans. The prevalence of white spot syndrome virus (WSSV) among crustaceans from farmed, estuarine and marine environments surrounding the Vembanad Lake, India was detected using PCR. A total of 308 samples from aquaculture ponds consisting of six species of crustaceans collected from five different farms were tested for the presence of WSSV. Of these, 6...

  9. Effectiveness of combining resistance to Thielaviopsis basicola and Tomato spotted wilt virus in haploid tobacco genotypes

    OpenAIRE

    Trojak-Goluch, Anna; Laskowska, Dorota; Agacka, Monika; Czarnecka, Diana; Kawka, Magdalena; Czubacka, Anna

    2011-01-01

    Black root rot (BRR) caused by Thielaviopsis basicola as well as Tomato spotted wilt virus (TSWV) are the most serious problems in tobacco growing regions. We crossed the breeding line WGL 3 carrying BRR resistance derived from N.glauca with the line PW-834 the resistance of which to TSWV was transferred from cultivar Polalta. Anthers obtained from F1 hybrid plants were cultured to induce haploids combining resistance to Th. basicola and TSWV. Flow cytometry analysis revealed 242 haploids and...

  10. Pattern of the Occurrence of Tomato spotted wilt virus in Jeonnam Province

    OpenAIRE

    Sug-Ju Ko; Beom-Ryong Kang; Duck-Soo Choi; Do-Ik Kim; Gwan-Seok Lee; Chang-Seok Kim; Hong-Soo Choi

    2013-01-01

    Tomato spotted wilt virus (TSWV) was occurred at 8 areas including Naju, Suncheon, Younggwang, Youngam, and Shinan in Jeonnam province and the crops of Younggwang were severely damaged by TSWV. The hot pepper (Capsicum annuum), bell pepper (Capsicum annuum v ar. angulosum) and tomato (Solanum lycopersicon) in greenhouse and hot pepper in open field were infected by TSWV. Especially, hot pepper was severely damaged by TSWV infection. The survey data indicated that 1.1−30% in the nu...

  11. Characterization of a New Tomato Spotted Wilt Virus Isolates Found in Hippeastrum hybridum (Hort.) Plants in Poland

    OpenAIRE

    Berniak Hanna

    2016-01-01

    Two Tomato spotted wilt virus (TSWV) isolates H1 and H2 found in Hippeastrum hybridum plants were characterized based on biological, serological, and molecular properties. Virus isolates showed differences in symptom expression – H1 isolate displayed severe necrotic spots and patterns, whereas mild mosaic symptoms were observed on H2-infected H. hybridum plants. Both TSWV isolates showed comparable reactivity with TSWV-specific antibodies and they induced similar symptoms on herbaceous indica...

  12. Emergence of Groundnut ringspot virus and Tomato chlorotic spot virus in Vegetables in Florida and the Southeastern United States.

    Science.gov (United States)

    Webster, Craig G; Frantz, Galen; Reitz, Stuart R; Funderburk, Joseph E; Mellinger, H Charles; McAvoy, Eugene; Turechek, William W; Marshall, Spencer H; Tantiwanich, Yaowapa; McGrath, Margaret T; Daughtrey, Margery L; Adkins, Scott

    2015-03-01

    Groundnut ringspot virus (GRSV) and Tomato chlorotic spot virus (TCSV) are two emerging tospoviruses in Florida. In a survey of the southeastern United States, GRSV and TCSV were frequently detected in solanaceous crops and weeds with tospovirus-like symptoms in south Florida, and occurred sympatrically with Tomato spotted wilt virus (TSWV) in tomato and pepper in south Florida. TSWV was the only tospovirus detected in other survey locations, with the exceptions of GRSV from tomato (Solanum lycopersicum) in South Carolina and New York, both of which are first reports. Impatiens (Impatiens walleriana) and lettuce (Lactuca sativa) were the only non-solanaceous GRSV and/or TCSV hosts identified in experimental host range studies. Little genetic diversity was observed in GRSV and TCSV sequences, likely due to the recent introductions of both viruses. All GRSV isolates characterized were reassortants with the TCSV M RNA. In laboratory transmission studies, Frankliniella schultzei was a more efficient vector of GRSV than F. occidentalis. TCSV was acquired more efficiently than GRSV by F. occidentalis but upon acquisition, transmission frequencies were similar. Further spread of GRSV and TCSV in the United States is possible and detection of mixed infections highlights the opportunity for additional reassortment of tospovirus genomic RNAs.

  13. Selection of tomato plants resistant to a local Polish isolate of tomato spotted wilt virus (TSWV).

    Science.gov (United States)

    Czech, Andrzej S; Szklarczyk, Marek; Gajewski, Zbigniew; Zukowska, Ewa; Michalik, Barbara; Kobyłko, Tadeusz; Strzałka, Kazimierz

    2003-01-01

    We found that the Sw-5 gene confers resistance to one of the Polish isolates of tomato spotted wilt virus (TSWV). A series of tomato breeding accessions was analysed along with standards of resistance and susceptibility to TSWV. The presence of the Sw-5 gene was determined using the available PCR marker. Subsequently plants from these accessions were grown in the presence of the TSWV isolate from Poland. Some of them developed severe symptoms of the TSWV disease. Expression of the virus proteins was also assayed in tissues of the investigated plants. We found general agreement between either lack or presence of the disease symptoms, virus proteins and resistance gene. Some observed discrepancies of these data are also discussed. Our results indicate that marker-assisted selection can be used for breeding of the TSWV-resistant tomato in Poland.

  14. ViroSpot microneutralization assay for antigenic characterization of human influenza viruses.

    Science.gov (United States)

    van Baalen, Carel A; Jeeninga, Rienk E; Penders, Germaine H W M; van Gent, Brenda; van Beek, Ruud; Koopmans, Marion P G; Rimmelzwaan, Guus F

    2017-01-03

    The hemagglutination inhibition (HI) assay has been used for the antigenic characterization of influenza viruses for decades. However, the majority of recent seasonal influenza A viruses of the H3N2 subtype has lost the capacity to agglutinate erythrocytes of various species. The hemagglutination (HA) activity of other A(H3N2) strains is generally sensitive to the action of the neuraminidase inhibitor oseltamivir, which indicates that the neuraminidase and not the hemagglutinin is responsible for the HA activity. These findings complicate the antigenic characterization and selection of A(H3N2) vaccine strains, calling for alternative antigenic characterization assays. Here we describe the development and use of the ViroSpot microneutralization (MN) assay as a reliable and robust alternative for the HI assay. Serum neutralization of influenza A(H3N2) reference virus strains and epidemic isolates was determined by automated readout of immunostained cell monolayers, in a format designed to minimize the influence of infectious virus doses on serum neutralization titers. Neutralization of infection was largely independent from rates of viral replication and cell-to-cell transmission, facilitating the comparison of different virus isolates. Other advantages of the ViroSpot MN assay include its relative insensitivity to variation in test dose of infectious virus, automated capture and analyses of residual infection patterns, and compatibility with standardized large scale analyses. Using this assay, a number of epidemic influenza A(H3N2) strains that failed to agglutinate erythrocytes, were readily characterized antigenically. Copyright © 2016 The Authors. Published by Elsevier Ltd.. All rights reserved.

  15. Field-Usable Lateral Flow Immunoassay for the Rapid Detection of White Spot Syndrome Virus (WSSV).

    Science.gov (United States)

    Kulabhusan, Prabir Kumar; Rajwade, Jyutika M; Sugumar, Vimal; Taju, Gani; Sahul Hameed, A S; Paknikar, Kishore M

    2017-01-01

    White spot disease (WSD), a major threat to sustainable aquaculture worldwide, is caused by White spot syndrome virus (WSSV). The diagnosis of WSD relies heavily on molecular detection of the virus by one-step PCR. These procedures are neither field-usable nor rapid enough considering the speed at which the virus spreads. Thus, development of a rapid, reliable and field-usable diagnostic method for the detection of WSSV infection is imperative to prevent huge economic losses. Here, we report on the development of a lateral flow immunoassay (LFIA) employing gold nanoparticles conjugated to a polyclonal antibody against VP28 (envelope protein of WSSV). The LFIA detected WSSV in ~20 min and showed no cross-reactivity with other shrimp viruses, viz. Monodon Baculovirus (MBV), Hepatopancreatic parvovirus (HPV) and Infectious Hypodermal and Hematopoietic Necrosis virus (IHHNV). The limit of detection (LOD) of the assay, as determined by real-time PCR, was 103 copies of WSSV. In a time course infectivity experiment, ~104 WSSV particles were injected in Litopenaeus vannamei. The LFIA could rapidly (~ 20 min) detect the virus in different tissues after 3 h (hemolymph), 6 h (gill tissue) and 12 h (head soft tissue, eye stalk, and pleopod) of infection. Based on these findings, a validation study was performed using 75 field samples collected from different geographical locations in India. The LFIA results obtained were compared with the conventional "gold standard test", viz. one-step PCR. The analysis of results in 2x2 matrix indicated very high sensitivity (100%) and specificity (96.77%) of LFIA. Similarly, Cohen's kappa coefficient of 0.983 suggested "very good agreement" between the developed LFIA and the conventional one-step PCR. The LFIA developed for the rapid detection of WSSV has an excellent potential for use in the field and could prove to be a boon to the aquaculture industry.

  16. Transmission of Iris yellow spot virus by Frankliniella fusca and Thrips tabaci (Thysanoptera: Thripidae).

    Science.gov (United States)

    Srinivasan, Rajagopalbabu; Sundaraj, Sivamani; Pappu, Hanu R; Diffie, Stan; Riley, David G; Gitaitis, Ron D

    2012-02-01

    Thrips-transmitted Iris yellow spot virus (IYSV) (Family Bunyaviridae, Genus Tospovirus) affects onion production in the United States and worldwide. The presence of IYSV in Georgia was confirmed in 2003. Two important thrips species that transmit tospoviruses, the onion thrips (Thrips tabaci (Lindeman)) and the tobacco thrips (Frankliniella fusca (Hinds)) are known to infest onion in Georgia. However, T. tabaci is the only confirmed vector of IYSV. Experiments were conducted to test the vector status of F. fusca in comparison with T. tabaci. F. fusca and T. tabaci larvae and adults reared on IYSV-infected hosts were tested with antiserum specific to the nonstructural protein of IYSV through an antigen coated plate ELISA. The detection rates for F. fusca larvae and adults were 4.5 and 5.1%, respectively, and for T. tabaci larvae and adults they were 20.0 and 24.0%, respectively, indicating that both F. fusca and T. tabaci can transmit IYSV. Further, transmission efficiencies of F. fusca and T. tabaci were evaluated by using an indicator host, lisianthus (Eustoma russellianum (Salisbury)). Both F. fusca and T. tabaci transmitted IYSV at 18.3 and 76.6%, respectively. Results confirmed that F. fusca also can transmit IYSV but at a lower efficiency than T. tabaci. To attest if low vector competency of our laboratory-reared F. fusca population affected its IYSV transmission capability, a Tomato spotted wilt virus (Family Bunyaviridae, Genus Tospovirus) transmission experiment was conducted. F. fusca transmitted Tomato spotted wilt virus at a competent rate (90%) suggesting that the transmission efficiency of a competent thrips vector can widely vary between two closely related viruses.

  17. The role of weeds in the spread of Tomato spotted wilt virus by thrips tabaci (Thysanoptera: Thripidae) in tobacco crops

    NARCIS (Netherlands)

    Chatzivassiliou, E.K.; Peters, D.; Katis, N.I.

    2007-01-01

    Oviposition of Thrips tabaci, larval development and their potential to acquire Tomato spotted wilt virus (TSWV) from infected Amaranthus retroflexus, Datura stramonium, Lactuca serriola, Solanum nigrum and Sonchus oleraceus plants and the ability of the adults to transmit this virus to these weeds

  18. Protein profiling in the gut of Penaeus monodon gavaged with oral WSSV-vaccines and live white spot syndrom virus

    NARCIS (Netherlands)

    Kulkarni, A.D.; Kiron, V.; Rombout, J.H.W.M.; Brinchmann, M.; Fernandes, J.M.O.; Sudheer, N.S.; Singh, B.I.S.

    2014-01-01

    White spot syndrome virus (WSSV) is a pathogen that causes considerable mortality of the farmed shrimp, Penaeus monodon. Candidate ‘vaccines’, WSSV envelope protein VP28 and formalin-inactivated WSSV, can provide short-lived protection against the virus. In this study, P. monodon was orally

  19. siRNA injection induces sequence-independent protection in Penaeus monodon against white spot syndrome virus

    NARCIS (Netherlands)

    Westenberg, M.; Heinhuis, B.; Zuidema, D.; Vlak, J.M.

    2005-01-01

    White spot syndrome virus (WSSV) is a major disease in crustaceans, particularly shrimp, due to the current intensity of aquaculture practices. Novel strategies including vaccination to control this virus would be highly desirable. However, invertebrates lack a true adaptive immune response system

  20. Editorial: Identification and incidence of iris yellow spot virus, a new pathogen in onion and leek in Greece

    NARCIS (Netherlands)

    Chatzivassiliou, E.K.; Giavachtsia, V.; Hassani-Mehraban, A.; Hoedjes, K.; Peters, D.

    2009-01-01

    Iris yellow spot virus (IYSV; genus Tospovirus, family Bunyaviridae) is an emerging and serious pathogen affecting several Allium spp. worldwide (2). The virus causes straw-colored, chlorotic or necrotic lesions that coalesce, occasionally resulting in an extensive necrosis on onion (A. cepa L.)

  1. Predictive Models for Tomato Spotted Wilt Virus Spread Dynamics, Considering Frankliniella occidentalis Specific Life Processes as Influenced by the Virus.

    Directory of Open Access Journals (Sweden)

    Pamella Akoth Ogada

    Full Text Available Several models have been studied on predictive epidemics of arthropod vectored plant viruses in an attempt to bring understanding to the complex but specific relationship between the three cornered pathosystem (virus, vector and host plant, as well as their interactions with the environment. A large body of studies mainly focuses on weather based models as management tool for monitoring pests and diseases, with very few incorporating the contribution of vector's life processes in the disease dynamics, which is an essential aspect when mitigating virus incidences in a crop stand. In this study, we hypothesized that the multiplication and spread of tomato spotted wilt virus (TSWV in a crop stand is strongly related to its influences on Frankliniella occidentalis preferential behavior and life expectancy. Model dynamics of important aspects in disease development within TSWV-F. occidentalis-host plant interactions were developed, focusing on F. occidentalis' life processes as influenced by TSWV. The results show that the influence of TSWV on F. occidentalis preferential behaviour leads to an estimated increase in relative acquisition rate of the virus, and up to 33% increase in transmission rate to healthy plants. Also, increased life expectancy; which relates to improved fitness, is dependent on the virus induced preferential behaviour, consequently promoting multiplication and spread of the virus in a crop stand. The development of vector-based models could further help in elucidating the role of tri-trophic interactions in agricultural disease systems. Use of the model to examine the components of the disease process could also boost our understanding on how specific epidemiological characteristics interact to cause diseases in crops. With this level of understanding we can efficiently develop more precise control strategies for the virus and the vector.

  2. Molecular Mechanisms of White Spot Syndrome Virus Infection and Perspectives on Treatments.

    Science.gov (United States)

    Verbruggen, Bas; Bickley, Lisa K; van Aerle, Ronny; Bateman, Kelly S; Stentiford, Grant D; Santos, Eduarda M; Tyler, Charles R

    2016-01-18

    Since its emergence in the 1990s, White Spot Disease (WSD) has had major economic and societal impact in the crustacean aquaculture sector. Over the years shrimp farming alone has experienced billion dollar losses through WSD. The disease is caused by the White Spot Syndrome Virus (WSSV), a large dsDNA virus and the only member of the Nimaviridae family. Susceptibility to WSSV in a wide range of crustacean hosts makes it a major risk factor in the translocation of live animals and in commodity products. Currently there are no effective treatments for this disease. Understanding the molecular basis of disease processes has contributed significantly to the treatment of many human and animal pathogens, and with a similar aim considerable efforts have been directed towards understanding host-pathogen molecular interactions for WSD. Work on the molecular mechanisms of pathogenesis in aquatic crustaceans has been restricted by a lack of sequenced and annotated genomes for host species. Nevertheless, some of the key host-pathogen interactions have been established: between viral envelope proteins and host cell receptors at initiation of infection, involvement of various immune system pathways in response to WSSV, and the roles of various host and virus miRNAs in mitigation or progression of disease. Despite these advances, many fundamental knowledge gaps remain; for example, the roles of the majority of WSSV proteins are still unknown. In this review we assess current knowledge of how WSSV infects and replicates in its host, and critique strategies for WSD treatment.

  3. Molecular Mechanisms of White Spot Syndrome Virus Infection and Perspectives on Treatments

    Directory of Open Access Journals (Sweden)

    Bas Verbruggen

    2016-01-01

    Full Text Available Since its emergence in the 1990s, White Spot Disease (WSD has had major economic and societal impact in the crustacean aquaculture sector. Over the years shrimp farming alone has experienced billion dollar losses through WSD. The disease is caused by the White Spot Syndrome Virus (WSSV, a large dsDNA virus and the only member of the Nimaviridae family. Susceptibility to WSSV in a wide range of crustacean hosts makes it a major risk factor in the translocation of live animals and in commodity products. Currently there are no effective treatments for this disease. Understanding the molecular basis of disease processes has contributed significantly to the treatment of many human and animal pathogens, and with a similar aim considerable efforts have been directed towards understanding host–pathogen molecular interactions for WSD. Work on the molecular mechanisms of pathogenesis in aquatic crustaceans has been restricted by a lack of sequenced and annotated genomes for host species. Nevertheless, some of the key host–pathogen interactions have been established: between viral envelope proteins and host cell receptors at initiation of infection, involvement of various immune system pathways in response to WSSV, and the roles of various host and virus miRNAs in mitigation or progression of disease. Despite these advances, many fundamental knowledge gaps remain; for example, the roles of the majority of WSSV proteins are still unknown. In this review we assess current knowledge of how WSSV infects and replicates in its host, and critique strategies for WSD treatment.

  4. Characterization of Melon necrotic spot virus Occurring on Watermelon in Korea

    Directory of Open Access Journals (Sweden)

    Hae-Ryun Kwak

    2015-12-01

    Full Text Available Melon necrotic spot virus (MNSV was recently identified on watermelon (Citrullus vulgaris in Korea, displaying as large necrotic spots and vein necrosis on the leaves and stems. The average occurrence of MNSV on watermelon was found to be 30–65% in Hapcheon and Andong City, respectively. Four isolates of the virus (MNSV-HW, MNSV-AW, MNSV-YW, and MNSV-SW obtained from watermelon plants in different areas were non-pathogenic on ten general indicator plants, including Chenopodium quinoa, while they infected systemically six varieties of Cucurbitaceae. The virus particles purified by 10–40% sucrose density gradient centrifugation had a typical ultraviolet spectrum, with a minimum at 245 nm and a maximum at 260 nm. The morphology of the virus was spherical with a diameter of 28–30 nm. Virus particles were observed scattered throughout the cytoplasm of watermelon cells, but no crystals were detected. An ELISA was conducted using antiserum against MNSV-HW; the optimum concentrations of IgG and conjugated IgG for the assay were 1 μl/ml and a 1:8,000–1:10,000 dilutions, respectively. Antiserum against MNSV-HW could capture specifically both MNSV-MN from melon and MNSV-HW from watermelon by IC/RT-PCR, and they were effectively detected with the same specific primer to produce product of 1,172 bp. The dsRNA of MNSV-HW had the same profile (4.5, 1.8, and 1.6 kb as that of MNSV-MN from melon. The nucleotide sequence of the coat protein of MNSV-HW gave a different phylogenetic tree, having 17.2% difference in nucleotide sequence compared with MNSV isolates from melon.

  5. Caracterização morfofisiológica e identificação molecular de isolados de Guignardia citricarpa, agente patogênico da mancha preta dos citros = Morphophysiological characterization and molecular identification of isolates of Guignardia citricarpa, a pathogenic agent of the citrus black spot

    Directory of Open Access Journals (Sweden)

    Marilda Pereira Caixeta

    2008-12-01

    Full Text Available O presente trabalho teve como objetivo identificar 11 isolados de Guignardia citricarpa, agente causal da mancha preta dos citros (MPC, obtidos de frutas cítricas sintomáticas de diferentes regiões geográficas, por meio da PCR e caracterização morfofisiológica das estruturas propagativas, esporulação e crescimento micelial emdiferentes meios de cultura, temperaturas e regimes de luz, nas condições de laboratório. Pelo teste de PCR, todos os isolados foram identificados como o patógeno G. citricarpa. Os isolados caracterizados foram submetidos às temperaturas de 20, 25 e 30ºC, em regime de luz contínua, escuro contínuo e fotoperíodos de 12 horas, durante 24 dias. Utilizaram-se os meios de cultura aveia-ágar (AA, batata-dextrose-ágar (BDA e cenoura-dextrose-ágar (CDA. Os resultados mostraram que ocorreu interação entre os diferentes meios de cultura, temperaturas e fotoperíodos. O meio de cultura que melhor estimulou o crescimento micelial foi o CDA a 25ºC sob o fotoperíodo de 12h. A maior produção de esporos (conídios foi verificada no meio BDA a 20ºC, no fotoperíodo de 12 horas. No meio CDA, não ocorreu esporulação de nenhum isolado. Sob a temperatura de 30ºC, foiverificada apenas a produção de hifas e picnídios para a maioria dos isolados, em todos os meios de cultura e fotoperíodo testados.Thepresent work aims to identify 11 isolates of Guignardia citricarpa, the causal agent of the citrus black spot (CBS, obtained from affected fruit in different geographical regions, through PCR and morphophysiological characterization of propagative structures, sporing and mycelial growth in different means of culture, temperatures and photoperiods, under laboratory conditions. Through the PCR test, all isolates were identified as being the G.citricarpa pathogen. The characterized isolates were subjected to evaluations at temperatures of 20, 25 and 30ºC, in continuous light, continuous darkness, and alternating 12

  6. Dried blood spots, valid screening for viral hepatitis and human immunodeficiency virus in real-life

    DEFF Research Database (Denmark)

    Mössner, Belinda K; Staugaard, Benjamin; Jensen, Janne

    2016-01-01

    AIM: To detect chronic hepatitis B (CHB), chronic hepatitis C (CHC) and human immunodeficiency virus (HIV) infections in dried blood spot (DBS) and compare these samples to venous blood sampling in real-life. METHODS: We included prospective patients with known viral infections from drug treatment......, but correctly classified 95% of the anti-HCV-positive patients with chronic and past infections. Anti-HBc and anti-HBS showed low sensitivity in DBS (68% and 42%). CONCLUSION: DBS sampling, combined with an automated analysis system, is a feasible screening method to diagnose chronic viral hepatitis and HIV...

  7. Evaluation of peanut genotypes for resistance to Tomato spotted wilt virus by mechanical and thrips inoculation.

    OpenAIRE

    NASCIMENTO, L.C. do; PENSUK, V.; COSTA, N.P. da; ASSIS FILHO, F.M. de; PIO-RIBEIRO, G.; DEOM, C.M.; Sherwood, J.

    2006-01-01

    The objective of this work was to evaluate the reactions of three peanut breeding lines (IC-10, IC-34, and ICGV 86388) to Tomato spotted wilt virus (TSWV) by mechanical and thrips inoculation, under greenhouse conditions, and compare them to the reactions of cultivars SunOleic, Georgia Green, and the breeding line C11-2-39. TSWV infection by mechanical inoculation was visually assessed using an index ranging from 0 (no symptoms) to 4 (apical death). Enzyme-linked immunosorbent assay was used ...

  8. Analysis of the coding-complete genomic sequence of groundnut ringspot virus suggests a common ancestor with tomato chlorotic spot virus.

    Science.gov (United States)

    de Breuil, Soledad; Cañizares, Joaquín; Blanca, José Miguel; Bejerman, Nicolás; Trucco, Verónica; Giolitti, Fabián; Ziarsolo, Peio; Lenardon, Sergio

    2016-08-01

    Groundnut ringspot virus (GRSV) and tomato chlorotic spot virus (TCSV) share biological and serological properties, so their identification is carried out by molecular methods. Their genomes consist of three segmented RNAs: L, M and S. The finding of a reassortant between these two viruses may complicate correct virus identification and requires the characterization of the complete genome. Therefore, we present for the first time the complete sequences of all the genes encoded by a GRSV isolate. The high level of sequence similarity between GRSV and TCSV (over 90 % identity) observed in the genes and proteins encoded in the M RNA support previous results indicating that these viruses probably have a common ancestor.

  9. Melon necrotic spot virus Replication Occurs in Association with Altered Mitochondria.

    Science.gov (United States)

    Gómez-Aix, Cristina; García-García, María; Aranda, Miguel A; Sánchez-Pina, María Amelia

    2015-04-01

    Melon necrotic spot virus (MNSV) (genus Carmovirus, family Tombusviridae) is a single-stranded, positive-sense RNA virus that has become an experimental model for the analysis of cell-to-cell virus movement and translation of uncapped viral RNAs, whereas little is known about its replication. Analysis of the cytopathology after MNSV infection showed the specific presence of modified organelles that resemble mitochondria. Immunolocalization of the glycine decarboxylase complex (GDC) P protein in these organelles confirmed their mitochondrial origin. In situ hybridization and immunolocalization experiments showed the specific localization of positive-sense viral RNA, capsid protein (CP), and double-stranded (ds)RNA in these organelles meaning that replication of the virus takes place in association with them. The three-dimensional reconstructions of the altered mitochondria showed the presence of large, interconnected, internal dilations which appeared to be linked to the outside cytoplasmic environment through pores and/or complex structures, and with lipid bodies. Transient expression of MNSV p29 revealed that its specific target is mitochondria. Our data document the extensive reorganization of host mitochondria induced by MNSV, which provides a protected environment to viral replication, and show that the MNSV p29 protein is the primary determinant of this effect in the host.

  10. Resistance to tomato spotted wilt virus infection in transgenic tobacco expressing the viral nucleocapsid gene.

    Science.gov (United States)

    MacKenzie, D J; Ellis, P J

    1992-01-01

    A recombinant plasmid containing the entire tomato spotted with virus (TSWV) nucleocapsid gene, with the exception of nucleotide encoding three N-terminal amino acids, was isolated by screening a complementary DNA library, prepared against random primed viral RNA, using a specific monoclonal antibody. The insert contained in plasmid pTSW1 was repaired and amplified by polymerase chain reaction, and the complete nucleocapsid protein gene was introduced into Nicotiana tabacum 'Samsun' by leaf disk transformation using Agrobacterium tumefaciens. Transgenic plants expressing the viral nucleocapsid protein were resistant to subsequent infection following mechanical inoculation with TSWV as indicated by a lack of systemic symptoms and little or no systemic accumulation of virus as determined by double antibody sandwich enzyme-liked immunosorbent assay. These results further extend the applicability of coat protein-mediated resistance, as previously demonstrated for a number of simple plant viruses composed of a positive-sense RNA genome encapsidated with a single species of coat protein, to a membrane-encapsidated, multi-component, negative-sense RNA virus.

  11. An update on mechanism of entry of white spot syndrome virus into shrimps.

    Science.gov (United States)

    Verma, Arunima Kumar; Gupta, Shipra; Singh, Shivesh Pratap; Nagpure, Naresh Sahebrao

    2017-08-01

    Host-parasite relationships can be best understood at the level of protein-protein interaction between host and pathogen. Such interactions are instrumental in understanding the important stages of life cycle of pathogen such as adsorption of the pathogen on host surface followed by effective entry of pathogen into the host body, movement of the pathogen across the host cytoplasm to reach the host nucleus and replication of the pathogen within the host. White Spot Disease (WSD) is a havoc for shrimps and till date no effective treatment is available against the disease. Moreover information regarding the mechanism of entry of White Spot Syndrome Virus (WSSV) into shrimps, as well as knowledge about the protein interactions occurring between WSSV and shrimp during viral entry are still at very meagre stage. A cumulative and critically assessed information on various viral-shrimp interactions occurring during viral entry can help to understand the exact pathway of entry of WSSV into the shrimp which in turn can be used to device drugs that can stop the entry of virus into the host. In this context, we highlight various WSSV and shrimp proteins that play role in the entry mechanism along with the description of the interaction between host and pathogen proteins. Copyright © 2017 Elsevier Ltd. All rights reserved.

  12. Evaluation of Alternatives to Carbamate and Organophosphate Insecticides Against Thrips and Tomato Spotted Wilt Virus in Peanut Production.

    Science.gov (United States)

    Marasigan, K; Toews, M; Kemerait, R; Abney, M R; Culbreath, A; Srinivasan, R

    2016-04-01

    Thrips are important pests of peanut. They cause severe feeding injuries on peanut foliage in the early season. They also transmit Tomato spotted wilt virus (TSWV), which causes spotted wilt disease. At-plant insecticides and cultivars that exhibit field resistance to TSWV are often used to manage thrips and spotted wilt disease. Historically, peanut growers used the broad-spectrum insecticides aldicarb (IRAC class 1A; Temik) and phorate (IRAC class 1B; Thimet) for managing thrips and thereby reducing TSWV transmission. Aldicarb has not been produced since 2011 and its usage in peanut will be legally phased out in 2018; therefore, identification of alternative chemistries is critical for thrips and spotted wilt management. Here, eight alternative insecticides, with known thrips activity, were evaluated in field trials conducted from 2011 through 2013. In addition, different application methods of alternatives were also evaluated. Imidacloprid (Admire Pro), thiamethoxam (Actara), spinetoram (Radiant), and cyantraniliprole (Exirel) were as effective as aldicarb and phorate in suppressing thrips, but none of the insecticides significantly suppressed spotted wilt incidence. Nevertheless, greenhouse assays demonstrated that the same alternative insecticides were effective in suppressing thrips feeding and reducing TSWV transmission. Spotted wilt incidence in the greenhouse was more severe (∼80%) than in the field (5–25%). In general, field resistance to TSWV in cultivars only marginally influenced spotted wilt incidence. Results suggest that effective management of thrips using alternative insecticides and subsequent feeding reduction could improve yields under low to moderate virus pressure.

  13. Oxylipin biosynthesis genes positively regulate programmed cell death during compatible infections with the synergistic pair potato virus X-potato virus Y and Tomato spotted wilt virus.

    Science.gov (United States)

    García-Marcos, Alberto; Pacheco, Remedios; Manzano, Aranzazu; Aguilar, Emmanuel; Tenllado, Francisco

    2013-05-01

    One of the most severe symptoms caused by compatible plant-virus interactions is systemic necrosis, which shares common attributes with the hypersensitive response to incompatible pathogens. Although several studies have identified viral symptom determinants responsible for systemic necrosis, mechanistic models of how they contribute to necrosis in infected plants remain scarce. Here, we examined the involvement of different branches of the oxylipin biosynthesis pathway in the systemic necrosis response caused either by the synergistic interaction of Potato virus X with Potato virus Y (PVX-PVY) or by Tomato spotted wilt virus (TSWV) in Nicotiana benthamiana. Silencing either 9-lipoxygenase (LOX), 13-LOX, or α-dioxygenase-1 (α-DOX-1) attenuated the programmed cell death (PCD)-associated symptoms caused by infection with either PVX-PVY or TSWV. In contrast, silencing of the jasmonic acid perception gene, COI1 (Coronatine insensitive 1), expedited cell death during infection with compatible viruses. This correlated with an enhanced expression of oxylipin biosynthesis genes and dioxygenase activity in PVX-PVY-infected plants. Moreover, the Arabidopsis thaliana double lox1 α-dox-1 mutant became less susceptible to TSWV infection. We conclude that oxylipin metabolism is a critical component that positively regulates the process of PCD during compatible plant-virus interactions but does not play a role in restraining virus accumulation in planta.

  14. Complete nucleotide sequence of Pelargonium zonate spot virus and its relationship with the family Bromoviridae.

    Science.gov (United States)

    Finetti-Sialer, M; Gallitelli, D

    2003-11-01

    The complete sequence of the Pelargonium zonate spot virus (PZSV) genome was determined. It comprises 8477 nt, distributed in three positive-strand RNA species encoding four proteins. RNA-1 is 3383 nt long, with an ORF that encodes a polypeptide with a molecular mass of 108 419 Da (denoted protein 1a). This protein contains the conserved sequence motifs I-III of type I methyltransferases and the seven consensus motifs of the helicases of superfamily 1. RNA-2 is 2435 nt long and encodes a major polypeptide with a molecular mass of 78 944 Da (denoted protein 2a), which shows identity to the RNA-dependent RNA polymerases of positive-strand RNA viruses. RNA-3 is 2659 nt long and contains two major ORFs. The first ORF is located in the 5' portion of the genome and sequence comparison of the putative translation product revealed similarities with the 30K superfamily of virus movement proteins. The second ORF is located in the 3' half and encodes the viral coat protein, which is expressed via a subgenomic RNA, RNA-4. The transcription initiation site of RNA-4 maps to the intergenic region of RNA-3. The organization of the PZSV genome, including the primary structure of terminal non-coding regions, strongly suggests that this virus belongs to the family Bromoviridae. The overall biological and genomic characteristics of PZSV indicate affinities in diverging directions with one or other of the virus species in this family, thus enabling it to be considered as a possible representative of a new genus within the family Bromoviridae.

  15. Restricted Spread of Tomato spotted wilt virus in Thrips-Resistant Pepper.

    Science.gov (United States)

    Maris, P C; Joosten, N N; Goldbach, R W; Peters, D

    2003-10-01

    ABSTRACT Spread of Tomato spotted wilt virus (TSWV) and population development of its vector Frankliniella occidentalis were studied on the pepper accessions CPRO-1 and Pikante Reuzen, which are resistant and susceptible to thrips, respectively. Viruliferous thrips were released on plants of each accession (nonchoice tests) or on plants in a 1:1 mixture of both accessions (choice tests) in small cages containing 8 or 16 plants. Significantly fewer CPRO-1 plants became infected in the primary infection phase in both tests. In the nonchoice test, virus infection of the resistant plants did not increase after the initial infection, but all plants eventually became infected when mixtures of both cultivars were challenged in the secondary infection phase. Secondary spread of TSWV from an infected resistant or susceptible source plant was significantly slower to resistant plants than to susceptible plants, independent of source plant phenotype. The restricted introduction and spread of TSWV in the thrips-resistant cultivar was confirmed in a large-scale greenhouse experiment. The restricted and delayed TSWV spread to plants of the resistant accession in both the cage and the greenhouse experiment was explained by impeded thrips population development. The results obtained indicate that thrips resistance may provide a significant protection to TSWV infection, even when the crop is fully susceptible to the virus.

  16. Transcriptome Analysis of Litopenaeus vannamei in Response to White Spot Syndrome Virus Infection

    Science.gov (United States)

    Chen, Xiuli; Xie, Daxiang; Zhao, Yongzhen; Yang, Chunling; Li, Yongmei; Ma, Ning; Li, Ming; Yang, Qiong; Liao, Zhenping; Wang, Hui

    2013-01-01

    Pacific white shrimp (Litopenaeus vannamei) is the most extensively farmed crustacean species in the world. White spot syndrome virus (WSSV) is one of the major pathogens in the cultured shrimp. However, the molecular mechanisms of the host-virus interaction remain largely unknown. In this study, the impact of WSSV infection on host gene expression in the hepatopancreas of L. vannamei was investigated through the use of 454 pyrosequencing-based RNA-Seq of cDNA libraries developed from WSSV-challenged shrimp or normal controls. By comparing the two cDNA libraries, we show that 767 host genes are significantly up-regulated and 729 genes are significantly down-regulated by WSSV infection. KEGG analysis of the differentially expressed genes indicated that the distribution of gene pathways between the up- and down-regulated genes is quite different. Among the differentially expressed genes, several are found to be involved in various processes of animal defense against pathogens such as apoptosis, mitogen-activated protein kinase (MAPK) signaling, toll-like receptor (TLR) signaling, Wnt signaling and antigen processing and presentation pathways. The present study provides valuable information on differential expression of L. vannamei genes following WSSV infection and improves our current understanding of this host-virus interaction. In addition, the large number of transcripts obtained in this study provides a strong basis for future genomic research on shrimp. PMID:23991181

  17. Caracterização molecular do vírus da raiva isolado de Desmodus rotundus capturados no Estado do Rio de Janeiro Molecular characterization of rabies virus isolated from Desmodus rotundus captured in Rio de Janeiro State

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    L.F.P. Vieira

    2010-04-01

    Full Text Available Caracterizou-se filogeneticamente o vírus da raiva, isolado de morcegos hematógafos (Demodus rotundus. Cento e noventa e nove D. rotundus foram capturados em cinco abrigos, no Norte e Noroeste do Estado do Rio de Janeiro e sul do Espírito Santo. Sete deles foram positivos para a raiva. Amostras desses vírus foram sequenciadas e comparadas com sequências provenientes de diversos estados brasileiros. As sequências de vírus da raiva isoladas, na região norte do Estado do Rio de Janeiro, mostraram características que as distinguem de amostras de vírus isoladas em outras regiões do país, no entanto foram idênticas às isoladas de bovinos no noroeste do Rio de Janeiro.Rabies samples isolated from vampire bats captured in the Rio de Janeiro State were phylogenetically analyzed. One hundred and ninety nine vampire bats were captured from five shelters from North and Northwest of Rio de Janeiro and South of Espírito Santo States. Seven of them were positive for rabies. Theses samples were sequenced and compared with rabies virus sequences from several Brazilian states. The sequences of rabies virus, isolated in the present work, from North of Rio de Janeiro State, showed characteristics that differ of the sequences isolated from bats from other Brazilian regions. However, they were identical to samples isolated from cattle in Northwest of Rio de Janeiro state.

  18. Evolutionary analysis of tomato Sw-5 resistance-breaking isolates of Tomato spotted wilt virus.

    Science.gov (United States)

    López, Carmelo; Aramburu, José; Galipienso, Luis; Soler, Salvador; Nuez, Fernando; Rubio, Luis

    2011-01-01

    Tomato spotted wilt virus (TSWV) causes severe economic losses in many crops worldwide and often overcomes resistant cultivars used for disease control. Comparison of nucleotide and amino acid sequences suggested that tomato resistance conferred by the gene Sw-5 can be overcome by the amino acid substitution C to Y at position 118 (C118Y) or T120N in the TSWV movement protein, NSm. Phylogenetic analysis revealed that substitution C118Y has occurred independently three times in the studied isolates by convergent evolution, whereas the substitution T120N was a unique event. Analysis of rates of non-synonymous and synonymous changes at individual codons showed that substitution C118Y was positively selected.

  19. Kualitas Lingkungan Tambak Insentif Litapenaeus Vannamei Dalam Kaitannya Dengan Prevalensi Penyakit White Spot Syndrome Virus

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    Yunita Maimunah

    2015-04-01

    Full Text Available Increasing number of vannamei shrimp (Litapenaeus vannamei ponds are switching from traditional to intensive farming systems, the more impact resulting among other potential environmental pollution. Pollution of the environment can directly degrade water quality cultivation and facilitate access of pathogens to infect the host. In this study examines how the quality of the environment, population and genetic characteristics of shrimp that live in some intensive pond associated with a disease that often affects farmed shrimp is White Spot Syndrome Virus (WSSV. Acquisition and primary data collection is done by conducting interviews and direct observation in the measurement of water quality parameters of both physics and chemistry and morphology observation of shrimp as well as the ICP11 gene expression detection of WSSV disease in vannamei shrimp DNA in the laboratory.

  20. Binding of white spot syndrome virus to Artemia sp. cell membranes.

    Science.gov (United States)

    Feng, Shuying; Li, Guangda; Feng, Wenpo; Huang, Jie

    2013-10-01

    Using differential velocity centrifugation, cell membranes of Artemia sp. were prepared, and their binding to white spot syndrome virus (WSSV) was analyzed in vitro. The results indicated that WSSV can specifically bind to Artemia cell membranes, and that WSSV receptor very likely existed in this membrane, which suggested that Artemia sp. may be a reservoir of WSSV. This study investigated the specific WSSV binding site by performing competitive inhibition experiments using shrimp gill cell membranes to bind WSSV to Artemia cell membranes. The results showed that shrimp gill cell membranes had a distinct inhibition effect on the specific binding of Artemia cell membranes to WSSV. Thus, potentially similar WSSV receptors or binding sites existed on Artemia sp. cell membranes and shrimp gill cell membranes. Taken together, these findings may provide experimental basis for the development of an effective approach to controlling WSSV, and theoretical basis for the study of WSSV receptors. Copyright © 2013 Elsevier B.V. All rights reserved.

  1. Blue crabs Callinectes sapidus as potential biological reservoirs for white spot syndrome virus (WSSV).

    Science.gov (United States)

    Powell, James W B; Browdy, Craig L; Burge, Erin J

    2015-03-09

    White spot syndrome virus (WSSV) is a virulent pathogen of cultured shrimp and was first detected in farms in South Carolina (USA) in 1997 and subsequently in wild shrimp in 1999. We screened groups of 1808 wild Atlantic white shrimp Litopenaeus setiferus and 300 blue crabs Callinectes sapidus collected from South Carolina, Georgia, and Florida for the presence of WSSV using the Shrimple® immunoassay-strip test, with all positives and random subsets of negatives tested by TaqMan real-time PCR and in infectivity bioassays. Of 87 shrimp and 11 crabs that tested positive using the Shrimple® test, only a single C. sapidus was confirmed to be infected with WSSV by PCR and the infectivity bioassay. The data indicate that the prevalence of WSSV in these species is low in these southeastern US regions, but that C. sapidus may serve as a biological reservoir.

  2. First report of tomato chlorotic spot virus in sweet basil (Ocimum basilicum) and purslane (Portulaca oleracea) in Florida

    Science.gov (United States)

    Tomato chlorotic spot virus (TCSV) has been recently detected in tomato, pepper, hoya and vinca in Florida. Observations of additional crops in 2016 and 2017 revealed TCSV-like symptoms. Testing of these symptomatic plants identified three new hosts of TCSV in Florida: sweet basil (Ocimum basilicu...

  3. Iris Yellow Spot Virus in the Netherlands: Occurence in Onion and Confirmation of Transmission by Thrips tabaci

    NARCIS (Netherlands)

    Hoedjes, K.; Verhoeven, J.Th.J.; Goldbach, R.W.; Peters, D.

    2011-01-01

    Since its first detection in the Netherlands in 1992, Iris yellow spot virus (IYSV, genus Tospovirus) has been reported worldwide in Allium crops, in a few ornamentals and in a small number of weeds. After recent findings of IYSV in Alstroemeria and Eustoma in the Netherlands, a number of

  4. The use of fluorescence microscopy to visualise homotypic interactions of tomato spotted wilt virus nucleocapsid protein in living cells

    NARCIS (Netherlands)

    Snippe, M.; Borst, J.W.; Goldbach, R.W.; Kormelink, R.J.M.

    2005-01-01

    Fluorescence resonance energy transfer (FRET) and fluorescence lifetime imaging microscopy (FLIM) were employed to study homotypic protein¿protein interactions in living cells. To this end, the nucleocapsid (N) protein of tomato spotted wilt virus (TSWV) was expressed as a fusion protein with either

  5. Identification of Stressors that Affect White Spot Syndrome Virus (WSSV) Infection and Outbreak in Pond Cultured Penaeus monodon

    NARCIS (Netherlands)

    Tendencia Alapide, E.; Verreth, J.A.J.

    2011-01-01

    White spot syndrome virus (WSSV) has been a big problem to the worldwide shrimp industry. Exposure to stressors related to physicochemical water parameters affect WSSV infection but not all WSSV infections result in outbreaks. This paper describes a detailed monitoring of important physicochemical

  6. Symptom and Resistance of Cultivated and Wild Capsicum Accessions to Tomato Spotted Wilt Virus

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    Jung-Heon Han

    2011-04-01

    Full Text Available One hundred Capsicum accessions were screened for symptomatic response and resistance to Tomato spotted wilt virus-pb1 (TSWV-pb1. Symptom and its severity rating were checked by visual observation at 9, 12, 14, and 45 days after inoculation, respectively. Enzyme-linked immune-sorbent assay was performed all tested individuals on non-inoculated upper leaves after the third rating to indentify viral infection. Leaf curling was predominant in almost susceptible individuals of each accession. Stem necrosis was most frequent in wild species while yellowing in commercial hybrids and Korean land race cultivars. Ring spot, a typical symptom of TSWV, was rarely detected in some of a few accessions. Different levels of resistance to TSWV-pb1 were observed among the tested accessions. High level of resistance was detected in 4 commercial cultivars of Kpc- 35, -36, -57, and -62, and 8 wild species of PBI-11, C00105, PBC076, PBC280, PBC426, PBC495, PBC537, and PI201238 through seedling test by mechanical inoculation.

  7. Analysis of the Tomato spotted wilt virus ambisense S RNA-encoded hairpin structure in translation.

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    Christina Geerts-Dimitriadou

    Full Text Available BACKGROUND: The intergenic region (IR of ambisense RNA segments from animal- and plant-infecting (-RNA viruses functions as a bidirectional transcription terminator. The IR sequence of the Tomato spotted wilt virus (TSWV ambisense S RNA contains stretches that are highly rich in A-residues and U-residues and is predicted to fold into a stable hairpin structure. The presence of this hairpin structure sequence in the 3' untranslated region (UTR of TSWV mRNAs implies a possible role in translation. METHODOLOGY/PRINCIPAL FINDINGS: To analyse the role of the predicted hairpin structure in translation, various Renilla luciferase constructs containing modified 3' and/or 5' UTR sequences of the TSWV S RNA encoded nucleocapsid (N gene were analyzed for expression. While good luciferase expression levels were obtained from constructs containing the 5' UTR and the 3' UTR, luciferase expression was lost when the hairpin structure sequence was removed from the 3' UTR. Constructs that only lacked the 5' UTR, still rendered good expression levels. When in addition the entire 3' UTR was exchanged for that of the S RNA encoded non-structural (NSs gene transcript, containing the complementary hairpin folding sequence, the loss of luciferase expression could only be recovered by providing the 5' UTR sequence of the NSs transcript. Luciferase activity remained unaltered when the hairpin structure sequence was swapped for the analogous one from Tomato yellow ring virus, another distinct tospovirus. The addition of N and NSs proteins further increased luciferase expression levels from hairpin structure containing constructs. CONCLUSIONS/SIGNIFICANCE: The results suggest a role for the predicted hairpin structure in translation in concert with the viral N and NSs proteins. The presence of stretches highly rich in A-residues does not rule out a concerted action with a poly(A-tail-binding protein. A common transcription termination and translation strategy for plant

  8. Experimental transmission of white spot syndrome virus (WSSV) from crabs to shrimp Penaeus monodon.

    Science.gov (United States)

    Kanchanaphum, P; Wongteerasupaya, C; Sitidilokratana, N; Boonsaeng, V; Panyim, S; Tassanakajon, A; Withyachumnarnkul, B; Flegel, T W

    1998-09-11

    White spot syndrome virus (WSSV) of the black tiger prawn Penaeus monodon is a recently discovered baculo-like virus disease which is currently the cause of very serious and widespread losses in the shrimp industry in Thailand and elsewhere in Asia. Three suspected crab carriers of this virus commonly found in shrimp-rearing areas were investigated. These were Sesarma sp., Scylla serrata and Uca pugilator. All these crabs could be infected with WSSV by injection and they sustained heavy viral infections for up to 45 d (confirmed by normal histology, specific in situ DNA hybridization and PCR amplification) without visible signs of disease or mortality. All of them also transferred the disease to P. monodon via water while physically separated in aquarium cohabitation tests. Transfer of the virus to the shrimp was monitored using in situ DNA hybridization and PCR assay at 12 h intervals after cohabitation began. With U. pugilator, WSSV could be detected in the shrimp cohabitants after 24 h using PCR amplification and after 60 h using in situ hybridization. With S. serrata, the shrimp were positive for WSSV after 36 h using PCR and after 60 h using DNA in situ hybridization. With Sesarma sp. they were positive after 48 h using PCR and 72 h using in situ hybridization. These laboratory studies demonstrated that crab carriers of WSSV may pose a real threat to cultivated shrimp. However, the studies were carried out in containers with a small volume and with relatively clean sea water as compared to shrimp cultivation ponds. Pond-based studies are now needed to determine whether factors such as pond volume, pond water quality and shrimp and crab behavior can influence the rate and success of transfer.

  9. Genetic grouping of avian infectious bronchitis virus isolated in Brazil based on RT-PCR/RFLP analysis of the S1 gene Agrupamento genético de isolados do vírus da bronquite infecciosa das aves no Brasil com base na análise do gene S1 por RT-PCR-RFLP

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    Maria de Fátima S. Montassier

    2008-03-01

    Full Text Available Twelve Brazilian isolates and one reference vaccine strain of avian infectious bronchitis virus (IBV were propagated in embryonating chicken eggs. The entire S1 glycoprotein gene of these viruses was analysed by reverse-transcriptase-polymerase chain reaction and restriction fragment length polymorphism (RT-PCR-RFLP, using the restriction enzymes HaeIII, XcmI and BstyI. The RFLP patterns led to the classification of these isolates into five distinct genotypes: A, B, C, D and Massachusetts. Five of twelve isolates were grouped in Massachusetts genotype and the remaining seven viruses were classified into four distinct genotypes: A (2, B (2, C (2 or D (1. Such genotyping classification agreed with previous immunological analysis for most of these viruses, highlighting the occurrence of a relevant variability among the IBV strains that are circulating in Brazilian commercial poultry flocks.Doze isolados de campo do Brasil e uma estirpe de referência vacinal do vírus da bronquite infecciosa das aves (VBI foram propagadas em ovos embrionados SPF. O gene S1 dessas amostras foi analisado por RT-PCR seguido de RFLP, empregando-se as enzimas de restrição HaeIII, XcmI e BstyI. Observou-se a existência de cinco genotipos diferentes: M (Massachusetts, A , B, C e D. Cinco dos doze isolados de campo do VBI foram classificados no genótipo Massachusetts e os sete vírus restantes foram classificados em quatro genotipos diferentes; A (2, B (2, C (2 ou D (1. Os resultados desta genotipagem concordam com os dados obtidos na análise imunológica previamente realizada para a maior parte destes vírus, destacando a ocorrência de uma variabilidade marcante entre os isolados do VBI que estão circulando nas granjas avícolas comerciais do Brasil.

  10. White spot syndrome virus infection: Threat to crustacean biodiversity in Vembanad Lake, India

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    Toms C. Joseph

    2015-09-01

    Full Text Available The Vembanad Lake located on the south-west coast of India, an ecological hotspot is the nursing ground of many economically important crustaceans. The prevalence of white spot syndrome virus (WSSV among crustaceans from farmed, estuarine and marine environments surrounding the Vembanad Lake, India was detected using PCR. A total of 308 samples from aquaculture ponds consisting of six species of crustaceans collected from five different farms were tested for the presence of WSSV. Of these, 67% were found to carry the virus. A total of 258 samples of crustaceans from the Cochin backwater system that forms a part of the Vembanad lake viz., Metapenaeus dobsoni, Metapenaeus monoceros, Penaeus monodon and Penaeus indicus were found to contain WSSV in 62% of the samples. Fifteen species of crustaceans caught from the seas off Cochin were also screened for the presence of WSSV. Out of these, twelve species had WSSV incidence levels ranging from 6–23%. WSSV was not detected from three species of deep sea crustaceans tested. The black tiger shrimp, Penaeus monodon had the highest incidence of WSSV among the species screened in farmed, estuarine and marine environments.

  11. OIE white spot syndrome virus PCR gives false-positive results in Cherax quadricarinatus.

    Science.gov (United States)

    Claydon, Kerry; Cullen, Bradford; Owens, Leigh

    2004-12-13

    White spot syndrome virus (WSSV) is an intranuclear bacilliform virus (IBV) that is a serious, notifiable crustacean pathogen. The Office International des Epizooties (OIE) PCR protocol for WSSV uses primer sets initially developed by Lo et al. (1996). It yields a first-step PCR amplicon of 1441 bp and a nested PCR amplicon of 941 bp. An amplicon (941 bp) purported to specifically detect WSSV was obtained when using template DNA extracted from Cherax quadricarinatus in a WSSV PCR detection protocol recommended by the OIE. Sequencing and analysis of the 941 bp amplicon and an occasional 550 bp amplicon from C. quadricarinatus revealed no phylogenetic relationship with WSSV, and suggested a possible lack of sufficient primer specificity for WSSV in the OIE test. This suggestion was supported by the fact that the OIE outer primer sequence (146F1) was present in both the forward and reverse position of the 941 bp and the forward position of the 550 bp nested amplicons from C. quadricarinatus. As WSSV is a notifiable pathogen, the consequences of false-positive results are harsh in WSSV-free zones and can lead to incorrect quarantine and unnecessary destruction of animals. Therefore, urgent attention and revision is necessary for the current OIE PCR protocol for WSSV detection.

  12. Expression and characterization of a soluble form of tomato spotted wilt virus glycoprotein GN.

    Science.gov (United States)

    Whitfield, Anna E; Ullman, Diane E; German, Thomas L

    2004-12-01

    Tomato spotted wilt virus (TSWV), a member of the Tospovirus genus within the Bunyaviridae, is an economically important plant pathogen with a worldwide distribution. TSWV is transmitted to plants via thrips (Thysanoptera: Thripidae), which transmit the virus in a persistent propagative manner. The envelope glycoproteins, G(N) and G(C), are critical for the infection of thrips, but they are not required for the initial infection of plants. Thus, it is assumed that the envelope glycoproteins play important roles in the entry of TSWV into the insect midgut, the first site of infection. To directly test the hypothesis that G(N) plays a role in TSWV acquisition by thrips, we expressed and purified a soluble, recombinant form of the G(N) protein (G(N)-S). The expression of G(N)-S allowed us to examine the function of G(N) in the absence of other viral proteins. We detected specific binding to thrips midguts when purified G(N)-S was fed to thrips in an in vivo binding assay. The TSWV nucleocapsid protein and human cytomegalovirus glycoprotein B did not bind to thrips midguts, indicating that the G(N)-S-thrips midgut interaction is specific. TSWV acquisition inhibition assays revealed that thrips that were concomitantly fed purified TSWV and G(N)-S had reduced amounts of virus in their midguts compared to thrips that were fed TSWV only. Our findings that G(N)-S binds to larval thrips guts and decreases TSWV acquisition provide evidence that G(N) may serve as a viral ligand that mediates the attachment of TSWV to receptors displayed on the epithelial cells of the thrips midgut.

  13. Molecular characterization of a divergent strain of calla lily chlorotic spot virus infecting celtuce (Lactuca sativa var. augustana) in China.

    Science.gov (United States)

    Wu, Xiaodong; Wu, Xiaoyun; Li, Wenbin; Cheng, Xiaofei

    2018-02-01

    Through sequencing and assembly of small RNAs, an orthotospovirus was identified from a celtuce plant (Lactuca sativa var. augustana) showing vein clearing and chlorotic spots in the Zhejiang province of China. The S, M, and L RNAs of this orthotospovirus were determined to be 3146, 4734, and 8934 nt, respectively, and shared 30.4-72.5%, 43.4-80.8%, and 29.84-82.9% nucleotide sequence identities with that of known orthotospoviruses. The full length nucleoprotein (N) of this orthotospovirus shared highest amino acid sequence identity (90.25%) with that of calla lily chlorotic spot virus isolated from calla lily (CCSV-calla) [China: Taiwan: 2001] and tobacco (CCSV-LJ1) [China: Lijiang: 2014]. Phylogenetic analyses showed that this orthotospovirus is phylogenetically associated with CCSV isolates and clustered with CCSV, tomato zonate spot virus (TZSV), and tomato necrotic spot-associated virus (TNSaV) in a separate sub-branch. These results suggest that this orthotospovirus is a divergent isolate of CCSV and was thus named CCSV-Cel [China: Zhejiang: 2017].

  14. Tomato chocolate spot virus, a member of a new torradovirus species that causes a necrosis-associated disease of tomato in Guatemala.

    Science.gov (United States)

    Batuman, O; Kuo, Y-W; Palmieri, M; Rojas, M R; Gilbertson, R L

    2010-06-01

    Tomatoes in Guatemala have been affected by a new disease, locally known as "mancha de chocolate" (chocolate spot). The disease is characterized by distinct necrotic spots on leaves, stems and petioles that eventually expand and cause a dieback of apical tissues. Samples from symptomatic plants tested negative for infection by tomato spotted wilt virus, tobacco streak virus, tobacco etch virus and other known tomato-infecting viruses. A virus-like agent was sap-transmitted from diseased tissue to Nicotiana benthamiana and, when graft-transmitted to tomato, this agent induced chocolate spot symptoms. This virus-like agent also was sap-transmitted to Datura stramonium and Nicotiana glutinosa, but not to a range of non-solanaceous indicator plants. Icosahedral virions approximately 28-30 nm in diameter were purified from symptomatic N. benthamiana plants. When rub-inoculated onto leaves of N. benthamiana plants, these virions induced symptoms indistinguishable from those in N. benthamiana plants infected with the sap-transmissible virus associated with chocolate spot disease. Tomatoes inoculated with sap or grafted with shoots from N. benthamiana plants infected with purified virions developed typical chocolate spot symptoms, consistent with this virus being the causal agent of the disease. Analysis of nucleic acids associated with purified virions of the chocolate-spot-associated virus, revealed a genome composed of two single-stranded RNAs of approximately 7.5 and approximately 5.1 kb. Sequence analysis of these RNAs revealed a genome organization similar to recently described torradoviruses, a new group of picorna-like viruses causing necrosis-associated diseases of tomatoes in Europe [tomato torrado virus (ToTV)] and Mexico [tomato apex necrosis virus (ToANV) and tomato marchitez virus (ToMarV)]. Thus, the approximately 7.5 kb and approximately 5.1 kb RNAs of the chocolate-spot-associated virus corresponded to the torradovirus RNA1 and RNA2, respectively; however

  15. Tomato chocolate spot virus, a member of a new torradovirus species that causes a necrosis-associated disease of tomato in Guatemala

    Science.gov (United States)

    Batuman, O.; Kuo, Y.-W.; Palmieri, M.; Rojas, M. R.

    2010-01-01

    Tomatoes in Guatemala have been affected by a new disease, locally known as “mancha de chocolate” (chocolate spot). The disease is characterized by distinct necrotic spots on leaves, stems and petioles that eventually expand and cause a dieback of apical tissues. Samples from symptomatic plants tested negative for infection by tomato spotted wilt virus, tobacco streak virus, tobacco etch virus and other known tomato-infecting viruses. A virus-like agent was sap-transmitted from diseased tissue to Nicotiana benthamiana and, when graft-transmitted to tomato, this agent induced chocolate spot symptoms. This virus-like agent also was sap-transmitted to Datura stramonium and Nicotiana glutinosa, but not to a range of non-solanaceous indicator plants. Icosahedral virions ~28–30 nm in diameter were purified from symptomatic N. benthamiana plants. When rub-inoculated onto leaves of N. benthamiana plants, these virions induced symptoms indistinguishable from those in N. benthamiana plants infected with the sap-transmissible virus associated with chocolate spot disease. Tomatoes inoculated with sap or grafted with shoots from N. benthamiana plants infected with purified virions developed typical chocolate spot symptoms, consistent with this virus being the causal agent of the disease. Analysis of nucleic acids associated with purified virions of the chocolate-spot-associated virus, revealed a genome composed of two single-stranded RNAs of ~7.5 and ~5.1 kb. Sequence analysis of these RNAs revealed a genome organization similar to recently described torradoviruses, a new group of picorna-like viruses causing necrosis-associated diseases of tomatoes in Europe [tomato torrado virus (ToTV)] and Mexico [tomato apex necrosis virus (ToANV) and tomato marchitez virus (ToMarV)]. Thus, the ~7.5 kb and ~5.1 kb RNAs of the chocolate-spot-associated virus corresponded to the torradovirus RNA1 and RNA2, respectively; however, sequence comparisons revealed 64–83% identities with

  16. Three decades of managing Tomato spotted wilt virus in peanut in southeastern United States.

    Science.gov (United States)

    Srinivasan, R; Abney, M R; Culbreath, A K; Kemerait, R C; Tubbs, R S; Monfort, W S; Pappu, H R

    2017-09-15

    Southeastern states namely Georgia, Florida, and Alabama produce two-thirds of the peanuts in the United States. Thrips-transmitted Tomato spotted wilt virus (TSWV), which causes spotted wilt disease, has been a major impediment to peanut production for the past three decades. The cultivars grown in the 1980s were extremely susceptible to TSWV. Early yield losses extended to tens of millions of dollars each year (up to 100% loss in many fields). This situation led to the creation of an interdisciplinary team known as "SWAT: Spotted Wilt Action Team". Initial efforts focused on risk mitigation using a combination of chemical and cultural management practices along with a strong investment in breeding programs. Beginning in the mid 1990s, cultivars with field resistance were developed and integrated with cultural and chemical management options. A Risk Mitigation Index (Peanut Rx) was made available to growers to assess risks, and provide options for mitigating risks such as planting field resistant cultivars with in-furrow insecticides, planting after peak thrips incidence, planting in twin rows, and increasing seeding rates. These efforts helped curtail losses due to spotted wilt. The Peanut Rx continues to be refined every year based on new research findings. Breeding efforts, predominantly in Georgia and Florida, continue to develop cultivars with incremental field resistance. The present-day cultivars (third-generation TSWV-resistant cultivars released after 2010) possess substantially greater field resistance than second-generation (cultivars released from 2000 to 2010) and first-generation (cultivars released from 1994 to 2000) TSWV resistant cultivars. Despite increased field resistance, these cultivars are not immune to TSWV and succumb under high thrips and TSWV pressure. Therefore, field resistant cultivars cannot serve as a 'stand-alone' option and have to be integrated with other management options. The mechanism of resistance is also unknown in field

  17. Detection of Tomato spotted wilt virus in its vector Frankliniella occidentalis by reverse transcription-polymerase chain reaction.

    Science.gov (United States)

    Mason, Giovanna; Roggero, Piero; Tavella, Luciana

    2003-04-01

    A method for rapid and reliable detection of Tomato spotted wilt virus (TSWV) (Tospovirus, Bunyaviridae) in its vector Frankliniella occidentalis (Thysanoptera Thripidae) would be a useful tool for studying the epidemiology of this virus. A RT-PCR method developed for this purpose is reported. The method was tested on thrips involved in laboratory transmission trials and on thrips collected in the field, whose capability to transmit TSWV was checked previously by leaf disk assays. The RT-PCR results were consistent with the results obtained by the leaf disk assays. Among thrips involved in laboratory experiments, 97% of the adults that transmitted TSWV were positive by RT-PCR; as did some non-transmitter adults reacted, whereas among field-collected thrips only the individuals able to transmit were positive by RT-PCR. In addition, healthy thrips were allowed to feed as adults on virus-infected leaves for 48 h, and then examined by RT-PCR immediately or after starving or feeding on virus-free plants for various times, to determine if virus ingested (but not transmissible) was also detectable. The virus was detectable immediately after the feed or within 12 and 24 h for individuals starved or fed on virus-free plants, respectively, but not after those periods. Thus, the method could detect rapidly and reliably the virus in vectors from the field, providing 24 h of starving to avoid positive RT-PCR results from thrips simply carrying the virus.

  18. Determination of whether tomato spotted wilt virus replicates in Toxorhynchites amboinensis mosquitoes and the relatedness of this virus to phleboviruses (family Bunyaviridae).

    Science.gov (United States)

    Wang, M; Mitchell, C J; Hu, J S; Gonsalves, D; Calisher, C H

    1992-01-01

    Tomato spotted wilt virus (TSWV) has been reported to be morphologically, molecularly and structurally similar to viruses in the family Bunyaviridae. By various types of enzyme-linked immunosorbent assays (ELISA) and Western blot hybridizations, we tested TSWV with antibodies to 12 viruses in the Phlebovirus genus of this family. Serological relatedness was not found between TSWV and phleboviruses. However, one preparation of antibody to Arumowot virus reacted with a 53-kD protein from healthy plant extracts. Six-day-old adult Toxorhynchites amboinensis mosquitoes were inoculated with purified TSWV. Infectious virus was not detected in any of the injected insects during the 5-week test period. However, TSWV antigens were detected in these mosquitoes by ELISA at the original injected level for at least a week after injection. TSWV antigen concentration began to decrease thereafter, but remained at detectable levels for as long as 5 weeks after injection. However, there was no evidence that TSWV replicated in mosquitoes.

  19. Iris yellow spot virus (Tospovirus: Bunyaviridae): from obscurity to research priority.

    Science.gov (United States)

    Bag, Sudeep; Schwartz, Howard F; Cramer, Christopher S; Havey, Michael J; Pappu, Hanu R

    2015-04-01

    Iris yellow spot virus (IYSV) is in the genus Tospovirus, family Bunyaviridae, with a single-stranded, tri-segmented RNA genome with an ambisense genome organization. Members of the other genera in the family infect predominantly vertebrates and insects. IYSV is present in most Allium-growing regions of the world. Virions are pleomorphic particles of 80-120 nm in size. The particle consists of RNA, protein, glycoprotein and lipids. IYSV shares the genomic features of other tospoviruses: a segmented RNA genome of three RNAs, referred to as large (L), medium (M) and small (S). The L RNA codes for the RNA-dependent RNA polymerase (RdRp) in negative sense. The M RNA uses an ambisense coding strategy and codes for the precursor for the GN /GC glycoprotein in the viral complementary (vc) sense and a non-structural protein (NSm) in the viral (v) sense. The S RNA also uses an ambisense coding strategy with the coat protein (N) in vc sense and a non-structural protein (NSs) in the v sense. The virus is transmitted by Thrips tabaci Lindeman (Order: Thysanoptera; Family: Thripidae; onion thrips) and with less efficiency by Frankliniella fusca Hinds (tobacco thrips). HOST: IYSV has a relatively broad host range, including cultivated and wild onions, garlic, chives, leeks and several ornamentals. Some weeds are naturally infected by IYSV and may serve as alternative hosts for the virus. IYSV symptoms in Allium spp. are yellow- to straw-coloured, diamond-shaped lesions on leaves and flowering scapes. Diamond-shaped lesions are particularly pronounced on scapes. As the disease progresses, the lesions coalesce, leading to lodging of the scapes. In seed crops, this could lead to a reduction in yield and quality. Early to mid-season infection in bulb crops results in reduced vigour and bulb size. Resistant varieties are not available, but a limited number of accessions with field tolerance have been identified. Integrated disease management tactics, including sanitation, crop

  20. Identification of major QTLs underlying tomato spotted wilt virus resistance in peanut cultivar Florida-EP(TM) '113'.

    Science.gov (United States)

    Tseng, Yu-Chien; Tillman, Barry L; Peng, Ze; Wang, Jianping

    2016-09-06

    Spotted wilt caused by tomato spotted wilt virus (TSWV) is one of the major peanut (Arachis hypogaea L.) diseases in the southeastern United States. Occurrence, severity, and symptoms of spotted wilt disease are highly variable from season to season, making it difficult to efficiently evaluate breeding populations for resistance. Molecular markers linked to spotted wilt resistance could overcome this problem and allow selection of resistant lines regardless of environmental conditions. Florida-EP(TM) '113' is a spotted wilt resistant cultivar with a significantly lower infection frequency. However, the genetic basis is still unknown. The objective of this study is to map the major quantitative trait loci (QTLs) linked to spotted wilt resistance in Florida-EP(TM) '113'. Among 2,431 SSR markers located across the whole peanut genome screened between the two parental lines, 329 were polymorphic. Those polymorphic markers were used to further genotype a representative set of individuals in a segregating population. Only polymorphic markers on chromosome A01 showed co-segregation between genotype and phenotype. Genotyping by sequencing (GBS) of the representative set of individuals in the segregating population also depicted a strong association between several SNPs on chromosome A01 and the trait, indicating a major QTL on chromosome A01. Therefore marker density was enriched on the A01 chromosome. A linkage map with 23 makers on chromosome A01 was constructed, showing collinearity with the physical map. Combined with phenotypic data, a major QTL flanked by marker AHGS4584 and GM672 was identified on chromosome A01, with up to 22.7 % PVE and 9.0 LOD value. A major QTL controlling the spotted wilt resistance in Florida-EP(TM) '113' was identified. The resistance is most likely contributed by PI 576638, a hirsuta botanical-type line, introduced from Mexico with spotted wilt resistance. The flanking markers of this QTL can be used for further fine mapping and marker

  1. The nucleocapsid protein of an enveloped plant virus, Tomato spotted wilt virus, facilitates long-distance movement of Tobacco mosaic virus hybrids.

    Science.gov (United States)

    Zhang, Yongqiang; Zhang, Chao; Li, Weimin

    2012-01-01

    To investigate the potential role(s) of the nucleocapsid (N) protein of Tomato spotted wilt virus (TSWV), the open reading frame for the N protein was expressed from a Tobacco mosaic virus (TMV)-based vector encoding only the TMV replicase proteins. In the absence of other TSWV-encoded proteins, the transiently expressed N protein facilitated long-distance movement of the TMV-based hybrids in transgenic Nicotiana benthamiana [NB-MP(+)] expressing movement protein of TMV, thus providing the functional demonstration of the N protein in long-distance RNA movement. Removal of the N-terminal 39 amino acids (N-NΔ39), the C-terminal 26 amino acids (N-CΔ26) or both of them (N-NΔ39CΔ26) abolished the long-distance movement function, indicating the essential role of both N- and C-terminus. In contrast, alanine substitution of the phenylalanines at positions 242 and 246 (N242/262A), two crucial amino acids for homotypic interaction of the N protein, had little effect, suggesting that the N protein could function in long-distance movement in the form of monomers. In addition, both the wild type N and the alanine mutant N242/262A hardly induced local symptoms in NB-MP(+) plants and TMV-MP transgenic N. tabacum cv. Xanthi. The deletion mutants N-NΔ39, N-CΔ26 and N-NΔ39CΔ26, however, induced apparent symptoms of necrotic ringspots, necrosis or chlorotic spots in all inoculated leaves. On the basis of these findings, the potential role of N during the TSWV infection was discussed. To our knowledge, this is the first report that the N protein of an enveloped plant virus functioned in long-distance movement. Copyright © 2011 Elsevier B.V. All rights reserved.

  2. Toll receptor response to white spot syndrome virus challenge in giant freshwater prawns (Macrobrachium rosenbergii).

    Science.gov (United States)

    Feng, Jinling; Zhao, Lingling; Jin, Min; Li, Tingting; Wu, Lei; Chen, Yihong; Ren, Qian

    2016-10-01

    Toll receptors are evolutionary ancient families of pattern recognition receptors with crucial roles in invertebrate innate immune response. In this study, we identified a Toll receptor (MrToll) from giant freshwater prawns (Macrobrachium rosenbergii). The full-length cDNA of MrToll is 4257 bp, which encodes a putative protein of 1367 amino acids. MrToll contains 17 LRR domains, a transmembrane domain, and a TIR domain. Phylogenetic analysis showed that MrToll was grouped with Drosophila Toll7 and other arthropod Tolls. The transcripts of MrToll are mainly distributed in the heart, hepatopancreas, gills, stomach, and intestine. A low level of MrToll expression can be detected in hemocytes and the lymphoid organ. MrToll expression in gills was gradually upregulated to the highest level from 24 h to 48 h during the white spot syndrome virus (WSSV) challenge. The expression levels of the crustin (Cru) genes Cru3 and Cru7 in gills were relatively lower than those of Cru2 and Cru4. The expression levels of Cru3 and Cru7 were inhibited after the RNA interference of MrToll in gills during the WSSV challenge. The anti-lipopolysaccharide factor (ALF) genes ALF2, ALF3, ALF4, and ALF5 were also regulated by MrToll in gills during the virus challenge. These findings suggest that MrToll may contribute to the innate immune defense of M. rosenbergii against WSSV. Copyright © 2016 Elsevier Ltd. All rights reserved.

  3. A transcriptome study on Macrobrachium rosenbergii hepatopancreas experimentally challenged with white spot syndrome virus (WSSV).

    Science.gov (United States)

    Rao, Rama; Bhassu, Subha; Bing, Robin Zhu Ya; Alinejad, Tahereh; Hassan, Sharifah Syed; Wang, Jun

    2016-05-01

    The world production of shrimp such as the Malaysian giant freshwater prawn, Macrobrachium rosenbergii is seriously affected by the white spot syndrome virus (WSSV). There is an urgent need to understand the host pathogen interaction between M. rosenbergii and WSSV which will be able to provide a solution in controlling the spread of this infectious disease and lastly save the aquaculture industry. Now, using Next Generation Sequencing (NGS), we will be able to capture the response of the M. rosenbergii to the pathogen and have a better understanding of the host defence mechanism. Two cDNA libraries, one of WSSV-challenged M. rosenbergii and a normal control one, were sequenced using the Illumina HiSeq™ 2000 platform. After de novo assembly and clustering of the unigenes from both libraries, 63,584 standard unigenes were generated with a mean size of 698bp and an N50 of 1137bp. We successfully annotated 35.31% of all unigenes by using BLASTX program (E-value <10-5) against NCBI non-redundant (Nr), Swiss-Prot, Kyoto Encyclopedia of Genes and Genome pathway (KEGG) and Orthologous Groups of proteins (COG) databases. Gene Ontology (GO) assessment was conducted using BLAST2GO software. Differentially expressed genes (DEGs) by using the FPKM method showed 8443 host genes were significantly up-regulated whereas 5973 genes were significantly down-regulated. The differentially expressed immune related genes were grouped into 15 animal immune functions. The present study showed that WSSV infection has a significant impact on the transcriptome profile of M. rosenbergii's hepatopancreas, and further enhanced the knowledge of this host-virus interaction. Furthermore, the high number of transcripts generated in this study will provide a platform for future genomic research on freshwater prawns. Copyright © 2016 Elsevier Inc. All rights reserved.

  4. Fine-scale genetic variation and evolution of West Nile Virus in a transmission "hot spot" in suburban Chicago, USA.

    Science.gov (United States)

    Bertolotti, Luigi; Kitron, Uriel D; Walker, Edward D; Ruiz, Marilyn O; Brawn, Jeffrey D; Loss, Scott R; Hamer, Gabriel L; Goldberg, Tony L

    2008-05-10

    Mosquitoes and birds were sampled for West Nile virus (WNV) in suburban Chicago, USA, in a "hot spot" of arboviral transmission. Viral genetic diversity within this area was similar to that within Illinois and the United States. Diversity was higher among viruses from mosquitoes than from birds, higher among viruses from birds in urban "green spaces" than from birds in residential areas, but lower among viruses from mosquitoes in green spaces than from mosquitoes in residential areas. Viral transmission was distance-limited, as evidenced by decreasing autocorrelation of WNV sequences with increasing geographic separation. The evolutionary rate of WNV within the study area between 21 July and 4 October 2005 was ten times higher than that for WNV across North America between 2002 and 2005. These results indicate that WNV transmission and evolutionary dynamics can vary seasonally and in response to fine-scale environmental conditions and landscape characteristics related to urbanization.

  5. Transcriptome changes associated with Tomato spotted wilt virus infection in various life stages of its thrips vector, Frankliniella fusca (Hinds).

    Science.gov (United States)

    Shrestha, Anita; Champagne, Donald E; Culbreath, Albert K; Rotenberg, Dorith; Whitfield, Anna E; Srinivasan, Rajagopalbabu

    2017-08-01

    Persistent propagative viruses maintain intricate interactions with their arthropod vectors. In this study, we investigated the transcriptome-level responses associated with a persistent propagative phytovirus infection in various life stages of its vector using an Illumina HiSeq sequencing platform. The pathosystem components included a Tospovirus, Tomato spotted wilt virus (TSWV), its insect vector, Frankliniella fusca (Hinds), and a plant host, Arachis hypogaea (L.). We assembled (de novo) reads from three developmental stage groups of virus-exposed and non-virus-exposed F. fusca into one transcriptome consisting of 72 366 contigs and identified 1161 differentially expressed (DE) contigs. The number of DE contigs was greatest in adults (female) (562) when compared with larvae (first and second instars) (395) and pupae (pre- and pupae) (204). Upregulated contigs in virus-exposed thrips had blastx annotations associated with intracellular transport and virus replication. Upregulated contigs were also assigned blastx annotations associated with immune responses, including apoptosis and phagocytosis. In virus-exposed larvae, Blast2GO analysis identified functional groups, such as multicellular development with downregulated contigs, while reproduction, embryo development and growth were identified with upregulated contigs in virus-exposed adults. This study provides insights into differences in transcriptome-level responses modulated by TSWV in various life stages of an important vector, F. fusca.

  6. Molecular Typing of Turkish Apple Chlorotic Leaf Spot Virus Isolates Based on Partial Coat Protein Gene

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    C. Ulubas Serce

    2006-08-01

    Full Text Available Apple chlorotic leaf spot virus (ACLSV isolates from various hosts and geographic locations in Turkey were molecularly characterized by RFLP, nucleotide sequence analysis and the construction of a phylogenetic tree including ACLSV isolates from GenBank. Based on nucleotide sequence alignment and the phylogenetic tree, we proposed a classification of ACLSV isolates in which isolates were divided into three major groups. The first group contained mainly Far-Eastern isolates, the second group the Hungarian (eastern-European ACLSV isolates, and the third group, which contained isolates of variable characteristics, was again divided into two subgroups, subgroup I containing mixed European isolates, and subgroup II containing central European isolates. Three representative Turkish ACLSV isolates belonged to the third group; of these, one was from the mixed European cluster (subgroup I and two from the central European cluster (subgroup II. The nucleotide sequence divergence and geographic origin of the ACLSV isolates were correlated, which indicated the possible extraction of the Turkish isolates.

  7. Pattern of the Occurrence of Tomato spotted wilt virus in Jeonnam Province

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    Sug-Ju Ko

    2013-12-01

    Full Text Available Tomato spotted wilt virus (TSWV was occurred at 8 areas including Naju, Suncheon, Younggwang, Youngam, and Shinan in Jeonnam province and the crops of Younggwang were severely damaged by TSWV. The hot pepper (Capsicum annuum, bell pepper (Capsicum annuum v ar. angulosum and tomato (Solanum lycopersicon in greenhouse and hot pepper in open field were infected by TSWV. Especially, hot pepper was severely damaged by TSWV infection. The survey data indicated that 1.1−30% in the nursery field at Naju, Suncheon, and Jangheung were infected by TSWV. Plants were infected by TSWV from early June to August. However, TSWV-infected seedlings from nursery fields showed the disease symptoms from May after transplanting. In pepper greenhouses, Frankliniella occidentalis was more dominant insect vector than Frankliniella intonsa. But in open field, the population of insect vector was opposed to greenhouse. In addition, the removal of weeds was able to delay the incidence of TSWV via side-window of greenhouse in Winter. Taken together, the control of weed and insect vector nearby side-window of greenhouse is important to prevent TSWV infection of plants.

  8. Effectiveness of combining resistance to Thielaviopsis basicola and Tomato spotted wilt virus in haploid tobacco genotypes.

    Science.gov (United States)

    Trojak-Goluch, Anna; Laskowska, Dorota; Agacka, Monika; Czarnecka, Diana; Kawka, Magdalena; Czubacka, Anna

    2011-12-01

    Black root rot (BRR) caused by Thielaviopsis basicola as well as Tomato spotted wilt virus (TSWV) are the most serious problems in tobacco growing regions. We crossed the breeding line WGL 3 carrying BRR resistance derived from N.glauca with the line PW-834 the resistance of which to TSWV was transferred from cultivar Polalta. Anthers obtained from F(1) hybrid plants were cultured to induce haploids combining resistance to Th. basicola and TSWV. Flow cytometry analysis revealed 242 haploids and 2 spontaneous doubled haploids among regenerants. All haploids were cloned and then evaluated for BRR as well as TSWV resistance. The presence of pathogens was detected by microscopic evaluation of roots or using DAS-ELISA test. Microscopic assessment showed that, 132 haploids had no symptoms of Th. basicola which, together with the absence of symptoms in the F(1) hybrids, indicated a dominant monogenic mode of inheritance. At the same time only 30 haploids demonstrated resistance to TSWV. SCAR markers associated with TSWV resistance gene detection was applied. The results indicate that small proportion of TSWV-resistant haploids is probably due to the influence of deleterious genes flanking the resistance factor that reduced vitality of gametophytes. Altogether, 24 haploids showed multiple resistance to Th. basicola and TSWV.

  9. White spot syndrome virus strains of different virulence induce distinct immune response in Cherax quadricarinatus.

    Science.gov (United States)

    Gao, Meiling; Li, Fang; Xu, Limei; Zhu, Xiaoming

    2014-07-01

    In this study, we identified three white spot syndrome virus (WSSV) strains (WSSV-CN01, WSSV-CN02 and WSSV-CN03) with significant differences in virulence. Among them, WSSV-CN01 caused significant higher and earlier mortality in redclaw crayfish Cherax quadricarinatus, thus was determined as high-virulent, while WSSV-CN02 and WSSV-CN03 were moderate-virulent and low-virulent. By investigating the total number of the circulating haemocytes and the activity of immune relative enzymes, we demonstrated that the different virulent WSSV strains induced distinct immune response in the host. Notably, a dramatic reduction of circulating haemocytes was observed in the crayfish infected with WSSV-CN01 and WSSV-CN02 but not WSSV-CN03. Further analysis revealed that cell death induced by WSSV-CN01 and WSSV-CN02 might be responsible for the decrease of circulating haemocytes. Copyright © 2014 Elsevier Ltd. All rights reserved.

  10. Metabolic product response profiles of Cherax quadricarinatus towards white spot syndrome virus infection.

    Science.gov (United States)

    Fan, Weiwei; Ye, Yangfang; Chen, Zhen; Shao, Yina; Xie, Xiaolu; Zhang, Weiwei; Liu, Hai-Peng; Li, Chenghua

    2016-08-01

    White spot syndrome virus (WSSV) is one of the most devastating viral pathogens in both shrimp and crayfish farms, which often causes disease outbreak and leads to massive moralities with significant economic losses of aquaculture. However, limited research has been carried out on the intrinsic mechanisms toward WSSV challenge at the metabolic level. To gain comprehensive insight into metabolic responses induced by WSSV, we applied an NMR approach to investigate metabolic changes of crayfish gill and hepatopancreas infected by WSSV for 1, 6 and 12 h. In gill, an enhanced energy metabolism was observed in WSSV-challenged crayfish samples at 1 h, as marked by increased glucose, alanine, methionine, glutamate and uracil. Afterwards, energy metabolism, lipid metabolism as well as osmoregulation were markedly increased at 6 hpi, as shown by elevated glucose, alanine, methionine, fumarate, tyrosine, tryptophan, histidine, phosphorylcholine, betaine and uracil, whereas no obvious metabolites change was detected at 12 hpi. As for hepatopancreas, disturbed lipid metabolism and induced osmotic regulation was found at 6 hpi based on the metabolic biomarkers such as branched chain amino acids, threonine, alanine, methionine, glutamate, glutamine, tyrosine, phenylalanine, lactate and lipid. However, no obvious metabolic change was shown in hepatopancreas at both 1 hpi and 12 hpi. Taken together, our present results provided essential metabolic information about host-pathogen interactions in crayfish, which shed new light on our understanding of WSSV infection at metabolic level. Copyright © 2016 Elsevier Ltd. All rights reserved.

  11. Increased nucleoside diphosphate kinase activity induces white spot syndrome virus infection in Litopenaeus vannamei.

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    Peng-Fei Liu

    Full Text Available Nucleoside diphosphate kinase (NDK, which has the same sequence as oncoprotein (OP in humans, can induce nucleoside triphosphates in DNA replication by maintenance of the deoxynucleotide triphosphate (dNTP's and is known to be regulated by viral infection in the shrimp Litopenaeus vannamei. This paper describes the relationship between NDK and white spot syndrome virus (WSSV infection. The recombinant NDK was produced by a prokaryotic expression system. WSSV copy numbers and mRNA levels of IE1 and VP28 were significantly increased in shrimp injected with recombinant NDK at 72 h after WSSV infection. After synthesizing dsRNA-NDK and confirming the efficacy of NDK silencing, we recorded the cumulative mortality of WSSV-infected shrimp injected with NDK and dsRNA-NDK. A comparison between the results demonstrated that silencing NDK delayed the death of shrimps. These findings indicate that NDK has an important role influencing the replication of WSSV replication in shrimp. Furthermore, NDK may have potential target as a new therapeutic strategy against WSSV infection in shrimp.

  12. Serological and molecular characterization of Syrian Tomato spotted wilt virus isolates

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    Faiz ISMAEIL

    2015-04-01

    Full Text Available Thirty four Syrian isolates of Tomato spotted wilt virus (TSWV collected from tomato and pepper were tested against five specific monoclonal antibodies using TAS-ELISA. The isolates were in two serogroups. Fourteen tomato and sixteen pepper isolates were similar in their reaction with MAb-2, MAb-4, MAb-5 and MAb-6, but did not react with MAb-7 (Serogroup 1. Meanwhile, four isolates collected from pepper reacted with all the MAbs used (Serogroup 2. The expected 620 bp DNA fragment was obtained by RT-PCR from six samples using a specific primer pair designed to amplify the nucleocapsid protein (NP gene of TSWV. The PCR products were sequenced and a phylogenetic tree was constructed. Sequence analysis revealed that the Syrian TSWV isolates were very similar at the nucleotide (97.74 to 99.84% identity and amino acid (96.17 to 99.03% identity sequences levels. The phylogenetic tree showed high similarity of Syrian TSWV isolates with many other representative isolates from different countries.

  13. Pelargonium zonate spot virus is transmitted vertically via seed and pollen in tomato.

    Science.gov (United States)

    Lapidot, M; Guenoune-Gelbart, D; Leibman, D; Holdengreber, V; Davidovitz, M; Machbash, Z; Klieman-Shoval, S; Cohen, S; Gal-On, A

    2010-08-01

    In autumn 2007, a new disease with unknown etiology was observed in open-field tomato (Solanum lycopersicum) in the Lachish region of Israel. The symptoms included mild mosaic, leaf malformation, and severe stunting of the plants. The causal agent was readily transmitted mechanically from the sap of infected plants to indicator plants. Viral particles were purified from infected plants and cDNA was synthesized from RNA isolated from the particles. Cloning and sequencing of the cDNA showed 95% identity to RNA 3 of Pelargonium zonate spot virus (PZSV). Using reverse-transcription polymerase chain reaction, PZSV was detected in both seed and pollen grains of infected tomato plants. Attempts to disinfect seed by using hydrochloric acid and trisodium phosphate failed to eliminate this PZSV detection. Seed from infected tomato plants gave rise to infected seedlings with a seed-transmission rate of PZSV of 11 to 29%. Pollen grains collected from flowers of infected plants were used to hand pollinate healthy mother tomato plants. Although none of the pollinated mother plants became infected with PZSV, 29% of the seedlings produced from seed harvested from these plants were found to be infected. This is the first demonstration that PZSV is transmitted vertically via both pollen and seed in tomato plants.

  14. Molecular markers linked to papaya ring spot virus resistance and Fusarium race 2 resistance in melon.

    Science.gov (United States)

    Brotman, Yariv; Kovalski, Irina; Dogimont, Catherine; Pitrat, Michel; Portnoy, Vitaly; Katzir, Nurit; Perl-Treves, Rafael

    2005-01-01

    In melon, the Fom-1 gene confers monogenic resistance against the soil-borne fungus Fusarium oxysporum f. sp. melonis, races 0 and 2, while the closely linked Prv gene specifies resistance against the papaya ring spot virus. Markers linked to these resistance (R) genes were identified using two recombinant inbred line populations, derived from crosses between Cucumis melo Vedrantais and C. melo PI 161375, and between C. melo Vedrantais and C. melo PI 414723, respectively. Using bulked segregant analysis, as well as systematic scoring of the mapping populations, we developed two amplified fragment length polymorphism markers, two random amplified polymorphic DNA markers and five restriction fragment length polymorphism (RFLP) markers linked to this locus. Four of the RFLP sequences bear homology to nucleotide-binding site-leucine-rich repeat R genes, indicating the presence of a significant R-gene cluster in this locus. Our study provides the most closely linked markers published so far for these important traits. It also improves the resolution of the whole linkage group IX, which was difficult to order in our previous studies. Two of the markers were converted to cleaved amplified polymorphic sequence markers to facilitate their application in marker-assisted selection. Testing these two markers in several melon lines revealed different marker haplotypes in the melon germplasm and supported multiple, independent origin of the Fusarium races 0 and 2 resistance trait.

  15. Iris Yellow Spot Virus in the Netherlands: Occurence in Onion and Confirmation of Transmission by Thrips tabaci

    OpenAIRE

    Hoedjes, K.; Verhoeven, J. Th. J.; Goldbach, R.W.; Peters, D.

    2011-01-01

    Since its first detection in the Netherlands in 1992, Iris yellow spot virus (IYSV, genus Tospovirus) has been reported worldwide in Allium crops, in a few ornamentals and in a small number of weeds. After recent findings of IYSV in Alstroemeria and Eustoma in the Netherlands, a number of neighbouring onion fields were surveyed. In 2005 and 2006, only few infected plants were found with obvious symptoms of IYSV. In 2007, after sampling and testing small leaf samples with various types of dama...

  16. Temporal Dynamics of Iris Yellow Spot Virus and Its Vector, Thrips tabaci (Thysanoptera: Thripidae), in Seeded and Transplanted Onion Fields

    OpenAIRE

    Hsu, Cynthia L.; Hoepting, Christine A.; Fuchs, Marc; Shelton, Anthony M.; Nault, Brian A.

    2017-01-01

    Onion thrips, Thrips tabaci (Lindeman) (Thysanoptera: Thripidae), can reduce onion bulb yield and transmit iris yellow spot virus (IYSV) (Bunyaviridae: Tospovirus), which can cause additional yield losses. In New York, onions are planted using seeds and imported transplants. IYSV is not seed transmitted, but infected transplants have been found in other U.S. states. Transplants are also larger than seeded onions early in the season, and thrips, some of which may be viruliferous, may preferent...

  17. Mud crab susceptibility to disease from white spot syndrome virus is species-dependent

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    Sritunyalucksana Kallaya

    2010-11-01

    Full Text Available Abstract Background Based on a report for one species (Scylla serrata, it is widely believed that mud crabs are relatively resistant to disease caused by white spot syndrome virus (WSSV. We tested this hypothesis by determining the degree of susceptibility in two species of mud crabs, Scylla olivacea and Scylla paramamosain, both of which were identified by mitochondrial 16 S ribosomal gene analysis. We compared single-dose and serial-dose WSSV challenges on S. olivacea and S. paramamosain. Findings In a preliminary test using S. olivacea alone, a dose of 1 × 106 WSSV copies/g gave 100% mortality within 7 days. In a subsequent test, 17 S. olivacea and 13 S. paramamosain were divided into test and control groups for challenge with WSSV at 5 incremental, biweekly doses starting from 1 × 104 and ending at 5 × 106 copies/g. For 11 S. olivacea challenged, 3 specimens died at doses between 1 × 105 and 5 × 105 copies/g and none died for 2 weeks after the subsequent dose (1 × 106 copies/g that was lethal within 7 days in the preliminary test. However, after the final challenge on day 56 (5 × 106 copies/g, the remaining 7 of 11 S. olivacea (63.64% died within 2 weeks. There was no mortality in the buffer-injected control crabs. For 9 S. paramamosain challenged in the same way, 5 (55.56% died after challenge doses between 1 × 104 and 5 × 105 copies/g, and none died for 2 weeks after the challenge dose of 1 × 106 copies/g. After the final challenge (5 × 106 copies/g on day 56, no S. paramamosain died during 2 weeks after the challenge, and 2 of 9 WSSV-infected S. paramamosain (22.22% remained alive together with the control crabs until the end of the test on day 106. Viral loads in these survivors were low when compared to those in the moribund crabs. Conclusions S. olivacea and S. paramamosain show wide variation in response to challenge with WSSV. S. olivacea and S. paramamosain are susceptible to white spot disease, and S. olivacea is more

  18. Mud crab susceptibility to disease from white spot syndrome virus is species-dependent.

    Science.gov (United States)

    Somboonna, Naraporn; Mangkalanan, Seksan; Udompetcharaporn, Attasit; Krittanai, Chartchai; Sritunyalucksana, Kallaya; Flegel, Tw

    2010-11-20

    Based on a report for one species (Scylla serrata), it is widely believed that mud crabs are relatively resistant to disease caused by white spot syndrome virus (WSSV). We tested this hypothesis by determining the degree of susceptibility in two species of mud crabs, Scylla olivacea and Scylla paramamosain, both of which were identified by mitochondrial 16 S ribosomal gene analysis. We compared single-dose and serial-dose WSSV challenges on S. olivacea and S. paramamosain. In a preliminary test using S. olivacea alone, a dose of 1 × 106 WSSV copies/g gave 100% mortality within 7 days. In a subsequent test, 17 S. olivacea and 13 S. paramamosain were divided into test and control groups for challenge with WSSV at 5 incremental, biweekly doses starting from 1 × 104 and ending at 5 × 106 copies/g. For 11 S. olivacea challenged, 3 specimens died at doses between 1 × 105 and 5 × 105 copies/g and none died for 2 weeks after the subsequent dose (1 × 106 copies/g) that was lethal within 7 days in the preliminary test. However, after the final challenge on day 56 (5 × 106 copies/g), the remaining 7 of 11 S. olivacea (63.64%) died within 2 weeks. There was no mortality in the buffer-injected control crabs. For 9 S. paramamosain challenged in the same way, 5 (55.56%) died after challenge doses between 1 × 104 and 5 × 105 copies/g, and none died for 2 weeks after the challenge dose of 1 × 106 copies/g. After the final challenge (5 × 106 copies/g) on day 56, no S. paramamosain died during 2 weeks after the challenge, and 2 of 9 WSSV-infected S. paramamosain (22.22%) remained alive together with the control crabs until the end of the test on day 106. Viral loads in these survivors were low when compared to those in the moribund crabs. S. olivacea and S. paramamosain show wide variation in response to challenge with WSSV. S. olivacea and S. paramamosain are susceptible to white spot disease, and S. olivacea is more susceptible than S. paramamosain. Based on our single

  19. Characterization of a New Tomato Spotted Wilt Virus Isolates Found in Hippeastrum hybridum (Hort. Plants in Poland

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    Berniak Hanna

    2016-06-01

    Full Text Available Two Tomato spotted wilt virus (TSWV isolates H1 and H2 found in Hippeastrum hybridum plants were characterized based on biological, serological, and molecular properties. Virus isolates showed differences in symptom expression – H1 isolate displayed severe necrotic spots and patterns, whereas mild mosaic symptoms were observed on H2-infected H. hybridum plants. Both TSWV isolates showed comparable reactivity with TSWV-specific antibodies and they induced similar symptoms on herbaceous indicator plants, but some differences between these isolates were detected at the nucleotide sequence level of genomic S and M ssRNAs segment fragments. The nucleotide sequences encoding nucleocapsid (N and nonstructural (NSs and NSm proteins showed 98.2%, 97.5%, and 96.5% identity, respectively. Phylogenetic analysis of N and NSs sequences conducted for tested isolates and 31 TSWV isolates included for comparison revealed that H1 and H2 isolates fell into the same cluster and they were grouped together with isolates found previously in different vegetables, ornamentals, and weeds. When NSm ORF was analyzed, the tested isolates formed a separate cluster: H1 isolate showed the highest affinity with TSWV isolates infecting chrysanthemum and pepper plants, whereas H2 isolate was most closely related to other virus isolates found in sweet pepper and tomatoes. These results indicate that both isolates were reassortants between different virus isolates, and represented two novel genetic patterns of TSWV.

  20. Evaluation of onion cultivars for resistance to onion thrips (Thysanoptera: Thripidae) and Iris yellow spot virus.

    Science.gov (United States)

    Diaz-Montano, John; Fuchs, Marc; Nault, Brian A; Shelton, Anthony M

    2010-06-01

    Onion thrips, Thrips tabaci Lindeman (Thysanoptera: Thripidae), a worldwide pest of onion, Allium cepa L., can reduce onion yield by > 50% and be even more problematic when it transmits Iris yellow spot virus (family Bunyaviridae, genus Tospovirus, IYSV). Because T. tabaci is difficult to control with insecticides and other strategies, field studies on onion, Allium cepa L., resistance to T. tabaci and IYSV were conducted in 2007 and 2008 in two locations in New York state. Forty-nine cultivars were evaluated for resistance by counting the number of larvae weekly and recording leaf damage. In another experiment, the impact of T. tabaci and IYSV on plant growth and yield was examined by spraying half of the plants with an insecticide. Eleven of the 49 cultivars had very little leaf damage and were considered resistant to T. tabaci. Visual assessment indicated that all resistant cultivars had yellow-green- colored foliage, whereas the other 38 had blue-green- colored foliage. The visual assessment of color agreed with data on color taken with a HunterLab Ultra Scan XE colorimeter. The onions 'Colorado 6' and 'NMSU 03-52-1' had the lowest numbers of T. tabaci, suggesting strong antibiosis and/or antixenosis. The other nine cultivars had variable numbers of T. tabaci, indicating a possible combination of categories of resistance. In the nonprotected treatments there were significant reductions in plant height and plant weight in most of the resistant cultivars, but there were reductions in bulb weight only in a few of them. The average of plants infected with IYSV was 10% in 2007 and 60% in 2008. Our findings indicate potential for developing onion resistance to T. tabaci as part of an overall integrated pest management strategy but suggest difficulties in identifying resistance to IYSV.

  1. Molecular immune response of the American lobster (Homarus americanus) to the White Spot Syndrome Virus.

    Science.gov (United States)

    Clark, K Fraser; Greenwood, Spencer J; Acorn, Adam R; Byrne, Philip J

    2013-11-01

    The adult American lobster (Homarus americanus) is susceptible to few naturally occurring pathogens, and no viral pathogen is known to exist. Despite this, relatively little is known about the H. americanus immune system and nothing is known about its potential viral immune response. Hundreds of rural communities in Atlantic Canada rely on the lobster fishery for their economic sustainability and could be devastated by large-scale pathogen-mediated mortality events. The White Spot Syndrome Virus is the most economically devastating viral pathogen to global shrimp aquaculture production and has been proposed to be capable of infecting all decapod crustaceans including the European Lobster. An in vivo WSSV injection challenge was conducted in H. americanus and WSSV was found to be capable of infecting and replicating within lobsters held at 20°C. The in vivo WSSV challenge also generated the first viral disease model of H. americanus and allowed for the high-throughput examination of transcriptomic changes that occur during viral infection. Microarray analysis found 136 differentially expressed genes and the expression of a subset of these genes was verified using RT-qPCR. Anti-lipopolysaccharide isoforms and acute phase serum amyloid protein A expression did not change during WSSV infection, contrary to previous findings during bacterial and parasitic infection of H. americanus. This, along with the differential gene expression of thioredoxin and trypsin isoforms, provides compelling evidence that H. americanus is capable of mounting an immune response specific to infection by different pathogen classes. Copyright © 2013 Elsevier Inc. All rights reserved.

  2. Inhibitory effect of Distamycin-A and a pyrazino-pyrazine derivative on tomato spotted wilt virus.

    Science.gov (United States)

    De Fazio, G; Kudamatsu, M

    1983-08-01

    Distamycin-A hydrochloride, a synthetic antibiotic, and 2,3-dihydroxy-6-bromo-pyrazino (2,3-beta) pyrazine derivative, were used against tomato spotted wilt virus (TSWV) in tobacco plants. The drugs were applied to the leaves at concentrations of 200 and 400 mg/l. The results showed that both drugs delayed virus spread within the plant, retarding the appearance of systemic symptoms. A virus recovery test, carried out on primary leaves of Phaseolus vulgaris cv. Manteiga, showed that TSWV replication was markedly inhibited by the pyrazino-pyrazine derivative at concentrations of 200 and 400 mg/l and, to a lower extent, by Dystamycin-A at 400 mg/l.

  3. Development of a microarray for simultaneous detection and differentiation of different tospoviruses that are serologically related to Tomato spotted wilt virus.

    Science.gov (United States)

    Liu, Lu-Yuan; Ye, He-Yi; Chen, Tsang-Hai; Chen, Tsung-Chi

    2017-01-10

    Tospoviruses, the plant-infecting genus in the family Bunyaviridae, are thrips borne and cause severe agricultural losses worldwide. Based on the serological relationships of the structural nucleocapsid protein (NP), the current tospoviruses are divided into six serogroups. The use of NP-antisera is convenient for virus detection, but it is insufficient to identify virus species grouped in a serogroup due to the serological cross-reaction. Alternatively, virus species can be identified by the N gene amplification using specific primers. Tomato spotted wilt virus (TSWV) is the type species of the genus Tospovirus and one of the most destructive plant viruses. Eight known tospoviruses, Alstroemeria necrotic streak virus (ANSV), Chrysanthemum stem necrosis virus (CSNV), Groundnut ringspot virus (GRSV), Impatiens necrotic spot virus (INSV), Melon severe mosaic virus (MeSMV), Pepper necrotic spot virus (PNSV), Tomato chlorotic spot virus (TCSV) and Zucchini lethal chlorosis virus (ZLCV), sharing serological relatedness with TSWV in NP, are grouped in the TSWV serogroup. Most of the TSWV-serogroup viruses prevail in Europe and America. An efficient diagnostic method is necessary for inspecting these tospoviruses in Asia, including Taiwan. A microarray platform was developed for simultaneous detection and identification of TSWV-serogroup tospoviruses. Total RNAs extracted from Chenopodium quinoa leaves separately inoculated with ANSV, CSNV, GRSV, INSV, TCSV and TSWV were used for testing purposes. The 5'-biotinylated degenerate forward and reverse primers were designed from the consensus sequences of N genes of TSWV-serogroup tospoviruses for reverse transcription-polymerase chain reaction (RT-PCR) amplification. Virus-specific oligonucleotide probes were spotted on the surface of polyvinyl chloride (PVC) chips to hybridize with PCR products. The hybridization signals were visualized by hydrolysis of NBT/BCIP with streptavidine-conjugated alkaline phosphatase. The

  4. Specific Insect-Virus Interactions Are Responsible for Variation in Competency of Different Thrips tabaci Isolines to Transmit Different Tomato Spotted Wilt Virus Isolates

    Science.gov (United States)

    Jacobson, Alana L.; Kennedy, George G.

    2013-01-01

    Local adaptation between sympatric host and parasite populations driven by vector genetics appears to be a factor that influences dynamics of disease epidemics and evolution of insect-vectored viruses. Although T. tabaci is the primary vector of Tomato spotted wilt virus (TSWV) in some areas of the world, it is not an important vector of this economically important plant virus in many areas where it occurs. Previous studies suggest that genetic variation of thrips populations, virus isolates, or both are important factors underlying the localized importance of this species as a vector of TSWV. This study was undertaken to quantify variation in transmissibility of TSWV isolates by T. tabaci, in the ability of T. tabaci to transmit isolates of TSWV, and to examine the possibility that genetic interactions and local adaptation contribute to the localized nature of this species as a vector of TSWV. Isofemale lines of Thrips tabaci from multiple locations were tested for their ability to transmit multiple TSWV isolates collected at the same and different locations as the thrips. Results revealed that the probability of an isofemale line transmitting TSWV varied among virus isolates, and the probability of an isolate being transmitted varied among isofemale lines. These results indicate that the interaction of T. tabaci and TSWV isolate genetic determinants underlie successful transmission of TSWV by T. tabaci. Further analysis revealed sympatric vector-virus pairing resulted in higher transmission than allopatric pairing, which suggests that local adaptation is occurring between T. tabaci and TSWV isolates. PMID:23358707

  5. The NSs protein of tomato spotted wilt virus is required for persistent infection and transmission by Frankliniella occidentalis.

    Science.gov (United States)

    Margaria, P; Bosco, L; Vallino, M; Ciuffo, M; Mautino, G C; Tavella, L; Turina, M

    2014-05-01

    Tomato spotted wilt virus (TSWV) is the type member of tospoviruses (genus Tospovirus), plant-infecting viruses that cause severe damage to ornamental and vegetable crops. Tospoviruses are transmitted by thrips in the circulative propagative mode. We generated a collection of NSs-defective TSWV isolates and showed that TSWV coding for truncated NSs protein could not be transmitted by Frankliniella occidentalis. Quantitative reverse transcription (RT)-PCR and immunostaining of individual insects detected the mutant virus in second-instar larvae and adult insects, demonstrating that insects could acquire and accumulate the NSs-defective virus. Nevertheless, adults carried a significantly lower viral load, resulting in the absence of transmission. Genome sequencing and analyses of reassortant isolates showed genetic evidence of the association between the loss of competence in transmission and the mutation in the NSs coding sequence. Our findings offer new insight into the TSWV-thrips interaction and Tospovirus pathogenesis and highlight, for the first time in the Bunyaviridae family, a major role for the S segment, and specifically for the NSs protein, in virulence and efficient infection in insect vector individuals. Our work is the first to show a role for the NSs protein in virus accumulation in the insect vector in the Bunyaviridae family: demonstration was obtained for the system TSWV-F. occidentalis, arguably one of the most damaging combination for vegetable crops. Genetic evidence of the involvement of the NSs protein in vector transmission was provided with multiple approaches.

  6. Susceptibility of Chinese Perch Brain (CPB Cell and Mandarin Fish to Red-Spotted Grouper Nervous Necrosis Virus (RGNNV Infection

    Directory of Open Access Journals (Sweden)

    Jiagang Tu

    2016-05-01

    Full Text Available Nervous necrosis virus (NNV is the causative agent of viral encephalopathy and retinopathy (VER, a neurological disease responsible for high mortality of fish species worldwide. Taking advantage of our established Chinese perch brain (CPB cell line derived from brain tissues of Mandarin fish (Siniperca chuatsi, the susceptibility of CPB cell to Red-Spotted Grouper nervous necrosis virus (RGNNV was evaluated. The results showed that RGNNV replicated well in CPB cells, resulting in cellular apoptosis. Moreover, the susceptibility of Mandarin fish to RGNNV was also evaluated. Abnormal swimming was observed in RGNNV-infected Mandarin fish. In addition, the cellular vacuolation and viral particles were also observed in brain tissues of RGNNV-infected Mandarin fish by Hematoxylin-eosin staining or electronic microscopy. The established RGNNV susceptible brain cell line from freshwater fish will pave a new way for the study of the pathogenicity and replication of NNV in the future.

  7. The evidence of bacilliform virus a causative agent of white spot syndrome of white shrimp Penaeus merguiensis

    Directory of Open Access Journals (Sweden)

    Hari Suprapto

    2003-06-01

    Full Text Available The rod-shape virus particles were found in the hyperthrophied nucleus and cytoplasm of diseased white shrimpPenaeus merguiensis naturally infected by White spot syndrome (WSBV. In natural infection cumulative mortality of shrimp werevery high, 95% of population were dead in 3-7 days, and 5% of shrimp population survived. The disease was outbreak in intensiveculture farms used the closed circulation sea water system. Others crustacean such as crabs and bentatos were not infected by WSBVat time of sampling. The virus particle were rod-shape ranging from 78 ± 10 nm in diameter and 280 ± 10 nm in length. The pathogenicbacteria mainly dominated by genus Vibrio sp were isolated from shrimp.

  8. Characterization of Citrus tristeza virus isolates from grapefruit (Citrus paradisi Macf. accessions of Citrus Active Germplasm Bank Caracterização de isolados do vírus da tristeza dos citros de acessos de pomelos (Citrus paradisi Macf. do Banco Ativo de Germoplasma de Citros

    Directory of Open Access Journals (Sweden)

    Maria Júlia Corazza-Nunes

    2006-09-01

    Full Text Available Citrus tristeza virus (CTV isolates from 35 grapefruit accessions belonging to Citrus Active Germplasm Bank of the "Instituto Agronômico de Campinas" located at the "Centro APTA Citros Sylvio Moreira", Cordeirópolis, São Paulo state, Brazil, were characterized and evaluated through symptoms in the trees, biological indexing, immunological diagnosis with different monoclonal antibodies and SSCP analysis (single-strand conformation polymorphism of the coat protein gene. Symptomatology indicated that, in general, the group of plants with smaller canopy volume and severe stem pitting differed significantly from the group that presented greater vegetative development and mild to moderate stem pitting. However, the isolates from most of the accessions induced mild reaction on Mexican lime. The serological evaluation through the DAS-ELISA using monoclonal antibodies did not reveal any association between virus titer in the plant tissue and symptoms. The reaction with different monoclonal antibodies and the distinct electrophoresis patterns obtained through SSCP showed that there is a high degree of diversity among the isolates that infect these grapefruit accessions. High complexity within the same isolate was also observed in the SSCP profiles. This finding indicates that the CTV isolates from these plants are a complex mixture of CTV haplotypes. Similar SSCP banding patterns were observed among some plants with strong stem pitting symptoms, and among some plants with weak or moderate stem pitting symptoms.Isolados do vírus da tristeza dos citros (CTV de 35 acessos de pomelos que fazem parte do Banco Ativo de Germoplasma de Citros, localizado no Centro APTA Citros Sylvio Moreira, Cordeirópolis, São Paulo, Brasil, pertencente ao Instituto Agronômico de Campinas (IAC, foram caracterizados através dos sintomas observados nas árvores, indexação biológica, diagnóstico imunológico e análise SSCP (polimorfismo de conformação de fita simples

  9. Differential expression of tomato spotted wilt virus-derived viral small RNAs in infected commercial and experimental host plants.

    Directory of Open Access Journals (Sweden)

    Neena Mitter

    Full Text Available BACKGROUND: Viral small RNAs (vsiRNAs in the infected host can be generated from viral double-stranded RNA replicative intermediates, self-complementary regions of the viral genome or from the action of host RNA-dependent RNA polymerases on viral templates. The vsiRNA abundance and profile as well as the endogenous small RNA population can vary between different hosts infected by the same virus influencing viral pathogenicity and host response. There are no reports on the analysis of vsiRNAs of Tomato spotted wilt virus (TSWV, a segmented negative stranded RNA virus in the family Bunyaviridae, with two of its gene segments showing ambisense gene arrangement. The virus causes significant economic losses to numerous field and horticultural crops worldwide. PRINCIPAL FINDINGS: Tomato spotted wilt virus (TSWV-specific vsiRNAs were characterized by deep sequencing in virus-infected experimental host Nicotiana benthamiana and a commercial, susceptible host tomato. The total small (s RNA reads in TSWV-infected tomato sample showed relatively equal distribution of 21, 22 and 24 nt, whereas N. benthamiana sample was dominated by 24 nt total sRNAs. The number of vsiRNA reads detected in tomato was many a magnitude (~350:1 higher than those found in N. benthamiana, however the profile of vsiRNAs in terms of relative abundance 21, 22 and 24 nt class size was similar in both the hosts. Maximum vsiRNA reads were obtained for the M RNA segment of TSWV while the largest L RNA segment had the least number of vsiRNAs in both tomato and N. benthamiana. Only the silencing suppressor, NSs, of TSWV recorded higher antisense vsiRNA with respect to the coding frame among all the genes of TSWV. SIGNIFICANCE: Details of the origin, distribution and abundance of TSWV vsiRNAs could be useful in designing efficient targets for exploiting RNA interference for virus resistance. It also has major implications toward our understanding of the differential processing of vsi

  10. Differential expression of tomato spotted wilt virus-derived viral small RNAs in infected commercial and experimental host plants.

    Science.gov (United States)

    Mitter, Neena; Koundal, Vikas; Williams, Sarah; Pappu, Hanu

    2013-01-01

    Viral small RNAs (vsiRNAs) in the infected host can be generated from viral double-stranded RNA replicative intermediates, self-complementary regions of the viral genome or from the action of host RNA-dependent RNA polymerases on viral templates. The vsiRNA abundance and profile as well as the endogenous small RNA population can vary between different hosts infected by the same virus influencing viral pathogenicity and host response. There are no reports on the analysis of vsiRNAs of Tomato spotted wilt virus (TSWV), a segmented negative stranded RNA virus in the family Bunyaviridae, with two of its gene segments showing ambisense gene arrangement. The virus causes significant economic losses to numerous field and horticultural crops worldwide. Tomato spotted wilt virus (TSWV)-specific vsiRNAs were characterized by deep sequencing in virus-infected experimental host Nicotiana benthamiana and a commercial, susceptible host tomato. The total small (s) RNA reads in TSWV-infected tomato sample showed relatively equal distribution of 21, 22 and 24 nt, whereas N. benthamiana sample was dominated by 24 nt total sRNAs. The number of vsiRNA reads detected in tomato was many a magnitude (~350:1) higher than those found in N. benthamiana, however the profile of vsiRNAs in terms of relative abundance 21, 22 and 24 nt class size was similar in both the hosts. Maximum vsiRNA reads were obtained for the M RNA segment of TSWV while the largest L RNA segment had the least number of vsiRNAs in both tomato and N. benthamiana. Only the silencing suppressor, NSs, of TSWV recorded higher antisense vsiRNA with respect to the coding frame among all the genes of TSWV. Details of the origin, distribution and abundance of TSWV vsiRNAs could be useful in designing efficient targets for exploiting RNA interference for virus resistance. It also has major implications toward our understanding of the differential processing of vsiRNAs in antiviral defense and viral pathogenicity.

  11. Preliminary study on haemocyte response to white spot syndrome virus infection in black tiger shrimp Penaeus monodon.

    Science.gov (United States)

    van de Braak, C B T; Botterblom, M H A; Huisman, E A; Rombout, J H W M; van der Knaap, W P W

    2002-08-29

    White spot syndrome virus (WSSV) has been a major cause of shrimp mortality in aquaculture in the past decade. In contrast to extensive studies on the morphology and genome structure of the virus, little work has been done on the defence reaction of the host after WSSV infection. Therefore, we examined the haemocyte response to experimental WSSV infection in the black tiger shrimp Penaeus monodon. Haemolymph sampling and histology showed a significant decline in free, circulating haemocytes after WSSV infection. A combination of in situ hybridisation with a specific DNA probe for WSSV and immuno-histochemistry with a specific antibody against haemocyte granules in tissue sections indicated that haemocytes left the circulation and migrated to tissues where many virus-infected cells were present. However, no subsequent haemocyte response to the virus-infected cells was detected. The number of granular cells decreased in the haematopoietic tissue of infected shrimp. In addition, a fibrous-like immuno-reactive layer appears in the outer stromal matrix of tubule walls in the lymphoid organ of infected shrimp. The role of haemocytes in shrimp defence after viral infection is discussed.

  12. A 3D model of the membrane protein complex formed by the white spot syndrome virus structural proteins.

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    Yun-Shiang Chang

    Full Text Available BACKGROUND: Outbreaks of white spot disease have had a large negative economic impact on cultured shrimp worldwide. However, the pathogenesis of the causative virus, WSSV (whit spot syndrome virus, is not yet well understood. WSSV is a large enveloped virus. The WSSV virion has three structural layers surrounding its core DNA: an outer envelope, a tegument and a nucleocapsid. In this study, we investigated the protein-protein interactions of the major WSSV structural proteins, including several envelope and tegument proteins that are known to be involved in the infection process. PRINCIPAL FINDINGS: In the present report, we used coimmunoprecipitation and yeast two-hybrid assays to elucidate and/or confirm all the interactions that occur among the WSSV structural (envelope and tegument proteins VP51A, VP19, VP24, VP26 and VP28. We found that VP51A interacted directly not only with VP26 but also with VP19 and VP24. VP51A, VP19 and VP24 were also shown to have an affinity for self-interaction. Chemical cross-linking assays showed that these three self-interacting proteins could occur as dimers. CONCLUSIONS: From our present results in conjunction with other previously established interactions we construct a 3D model in which VP24 acts as a core protein that directly associates with VP26, VP28, VP38A, VP51A and WSV010 to form a membrane-associated protein complex. VP19 and VP37 are attached to this complex via association with VP51A and VP28, respectively. Through the VP26-VP51C interaction this envelope complex is anchored to the nucleocapsid, which is made of layers of rings formed by VP664. A 3D model of the nucleocapsid and the surrounding outer membrane is presented.

  13. Tomato spotted wilt virus particle assembly : studying the role of the structural proteins in vivo

    NARCIS (Netherlands)

    Snippe, M.

    2006-01-01

    Members of the Bunyaviridae have spherical, enveloped virus particles that acquire their lipid membrane at the Golgi complex. For the animal-infecting bunyaviruses, virus assembly involves budding of ribonucleoprotein particles (RNPs) into vacuolised lumen of the Golgi complex, after which the

  14. Antiviral property of marine actinomycetes against white spot syndrome virus in penaeid shrimps

    Digital Repository Service at National Institute of Oceanography (India)

    Kumar, S.S.; Philip, R.; Achuthankutty, C.T.

    and are continually being screened for new compounds. In this communication, the results of a study made to determine the effectiveness of marine actinomycetes against the white spot disease in penaeid shrimps are presented. Twenty-five isolates of actinomycetes were...

  15. Spinach: A new natural host of Impatiens necrotic spot virus in California.

    Science.gov (United States)

    Impatiens necrotic spot tospovirus (INSV; family Bunyaviridae) was detected in a spinach (Spinacia oleracea) experimental field in Monterey County, CA in October of 2008. Spinach plants exhibiting severe stunting and with leaves that showed interveinal yellowing, thickening, and deformation were obs...

  16. The plant virus Tomato Spotted Wilt Tospovirus activates the immune system of its main insect vector, Frankliniella occidentalis.

    Science.gov (United States)

    Medeiros, Ricardo B; Resende, Renato de O; de Avila, Antonio Carlos

    2004-05-01

    Tospoviruses have the ability to infect plants and their insect vectors. Tomato spotted wilt virus (TSWV), the type species in the Tospovirus genus, infects its most important insect vector, Frankliniella occidentalis, the western flower thrips (WFT). However, no detrimental effects on the life cycle or cytopathological changes have been reported in the WFT after TSWV infection, and relatively few viral particles can be observed even several days after infection. We hypothesized that TSWV infection triggers an immune response in the WFT. Using subtractive cDNA libraries to probe WFT DNA macroarrays, we found that the WFT's immune system is activated by TSWV infection. The activated genes included (i) those encoding antimicrobial peptides, such as defensin and cecropin; (ii) genes involved in pathogen recognition, such as those encoding lectins; (iii) those encoding receptors that activate the innate immune response, such as Toll-3; and (iv) those encoding members of signal transduction pathways activated by Toll-like receptors, such as JNK kinase. Transcriptional upregulation of these genes after TSWV infection was confirmed by Northern analysis, and the kinetics of the immune response was measured over time. Several of the detected genes were activated at the same time that viral replication was first detected by reverse transcription-PCR. To our knowledge, this is the first report of the activation of an insect vector immune response by a plant virus. The results may lead to a better understanding of insects' immune responses against viruses and may help in the future development of novel control strategies against plant viruses, as well as human and animal viruses transmitted by insect vectors.

  17. Assessing Virulence and Transmission Rates of White Spot Syndrome Virus (WSSV) in Two Ecologically Important Palaemonid Shrimp

    Science.gov (United States)

    Bernard, C.; Keesee, B.; Philippoff, C.; Curran, S.; Lotz, J.; Powell, E.

    2016-02-01

    Investigators, including three REU interns, conducted an experiment to quantify parameters for an epidemiological model designed to estimate disease transmission in marine invertebrates. White spot syndrome virus (WSSV) is a highly pathogenic disease affecting commercially important penaeid shrimp fisheries worldwide. The virus devastates penaeid shrimp but other varieties of decapods may serve as reservoirs for disease by being less susceptible to WSSV or refractory to disease. Non-penaeid crustaceans are less susceptible to WSSV, and different species have variable resistance to the disease leading to different potential to serve as reservoirs for transmission of the disease to coastal penaeid fisheries. This study investigates virulence and transmission rates of WSSV in two palaemonid shrimp which are keystone members of coastal food webs, and effects of species interactions on transmission rates of WSSV are estimated in a laboratory setting as a proxy for natural habitats. Two species of grass shrimp were exposed to a Chinese strain of WSSV through feeding the test individuals with previously prepared, inoculated penaeid shrimp. Replicated tanks containing 30 animals were exposed to the virus in arenas containing one or both species for 24 hours, then isolated in 1 liter tanks and monitored. During the isolation period moribund individuals were preserved for later analysis. After 7 days all test individuals were analyzed using qPCR to determine WSSV presence and load in DNA. From these data transmission rates, mortality, and viral concentration were quantified and used as parameters in a simple epidemiological model.

  18. Establishment, characterization, and virus susceptibility of a new marine cell line from red spotted grouper (Epinephelus akaara).

    Science.gov (United States)

    Zhou, Guang-Zhou; Li, Zheng-Qiu; Yuan, Xiu-Ping; Zhang, Qi-Ya

    2007-01-01

    A marine fish cell line from the snout of red spotted grouper Epinephelus akaara, a protogynous hermaphrodite, was established, characterized, and subcultured with more than 60 passages. The grouper snout cell line (GSC) cells multiplied well in Dulbecco's modified Eagle's medium (DMEM) medium supplemented with 10% fetal bovine serum. The optimal growth temperature was 25 degrees C, and morphologically the cells were fibroblastic. Chromosome analysis revealed that the GSC cell line has a normal diploid karyotype with 2n = 8st + 40t. A virus titration study indicated that the cells were susceptible to turbot Scophthalmus Maximus rhabdovirus (SMRV) (10(8.5) TCID(50) ml(-1)), while the viral titer of frog Rana grylio virus 9807 (RGV(9807)) reached 10(3.5) TCID(50) ml(-1). The infection was confirmed by cytopathic effect (CPE), immunofluorescence, and electron microscopy experiments, which detected the viral particles in the cytoplasm of virus-infected cells, respectively. Further, significant fluorescent signals were observed when the GSC cells were transfected with pEGFP vector DNA, indicating their potential utility for transgenic and genetic manipulation studies.

  19. Production of polyclonal antibodies against Pelargonium zonate spot virus coat protein expressed in Escherichia coli and application for immunodiagnosis.

    Science.gov (United States)

    Gulati-Sakhuja, Anju; Sears, John L; Nuñez, Alberto; Liu, Hsing-Yeh

    2009-09-01

    Pelargonium zonate spot virus (PZSV) is identified recently in tomato plants in the United States. To develop serological diagnostic tools for the detection of this virus, the production of good quality antibodies is a necessity. The coat protein (CP) gene of a California isolate of PZSV was cloned into a bacterial expression vector (pTriEX-4 Ek/LIC). The plasmid pTriEX-4-PZSV-CP was transformed into Escherichia coli Rosetta 2(DE3)pLacI and the recombinant PZSV-CP was expressed as a fusion protein containing N-terminal hexa-histidine and S tags. Expressed PZSV-CP was purified under denaturing conditions by affinity chromatography yielding 3mg refolded protein per 200mL of bacterial culture, and used as an antigen for raising PZSV-CP antiserum in rabbits. Specificity of the antiserum to PZSV was shown by Western blot and ELISA. When used in Western blot analysis, the antiserum was able to detect the recombinant protein, the PZSV coat protein and PZSV infected plant samples. The antiserum was successfully used in indirect-ELISA at dilutions of up to 1:16,000 to detect PZSV in infected leaf samples. Direct ELISA was successful only with denatured antigens. This is the first report on production of polyclonal antiserum against recombinant coat protein of PZSV and its use for detection and diagnosis of virus using serological methods.

  20. CLUSTER MODEL FOR EXTENSIVE GIANT TIGER SHRIMP (Penaeus monodon Fab. TO PREVENT TRANSMISSION OF WHITE SPOT SYNDROME VIRUS

    Directory of Open Access Journals (Sweden)

    Arief Taslihan

    2015-06-01

    Full Text Available White spot syndrome virus (WSSV has become epidemic in Indonesia and affecting shrimp aquaculture interm of its production. White spot syndrome virus is transmitted from one to other ponds, through crustacean, included planktonic copepode as carrier for WSSV and through water from affected shrimp pond. A cluster model, consist of shrimp grow out ponds surrounded by non-shrimp pond as a role of biosecurity has been developed. The model aimed to prevent white spot virus transmission in extensive giant tiger shrimp pond. The study was conducted in two sites at Demak District, Central Java Province. As the treatment, a cluster consist of three shrimp ponds in site I, and two shrimp ponds in site II, each was surrounded by buffer ponds rearing only finfish. As the control, five extensive shrimp grow out ponds in site I and three shrimp grow out ponds in site II, with shrimp pond has neither applied biosecurity nor surrounded by non-shrimp pond as biosecurity as well considered as control ponds. The results found that treatment of cluster shrimp ponds surrounded by non-shrimp ponds could hold shrimp at duration of culture in the grow out pond (DOC 105.6±4.5 days significantly much longer than that of control that harvested at 60.9±16.0 days due to WSSV outbreak. Survival rate in trial ponds was 77.6±3.6%, significantly higher than that of control at 22.6±15.8%. Shrimp production in treatment ponds has total production of 425.1±146.6 kg/ha significantly higher than that of control that could only produced 54.5±47.6 kg/ha. Implementation of Better Management Practices (BMP by arranging shrimp ponds in cluster and surrounding by non-shrimp ponds proven effectively prevent WSSV transmission from traditional shrimp ponds in surrounding area.

  1. Influence of Tomato Spotted Wilt Virus Uneven Distribution on Its Serological Detection in Tomato, Pepper and Ornamentals

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    Ivana Đekić

    2008-01-01

    Full Text Available Reliable detection of plant pathogens does not only mean the development of sufficiently sensitive laboratory techniques for their routine testing. Regardless of the sensitivity of applied methods, the proper selection of samples to be tested has crucial influence on method reliability. Due to uneven distribution of Tomato spotted wilt virus (TSWV in naturally infected systemic host plants, the collection and sampling of material for assaying is acritical moment upon which the reliability of laboratory procedure depends. The effect of irregular virus distribution on its serological detection was examined in tomato, pepper and four ornamental species, as its most important host plants in our country.The reliability of virus detection, depending on its uneven distribution, was assessed by serological testing of tomato and pepper symptomatic leaves and fruits, and symptomatic and asymptomatic young and old leaves, as well as flower petals of ornamentals. Although TSWV was detected using ELISA in the majority of plants included in the experiment, the tests indicated an uneven distribution and unequal concentrations of TSWV in different parts of the plants. The virus could not be detected in a certain number of subsamples, prepared from infected tomato and pepper fruits and older ornamental leaves. The virus also could not be detected in some ornamentals and tomato plants with intensive symptoms. Conversely, the virus was detected in three ornamental plants without any symptoms. Examining the virus distribution in different plant parts indicated that the reliability of ELISA could be reached not only by sampling younger ornamental leaves, but also by preparing compound samples with as much leaves as possible, or by testing a greater number of subsamples of the tested plant. Considering a small possibility of TSWV detection in tomato and pepper fruits, the infection of these should be established by testing their leaves. Besides, the data show that

  2. Antigenic and genetic characterization of the first rabies virus isolated from the bat Eumops perotis in Brazil Caracterização antigênica e genética do primeiro isolado de vírus da raiva em morcego Eumops perotis no Brasil

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    Juliana Galera Castilho

    2008-04-01

    Full Text Available Although the main transmitters of rabies in Brazil are dogs and vampire bats, the role of other species such as insectivorous and frugivorous bats deserves special attention, as the rabies virus has been isolated from 36 bat species. This study describes the first isolation of the rabies virus from the insectivorous bat Eumops perotis. The infected animal was found in the city of Ribeirão Preto, São Paulo. The virus was identified by immunofluorescence antibody test (FAT in central nervous system (CNS samples, and the isolation was carried out in N2A cell culture and adult mice. The sample was submitted to antigenic typing using a panel of monoclonal antibodies (CDC/Atlanta/USA. The DNA sequence of the nucleoprotein gene located between nucleotides 102 and 1385 was aligned with homologous sequences from GenBank using the CLUSTAL/W method, and the alignment was used to build a neighbor-joining distance-based phylogenetic tree with the K-2-P model. CNS was negative by FAT, and only one mouse died after inoculation with a suspension from the bat's CNS. Antigenic typing gave a result that was not compatible with the patterns defined by the panel. Phylogenetic analysis showed that the virus isolated segregated into the same cluster related to other viruses isolated from insectivorous bats belonging to genus Nyctinomops ssp. (98.8% nucleotide identity with each other.No Brasil, embora os principais transmissores da raiva sejam cães e morcegos hematófagos, o papel de outras espécies, tais como morcegos insetívoros e frugívoros, merece atenção especial, uma vez que o vírus da raiva já foi isolado em 36 espécies de morcegos. Este estudo descreve o primeiro isolamento do vírus da raiva em um morcego insetívoro Eumops perotis. O animal infectado foi encontrado na cidade de Ribeirão Preto, São Paulo. O vírus foi identificado pelo teste de imunofluorescência direta (IFD em amostras de sistema nervoso central (SNC, e o isolamento foi

  3. Effects of Thrips Density, Mode of Inoculation, and Plant Age on Tomato Spotted Wilt Virus Transmission in Peanut Plants.

    Science.gov (United States)

    Shrestha, Anita; Sundaraj, Sivamani; Culbreath, Albert K; Riley, David G; Abney, Mark R; Srinivasan, Rajagopalbabu

    2015-02-01

    Spotted wilt caused by tomato spotted wilt virus (TSWV; family Bunyaviridae; genus Tospovirus) is a serious disease of peanut (Arachis hypogaea L.) in the southeastern United States. Peanut genotypes with field resistance to TSWV are effective in suppressing spotted wilt. All commercially available genotypes with field resistance to TSWV were developed through conventional breeding. As a part of the breeding process, peanut genotypes are regularly screened under field situations. Despite numerous advantages associated with field screening, it is often limited by inconsistent vector (thrips) and TSWV pressure. A greenhouse transmission protocol would aid in thorough screening of selected genotypes and conserve time. In this study, various parameters associated with TSWV transmission, including tobacco thrips, Frankliniella fusca (Hinds) density, mode of inoculation, and plant age, were evaluated. Greater incidences of TSWV infection were obtained with thrips-mediated inoculation when compared with mechanical inoculation. TSWV inoculation with three, five, and 10 thrips resulted in greater incidences of TSWV infection in plants than inoculation with one thrips. However, incidences of TSWV infection did not vary between plants inoculated with three, five, and 10 viruliferous thrips. With both thrips-mediated and mechanical inoculation methods, incidences of TSWV infection in 1-wk-old plants were greater than in 4-wk-old plants. TSWV copy numbers, as determined by qPCR, also decreased with plant age. Results suggest that using at least three thrips per plant and 1- to 2-wk-old plants would maximize TSWV infection in inoculated plants. © The Author 2015. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

  4. Indel-II region deletion sizes in the white spot syndrome virus genome correlate with shrimp disease outbreaks in southern Vietnam

    NARCIS (Netherlands)

    Tran Thi Tuyet, H.; Zwart, M.P.; Phuong, N.T.; Oanh, D.T.H.; Jong, de M.C.M.; Vlak, J.M.

    2012-01-01

    Sequence comparisons of the genomes of white spot syndrome virus (WSSV) strains have identified regions containing variable-length insertions/deletions (i.e. indels). Indel-I and Indel-II, positioned between open reading frames (ORFs) 14/15 and 23/24, respectively, are the largest and the most

  5. Thrips resistance in pepper and its consequences for the acquisition and inoculation of Tomato spotted wilt virus by the Western Flower Thrips

    NARCIS (Netherlands)

    Maris, P.C.; Joosten, N.N.; Peters, D.; Goldbach, R.W.

    2003-01-01

    Different levels of thrips resistance were found in seven Capsicum accessions. Based on the level of feeding damage, host preference, and host suitability for reproduction, a thrips susceptible and a resistant accession were selected to study their performance as Tomato spotted wilt virus (TSWV)

  6. Application of phage display in selecting Tomato spotted wilt virus - specific single-chain antibodies (scFvs) for sensitive diagnosis in ELISA

    NARCIS (Netherlands)

    Griep, R.A.; Prins, M.; Twisk, van C.; Keller, H.J.H.G.; Kerschbaumer, R.J.; Kormelink, R.; Goldbach, R.W.; Schots, A.

    2000-01-01

    A panel of recombinant single-chain antibodies (scFvs) against structural proteins of Tomato spotted wilt virus (TSWV) was retrieved from a human combinatorial scFv antibody library using the novel phage display technique. After subcloning the encoding DNA sequences in the expression vector pSKAP/S,

  7. Detection of hepatitis B virus DNA in sera from 18 alcoholic carriers of "anti-HBc alone" and response to a single dose of hepatitis B vaccine Pesquisa de DNA do vírus da hepatite B no soro de 18 alcoolistas portadores de "anti-HBc isolado" e resposta a uma dose da vacina para hepatite B

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    Luiz Carlos Marques de Oliveira

    2008-09-01

    Full Text Available To evaluate the possibility of occult hepatitis B virus (HBV infection in alcoholics carriers of "anti-HBc alone", and to verify the behavior of this serological pattern after a single dose of hepatitis B vaccine, 18 alcoholics who had this serological profile were evaluated by the polymerase chain reaction method, and 17 of them were vaccined. All were negative for HBV DNA. Nine (52.9% of those vaccined had anamnestic response, mainly those with positive anti-HBe (8/10; 80%. "Anti-HBc alone" was compatible with low levels of anti-HBs in half of the patients, and probably with false positive results for anti-HBc in the others.Para avaliar a possibilidade de infecção oculta pelo vírus da hepatite B em alcoolistas com "anti-HBc isolado" e a resposta a uma dose da vacina para a hepatite B, 18 alcoolistas com este perfil sorológico foram avaliados pelo método de reação em cadeia da polimerase e 17 deles foram vacinados. Todos tiveram negativos os exames para o VHB DNA. Nove (52,9% dos vacinados tiveram resposta anamnéstica, principalmente aqueles com anti-HBe positivo (8/10; 80%. "Anti-HBc isolado" foi compatível com baixos títulos de anti-HBs em metade dos pacientes, e provavelmente com resultado falso-positivo para o anti-HBc nos demais.

  8. Tomato spotted wilt virus benefits a non-vector arthropod, Tetranychus urticae, by modulating different plant responses in tomato.

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    Punya Nachappa

    Full Text Available The interaction between plant viruses and non-vector arthropod herbivores is poorly understood. However, there is accumulating evidence that plant viruses can impact fitness of non-vector herbivores. In this study, we used oligonucleotide microarrays, phytohormone, and total free amino acid analyses to characterize the molecular mechanisms underlying the interaction between Tomato spotted wilt virus (TSWV and a non-vector arthropod, twospotted spider mite (Tetranychusurticae, on tomato plants, Solanumlycopersicum. Twospotted spider mites showed increased preference for and fecundity on TSWV-infected plants compared to mock-inoculated plants. Transcriptome profiles of TSWV-infected plants indicated significant up-regulation of salicylic acid (SA-related genes, but no apparent down-regulation of jasmonic acid (JA-related genes which could potentially confer induced resistance against TSM. This suggests that there was no antagonistic crosstalk between the signaling pathways to influence the interaction between TSWV and spider mites. In fact, SA- and JA-related genes were up-regulated when plants were challenged with both TSWV and the herbivore. TSWV infection resulted in down-regulation of cell wall-related genes and photosynthesis-associated genes, which may contribute to host plant susceptibility. There was a three-fold increase in total free amino acid content in virus-infected plants compared to mock-inoculated plants. Total free amino acid content is critical for arthropod nutrition and may, in part, explain the apparent positive indirect effect of TSWV on spider mites. Taken together, these data suggest that the mechanism(s of increased host suitability of TSWV-infected plants to non-vector herbivores is complex and likely involves several plant biochemical processes.

  9. Sequencing and de novo analysis of the hemocytes transcriptome in Litopenaeus vannamei response to white spot syndrome virus infection.

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    Shuxia Xue

    Full Text Available BACKGROUND: White spot syndrome virus (WSSV is a causative pathogen found in most shrimp farming areas of the world and causes large economic losses to the shrimp aquaculture. The mechanism underlying the molecular pathogenesis of the highly virulent WSSV remains unknown. To better understand the virus-host interactions at the molecular level, the transcriptome profiles in hemocytes of unchallenged and WSSV-challenged shrimp (Litopenaeus vannamei were compared using a short-read deep sequencing method (Illumina. RESULTS: RNA-seq analysis generated more than 25.81 million clean pair end (PE reads, which were assembled into 52,073 unigenes (mean size = 520 bp. Based on sequence similarity searches, 23,568 (45.3% genes were identified, among which 6,562 and 7,822 unigenes were assigned to gene ontology (GO categories and clusters of orthologous groups (COG, respectively. Searches in the Kyoto Encyclopedia of Genes and Genomes Pathway database (KEGG mapped 14,941 (63.4% unigenes to 240 KEGG pathways. Among all the annotated unigenes, 1,179 were associated with immune-related genes. Digital gene expression (DGE analysis revealed that the host transcriptome profile was slightly changed in the early infection (5 hours post injection of the virus, while large transcriptional differences were identified in the late infection (48 hpi of WSSV. The differentially expressed genes mainly involved in pattern recognition genes and some immune response factors. The results indicated that antiviral immune mechanisms were probably involved in the recognition of pathogen-associated molecular patterns. CONCLUSIONS: This study provided a global survey of host gene activities against virus infection in a non-model organism, pacific white shrimp. Results can contribute to the in-depth study of candidate genes in white shrimp, and help to improve the current understanding of host-pathogen interactions.

  10. Horizontal transmission dynamics of White spot syndrome virus by cohabitation trials in juvenile Penaeus monodon and P. vannamei.

    Science.gov (United States)

    Tuyen, N X; Verreth, J; Vlak, J M; de Jong, M C M

    2014-11-01

    White spot syndrome virus (WSSV), a rod-shaped double-stranded DNA virus, is an infectious agent causing fatal disease in shrimp farming around the globe. Within shrimp populations WSSV is transmitted very fast, however, the modes and dynamics of transmission of this virus are not well understood. In the current study the dynamics of disease transmission of WSSV were investigated in small, closed populations of Penaeus monodon and Penaeus vannamei. Pair cohabitation experiments using PCR as a readout for virus infection were used to estimate transmission parameters for WSSV in these two species. The mortality rate of contact-infected shrimp in P. monodon was higher than the rate in P. vannamei. The transmission rate parameters for WSSV were not different between the two species. The relative contribution of direct and indirect transmission rates of WSSV differed between the two species. For P. vannamei the direct contact transmission rate of WSSV was significantly lower than the indirect environmental transmission rate, but for P. monodon, the opposite was found. The reproduction ratio R0 for WSSV for these two species of shrimp was estimated to be above one: 2.07 (95%CI 1.53, 2.79) for P. monodon and 1.51 (95%CI 1.12, 2.03) for P. vannamei. The difference in R0 between the two species is due to a lower host mortality and hence a longer infectious period of WSSV in P. monodon. Copyright © 2014 The Authors. Published by Elsevier B.V. All rights reserved.

  11. Thrips developmental stage-specific transcriptome response to tomato spotted wilt virus during the virus infection cycle in Frankliniella occidentalis, the primary vector.

    Science.gov (United States)

    Schneweis, Derek J; Whitfield, Anna E; Rotenberg, Dorith

    2017-01-01

    Tomato spotted wilt virus (TSWV) is transmitted by Frankliniella occidentalis in a circulative-propagative manner. Little is known about thrips vector response to TSWV during the infection process from larval acquisition to adult inoculation of plants. Whole-body transcriptome response to virus infection was determined for first-instar larval, pre-pupal and adult thrips using RNA-Seq. TSWV responsive genes were identified using preliminary sequence of a draft genome of F. occidentalis as a reference and three developmental-stage transcriptomes were assembled. Processes and functions associated with host defense, insect cuticle structure and development, metabolism and transport were perturbed by TSWV infection as inferred by ontologies of responsive genes. The repertoire of genes responsive to TSWV varied between developmental stages, possibly reflecting the link between thrips development and the virus dissemination route in the vector. This study provides the foundation for exploration of tissue-specific expression in response to TSWV and functional analysis of thrips gene function. Copyright © 2016 Elsevier Inc. All rights reserved.

  12. CONTROL OF VIRAL DISEASES TRANSMITTED IN A PERSISTENT MANNER BY THRIPS IN PEPPER (TOMATO SPOTTED WILT VIRUS).

    Science.gov (United States)

    Fanigliulo, A; Viggiano, A; Gualco, A; Crescenzi, A

    2014-01-01

    Tomato spotted wilt disease is caused by Tomato Spotted Wilt Virus (TSWV) (Tospovirus, Bunyaviridae), a virus that severely damages and reduces the yield of many economically important plants worldwide and actually it is a major disease affecting the production of tomato and pepper in Italy. Due to the non-predictive nature of its outbreaks combined with the lack of forecasting, adoption of preventive measures have not always been practical, in fact the disease cycle has proven to be extremely difficult to break because of the wide and often overlapping host range of both the virus and the thrips vectors, which transmit the virus in a persistent, circulative, and propagative manner. Moreover recently, resistance breaking (RB) isolates of TSWV that overcome the resistance conferred by the Tsw gene in different pepper hybrids have been recovered in different locations in Italy and also in Brazil, USA, Spain and Australia, and this occurrence raises the question on the importance of a new approach of integrated pest management for TSWV management, including both control of its insect vector and the induction of the plant's resistance against viral infection. In this perspective, a study was performed in 2012 and 2013 with the purpose of evaluating the efficacy of the insecticide Cyantraniliprole alone or combined with Acibenzolar-S-Methyl (ASM), inducer of systemic acquired resistance, in the control of tomato spotted wilt disease in pepper. The experiment was performed in laboratory, in a thermo-conditioned greenhouse, into separate insect-proof cages and consisted of 5 treatments and 2 applications (plus a pre-transplant application for treatments were ASM was used. Variables were the mode of application of ASM in pre-transplant (by foliar or by drench) and the duration of the exposure time of the treated plants to viruliferous insects. Pepper cv. Corno di Toro, devoid of any resistance to TSWV, was used. Plants were observed daily to record any symptom induced by

  13. Shrimp miR-12 Suppresses White Spot Syndrome Virus Infection by Synchronously Triggering Antiviral Phagocytosis and Apoptosis Pathways

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    Le Shu

    2017-07-01

    Full Text Available Growing evidence has indicated that the innate immune system can be regulated by microRNAs (miRNAs. However, the mechanism underlying miRNA-mediated simultaneous activation of multiple immune pathways remains unknown. To address this issue, the role of host miR-12 in shrimp (Marsupenaeus japonicus antiviral immune responses was characterized in the present study. The results indicated that miR-12 participated in virus infection, host phagocytosis, and apoptosis in defense against white spot syndrome virus invasion. miR-12 could simultaneously trigger phagocytosis, apoptosis, and antiviral immunity through the synchronous downregulation of the expression of shrimp genes [PTEN (phosphatase and tensin homolog and BI-1(transmembrane BAX inhibitor motif containing 6] and the viral gene (wsv024. Further analysis showed that miR-12 could synchronously mediate the 5′–3′ exonucleolytic degradation of its target mRNAs, and this degradation terminated in the vicinity of the 3′ untranslated region sequence complementary to the seed sequence of miR-12. Therefore, the present study showed novel aspects of the miRNA-mediated simultaneous regulation of multiple immune pathways.

  14. Dampak Stres Salinitas Terhadap Prevalensi White Spot Syndrome Virus (WSSV dan Survival Rate Udang Vannamei (Litopenaeus vannamei Pada Kondisi Terkontrol

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    Attabik Mukhammad Amrillah

    2015-08-01

    Full Text Available White spot syndrome (WSS adalah penyakit yang secara signifikan menyebabkan tingginya mortalitas dan kerusakan parah pada budidaya udang. Penelitian ini bertujuan mengetahui dampak stres salinitas terhadap prevalensi WSSV dan survival rate udang vannamei (Litopenaeus vannamei. Penelitian ini menggunakan udang vannamei ukuran PL 40 yang diinfeksi WSSV dengan konsentrasi virus 20 μg/ml pada tiga rentang salinitas yang berbeda 0-10 ppt, 11-20 ppt, 21–30 ppt dan di rendam selama 4 jam kemudian dilakukan pengamatan selama 7 hari pasca infeksi dan diukur survival rate dan kuaitas airnya. Hasil pengamatan menunjukkan bahwa seluruh sampel terinfeksi oleh WSSV, ditunjukkan hasil analisa PCR dan gejala klinis yang timbul. Salinitas 0-10 ppt memberikan hasil persentase survival rate terendah jika dibandingkan dengan rentang salinitas yang lainnya yaitu sebesar 7 ekor atau 33% dari jumlah total sampel yang digunakan. Persentase survival rate udang tertinggi pasca infeksi virus WSSV terdapat pada perlakuan salinitas 21-30 ppt yaitu sebesar 13 ekor atau 63% dari jumlah total individu, dan salinitas 11-20 ppt memiliki persentase survival rate medium yaitu sebesar 10 ekor atau 49% dari jumlah total sampel. Stres salinitas mempengaruhi prevalensi WSSV dengan semakin tingginya tingkat infeksi seiring menurunnya rentang salinitas, akan tetapi survival rate semakin tinggi seiring dengan bertambahnya rentang salinitas.

  15. Diversity of Thrips Species and Vectors of Tomato Spotted Wilt Virus in Tomato Production Systems in Kenya.

    Science.gov (United States)

    Macharia, Isaac; Backhouse, David; Skilton, Rob; Ateka, Elijah; Wu, Shu-Biao; Njahira, Moses; Maina, Solomon; Harvey, Jagger

    2015-02-01

    Thrips have been recognized as primary vectors of tomato spotted wilt virus (TSWV) with Frankliniella occidentalis (Pergande) reported as the most important and efficient vector, while other species such as Thrips tabaci Lindeman also include populations that can vector the virus. A study was undertaken to establish the diversity of thrips and presence of vectors for TSWV in four major tomato production areas in Kenya. The cytochrome oxidase 1 (CO1) gene was used to generate sequences from thrips samples collected from tomatoes and weeds, and phylogenetic analysis done to establish the variation within potential vector populations. Ceratothripoides brunneus Bagnall was the predominant species of thrips in all areas. F. occidentalis and T. tabaci were abundant in Nakuru, Kirinyaga, and Loitokitok but not detected at Bungoma. Other vectors of tospoviruses identified in low numbers were Frankliniella schultzei (Trybom) and Scirtothrips dorsalis Hood. Variation was observed in T. tabaci, F. occidentalis, and F. schultzei. Kenyan specimens of T. tabaci from tomato belonged to the arrhenotokous group, while those of F. occidentalis clustered with the Western flower thrips G group. The detection of RNA of TSWV in both of these species of thrips supported the role they play as vectors. The study has demonstrated the high diversity of thrips species in tomato production and the occurrence of important vectors of TSWV and other tospoviruses. © The Authors 2015. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

  16. An Elegant Analysis of White Spot Syndrome Virus Using a Graphene Oxide/Methylene Blue based Electrochemical Immunosensor Platform

    Science.gov (United States)

    Natarajan, Anusha; Devi, K. S. Shalini; Raja, Sudhakaran; Senthil Kumar, Annamalai

    2017-04-01

    White spot syndrome virus (WSSV) is a major devastating virus in aquaculture industry. A sensitive and selective diagnostic method for WSSV is a pressing need for the early detection and protection of the aquaculture farms. Herein, we first report, a simple electrochemical immunosensor based on methylene blue dye (MB) immobilized graphene oxide modified glassy carbon electrode (GCE/GO@MB) for selective, quick (35 ± 5 mins) and raw sample analysis of WSSV. The immunosensor was prepared by sequential modification of primary antibody, blocking agent (bovine serum album), antigen (as vp28 protein), secondary antibody coupled with horseradish peroxidase (Ab2-HRP) on the GCE/GO@MB. The modified electrode showed a well-defined redox peak at an equilibrium potential (E1/2), -0.4 V vs Ag/AgCl and mediated H2O2 reduction reaction without any false positive result and dissolved oxygen interferences in pH 7 phosphate buffer solution. Under an optimal condition, constructed calibration plot was linear in a range of 1.36 × 10-3 to 1.36 × 107 copies μL-1 of vp28. It is about four orders higher sensitive than that of the values observed with polymerase chain reaction (PCR) and western blot based WSSV detection techniques. Direct electrochemical immunosensing of WSSV in raw tissue samples were successfully demonstrated as a real sample system.

  17. Effect of Spinosad Resistance on Transmission of Tomato Spotted Wilt Virus by the Western Flower Thrips (Thysanoptera: Thripidae).

    Science.gov (United States)

    Zhao, Weiwei; Wan, Yanran; Xie, Wen; Xu, Baoyun; Zhang, Youjun; Wang, Shaoli; Wei, Guoshu; Zhou, Xiaomao; Wu, Qingjun

    2016-02-01

    Tomato spotted wilt virus (TSWV) is transmitted by Frankliniella occidentalis (Pergande) in a persistent-propagative manner. We previously observed significant results in terms of feeding behavior of spinosad-susceptible (Ivf03) and -resistant (Spin-R) strains of F. occidentalis using electrical penetration graph. TSWV transmission by the two strains was compared in the present study. The results showed that the titer of TSWV-N RNA (a part of S RNA of TSWV and encoding the nucleocapsid protein) in Ivf03 and Spin-R strains was not significantly different after a 48-h inoculation access period. The TSWV transmission rate did not significantly differ between the two strains and was 51.0% for Ivf03 and 44.4% for Spin-R. The virus transmission rate was significantly higher for males than females of both strains. The virus transmission rate for males and females of Ivf03 was 68.1 and 33.8%, respectively; however, in case of Spin-R, it was 60 and 28.8% for males and females, respectively. Additionally, number of probes and duration of probes were generally greater for viruliferous females of Ivf03 than for viruliferous females of Spin-R but the total number and duration of noningestion probes did not significantly differ between males of the two strains. The latter finding behavior may help explain the similar transmission rates for the susceptible and resistant strains. © The Authors 2015. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

  18. A Novel Vascular Endothelial Growth Factor Receptor Participates in White Spot Syndrome Virus Infection in Litopenaeus vannamei

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    Shihao Li

    2017-11-01

    Full Text Available Vascular endothelial growth factor (VEGF signaling pathway is known to play key roles in endothelial cell proliferation, migration, angiogenesis, vascular permeability, inhibition of apoptosis, and virus infection. In the present study, a novel VEGFR gene (LvVEGFR2 was identified and characterized from Litopenaeus vannamei. The deduced amino acid sequence of LvVEGFR2 possessed typical features of VEGFRs reported in other species, including six IG-like domains, a transmembrane motif, a protein kinase (PK domain, and one tyrosine-PK active site. The transcripts of LvVEGFR2 were mainly detected in hemocytes and lymphoid organ (Oka. Subcellular localization analysis showed that LvVEGFR2 was a membrane protein. Its expression level was obviously upregulated in hemocytes and Oka of the shrimp after white spot syndrome virus (WSSV infection. Knockdown of LvVEGFR2 gene expression by double-strand RNA mediated interference could lead to a decrease of virus copy number in WSSV-infected shrimp. The interaction between LvVEGFR2 and different LvVEGFs (LvVEGF1, LvVEGF2, and LvVEGF3 in shrimp was analyzed at the transcription level and protein level, respectively. Knockdown of LvVEGF2 or LvVEGF3 could downregulate the expression level of LvVEGFR2, and injection of the recombinant LvVEGF2 or LvVEGF3 could upregulate the expression level of LvVEGFR2. Yeast two-hybrid analysis showed that LvVEGFR2 could interact with LvVEGF2 and LvVEGF3 directly. The study improved our understanding on the VEGF signaling pathway of shrimp and its role during WSSV infection.

  19. Hemolymph cells apoptosis in imported shrimp Litopenaeus vannamei from Hawaii to Iran, exposed to white spot virus

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    Zeliha Selamoglu Talas

    2014-07-01

    Full Text Available Objective: To show hemolymph apoptosis in imported shrimp Litopenaeus vannamei from Hawaii to Iran. Methods: One hundred and eighty shrimps [(7.98±0.54 g] which were collected from a research shrimp farm located in Heleh site in north of Bushehr Province were distributed equally to 6 glass aquariums (50 cm×50 cm×60 cm as group A in triplicate (imported batch in 2011, without crossing with other generations with well clean aerated sea water (100 L per aquarium, salinity of 40 ‰ and temperature of 29 °C. Shrimps of group B (produced by crossing the adults of imported batches in 2009 up to 2011 were distributed also among 6 aquariums with the same conditions. Both shrimp groups were injected with concentration of LD50=1×10 5.4 white spot virus. Results: The results showed that in group A, the mortality began approximately 24 h after exposure and reached 100% after 36 h but no mortality was occurred up to 15 d in shrimps of group B. The slide evaluation of hemolymph of group B showed an increasing trend of apoptosis occurrence in all three types of hemolymph cells, hyalinocytes, semi-granulocytes and granulocytes from 24 h to 72 h in contrary to group A that not any apoptosis was observed during the course of the study (15 d. Conclusions: It is concluded that crossing among the specific pathogen free generations could induce the increasing immunity level through apoptosis to protect them against white spot disease.

  20. Outros constituintes isolados de Licania arianeae (Chrysobalanaceae

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    Mário G. de Carvalho

    Full Text Available O ácido flavona-6-sulfônico, 4'-O-metil-5,7-diidroxi-flavona-6-sulfonato, conhecido como niruriflavona, e a saponina, ácido 3-O-[6'-O-4-hidroxibenzoil]-²-D-galactopiranosil-ursa-12-en-28-óico, foram isolados, respectivamente, de madeira e folhas de Licania arianeae. As estruturas foram estabelecidas através da análise de espectros de massas e RMN incluindo experimentos bidimensionais.

  1. The Mechanism of Synchronous Precise Regulation of Two Shrimp White Spot Syndrome Virus Targets by a Viral MicroRNA

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    Yaodong He

    2017-11-01

    Full Text Available MicroRNAs (miRNAs, important factors in animal innate immunity, suppress the expressions of their target genes by binding to target mRNA’s 3′ untranslated regions (3′UTRs. However, the mechanism of synchronous regulation of multiple targets by a single miRNA remains unclear. In this study, the interaction between a white spot syndrome virus (WSSV miRNA (WSSV-miR-N32 and its two viral targets (wsv459 and wsv322 was characterized in WSSV-infected shrimp. The outcomes indicated that WSSV-encoded miRNA (WSSV-miR-N32 significantly inhibited virus infection by simultaneously targeting wsv459 and wsv322. The silencing of wsv459 or wsv322 by siRNA led to significant decrease of WSSV copies in shrimp, showing that the two viral genes were required for WSSV infection. WSSV-miR-N32 could mediate 5′–3′ exonucleolytic digestion of its target mRNAs, which stopped at the sites of target mRNA 3′UTRs close to the sequence complementary to the miRNA seed sequence. The complementary bases (to the target mRNA sequence of a miRNA 9th–18th non-seed sequence were essential for the miRNA targeting. Therefore, our findings presented novel insights into the mechanism of miRNA-mediated suppression of target gene expressions, which would be helpful for understanding the roles of miRNAs in innate immunity of invertebrate.

  2. Modified expression of alternative oxidase in transgenic tomato and petunia affects the level of tomato spotted wilt virus resistance

    Directory of Open Access Journals (Sweden)

    Ma Hao

    2011-10-01

    Full Text Available Abstract Background Tomato spotted wilt virus (TSWV has a very wide host range, and is transmitted in a persistent manner by several species of thrips. These characteristics make this virus difficult to control. We show here that the over-expression of the mitochondrial alternative oxidase (AOX in tomato and petunia is related to TSWV resistance. Results The open reading frame and full-length sequence of the tomato AOX gene LeAox1au were cloned and introduced into tomato 'Healani' and petunia 'Sheer Madness' using Agrobacterium-mediated transformation. Highly expressed AOX transgenic tomato and petunia plants were selfed and transgenic R1 seedlings from 10 tomato lines and 12 petunia lines were used for bioassay. For each assayed line, 22 to 32 tomato R1 progeny in three replications and 39 to 128 petunia progeny in 13 replications were challenged with TSWV. Enzyme-Linked Immunosorbent Assays showed that the TSWV levels in transgenic tomato line FKT4-1 was significantly lower than that of wild-type controls after challenge with TSWV. In addition, transgenic petunia line FKP10 showed significantly less lesion number and smaller lesion size than non-transgenic controls after inoculation by TSWV. Conclusion In all assayed transgenic tomato lines, a higher percentage of transgenic progeny had lower TSWV levels than non-transgenic plants after challenge with TSWV, and the significantly increased resistant levels of tomato and petunia lines identified in this study indicate that altered expression levels of AOX in tomato and petunia can affect the levels of TSWV resistance.

  3. Evaluation and identification of candidate genes for artificial microRNA-mediated resistance to tomato spotted wilt virus.

    Science.gov (United States)

    Mitter, Neena; Zhai, Ying; Bai, Anh Xu; Chua, Keith; Eid, Sahar; Constantin, Myrna; Mitchell, Roger; Pappu, Hanu R

    2016-01-04

    Tomato spotted wilt virus (TSWV) is an economically important viral pathogen of a wide range of field and horticultural crops. We developed an artificial microRNA (amiRNA) strategy against TSWV, targeting the nucleoprotein (N) and silencing suppressor (NSs) genes. The amiRNA constructs replaced the natural miRNA in a shortened Arabidopsis 173-nucleotide (nt) miR159a precursor backbone (athmiR159a) without the stem base extending beyond the miR/miR* duplex. Further, each amiRNA was modified to contain a mismatch (wobble) sequence at nucleotide position 12 and 13 on the complementary strand amiRNA*, mimicking the endogenous miR159a sequence structure. Transient expression in Nicotiana benthamiana demonstrated that the introduction of a wobble sequence did not alter amiRNA expression levels. Following challenge inoculation with TSWV, plants expressing N-specific amiRNAs with or without the wobble remained asymptomatic and were negative for TSWV by ELISA. In contrast, plants expressing the NSs-specific amiRNAs were symptomatic and accumulated high levels of TSWV. Similar findings were obtained in stably transformed Nicotiana tabacum plants. Our results show that a shortened 173-nt athmiR159a backbone is sufficient to express amiRNAs and that the presence of mismatch at position 12-13 does not influence amiRNA expression or conferring of resistance. We also show that selection of target gene and positional effect are critical in amiRNA-based approach for introducing resistance. These findings open the possibility of employing the amiRNA approach for broad-spectrum resistance to tospoviruses as well as other viruses. Copyright © 2015 Elsevier B.V. All rights reserved.

  4. Development of Rapid Immune-gold Strip Kit for On-Site Diagnosis of Tomato spotted wilt virus

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    Ju-Yeon Yoon

    2014-03-01

    Full Text Available A rapid, user-friendly and simple immune-chromatographic dipstick kit named ‘rapid immune-gold strip’ (RIGS kit was developed in a novel single strip format to detect on-site detection of Tomato spotted wilt virus (TSWV. Immunoglobulin G (IgG from polyclonal antisera raised in rabbits against TSWV was purified through protein- A affinity chromatography and then the purified TSWV-IgG was conjugated to colloidal gold nano-particles which served as a test line on nitrocellulose membrane. Protein A that non-specifically binds to TSWV antibody was used as a control line on the same strip. The diagnosis process with the TSWV-RIGS involves simply grinding the suspect plant sample in a bag that contains the extraction buffer and inserting the strip the bag. Results can be seen in 2-5 minutes. The flow of the complexes of gold particles coated with TSWV-IgG and a crude sap from TSWV-infected pepper, tobacco and tomato plants resulted in intensive color formed on the test lines proportional to the concentrations of TSWV. The RIGS-TSWV kit did not show any cross-reactions against other tomato-infecting viruses unrelated to TSWV. These results indicate that the TSWV-RIGS kit is highly sensitive and is not required for laboratory training and experience prior to testing. The TSWV-RIGS kit is suitable for on-site detection of suspect TSWV-infected plants as well as for laboratory diagnosis.

  5. An Investigation into Occasional White Spot Syndrome Virus Outbreak in Traditional Paddy Cum Prawn Fields in India

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    Deborah Gnana Selvam

    2012-01-01

    Full Text Available A yearlong (September 2009–August 2010 study was undertaken to find out possible reasons for occasional occurrence of White Spot Syndrome Virus (WSSV outbreak in the traditional prawn farms adjoining Cochin backwaters. Physicochemical and bacteriological parameters of water and sediment from feeder canal and four shrimp farms were monitored on a fortnightly basis. The physicochemical parameters showed variation during the two production cycles and between the farms studied. Dissolved oxygen (DO content of water from feeder canal showed low oxygen levels (as low as 0.8 mg/L throughout the study period. There was no disease outbreak in the perennial ponds. Poor water exchange coupled with nutrient loading from adjacent houses resulted in phytoplankton bloom in shallow seasonal ponds which led to hypoxic conditions in early morning and supersaturation of DO in the afternoon besides considerably high alkaline pH. Ammonia levels were found to be very high in these ponds. WSSV outbreak was encountered twice during the study leading to mass mortalities in the seasonal ponds. The hypoxia and high ammonia content in water and abrupt fluctuations in temperature, salinity and pH might lead to considerable stress in the shrimps triggering WSSV infection in these traditional ponds.

  6. Evolution and structure of Tomato spotted wilt virus populations: evidence of extensive reassortment and insights into emergence processes.

    Science.gov (United States)

    Tentchev, Diana; Verdin, Eric; Marchal, Cécile; Jacquet, Monique; Aguilar, Juan M; Moury, Benoît

    2011-04-01

    Tomato spotted wilt virus (TSWV; genus Tospovirus, family Bunyaviridae) genetic diversity was evaluated by sequencing parts of the three RNA genome segments of 224 isolates, mostly from pepper and tomato crops in southern Europe. Eighty-three per cent of the isolates showed consistent clustering into three clades, corresponding to their geographical origin, Spain, France or the USA, for the three RNA segments. In contrast, the remaining 17% of isolates did not belong to the same clade for the three RNA segments and were shown to be reassortants. Among them, eight different reassortment patterns were observed. Further phylogenetic analyses provided insights into the dynamic processes of the worldwide resurgence of TSWV that, since the 1980s, has followed the worldwide dispersal of the western flower thrips (Frankliniella occidentalis) tospovirus vector. For two clades composed essentially of Old World (OW) isolates, tree topology suggested a local re-emergence of indigenous TSWV populations following F. occidentalis introductions, while it could not be excluded that the ancestors of two other OW clades were introduced from North America contemporarily with F. occidentalis. Finally, estimation of the selection intensity that has affected the evolution of the NSs and nucleocapsid proteins encoded by RNA S of TSWV suggests that the former could be involved in the breakdown of resistance conferred by the Tsw gene in pepper.

  7. Temporal dynamics of iris yellow spot virus and its vector, Thrips tabaci (Thysanoptera: Thripidae), in seeded and transplanted onion fields.

    Science.gov (United States)

    Hsu, Cynthia L; Hoepting, Christine A; Fuchs, Marc; Shelton, Anthony M; Nault, Brian A

    2010-04-01

    Onion thrips, Thrips tabaci (Lindeman) (Thysanoptera: Thripidae), can reduce onion bulb yield and transmit iris yellow spot virus (IYSV) (Bunyaviridae: Tospovirus), which can cause additional yield losses. In New York, onions are planted using seeds and imported transplants. IYSV is not seed transmitted, but infected transplants have been found in other U.S. states. Transplants are also larger than seeded onions early in the season, and thrips, some of which may be viruliferous, may preferentially colonize larger plants. Limited information is available on the temporal dynamics of IYSV and its vector in onion fields. In 2007 and 2008, T. tabaci and IYSV levels were monitored in six seeded and six transplanted fields. We found significantly more thrips in transplanted fields early in the season, but by the end of the season seeded fields had higher levels of IYSV. The percentage of sample sites with IYSV-infected plants remained low (thrips in August and September were better predictors of final IYSV levels than early season thrips densities. For 2007 and 2008, the time onions were harvested may have been more important in determining IYSV levels than whether the onions were seeded or transplanted. Viruliferous thrips emigrating from harvested onion fields into nonharvested ones may be increasing the primary spread of IYSV in late-harvested onions. Managing T. tabaci populations before harvest, and manipulating the spatial arrangement of fields based on harvest date could mitigate the spread of IYSV.

  8. Highly Sensitive Detection of Low-Abundance White Spot Syndrome Virus by a Pre-Amplification PCR Method.

    Science.gov (United States)

    Pan, Xiaoming; Zhang, Yanfang; Sha, Xuejiao; Wang, Jing; Li, Jing; Dong, Ping; Liang, Xingguo

    2017-03-28

    White spot syndrome virus (WSSV) is a major threat to the shrimp farming industry and so far there is no effective therapy for it, and thus early diagnostic of WSSV is of great importance. However, at the early stage of infection, the extremely low-abundance of WSSV DNA challenges the detection sensitivity and accuracy of PCR. To effectively detect low-abundance WSSV, here we developed a pre-amplification PCR (pre-amp PCR) method to amplify trace amounts of WSSV DNA from massive background genomic DNA. Combining with normal specific PCR, 10 copies of target WSSV genes were detected from ~1010 magnitude of backgrounds. In particular, multiple target genes were able to be balanced amplified with similar efficiency due to the usage of the universal primer. The efficiency of the pre-amp PCR was validated by nested-PCR and quantitative PCR, and pre-amp PCR showed higher efficiency than nested-PCR when multiple targets were detected. The developed method is particularly suitable for the super early diagnosis of WSSV, and has potential to be applied in other low-abundance sample detection cases.

  9. Novel Insights into Antiviral Gene Regulation of Red Swamp Crayfish, Procambarus clarkii, Infected with White Spot Syndrome Virus

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    Shaokui Yi

    2017-11-01

    Full Text Available White spot syndrome virus (WSSV, one of the major pathogens of Procambarus clarkii, has caused severe disruption to the aquaculture industry of P. clarkii in China. To reveal the gene regulatory mechanisms underlying WSSV infection, a comparative transcriptome analysis was performed among WSSV-infected susceptible individuals (GS, viral resistant individuals (GR, and a non-infected control group (GC. A total of 61,349 unigenes were assembled from nine libraries. Subsequently, 515 and 1033 unigenes exhibited significant differential expression in sensitive and resistant crayfish individuals compared to the control group (GC. Many differentially expressed genes (e.g., C-type lectin 4, Peroxinectin, Prophenoloxidase, and Serine/threonine-protein kinase observed in GR and GS play critical roles in pathogen recognition and viral defense reactions after WSSV infection. Importantly, the glycosaminoglycan biosynthesis-chondroitin sulfate/dermatan sulfate pathway was identified to play critical roles in defense to WSSV infection for resistant crayfish individuals by upregulating the chondroitin sulfate related genes for the synthesis of WSSV-sensitive, functional chondroitin sulfate chains containing E units. Numerous genes and the key pathways identified between resistant and susceptible P. clarkii individuals provide valuable insights regarding antiviral response mechanisms of decapoda species and may help to improve the selective breeding of P. clarkii WSSV-resistance.

  10. Molecular modeling and expression of the Litopenaeus vannamei proliferating cell nuclear antigen (PCNA) after white spot syndrome virus shrimp infection.

    Science.gov (United States)

    de-la-Re-Vega, Enrique; Muhlia-Almazan, Adriana; Arvizu-Flores, Aldo A; Islas-Osuna, Maria A; Yepiz-Plascencia, Gloria; Brieba, Luis G; Sotelo-Mundo, Rogerio R

    2011-01-01

    Proliferating cell nuclear antigen (PCNA) is the eukaryotic sliding clamp that tethers DNA polymerase to DNA during replication. The full-length cDNA of the Pacific white shrimp Litopenaeus vannamei PCNA (LvPCNA) was cloned and encoded a protein of 260 amino acids that is highly similar to other Crustacean PCNAs. The theoretical shrimp PCNA structure has all the domains that are necessary for its interaction with template DNA and DNA polymerase. RT-PCR analysis showed that LvPCNA is expressed mainly in muscle and hemocytes and much less in hepatopancreas and gills. LvPCNA mRNA levels are not statistically different in muscle from healthy and challenged shrimp with the white spot syndrome virus (WSSV). In contrast, the mRNA levels of the viral DNA polymerase show a biphasic pattern with expression at 6 h post-infection and later at 24 and 48 h. These results suggest that in shrimp muscle LvPCNA levels are steadily kept to allow viral replication and that WSSV DNA polymerase (WSSV-DNApol) is more responsive towards later stages of infection. More knowledge of the DNA replication machinery would result in a better understanding of the mechanism and components of viral replication, since the WSSV genome does not have all the components required for assembly of a fully functional replisome.

  11. Virulence of white spot syndrome virus (WSSV) isolates may be correlated with the degree of replication in gills of Penaeus vannamei juveniles

    OpenAIRE

    Rahman, M.M; Corteel, M.; Escobedo-Bonilla, C.M.; Wille, M.; Alday-Sanz, V.; Pensaert, M. B.; Sorgeloos, P.; Nauwynck, H J

    2008-01-01

    A standardized inoculation model was used in 2 separate experiments to gauge the virulence of 3 white spot syndrome virus (WSSV) isolates from Thailand and Vietnam (WSSV Thai-1, WSSV Thai-2, and WSSV Viet) in Penaeus vannamei juveniles. Mortality patterns (Expt 1) were compared and WSSV-positive cells quantified (Expt 2) in tissues following intramuscular inoculation of shrimp with the most (WSSV Thai-1) and least (WSSV Viet) virulent isolates as determined by Expt 1. The results of Expt 1 de...

  12. Resistance to tospoviruses in Nicotiana benthamiana transformed with the N gene of tomato spotted wilt virus: correlation between transgene expression and protection in primary transformants.

    Science.gov (United States)

    Vaira, A M; Semeria, L; Crespi, S; Lisa, V; Allavena, A; Accotto, G P

    1995-01-01

    Nicotiana benthamiana was transformed with the nucleoprotein (N) gene of an Italian isolate of tomato spotted wilt virus (TSWV). Forty-five T1 primary transformant lines were analyzed for the expression of N protein and for resistance to TSWV and three other tospoviruses: impatiens necrotic spot virus (INSV), groundnut bud necrosis virus (GBNV), and groundnut ringspot virus (GRSV). Thirteen of these lines were further characterized. Resistance to all TSWV isolates tested was found in two lines. The expression of the transgene (N mRNA) was lower in these resistant lines than in any of the susceptible lines, and the transgene N protein was either absent or present below detectable levels. These lines were susceptible to the other tospoviruses tested, but they developed symptoms milder than controls when inoculated with GRSV. Some of the lines producing high levels of N protein showed delays (of 2-3 weeks) in symptom expression with at least one of the TSWV isolates tested and symptom delay or attenuation with INSV or GRSV (or both). From our results it appears that high expression of TSWV N protein retards, in some cases, disease development by TSWV and INSV. In contrast, the lack of detectable expression of the transgenic N protein, accompanied by limited production of N transcripts, conferred TSWV-specific resistance.

  13. Potencial de antagonismo de isolados de Trichoderma sp. contra o isolados de Fusarium sp., in vitro

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    Charles Anthony Hoffmann

    2015-06-01

    Full Text Available Resumo – O uso discriminado de agrotóxicos acarreta em uma grande prejuízo a saúdes das pessoas e do meio ambiente. O uso de novos métodos de combate a pragas se faz necessário para o equilíbrio da cadeia produtiva. A utilização de microrganismos no combate a pragas se faz como uma boa alternativa. O trabalho objetivou-se em avaliar o potencial de antagonismo de isolados de Trichoderma sp. contra o patógeno Fusarium sp. in vitro. Para isso foi utilizado os métodos de confronto direto, pareamento de culturas e de metabólitos voláteis. Os experimentos foram realizados na incubadora de empresas da Universidade Federal do Tocantins – UFT Campus de Gurupi, seguiram com três repetições. Foram testados 15 isolados do antagonista em confronto direto com o patógeno e os mesmos isolados e patógeno foram testados em metabólitos voláteis ambos utilizando o meio de cultura BDA. Todos os isolados apresentaram antagonismo in vitro contra o patógeno.

  14. Transcriptome-wide identification of host genes targeted by tomato spotted wilt virus-derived small interfering RNAs.

    Science.gov (United States)

    Ramesh, Shunmugiah V; Williams, Sarah; Kappagantu, Madhu; Mitter, Neena; Pappu, Hanu R

    2017-06-15

    RNA silencing mechanism functions as a major defense against invading viruses. The caveat in the RNA silencing mechanism is that the effector small interfering RNAs (siRNAs) act on any RNA transcripts with sequence complementarity irrespective of target's origin. A subset of highly expressed viral small interfering RNAs (vsiRNAs) derived from the tomato spotted wilt virus (TSWV; Tospovirus: Bunyaviridae) genome was analyzed for their propensity to downregulate the tomato transcriptome. A total of 11898 putative target sites on tomato transcripts were found to exhibit a propensity for down regulation by TSWV-derived vsiRNAs. In total, 2450 unique vsiRNAs were found to have potential cross-reacting capability with the tomato transcriptome. VsiRNAs were found to potentially target a gamut of host genes involved in basal cellular activities including enzymes, transcription factors, membrane transporters, and cytoskeletal proteins. KEGG pathway annotation of targets revealed that the vsiRNAs were mapped to secondary metabolite biosynthesis, amino acids, starch and sucrose metabolism, and carbon and purine metabolism. Transcripts for protein processing, hormone signalling, and plant-pathogen interactions were the most likely targets from the genetic, environmental information processing, and organismal systems, respectively. qRT-PCR validation of target gene expression showed that none of the selected transcripts from tomato cv. Marglobe showed up regulation, and all were down regulated even upto 20 folds (high affinity glucose transporter). However, the expression levels of transcripts from cv. Red Defender revealed differential regulation as three among the target transcripts showed up regulation (Cc-nbs-lrr, resistance protein, AP2-like ethylene-responsive transcription factor, and heat stress transcription factor A3). Accumulation of tomato target mRNAs of corresponding length was proved in both tomato cultivars using 5' RACE analysis. The TSWV-tomato interaction at

  15. Salicylic Acid Is Involved in the Basal Resistance of Tomato Plants to Citrus Exocortis Viroid and Tomato Spotted Wilt Virus.

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    M Pilar López-Gresa

    Full Text Available Tomato plants expressing the NahG transgene, which prevents accumulation of endogenous salicylic acid (SA, were used to study the importance of the SA signalling pathway in basal defence against Citrus Exocortis Viroid (CEVd or Tomato Spotted Wilt Virus (TSWV. The lack of SA accumulation in the CEVd- or TSWV-infected NahG tomato plants led to an early and dramatic disease phenotype, as compared to that observed in the corresponding parental Money Maker. Addition of acibenzolar-S-methyl, a benzothiadiazole (BTH, which activates the systemic acquired resistance pathway downstream of SA signalling, improves resistance of NahG tomato plants to CEVd and TSWV. CEVd and TSWV inoculation induced the accumulation of the hydroxycinnamic amides p-coumaroyltyramine, feruloyltyramine, caffeoylputrescine, and feruloylputrescine, and the defence related proteins PR1 and P23 in NahG plants earlier and with more intensity than in Money Maker plants, indicating that SA is not essential for the induction of these plant defence metabolites and proteins. In addition, NahG plants produced very high levels of ethylene upon CEVd or TSWV infection when compared with infected Money Maker plants, indicating that the absence of SA produced additional effects on other metabolic pathways. This is the first report to show that SA is an important component of basal resistance of tomato plants to both CEVd and TSWV, indicating that SA-dependent defence mechanisms play a key role in limiting the severity of symptoms in CEVd- and TSWV-infected NahG tomato plants.

  16. Characterization of Tomato spotted wilt virus isolates that overcome the Sw-5 resistance gene in tomato and fitness assays

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    Jose ARAMBURU

    2011-01-01

    Full Text Available Resistance-breaking (RB isolates of Tomato spotted wilt virus (TSWV that overcome the resistance conferred by the Sw-5 gene in tomato have had only a limited spread since they were first detected in north-eastern Spain in 2002. Symptom expression, homogeneity, stability and the transmission capacity of RB and non-resistance breaking (NRB isolates were biologically compared. The fitness of both types of isolates infecting tomato plants was determined in competition assays. All TSWV isolates induced similar systemic symptoms in a wide range of plant species, except RB isolates in tomato carrying the Sw-5 resistance gene and pepper carrying the Tsw resistance gene. The mechanical transmission of RB isolates to tomato plants with the Sw-5 gene failed in some trials, although NRB isolates did not differ noticeably in transmission efficiency when tested with the thrips Frankliniella occidentalis. Biological clones from individual local lesions obtained by mechanically inoculating Nicotiana glutinosa in some TSWV field samples showed that they were biologically homogeneous. Mixed infections of RB and wilt-type isolates were not found. The RB isolates were relatively stable because no reversion to NRB isolates was seen after serial passages in susceptible tomato plants. In competition assays between RB and NRB isolates, after serial passages in susceptible tomato plants, the prevalence of a particular isolate was not related to its capacity to overcome Sw-5 gene resistance. The low spread of the RB isolates in Spain does not seem to be related to a loss of fitness in tomato plants or to differences in transmission capacity by thrips, but it could be related to the reduction of the selection pressure of RB isolates as consequence of the gradual replacement of susceptible tomato plants by resistant tomato plants by growers.

  17. Salicylic Acid Is Involved in the Basal Resistance of Tomato Plants to Citrus Exocortis Viroid and Tomato Spotted Wilt Virus.

    Science.gov (United States)

    López-Gresa, M Pilar; Lisón, Purificación; Yenush, Lynne; Conejero, Vicente; Rodrigo, Ismael; Bellés, José María

    2016-01-01

    Tomato plants expressing the NahG transgene, which prevents accumulation of endogenous salicylic acid (SA), were used to study the importance of the SA signalling pathway in basal defence against Citrus Exocortis Viroid (CEVd) or Tomato Spotted Wilt Virus (TSWV). The lack of SA accumulation in the CEVd- or TSWV-infected NahG tomato plants led to an early and dramatic disease phenotype, as compared to that observed in the corresponding parental Money Maker. Addition of acibenzolar-S-methyl, a benzothiadiazole (BTH), which activates the systemic acquired resistance pathway downstream of SA signalling, improves resistance of NahG tomato plants to CEVd and TSWV. CEVd and TSWV inoculation induced the accumulation of the hydroxycinnamic amides p-coumaroyltyramine, feruloyltyramine, caffeoylputrescine, and feruloylputrescine, and the defence related proteins PR1 and P23 in NahG plants earlier and with more intensity than in Money Maker plants, indicating that SA is not essential for the induction of these plant defence metabolites and proteins. In addition, NahG plants produced very high levels of ethylene upon CEVd or TSWV infection when compared with infected Money Maker plants, indicating that the absence of SA produced additional effects on other metabolic pathways. This is the first report to show that SA is an important component of basal resistance of tomato plants to both CEVd and TSWV, indicating that SA-dependent defence mechanisms play a key role in limiting the severity of symptoms in CEVd- and TSWV-infected NahG tomato plants.

  18. Effects of white spot syndrome virus infection on immuno-enzyme activities and ultrastructure in gills of Cherax quadricarinatus.

    Science.gov (United States)

    Wang, Dan-Li; Zuo, Di; Wang, Lan-Mei; Sun, Ting; Wang, Qun; Zhao, Yun-Long

    2012-05-01

    In this study, we explored the pathogenic mechanism of white spot syndrome virus (WSSV) in crayfish, Cherax quadricarinatus, by investigating activities of enzymes related to innate immune function during infection. After 6-12 h of exposure to WSSV, the activities of four enzymes, phenoloxidase (PO), peroxidase (POD), superoxide dismutase (SOD) and lysozyme (LSZ), increased in the gills of C. quadricarinatus but then sharply decreased during longer infection times. Except for PO, the activities of other enzymes in the WSSV-infected crayfish (Group II) were significantly lower than those of the controls at 72 h post-exposure (P < 0.01). Interestingly, the enzyme activities in the group treated with polysaccharides before challenge with WSSV (Group III) were higher than those in Group II. This phenomenon demonstrated that the polysaccharides could improve the immuno-enzyme activities and enhance the organism's antiviral defenses. Morphological examination by transmission electron microscopy revealed abundant WSSV particles and significant damage in the gills of infected crayfish. WSSV infection caused parts of the gill epithelium and microvilli to be reduced in number and size or damaged; meanwhile, the mitochondria morphology changed, with parts of the cristae diminished leaving large vacuoles. Moreover, electron dense deposits appeared and heterochromatinized nuclei could be seen in blood cells with ruptured nuclear membranes and outflow of nucleoplasm. The findings of this study furthers our understanding of the biochemical alterations induced by viral infections, including changes in the antioxidant status, oxidative stress and lysozyme activity, which could help to advance strategies for control of WSSV in crayfish. Copyright © 2012 Elsevier Ltd. All rights reserved.

  19. [Expression of single chain fragment variable P1D3 antibody against shrimp white spot syndrome virus in Pichia pastoris].

    Science.gov (United States)

    Yang, Yi; Zhang, Min; Yuan, Li; Zhang, Xiao-Hua; Dai, He-Ping

    2006-11-01

    White spot syndrome virus (WSSV) is a major pathogen in aquaculture penaeid shrimp, which caused catastrophic economic losses in the worldwide. No adequate treatments against WSSV are available. In order to study infection mechanism of WSSV, a phage display scFv cDNA library against WSSV was constructed and a neutralizing antibody of scFv P1D3 was selected in our lab previously. In this study, scFv P1D3 was expressed successfully in yeast. Firstly, the original expression vector of P1D3, M13 phagmid, was used as a template to design primers with restriction sites of SnaB I and EcoR I . Then the gene of P1D3 was amplified by PCR. After digested by SnaB I and EcoR I , the fragment of scFv P1D3 with E-tag was inserted into yeast and E. coli shuttle plasmid pPIC9k. The recombinant plasmid pPIC9k-scFv P1D3-Etag was linearized with Bgl II and then transformed into Pichia pastoris GS115 by electroporation. Positive clones were selected and verified by PCR and DNA sequencing. The scFv PID3 was induced to express in yeast by methanol. The results of ELISA demonstrate that scFv P1D3 expressed in yeast still has high specificity to bind on WSSV and the binding activity is higher than that expressed in E. coli TG1. After several optimizing experiments, the results show that the expression amount of scFv P1D3 can reach to 302 mg/L in yeast culture supernatant. This experiment has offered a new source of antibody for the researches on passive immunology for shrimp.

  20. Impact of Vibrio parahaemolyticus and white spot syndrome virus (WSSV) co-infection on survival of penaeid shrimp Litopenaeus vannamei

    Science.gov (United States)

    Zhang, Xiaojing; Song, Xiaoling; Huang, Jie

    2016-11-01

    White spot syndrome virus (WSSV) is an important viral pathogen that infects farmed penaeid shrimp, and the threat of Vibrio parahaemolyticus infection to shrimp farming has become increasingly severe. Viral and bacterial cross or superimposed infections may induce higher shrimp mortality. We used a feeding method to infect Litopenaeus vannamei with WSSV and then injected a low dose of V. parahaemolyticus (WSSV+Vp), or we first infected L. vannamei with a low-dose injection of V. parahaemolyticus and then fed the shrimp WSSV to achieve viral infection (Vp+WSSV). The eff ect of V. parahaemolyticus and WSSV co-infection on survival of L. vannamei was evaluated by comparing cumulative mortality rates between experimental and control groups. We also spread L. vannamei hemolymph on thiosulfate citrate bile salt sucrose agar plates to determine the number of Vibrio, and the WSSV copy number in L. vannamei gills was determined using an absolute quantitative polymerase chain reaction (PCR) method. LvMyD88 and Lvakt gene expression levels were detected in gills of L. vannamei by real-time PCR to determine the cause of the diff erent mortality rates. Our results show that (1) the cumulative mortality rate of L. vannamei in the WSSV+Vp group reached 100% on day 10 after WSSV infection, whereas the cumulative mortality rate of L. vannamei in the Vp+WSSV group and the WSSV-alone control group approached 100% on days 11 and 13 of infection; (2) the number of Vibrio in the L. vannamei group infected with V. parahaemolyticus alone declined gradually, whereas the other groups showed significant increases in the numbers of Vibrio ( Pvannamei and vice versa. The combined accelerated proliferation of both V. parahaemolyticus and WSSV led to massive death of L. vannamei.

  1. Influence of white spot syndrome virus infection on hepatopancreas gene expression of `Huanghai No. 2' shrimp ( Fenneropenaeus chinensis)

    Science.gov (United States)

    Meng, Xianhong; Shi, Xiaoli; Kong, Jie; Luan, Sheng; Luo, Kun; Cao, Baoxiang; Liu, Ning; Lu, Xia; Li, Xupeng; Deng, Kangyu; Cao, Jiawang; Zhang, Yingxue; Zhang, Hengheng

    2017-10-01

    To elucidate the molecular response of shrimp hepatopancreas to white spot syndrome virus (WSSV) infection, microarray was applied to investigate the differentially expressed genes in the hepatopancreas of `Huanghai No. 2' ( Fenneropenaeus chinensis). A total of 59137 unigenes were designed onto a custom-made 60K Agilent chip. After infection, the gene expression profiles in the hepatopancreas of the shrimp with a lower viral load at early (48-96 h), peak (168-192 h) and late (264-288 h) infection phases were analyzed. Of 18704 differentially expressed genes, 6412 were annotated. In total, 5453 differentially expressed genes (1916 annotated) expressed at all three phases, and most of the annotated were either up- or down-regulated continuously. These genes function diversely in, for example, immune response, cytoskeletal system, signal transduction, stress resistance, protein synthesis and processing, metabolism among others. Some of the immune-related genes, including antilipopolysaccharide factor, Kazal-type proteinase inhibitor, C-type lectin and serine protease encoding genes, were up-regulated after WSSV infection. These genes have been reported to be involved in the anti-WSSV responses. The expression of genes related to the cytoskeletal system, including β-actin and myosin but without tubulin genes, were down-regulated after WSSV infection. Astakine was found for the first time in the WSSV-infected F. chinensis. To further confirm the expression of differentially expressed genes, quantitative real-time PCR was performed to test the expression of eight randomly selected genes and verified the reliability and accuracy of the microarray expression analysis. The data will provide valuable information to understanding the immune mechanism of shrimp's response to WSSV.

  2. Comparison of white spot syndrome virus infection resistance between Exopalaemon carinicauda and Litopenaeus vannamei under different salinity stresses

    Science.gov (United States)

    Ge, Qianqian; Yu, Ge; Sun, Ming; Li, Jitao; Li, Jian

    2017-12-01

    Exopalaemon carinicauda is one of the important economic shrimp species in China, and can tolerate a wide range of salinities. However, its disease resistance remains to be unclear in comparison with other shrimp species under salinity stress. In this study, the resistance to white spot syndrome virus (WSSV) of E. carinicauda and Litopenaeus vannamei was determined by comparing their hemocyanin (Hc) and phenoloxidase (PO) activities under different salinity stresses. In E. carinicauda, the PO activity and Hc gene transcript abundance showed a coherent pattern of increase and decrease while Hc content showed a slightly decrease with Vibrio anguillarum and WSSV infections. For both E. carinicauda and L. vannamei under salinity stress, the PO activity showed a positive correlation with the salinity while the Hc content and expression level of its gene increased significantly in salinities of 5, 15 and 25 g L-1. The survival rate of E. carinicauda with WSSV infection was higher than that of L. vannamei in the first 24 h under different salinity stresses. Drastic mortality of E.carinicauda and L. vannamei appeared at 48 h and 3 h post-injection, respectively. Furthermore, compared with L. vannamei, E. carinicauda displayed higher PO activity, Hc content and abundance of Hc gene mRNA. The results collectively indicated that Hc and PO have obviously functional connection in resisting pathogens and tolerating salinity stress, and PO activity and Hc gene mRNA abundance may reflect the resistance of shrimp to disease. E. carinicauda has higher level of immune potential than L. vannamei, suggesting its greater capacity in resisting pathogens under salinity stresses.

  3. Patogenicidade de isolados de Corynespora cassiicola a diferentes espécies de plantas Pathogenicity of Corynespora cassiicola isolates on different host plants

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    Ricardo Ribeiro Oliveira

    2007-09-01

    Full Text Available Corynespora cassiicola, relatado como um patógeno com ampla gama de espécies hospedeiras, tem causado danos em várias culturas de interesse comercial. Neste trabalho avaliou-se, em ambiente de casa-de-vegetação, a patogenicidade de 15 isolados de C. cassiicola originados de diferentes espécies hospedeiras, quando inoculados em 12 diferentes espécies vegetais. Os isolados de pepino foram os mais patogênicos. Além dos híbridos de pepino, estes infectaram outras seis espécies vegetais testadas. Já os isolados de trapoeraba e de alface foram os que apresentaram menor patogenicidade, pois além dos hospedeiros originais infectaram somente uma espécie hospedeira. A maioria dos isolados apresentou elevada inespecificidade. As espécies vegetais testadas reagiram de formas diferentes quando inoculadas com diferentes isolados. O mamoeiro apresentou maior suscetibilidade, sendo infectado por 12 dos 15 isolados. Contrariamente, a trapoeraba e o assa-peixe foram suscetíveis a três e dois isolados, respectivamente.Corynespora cassiicola, causes target leaf spot on a wide host range, including several economically important crops. This study investigated the pathogenicity of 15 C. cassiicola isolates on several hosts, in greenhouse. The fungal isolates were collected from several hosts. These isolates were inoculated on 12 different plant species. The isolates from cucumber showed the widest host range, infecting also six other hosts. The isolates from C. benghalensis and lettuce showed the narrowest host range, since they infected their host of origin and only another host plant. Most of the isolates lacked host specificity. Papaya plant, which showed the greatest susceptibility to the C. cassiicola isolates, were colonized by 12 of the 15 isolates tested. On the opposite, Vernonia sp. and C benghalensis were susceptible to only two and three C. cassiicola isolates, respectively.

  4. A soluble form of the Tomato spotted wilt virus (TSWV) glycoprotein G(N) (G(N)-S) inhibits transmission of TSWV by Frankliniella occidentalis.

    Science.gov (United States)

    Whitfield, A E; Kumar, N K K; Rotenberg, D; Ullman, D E; Wyman, E A; Zietlow, C; Willis, D K; German, T L

    2008-01-01

    Tomato spotted wilt virus (TSWV) is an economically important virus that is transmitted in a persistent propagative manner by its thrips vector, Frankliniella occidentalis. Previously, we found that a soluble form of the envelope glycoprotein G(N) (G(N)-S) specifically bound thrips midguts and reduced the amount of detectable virus inside midgut tissues. The aim of this research was to (i) determine if G(N)-S alters TSWV transmission by thrips and, if so, (ii) determine the duration of this effect. In one study, insects were given an acquisition access period (AAP) with G(N)-S mixed with purified virus and individual insects were assayed for transmission. We found that G(N)-S reduced the percent of transmitting adults by eightfold. In a second study, thrips were given an AAP on G(N)-S protein and then placed on TSWV-infected plant material. Individual insects were assayed for transmission over three time intervals of 2 to 3, 4 to 5, and 6 to 7 days post-adult eclosion. We observed a significant reduction in virus transmission that persisted to the same degree throughout the time course. Real-time reverse transcription polymerase chain reaction analysis of virus titer in individual insects revealed that the proportion of thrips infected with virus was reduced threefold when insects were preexposed to the G(N)-S protein as compared to no exposure to protein, and nontransmitters were not infected with virus. These results demonstrate that thrips transmission of a tospovirus can be reduced by exogenous viral glycoprotein.

  5. Seasonal dynamics of thrips (Thrips tabaci) (Thysanoptera: Thripidae) transmitters of iris yellow spot virus: a serious viral pathogen of onion bulb and seed crops.

    Science.gov (United States)

    Bag, Sudeep; Rondon, Silvia I; Druffel, Keri L; Riley, David G; Pappu, Hanu R

    2014-02-01

    Thrips-transmitted Iris yellow spot virus (IYSV) is an important economic constraint to the production of bulb and seed onion crops in the United States and many other parts of the world. Because the virus is exclusively spread by thrips, the ability to rapidly detect the virus in thrips vectors would facilitate studies on the role of thrips in virus epidemiology, and thus formulation of better vector management strategies. Using a polyclonal antiserum produced against the recombinant, Escherichia coli-expressed nonstructural protein coded by the small (S) RNA of IYSV, an enzyme linked immunosorbent assay was developed for detecting IYSV in individual as well as groups of adult thrips. The approach enabled estimating the proportion of potential thrips transmitters in a large number of field-collected thrips collected from field-grown onion plants. Availability of a practical and inexpensive test to identify viruliferous thrips would be useful in epidemiological studies to better understand the role of thrips vectors in outbreaks of this economically important virus of onion.

  6. The detection of Tomato spotted wilt virus (TSWV) in individual thrips using real time fluorescent RT-PCR (TaqMan).

    Science.gov (United States)

    Boonham, N; Smith, P; Walsh, K; Tame, J; Morris, J; Spence, N; Bennison, J; Barker, I

    2002-03-01

    Tomato spotted wilt virus (TSWV) is an important virus, economically in the UK, causing damaging disease in ornamental and vegetable crops. The virus is vectored by several species of thrips, most importantly the western flower thrips (Frankliniella occidentalis Pergande [Thysanoptera: Thripidae]). The vector thrips themselves constitute a damaging pest and are difficult to control completely. Monitoring thrips numbers is an important part of the control of virus, but does not give information on how many of the thrips are viruliferous. Monitoring the presence of viruliferous thrips at an early stage of an epidemic may lead to improved disease control, since virus can be spread effectively whilst vector pressure is low and symptoms may take several weeks to appear on some hosts. This paper describes the development of a sensitive and robust, high-throughput method for the detection of TSWV in individual insects based on TaqMan chemistry. The method incorporates a novel RNA specific internal control to increase the reliability of the results. Results are also presented on comparisons of different extraction methods, including insects taken from sticky traps, for high-throughout testing. Implementation of a method such as this for the reliable detection of TSWV in individual thrips would aid the understanding of the progress of TSWV epidemics, and offer an early disease warning system for growers.

  7. Litopenaeus vannamei activating transcription factor 6 alpha gene involvement in ER-stress response and white spot symptom virus infection.

    Science.gov (United States)

    Yuan, Kai; He, Hong-Hui; Zhang, Chao-Zheng; Li, Xiao-Yun; Weng, Shao-Ping; He, Jian-Guo; Chen, Yi-Hong

    2017-11-01

    A previous study found that inositol-requiring enzyme-1-X-box binding protein 1 (IRE1-XBP1) pathway and the protein kinase RNA (PKR)-like ER kinase-eIF2α (PERK-eIF2α) pathway of shrimp play roles in the unfolded protein response (UPR). And they also be proved that was involved in white spot symptom virus (WSSV) infection. Yet the functions of the third branch in shrimp UPR are still unclear. In this study, we showed that upon UPR activation, activating transcription factor 6 alpha (LvATF6α) of Litopenaeus vannamei was cleaved and transferred from the cytoplasm to the nucleus in 293T cells, indicating that the ATF6 pathway in shrimp is also a branch of UPR. Furthermore, LvATF6α could reduce the apoptosis rate of Drosophila Schneider 2 (S2) cells treated with actinomycin, and knock-down expression of LvATF6α increased the apoptosis rate of shrimp hemocytes. In vivo testing revealed that the short from LvATF6α (LvATF6α-s) was obviously increased after UPR activation or WSSV infection, indicating that the ATF6 pathway was activated in L. vannamei gills under such circumstances. Moreover, knock-down expression of LvATF6α could reduce the cumulative mortality and WSSV copy number in WSSV-infected shrimp. Further study revealed that WSSV may profit from shrimp ATF6 pathway activation in two aspects. First, LvATF6α-s significantly upregulated the expression of the WSSV genes (wsv023, wsv045, wsv083, wsv129, wsv222, wsv249, and wsv343). Second, LvATF6α-s inhibited apoptosis by negatively regulating the apoptosis signal-regulating kinase 1 - (c-Jun N-terminal kinase) pathway. All of these evidences suggested that the ATF6 pathway is a member of the L. vannamei UPR, and it is also engaged in WSSV infection. Copyright © 2017 Elsevier Ltd. All rights reserved.

  8. Yeast Surface Display of Two Proteins Previously Shown to Be Protective Against White Spot Syndrome Virus (WSSV in Shrimp.

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    Vorawit Ananphongmanee

    Full Text Available Cell surface display using the yeasts Saccharomyces cerevisiae and Pichia pastoris has been extensively developed for application in bioindustrial processes. Due to the rigid structure of their cell walls, a number of proteins have been successfully displayed on their cell surfaces. It was previously reported that the viral binding protein Rab7 from the giant tiger shrimp Penaeus monodon (PmRab7 and its binding partner envelope protein VP28 of white spot syndrome virus (WSSV could independently protect shrimp against WSSV infection. Thus, we aimed to display these two proteins independently on the cell surfaces of 2 yeast clones with the ultimate goal of using a mixture of the two clones as an orally deliverable, antiviral agent to protect shrimp against WSSV infection. PmRab7 and VP28 were modified by N-terminal tagging to the C-terminal half of S. cerevisiae α-agglutinin. DNA fragments, harboring fused-gene expression cassettes under control of an alcohol oxidase I (AOX1 promoter were constructed and used to transform the yeast cells. Immunofluorescence microscopy with antibodies specific to both proteins demonstrated that mutated PmRab7 (mPmRab7 and partial VP28 (pVP28 were localized on the cell surfaces of the respective clones, and fluorescence intensity for each was significantly higher than that of control cells by flow cytometry. Enzyme-linked immunosorbant assay (ELISA using cells displaying mPmRab7 or pVP28 revealed that the binding of specific antibodies for each was dose-dependent, and could be saturated. In addition, the binding of mPmRab7-expressing cells with free VP28, and vice versa was dose dependent. Binding between the two surface-expressed proteins was confirmed by an assay showing agglutination between cells expressing complementary mPmRab7 and pVP28. In summary, our genetically engineered P. pastoris can display biologically active mPmRab7 and pVP28 and is now ready for evaluation of efficacy in protecting shrimp against

  9. Tipificação de isolados deCurtobacterium flaccumfaciens pv. flaccumfaciens por bacteriocinas

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    Maringoni Antonio Carlos

    2002-01-01

    Full Text Available Este trabalho objetivou avaliar a produção e a sensibilidade à bacteriocinas, de 17 isolados de Curtobacterium flaccumfaciens pv. flaccumfaciens. Doze isolados foram procedentes do Estado de São Paulo e cinco de coleções internacionais. Apenas nove isolados de C. flaccumfaciens pv. flaccumfaciens foram bacteriocinogênicos, e os 17 isolados avaliados foram separados em 12 grupos, conforme a sensibilidade às bacteriocinas produzidas pelos isolados bacteriocinogênicos.

  10. Prevention of White Spot Syndrome Virus Infection on Penaeus monodon by Immersion in CEPM Extract of Avicennia sp. and Sonneratia sp.

    Directory of Open Access Journals (Sweden)

    D. Wahjuningrum

    2007-01-01

    Full Text Available The quality and survival rate are still being the problem that hampers the productivity of black tiger shrimp, Penaeus monodon culture.  Impaired quality of larval shrimp and environmental conditions can confer shrimp be infected by diseases, including viruses such as white spot syndrome virus (WSSV.  Prevention of disease infection using chemicals can offer negative impacts on water, pathogen resistance and consumers.  This study was conducted to examine the efficacy of an alternative prevention compound as liquid mangrove extract (CEPM from Avicennia sp. and Sonneratia sp. By immersion in different dose of CEPM, i.e. 250, 500, 750 and 1000 ppm, the patogenicity of WSSV was found to be different.  Patogenicity of WSSV decreased after treatment by CEPM, hence this could be used to induce shrimp immunity.  Optimum dose of CEPM was 250 ppm, which could increased survival rate of shrimp after challenging by WSSV, up to 98.4% shrimp survived. Keywrods: WSSV, black tiger shrimp, extract, Avicennia sp., Sonneratia sp.   ABSTRAK Kualitas dan kelangsungan hidup merupakan masalah yang masih membatasi produktivitas budidaya udang windu Penaeus monodon. Kondisi udang dan kualitas lingkungan yang kurang baik dapat menyebabkan udang terserang penyakit, termasuk yang disebabkan oleh virus termasuk white spot syndrome virus (WSSV. Upaya pengendaliannya menggunakan bahan kimia secara berlebih dapat menimbulkan dampak negatif bagi lingkungan perairan, kesehatan konsumen dan menimbulkan resistensi patogen. Penelitian ini dilakukan untuk mengetahui efektivitas bahan alternatif berupa cairan ekstrak pohon mangrove (CEPM dari jenis Avicennia sp. dan Sonneratia sp. sebagai upaya pencegahan. Dengan perendaman beberapa konsentrasi yang berbeda (250, 500, 750 dan 1000 ppm penggunaan cairan ekstrak pohon mangrove (CEPM Avicennia sp. dan Sonneratia sp., memberikan pengaruh yang berbeda terhadap patogenitas WSSV dan udang uji pada setiap perlakuan. Tingkat

  11. Second generation peanut genotypes resistant to thrips-transmitted tomato spotted wilt virus exhibit tolerance rather than true resistance and differentially affect thrips fitness.

    Science.gov (United States)

    Shrestha, Anita; Srinivasan, Rajagopalbabu; Sundaraj, Sivamani; Culbreath, Albert K; Riley, David G

    2013-04-01

    Spotted wilt disease caused by Tomato spotted wilt virus (TSWV) (family Bunyaviridae; genus Tospovirus) is a major constraint to peanut (Arachis hypogaea L.) production in the southeastern United States. Reducing yield losses to TSWV has heavily relied on planting genotypes that reduce the incidence of spotted wilt disease. However, mechanisms conferring resistance to TSWV have not been identified in these genotypes. Furthermore, no information is available on how these genotypes influence thrips fitness. In this study, we investigated the effects of newly released peanut genotypes (Georganic, GA-06G, Tifguard, and NC94022) with field resistance to TSWV and a susceptible genotype (Georgia Green) on tobacco thrips, Frankliniella fusca (Hinds), fitness, and TSWV incidence. Thrips-mediated transmission resulted in TSWV infection in both TSWV-resistant and susceptible genotypes and they exhibited typical TSWV symptoms. However, some resistant genotypes had reduced viral loads (fewer TSWV N-gene copies) than the susceptible genotype. F. fusca larvae acquired TSWV from resistant and susceptible genotypes indicating that resistant genotypes also can serve as inoculum sources. Unlike resistant genotypes in other crops that produce local lesions (hypersensitive reaction) upon TSWV infection, widespread symptom development was noticed in peanut genotypes. Results indicated that the observed field resistance in peanut genotypes could be because of tolerance. Further, fitness studies revealed some, but not substantial, differences in thrips adult emergence rates and developmental time between resistant and susceptible genotypes. Thrips head capsule length and width were not different when reared on different genotypes.

  12. Grafting on a Non-Transgenic Tolerant Tomato Variety Confers Resistance to the Infection of a Sw5-Breaking Strain of Tomato spotted wilt virus via RNA Silencing.

    Science.gov (United States)

    Spanò, Roberta; Mascia, Tiziana; Kormelink, Richard; Gallitelli, Donato

    2015-01-01

    RNA silencing controls endogenous gene expression and drives defensive reactions against invasive nucleic acids like viruses. In plants, it has been demonstrated that RNA silencing can be transmitted through grafting between scions and silenced rootstocks to attenuate virus and viroid accumulation in the scions. This has been obtained mostly using transgenic plants, which may be a drawback in current agriculture. In the present study, we examined the dynamics of infection of a resistance-breaking strain of Tomato spotted wilt virus (RB-TSWV) through the graft between an old Apulian (southern Italy) tomato variety, denoted Sl-Ma, used as a rootstock and commercial tomato varieties used as scions. In tests with non-grafted plants, Sl-Ma showed resistance to the RB-TSWV infection as viral RNA accumulated at low levels and plants recovered from disease symptoms by 21 days post inoculation. The resistance trait was transmitted to the otherwise highly susceptible tomato genotypes grafted onto Sl-Ma. The results from the analysis of small RNAs hallmark genes involved in RNA silencing and virus-induced gene silencing suggest that RNA silencing is involved in the resistance showed by Sl-Ma against RB-TSWV and in scions grafted on this rootstock. The results from self-grafted susceptible tomato varieties suggest also that RNA silencing is enhanced by the graft itself. We can foresee interesting practical implications of the approach described in this paper.

  13. Grafting on a Non-Transgenic Tolerant Tomato Variety Confers Resistance to the Infection of a Sw5-Breaking Strain of Tomato spotted wilt virus via RNA Silencing.

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    Roberta Spanò

    Full Text Available RNA silencing controls endogenous gene expression and drives defensive reactions against invasive nucleic acids like viruses. In plants, it has been demonstrated that RNA silencing can be transmitted through grafting between scions and silenced rootstocks to attenuate virus and viroid accumulation in the scions. This has been obtained mostly using transgenic plants, which may be a drawback in current agriculture. In the present study, we examined the dynamics of infection of a resistance-breaking strain of Tomato spotted wilt virus (RB-TSWV through the graft between an old Apulian (southern Italy tomato variety, denoted Sl-Ma, used as a rootstock and commercial tomato varieties used as scions. In tests with non-grafted plants, Sl-Ma showed resistance to the RB-TSWV infection as viral RNA accumulated at low levels and plants recovered from disease symptoms by 21 days post inoculation. The resistance trait was transmitted to the otherwise highly susceptible tomato genotypes grafted onto Sl-Ma. The results from the analysis of small RNAs hallmark genes involved in RNA silencing and virus-induced gene silencing suggest that RNA silencing is involved in the resistance showed by Sl-Ma against RB-TSWV and in scions grafted on this rootstock. The results from self-grafted susceptible tomato varieties suggest also that RNA silencing is enhanced by the graft itself. We can foresee interesting practical implications of the approach described in this paper.

  14. Role of Oxalis corniculata L. as plant virus reservoir with special regard to Tomato Spotted Wilt Virus (TSWV-RB strain occurrence in rock-wool cultivation in Hungary

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    Hodi, Anna Maria

    2014-02-01

    Full Text Available The creeping wood sorrel (Oxalis corniculata L. in Central Europe occurs mainly in anthropogenic areas, where it grows in parks and landfill sites or in protected cultivation sites like greenhouses and nurseries. In Hungary O. corniculata has been spreading since the 19th century. In glass- and greenhouses it is the only weed species that was able to settle and flourish on hydroponic rock-wool and coconut fibre growing media. Among virus pathogens of forced tomato (LYPES and paprika crops (CPSAN in Hungary the Tomato Spotted Wilt Virus (TSWV has become an important one, severely affecting the plants and on occasions causing 100% yield losses. The spread of the virus was largely assisted by its effective vector the Western Flower Thrips (Frankliniella occidentalis Pergande. Initially the control of the disease was based on the control of the thrips, later, virus-resistant varieties were introduced. This however proved insufficient to fight the disease as it was discovered, quite recently, that in certain white pepper (CPSAN varieties the virus was able to break up the resistance. According to data of the year 2012, those varieties that were considered resistant, showed up to 50% virus infection. Considering the wide spread of O. corniculata and increasing occurrence of the resistance-breaking TSWV isolates, surveys in greenhouses were conducted to examine whether the weed could serve as reservoir for TSWV. Samples were collected of the virus infected crop plants and O. corniculata growing in the same coconut fibre cubes. The samples were examined using test-plants, serological and RT-PCR methods. In result it was found that the symptoms were indeed caused by resistance-breaking TSWV isolates. However, no virus was found in the suspected O. corniculata samples. Therefore it was concluded that in this particular case the O. corniculata cannot be considered reservoir for the virus, in the hydroponic culture.

  15. Incidência de vírus em videiras no Nordeste brasileiro e caracterização molecular parcial de isolados virais locais

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    Aricléia de Moraes Catarino

    2015-03-01

    Full Text Available Os objetivos deste trabalho foram identificar as espécies virais presentes em vinhedos comerciais de duas regiões do Nordeste do Brasil e realizar a caracterização molecular parcial de isolados de três espécies virais. A diagnose foi realizada por meio de RT-PCR em tempo real para a detecção de Grapevine rupestris stem pitting-associated virus (GRSPaV, Grapevine virus A (GVA, Grapevine virus B (GVB, Grapevine leafroll-associated virus 2, 3 e 4 (GLRaV-2, -3 e -4, Grapevine fleck virus (GFkV, Grapevine rupestris vein feathering virus (GRVFV e Grapevine fanleaf virus (GFLV. Exceto para GFLV, os vírus avaliados estão amplamente disseminados nas áreas amostradas, frequentemente em altas incidências e em infecções múltiplas, de até 98% e 76,4%, na Zona da Mata e no Vale do São Francisco, respectivamente. Isolados locais de GVA, GVB e GLRaV-3 foram parcialmente caracterizados com base na sequência completa de nucleotídeos do gene da proteína capsidial e apresentaram alta porcentagem de identidade de nucleotídeos com outros isolados brasileiros: 91,2% (GVA, 99,8% (GVB e 99,7% (GLRaV-3

  16. Lateral flow assay for rapid detection of white spot syndrome virus (WSSV) using a phage-displayed peptide as bio-recognition probe.

    Science.gov (United States)

    Kulabhusan, Prabir Kumar; Rajwade, Jyutika M; Sahul Hameed, A S; Paknikar, Kishore M

    2017-06-01

    White spot disease caused by the white spot syndrome virus (WSSV) has a major socio-economic impact on shrimp farming in India. It has been realized that a field-usable diagnostic capable of rapid detection of WSSV can prevent huge economic losses in disease outbreaks. In this work, we explored the possibility of using a peptide as bio-recognition probe in a field-usable device for the detection of WSSV from infected shrimps and prawns. A commercially available random phage-display library was screened against rVP28 (a major structural protein of WSSV, expressed as a recombinant protein in Escherichia coli). A bacteriophage clone VP28-4L was obtained, and its binding to purified rVP28 protein as well as WSSV from infected shrimp Litopaeneus vannamei tissue was confirmed by ELISA and western blot. The apparent equilibrium dissociation constant (K d ,app) was calculated to be 810 nM. VP28-4L did not show cross-reactivity with any other shrimp viruses. A 12-mer peptide (pep28, with the sequence 'TFQAFDLSPFPS') displayed on the VP28-4L was synthesized, and its diagnostic potential was evaluated in a lateral flow assay (LFA). Visual detection of WSSV could be achieved using biotinylated-pep28 and streptavidin-conjugated gold nanoparticles. In LFA, 12.5 μg/mL of the virus could be detected from L. vannamei gill tissue homogenate within 20 min. Pep28 thus becomes an attractive candidate in bio-recognition of WSSV in field-usable diagnostic platforms benefitting the aquaculture sector.

  17. Sequence characterization, molecular phylogeny reconstruction and recombination analysis of the large RNA of Tomato spotted wilt virus (Tospovirus: Bunyaviridae) from the United States.

    Science.gov (United States)

    Ramesh, Shunmugiah V; Pappu, Hanu R

    2016-04-01

    Tomato spotted wilt virus (TSWV; Tospovirus: Bunyaviridae) has been an economically important virus in the USA for over 30 years. However the complete sequence of only one TSWV isolate PA01 characterized from pepper in Pennsylvania is available. The large (L) RNA of a TSWV WA-USA isolate was cloned and sequenced. It consisted of 8914 nucleotides (nt) encoding a single open reading frame of 8640 nts in the viral-complementary sense. The ORF potentially codes for RNA-dependent RNA polymerase (RdRp) of 330.9 kDa. Two untranslated regions of 241 and 33 nucleotides were present at the 5' and 3' termini, respectively that shared conserved tospoviral sequences. Phylogenetic analysis using nucleotide sequences of the complete L RNA showed that TSWV WA-USA isolate clustered with the American and Asian TSWV isolates which formed a distinct clade from Euro-Asiatic Tospoviruses. Phylogeny of the amino acid sequence of all tospoviral RdRps used in this study showed that all the known TSWV isolates including the USA isolate described in this study formed a distinct and a close cluster with that of Impateins necrotic spot virus. Multiple sequence alignment revealed conserved motifs in the RdRp of TSWV. Recombination analysis identified two recombinants including the TSWV WA-USA isolate. Among them, three recombination events were detected in the conserved motifs of the RdRp. Sequence analysis and phylogenetic analysis of the L RNA showed distinct clustering with selected TSWV isolates reported from elsewhere. Conserved motifs in the core polymerase region of the RdRp and recombination events were identified.

  18. SUMO-conjugating enzyme E2 UBC9 mediates viral immediate-early protein SUMOylation in crayfish to facilitate reproduction of white spot syndrome virus.

    Science.gov (United States)

    Chen, An-Jing; Gao, Lu; Wang, Xian-Wei; Zhao, Xiao-Fan; Wang, Jin-Xing

    2013-01-01

    Successful viruses have evolved superior strategies to escape host defenses or exploit host biological pathways. Most of the viral immediate-early (ie) genes are essential for viral infection and depend solely on host proteins; however, the molecular mechanisms are poorly understood. In this study, we focused on the modification of viral IE proteins by the crayfish small ubiquitin-related modifier (SUMO) and investigated the role of SUMOylation during the viral life cycle. SUMO and SUMO ubiquitin-conjugating enzyme 9 (UBC9) involved in SUMOylation were identified in red swamp crayfish (Procambarus clarkii). Both SUMO and UBC9 were upregulated in crayfish challenged with white spot syndrome virus (WSSV). Replication of WSSV genes increased in crayfish injected with recombinant SUMO or UBC9, but injection of mutant SUMO or UBC9 protein had no effect. Subsequently, we analyzed the mechanism by which crayfish SUMOylation facilitates WSSV replication. Crayfish UBC9 bound to all three WSSV IE proteins tested, and one of these IE proteins (WSV051) was covalently modified by SUMO in vitro. The expression of viral ie genes was affected and that of late genes was significantly inhibited in UBC9-silenced or SUMO-silenced crayfish, and the inhibition effect was rescued by injection of recombinant SUMO or UBC9. The results of this study demonstrate that viral IE proteins can be modified by crayfish SUMOylation, prompt the expression of viral genes, and ultimately benefit WSSV replication. Understanding of the mechanisms by which viruses exploit host components will greatly improve our knowledge of the virus-host "arms race" and contribute to the development of novel methods against virulent viruses.

  19. White-spot syndrome virus (WSSV) introduction into the Gulf of Mexico and Texas freshwater systems through imported, frozen bait-shrimp.

    Science.gov (United States)

    Hasson, K W; Fan, Y; Reisinger, T; Venuti, J; Varner, P W

    2006-07-25

    We analysed 20 boxes of, frozen imported bait-shrimp (China: Parapenaeopsis sp. and Metapenaeopsis sp.) and 8 boxes of native, frozen bait-shrimp (Gulf of Mexico: Litopenaeus setiferus and Farfantepenaeus duorarum) by RT-PCR or PCR for Taura syndrome virus (TSV), yellowhead virus/gill-associated virus (YHV/GAV), white-spot syndrome virus (WSSV) and infectious hypodermal and hematopoietic necrosis virus (IHHNV). All 28 boxes of shrimp were negative for TSV, YHV/GAV and IHHNV; 2 boxes of imported bait-shrimp were WSSV-positive by 3 different PCR assays. Intramuscular injection of replicate groups of SPF (specific pathogen-free) L. vannamei juveniles with 2 different tissue homogenates prepared from the 2 WSSV-positive bait boxes resulted in 100% mortality of the test shrimp within 48 to 72 h post-injection. No mortality occurred among injected negative control groups. Histological and in situ hybridization analyses of 20 moribund treatment-shrimp demonstrated severe WSSV infections in each sample. Oral exposure of SPF L. vannamei postlarvae, PL (PL 25 to 30 stage; approximately 0.02 g) to minced tissue prepared from the 2 WSSV-positive bait-lots did not induce infection, possibly because of an insufficient infectious dose and/or viral inactivation resulting from multiple freeze-thaw cycles of the bait-shrimp during PCR testing. Use of an electric drill and collection of drill-tailings (tissue from approximately 20 to 30 shrimp) from frozen blocks of shrimp was successfully employed as an alternate tissue-sampling method without thawing. Our findings indicate that imported WSSV-infected bait shrimp, originating from China, are being sold in Texas for the purpose of sport fishing and represent a potential threat to freshwater and marine crustacean fisheries, as well as to coastal US shrimp farms.

  20. Age Spots

    Science.gov (United States)

    ... Hands Age Spots Aging Skin Birthmarks Burn Scars Cellulite Crow's Feet Droopy Eyelids Excess Fat Excessive Sweating ... Hands Age Spots Aging Skin Birthmarks Burn Scars Cellulite Crow's Feet Droopy Eyelids Excess Fat Excessive Sweating ...

  1. Host MicroRNA-217 Promotes White Spot Syndrome Virus Infection by Targeting Tube in the Chinese Mitten Crab (Eriocheir sinensis

    Directory of Open Access Journals (Sweden)

    Ying Huang

    2017-05-01

    Full Text Available MicroRNAs (miRNAs, a group of small molecule non-encoding RNAs, are key post-transcriptional regulators of gene expression that are implicated in many biological processes. In the current study, miR-217 from Eriocheir sinensis was selected for studying its roles during host–virus interaction. Overexpression or silencing of miR-217 led to considerable effects on white spot syndrome virus (WSSV replication, implying that miR-217 played a positive role in WSSV infection. In insect High Five cells, miR-217 significantly inhibited Tube gene expression by binding to the 3′-untranslated region of the Tube. Overexpression of miR-217 in crab led to downregulation of tube expression. Knockdown of Tube in vivo led to significant enhancement of WSSV infection and inhibited the expression of five antimicrobial peptide (AMP genes (Anti-lipopolysaccharide factor ALF1, ALF2, ALF3; Crustin Crus1, Crus2 in WSSV-challenged crabs. Overexpression of miR-217 also led to downregulation of these AMP genes in WSSV-challenged crabs. Our results showed that host miRNA played positive roles in virus infection by regulation of host tube gene, which is the key component of Toll signaling pathway.

  2. Pereirina: o primeiro alcaloide isolado no Brasil?

    Directory of Open Access Journals (Sweden)

    Márcia R. Almeida

    Full Text Available O artigo descreve a história da pereirina, um alcaloide extraído das cascas da Geissospermum vellosii Allemão, uma espécie de planta brasileira da família Apocynaceae, conhecida popularmente como Pau-pereira. A classificação botânica desta espécie e o emprego da pereirina na medicina popular foram as razões que acirraram a disputa entre os farmacêuticos brasileiros e estrangeiros pela glória de sua descoberta no Brasil do século XIX. Neste estudo constatou-se que a glória desta descoberta é do farmacêutico brasileiro Ezequiel Corrêa dos Santos e desta maneira, a pereirina é o primeiro alcaloide isolado no Brasil. As cascas do Pau-pereira foram, durante dois séculos, um importante remédio no combate a diversas doenças, devido às propriedades terapêuticas dos alcaloides presentes nas cascas da espécie e atualmente pesquisas estão sendo realizadas para comprovação científica de sua eficácia no combate a doenças virais como Aids, herpes e hepatite C.

  3. Host plant resistance against tomato spotted wilt virus in peanut (Arachis hypogaea) and its impact on susceptibility to the virus, virus population genetics, and vector feeding behavior and survival.

    Science.gov (United States)

    Sundaraj, Sivamani; Srinivasan, Rajagopalbabu; Culbreath, Albert K; Riley, David G; Pappu, Hanu R

    2014-02-01

    Tomato spotted wilt virus (TSWV) severely affects peanut production in the southeastern United States. Breeding efforts over the last three decades resulted in the release of numerous peanut genotypes with field resistance to TSWV. The degree of field resistance in these genotypes has steadily increased over time, with recently released genotypes exhibiting a higher degree of field resistance than older genotypes. However, most new genotypes have never been evaluated in the greenhouse or laboratory against TSWV or thrips, and the mechanism of resistance is unknown. In this study, TSWV-resistant and -susceptible genotypes were subjected to TSWV mechanical inoculation. The incidence of TSWV infection was 71.7 to 87.2%. Estimation of TSWV nucleocapsid (N) gene copies did not reveal significant differences between resistant and susceptible genotypes. Parsimony and principal component analyses of N gene nucleotide sequences revealed inconsistent differences between virus isolates collected from resistant and susceptible genotypes and between old (collected in 1998) and new (2010) isolates. Amino acid sequence analyses indicated consistent differences between old and new isolates. In addition, we found evidence for overabundance of nonsynonymous substitutions. However, there was no evidence for positive selection. Purifying selection, population expansion, and differentiation seem to have influenced the TSWV populations temporally rather than positive selection induced by host resistance. Choice and no-choice tests indicated that resistant and susceptible genotypes differentially affected thrips feeding and survival. Thrips feeding and survival were suppressed on some resistant genotypes compared with susceptible genotypes. These findings reveal how TSWV resistance in peanut could influence evolution, epidemiology, and management of TSWV.

  4. Genetic and host-associated differentiation within Thrips tabaci Lindeman (Thysanoptera: Thripidae) and its links to Tomato spotted wilt virus-vector competence.

    Science.gov (United States)

    Westmore, G C; Poke, F S; Allen, G R; Wilson, C R

    2013-09-01

    Of eight thelytokous populations of onion thrips (Thrips tabaci) collected from potato (three populations), onion (four) or Chrysanthemum (one) hosts from various regions of Australia, only those from potato were capable of transmitting Tomato spotted wilt virus (TSWV) in controlled transmission experiments. Genetic differentiation of seven of these eight populations, and nine others not tested for TSWV vector competence, was examined by comparison of the DNA sequences of mitochondrial cytochrome oxidase subunit 1 (COI) gene. All Australian populations of T. tabaci grouped within the European 'L2' clade of Brunner et al. (2004). Within this clade the seven populations from potato, the three from onion, and the four from other hosts (Chrysanthemum, Impatiens, lucerne, blackberry nightshade) clustered as three distinct sub-groupings characterised by source host. Geographical source of thrips populations had no influence on genetic diversity. These results link genetic differentiation of thelytokous T. tabaci to source host and to TSWV vector capacity for the first time.

  5. Molecular cloning and recombinant expression of the VP28 carboxyl-terminal hydrophilic region from a brazilian white spot syndrome virus isolate

    Directory of Open Access Journals (Sweden)

    Patricia Braunig

    2011-04-01

    Full Text Available In the present study, a fragment of the VP28 coding sequence from a Brazilian WSSV isolate (BrVP28 was cloned, sequenced and expressed in E. coli BL21(DE3 pLysS strain in order to produce the VP28 carboxyl-terminal hydrophilic region. The expression resulted in a protein of about 21 kDa, which was purified under denaturing conditions, resulting in a final highly purified BrVP28 preparation. The recombinant protein obtained can be used in several biotechnology applications, such as the production of monoclonal antibodies which could be used in the development of diagnostic tools as well as in the studies on the characterization of white spot syndrome virus (WSSV isolated in Brazil.

  6. Mutational analysis of two highly conserved motifs in the silencing suppressor encoded by tomato spotted wilt virus (genus Tospovirus, family Bunyaviridae).

    Science.gov (United States)

    Zhai, Ying; Bag, Sudeep; Mitter, Neena; Turina, Massimo; Pappu, Hanu R

    2014-06-01

    Tospoviruses cause serious economic losses to a wide range of field and horticultural crops on a global scale. The NSs gene encoded by tospoviruses acts as a suppressor of host plant defense. We identified amino acid motifs that are conserved in all of the NSs proteins of tospoviruses for which the sequence is known. Using tomato spotted wilt virus (TSWV) as a model, the role of these motifs in suppressor activity of NSs was investigated. Using site-directed point mutations in two conserved motifs, glycine, lysine and valine/threonine (GKV/T) at positions 181-183 and tyrosine and leucine (YL) at positions 412-413, and an assay to measure the reversal of gene silencing in Nicotiana benthamiana line 16c, we show that substitutions (K182 to A, and L413 to A) in these motifs abolished suppressor activity of the NSs protein, indicating that these two motifs are essential for the RNAi suppressor function of tospoviruses.

  7. First report of tomato chlorotic spot virus in non-solanaceous weeds erect spiderling (Boerhavia erecta) and asian spiderflower (Cleome viscosa), and sweet chili pepper (Capsicum chinense) in Puerto Rico

    Science.gov (United States)

    Tomato chlorotic spot virus (TCSV) has recently been detected in tomato, bell pepper, jimsonweed and lettuce in Puerto Rico. Observations of weeds and additional crops in 2015 and 2016 revealed TCSV-like symptoms. Testing of these symptomatic plants identified three new hosts of TCSV in Puerto Ric...

  8. Application of Spirulina platensis for prevention of white spot syndrome virus in post larvae and juvenile black tiger shrimp (Penaeus monodon

    Directory of Open Access Journals (Sweden)

    Hemtanon, P.

    2005-02-01

    Full Text Available In this study, the extract of Spirulina platensis were examined in vitro to inhibit white spot syndrome virus (WSSV and application of dry S. platensis in diet for prevention of white spot syndrome (WSS in post larvae and juvenile black tiger shrimp (Penaeus monodon. The results showed that the lowest concentration of the extract for inhibiting WSSV was 0.01 mg/ml, while the optimum concentration was found to be 0.1 mg/ ml in which the mortality rate of the shrimp was 4 percents and infection was not detected from survivalshrimp by the immunohistochemistry method.Furthermore, The results showed that the survival rate of the post larvae fed on steamed egg containing dry S. platensis 5 g/kg of diet was higher than that of the control (p<0.05 when challenged with WSSV and no WSSV infected shrimp examine by polymerase chain reaction (PCR assay. In the case of juvenile shrimp, the survival rate of shrimp fed pellets containing dry S. platensis 10 g/kg of diet was higher than that of the control group (p<0.05 after challenging with WSSV. Moreover percent of WSSV infection in the survival shrimp using the immunohistochemistry method was lower than that of the control group.

  9. Evaluation of peanut genotypes for resistance to Tomato spotted wilt virus by mechanical and thrips inoculation Avaliação de genótipos de amendoim em relação à resistência ao Tomato spotted wilt virus por meio de inoculação mecânica e por tripes

    OpenAIRE

    Luciana Cordeiro do Nascimento; Viboon Pensuk; Nivânia Pereira da Costa; Francisco Miguel de Assis Filho; Gilvan Pio-Ribeiro; Carl Michael Deom; John Sherwood

    2006-01-01

    The objective of this work was to evaluate the reactions of three peanut breeding lines (IC-10, IC-34, and ICGV 86388) to Tomato spotted wilt virus (TSWV) by mechanical and thrips inoculation, under greenhouse conditions, and compare them to the reactions of cultivars SunOleic, Georgia Green, and the breeding line C11-2-39. TSWV infection by mechanical inoculation was visually assessed using an index ranging from 0 (no symptoms) to 4 (apical death). Enzyme-linked immunosorbent assay was used ...

  10. The Tomato spotted wilt virus genome is processed differentially in its plant host Arachis hypogaea and its thrips vector Frankliniella fusca

    Directory of Open Access Journals (Sweden)

    Stephen John Fletcher

    2016-09-01

    Full Text Available Thrips-transmitted tospoviruses are economically important viruses affecting a wide range of field and horticultural crops worldwide. Tomato spotted wilt virus is the type member of the Tospovirus genus with a broad host range of more than 900 plant species. Interactions between these viruses and their plant hosts and insect vectors via RNA interference pathways are likely a key determinant of pathogenicity. The current investigation, for the first time, compares biogenesis of small RNAs between the plant host and insect vector in the presence or absence of TSWV. Unique viral small interfering RNA (vsiRNA profiles are evident for Arachis hypogaea (peanut and Frankliniella fusca (thrips vector following infection with TSWV. Differences between vsiRNA profiles for these plant and insect species, such as the relative abundance of 21 nt and 22 nt vsiRNAs and locations of alignment hotspots, reflect the diverse siRNA biosynthesis pathways of their respective kingdoms. The presence of unique vsiRNAs in F. fusca samples indicates that vsiRNA generation takes place within the thrips, and not solely through uptake via feeding on vsiRNAs produced in infected A. hypogaea. The study also shows key vsiRNA profile differences for TSWV among plant families, which are evident in the case of A. hypogaea, a legume, and members of Solanaceae (S. lycopersicum and N. benthamiana. Distinctively, overall small RNA biogenesis in A. hypogaea is markedly affected with an absence of the 24 nt small RNAs in TSWV-infected plants, possibly leading to wide-spread molecular and phenotypic perturbations specific to this species. These findings add significant information on the host-virus-vector interaction in terms of RNAi pathways and may lead to better crop and vector specific control strategies.

  11. The Tomato Spotted Wilt Virus Genome Is Processed Differentially in its Plant Host Arachis hypogaea and its Thrips Vector Frankliniella fusca.

    Science.gov (United States)

    Fletcher, Stephen J; Shrestha, Anita; Peters, Jonathan R; Carroll, Bernard J; Srinivasan, Rajagopalbabu; Pappu, Hanu R; Mitter, Neena

    2016-01-01

    Thrips-transmitted tospoviruses are economically important viruses affecting a wide range of field and horticultural crops worldwide. Tomato spotted wilt virus (TSWV) is the type member of the Tospovirus genus with a broad host range of more than 900 plant species. Interactions between these viruses and their plant hosts and insect vectors via RNAi pathways are likely a key determinant of pathogenicity. The current investigation, for the first time, compares biogenesis of small RNAs between the plant host and insect vector in the presence or absence of TSWV. Unique viral small interfering RNA (vsiRNA) profiles are evident for Arachis hypogaea (peanut) and Frankliniella fusca (thrips vector) following infection with TSWV. Differences between vsiRNA profiles for these plant and insect species, such as the relative abundance of 21 and 22 nt vsiRNAs and locations of alignment hotspots, reflect the diverse siRNA biosynthesis pathways of their respective kingdoms. The presence of unique vsiRNAs in F. fusca samples indicates that vsiRNA generation takes place within the thrips, and not solely through uptake via feeding on vsiRNAs produced in infected A. hypogaea. The study also shows key vsiRNA profile differences for TSWV among plant families, which are evident in the case of A. hypogaea, a legume, and members of Solanaceae (S. lycopersicum and Nicotiana benthamiana). Distinctively, overall small RNA (sRNA) biogenesis in A. hypogaea is markedly affected with an absence of the 24 nt sRNAs in TSWV-infected plants, possibly leading to wide-spread molecular and phenotypic perturbations specific to this species. These findings add significant information on the host-virus-vector interaction in terms of RNAi pathways and may lead to better crop and vector specific control strategies.

  12. Genotype-specific real-time PCR combined with high-resolution melting analysis for rapid identification of red-spotted grouper nervous necrosis virus.

    Science.gov (United States)

    Toubanaki, Dimitra K; Karagouni, Evdokia

    2017-08-01

    A real-time genotype-specific polymerase chain reaction (PCR) assay combined with high-resolution melting (HRM) analysis was developed to assess the most common genotypes of nervous necrosis viruses or nodaviruses. Nodaviruses are the causal agents of viral nervous necrosis infections, which have been wreaking havoc in the aquaculture industry worldwide, with fish mortality up to 100%. The four different genotypes of nodaviruses correlate with differences in viral pathogenicity. Therefore, rational development of effective vaccines and diagnostics requires analysis of genetic variation among viruses. The aim of the present study was to develop a real-time tetra-primer genotype-specific PCR assay for genotype identification. Four primers were utilized for simultaneous amplification of nodavirus genotype-specific products in a single closed-tube PCR after a reverse-transcription reaction using RNA isolated from fish samples. For high-throughput sample analysis, SYBR Green-based real-time PCR was used in combination with HRM analysis. The assay was evaluated in terms of specificity and sensitivity. The analysis resulted in melting curves that were indicative of each genotype. The detection limit when using reference plasmids was 100 ag/µL for both genotypes, while the sensitivity of the assays when testing a complex mixture was 10 fg/µL for red-spotted grouper nervous necrosis virus (RGNNV) and 100 fg/µL for striped jack nervous necrosis virus (SJNNV). To test the capability of this method under real-world conditions, 58 samples were examined. All samples belonged to the RGNNV genotype, which was fully validated. The results were in full agreement with genotyping by reference methods. The proposed methodology provides a rapid, sensitive, specific, robust and automatable assay for nodavirus genotyping, making it a useful tool for diagnosis and screening for epidemiological studies.

  13. Validation of a commercial insulated isothermal PCR-based POCKIT test for rapid and easy detection of white spot syndrome virus infection in Litopenaeus vannamei.

    Directory of Open Access Journals (Sweden)

    Yun-Long Tsai

    Full Text Available Timely pond-side detection of white spot syndrome virus (WSSV plays a critical role in the implementation of bio-security measures to help minimize economic losses caused by white spot syndrome disease, an important threat to shrimp aquaculture industry worldwide. A portable device, namely POCKIT™, became available recently to complete fluorescent probe-based insulated isothermal PCR (iiPCR, and automatic data detection and interpretation within one hour. Taking advantage of this platform, the IQ Plus™ WSSV Kit with POCKIT system was established to allow simple and easy WSSV detection for on-site users. The assay was first evaluated for its analytical sensitivity and specificity performance. The 95% limit of detection (LOD of the assay was 17 copies of WSSV genomic DNA per reaction (95% confidence interval [CI], 13 to 24 copies per reaction. The established assay has detection sensitivity similar to that of OIE-registered IQ2000™ WSSV Detection and Protection System with serial dilutions of WSSV-positive Litopenaeus vannamei DNA. No cross-reaction signals were generated from infectious hypodermal and haematopoietic necrosis virus (IHHNV, monodon baculovirus (MBV, and hepatopancreatic parvovirus (HPV positive samples. Accuracy analysis using 700 L. vannamei of known WSSV infection status shows that the established assayhassensitivity93.5% (95% CI: 90.61-95.56% and specificity 97% (95% CI: 94.31-98.50%. Furthermore, no discrepancy was found between the two assays when 100 random L. vannamei samples were tested in parallel. Finally, excellent correlation was observed among test results of three batches of reagents with 64 samples analyzed in three different laboratories. Working in a portable device, IQ Plus™ WSSV Kit with POCKIT system allows reliable, sensitive and specific on-site detection of WSSV in L. vannamei.

  14. Detection of infections with hepatitis B virus, hepatitis C virus, and human immunodeficiency virus by analyses of dried blood spots - performance characteristics of the ARCHITECT system and two commercial assays for nucleic acid amplification

    Science.gov (United States)

    2013-01-01

    Background Nowadays, dried blood spots (DBS) are primarily used to obtain diagnostic access to risk collectives such as intravenous drug users, who are prone to infections with hepatitis B virus (HBV), hepatitis C virus (HCV), and human immunodeficiency virus (HIV). Before DBS analyses can be used in this diagnostic context, however, a comprehensive evaluation of its performance characteristics must be conducted. To the best of our knowledge, the current study presents for the first time such essential data for the Abbott ARCHITECT system, which is currently the worldwide leading platform in this field of infection diagnostics. Methods The investigation comprised 1,762 paired serum/DBS samples and a total of 3,524 determinations with the Abbott ARCHITECT HBsAg, anti-HBc, anti-HBs, anti-HCV and HIV-1-p24-antigen/anti-HIV 1/2 assays as well as with the artus HBV LC PCR and VERSANT HCV RNA qualitative (TMA) tests. Results In the context of DBS testing, a specificity of 100% was recorded for the seven serological and molecular biological assays. The analytical sensitivity of HBsAg, anti-HBc, anti-HBs, anti-HCV, HIV-1-p24-antigen/anti-HIV 1/2, HBV DNA, and HCV RNA detections in DBS eluates was 98.6%, 97.1%, 97.5%, 97.8%, 100%, 93%, and 100%, respectively. Discussion/conclusions The results obtained indicate that it is today possible to reliably detect HBsAg, anti-HBc, anti-HBs, anti-HCV and HIV-1-p24 antigen/anti-HIV 1/2 with state-of-the-art analytical systems such as the Abbott ARCHITECT in DBS eluates even when a comparatively high elution volume of 1,000 μl is used. They also provide evidence for the inherent analytical limits of DBS testing, which primarily concern the anti-HBc/anti-HBs system for individuals with HIV infections and nucleic acid tests with relatively low analytical sensitivity. PMID:23497102

  15. Detection of infections with hepatitis B virus, hepatitis C virus, and human immunodeficiency virus by analyses of dried blood spots--performance characteristics of the ARCHITECT system and two commercial assays for nucleic acid amplification.

    Science.gov (United States)

    Ross, R Stefan; Stambouli, Oumaima; Grüner, Nico; Marcus, Ulrich; Cai, Wei; Zhang, Weidong; Zimmermann, Ruth; Roggendorf, Michael

    2013-03-05

    Nowadays, dried blood spots (DBS) are primarily used to obtain diagnostic access to risk collectives such as intravenous drug users, who are prone to infections with hepatitis B virus (HBV), hepatitis C virus (HCV), and human immunodeficiency virus (HIV). Before DBS analyses can be used in this diagnostic context, however, a comprehensive evaluation of its performance characteristics must be conducted. To the best of our knowledge, the current study presents for the first time such essential data for the Abbott ARCHITECT system, which is currently the worldwide leading platform in this field of infection diagnostics. The investigation comprised 1,762 paired serum/DBS samples and a total of 3,524 determinations with the Abbott ARCHITECT HBsAg, anti-HBc, anti-HBs, anti-HCV and HIV-1-p24-antigen/anti-HIV 1/2 assays as well as with the artus HBV LC PCR and VERSANT HCV RNA qualitative (TMA) tests. In the context of DBS testing, a specificity of 100% was recorded for the seven serological and molecular biological assays. The analytical sensitivity of HBsAg, anti-HBc, anti-HBs, anti-HCV, HIV-1-p24-antigen/anti-HIV 1/2, HBV DNA, and HCV RNA detections in DBS eluates was 98.6%, 97.1%, 97.5%, 97.8%, 100%, 93%, and 100%, respectively. The results obtained indicate that it is today possible to reliably detect HBsAg, anti-HBc, anti-HBs, anti-HCV and HIV-1-p24 antigen/anti-HIV 1/2 with state-of-the-art analytical systems such as the Abbott ARCHITECT in DBS eluates even when a comparatively high elution volume of 1,000 μl is used. They also provide evidence for the inherent analytical limits of DBS testing, which primarily concern the anti-HBc/anti-HBs system for individuals with HIV infections and nucleic acid tests with relatively low analytical sensitivity.

  16. Molecular docking and simulation studies of 3-(1-chloropiperidin-4-yl)-6-fluoro benzisoxazole 2 against VP26 and VP28 proteins of white spot syndrome virus.

    Science.gov (United States)

    Sudharsana, S; Rajashekar Reddy, C B; Dinesh, S; Rajasekhara Reddy, S; Mohanapriya, A; Itami, T; Sudhakaran, R

    2016-10-01

    White spot syndrome virus (WSSV), an aquatic virus infecting shrimps and other crustaceans, is widely distributed in Asian subcontinents including India. The infection has led to a serious economic loss in shrimp farming. The WSSV genome is approximately 300 kb and codes for several proteins mediating the infection. The envelope proteins VP26 and VP28 play a major role in infection process and also in the interaction with the host cells. A comprehensive study on the viral proteins leading to the development of safe and potent antiviral therapeutic is of adverse need. The novel synthesized compound 3-(1-chloropiperidin-4-yl)-6-fluoro benzisoxazole 2 is proved to have potent antiviral activity against WSSV. The compound antiviral activity is validated in freshwater crabs (Paratelphusa hydrodomous). An in silico molecular docking and simulation analysis of the envelope proteins VP26 and VP28 with the ligand 3-(1-chloropiperidin-4-yl)-6-fluoro benzisoxazole 2 are carried out. The docking analysis reveals that the polar amino acids in the pore region of the envelope proteins were involved in the ligand binding. The influence of the ligand binding on the proteins is validated by the molecular dynamics and simulation study. These in silico approaches together demonstrate the ligand's efficiency in preventing the trimers from exhibiting their physiological function. © 2016 John Wiley & Sons Ltd.

  17. The Tomato Spotted Wilt Virus Genome Is Processed Differentially in its Plant Host Arachis hypogaea and its Thrips Vector Frankliniella fusca

    Science.gov (United States)

    Fletcher, Stephen J.; Shrestha, Anita; Peters, Jonathan R.; Carroll, Bernard J.; Srinivasan, Rajagopalbabu; Pappu, Hanu R.; Mitter, Neena

    2016-01-01

    Thrips-transmitted tospoviruses are economically important viruses affecting a wide range of field and horticultural crops worldwide. Tomato spotted wilt virus (TSWV) is the type member of the Tospovirus genus with a broad host range of more than 900 plant species. Interactions between these viruses and their plant hosts and insect vectors via RNAi pathways are likely a key determinant of pathogenicity. The current investigation, for the first time, compares biogenesis of small RNAs between the plant host and insect vector in the presence or absence of TSWV. Unique viral small interfering RNA (vsiRNA) profiles are evident for Arachis hypogaea (peanut) and Frankliniella fusca (thrips vector) following infection with TSWV. Differences between vsiRNA profiles for these plant and insect species, such as the relative abundance of 21 and 22 nt vsiRNAs and locations of alignment hotspots, reflect the diverse siRNA biosynthesis pathways of their respective kingdoms. The presence of unique vsiRNAs in F. fusca samples indicates that vsiRNA generation takes place within the thrips, and not solely through uptake via feeding on vsiRNAs produced in infected A. hypogaea. The study also shows key vsiRNA profile differences for TSWV among plant families, which are evident in the case of A. hypogaea, a legume, and members of Solanaceae (S. lycopersicum and Nicotiana benthamiana). Distinctively, overall small RNA (sRNA) biogenesis in A. hypogaea is markedly affected with an absence of the 24 nt sRNAs in TSWV-infected plants, possibly leading to wide-spread molecular and phenotypic perturbations specific to this species. These findings add significant information on the host–virus–vector interaction in terms of RNAi pathways and may lead to better crop and vector specific control strategies. PMID:27656190

  18. Pelacakan Virus Bercak Putih pada Udang Vaname (Litopenaeus vannamei di Lombok dengan Real-Time Polymerase Chain Reaction (DETECTION OF WHITE SPOT SYNDROME VIRUS IN LITOPENAEUS VANNAMEI IN LOMBOK ISLAND USING REAL-TIME POLYMERASE CHAIN REACTION

    Directory of Open Access Journals (Sweden)

    Lulu Arafani

    2016-03-01

    Full Text Available White spot syndrome virus (WSSV is one of the most threatening diseases in shrimp and othercrustaceans affecting global shrimp farming. Since firstly detected in Taiwan in 1992, the disease hasspread globally and followed with considerable socio-economic consequences. This research was performedto detect the WSSV infection in shrimp farming in Lombok Island’s (West Nusa Tenggara using real-timepolymerase chain reaction. Samples of vaname (Litopenaeus vannamei were collected from several shrimpfarming in Lombok. Results indicated that the spread of WSSV has reached shrimp farms in Lombok,especially in Lendang Jae, West Lombok. Therefore, a biosurveillance program is strongly recommendedto government to avoid and halt the spread of the disease in East Indonesia region .

  19. Reflective mulch and acibenzolar-S-methyl treatments relative to thrips (Thysanoptera: Thripidae) and tomato spotted wilt virus incidence in tomato.

    Science.gov (United States)

    Riley, D G; Joseph, S V; Srinivasan, R

    2012-08-01

    Management of thrips-transmitted tomato spotted wilt (TSW) virus typically relies on tactics that either reduce the thrips vector numbers or change the plant's response to the virus to reduce economic loss. We attempted to quantify the interaction between two such tactics, reflective mulch and the plant activator acibenzolar-S-methyl (Actigard), respectively, on a TSW-susceptible tomato hybrid. A split plot experiment was conducted in 2009 and 2010 where main-plots were three types of plastic mulch (two metalized reflective vs. black) and subplots consisted of a range of plant defense activator applications. TSW pressure varied over year with 80% of untreated plants having TSW in 2009 where as thrips and marketable yield in either year. In 2009, the seasonal average of Frankliniella fusca (Hinds) populations and incidence of TSW were significantly lower and yield significantly higher on both reflective mulches than on black mulch. Seasonal averages of thrips and fruit yield differed significantly among treatments of acibenzolar-S-methyl. However, there was a significant acibenzolar-S-methyl by mulch interaction relative to TSW incidence. In 2009, a minimum of acibenzolar-S-methyl at transplant plus foliar treatments at 10 and 20 d after transplant was required to significantly reduce TSW incidence compared with untreated plants before harvest. Under lower TSW pressure in 2010, average TSW incidence was significantly less in all plots treated with acibenzolar-S-methyl treated plots compared with the check. Acibenzolar-S-methyl treatments functioned better with the thrips reducing tactic, ultraviolet-reflective mulch. We propose that acibenzolar-S-methyl is less effective than metalized reflective mulch in reducing the incidence of TSW in tomato.

  20. Characterization of microRNAs by deep sequencing in red claw crayfish Cherax quadricarinatus haematopoietic tissue cells after white spot syndrome virus infection.

    Science.gov (United States)

    Zhao, Meng-Ru; Meng, Chuang; Xie, Xiao-Lu; Li, Cheng-Hua; Liu, Hai-Peng

    2016-12-01

    White spot syndrome virus (WSSV) is one of the most prevalent and widespread viruses in both shrimp and crayfish aquaculture. MicroRNAs (miRNAs) are crucial post-transcriptional regulators and play critical roles in cell differentiation and proliferation, apoptosis, signal transduction and immunity. In this study, miRNA expression profiles were identified via deep sequencing in red claw crayfish Cherax quadricarinatus haematopoietic tissue (Hpt) cell cultures infected with WSSV at both early (i.e., 1 hpi) and late (i.e., 12 hpi) infection stages. The results showed that 2 known miRNAs, namely, miR-7 and miR-184 play key roles in immunity. Meanwhile, 106 novel miRNA candidates were predicted by software in these combined miRNA transcriptomes. Compared with two control groups, 36 miRNAs showed significantly different expression levels after WSSV challenge. Furthermore, 10 differentially expressed miRNAs in WSSV-exposed Hpt cells were randomly selected for expression analysis by quantitative real-time RT-PCR. Consistent with the expression profiles identified by deep sequencing, RT-PCR showed a significant increase or decrease in miRNA expression in Hpt cells after WSSV infection. Prediction of targets of miRNAs such as miR-7, cqu-miR-52, cqu-miR-126 and cqu-miR-141 revealed that their target genes have diverse biological roles, including not only immunity but also transcriptional regulation, energy metabolism, cell communication, cell differentiation, cell death, autophagy, endocytosis and apoptosis. These results provide insight into the molecular mechanism of WSSV infection and highlight the function of miRNAs in the regulation of the immune response against WSSV infection in crustaceans. Copyright © 2016 Elsevier Ltd. All rights reserved.

  1. Manipulation of Frankliniella occidentalis (Thysanoptera: Thripidae) by Tomato Spotted Wilt Virus (Tospovirus) Via the Host Plant Nutrients to Enhance Its Transmission and Spread.

    Science.gov (United States)

    Shalileh, Sheida; Ogada, Pamella Akoth; Moualeu, Dany Pascal; Poehling, Hans-Michael

    2016-10-01

    Earlier studies have shown that Tomato spotted wilt virus (TSWV) influences the biology, performance, and behavioral patterns of its vector Frankliniella occidentalis Pergande. In this study, using Capsicum annuum L. as the host plant, we aimed to determine the manipulation of F. occidentalis by TSWV through switching of the diet (+ or -TSWV) during vector's development. Behavioral patterns, fitness, as well as vector performance were evaluated. The specific parameters investigated included longevity/survival, fecundity, development time, feeding, and preferential behavior. F. occidentalis were reared on either TSWV-infected (exposed) or healthy leaves (non-exposed) throughout their larval stages. The emerging adults were then individually transferred to either healthy or TSWV-infected leaf disks. This resulted into four treatments, consisting of exposed or non-exposed thrips reared on either infected or healthy leaf disks as adults. All F. occidentalis exposed to TSWV in their larval stages had shorter development time regardless of the adults' diet. Whereas, the ones that were later reared on healthy leaf disks as adults recorded the highest longevity and reproduction rate. Furthermore, adults of F. occidentalis that were exposed to TSWV in their larval stages showed preference toward healthy leaf disks (-TSWV), whereas the non-exposed significantly preferred the infected leaf disks (+TSWV). These are further indications that TSWV modifies the nutritional content of its host plants, which influences vector's biology and preferential behavior, in favor of its multiplication and dispersal. The findings offer additional explanation to the often aggressive spread of the virus in crop stands. © The Authors 2016. Published by Oxford University Press on behalf of Entomological Society of America.

  2. Responses of three very large inducible GTPases to bacterial and white spot syndrome virus challenges in the giant fresh water prawn Macrobrachium rosenbergii.

    Science.gov (United States)

    Huang, Ying; Jin, Min; Yin, Shaowu; Ding, Zhengfeng; Wang, Wen; Ren, Qian

    2016-04-01

    Interferons (IFNs) are cytokines secreted by cells in response to invasion by pathogens, such as viruses, bacteria, parasites, or tumor cells. Very large inducible GTPases (VLIG) are the latest IFN-inducible GTPase family to be discovered and are the largest known GTPases of any species. However, VLIG proteins from invertebrates have yet to be characterized. In this study, three forms of VLIGs designated as MrVLIG1, MrVLIG2, and MrVLIG3 were cloned from the giant fresh water prawn Macrobrachium rosenbergii. MrVLIG1 has a 5445 bp open reading frame (ORF) encoding an 1814-amino acid protein. The complete nucleotide sequence of MrVLIG2 cDNA is 7055 bp long consisting of a 5757 bp ORF encoding a protein with 1918 amino acids. The full length of the MrVLIG3 gene consists of 5511 bp with a 3909 bp ORF encoding a peptide with 1302 amino acids. BLASTP and phylogenetic tree analyses showed that the three MrVLIGs are clustered into one subgroup and, together with other vertebrate VLIGs, into a branch. Tissue distribution analysis indicated that the mRNAs of the three MrVLIGs were widely expressed in almost all detected tissues, including the hemocytes, heart, hepatopancreas, gills, stomach, and intestine, with the highest expression in the hepatopancreas. They were also detected in the intestine but with relatively low expression levels. Quantitative real-time RT-PCR analysis showed that the mRNA transcripts of the MrVLIGs in the hepatopancreas were significantly expressed at various time points after infection with Vibrio parahaemolyticus and white spot syndrome virus. In summary, the three isoforms of VLIG genes participate in the innate immune response of the shrimps to bacterial and viral infections. Copyright © 2016 Elsevier Ltd. All rights reserved.

  3. Spotted Lanternfly

    OpenAIRE

    Sisti, Phillip A.; Pfeiffer, Douglas G.; Day, Eric R.

    2016-01-01

    Describes the Spotted Lanternfly (Lycorma delicatula), its life cycle, and potential for invasive spread in North America. At present this species is known in the US only from four counties in Pennsylvania, where there is a quarantine.

  4. Identification of a novel shrimp protein phosphatase and its association with latency-related ORF427 of white spot syndrome virus.

    Science.gov (United States)

    Lu, Liqun; Kwang, Jimmy

    2004-11-05

    To characterize the role of latency-associated ORF427 of white spot syndrome virus (WSSV), a shrimp cDNA library was constructed to screen interacting proteins of ORF427. Employing the yeast two-hybrid system, a novel shrimp protein phosphatase (named PPs), sharing 93% homology with human protein phosphatase 1, has been identified able to bind ORF427 in yeast. Through co-immunoprecipitation assays, the interaction between PPs and ORF427 was further confirmed both in vitro and in vivo. Interestingly, the novel shrimp protein phosphatase consists of only 199 aa and contains almost all the functional catalytic domains of human protein phosphatase, while it lacks the corresponding C-terminal non-catalytic sequence. Transcription and translation products of the identified cDNA can be detected in both normal and WSSV-infected shrimps; and PPs was found to localize mainly in the lysosome of shrimp cells. To characterize its function, the PPs cDNA was highly expressed in bacteria and the purified protein showed phosphatase activity when tested against pNPP in a standard phosphatase assay. Our results suggest that the identified protein phosphatase, PPs, may represent a novel member of protein phosphatase family and might be involved in the regulation of WSSV's life cycle through interaction with latency-related ORF427 of WSSV.

  5. The concentration of white spot disease virus for its detection in sea water using a combined ferric colloid adsorption- and foam separation-based method.

    Science.gov (United States)

    Suzuki, Yoshihiro; Suzuki, Takahiko; Kono, Tomoya; Mekata, Tohoru; Sakai, Masahiro; Itami, Toshiaki

    2011-05-01

    The white spot disease virus (WSDV), which is most virulent in shrimp, is a cause of serious damage in the shrimp production industry. However, it is difficult to track the infection route and behaviour of WSDV in shrimp farms because it is present at extremely low concentrations in culture sea water. In this study, the concentration of WSDV in sea water foam was examined using dispersed bubbles and milk casein as a surface-active protein. WSDV concentrations were assessed using real-time PCR. When ferric colloid adsorption was performed prior to foam separation, WSDV was effectively removed from sea water and concentrated in the generated foam within 5 min. The removal efficiency was greater than 90% at the optimum iron and casein concentrations of 1mg Fe/l and 1mg/l, respectively. Furthermore, to analyse the dissolution of the collected ferric colloid, the WSDV concentration in the colloid-dissolved solution was set to be 200-fold higher than that found in raw water. This represents a novel method of concentrating WSDV for its detection in sea water using a combination of ferric colloid adsorption and foam separation that is easy to perform, rapid and efficient. Copyright © 2011 Elsevier B.V. All rights reserved.

  6. Indel-II region deletion sizes in the white spot syndrome virus genome correlate with shrimp disease outbreaks in southern Vietnam.

    Science.gov (United States)

    Hoa, Tran Thi Tuyet; Zwart, Mark P; Phuong, Nguyen T; Oanh, Dang T H; de Jong, Mart C M; Vlak, Just M

    2012-06-13

    Sequence comparisons of the genomes of white spot syndrome virus (WSSV) strains have identified regions containing variable-length insertions/deletions (i.e. indels). Indel-I and Indel-II, positioned between open reading frames (ORFs) 14/15 and 23/24, respectively, are the largest and the most variable. Here we examined the nature of these 2 indel regions in 313 WSSV-infected Penaeus monodon shrimp collected between 2006 and 2009 from 76 aquaculture ponds in the Mekong Delta region of Vietnam. In the Indel-I region, 2 WSSV genotypes with deletions of either 5950 or 6031 bp in length compared with that of a reference strain from Thailand (WSSV-TH-96-II) were detected. In the Indel-II region, 4 WSSV genotypes with deletions of 8539, 10970, 11049 or 11866 bp in length compared with that of a reference strain from Taiwan (WSSV-TW) were detected, and the 8539 and 10970 bp genotypes predominated. Indel-II variants with longer deletions were found to correlate statistically with WSSV-diseased shrimp originating from more intensive farming systems. Like Indel-I lengths, Indel-II lengths also varied based on the Mekong Delta province from which farmed shrimp were collected.

  7. Complete genome sequences of three tomato spotted wilt virus isolates from tomato and pepper plants in Korea and their phylogenetic relationship to other TSWV isolates.

    Science.gov (United States)

    Lee, Jong-Seung; Cho, Won Kyong; Kim, Mi-Kyeong; Kwak, Hae-Ryun; Choi, Hong-Soo; Kim, Kook-Hyung

    2011-04-01

    Tomato spotted wilt virus (TSWV) infects numerous host plants and has three genome segments, called L, M and S. Here, we report the complete genome sequences of three Korean TSWV isolates (TSWV-1 to -3) infecting tomato and pepper plants. Although the nucleotide sequence of TSWV-1 genome isolated from tomato is very different from those of TSWV-2 and TSWV-3 isolated from pepper, the deduced amino acid sequences of the five TSWV genes are highly conserved among all three TSWV isolates. In phylogenetic analysis, deduced RdRp protein sequences of TSWV-2 and TSWV-3 were clustered together with two previously reported isolates from Japan and Korea, while TSWV-1 grouped together with a Hawaiian isolate. A phylogenetic tree based on N protein sequences, however, revealed four distinct groups of TSWV isolates, and all three Korean isolates belonged to group II, together with many other isolates, mostly from Europe and Asia. Interestingly, most American isolates grouped together as group I. Together, these results suggested that these newly identified TSWV isolates might have originated from an Asian ancestor and undergone divergence upon infecting different host plants.

  8. White spot syndrome virus entry is dependent on multiple endocytic routes and strongly facilitated by Cq-GABARAP in a CME-dependent manner

    Science.gov (United States)

    Chen, Rong-yuan; Shen, Kai-li; Chen, Zhen; Fan, Wei-wei; Xie, Xiao-lu; Meng, Chuang; Chang, Xue-jiao; Zheng, Li-bing; Jeswin, Joseph; Li, Cheng-hua; Wang, Ke-jian; Liu, Hai-peng

    2016-01-01

    White spot syndrome virus (WSSV) is a lethal pathogen of shrimp and many other crustaceans, including crayfish. However, the molecular mechanism underlying its cellular entry remains elusive due to the lack of shrimp cell lines for viral propagation. Crayfish hematopoietic tissue (Hpt) cell culture was recently established as a good model for WSSV infection study. Here, we showed that multiple endocytic routes, including clathrin-mediated endocytosis (CME), macropinocytosis and caveolae-mediated endocytosis, were indispensably employed for the viral entry into Hpt cell of the crayfish Cherax quadricarinatus. Intriguingly, cellular autophagic activity was positively correlated with efficient viral entry, in which a key autophagy-related protein, γ-aminobutyric acid receptor-associated protein (Cq-GABARAP), that not only localized but also co-localized with WSSV on the Hpt cell membrane, strongly facilitated WSSV entry by binding to the viral envelope VP28 in a CME-dependent manner that was negatively regulated by Cq-Rac1. Furthermore, cytoskeletal components, including Cq-β-tubulin and Cq-β-actin, bound to both recombinant rCq-GABARAP and WSSV envelope proteins, which likely led to viral entry promotion via cooperation with rCq-GABARAP. Even under conditions that promoted viral entry, rCq-GABARAP significantly reduced viral replication at an early stage of infection, which was probably caused by the formation of WSSV aggregates in the cytoplasm. PMID:27385304

  9. Proteomic analysis by iTRAQ in red claw crayfish, Cherax quadricarinatus, hematopoietic tissue cells post white spot syndrome virus infection.

    Science.gov (United States)

    Jeswin, Joseph; Xie, Xiao-lu; Ji, Qiao-lin; Wang, Ke-jian; Liu, Hai-peng

    2016-03-01

    To elucidate proteomic changes of Hpt cells from red claw crayfish, Cherax quadricarinatus, we have carried out isobaric tags for relative and absolute quantitation (iTRAQ) of cellular proteins at both early (1 hpi) and late stage (12 hpi) post white spot syndrome virus (WSSV) infection. Protein database search revealed 594 protein hits by Mascot, in which 17 and 30 proteins were present as differentially expressed proteins at early and late viral infection, respectively. Generally, these differentially expressed proteins include: 1) the metabolic process related proteins in glycolysis and glucogenesis, DNA replication, nucleotide/amino acid/fatty acid metabolism and protein biosynthesis; 2) the signal transduction related proteins like small GTPases, G-protein-alpha stimulatory subunit, proteins bearing PDZ- or 14-3-3-domains that help holding together and organize signaling complexes, casein kinase I and proteins of the MAP-kinase signal transduction pathway; 3) the immune defense related proteins such as α-2 macroglobulin, transglutaminase and trans-activation response RNA-binding protein 1. Taken together, these protein information shed new light on the host cellular response against WSSV infection in a crustacean cell culture. Copyright © 2016 Elsevier Ltd. All rights reserved.

  10. Identification and characterization of intestine microRNAs and targets in red swamp crayfish, Procambarus clarkii infected with white spot syndrome virus.

    Directory of Open Access Journals (Sweden)

    Zhi-Qiang Du

    Full Text Available MicroRNAs (miRNAs are small non-coding endogenous RNA molecules that play important roles in the innate immunity system of invertebrates, especially in the aspect of antivirus. In the present study, high-throughput small RNA Illumina sequencing systems were used to identify differentially expressed miRNAs (DEMs from the intestines of Procambarus clarkii that were infected with white spot syndrome virus (WSSV. As a result, 39 known and 12 novel miRNAs were identified in both NG and WG small RNA libraries. Seven DEMs were determined to be involved in the antiviral innate immunity in the intestines of P. clarkii. The results of the target gene predictions of the DEMs showed that the putative target genes of these 7 DEMs are related to tight junctions, vascular smooth muscle contraction regulation of the actin cytoskeleton, focal adhesion, RNA transport, mRNA surveillance, viral carcinogenesis, and Salmonella infection. These results provide theoretical insights for future studies on the antiviral immunity of crustaceans.

  11. Peritrophin-like protein from Litopenaeus vannamei (LvPT) involved in white spot syndrome virus (WSSV) infection in digestive tract challenged with reverse gavage

    Science.gov (United States)

    Xie, Shijun; Li, Fuhua; Zhang, Xiaojun; Zhang, Jiquan; Xiang, Jianhai

    2017-05-01

    The peritrophic membrane plays an important role in the defense system of the arthropod gut. The digestive tract is considered one of the major tissues targeted by white spot syndrome virus (WSSV) in shrimp. In this study, the nucleotide sequence encoding peritrophin-like protein of Litopenaeus vannamei (LvPT) was amplified from a yeast two-hybrid library of L. vannamei. The epitope peptide of LvPT was predicted with the GenScript OptimumAntigen™ design tool. An anti-LvPT polyclonal antibody was produced and shown to specifically bind a band at 27 kDa, identified as LvPT. The LvPT protein was expressed and its concentration determined. LvPT dsRNA (4 μg per shrimp) was used to inhibit LvPT expression in shrimp, and a WSSV challenge experiment was then performed with reverse gavage. The pleopods, stomachs, and guts were collected from the shrimp at 0, 24, 48, and 72 h post-infection (hpi). Viral load quantification showed that the levels of WSSV were significantly lower in the pleopods, stomachs, and guts of shrimp after LvPT dsRNA interference than in those of the controls at 48 and 72 hpi. Our results imply that LvPT plays an important role during WSSV infection of the digestive tract.

  12. Peritrophin-like protein from Litopenaeus vannamei (LvPT) involved in white spot syndrome virus (WSSV) infection in digestive tract challenged with reverse gavage

    Science.gov (United States)

    Xie, Shijun; Li, Fuhua; Zhang, Xiaojun; Zhang, Jiquan; Xiang, Jianhai

    2017-11-01

    The peritrophic membrane plays an important role in the defense system of the arthropod gut. The digestive tract is considered one of the major tissues targeted by white spot syndrome virus (WSSV) in shrimp. In this study, the nucleotide sequence encoding peritrophin-like protein of Litopenaeus vannamei (LvPT) was amplified from a yeast two-hybrid library of L. vannamei. The epitope peptide of LvPT was predicted with the GenScript OptimumAntigen™ design tool. An anti-LvPT polyclonal antibody was produced and shown to specifically bind a band at 27 kDa, identified as LvPT. The LvPT protein was expressed and its concentration determined. LvPT dsRNA (4 μg per shrimp) was used to inhibit LvPT expression in shrimp, and a WSSV challenge experiment was then performed with reverse gavage. The pleopods, stomachs, and guts were collected from the shrimp at 0, 24, 48, and 72 h post-infection (hpi). Viral load quantification showed that the levels of WSSV were significantly lower in the pleopods, stomachs, and guts of shrimp after LvPT dsRNA interference than in those of the controls at 48 and 72 hpi. Our results imply that LvPT plays an important role during WSSV infection of the digestive tract.

  13. Complete nucleotide sequences of the genomes of two isolates of apple chlorotic leaf spot virus from peach (Prunus persica) in China.

    Science.gov (United States)

    Niu, Feiqing; Pan, Song; Wu, Zujian; Jiang, Dongmei; Li, Shifang

    2012-04-01

    The complete nucleotide sequences of two isolates of apple chlorotic leaf spot virus (Z1 and Z3) collected from peach in Henan Province, China, were determined. The genomes of both Z1 and Z3 were found to contain three open reading frames (ORFs). Sequence analysis showed that genomic sequences of Z1 and Z3 isolates shared 67.4%-82.9% and 67.2%-82.6% identity, respectively, with the other eight isolates of ACLSV that have been reported previously. Based on the putative amino acid sequences of the products of the three ORFs, Z1 and Z3 isolates showed the greatest identity to isolate PBM1 (GenBank accession number AJ243438) from plum and the least identity with isolate Ta Tao5 (GenBank Accession Number: EU223295) from peach. Considering the low level of sequence identity between Z1/Z3 isolate and Ta Tao5 isolate, two types of ACLSV may exist in peach.

  14. Transmission of white spot syndrome virus (WSSV) to Litopenaeus vannamei from infected cephalothorax, abdomen, or whole shrimp cadaver.

    Science.gov (United States)

    Soto, M A; Shervette, V R; Lotz, J M

    2001-06-20

    Shrimp viruses can remain infectious in frozen shrimp tissue and have been found in frozen commodity shrimp. Therefore, the threat of viral outbreaks in wild and cultured shrimp via frozen commodity shrimp exists. Because frozen shrimp are imported with and without the cephalothorax, more knowledge is needed concerning the infectivity of a cephalothorax relative to that of an abdomen. We compared the mortality rates from shrimp exposed to a WSSV-infected cephalothorax, abdomen, or whole shrimp cadaver. Estimates of transmission coefficients from the exposures to the infected cephalothorax, abdomen, or whole shrimp were also calculated because the transmission coefficients account for differences in the initial doses. In addition, we compared the variability in infectivity of pieces of shrimp by feeding 24 equal-sized pieces of cephalothorax and abdomen to 24 individually isolated shrimp. In Expt 1, susceptible shrimp did not completely consume the infected abdomen, and a significant difference was detected among shrimp exposed to the abdomen (mortality rate = 0.40), cephalothorax (mortality rate = 0.75), and whole shrimp cadaver (mortality rate = 0.67). The calculated transmission coefficients were 0.95 from an infected cephalothorax, 0.59 from an infected abdomen, and 0.69 from an infected whole shrimp cadaver. In Expt 2, susceptible shrimp were starved to ensure complete ingestion of each dose. No significant difference was observed in the estimated mortality rates from an infected cephalothorax (0.58), abdomen (0.63), or whole shrimp (0.67). The calculated transmission coefficients were 0.84 from an infected cephalothorax, 0.83 from an infected abdomen, and 0.60 from an infected whole shrimp cadaver. In Expt 3, no difference was observed in the mortality rates resulting from exposures to pieces of infected cephalothorax (0.57) or abdomen (0.58). Our results suggested that there was no difference in the viral loads of a WSSV-infected cephalothorax or abdomen, but

  15. Atividade predatória de isolados de Arthrobotrys spp sobre larvas infectantes de Cooperia punctata

    Directory of Open Access Journals (Sweden)

    Jackson Victor de Araújo

    1998-01-01

    Full Text Available Experimentos laboratoriais foram realizados para investigar a capacidade de isolados de fungos predadores das espécies Arthrobotrys musiformis (isolado 3, A. conoides (isolado A e A. robusta (isolados B e E de predar e matar larvas infectantes de Cooperia punctata. Dois grupos foram formados para o teste de cada isolado: grupo 1, fungos e larvas infectantes e grupo 2, larvas infectantes (controle. Houve diferença estatisticamente significativa (p;0,05 foi encontrada entre o isolado E e o grupo controle. Isto pode indicar uma variação existente dentro de uma mesma espécie de fungo ou gênero quanto à predação de larvas infectantes de C. punctata.

  16. Development of transgenic papayas expressing the coat protein gene from a Brazilian isolate of Papaya ringspot virus (PRSV) = Desenvolvimento de mamoeiros transgênicos resistentes a vírus expressando o gene da capa protéica de um isolado brasileiro de Papaya ringspot virus

    NARCIS (Netherlands)

    Souza, M.T.; Níckel, O.; Gonsalves, D.

    2005-01-01

    Translatable and nontranslatable versions of the coat protein (cp) gene of a Papaya ringspot virus (PRSV) isolate collected in the state of Bahia, Brazil, were engineered for expression in Sunrise and Sunset Solo varieties of papaya (Carica papaya). The biolistic system was used to transform

  17. Global analysis of population structure, spatial and temporal dynamics of genetic diversity, and evolutionary lineages of Iris yellow spot virus (Tospovirus: Bunyaviridae).

    Science.gov (United States)

    Iftikhar, Romana; Ramesh, Shunmugiah V; Bag, Sudeep; Ashfaq, Muhammad; Pappu, Hanu R

    2014-08-15

    Thrips-transmitted Iris yellow spot virus is an economically important viral pathogen of Allium crops worldwide. A global analysis of known IYSV nucleocapsid gene (N gene) sequences was carried out to determine the comparative population structure, spatial and temporal dynamics with reference to its genetic diversity and evolution. A total of 98 complete N gene sequences (including 8 sequences reported in this study) available in GenBank and reported from 23 countries were characterized by in-silico RFLP analysis. Based on RFLP, 94% of the isolates could be grouped into NL or BR types while the rest belonged to neither group. The relative proportion of NL and BR types was 46% and 48%, respectively. A temporal shift in the IYSV genotypes with a greater incremental incidence of IYSVBR was found over IYSVNL before 2005 compared to after 2005. The virus population had at least one evolutionarily significant recombination event, involving IYSVBR and IYSVNL. Codon substitution studies did not identify any significant differences among the genotypes of IYSV. However, N gene codons were minimally positively selected, moderately negatively selected denoting the action of purifying selection, thus rejecting the theory of neutral mutation in IYSV population. However, one codon position (139) was found to be positively selected in all the genotypes. Population selection statistics in the IYSVBR, IYSVNL genotypes and in the population as a whole also revealed the action of purifying selection or population expansion, whereas IYSVother displayed a decrease in population size. Genetic differentiation studies showed inherent differentiation and infrequent gene flow between IYSVBR and IYSVNL genotypes corroborating the geographical confinement of these genotypes. Taken together the study suggests that the observed diversity in IYSV population and temporal shift in IYSVBR genotype is attributable to genetic recombination, abundance of purifying selection, insignificant positive

  18. Role of Litopenaeus vannamei Yin Yang 1 in the Regulation of the White Spot Syndrome Virus Immediate Early Gene ie1.

    Science.gov (United States)

    Huang, Ping-Han; Huang, Ting-Yi; Cai, Pei-Si; Chang, Li-Kwan

    2017-03-15

    Yin Yang 1 (YY1) is a multifunctional zinc finger transcription factor that regulates many key cellular processes. In this study, we report the cloning of YY1 from Litopenaeus vannamei shrimp (LvYY1). This study shows that LvYY1 is ubiquitously expressed in shrimp tissues, and knockdown of LvYY1 expression by double-stranded RNA (dsRNA) injection in white spot syndrome virus (WSSV)-infected shrimp reduced both mRNA levels of the WSSV immediate early gene ie1 as well as overall copy numbers of the WSSV genome. The cumulative mortality rate of infected shrimp also declined with LvYY1 dsRNA injection. Using an insect cell model, we observed that LvYY1 activates ie1 expression, and a mutation introduced into the ie1 promoter subsequently repressed this capability. Moreover, reporter assay results suggested that LvYY1 is involved in basal transcriptional regulation via an interaction with L. vannamei TATA-binding protein (LvTBP). Electrophoretic mobility shift assay (EMSA) results further indicated that LvYY1 binds to a YY1-binding site in the region between positions -119 and -126 in the ie1 promoter. Chromatin immunoprecipitation analysis also confirmed that LvYY1 binds to the ie1 promoter in WSSV-infected shrimp. Taken together, these results indicate that WSSV uses host LvYY1 to enhance ie1 expression via a YY1-binding site and the TATA box in the ie1 promoter, thereby facilitating lytic activation and viral replication.IMPORTANCE WSSV has long been a scourge of the shrimp industry and remains a serious global threat. Thus, there is a pressing need to understand how the interactions between WSSV and its host drive infection, lytic development, pathogenesis, and mortality. Our successful cloning of L. vannamei YY1 (LvYY1) led to the elucidation of a critical virus-host interaction between LvYY1 and the WSSV immediate early gene ie1 We observed that LvYY1 regulates ie1 expression via a consensus YY1-binding site and TATA box. LvYY1 was also found to interact with L

  19. The c-Fos and c-Jun from Litopenaeus vannamei play opposite roles in Vibrio parahaemolyticus and white spot syndrome virus infection.

    Science.gov (United States)

    Li, Chaozheng; Li, Haoyang; Wang, Sheng; Song, Xuan; Zhang, Zijian; Qian, Zhe; Zuo, Hongliang; Xu, Xiaopeng; Weng, Shaoping; He, Jianguo

    2015-09-01

    Growing evidence indicates that activator protein-1 (AP-1) plays a major role in stimulating the transcription of immune effector molecules in cellular response to an incredible array of stimuli, including growth factors, cytokines, cellular stresses and bacterial and viral infection. Here, we reported the isolation and characterization of a cDNA from Litopenaeus vannamei encoding the full-length c-Fos protein (named as Lvc-Fos). The predicted amino acid sequences of Lvc-Fos contained a basic-leucine zipper (bZIP) domain, which was characteristic of members of the AP-1 family. Immunoprecipitation and native-PAGE assays determined that Lvc-Fos could interact with the Lvc-Jun, a homolog of c-Jun family in L. vannamei, in a heterodimer manner. Further investigation demonstrated that Lvc-Fos and Lvc-Jun were expressed in all tested tissues and located in the nucleus. Real-time RT-PCR analysis showed both Lvc-Fos and Lvc-Jun in gills were up-regulated during Vibrio parahaemolyticus and white spot syndrome virus (WSSV) challenges. In addition, reporter gene assays indicated Lvc-Fos and Lvc-Jun could activate the expression of antimicrobial peptides (AMPs) of Drosophila and shrimp, as well as WSSV immediate early (IE) genes wsv069 and wsv249, in a different manner. Knockdown of Lvc-Fos or Lvc-Jun by RNA interference (RNAi) resulted in higher mortalities of L. vannamei after infection with V. parahaemolyticus, suggesting that Lvc-Fos and Lvc-Jun might play protective roles in bacterial infection. However, silencing of Lvc-Fos or Lvc-Jun in shrimp caused lower mortalities and virus loads under WSSV infection, suggesting that Lvc-Fos and Lvc-Jun could be engaged for WSSV replication and pathogenesis. In conclusion, our results provided experimental evidence and novel insight into the roles of L. vannamei AP-1 in bacterial and viral infection. Copyright © 2015 Elsevier Ltd. All rights reserved.

  20. Analysis of variable genomic loci in white spot syndrome virus to predict its origins in Procambarus clarkii crayfish farmed in China.

    Science.gov (United States)

    Zeng, Wei; Zeng, Yong; Fei, Rong-Mei; Zeng, Ling-Bing; Wei, Kai-Jin

    2011-09-09

    Variable genomic loci were examined in 4 white spot syndrome virus (WSSV) isolates (08HB, 09HB, 08JS and 09JS) from Procambarus clarkii crayfish collected from Jiangsu and Hubei Provinces in China in 2008 and 2009. In ORF75, sequence variation detected in the 4 isolates, as well as in isolates sequenced previously, suggested that WSSV might have segregated into 2 lineages since first emerging as a serious pathogen of farmed shrimp in East Asia in the early-mid 1990s, with one lineage remaining in East Asia and the other separating to South Asia. In ORF23/24, deletions of 9.31, 10.97, or 11.09 kb were evident compared to a reference isolate from Taiwan (WSSV-TW), and, in ORF14/15, deletions of 5.14 or 5.95 kb were evident compared to a reference isolate from Thailand with the largest genome size (TH-96-II). With respect to these genome characteristics, the crayfish isolates 08HB, 09HB and 08JS were similar to WSSV-TW and the isolate 09JS was similar to a reference isolate from China (WSSV-CN). In addition to these loci, sequence variation was evident in ORF94 and ORF125 that might be useful for differentiating isolates and in epidemiological tracing of WSSV spread in crayfish farmed in China. However, as all 4 crayfish isolates possessed a Homologous Region 9 sequence identical to isolate WSSV-TW and another Thailand isolate (WSSV-TH), and as their transposase sequence was identical to isolates WSSV-CN and WSSV-TH, these 2 loci were not useful in predicting their origins.

  1. A laminin-receptor-like protein regulates white spot syndrome virus infection by binding to the viral envelope protein VP28 in red claw crayfish Cherax quadricarinatus.

    Science.gov (United States)

    Liu, Ling-Ke; Li, Wei-Dong; Gao, Yan; Chen, Rong-Yuan; Xie, Xiao-Lu; Hong, Heng; Wang, Ke-Jian; Liu, Hai-Peng

    2018-02-01

    White spot syndrome virus (WSSV) is a lethal pathogen of shrimp and many other crustaceans, which has been causing huge economic losses in global aquaculture. Laminin receptor (LR) is a cell surface receptor which participates in the interactions between cells as well as cells and extracellular matrix. Previously, we found that a CqLR-like gene was responsive to WSSV infection in the hematopoietic tissue (Hpt) cells from red claw crayfish Cherax quadricarinatus. To further reveal the role of CqLR-like gene involved in WSSV infection, the full-length cDNA of CqLR-like gene was cloned with 1000 bp, and the open reading frame encoded 308 amino acids with a conserved laminin-binding domain. Importantly, both the WSSV entry and viral replication were strongly reduced in Hpt cells after loss-of-function of CqLR-like gene by gene silencing. Protein interaction assay demonstrated that the recombinant CqLR-like protein could bind to WSSV virion in vitro by enzyme-linked immunosorbent assay and the binding affinity was in a dose-dependent manner. Furthermore, recombinant CqLR-like protein was found to bind to WSSV envelop protein VP28, but not other envelop proteins tested including VP19, VP24, and VP26, by pull down assay in HEK293T cells. In regarding to that LR is mainly localized on many types of cells' membrane, these data together suggested that CqLR-like protein was likely to function as a putative recognition molecule towards WSSV and act in the viral entry into a crustacean host cell, which may benefit the elucidation of the WSSV pathogenesis and further the pharmaceutical target for the possibly effective control of WSSV disease. Copyright © 2017 Elsevier Ltd. All rights reserved.

  2. CqToll participates in antiviral response against white spot syndrome virus via induction of anti-lipopolysaccharide factor in red claw crayfish Cherax quadricarinatus.

    Science.gov (United States)

    Li, Yan-Yao; Chen, Xiao-Xiao; Lin, Feng-Yu; Chen, Qiu-Fan; Ma, Xing-Yuan; Liu, Hai-Peng

    2017-09-01

    It is well known that Tolls/Toll like receptors (TLRs), a family of pattern recognition receptors, play important roles in immune responses. Previously, we found that a Toll transcript was increased in a transcriptome library of haematopoietic tissue (Hpt) cells from the red claw crayfish Cherax quadricarinatus post white spot syndrome virus infection. In the present study, a full-length cDNA sequence of Toll receptor (named as CqToll) was identified with 3482 bp which contained an open reading frame of 3021 bp encoding 1006 amino acids. The predicted structure of CqToll protein was composed of three domains, including an extracellular domain of 19 leucine-rich repeats residues, a transmembrane domain and an intracellular domain of 138 amino acids. Tissue distribution analysis revealed that CqToll was expressed widely in various tissues determined from red claw crayfish with highest expression in haemocyte but lowest expression in eyestalk. Importantly, significant lower expression of the anti-lipopolysacchride factor (CqALF), an antiviral antimicrobial peptide (AMP) in crustaceans, but not CqCrustin was observed after gene silencing of CqToll in crayfish Hpt cell cultures, indicating that the CqALF was likely to be positively regulated via Toll pathway in red claw crayfish. Furthermore, the transcription of both an immediate early gene and a late envelope protein gene VP28 of WSSV were clearly enhanced in Hpt cells if silenced with CqToll, suggesting that the increase of WSSV replication was likely to be caused by the lower expression of the CqALF resulted from the loss-of-function of CqToll. Taken together, these data implied that CqToll might play a key role in anti-WSSV response via induction of CqALF in a crustacean C. quadricarinatus. Copyright © 2017. Published by Elsevier Ltd.

  3. Litopenaeus vannamei tumor necrosis factor receptor-associated factor 6 (TRAF6) responds to Vibrio alginolyticus and white spot syndrome virus (WSSV) infection and activates antimicrobial peptide genes.

    Science.gov (United States)

    Wang, Pei-Hui; Wan, Ding-Hui; Gu, Zhi-Hua; Deng, Xie-Xiong; Weng, Shao-Ping; Yu, Xiao-Qiang; He, Jian-Guo

    2011-01-01

    Tumor necrosis factor receptor (TNFR)-associated factor 6 (TRAF6) is a key signaling adaptor protein not only for the TNFR superfamily but also for the Interleukin-1 receptor/Toll-like receptor (IL-1/TLR) superfamily. To investigate TRAF6 function in invertebrate innate immune responses, Litopenaeus vannamei TRAF6 (LvTRAF6) was identified and characterized. The full-length cDNA of LvTRAF6 is 2823bp long, with an open reading frame (ORF) encoding a putative protein of 594 amino acids, including a RING-type Zinc finger, two TRAF-type Zinc fingers, a coiled-coil region, and a meprin and TRAF homology (MATH) domain. The overall amino acid sequence identity between LvTRAF6 and other known TRAF6s is 22.2-33.3%. Dual luciferase reporter assays in Drosophila S2 cells revealed that LvTRAF6 could activate the promoters of antimicrobial peptide genes (AMPs), including Drosophila Attacin A and Drosomycin, and shrimp Penaeidins. Real-time quantitative PCR (qPCR) indicated that LvTRAF6 was constitutively expressed in various tissues of L. vannamei. After Vibrio alginolyticus and white spot syndrome virus (WSSV) challenge, LvTRAF6 was down-regulated, though with different expression patterns in the intestine compared to other tissues. After WSSV challenge, LvTRAF6 was up-regulated 2.7- and 2.3-fold over the control at 3h in gills and hepatopancreas, respectively. These results indicated that LvTRAF6 may play a crucial role in antibacterial and antiviral responses via regulation of AMP gene expression. Copyright © 2010 Elsevier Ltd. All rights reserved.

  4. Role of insecticides in reducing thrips injury to plants and incidence of tomato spotted wilt virus in Virginia market-type peanut.

    Science.gov (United States)

    Herbert, D Ames; Malone, S; Aref, S; Brandenburg, R L; Jordan, D L; Royals, B M; Johnson, P D

    2007-08-01

    Tomato spotted wilt virus (family Bunyaviridae, genus Tospovirus, TSWV), transmitted by many thrips species, is a devastating pathogen of peanut, Arachis hypogaea L. TSWV has become a serious problem in the Virginia/Carolina peanut-growing region of the United States. During 2002, TSWV was present in 47% of the North Carolina hectarage and caused a 5% yield reduction in Virginia. Factors influencing levels of TSWV in runner market-type peanut cultivars, which are primarily grown in Alabama, Flordia, Georgia, and Texas, have been integrated into an advisory to help those peanut growers reduce losses. An advisory based on the southeast runner market-type version is currently under development for virginia market-type peanut cultivars that are grown primarily in the Virginia/ Carolina region. A version based on preliminary field experiments was released in 2003. One factor used in both advisories relates to insecticide use to reduce the vector populations and disease incidence. This research elucidated the influence of insecticides on thrips populations, thrips plant injury, incidence of TSWV, and pod yield in virginia market-type peanut. Eight field trials from 2003 to 2005 were conducted at two locations. In-furrow application of aldicarb and phorate resulted in significant levels of thrips control, significant reductions in thrips injury to seedlings, reduced incidence of TSWV, and significant increases in pod yield. Foliar application of acephate after aldicarb or phorate applied in the seed furrow further reduced thrips plant injury and incidence of TSWV and improved yield. These findings will be used to improve the current virginia market-type TSWV advisory.

  5. Long-Distance Dispersal Potential for Onion Thrips (Thysanoptera: Thripidae) and Iris yellow spot virus (Bunyaviridae: Tospovirus) in an Onion Ecosystem.

    Science.gov (United States)

    Smith, Erik A; Fuchs, M; Shields, E J; Nault, B A

    2015-08-01

    Onion thrips, Thrips tabaci Lindeman, is a worldwide pest of onion whose feeding damage and transmission of Iris yellow spot virus (IYSV) may reduce onion yields. Little is known about the seasonal dynamics of T. tabaci dispersal, the distance of dispersal, or the movement of thrips infected with IYSV during the onion-growing season. To address these questions, T. tabaci adults were collected using transparent sticky card traps in commercial onion fields three times during the onion-growing season (June, July, and late August) at varying heights above the canopy (0.5-6 m above soil surface) and with trap-equipped unmanned aircraft (UAVs) flying 50-60 m above onion fields during August sampling periods in 2012 and 2013. Randomly selected subsamples of captured T. tabaci were tested for IYSV using RT-PCR. Most T. tabaci adults were captured in late August and near the onion canopy (<2 m) throughout the season. However, 4% of T. tabaci adults captured on sticky cards were at altitudes ≥2 m, and T. tabaci were also captured on UAV-mounted traps. These data strongly suggest that long-distance dispersal occurs. More T. tabaci captured on sticky cards tested positive for IYSV in August (53.6%) than earlier in the season (2.3 to 21.5% in June and July, respectively), and 20 and 15% of T. tabaci captured on UAV-mounted traps tested positive for IYSV in 2012 and 2013, respectively. Our results indicate that T. tabaci adults, including viruliferous individuals, engage in long-distance dispersal late in the season and likely contribute to the spread of IYSV. © The Authors 2015. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

  6. The bottle gourd genome provides insights into Cucurbitaceae evolution and facilitates mapping of a Papaya ring-spot virus resistance locus.

    Science.gov (United States)

    Wu, Shan; Shamimuzzaman, Md; Sun, Honghe; Salse, Jerome; Sui, Xuelian; Wilder, Alan; Wu, Zujian; Levi, Amnon; Xu, Yong; Ling, Kai-Shu; Fei, Zhangjun

    2017-12-01

    Bottle gourd (Lagenaria siceraria) is an important vegetable crop as well as a rootstock for other cucurbit crops. In this study, we report a high-quality 313.4-Mb genome sequence of a bottle gourd inbred line, USVL1VR-Ls, with a scaffold N50 of 8.7 Mb and the longest of 19.0 Mb. About 98.3% of the assembled scaffolds are anchored to the 11 pseudomolecules. Our comparative genomic analysis identifies chromosome-level syntenic relationships between bottle gourd and other cucurbits, as well as lineage-specific gene family expansions in bottle gourd. We reconstructed the genome of the most recent common ancestor of Cucurbitaceae, which revealed that the ancestral Cucurbitaceae karyotypes consisted of 12 protochromosomes with 18 534 protogenes. The 12 protochromosomes are largely retained in the modern melon genome, while have undergone different degrees of shuffling events in other investigated cucurbit genomes. The 11 bottle gourd chromosomes derive from the ancestral Cucurbitaceae karyotypes followed by 19 chromosomal fissions and 20 fusions. The bottle gourd genome sequence has facilitated the mapping of a dominant monogenic locus, Prs, conferring Papaya ring-spot virus (PRSV) resistance in bottle gourd, to a 317.8-kb region on chromosome 1. We have developed a cleaved amplified polymorphic sequence (CAPS) marker tightly linked to the Prs locus and demonstrated its potential application in marker-assisted selection of PRSV resistance in bottle gourd. This study provides insights into the paleohistory of Cucurbitaceae genome evolution, and the high-quality genome sequence of bottle gourd provides a useful resource for plant comparative genomics studies and cucurbit improvement. © 2017 The Authors The Plant Journal © 2017 John Wiley & Sons Ltd.

  7. Virulence of white spot syndrome virus (WSSV) isolates may be correlated with the degree of replication in gills of Penaeus vannamei juveniles.

    Science.gov (United States)

    Rahman, M M; Corteel, M; Escobedo-Bonilla, C M; Wille, M; Alday-Sanz, V; Pensaert, M B; Sorgeloos, P; Nauwynck, H J

    2008-05-08

    A standardized inoculation model was used in 2 separate experiments to gauge the virulence of 3 white spot syndrome virus (WSSV) isolates from Thailand and Vietnam (WSSV Thai-1, WSSV Thai-2, and WSSV Viet) in Penaeus vannamei juveniles. Mortality patterns (Expt 1) were compared and WSSV-positive cells quantified (Expt 2) in tissues following intramuscular inoculation of shrimp with the most (WSSV Thai-1) and least (WSSV Viet) virulent isolates as determined by Expt 1. The results of Expt 1 demonstrated that mortalities began at 36 h post inoculation (hpi) for both Thai isolate groups and at 36 to 60 hpi for the Viet isolate group. Cumulative mortality reached 100% 96 to 240 h later in shrimp challenged with the WSSV Viet isolate compared to shrimp challenged with the Thai isolates. WSSV infection was verified in all groups by indirect immunofluorescence. In Expt 2, WSSV-infected cells were quantified by immunohistochemical analysis of both dead and time-course sampled shrimp. WSSV-positive cells were detected in tissues of Thai-1 inoculated dead and euthanized shrimp from 24 hpi onwards and from 36 hpi onwards in shrimp injected with the Viet isolate. Significantly more infected cells were found in tissues of dead shrimp inoculated with the Thai-1 than in Viet isolate-inoculated shrimp. In these experiments, substantial differences in virulence were demonstrated between the WSSV isolates. The Vietnamese isolate induced a more chronic disease and mortality pattern than was found for the Thai isolates, possibly because it infected fewer cells. This difference was most pronounced in gills.

  8. Hot spots

    National Research Council Canada - National Science Library

    Nia, Amir M; Gassanov, Natig; Er, Fikret

    2014-01-01

    ..., several reddened skin lesions were observed. The obvious ''hot spots'' were located on both sides in the groin and above the bladder, with extension to the genital region, compli- cating the ability to catheterize the patient (Figure 1). The rest of the body surface was not affected, and no infectious source for the skin lesions was evident. After suc...

  9. Word Spotting.

    Science.gov (United States)

    McQueen, James

    1996-01-01

    Summarizes the use of word-spotting in psycholinguistic research. Notes that listeners hear a list of nonsense words, some of which contain embedded real words, and they detect those embedded words, a task designed to study the segmentation of continuous speech. Describes the task and summarizes its advantages and disadvantages. (12 references)…

  10. Tissue Testing Can Spot Zika at Birth

    Science.gov (United States)

    ... news/fullstory_166828.html Tissue Testing Can Spot Zika at Birth: CDC Just 1 in 10 possible ... June 22, 2017 (HealthDay News) -- Exposure to the Zika virus in pregnancy can wreak havoc on babies, ...

  11. The N protein of Tomato spotted wilt virus (TSWV) is associated with the induction of programmed cell death (PCD) in Capsicum chinense plants, a hypersensitive host to TSWV infection.

    Science.gov (United States)

    Lovato, Fernanda Antinolfi; Inoue-Nagata, Alice Kazuko; Nagata, Tatsuya; de Avila, Antônio Carlos; Pereira, Luiz Alfredo Rodrigues; Resende, Renato Oliveira

    2008-11-01

    In sweet pepper, the Tsw gene, originally described in Capsicum chinense, has been widely used as an efficient gene for inducing a hypersensitivity response (HR) derived Tomato spotted wilt virus (TSWV) resistance. Since previously reported studies suggested that the TSWV-S RNA mutation(s) are associated with the breakdown of Tsw mediated TSWV resistance in peppers, the TSWV genes N (structural nucleocapsid protein) and NS(S) (non-structural silencing suppressor protein) were cloned into a Potato virus X (PVX)-based expression vector, and inoculated into the TSWV-resistant C. chinense genotype, PI 159236, to identify the Tsw-HR viral elicitor. Typical HR-like chlorotic and necrotic lesions followed by leaf abscission were observed only in C. chinense plants inoculated with the PVX-N construct. Cytopathological analyses of these plants identified fragmented genomic DNA, indicative of programmed cell death (PCD), in mesophyll cell nuclei surrounding PVX-N-induced necrotic lesions. The other constructs induced only PVX-like symptoms without HR-like lesions and there were no microscopic signs of PCD. The mechanism of TSWV N-gene HR induction is apparently species specific as the N gene of a related tospovirus, Tomato chlorotic spot virus, was not a HR elicitor and did not cause PCD in infected cells.

  12. Quarto ventrículo isolado: relato de dois casos

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    SILVA JOSÉ ALBERTO GONÇALVES DA

    1998-01-01

    Full Text Available São descritos dois casos de quarto ventrículo isolado sendo o primeiro decorrente de hemorragia cerebelar e o segundo de hidrocefalia congênita com múltiplas revisões de válvula e cisto de Dandy-Walker. O tratamento cirúrgico é revisado na literatura. A abordagem direta à fossa posterior acha-se indicada nos casos de cisto no interior do quarto ventrículo. Naqueles sem a presença de cisto, deve-se empregar sistema de drenagem do quarto ventrículo, independente da drenagem supratentorial.

  13. SUMO and SUMO-Conjugating Enzyme E2 UBC9 Are Involved in White Spot Syndrome Virus Infection in Fenneropenaeus chinensis.

    Science.gov (United States)

    Tang, Xiaoqian; Li, Wei; Xing, Jing; Sheng, Xiuzhen; Zhan, Wenbin

    2016-01-01

    In previous work, small ubiquitin-like modifier (SUMO) in hemocytes of Chinese shrimp Fenneropenaeus chinensis was found to be up-regulated post-white spot syndrome virus (WSSV) infection using proteomic approach. However, the role of SUMO in viral infection is still unclear. In the present work, full length cDNAs of SUMO (FcSUMO) and SUMO-conjugating enzyme E2 UBC9 (FcUBC9) were cloned from F. chinensis using rapid amplification of cDNA ends approach. The open reading frame (ORF) of FcSUMO encoded a 93 amino acids peptide with the predicted molecular weight (M.W) of 10.55 kDa, and the UBC9 ORF encoded a 160 amino acids peptide with the predicted M.W of 18.35 kDa. By quantitative real-time RT-PCR, higher mRNA transcription levels of FcSUMO and FcUBC9 were detected in hemocytes and ovary of F. chinensis, and the two genes were significantly up-regulated post WSSV infection. Subsequently, the recombinant proteins of FcSUMO and FcUBC9 were expressed in Escherichia coli BL21 (DE3), and employed as immunogens for the production of polyclonal antibody (PAb). Indirect immunofluorescence assay revealed that the FcSUMO and UBC9 proteins were mainly located in the hemocytes nuclei. By western blotting, a 13.5 kDa protein and a 18.7 kDa protein in hemocytes were recognized by the PAb against SUMO or UBC9 respectively. Furthermore, gene silencing of FcSUMO and FcUBC9 were performed using RNA interference, and the results showed that the number of WSSV copies and the viral gene expressions were inhibited by knockdown of either SUMO or UBC9, and the mortalities of shrimp were also reduced. These results indicated that FcSUMO and FcUBC9 played important roles in WSSV infection.

  14. SUMO and SUMO-Conjugating Enzyme E2 UBC9 Are Involved in White Spot Syndrome Virus Infection in Fenneropenaeus chinensis.

    Directory of Open Access Journals (Sweden)

    Xiaoqian Tang

    Full Text Available In previous work, small ubiquitin-like modifier (SUMO in hemocytes of Chinese shrimp Fenneropenaeus chinensis was found to be up-regulated post-white spot syndrome virus (WSSV infection using proteomic approach. However, the role of SUMO in viral infection is still unclear. In the present work, full length cDNAs of SUMO (FcSUMO and SUMO-conjugating enzyme E2 UBC9 (FcUBC9 were cloned from F. chinensis using rapid amplification of cDNA ends approach. The open reading frame (ORF of FcSUMO encoded a 93 amino acids peptide with the predicted molecular weight (M.W of 10.55 kDa, and the UBC9 ORF encoded a 160 amino acids peptide with the predicted M.W of 18.35 kDa. By quantitative real-time RT-PCR, higher mRNA transcription levels of FcSUMO and FcUBC9 were detected in hemocytes and ovary of F. chinensis, and the two genes were significantly up-regulated post WSSV infection. Subsequently, the recombinant proteins of FcSUMO and FcUBC9 were expressed in Escherichia coli BL21 (DE3, and employed as immunogens for the production of polyclonal antibody (PAb. Indirect immunofluorescence assay revealed that the FcSUMO and UBC9 proteins were mainly located in the hemocytes nuclei. By western blotting, a 13.5 kDa protein and a 18.7 kDa protein in hemocytes were recognized by the PAb against SUMO or UBC9 respectively. Furthermore, gene silencing of FcSUMO and FcUBC9 were performed using RNA interference, and the results showed that the number of WSSV copies and the viral gene expressions were inhibited by knockdown of either SUMO or UBC9, and the mortalities of shrimp were also reduced. These results indicated that FcSUMO and FcUBC9 played important roles in WSSV infection.

  15. Produção e sensibilidade de isolados brasileiros de Xanthomonas axonopodis pv. citri à bacteriocinas

    Directory of Open Access Journals (Sweden)

    Marcel Bonini

    2012-09-01

    Full Text Available Este trabalho teve por objetivo avaliar a produção e a sensibilidade de 48 isolados brasileiros de X. axonopodis pv. citri (Xac à bacteriocinas. Pelos resultados obtidos, nenhum isolado de Xac foi sensível às bacteriocinas produzidas pelos isolados bacterianos avaliados.

  16. Comportamento de híbridos de couve chinesa à isolados de Plasmodiophora brassicae

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    Daniel Dias Rosa

    2010-12-01

    Full Text Available Analisou-se o comportamento de cinco híbridos comerciais de couve chinesa perante a agressividade de 24 isolados de Plasmodiophora brassicae provenientes de diferentes regiões do estado de São Paulo. O ensaio foi conduzido em casa de vegetação e delineado em blocos ao acaso. Verificou-se que os cinco híbridos foram suscetíveis a todos os isolados de P.brassicae, sendo o hibrido 3 com comportamento mais resistente e o hibrido 2 com o comportamento mais suscetível. Dentre os isolados, verificou-se que os isolados de Piedade-SP foram os mais agressivos e os isolados Ribeirão Preto-SP foram os menos agressivos perante os cinco híbridos.

  17. Generation of an Infectious Clone of a New Korean Isolate of Apple chlorotic leaf spot virus Driven by Dual 35S and T7 Promoters in a Versatile Binary Vector

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    Ik-Hyun Kim

    2017-12-01

    Full Text Available The full-length sequence of a new isolate of Apple chlorotic leaf spot virus (ACLSV from Korea was divergent, but most closely related to the Japanese isolate A4, at 84% nucleotide identity. The full-length cDNA of the Korean isolate of ACLSV was cloned into a binary vector downstream of the bacteriophage T7 RNA promoter and the Cauliflower mosaic virus 35S promoter. Chenopodium quinoa was successfully infected using in vitro transcripts synthesized using the T7 promoter, detected at 20 days post inoculation (dpi, but did not produce obvious symptoms. Nicotiana occidentalis and C. quinoa were inoculated through agroinfiltration. At 32 dpi the infection rate was evaluated; no C. quinoa plants were infected by agroinfiltration, but infection of N. occidentalis was obtained.

  18. Efficacy of double-stranded RNA against white spot syndrome virus (WSSV non-structural (orf89, wsv191 and structural (vp28, vp26 genes in the Pacific white shrimp Litopenaeus vannamei

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    César M. Escobedo-Bonilla

    2015-04-01

    Full Text Available White spot syndrome virus (WSSV is a major pathogen in shrimp aquaculture. RNA interference (RNAi is a promising tool against viral infections. Previous works with RNAi showed different antiviral efficacies depending on the silenced gene. This work evaluated the antiviral efficacy of double-stranded (ds RNA against two non-structural (orf89, wsv191 WSSV genes compared to structural (vp26, vp28 genes to inhibit an experimental WSSV infection. Gene orf89 encodes a putative regulatory protein and gene white spot virus (wsv191 encodes a nonspecific nuclease; whereas genes vp26 and vp28 encode envelope proteins, respectively. Molecules of dsRNA against each of the WSSV genes were intramuscularly injected (4 μg per shrimp into a group of shrimp 48 h before a WSSV challenge. The highest antiviral activity occurred with dsRNA against orf89, vp28 and vp26 (cumulative mortalities 10%, 10% and 21%, respectively. In contrast, the least effective treatment was wsv191 dsRNA (cumulative mortality 83%. All dead animals were WSSV-positive by one-step PCR, whereas reverse-transcription PCR of all surviving shrimp confirmed inhibition of virus replication. This study showed that dsRNA against WSSV genes orf89, vp28 and vp26 were highly effective to inhibit virus replication and suggest an essential role in WSSV infection. Non-structural WSSV genes such as orf89 can be used as novel targets to design therapeutic RNAi molecules against WSSV infection.

  19. Mongolian spots

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    Divya Gupta

    2013-01-01

    Full Text Available Mongolian spots (MS are birthmarks that are present at birth and their most common location is sacrococcygeal or lumbar area. Lesions may be single or multiple and usually involve < 5% total body surface area. They are macular and round, oval or irregular in shape. The color varies from blue to greenish, gray, black or a combination of any of the above. The size varies from few to more than 20 centimetres. Pigmentation is most intense at the age of one year and gradually fades thereafter. It is rarely seen after the age of 6 years. Aberrant MS over occiput, temple, mandibular area, shoulders and limbs may be confused with other dermal melanocytoses and bruises secondary to child abuse, thus necessitating documentation at birth. Although regarded as benign, recent data suggest that MS may be associated with inborn errors of metabolism and neurocristopathies. Mongolian spots usually resolve by early childhood and hence no treatment is generally needed if they are located in the sacral area. However, sometimes it may be required for extrasacral lesions for cosmesis.

  20. Linfangioma cístico isolado do mediastino

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    Iury Andrade Melo

    2009-07-01

    Full Text Available Resumo: Doente de 59 anos, assintomática, com achado de uma tumoração ovariana na sua avaliação ginecológica de rotina e que durante os exames pré-operatórios encontrou acidentalmente um tumor mediastínico isolado, sendo então encaminhada para avaliação diagnóstica da lesão, que posteriormente mostrou ser um linfangioma cístico isolado do mediastino. O higroma cístico do mediastino é uma neoplasia benigna e extremamente infrequente, representando apenas 0,7 a 4,5% do total dos tumores mediastínicos e, dentre estes, apenas 1% é de localização exclusivamente mediastínica, sendo o diagnóstico definitivo possível apenas pelo exame anatomopatológico, e o tratamento recomendado consiste na ressecção cirúrgica completa. Os casos descritos são de relatos isolados ou séries com poucos doentes, e a sua predisposição ou sincronicidade a outros tumores é desconhecida, e, até onde conhecemos, não antes relatada. Abstract: A 59 years old female patient, asymptomatic, with the incidental finding of an ovarian tumor in her routine gynecological evaluation, and during the preoperative examinations it was incidentally found an isolated mediastinal tumor, and then routed to diagnostic evaluation of the lesion, which later proved to be a cystic lymphangioma. The cystic hygroma of the mediastinum is a benign tumor and very infrequent, representing only 0.7 to 4.5% of all mediastinal tumors, and of these, only 1% is exclusively mediastinal in location. The definitive diagnosis is only possible by pathological examination, and the recommended treatment consists of complete surgical resection. Cases are described in isolated reports or series with few patients, and their readiness or synchronicity with other tumors, unknown, and to the best of out knowledge, not reported yet. Palavras-chave: Linfangioma cístico, neoplasia mediastínica, cisto mediast

  1. Linfangioma cístico isolado do mediastino

    Directory of Open Access Journals (Sweden)

    Iury Andrade Melo

    2009-07-01

    Full Text Available Resumo: Doente de 59 anos, assintomática, com achado de uma tumoração ovariana na sua avaliação ginecológica de rotina e que durante os exames pré-operatórios encontrou acidentalmente um tumor mediastínico isolado, sendo então encaminhada para avaliação diagnóstica da lesão, que posteriormente mostrou ser um linfangioma cístico isolado do mediastino. O higroma cístico do mediastino é uma neoplasia benigna e extremamente infrequente, representando apenas 0,7 a 4,5% do total dos tumores mediastínicos e, dentre estes, apenas 1% é de localização exclusivamente mediastínica, sendo o diagnóstico definitivo possível apenas pelo exame anatomopatológico, e o tratamento recomendado consiste na ressecção cirúrgica completa. Os casos descritos são de relatos isolados ou séries com poucos doentes, e a sua predisposição ou sincronicidade a outros tumores é desconhecida, e, até onde conhecemos, não antes relatada.Rev Port Pneumol 2009; XV (4: 697-703 Abstract: A 59 years old female patient, asymptomatic, with the incidental finding of an ovarian tumor in her routine gynecological evaluation, and during the preoperative examinations it was incidentally found an isolated mediastinal tumor, and then routed to diagnostic evaluation of the lesion, which later proved to be a cystic lymphangioma. The cystic hygroma of the mediastinum is a benign tumor and very infrequent, representing only 0.7 to 4.5% of all mediastinal tumors, and of these, only 1% is exclusively mediastinal in location. The definitive diagnosis is only possible by pathological examination, and the recommended treatment consists of complete surgical resection. Cases are described in isolated reports or series with few patients, and their readiness or synchronicity with other tumors, unknown, and to the best of out knowledge, not reported yet.Rev Port Pneumol 2009; XV (4: 697-703 Palavras

  2. Development of a microarray for simultaneous detection and differentiation of different tospoviruses that are serologically related to Tomato spotted wilt virus

    OpenAIRE

    Liu, Lu-Yuan; Ye, He-Yi; Chen, Tsang-Hai; Chen, Tsung-Chi

    2017-01-01

    Background Tospoviruses, the plant-infecting genus in the family Bunyaviridae, are thrips borne and cause severe agricultural losses worldwide. Based on the serological relationships of the structural nucleocapsid protein (NP), the current tospoviruses are divided into six serogroups. The use of NP-antisera is convenient for virus detection, but it is insufficient to identify virus species grouped in a serogroup due to the serological cross-reaction. Alternatively, virus species can be identi...

  3. ANALISIS KEJADIAN SERANGAN WHITE SPOT SYNDROME VIRUS (WSSV DENGAN BEBERAPA PARAMETER KUALITAS AIR PADA BUDI DAYA UDANG WINDU MENGGUNAKAN SISTEM TANDON DAN PROBIOTIK

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    Gunarto Gunarto

    2006-08-01

    Full Text Available Munculnya serangan White Spot Syndrome Virus (WSSV pada udang yang dibudidayakan kemungkinan sebagai akibat menurunnya kualitas lingkungan tambak.  Data diperoleh dari penelitian budi daya udang windu yang dilakukan Balai Riset Perikanan Budidaya Air Payau, Maros menggunakan 8 unit tambak ukuran 500 m2. Tokolan udang windu PL-25 dengan padat tebar 10 dan 20 ekor/m2 ditebar dalam petak tambak tersebut serta penambahan probiotik setiap minggu sebanyak 1 mg/L  berlangsung selama pemeliharaan udang dan tanpa pemberian probiotik sebagai kontrol merupakan perlakuan yang diuji. Masing-masing perlakuan dengan dua ulangan. Setelah penebaran, beberapa petak terserang WSSV dan menyebabkan kematian total yaitu pada hari ke-27, 30, 41, dan 47. Serangan WSSV terus berlanjut selama pemeliharaan udang di tambak berlangsung. Pada petak menggunakan probiotik mempunyai  kecenderungan terserang WSSV lebih lambat daripada yang tidak menggunakan probiotik. Semakin tinggi padat tebar udang windu di tambak, maka semakin rentan terhadap serangan WSSV. Padat tebar 10 ekor/m2 menggunakan probiotik produksinya cenderung lebih baik daripada padat tebar 20 ekor/m2. Peningkatan populasi Vibrio sp., peningkatan konsentrasi nitrit dan tingginya populasi awal Vibrio sp. di air melebihi 103 cfu/mL dan di sedimen 104 cfu/g diduga erat kaitan dengan munculnya serangan WSSV pada udang yang dipelihara di tambak pada penelitian ini. The outbreak of WSSV infection on tiger shrimp culture was thought to be an impact of its pond environmental depletion. The data was obtained from the study of tiger shrimp culture conducted in ponds Research Station of RICA Maros using 8 unit of  brackishwater ponds compartment of 500 m2 each size. The PL-25 were stocked in the ponds at the density of 10 pieces and 20 pieces/m2 and on  the otherhand, ponds also were treated with 1 mg/L commercial probiotics applicated in every week during culture period and no probiotics application as control. Each

  4. Evaluation of peanut genotypes for resistance to Tomato spotted wilt virus by mechanical and thrips inoculation Avaliação de genótipos de amendoim em relação à resistência ao Tomato spotted wilt virus por meio de inoculação mecânica e por tripes

    Directory of Open Access Journals (Sweden)

    Luciana Cordeiro do Nascimento

    2006-06-01

    Full Text Available The objective of this work was to evaluate the reactions of three peanut breeding lines (IC-10, IC-34, and ICGV 86388 to Tomato spotted wilt virus (TSWV by mechanical and thrips inoculation, under greenhouse conditions, and compare them to the reactions of cultivars SunOleic, Georgia Green, and the breeding line C11-2-39. TSWV infection by mechanical inoculation was visually assessed using an index ranging from 0 (no symptoms to 4 (apical death. Enzyme-linked immunosorbent assay was used to confirm TSWV infection from both mechanical and thrips inoculations. IC-10, IC-34, ICGV 86388, and C11-2-39 were more resistant than the cultivars SunOleic and Georgia Green based on mechanical inoculation. Upon thrips inoculation only IC-34 and ICGV-86388 were infected by TSWV, as demonstrated by reverse transcription polymerase chain reaction (RT-PCR, although no symptoms of infection were observed. The peanut breeding lines IC-10, IC-34, and ICGV 86388 show higher level of resistance to TSWV than cultivar Georgia Green considered a standard for TSWV resistance.O objetivo deste trabalho foi avaliar as reações das linhagens de amendoim IC-10, IC-34 e ICGV 86388 quanto à resistência ao Tomato spotted wilt virus (TSWV, por meio de inoculação mecânica e por tripes, em casa de vegetação, e compará-las às reações das cultivares SunOleic, Georgia Green e da linhagem C11-2-39. A infecção de TSWV após inoculação mecânica foi visualmente avaliada, utilizando-se escala de notas que variam de zero (sem sintomas a quatro (morte apical. A análise pelo teste ELISA confirmou a infecção de TSWV em ambos os tipos de inoculação. As linhagens IC-10, IC-34, ICGV 86388 e C11-2-39 foram mais resistentes do que as cultivares SunOleic e Georgia Green, com base nos resultados da inoculação mecânica. Em relação à inoculação por tripes, apenas IC-34 e ICGV 86388 foram infectadas por TSWV, conforme demonstrado pela reação em cadeia da polimerase com

  5. Grafting on a Non-Transgenic Tolerant Tomato Variety Confers Resistance to the Infection of a Sw5- Breaking Strain of Tomato spotted wilt virus via RNA Silencing

    NARCIS (Netherlands)

    Spanò, R.; Mascia, T.; Kormelink, R.J.M.; Gallitelli, D.

    2015-01-01

    RNA silencing controls endogenous gene expression and drives defensive reactions against invasive nucleic acids like viruses. In plants, it has been demonstrated that RNA silencing can be transmitted through grafting between scions and silenced rootstocks to attenuate virus and viroid accumulation

  6. Tomato spotted wilt virus (TS WV, weeds and thrip vectors in the tomato (Solanum lycopersicum L. in the Andean region of Cundinamarca (Colombia

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    Everth E Ebratt R

    2013-04-01

    Full Text Available The presence and distribution of the TSWV, weeds and thrip vectors in major tomato producing areas in the Andean department of Cundinamarca (Oriente, Sumapaz and Ubate provinces were assessed with the DAS ELISA technique, evaluating the presence of the TSWV in tomato tissue, associated thrips and weeds. High incidences were observed in different provinces of the Andean department of Cundinamarca. The average viral incidence reached 23.3% in Sumapaz, 19.4% in Oriente and 4% in Ubate. The symptoms observed were: brown spots and concentric rings in the leaf area, stems and fruits; browning and spotting in the flower; and wilting in the leaves, stems and flowers. The thrip species with the highest presence were Frankliniella occidentalis, followed by Thrips palmi and Thrips tabaci. We determined the important role of weeds as inoculum sources and vector reservoirs for the species Emilia sonchifolia and Amaranthus dubius

  7. Desempenho agronômico de videiras com e sem sintomas de viroses, e comparação molecular de isolados virais

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    Monique Bezerra Nascimento

    2015-07-01

    Full Text Available Resumo: O objetivo deste trabalho foi avaliar os efeitos de viroses em videiras sintomáticas e assintomáticas sobre as variáveis agronômicas relacionadas ao vigor das plantas e à qualidade enológica da uva, e comparar os isolados virais obtidos nessas duas condições. Realizaram-se dois experimentos com quatro cultivares. Todas as plantas foram indexadas, por meio da reação em cadeia da polimerase via transcrição reversa (RT-PCR em tempo real, quanto à provável ocorrência dos seguintes vírus: Grapevine virus A (GVA, Grapevine virus B (GVB, Grapevine virus D (GVD, Grapevine leafroll-associated virus (GLRaV-1 ao -4, GLRaV-4 estirpe 5, Grapevine rupestris stem pitting-associated virus (GRSPaV e Grapevine fleck virus (GFkV. As variáveis avaliadas foram: número de gemas brotadas e não brotadas, número de ramos com ou sem cachos, número total de gemas, número de cachos, massa de cachos frescos, massa total de bagas, massa do engaço, número de bagas por cacho, massa média de baga, sólidos solúveis totais, acidez total titulável, pH, massa de ramos podados ou diâmetros do tronco do porta-enxerto e da copa. Os efeitos negativos foram mais pronunciados nas plantas com sintomas de viroses; no entanto, constatou-se frequentemente que plantas sem sintomas também estavam infectadas. A análise molecular de GRSPaV, GVA e GLRaV-2, isolados de plantas sintomáticas e assintomáticas, resultou em alta percentagem de identidade de nucleotídeos entre isolados homólogos.

  8. Genetic mapping of the Tsw locus for resistance to the Tospovirus Tomato spotted wilt virus in Capsicum spp. and its relationship to the Sw-5 gene for resistance to the same pathogen in tomato.

    Science.gov (United States)

    Jahn, M; Paran, I; Hoffmann, K; Radwanski, E R; Livingstone, K D; Grube, R C; Aftergoot, E; Lapidot, M; Moyer, J

    2000-06-01

    The Tsw gene conferring dominant resistance to the Tospovirus Tomato spotted wilt virus (TSWV) in Capsicum spp. has been tagged with a random amplified polymorphic DNA marker and mapped to the distal portion of chromosome 10. No mapped homologues of Sw-5, a phenotypically similar dominant TSWV resistance gene in tomato, map to this region in C. annuum, although a number of Sw-5 homologues are found at corresponding positions in pepper and tomato. The relationship between Tsw and Sw-5 was also examined through genetic studies of TSWV. The capacity of TSWV-A to overcome the Tsw gene in pepper and the Sw-5 gene in tomato maps to different TSWV genome segments. Therefore, despite phenotypic and genetic similarities of resistance in tomato and pepper, we infer that distinct viral gene products control the outcome of infection in plants carrying Sw-5 and Tsw, and that these loci do not appear to share a recent common evolutionary ancestor.

  9. Evidence for Lettuce big-vein associated virus as the causal agent of a syndrome of necrotic rings and spots in lettuce

    NARCIS (Netherlands)

    Verbeek, M.; Dullemans, A.M.; Bekkum, van P.J.; Vlugt, van der R.A.A.

    2013-01-01

    Lettuce big-vein associated virus (LBVaV, genus Varicosavirus) was shown to be responsible for characteristic necrotic symptoms observed in combination with big-vein symptoms in lettuce breeding lines when tested for their susceptibility to lettuce big-vein disease (BVD) using viruliferous Olpidium

  10. VIRUSES

    Indian Academy of Sciences (India)

    and-mouth disease in livestock was an infectious particle smaller than any bacteria. This was the first clue to the nature of viruses, genetic entities that lie somewhere in the gray area between living and non-living states.

  11. Caracterização de um vírus baciliforme isolado de Solanum violaefolium transmitido pelos ácaros Brevipalpus phoenicis e Brevipalpus obovatus (Acari: Tenuipalpidae Characterization of a bacilliform virus isolated from Solanum violaefolium transmitted by the tenuipalpid mites Brevipalpus phoenicis and Brevipalpus obovatus

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    Paulo de Tarso Oliveira Ferreira

    2007-09-01

    Full Text Available Solano-violeta (Solanum violaefolium é uma planta ornamental rasteira usada para cobrir solos de áreas sombreadas. Um vírus que induz manchas anelares nas folhas desta planta, tentativamente designado Solanum violaefolium ringspot virus - SvRSV, transmitido pelo ácaro Brevipalpus phoenicis (Acari: Tenuipalpidae foi encontrado em Piracicaba, SP. Trata-se de um vírus baciliforme que se assemelha a outros vírus do tipo citoplasmático transmitidos por Brevipalpus sp. Este trabalho teve como objetivo relatar propriedades biológicas e estabelecer uma caracterização molecular parcial do SvRSV. O vírus pode ser transmitido mecanicamente a várias outras espécies botânicas, causando lesões localizadas. Entre as espécies avaliadas, Datura stramonium mostrou-se a melhor hospedeira experimental. Observou-se também a manifestação de sintomas nestas plantas após infestação das mesmas por B. obovatus previamente alimentado em lesões de SvRSV, confirmando esta outra espécie de ácaro como vetor do vírus. Suas propriedades físicas in vitro foram: temperatura de inativação 40-45 ºC; ponto final de diluição 10-3-10-4; longevidade in vitro 12 dias. Em secções ultrafinas, as partículas do SvRSV mostraram-se levemente mais delgadas e mais longas que as de outros vírus do mesmo grupo. A partir do dsRNA do SvRSV foi construída uma biblioteca de cDNA e foram identificadas duas possíveis regiões codificadoras das proteínas de movimento e replicase viral. Baseado nestas regiões foram desenhados "primers" para amplificação do RNA do SvRSV por RT-PCR. Sondas baseadas nas seqüências obtidas hibridizaram com ss- e dsRNA de D. stramonium infectadas pelo vírus. Ensaios preliminares de RT-PCR e hibridização não resultaram em reação com o vírus da leprose dos citros, tipo citoplasmático (CiLV-C.Solanum violaefolium is an ornamental plant, with prostrate, trailing growth habit and is cultivated in shaded areas. A virus that causes

  12. Imunogenicidade de isolados de herpesvírus bovino 5 como candidatos à vacina Immunogenicity of bovine herpesvirus 5 isolates as vaccine candidates

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    Luiz Felipe Lourenço de Souza

    2009-02-01

    Full Text Available O herpesvírus bovino 5 (BoHV-5 é o agente da menigoencefalite herpética bovina. A doença neurológica, associada à infecção pelo BoHV-5, apresenta altas taxas de letalidade em bovinos jovens e está disseminada no Brasil. A prevenção das perdas causadas pela infecção pelo herpesvírus está baseada, principalmente, na imunização dos animais. Nesse sentido, foi delineada uma comparação entre isolados de BoHV-5, buscando selecionar o isolado mais antigênico para a formulação de vacinas. As formulações inativadas foram produzidas com os isolados ISO9898292, SV507, SV163, 1807 e EVI145 e administradas a cinco grupos de 10 ovelhas cada, que receberam duas doses vacinais por via intramuscular com intervalo de 21 dias. Foram realizadas coletas de sangue para análise de presença de anticorpos por soroneutralização e acompanhamento dos animais até o 63° dia após a primo-vacinação. Foram observados dois picos na curva de anticorpos, o primeiro no dia 14, após a vacinação, quando os títulos médios de anticorpos variaram entre 23,1 e 138,6. O segundo pico foi observado 14 dias após a revacinação, quando os títulos médios variaram entre 301,3 e 1017,5. No 42° dia após a revacinação, foi observada variação de título entre 82,4 e 305,9. A diferença entre as médias de títulos de anticorpos de cada grupo de animais sugere uma menor antigenicidade do isolado ISO9898292 em relação aos demais, demonstrando uma possível variação antigênica entre os isolados. Todos os isolados, com exceção do ISO9898292, mostraram-se imunogênicos para a indução de anticorpos.Herpesvirus bovine 5 (BoHV-5 is the agent of bovine herpetic menigoencephalitis. The neurological disease associated with the infection is highly lethal in young cattle and it is widespread in Brazil. Control of the clinical signs caused by herpesviruses is based mainly on the immunization of cattle. A comparative study was performed among Brazilian Bo

  13. Colonização do xilema de eucalipto por Ceratocystis spp. isolado de diferentes hospedeiros

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    Ana Carolina Firmino

    2015-06-01

    Full Text Available Ceratocystis sp. é um fungo que coloniza o xilema, agente causal de murcha e seca em diversas plantas lenhosas. O presente trabalho teve como objetivo acompanhar a colonização deste fungo na superfície de plantas de eucalipto (Eucalyptus sp.. Foram utilizados 5 isolados de Ceratocystis de diferentes hospedeiros (eucalipto, cacau, manga, teca e atemóia. Para isso, plantas de eucaliptos (clone 219 tiveram parte do seu sistema vascular exposto. Nesta região foi depositada uma suspensão contendo 106 esporos. Como testemunha, uma planta foi inoculada somente com água destilada autoclavada. Após a inoculação, estas plantas foram mantidas em câmara úmida a 25 oC no escuro. Parte da área inoculada foi coletada em intervalos de tempo pré-determinados (6, 12 e 24 horas, e fixadas em solução de "Karnovsky". As amostras foram preparadas e analisadas ao microscópio eletrônico de varredura. Todos os isolados foram capazes de germinar, penetrar e se desenvolver nos elementos de vasos de plantas de eucalipto no período de 6 horas. O isolado de cacau foi o que aparentemente teve o desenvolvimento mais lento dentro dos períodos estudados. Foi observada a germinação de ascósporos, clamidósporos e de conídios do tipo cilíndrico neste período. Doze horas após a inoculação ocorreu aumento da quantidade de micélio de todos os isolados testados. Nos casos dos isolados de manga e eucalipto foi possível observar a formação de novos clamidósporos. Vinte e quatro horas após a inoculação, com exceção do isolado de cacau, todos os outros isolados já apresentavam a formação de conidióforos cilíndricos. Este estudo comprova que isolados deste fungo, mesmo que provindos de outros hospedeiros, são capazes de se desenvolver no xilema do eucalipto.

  14. Advances in the processing of policromat images as diagnostic method to determine white spot syndrome virus in white shrimp (Litopenaeus vannamei)

    Science.gov (United States)

    Chavez-Sanchez, Cristina M.; Alvarez-Borrego, Josue; Montoya-Rodriguez, L.; Garcia-Gasca, A.; Fajer Avila, Emma J.; Pacheco-Marges, R.

    2004-10-01

    White spot syndrome (WSSV) is a viral disease which affects many crustacean species including commercial shrimps. Adequate, precise and quick methods to diagnose on time the presence of the disease in order to apply different strategies to avoid the dispersion and to reduce mortalities is necessary. Histopathology is an important diagnostic method. However, histopathology has the problem that requires time to prepare the histological slides and time to arrive to some diagnosis because this depend on the nature of the tissues, the pathogen(s) to find, the number of organisms, number of slides to analyze and the skill of the technician. This paper try to demonstrate the sensibility of one digital system of processing and recognition of images using color correlation with phase filters, to identify inclusion bodies of WSSV. Infected tissues were processed to obtain histological slides and to verify that the inclusion bodies observed were of WSV, in situ hybridization were carried out. The sensibility results of the recognition of the inclusion bodies of WSSV with the color correlation program was 86.1%. The highest percentage of recognition was in nervous system and tegument glands with 100%. The values in the stomach epithelium and heart tissue was 78.45% of recognition. Tissues with the lowest recognition values were lymphoid organ and hematopoietic tissue. It is necessary further studies to increase the sensibility and to obtain the specificity.

  15. Prevalence of the human immunodeficiency virus among patients with tuberculosis in Sierra Leone, established from dried blood spots on filter paper

    OpenAIRE

    Boillot, F.; Peeters, Martine; Kosia, A.; Delaporte, Eric

    1997-01-01

    Evaluation de l'utilisation sur le terrain de la technique des taches de sang séchées sur papier filtre pour la surveillance de la tuberculose associée au virus de l'immunodéficience humaine (VIH). La technique a été évaluée par comparaison avec le sérum chez 359 patients consentants enregistrés dans le registre de district de la tuberculose dans la capitale. Des tests répétés par ELISA utilisant des antigènes différents ont été effectués sur les éluats des papiers-filtres. Les sérums ont été...

  16. Ontogenetic changes in the expression of immune related genes in response to immunostimulants and resistance against white spot syndrome virus in Litopenaeus vannamei.

    Science.gov (United States)

    Rajkumar, T; Taju, G; Abdul Majeed, S; Sinwan Sajid, M; Santhosh Kumar, S; Sivakumar, S; Thamizhvanan, S; Vimal, S; Sahul Hameed, A S

    2017-11-01

    In recent years, researchers have focused on viral and plant immunostimulants which could have beneficial effects in disease prevention and control in shrimp culture. At present, the application of the recombinant VP28 protein (r-VP28) and herbal immunostimulant has been considered as a more effective approach to prevent white spot syndrome (WSS) by enhancing the immune response in shrimp. In the present study, expression of selected immune related genes in response to r-VP28 and herbal immunostimulant mix (HIM) were separately studied qualitatively and quantitatively by RT-PCR and real time PCR, respectively during ontogenetic development from nauplius to juvenile stage in Litopenaeus vannamei. The mRNA expression level of immune related genes such as anti-lipopolysaccharides (ALF), Lysozyme, cMnSOD, Crustin, Prophenoloxidase, Tumor necrosis factor receptor-associated factor 6 (TRAF6) and Haemocyanin were found to be up-regulated significantly in different ontogenetic development stages of shrimp fed with r-VP28 and HIM formulated diets. Relative percent survival (RPS) was determined in shrimp fed with immunostimulants formulated diets after oral challenge with WSSV. The survival of WSSV challenged shrimp was found to be higher in immunostimulants treated groups when compared to untreated group. The results of PCR, ELISA and real time PCR revealed the absence of WSSV in WSSV-challenged shrimp after 20 days of treatment with immunostimulants. Among these immunostimulants, HIM was found to be more effective when compared to r-VP28. After a survey of literature, we are of the opinion that this might be the first report on the expression of immune genes during ontogenetic development of L. vannamei in response to immunostimulants. Copyright © 2017 Elsevier Ltd. All rights reserved.

  17. Development of a rapid method for identifying carryover contamination of positive control DNA, using a chimeric positive control and restriction enzyme for the diagnosis of white spot syndrome virus by nested PCR.

    Science.gov (United States)

    Kim, Hyoung Jun; Kwon, Se Ryun

    2014-12-01

    Chimeric positive plasmids have been developed to minimize false-positive reactions caused by polymerase chain reaction (PCR) contamination. Here, we developed a rapid method for identifying false-positive results while detecting white spot syndrome virus (WSSV) by nested PCR, using chimeric positive plasmids. The results of PCRs using WSSV diagnostic primer sets showed PCR products of a similar size (WSSV 1st PCR product, 1,447 bp; WSSV 2nd PCR product, 941 bp) using WSSV chimeric plasmids or DNA from shrimp infected with WSSV. The PCR products were digested with DraI for 1 h at 37 °C. The digested chimeric DNA separated into two DNA bands; however, the WSSV-infected shrimp DNA did not separate. Thus, chimeric plasmid DNA may be used as positive control DNA instead of DNA from WSSV-infected shrimp, in order to prevent PCR contamination. Thus, the use of restriction enzyme digestion allowed us to rapidly distinguish between WSSV DNA and WSSV chimeric plasmid DNA.

  18. A reação de fixação do complemento na tipificação de vírus da gripe isolados no Rio de Janeiro The complement fixation test in the typefication of influenza viruses isolated in Rio de Janeiro

    Directory of Open Access Journals (Sweden)

    J. Guilherme Lacorte

    1976-01-01

    Full Text Available O autor estudou, pela reação de fixação do complemento, amostras de vírus da gripe isoladas no Rio de janeiro, durante a epidemia de 1973. Preparou imunesoros em hamsters pela inoculação do líquido alantóide de embriões de galinha infectados. o antígeno solúvel foi preparado com líquido obtido da mesma proveniência. As reações foram positivas, em grau variável, com as amostras clássicas PR8, FM1 e Ásia dos subtipos A0,A1 e A2 e as mais recentes A2/Hong Kong/68 e A2/England/72 e negativa com o anticorpo B/Mass/66. Para as duas variantes do subtipo A2, acima assinaladas e para as 7 amostras isoladas o comportamento foi praticamente o mesmo, não deixando de ser uma reação tipo específica se encararmos, também, as reações obtidas com as demais variantes do tipo A.The author studied by the complement fixation test the influenza virus strains isolated in Rio de Janeiro during the 1973 epidemic. He prepared immunesera in hamsters by the inoculation of the allantoic fluid from infected chick embryos with each of the 7 isolated strains and the standard strains. The soluble antigens were prepared with the allantoic fluid of infected chick embryos. The tests were identically positive with the A2/Hong Kong/68 and A2/England/72 antigens and negative with the B/Mass/66. The tets were type specific and the behaviour of the A2/Hong Kong/68 and the A2/England/72 and the 7 strains of the isolated viruses was almost the same. They fixed 3 or 4 units of complement. The variants PR8, FM1 and Asia fixed only 2 units of complement.

  19. Enhancement of superoxide dismutase and catalase activity in juvenile brown shrimp, Farfantepenaeus californiensis (Holmes, 1900, fed β-1.3 glucan vitamin E, and β-carotene and infected with white spot syndrome virus

    Directory of Open Access Journals (Sweden)

    Rosario Pacheco

    2011-11-01

    Full Text Available The effect of dietary β-Ο-glucan, vitamin E, and β-carotene supplements in juvenile brown shrimp, Farfantepenaeus californiensis, inoculated with white spot syndrome virus (WSSV was evaluated. Groups of 30 organisms (weighing 1 ± 0.5 g were cultured in 60 L fiberglass tanks and fed daily with β-1.3-glucan (0.1%, vitamin E (0.01%, and β-carotene (0.01% for 23 days; the specimens were then inoculated with WSSV. The antioxidant activity of the enzymes superoxide dismutase (SOD and catalase (CAT were determined in the hepatopancreas and muscle at 0, 1, 6, 12, 24, and 48 h after inoculation. Shrimp fed with β-1.3-glucan, vitamin E, and β-carotene significantly increased SOD activity in the hepatopancreas and muscle at 12 and 24 h post-infection, respectively. Shrimp fed with vitamin E and β-1.3-glucan registered an increment in SOD activity from 12 to 48 h post-infection. Shrimp fed with β-carotene increased SOD activity before infection with WSSV, and shrimp fed with β-1.3-glucan and vitamin E increased CAT activity, also before infection. The CAT activity response in shrimp muscle increased with respect to the control group for all treatments tested from 1 to 6 h after inoculation with WSSV. The highest antioxidant response was registered in shrimp fed with vitamin E. Juvenile shrimp fed with vitamin E and later inoculated with WSSV registered 100% mortality at 72 h, but shrimp fed with β-Ο-glucan and β-carotene showed greater resistance to WSSV, with mortality at 144 h post-infection. This study demonstrated the capacity of juvenile Farfantepenaeus californiensis fed β-Ο-glucan, vitamin E, or β-carotene to increase the antioxidant response before and after viral infection.

  20. Variabilidade genética em isolados de Curtobacterium flaccumfaciens Genetic variability in Curtobacterium flaccumfaciens isolates

    Directory of Open Access Journals (Sweden)

    Valmir Luiz de Souza

    2006-06-01

    Full Text Available A cultura do feijoeiro está sujeita à incidência de várias doenças que acarretam perdas significativas na produção, dentre as quais encontra-se a murcha-de-curtobacterium ou murcha bacteriana, causada por Curtobacterium flaccumfaciens pv. flaccumfaciens (Cff. Atualmente a murcha-de-curtobacterium tem se constituído em um novo problema para a cultura do feijoeiro em várias regiões brasileiras. A resistência genética tem sido o meio mais eficiente no controle da doença, porém a possibilidade da existência de variabilidade genética presente em isolados de Cff, pode ser uma conseqüência maléfica ao melhoramento visando a obtenção de cultivares de feijoeiro resistentes, especialmente na estabilidade e durabilidade da resistência. Neste sentido, o presente trabalho teve como objetivo o estudo da variabilidade genética de 26 isolados de Curtobacterium flaccumfaciens, 20 dos quais provenientes de feijoeiro (Cff, coletados em diferentes regiões do Brasil, quatro provenientes de coleções internacionais (Cff e dois endofíticos de citros (C. flaccumfaciens. Foram utilizados dois pares de oligonucleotídeos, CffFOR2-CffREV4 e CF4-CF5, avaliando-se na especificidade em reação de PCR, para a caracterização dos 26 isolados. No estudo da variabilidade genética, utilizou-se da técnica rep-PCR e os iniciadores REP, ERIC e BOX. A partir do padrão eletroforético gerado pela amplificação dessas seqüências repetitivas no DNA genômico dos 26 isolados bacterianos, foram realizadas análises pertinentes (UPGMA SM e obtenção de um dendograma. Considerando-se um índice de similaridade de 75%, os isolados foram distribuídos em quatro grupos distintos. Os isolados de Cff provenientes do Paraná e DF foram separados em grupos diferentes, enquanto que isolados endofíticos de citros não formaram um grupamento distinto dos de feijoeiro. Os isolados procedentes do Estados de São Paulo mostraram-se geneticamente heterogêneos, alguns

  1. ATIVIDADE BIOLÓGICA DE UM NOVO TRITERPENO ISOLADO DE Guarea cannata (MELIACEAE)

    OpenAIRE

    PEREIRA Júnior, Orlando L; WOLTER Filho, Wilson; WOLTER, Esther L. Aguiar; ZOGHBI, Maria das Graças B.; SIQUEIRA, Jane B. G.; PINHEIRO, Carlos C.S.

    1993-01-01

    Do cerne de Guarea carinata foi isolado como principal constituinte um triterpeno di-acetilado (I). do grupo dos protolimonóides, derivado do apo-tirucalol e que mostrou atividade no controle biológico das larvas de Urbanus acawoioa(WILLIAMS, R. C, 1926) (Lepidoptera Hesperiidac). From the bark of Guarea carinata was isolated as main constituent a diacetilated triterpen (I) of protolimonoids type, derived from apo-tirucalol, that showed antifeedant activity in biological control of Urbanus...

  2. Suscetibilidade a desinfetantes e perfil de resistência a antimicrobianos em isolados de Escherichia coli

    Directory of Open Access Journals (Sweden)

    Carina C. Krewer

    2012-11-01

    Full Text Available A colibacilose, causada por Escherichia coli, é a enfermidade entérica de maior impacto na produção de suínos, podendo levar à morte do animal. Esta bactéria possui grande capacidade de desenvolver resistência a múltiplos antimicrobianos e a desinfetantes. Desta forma, estudos que abordem mecanismos de resistência e perfil de amostras de campo tornam-se necessários. E. coli é amplamente utilizada como modelo de estudos que exploram a resistência intrínseca e extrínseca a multidrogas. Neste trabalho, buscou-se verificar o perfil de sensibilidade de 62 isolados de E. coli de suínos frente a três desinfetantes e a 13 antimicrobianos. Ainda, em 31 destes isolados foi pesquisada a presença de mecanismo de efluxo. Dos três desinfetantes avaliados, o cloreto de alquil dimetil benzil amônio+poliexietilenonilfenileter foi o que se mostrou mais eficaz (100%, seguido do glutaraldeído+cloreto de alquil dimetil benzil amônio (95,2% e do cloreto de alquil dimetil benzil amônio (88,8%. Dentre os antimicrobianos testados, observou-se maior resistência para a tetraciclina (62,2% e maior sensibilidade para o florfenicol (88,6%. A alta sensibilidade dos isolados frente aos desinfetantes pode estar relacionada à ausência de mecanismo de efluxo. O índice de resistência múltipla médio aos antimicrobianos foi de 0,52, o que demonstra um perfil multirresistente dos isolados, conduzindo para a necessidade do uso racional destas drogas em suinocultura.

  3. Mononucleosis spot test

    Science.gov (United States)

    Monospot test; Heterophile antibody test; Heterophile agglutination test; Paul-Bunnell test; Forssman antibody test ... The mononucleosis spot test is done when symptoms of mononucleosis are ... Fatigue Fever Large spleen (possibly) Sore throat Tender ...

  4. Spotted Seal Distribution Map

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — This dataset contains GIS layers that depict the known spatial distributions (i.e., ranges) and reported breeding areas of spotted seals (Phoca largha). It was...

  5. Concerning seed spots

    Science.gov (United States)

    J. A. Larsen; R. J. Smith

    1913-01-01

    In connection with the sowing of Yellow pine, White pine and Western larch on The Blackfeet National Forest during the sea sons of 1911 and 1912, seventeen and one-half acres were sowed directly in seed spots.

  6. Deep sequencing analysis of apple infecting viruses in Korea

    Science.gov (United States)

    Deep sequencing of viruses isolated from eight symptomatic apple trees in Korea has generated 52 contigs derived from five viruses: Apple chlorotic leaf spot virus (ACLSV), Apple stem grooving virus (ASGV), Apple stem pitting virus (ASPV), Apple green crinkle associated virus (AGCaV) and Apricot lat...

  7. Characterization of Xanthomonas spp. strains by bacteriocins Caracterização de isolados de Xanthomonas spp. por bacterocinas

    Directory of Open Access Journals (Sweden)

    Marcel Bonini

    2007-03-01

    Full Text Available Twenty-five strains of Xanthomonas axonopodis pv. citri and 14 strains of Xanthomonas spp. were tested for bacteriocin production. X. axonopodis pv. passiflorae strains were sensitive to the bacteriocins produced by the 25 X. axonopodis pv. citri strains evaluated in this study while strains of X. axonopodis pv. manihotis and X. campestris pv. campestris showed variable sensitivity. Only five of the 25 X. axonopodis pv. citri strains were not inhibited by the bacteriocins produced by the two X. axonopodis pv. passiflorae strains. The bacteriocins produced by the Xanthomonas axonopodis pv. citri (FDC-806 and X. axonopodis pv. passiflorae (Mar-2850 A strains were thermolabile, resistant to lysozyme and sensitive to DNAse. The bacteriocin produced by X. axonopodis pv. passiflorae was resistant to the action of proteinase K, trypsin and RNAse while the bacteriocin produced by X. axonopodis pv. citri was sensitive to these enzymes. The bacteriocins produced by X. axonopodis pv. passiflorae and X. axonopodis pv. citri were called passifloricin and citricin, respectively.Vinte e cinco isolados de Xanthomonas axonopodis pv. citri e 14 isolados de Xanthomonas spp. foram comparados a fim de verificar a capacidade de produção de bacteriocina e a sua sensibilidade. Isolados de X. axonopodis pv. passiflorae foram sensíveis às bacteriocinas produzidas por 25 isolados de X. axonopodis pv. citri avaliados e os isolados de X. axonopodis pv. manihotis e X. campestris pv. campestris apresentaram sensibilidade variável. Dos 25 isolados de X. axonopodis pv. citri apenas cinco não foram inibidos pelas bacteriocinas produzidas por dois isolados de X. axonopodis pv. passiflorae. As bacteriocinas produzidas pelos isolados de X. axonopodis pv. citri (FDC-806 e de X. axonopodis pv. passiflorae (Mar-2850 A foram termolábeis e resistentes à lisozima e sensíveis a DNAse. A bacteriocina produzida pelo isolado de X. axonopodis pv. passiflorae foi resistente à a

  8. New thrips-transmitted plant viruses in Florida crops

    Science.gov (United States)

    The thrips-transmitted tospoviruses Tomato spotted wilt virus, Groundnut ringspot virus and Tomato chlorotic spot virus are present in south Florida. All three species cause economically significant disease in vegetable and ornamental crops, and may also be problematic in peanut. Control of both t...

  9. Chlorotic spots on Clerodendrum, a disease caused by a nuclear type of Brevipalpus (Acari:Tenuipalpidae transmitted virus Mancha clorótica do Clerodendrum, uma enfermidade causada por um vírus do tipo nuclear, transmitido pelo ácaro Brevipalpus phoenicis (Acari:Tenuipalpidae

    Directory of Open Access Journals (Sweden)

    Elliot Watanabe Kitajima

    2008-02-01

    Full Text Available Chlorotic spots have been observed in plants of Clerodendrum x speciosum growing in residential gardens and parks in Piracicaba, SP, Brazil. Thin sections of diseased tissues revealed characteristic cytopathic effects of the nuclear type of the Brevipalpus (Acari: Tenuipalpidae mite-transmitted viruses (BTrV. Brevipalpus mites, identified as B. phoenicis, infesting symptomatic C. x speciosum plants transmitted the pathogen to healthy C. x speciosum and to C. thomsonae, Gomphrena globosa, Hibiscus cannabinus, H. coccineus, H. schizopetalus, Salvia leucantha, Spathiphyllum wallasi and Tetragonia expansa causing chlorotic spots on their leaves. Mechanical inoculation using leaf extracts from infected C. x speciosum resulted in chlorotic spots on inoculated C. x speciosum, Chenopodium quinoa, C. amaranticolor, G. globosa, H. cannabinus, H. coccineus and T. expansa leaves. C. amaranticolor and C. quinoa kept at 28 - 30°C became systemically infected. The same cytopathic effects caused by the nuclear type of BTrV were seen in tissues from all infected test plants by electron microscopy. The virus was purified from systemically infected leaves of C. amaranticolor and C. quinoa. A polyclonal antiserum obtained from an immunized rabbit presented a strong reaction with the homologous antigen in ELISA tests. The results suggest that this chlorotic spot disease of C. x speciosum is caused by a new species of the nuclear type of BTrV, tentatively named Clerodendrum chlorotic spot virus (ClCSV.Manchas cloróticas e necróticas foram observadas em folhas de várias plantas de coração-sangrento (Clerodendrum x speciosum cultivadas em parques e jardins em Piracicaba, SP, associadas à infestação pelo ácaro tenuipalpídeo Brevipalpus phoenicis. Exames preliminares de secções de tecido das manchas cloróticas ao microscópio eletrônico revelaram a ocorrência de efeitos citopáticos característicos dos induzidos pelos vírus do tipo nuclear, transmitido

  10. Fitoconstituintes isolados da fração em diclorometano das folhas de Vernonia tweediana Baker

    Directory of Open Access Journals (Sweden)

    Ricardo Basso Zanon

    Full Text Available Fracionamento cromatográfico da fração em diclorometano obtida do extrato etanólico das folhas de Vernonia tweediana Baker (Asteraceae conduziu ao isolamento de α-amirina, β-amirina, lupeol, β-sitosterol, estigmasterol e espinasterol. As estruturas foram identificadas através de técnicas espectroscópicas usuais, além da comparação com dados relatados na literatura. Os compostos isolados são relatados pela primeira vez para a espécie V. tweediana.

  11. SpotADAPT

    DEFF Research Database (Denmark)

    Kaulakiene, Dalia; Thomsen, Christian; Pedersen, Torben Bach

    2015-01-01

    Having constantly increasing amounts of data, the analysis of it is often entrusted for a MapReduce framework. The execution of an analytical workload can be cheapened by adopting cloud computing resources, and in particular by using spot instances (cheap, fluctuating price instances) offered...

  12. Degradação de xenobióticos por fungos filamentosos isolados de areia fenólica

    OpenAIRE

    J. H. Silva; Monteiro,R.T.R.

    2000-01-01

    Microrganismos foram isolados de areia fenólica resultante de atividades metalúrgicas, utilizando meio mínimo para fungos e pentaclorofenol (PCF) como única fonte de carbono. Após quatro repiques sucessivos em intervalos de 15 dias de incubação, as culturas foram plaqueadas em meio de Martin. Três gêneros de fungos foram isolados e identificados como Acremonium sp., Paecilomyces sp. e Penicillium sp. Estes foram testados para degradar os corantes índigo e RBBR (Azul Brilhante de Remazol - R) ...

  13. Morfologia de conídios e patogenicidade de isolados de Exserohilum turcicum da Argentina e do Brasil em milho

    Directory of Open Access Journals (Sweden)

    Roberto Luis De Rossi

    2015-03-01

    Full Text Available A cultura do milho é uma das principais commodities da Argentina e do Brasil. Seu rendimento é reduzido por agentes nocivos, sendo um deles o fungo Exserohilum turcicum (Et agente causal da helmintosporiose do milho.O presente trabalho teve como objetivo comparar as características morfológicas dos conídios e a patogenicidade de dez isolados de Et obtidos da Argentina e do Brasil. Cinco isolados monospóricos de cada país foram cultivados em meio lactose caseina hidrolizada ágar (LCHA durante 15 dias a 25+2ºC e na ausência de luz. Prepararam-se lâminas microscópicas para a mensuração de 200 conidios de cada isolado. Na média os conídios mediram 10-25 x 30-135 μm, apresentando 2-8 septos. Detectaram-se diferenças estatísticas (p = 0,05 entre os dez isolados de Et para todas as variáveis medidas (comprimento, largura e número de septos. Apesar da variação, as características mensuradas coincidiram com as registradas na literatura. Na comprovação da patogenicidade, plantas do híbrido de milho Pioneer P1630H, de conhecida suscetibilidade a Et, foram inoculadas com a deposição de 0,5 mL de uma suspensão de conídios de Et na concentração de 5x104 conídios/mL, no cartucho, quando atingiram a quarta folha expandida.Os dez isolados produziram, após os 15 dias da inoculação, uma média de 2,5 lesões/folha; de 39,7 x 3,4 mm de comprimento e largura respetivamente; e 4% de severidade estimada. Não se observou diferenças significativas na patogenicidade dos dez isolados estudados para nenhum dos critérios patométricos. Houve diferenças morfológicas entre os isolados da Argentina e do Brasil, porém não quanto à patogenicidade. Confirma-se que os isolados utilizados neste trabalho pertencem à espécie Et.

  14. Poisson Spot with Magnetic Levitation

    Science.gov (United States)

    Hoover, Matthew; Everhart, Michael; D'Arruda, Jose

    2010-01-01

    In this paper we describe a unique method for obtaining the famous Poisson spot without adding obstacles to the light path, which could interfere with the effect. A Poisson spot is the interference effect from parallel rays of light diffracting around a solid spherical object, creating a bright spot in the center of the shadow.

  15. Resistência antimicrobiana de coliformes isolados de leite humano ordenhado

    Directory of Open Access Journals (Sweden)

    Novak Franz Reis

    2001-01-01

    Full Text Available A veiculação de microrganismos potencialmente patogênicos, resistentes aos antibióticos, por meio de leite humano ordenhado, pode ser um fator de risco potencial. Este estudo teve como objetivo contribuir para um melhor conhecimento dos coliformes isolados de leite humano ordenhado (LHO e de seu perfil de resistência a antimicrobianos. Seguiu-se um delineamento inteiramente casualizado, utilizando 837 amostras de LHO, nas quais 71 (8,48% estavam contaminadas com coliformes totais, cuja contagem em nenhuma das amostras ultrapassava 1,0x10³NMP/ml. A maioria dos microrganismos isolados (91,6% pertencia a apenas duas espécies, Enterobacter cloacae e Klebsiella pneumoniae. As 71 cepas de coliformes apresentavam resistência a pelo menos um dos antimicrobianos testados. Conclui-se que o LHO coletado e armazenado sob condições higiênico-sanitárias insatisfatórias, pode apresentar coliformes, que desgastam os fatores de proteção, reduzem o valor nutricional e a qualidade do produto.

  16. Resistência antimicrobiana de coliformes isolados de leite humano ordenhado

    Directory of Open Access Journals (Sweden)

    Franz Reis Novak

    2001-06-01

    Full Text Available A veiculação de microrganismos potencialmente patogênicos, resistentes aos antibióticos, por meio de leite humano ordenhado, pode ser um fator de risco potencial. Este estudo teve como objetivo contribuir para um melhor conhecimento dos coliformes isolados de leite humano ordenhado (LHO e de seu perfil de resistência a antimicrobianos. Seguiu-se um delineamento inteiramente casualizado, utilizando 837 amostras de LHO, nas quais 71 (8,48% estavam contaminadas com coliformes totais, cuja contagem em nenhuma das amostras ultrapassava 1,0x10³NMP/ml. A maioria dos microrganismos isolados (91,6% pertencia a apenas duas espécies, Enterobacter cloacae e Klebsiella pneumoniae. As 71 cepas de coliformes apresentavam resistência a pelo menos um dos antimicrobianos testados. Conclui-se que o LHO coletado e armazenado sob condições higiênico-sanitárias insatisfatórias, pode apresentar coliformes, que desgastam os fatores de proteção, reduzem o valor nutricional e a qualidade do produto.

  17. Sequence diversity in the coat protein gene of Lettuce big-vein associated virus and Mirafiori lettuce big-vein virus infecting lettuce in Brazil Variabilidade genética na porção codificadora para a proteína capsidial do Lettuce big-vein associated virus e Mirafiori lettuce big-vein virus provenientes de alface no Brasil

    Directory of Open Access Journals (Sweden)

    Márcio Martinello Sanches

    2008-06-01

    Full Text Available Lettuce big vein associated virus (LBVaV and Mirafiori lettuce big vein virus (MLBVV have been found in mixed infection in Brazil causing the lettuce big vein disease. Analysis of part of the coat protein (CP gene of Brazilian isolates of LBVaV collected from lettuce, showed at least 93% amino acid sequence identity with other LBVaV isolates. Genetic diversity among MLBVV CP sequences was higher when compared to LBVaV CP sequences, with amino acid sequence identity ranging between 91% to 100%. Brazilian isolates of MLBVV belong to subgroup A, with one RsaI restriction site on the coat protein gene. There is no indication for a possible geografical origin for the Brazilian isolates of LBVaV and MLBVV.Lettuce big vein associated virus (LBVaV e Mirafiori lettuce big vein virus (MLBVV têm sido encontrados em infecções mistas no Brasil, causando a doença conhecida como engrossamento das nervuras da alface. Análise de parte do gene da proteína capsidial (CP de isolados brasileiros de LBVaV coletados em alface, indicou que estes possuem identidade superior a 93% com isolados coletados em diferentes regiões geográficas. A diversidade genética entre a CP de isolados de MLBVV de alface foi maior comparada às sequências da CP de LBVaV, com a identidade de aminoácidos variando entre 91 a 100%. Os isolados brasileiros de MLBVV pertencem ao subgrupo A, com um único sítio de restrição RsaI no gene da proteína capsidial. Não há indicação para uma provável origem geográfica dos isolados brasileiros de MLBVV e LBVaV.

  18. Blind spot crashes.

    OpenAIRE

    2009-01-01

    Crashes involving lorries turning right and cyclists going straight ahead usually have very serious consequences for the cyclist. The cyclist, who has right of way, is often overlooked by the lorry driver. For his part, the cyclist is often unaware that the lorry driver has not seen him or that the driver wants to turn right. Despite a variety of measures, this type of blind spot crashes continues to occur. Each year they still cause approximately ten fatalities. This number could be reduced ...

  19. Bioprospecção de isolados de Bacillus spp. como potenciais promotores de crescimento de Eucalyptus urograndis

    Directory of Open Access Journals (Sweden)

    Ana Ligia de Lima Moreira

    2013-10-01

    Full Text Available Objetivou-se neste trabalho avaliar características bioquímicas de interesse agronômico e correlação com a promoção de crescimento de plantas em isolados de Bacillus sp. originários da rizosfera de eucalipto. O trabalho foi conduzido em laboratório e casa de vegetação. A partir do isolamento de bactérias da rizosfera de plantas, oriundas de diferentes municípios da região oeste paulista, conseguiu-se 127 isolados de Bacillus spp. Foram realizados testes bioquímicos para caracterização dos isolados bacterianos quanto ao antagonismo a fungos fitopatogênicos, produção de auxinas, produção de amônia e atividade enzimática. Na etapa final foi avaliado o potencial dos isolados, caracterizados previamente em condições de laboratório, para promoção de crescimento de plantas, utilizando-se a inoculação das bactérias em mudas de eucalipto e cultivo das plantas em casa de vegetação durante 90 dias. Avaliou-se variáveis de crescimento do eucalipto objetivando-se selecionar os melhores isolados e também correlacionar as diferentes variáveis analisadas no trabalho. O protocolo de bioprospecção de Bacillus sp. na rizosfera foi válido para se encontrar rizobactérias promissoras no aumento do crescimento do eucalipto. Foram selecionados cinco isolados como promissores para ação no crescimento de eucalipto. O potencial antagônico a fungos fitopatogênicos e produção de amônia apresentados pelos isolados de rizobactérias foi útil na fase inicial de seleção de rizobactérias promotoras do crescimento de eucalipto, pois apresentou correlação significativa com o crescimento das plantas.

  20. The Spotting Distribution of Wildfires

    Directory of Open Access Journals (Sweden)

    Jonathan Martin

    2016-06-01

    Full Text Available In wildfire science, spotting refers to non-local creation of new fires, due to downwind ignition of brands launched from a primary fire. Spotting is often mentioned as being one of the most difficult problems for wildfire management, because of its unpredictable nature. Since spotting is a stochastic process, it makes sense to talk about a probability distribution for spotting, which we call the spotting distribution. Given a location ahead of the fire front, we would like to know how likely is it to observe a spot fire at that location in the next few minutes. The aim of this paper is to introduce a detailed procedure to find the spotting distribution. Most prior modelling has focused on the maximum spotting distance, or on physical subprocesses. We will use mathematical modelling, which is based on detailed physical processes, to derive a spotting distribution. We discuss the use and measurement of this spotting distribution in fire spread, fire management and fire breaching. The appendix of this paper contains a comprehensive review of the relevant underlying physical sub-processes of fire plumes, launching fire brands, wind transport, falling and terminal velocity, combustion during transport, and ignition upon landing.

  1. ATIVIDADE ANTIMICROBIANA DO EXTRATO DE PRÓPOLIS FRENTE A ISOLADOS DE STAPHYLOCOCCUS COAGULASE POSITIVA E MALASSEZIA PACHYDERMATIS DE OTITE CANINA

    OpenAIRE

    Rosemari Laura Cardoso

    2009-01-01

    O objetivo do presente estudo foi avaliar o potencial antimicrobiano do extrato de própolis, substância natural conhecida pelas suas propriedades terapêuticas, frente a 67 isolados microbianos da otite canina. A Concentração Bactericida Mínima (CBM), do extrato de própolis, frente a isolados de Staphylococcus coagulase positiva e a Concentração Fungicida Mínima (CFM) para isolados de Malassezia pachydermatis foram determinadas utilizando-se a técnica de microdiluição em caldo. A CBM foi...

  2. Feijoeiro manteiga, planta-teste para os vírus de vira-cabeça e da necrose branca do fumo A bean variety useful as a local-lesion test plant for tomato spotted wilt and Brazilian tobacco streak viruses

    Directory of Open Access Journals (Sweden)

    A. S. Costa

    1957-01-01

    mistura de búfer e sulfito de sódio deu o maior aumento.Out of 200 bean varieties tested, plants of the var. Manteiga were the most sensitive to tomato spotted wilt (TSW and Brazilian tobacco streak (BTS viruses. The inoculated plants developed local chlorotic spots adequate for counts within 3-6 days after inoculation with the TSW virus; pin point or rings in 2-4 days following inoculation with the BTS virus. Bean plants with the primary leaves 2/3 expanded or slightly older gave better results for the TSW virus, whereas they were more sensitive to BTS virus when 2/3 expanded or slightly younger. The TSW virus did not become systemic in the bean plants. Most strains of the BTS virus also did not become systemic in the inoculated plants. A yellow strain of this virus usually did. Sodium sulfite at 0.01 M added to the infected tissues during extraction of the inoculum increased the number of lesions formed in the bean leaves inoculated with the TSW virus; the increase was greater when extraction was made in presence of phosphate buffer at pH 7 and at the concentration of 0.1 M. A mixture of buffer and sulfite did not cause a greater increase than buffer alone. Por the BTS virus the addition of sodium sulfite at 0.01 M during extraction gave a large increase in the number of lesions; buffer alone caused only a small increase, but a mixture of buffer and sodium sulfite gave the highest increase. Phosphate buffer at 0.05 M and with pH 7 or 8, added prior to extraction, gave a higher number of lesions in case of both viruses than the same concentration of buffer at pH 5 or 6. The use of butter at concentrations ranging from 0.1 to 0.0125 M were tried and did not cause great differences in the number of local lesions, but the best ones seemed to be 0.05 or 0.025 M. Four concentrations of sodium sulfite, 0.1, 0.05, 0.025, and 0.0125 M were compared as diluents for the same inocula in presence of phosphate buffer. The two lowest ones gave the highest number of lesions for both

  3. Caracterização de isolados de Xanthomonas axonopodis pv. manihotis = Characterization of isolates of Xanthomonas axonopodis pv. manihotis

    Directory of Open Access Journals (Sweden)

    Roberto Luis Portz

    2006-07-01

    Full Text Available A bacteriose (Xanthomonas axonopodis pv. manihotis e a doenca de maior importancia economica na cultura da mandioca. Para estudar a variabilidade genetica desta bacteria no Oeste do Parana, foram realizados levantamentos em Entre Rios do Oeste, Marechal Candido Rondon, Mercedes, Missal, Nova Santa Rosa e Pato Bragado. Os isolados foram caracterizados em relacao a atividade de amilase, ƒ¿ e ƒÀ-esterase e agressividade. Dos 61 materiais vegetais coletados, obtiveram-se 19 isolados da bacteria, com maior incidencia para variedades de mesa em relacao aqueles para industria. Manivas provenientes de Pato Bragado, Entre Rios do Oeste e Mercedes apresentaram incidencias de 10, 27 e 10%, respectivamente, valores inferiores aos de Marechal Candido Rondon (50% e Nova Santa Rosa (58%. Os isolados foram agrupados em cinco, seis e 12 grupos em relacao a capacidade amilolitica, agressividade e isoenzimas de esterase, respectivamente. Nao houve relacao entre atividade de amilase e agressividade. Isolados de Marechal Candido Rondon foram mais agressivos que os provenientes das outras regioes. O agrupamento com base em esterase permitiu verificar que isolados provenientes de Entre Rios do Oeste, Nova Santa Rosa e Mercedes apresentaram alto grau de similaridade. Estes resultados indicam haver diferenciacaoentre os isolados da bacteria presentes nos municipios amostrados.The bacterial blight (Xanthomonas axonopodis pv. manihotis is the most important disease of cassava. To study the genetic variability of pathogen in the West of Parana, a research was carried out at Entre Rios do Oeste, Marechal Candido Rondon, Mercedes, Missal, Nova Santa Rosa and Pato Bragado. The isolates were characterized to amylase activity, ƒ¿ and ƒÀ-esterase and aggressiveness. From 61 collected materials, were obtained 19 bacterial isolates, with larger incidence for varieties of human consume than those for industry. Stems from Pato Bragado, Entre Rios do Oeste and Mercedes

  4. Compatibility and ectomycorrhiza formation among Pisolithus Isolates and Eucalyptus spp Compatibilidade e formação de Ectomicorrizas entre Isolados de Pisolithus e Eucalyptus spp

    Directory of Open Access Journals (Sweden)

    Olinto Liparini Pereira

    2005-06-01

    Full Text Available Twenty-nine isolates of the ectomycorrhiza fungus Pisolithus sp. from different geographical and host origins were tested for their ability to form ectomycorrhizae on Eucalyptus grandis and E. urophylla seedlings under greenhouse conditions. The ectomycorrhiza-forming capacity of isolates varied greatly from one eucalypt species to the other. All isolates from Eucalyptus, nine from Pinus spp. and two isolates from unknown hosts formed mycorrhizae with E. grandis and E. urophylla. Root colonization rates varied from 0 to 5.2 % for all Pinus isolates and those from unknown hosts. Colonization rates for these isolates were lower than those observed for Eucalyptus isolates (0.8 to 89.4 %. Three isolates from unknown hosts formed mycorrhizae with neither Eucalyptus species. The main characteristic for distinguishing Pinus from Eucalyptus isolates was mantle color. These data corroborate previous results obtained in our laboratory indicating that the isolates tested represent at least two distinct different species within the genus Pisolithus.Vinte e nove isolados do fungo ectomicorrízico Pisolithus sp., de diferentes regiões geográficas e hospedeiros, foram testados quanto à capacidade de formar ectomicorrizas em plântulas de Eucalyptus grandis e E. urophylla sob condições de casa de vegetação. Os isolados apresentaram grande variação na capacidade de formar ectomicorrizas com ambas as espécies de eucalipto. Todos os isolados originalmente obtidos de Eucalyptus, 9 originalmente obtidos de Pinus spp. e dois isolados de hospedeiros desconhecidos formaram micorrizas com E. grandis e E. urophylla. A taxa de colonização radicular dos isolados originalmente obtidos de Pinus e dos isolados de hospedeiros desconhecidos variou de 0 a 5,2 %. A taxa de colonização para esses isolados foi menor do que as obtidas para os isolados originalmente obtidos de Eucalyptus (0,8 a 89,4 %. Três isolados obtidos de hospedeiros desconhecidos não formaram

  5. Avaliação da atividade antibacteriana do óleo essencial de Lippia origanoides frente à Staphylococcus sp. isolados de alimentos de origem animal

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    M.R.A. Queiroz

    2014-01-01

    Full Text Available Objetivou-se no trabalho avaliar a ação antibacteriana do óleo essencial de Lippia origanoides frente à isolados de Staphylococcus sp. de alimentos de origem animal. Para tanto, realizou-se análise química da composição do óleo, teste de sensibilidade das bactérias frente a dez antibióticos de uso terapêutico e ao óleo essencial de Lippia origanoides, além da determinação da concentração inibitória mínima (CIM e da concentração bactericida mínima (CBM. A análise cromatográfica do óleo apresentou o timol como composto majoritário (48,70%, além de pequena quantidade de carvacrol (1,14%. No teste de sensibilidade frente aos antibióticos, 75% dos isolados apresentaram resistência a, no mínimo, três antibióticos. Em relação ao óleo essencial, os isolados de Staphylococcus sp oriundos de leite bovino mostraram-se mais resistentes e os isolados de carne ovina apresentaram-se mais sensíveis. A CIM foi maior para os Staphylococcus sp. isolados de leite bovino e leite ovino (60µL/mL. Enquanto que para os isolados de carcaça de ovinos e queijo, a CIM foi de 15µL/mL e 30µL/mL respectivamente. A CBM, consequentemente, foi maior para os isolados de leite bovino e leite ovino, sendo de 240µL/mL e 120µL/mL respectivamente. O óleo essencial de Lippia origanoides apresenta atividade antimicrobiana sobre Staphylococcus sp. isolados de alimentos.

  6. Perfil antigênico do vírus da raiva isolado de diferentes espécies de morcegos não hematófagos da Região de Presidente Prudente, Estado de São Paulo Antigen profile of rabies virus isolated from different species of non-hematophagous bats in the region of Presidente Prudente, State of São Paulo

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    Avelino Albas

    2009-02-01

    Full Text Available O Laboratório de Virologia Clínica e Molecular do Instituto de Ciências Biomédicas da Universidade de São Paulo, utilizando-se da técnica de anticorpos monoclonais, tipificou 18 amostras de vírus rábico provenientes de morcegos não hematófagos de várias espécies provenientes da Região de Presidente Prudente, SP, Brasil. Destas amostras, 15 (82,3% foram definidas como variante 3 (compatível com amostras isoladas de morcegos Desmodus rotundus e 3 (16,7% como variante 4 (compatível com amostras isoladas de morcegos Tadarida brasiliensis.Using the monoclonal antibody technique, the Clinical and Molecular Virology Laboratory of the Institute of Biomedical Sciences of the University of São Paulo typed 18 rabies virus samples from non-hematophagous bats of several species from the region of Presidente Prudente, SP, Brazil. Among these samples, 15 (82.3% were defined as variant 3 (compatible with samples isolated from Desmodus rotundus bats and three (16.7% as variant 4 (compatible with samples isolated from Tadarida brasiliensis bats.

  7. Black-spot poison ivy.

    Science.gov (United States)

    Schram, Sarah E; Willey, Andrea; Lee, Peter K; Bohjanen, Kimberly A; Warshaw, Erin M

    2008-01-01

    In black-spot poison ivy dermatitis, a black lacquerlike substance forms on the skin when poison ivy resin is exposed to air. Although the Toxicodendron group of plants is estimated to be the most common cause of allergic contact dermatitis in the United States, black-spot poison ivy dermatitis is relatively rare.

  8. Identification of potential hot spots in the carboxy-terminal part of the Epstein-Barr virus (EBV) BNLF-1 gene in both malignant and benign EBV-associated diseases

    DEFF Research Database (Denmark)

    Sandvej, K; Peh, S C; Andresen, B S

    1994-01-01

    In this study, we have sequenced the C-terminal part of the Epstein-Barr virus (EBV)-BNLF-1 gene encoding for the latent membrane protein-1 from tissues of EBV-positive Danish Hodgkin's disease (HD) and of Danish and Malaysian peripheral T-cell lymphomas (PTLs) and from tonsils of Danish infectious...

  9. Progressive outer retinal necrosis presenting as cherry red spot.

    Science.gov (United States)

    Yiu, Glenn; Young, Lucy H

    2012-10-01

    To report a case of progressive outer retinal necrosis (PORN) presenting as a cherry red spot. Case report. A 53-year-old woman with recently diagnosed HIV and varicella-zoster virus (VZV) aseptic meningitis developed rapid sequential vision loss in both eyes over 2 months. Her exam showed a "cherry red spot" in both maculae with peripheral atrophy and pigmentary changes, consistent with PORN. Due to her late presentation and the rapid progression of her condition, she quickly developed end-stage vision loss in both eyes. PORN should be considered within the differential diagnosis of a "cherry red spot." Immune-deficient patients with a history of herpetic infection who present with visual loss warrant prompt ophthalmological evaluation.

  10. Seleção e caracterização molecular de isolados de Bacillus thuringiensis para o controle de Spodoptera spp.

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    Kelly Christiane Constanski

    2015-08-01

    Full Text Available Resumo:O objetivo deste trabalho foi selecionar e caracterizar molecularmente isolados de Bacillus thuringiensistóxicos a Spodoptera eridaniaeS. frugiperda. Trinta e quatro isolados foram submetidos ao bioensaio, dos quais três foram selecionados e usados para a estimativa da CL50. Os isolados selecionados não diferiram da linhagem padrão HD-1. Na caracterização molecular, identificou-se a presença dos genes cry1 e cry2, nos isolados BR37 e BR94, e dos genes cry4A, cry4B, cry10, cry11 e cyt1 no isolado BR58, o que confirmou o perfil proteico obtido de 130, 70 e 65 kDa. Foram identificados cristais bipiramidais e esféricos. O isolado BR58, apesar de não conter os genes relacionados à toxicidade a Lepidoptera, causa mortalidade em ambas as espécies

  11. Caracterização de novos isolados de Bacillus thuringiensis para o controle de importantes insetos-praga da agricultura

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    Emeline Boni Campanini

    2012-01-01

    Full Text Available A bactéria Bacillus thuringiensis Berliner produz um corpo de inclusão paraesporal (cristal de natureza proteica, formado durante a esporulação, que atua de forma eficiente no controle de insetos-praga de culturas economicamente importantes. Esse cristal é constituído de proteínas Cry, que são codificadas pelos genes cry; um isolado pode ser caracterizado pelo conteúdo de genes cry que apresenta. Visando caracterizar novos isolados no combate de insetos-praga pertencentes às ordens Lepidoptera e Coleoptera, 76 isolados bacterianos foram analisados molecularmente e tiveram seu potencial de controle avaliado por meio de bioensaios com larvas de Spodoptera frugiperda (J.E. Smith, Sphenophorus levis Vaurie e Tenebrio molitor Linnaeus. As análises moleculares indicaram 11 isolados (14,5% da coleção, contendo genes lepidóptero-específicos e 17 (22,37% com genes coleóptero-específicos. As análises de patogenicidade revelaram dois isolados com alto potencial de controle para lagartas de S. frugiperda, um para larvas de S. levis e seis prejudiciais ao desenvolvimento das larvas de T. molitor. Esses isolados de B. thuringiensis podem ser promissores no controle biológico das referidas pragas.

  12. Hot Spot Cosmic Accelerators

    Science.gov (United States)

    2002-11-01

    length of more than 3 million light-years, or no less than one-and-a-half times the distance from the Milky Way to the Andromeda galaxy, this structure is indeed gigantic. The region where the jets collide with the intergalactic medium are known as " hot spots ". Superposing the intensity contours of the radio emission from the southern "hot spot" on a near-infrared J-band (wavelength 1.25 µm) VLT ISAAC image ("b") shows three distinct emitting areas; they are even better visible on the I-band (0.9 µm) FORS1 image ("c"). This emission is obviously associated with the shock front visible on the radio image. This is one of the first times it has been possible to obtain an optical/near-IR image of synchrotron emission from such an intergalactic shock and, thanks to the sensitivity and image sharpness of the VLT, the most detailed view of its kind so far . The central area (with the strongest emission) is where the plasma jet from the galaxy centre hits the intergalactic medium. The light from the two other "knots", some 10 - 15,000 light-years away from the central "hot spot", is also interpreted as synchrotron emission. However, in view of the large distance, the astronomers are convinced that it must be caused by electrons accelerated in secondary processes at those sites . The new images thus confirm that electrons are being continuously accelerated in these "knots" - hence called "cosmic accelerators" - far from the galaxy and the main jets, and in nearly empty space. The exact physical circumstances of this effect are not well known and will be the subject of further investigations. The present VLT-images of the "hot spots" near 3C 445 may not have the same public appeal as some of those beautiful images that have been produced by the same instruments during the past years. But they are not less valuable - their unusual importance is of a different kind, as they now herald the advent of fundamentally new insights into the mysteries of this class of remote and active

  13. Crescimento e esporulação de isolados de Verticillium lecanii sob diferentes condições nutricionais

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    Barbosa Claudio Camargo

    2002-01-01

    Full Text Available O objetivo deste trabalho foi determinar as melhores condições de crescimento e esporulação de dois isolados de Verticillium lecanii (Zimm. Viégas, cultivados em vários meios de cultura, fontes de C, fontes orgânicas e inorgânicas de N e relações C:N. Após 20 dias de cultivo, BDA e Meio Completo mostraram ser os melhores meios de cultura, e a lactose e o amido foram as fontes de C que mais favoreceram o crescimento dos isolados JAB 02 e JAB 45, respectivamente; baixa esporulação foi apresentada por JAB 02, enquanto JAB 45 obteve boa esporulação em glicose e maltose. JAB 02 desenvolveu melhor quando a fonte orgânica de N era casitona; JAB 45 obteve bom desenvolvimento em presença de casitona e peptona, mas houve pouca esporulação em presença de triptona e caseína hidrolisada. As fontes inorgânicas de N que favoreceram o melhor desenvolvimento dos isolados foram (NH42HPO4 e NaNO3, porém, JAB 02 apresentou baixa esporulação. Entre as relações C:N analisadas, JAB 02 obteve melhor crescimento na relação 60:1 e JAB 45 nas relações 5:1, 10:1 e 20:1; baixa esporulação foi obtida por JAB 02 nas relações testadas, mas não se verificou efeito da relação C:N do meio sobre a esporulação dos isolados.

  14. Degradação de xenobióticos por fungos filamentosos isolados de areia fenólica

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    J. H. Silva

    2000-09-01

    Full Text Available Microrganismos foram isolados de areia fenólica resultante de atividades metalúrgicas, utilizando meio mínimo para fungos e pentaclorofenol (PCF como única fonte de carbono. Após quatro repiques sucessivos em intervalos de 15 dias de incubação, as culturas foram plaqueadas em meio de Martin. Três gêneros de fungos foram isolados e identificados como Acremonium sp., Paecilomyces sp. e Penicillium sp. Estes foram testados para degradar os corantes índigo e RBBR (Azul Brilhante de Remazol - R e o organoclorado PCF. A descoloração do índigo foi de 99%, para Paecilomyces e Penicillium, e de 74%, para Acremonium, e a de RBBR foi de 16%, para Penicillium; 14%, para Acremonium, e 5%, para Paecilomyces. Usando azul de bromotimol como indicador de degradação de PCF, foram obtidos 24% de descoloração para Acremonium; 22%, para Penicillium, e 17%, para Paecilomyces Utilizando cromatografia gasosa, detectou-se degradação de PCF de 69%, para Penicillium; 65%, para Paecilomyces, e 40% para Acremonium, respectivamente. Os resultados mostraram que foi possível isolar microrganismos de uma areia de fundição, altamente contaminada com fenóis, e os fungos isolados foram capazes de degradar PCF e outros xenobióticos testados.

  15. Genetic variability of Brazilian isolates of Alternaria alternata detected by AFLP and RAPD techniques Variabilidade genética de isolados Brasileiros de Alternaria alternata por meio de marcadores moleculares de AFLP e RAPD

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    Francisco Dini-Andreote

    2009-09-01

    Full Text Available The Alternaria brown spot (ABS is a disease caused in tangerine plants and its hybrids by the fungus Alternaria alternata f. sp. citri which has been found in Brazil since 2001. Due to the recent occurrence in Brazilian orchards, the epidemiology and genetic variability of this pathogen is still an issue to be addressed. Here it is presented a survey about the genetic variability of this fungus by the characterization of twenty four pathogenic isolates of A. alternata f. sp. citri from citrus plants and four endophytic isolates from mango (one Alternaria tenuissima and three Alternaria arborescens. The application of two molecular markers Random Amplified Polymorphic DNA (RAPD and Amplified Fragment Length Polymorphism (AFLP had revealed the isolates clustering in distinct groups when fingerprintings were analyzed by Principal Components Analysis (PCA. Despite the better assessment of the genetic variability through the AFLP, significant modifications in clusters components were not observed, and only slight shifts in the positioning of isolates LRS 39/3 and 25M were observed in PCA plots. Furthermore, in both analyses, only the isolates from lemon plants revealed to be clustered, differently from the absence of clustering for other hosts or plant tissues. Summarizing, both RAPD and AFLP analyses were both efficient to detect the genetic variability within the population of the pathogenic fungus Alternaria spp., supplying information on the genetic variability of this species as a basis for further studies aiming the disease control.A mancha marrom ou mancha de Alternaria é uma doença causada pelo fungo Alternaria alternata f. sp. citri, encontrada no Brasil desde 2001 em plantas de tangerina e seus híbridos. Por se tratar de uma doença recente no Brasil, a epidemiologia e variabilidade genética deste patógeno compõem importantes pontos a serem estudados. Este trabalho teve como objetivo avaliar a variabilidade genética deste pat

  16. Análise da diversidade genética de isolados de Xanthomonas axonopodis pv. malvacearum do algodoeiro Analysis of genetic diversity among the isolates of Xanthomonas axonopolis pv. malvacearum of cotton

    Directory of Open Access Journals (Sweden)

    Maria Paula Nunes

    2009-06-01

    Full Text Available A mancha angular do algodoeiro, causada por Xanthomonas axonopodis pv. malvacearum (Xam, é uma doença de importância econômica e pode causar perdas apreciáveis no rendimento. A doença pode ser controlada por resistência varietal desde que haja conhecimento sobre a variabilidade das populações do patógeno. A variabilidade genética e a estabilidade patogênica entre os isolados deste patógeno não foram suficientemente estudadas, principalmente considerando a introdução de novas cultivares e a expansão da cultura. O objetivo do presente estudo foi avaliar a variabilidade genética entre os 61 isolados de Xam, provenientes de diversas cultivares e regiões do Brasil, através de ensaios moleculares. Análises de ERIC e REP-PCR demonstraram dois grupos distintos de Xam associados a região geográfica de origem. Não foram observadas diferenças nos perfis dos isolados através de PCR-RFLP da região 16S-23S rDNA. A região espaçadora 16S-23S rDNA de três linhagens de Xam foi analisada através de clonagem e sequenciamento e seis diferenças nas seqüências foram encontradas. A técnica de RAPD revelou um maior nível de polimorfismo, distinguindo 6 grupos de Xam a 85% de similaridade. Os resultados indicam a existência de variabilidade muito restrita entre os isolados analisados.Angular leaf spot or black arm of cotton, caused by Xanthomonas axonopodis pv. malvacearum (Xam, is a disease of economic importance. The disease can be controlled by using resistant cultivars if the knowledge regarding the genetic variability of the pathogen population is available. The genetic variability and the pathogen stability among the isolates of this pathogen have not been sufficiently studied, especially considering the introduction of new cultivars and the expansion of areas under cotton cultivation. The objective of the present study was to identify genetic variability among 61 isolates of Xam collected from different cultivars and geographic

  17. Suscetibilidade antimicrobiana de Staphylococcus spp. isolados de cães com pioderma superficial

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    Ana Paula da Silva

    2014-04-01

    Full Text Available Foram obtidas 154 amostras de lesões cutâneas de cães com pioderma superficial atendidos no Serviço de Dermatologia Veterinária do Hospital Veterinário Universitário (HVU da Universidade Federal de Santa Maria (UFSM, com o objetivo de determinar a suscetibilidade antimicrobiana de isolados de Staphylococcus spp. e avaliar a presença de multirresistência. Após isolamento e identificação, as cepas foram submetidas ao teste de sensibilidade aos antimicrobianos, cujos resultados evidenciaram menores percentuais de resistência à associação amoxicilina e ácido clavulânico (1,9%, cefadroxil (1,9%, cefalexina (1,9% e vancomicina (0,6%. Os maiores percentuais de resistência foram frente à amoxicilina (60,4% e penicilina G (60,4%. A multirresistência foi detectada em 23,4% e a resistência à meticilina em 5,8% das amostras. Pode-se concluir que os isolados de Staphylococcus spp. apresentam elevada suscetibilidade aos antimicrobianos comumente utilizados no tratamento dos piodermas superficiais em cães atendidos no HVU-UFSM, como a cefalexina e a amoxicilina associada ao ácido clavulânico, confirmando a eleição desses fármacos para o tratamento de cães com esta afecção. A suscetibilidade diminuída das cepas frente às fluoroquinolonas, também recomendadas pela literatura para o tratamento de pioderma, permite sugerir que estes fármacos não devem mais ser considerados na seleção empírica. A identificação de Staphylococcus spp. multirresistentes na população canina estudada justifica análises bacteriológicas periódicas e regionais de lesões cutâneas de cães com pioderma superficial, a fim de minimizar a seleção de bactérias resistentes, possíveis falhas terapêuticas e também motiva a antimicrobianoterapia prudente.

  18. Caracterização parcial de um Tymovirus isolado de tomateiros

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    Isabel Cristina Batista

    2012-01-01

    Full Text Available Um Tymovirus isolado de sementes de tomateiro (Solanum lycopersicum 'Paronset' com sintomas de necrose, denominado TyToRS08, foi caracterizado quanto ao círculo de hospedeiras, propriedades físico-químicas, sorológicas e moleculares. O vírus foi transmitido mecanicamente a partir de folhas, frutos e sementes infectadas. A gama de hospedeiras incluiu espécies de Amaranthaceae, Chenopodiaceae e Solanaceae, induzindo sintomas local e sistêmicos na maior parte das espécies, exceto para Nicotiana sylvestris, N. tabacum 'Samsun', 'White Burley' e 'Xanthi' que não manifestaram sintomas. Em tomateiros 'Alambra', 'Thomas', 'Diva', 'Romana' e 'Graziela', o vírus induziu sintomas de mosaico e necrose branca. A presença do vírus foi detectada, nas sementes infectadas de todas as cultivares avaliadas, porém não se notou em plântulas após a germinação destas sementes. Foi obtida uma preparação purificada com concentração de 1,04 mg mL-1, empregada na produção de um antissoro policlonal que reagiu em PTA-ELISA com um título de 1/32768. Utilizou-se um par de oligonucleotídeos degenerados, desenhados para anelar na ORF 1 de espécies de Tymovirus, que permitiu a amplificação por RT-PCR de fragmentos com cerca de 700 pb. Após o alinhamento e a análise das sequências, verificou-se que o isolado TyToRS08 possuía valores abaixo de 70% de identidade com as espécies de Tymovirus, indicando que pode se tratar de uma espécie distinta das descritas no gênero, com potencial epidemiológico, pela sua estabilidade e disseminação por sementes de tomate.

  19. Isolados de Dicyma pulvinata em estromas de Microcyclus ulei em seringueira Dicyma pulvinata isolates colonizing Microcyclus ulei stromata in rubber

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    Sueli Corrêa Marques de Mello

    2006-02-01

    Full Text Available Dicyma pulvinata é um eficiente agente de biocontrole de Microcyclus ulei, causador do mal-das-folhas da seringueira. O objetivo deste trabalho foi obter isolados com grande potencial antagônico. Em levantamentos realizados em 14 municípios produtores de borracha, localizados nos Estados do Acre, Amazonas, Bahia, Mato Grosso, Pará e Rondônia, obtiveram-se 52 isolados de D. pulvinata. O fungo antagônico, isolado diretamente em meio de cultura de batata-dextrose-ágar (BDA, foi identificado com base na morfologia dos conídios e conidióforos e preservado pelos métodos da liofilização, congelamento à temperatura de -80ºC e criopreservação em nitrogênio líquido, a fim de manter as características morfológicas e patogênicas dos isolados. O efeito antagônico foi testado por meio de inoculações de D. pulvinata em lesões de Fusicladium macrosporum induzidas em plantas de seringueira previamente infectadas. Todos os isolados foram incorporados à coleção de fungos da Embrapa Recursos Genéticos e Biotecnologia.Dicyma pulvinata is an efficient biocontrol agent of Microcyclus ulei, causal agent of South American leaf blight. The objective of this work was to obtain strongly antagonistic isolates. A survey was carried out in rubber plantations located in 14 municipalities in the Sates of Acre, Amazonas, Bahia, Mato Grosso, Pará and Roraima, Brazil, obtaining 52 isolates of D. pulvinata. The antagonistic fungus was isolated on potato-dextrose-agar (PDA plates and identified based on morphological characteristics of conidiophores and conidia, and colony appearance. The antagonic effect was tested by inoculating D. pulvinata on Fusicladium macrosporum lesions induced on rubber plants, previously. In order to maintain the morphological and pathogenic traits, fungus isolates were preserved by lyophilization, deep freeze (-80ºC and cryogeny. All isolates were introduced into Embrapa Recursos Genéticos e Biotecnologia fungus collection.

  20. Cianobactérias e algas reduzem os sintomas causados por Tobacco vosaic virus (tmv em plantas de fumo

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    André B. Beltrame

    2011-06-01

    Full Text Available As algas e as cianobactérias produzem uma grande diversidade de compostos com atividade biológica direta sobre microrganismos ou agem como ativadores de mecanismos de resistência em plantas. Em vista disso, foi investigada a manifestação dos sintomas causados pelo Tobacco mosaic virus (TMV em plantas de fumo previamente tratadas com cianobactérias ou algas. Quando as folhas plantas de fumo foram tratadas dois dias antes da inoculação, foi verificado que suspensões de células dos isolados de cianobactérias 004/02, 008/02, Anabaena sp. e Nostoc sp. 61; e do isolado de alga 061/02, bem como as preparações do conteúdo intracelular do isolado 004/02 (4 C e do filtrado do meio de cultivo do isolado 061/02 (61 M apresentaram efeito na redução do número de lesões locais provocadas por TMV em folhas de plantas fumo, cultivar TNN. Além disso, foi observado que os isolados Anabaena sp., Nostoc sp. 21 (cianobactéria, Nostoc sp. 61 e 090/02 (alga mostraram efeito direto sobre o vírus semi-purificado. Em vista disso, pode-se sugerir que os isolados estudados sintetizam compostos que agem diretamente sobre o TMV e/ou ativam o mecanismo de defesa de plantas contra fitopatógenos.

  1. Artefatos cumarínicos isolados de Polygala paniculata L. (Polygalaceae

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    M. G Pizzolatti

    Full Text Available A partir do fracionamento em coluna de gel de sílica do extrato clorofórmico de Polygala paniculata (Polygalaceae, foram isolados 7-metóxi-8-(1',2'-epóxi-3'-metil-3'-butenil-cumarina (1 e dois artefatos cumarínicos (2a-2b, formados a partir da abertura do anel epóxido de 1 durante o processo de isolamento. O tratamento de 1 com EtOH/SiO2, sob agitação e à temperatura ambiente durante 24 horas, resultou na formação dos respectivos artefatos: 7-metóxi-8-(1'-hidróxi-2-etóxi-3'-metil-3'-butenil-cumarina (2a e 7-metóxi-8-(1'-etóxi-2-hidróxi-3'-metil-3'-butenil-cumarina (2b. As estruturas químicas desses compostos foram determinadas através da análise de seus dados espectrais, incluindo RMN bidimensional.

  2. Caracterização da agressividade e atividade enzimática de isolados de Colletotrichum spp. associados à antracnose do abacate

    Directory of Open Access Journals (Sweden)

    Hugo José Tozze Júnior

    Full Text Available RESUMO A antracnose é a principal doença pós-colheita do abacate. O presente trabalho teve como objetivos caracterizar a agressividade e a atividade enzimática de 30 isolados de Colletotrichum spp. de abacate. Abacates ‘Fuerte’ foram inoculados com disco de BDA contendo estruturas de cada isolado e após sete dias mensuraram-se os diâmetros das lesões. Dois grupos de isolados foram distinguidos quanto à agressividade pelo teste de teste de Scott-Knott (p=0,05, um com lesões entre 34,0 e 38,2 mm e outro com lesões entre 38,7 e 44,0 mm de diâmetro. Para caracterização enzimática, avaliaram-se as áreas dos halos formados em substratos específicos para detecção de amilase, celulase, lacase, lipase, pectato liase e protease e a atividade da enzima catalase. Os isolados não produziram amilase suficiente para formação de halo em meio de cultura. Todos os isolados produziram catalase, pectato liase e protease, enquanto 46% produziram lacase e 97% tiveram as atividades da celulase e lipase detectadas. Distinguiram-se dois grupos de isolados para as enzimas catalase e pectato liase, cinco grupos para a celulase, seis grupos para a lacase e sete grupos para as enzimas lipase e protease, pelo teste de teste de Scott-Knott (p=0,05. Não houve relação entre as atividades enzimáticas e a agressividade dos isolados.

  3. Biological stability of a strain of Cowpea severe mosaic virus over 20 years Estabilidade biológica de uma estirpe do Cowpea severe mosaic virus ao longo de 20 anos

    Directory of Open Access Journals (Sweden)

    José Albersio Araujo Lima

    2012-03-01

    Full Text Available Cowpea (Vigna unguiculata is an important crop of the traditional agriculture system in the Northeast of Brazil. It can be infected by more than 20 virus species and Cowpea severe mosaic virus (CPSMV is one of the most important pathogens that naturally infect cowpea in Brazil. Several CPSMV isolates were obtained and characterized in the Plant Virus Laboratory at the Federal University of Ceará: CPSMV-CE - the first characterized isolate of the virus obtained from cowpea in the State of Ceará; CPSMV-AL - isolated from cowpea in Alagoas; CPSMV-PE - isolated from cowpea in Pernambuco; CPSMV-PR - obtained from soybean (Glycine max in Paraná and CPSMV-CROT - isolated from Crotalaria paulinea, in Maranhão. An isolate of CPSMV with the property to infect the cv. Macaibo, a cowpea cultivar immune to most of CPSMV isolates was also biologically and serologically characterized as a new strain of the virus (CPSMV-MC. The CPSMV-MC was isolated in January 1990 and has been evaluated over 20 years by host range studies and maintenance in vivo by periodical mechanical inoculations in cowpea. The results of this periodical evaluation revealed that the biological integrity and the serological properties of CPSMV-MC were preserved over 20 years, indicating that the genetic preservation of a virus strain could occur over the years. Molecular studies involving part of the coat protein (CP gene of CPSMV-MC and five other Brazilian CPSMV isolates indicated a high degree of conservation, with 92-100% nucleotide sequence identity among the isolates.O feijão-caupi (Vigna unguiculata é uma cultura do sistema tradicional do Nordeste do Brasil, que pode ser infetada por mais de 20 espécies de vírus, sendo o vírus do mosaico severo do caupi (Cowpea severe mosaic virus, CPSMV um dos mais importantes patógenos que infeta naturalmente essa leguminosa no Brasil. Vários isolados do CPSMV foram obtidos e caracterizados no Laboratório de Virologia Vegetal da UFC

  4. Caracterização de isolados de Diplodia pinea da região Sul do Brasil por meio da compatibilidade micelial e de marcadores RAPD

    Directory of Open Access Journals (Sweden)

    Paula Rachel Rabelo Corrêa Corrêa

    2016-03-01

    Full Text Available RESUMO O objetivo do trabalho foi caracterizar quatro isolados de Diplodiapinea da região Sul do Brasil e estimar sua variabilidade genética, baseados na compatibilidade vegetativa e marcadores RAPD. Na compatibilidade vegetativa, os isolados foram pareados em placas de Petri com meio BDA e formaram linhas escuras quando incompatíveis e linhas claras cotonosas quando incompatíveis. Para a caracterização molecular dos isolados, a extração de DNA foi realizada em amostras obtidas de micélio cultivado em meio BDA dos isolados originais e os monospóricos derivados. O DNA das amostras foi avaliado em reação de polimerase em cadeia (PCR, utilizando onze sequências diferentes de primers RAPD inespecíficos. O agrupamento dos morfotipos foi realizado pelo método UPGMA e coeficiente de similaridade de Jaccard. Somente um marcador RAPD mostrou polimorfismo, indicando que os isolados apresentam pequena divergência genética, porém suficiente para indicar mais de morfotipo presente.

  5. Performance of an Early Infant Diagnostic Test, AmpliSens DNA-HIV-FRT, Using Dried Blood Spots Collected from Children Born to Human Immunodeficiency Virus-Infected Mothers in Ukraine.

    Science.gov (United States)

    Chang, Joy; Tarasova, Tetyana; Shanmugam, Vedapuri; Azarskova, Marianna; Nguyen, Shon; Hurlston, Mackenzie; Sabatier, Jennifer; Zhang, Guoqing; Osmanov, Saladin; Ellenberger, Dennis; Yang, Chunfu; Vitek, Charles; Liulchuk, Maria; Nizova, Natalya

    2015-12-01

    An accurate accessible test for early infant diagnosis (EID) is crucial for identifying HIV-infected infants and linking them to treatment. To improve EID services in Ukraine, dried blood spot (DBS) samples obtained from 237 HIV-exposed children (≤18 months of age) in six regions in Ukraine in 2012 to 2013 were tested with the AmpliSens DNA-HIV-FRT assay, the Roche COBAS AmpliPrep/COBAS TaqMan (CAP/CTM) HIV-1 Qual test, and the Abbott RealTime HIV-1 Qualitative assay. In comparison with the paired whole-blood results generated from AmpliSens testing at the oblast HIV reference laboratories in Ukraine, the sensitivity was 0.99 (95% confidence interval [CI], 0.95 to 1.00) for the AmpliSens and Roche CAP/CTM Qual assays and 0.96 (95% CI, 0.90 to 0.98) for the Abbott Qualitative assay. The specificity was 1.00 (95% CI, 0.97 to 1.00) for the AmpliSens and Abbott Qualitative assays and 0.99 (95% CI, 0.96 to 1.00) for the Roche CAP/CTM Qual assay. McNemar analysis indicated that the proportions of positive results for the tests were not significantly different (P > 0.05). Cohen's kappa (0.97 to 0.99) indicated almost perfect agreement among the three tests. These results indicated that the AmpliSens DBS and whole-blood tests performed equally well and were comparable to the two commercially available EID tests. More importantly, the performance characteristics of the AmpliSens DBS test meets the World Health Organization EID test requirements; implementing AmpliSens DBS testing might improve EID services in resource-limited settings. Copyright © 2015, American Society for Microbiology. All Rights Reserved.

  6. Reverse transcription loop-mediated isothermal amplification for species-specific detection of tomato chlorotic spot orthotospovirus

    Science.gov (United States)

    Tomato chlorotic spot virus (TCSV) is an emerging tospovirus that can cause severe disease on tomato plants. There are at least four tospoviruses infecting tomato, and mixed infection of various viruses in a field crop is quite common. With similarity in the symptomatology and cross serological reac...

  7. Influencia do tabagismo isolado e associado a aspectos multifatoriais nos parametros acusticos vocais

    Directory of Open Access Journals (Sweden)

    Aline Gomes Lustosa Pinto

    2014-01-01

    Full Text Available Introdução: O fumo pode interferir negativamente na qualidade vocal e outros fatores associados, como o uso profissional da voz, hábitos, etilismo e refluxo gástrico-esofágico, podem potencializar essa interferência. O objetivo do estudo foi analisar a associação do tabagismo e dos demais fatores aos parâmetros acústicos vocais. Forma de estudo: Estudo de coorte contemporâneo com corte transversal. Materiais e métodos: Participaram do estudo 80 adultos, entre 35 e 60 anos, classificados nos grupos fumante (GF e controle (GC. Foi realizada aplicação de questionário e gravação das vozes. A avaliação acústica foi realizada com o software Praat. Foram utilizados os testes Mann-Whitney, qui-quadrado e regressão linear simples. Resultados: O GF apresentou maior prevalência de etilismo, tosse, hábito de pigarrear e uso profissional da voz, assim como apresentou valores aumentados para relação ruído-harmônico (NHR, jitter e shimmer. Foi estabelecida correlação entre o gênero feminino e todos os parâmetros acústicos, assim como entre o RGE, o aumento do jitter e o hábito de fumar, o agravamento da frequência fundamental, o aumento do jitter, o shimmer e a NHR. Conclusão: O fumo interfere nos parâmetros acústicos de modo isolado e/ou associado ao etilismo, tosse, hábito de pigarrear, RGE e uso profissional da voz.

  8. Variabilidade entre isolados de Trichoderma harzianum: I - Aspectos citológicos Variability among Trichoderma harzianum isolates: I - Cytological aspects

    Directory of Open Access Journals (Sweden)

    E. Peres

    1995-04-01

    Full Text Available Objetivou-se neste trabalho estudar a variabilidade de isolados selvagens de Trichoderma harzianum baseado nas características culturais e citológicas. Observaram-se o tamanho dos fialosporos, número de núcleos por fialosporos e crescimento e esporulação em meio de malte-ágar. Pelos resultados aqui encontrados foi possível reconhecer que há variação entre os isolados selvagens da espécie T. harzianum. Com relação ao número de núcleos, verificou-se uma variação de 1 a 3 núcleos por fialosporos. Também observou-se padrões diferenciais de crescimento e morfologia da colônia. Mais de 50% dos isolados atingiram o máximo de crescimento em 48 horas.This study is based largely on morphological and cultural characters of Tríchoderma harzianum isolates. It were observed the size of phialospores, mycelial growth and sporulation on malt extract agar and nuclei number per phialospores, stained with Giemsa. A x 100 oil immersion len was used in examining and in measuring phialospores. Based on the size of phialospores, it was possible recognize that there is variaton among the wild isolates for the specie T. harzianum. Also, with relation to mycelial growth and sporulation can itself distinguish from one another different pattern. Up to 50% of isolates had maximum growth in 48 hours. The nuclei number of 1 to 3 per phialospore was observed.

  9. Controle de Rhizoctonia solani e Fusarium oxysporum f.sp. phaseoli por biopreparados de isolados de Trichoderma spp.

    Directory of Open Access Journals (Sweden)

    Pedro Paulo Dias

    2013-12-01

    Full Text Available Os experimentos objetivaram avaliar em condições de casa de vegetação o biocontrole dos fitopatógenos Rhizoctonia solani (RS e Fusarium oxysporum f.sp. phaseoli (FOP em alface (Lactuca sativa L. cultivar Regina, e feijão-vagem (Phaseolus vulgaris L. cultivar Alessa, respectivamente, utilizando como agentes antagonistas, 10 isolados de Trichoderma spp. selecionados em testes in vitro. Foram feitos biopreparados à base de arroz previamente colonizado por isolados de Trichoderma spp. e posteriormente triturados. Para a realização dos testes, os biopreparados foram inoculados previamente na proporção de 10(9 conídios.mL-1, em substrato comercial para produção de mudas. Após sete dias, os patógenos foram introduzidos separadamente em duas concentrações distintas: R. solani na proporção de 144 mg de meio de arroz por kg de substrato e F. oxysporum f.sp. phaseoli inoculado na forma de suspensão contendo 4,75 x 10(6 conídios.mL-1. Avaliou-se a influência dos biopreparados na % de damping-off de pós-emergência em plantas de alface e a severidade de murcha em plantas de feijão-vagem. O biopreparado referente ao isolado T-03 foi o mais eficiente no controle de R. solani em plantas de alface cultivar Regina, por ter reduzido a incidência de damping-off de pós-emergência nessa cultura. Por outro lado, nenhum dos biopreparados apresentou efeito antagonista satisfatório à F. oxysporum f.sp. phaseoli em plantas de feijão-vagem.

  10. SENSIBILIDADE IN VITRO DE STAPHYLOCOCCUS AUREUS ISOLADOS DE AMOSTRAS DE LEITE DE VACAS COM MASTITE SUBCLÍNICA

    Directory of Open Access Journals (Sweden)

    Albenones José de Mesquita

    2006-10-01

    Full Text Available Foram submetidas a teste de sensibilidade in vitro a cinco antibióticos e quimioterápicos, 291 cepas de Staphylococcus aureus isolados de 667 amostras de leite procedentes de 375 vacas reagentes ao California Mastitis Test (CMT. Verificaram-se os seguintes percentuais de sensibilidade aos antimicrobianos: gentamicina, 90,72% (264/291; enrofloxacina, 89,35% (260/291; cefaperazona, 88,66% (258/291; kanamicina, 87,63% (255/291 e penicilina, 23,71% (69/291. PALAVRAS-CHAVE: Leite bovino, mastite subclínica, antimicrobianos.

  11. Produção e caracterização de celulases produzidas por um isolado de Bacillus sp.

    OpenAIRE

    Evilin Giordana de Marco

    2012-01-01

    Durante o processo de compostagem, a degradação de resíduos orgânicos é acompanhada e direcionada pela sucessão de populações microbianas que exibem uma ampla variedade de capacidades funcionais. Um total de 183 bactérias isoladas de uma leira de compostagem foram avaliadas quanto a capacidade de produção de celulases. Destas, após os primeiros ensaios, apenas um isolado foi selecionado para a realização dos testes de caracterização. Curvas de crescimento foram realizadas e em intervalos de 2...

  12. Isoflavonas em isolados e concentrados protéicos de soja Isoflavones in soy protein isolate and soy protein concentrate

    Directory of Open Access Journals (Sweden)

    Maria Cristina Y. Lui

    2003-12-01

    Full Text Available Isolados e concentrados protéicos de soja são ingredientes largamente utilizados na indústria de panificação, confeitaria, bebidas e embutidos. As isoflavonas presentes na soja podem sofrer alterações em quantidade e perfil de distribuição dependendo das condições de processamento. O objetivo deste estudo foi verificar o balanço de massa de isoflavonas e proteína em processamento de isolados e de concentrados protéicos de soja (tratamento com ácido e com álcool. A maior parte das isoflavonas presentes na matéria-prima (farinha desengordurada de soja é perdida nos sobrenadantes de processo (90% para extração com etanol 60%, 52% para processamento de isolado protéico e 47% para extração com ácido. O teor de isoflavonas nos produtos obtidos foi de 686µg/g base seca (b.s. para isolado protéico, 871µ g/g b.s. para concentrado protéico obtido por tratamento ácido e apenas 153µg/g b.s. para concentrado protéico obtido por tratamento com álcool. Não foi observada alteração no perfil de distribuição das isoflavonas nesse último processo, enquanto que nos dois primeiros notou-se diminuição da quantidade das formas malonil glicosídeos e aumento da quantidade das formas beta-glicosiladas e gliconas.Soy protein isolate (SPI and soy protein concentrate (SPC are largely used in bakery, confectionary, meat and beverage products. Isoflavones present in soybeans products can undergo changes in quantity and profile depending on the processing conditions. The objective of this work was to conduct mass balance studies of isoflavones and protein during the processing of SPI and SPC (acid and alcohol leach. The majority of isoflavones present in the raw material is lost in the supernatants (90% for SPC treated with alcohol, 52% for SPI and 47% for SPC treated with acid. Total concentration of isoflavones was 652µg/g for SPI, 838µg/g for SPC (acid leach, and only 147µg/g for SPC (alcohol leach. There were no changes in the

  13. Atividade antimicrobiana do extrato bruto de Açaí (Euterpe oleracea Mart.) em isolados clínicos

    OpenAIRE

    Alves, Maria José; Pimentel, Maria Helena; Baía, Lilia; Soares, Rafaela; Santos, Claudia; Wiliam C. B. Regis; Isabel C.F.R. Ferreira

    2016-01-01

    O presente trabalho teve como objetivo avaliar a atividade antimicrobiana do extrato de açaí (Euterpe Oleracea Mart.) em isolados clínicos com elevados perfis de resistência a diferentes antimicrobianos provenientes do Centro Hospitalar de Trás – os – Montes e Alto Douro – Unidade de Vila Real. A concentração mínima inibitória (CMI) foi determinada utilizando o ensaio colorimétrico rápido que utiliza o corante cloreto de p-iodonitrotetrazólio (INT) após já ter sido efetuado ...

  14. Analise do gas natural liquefeito como alternativa energetica para os pequenos e medios sistemas isolados da amazonia

    OpenAIRE

    Nelson Kuwahara

    1999-01-01

    Resumo: A política energética implementada na Amazônia tem favorecido fundamentalmente os grandes consumidores industriais, em detrimento do atendimento da população em geral. Verificam-se que as diretrizes do setor energético são determinadas pelas políticas de desenvolvimento econômico regional, e estas por sua vez estão quase sempre dissociadas das aptidões da região e descompromissadas com os aspectos sócio-ambientais. Os "sistemas isolados" de suprimento de energia elétrica na Amazônia s...

  15. Neurofibroma orbitário isolado e endotropia: relato de caso e revisão da literatura

    OpenAIRE

    Abbud, Christine Mae Morello; Cruz, Antonio Augusto Velasco e; Bicas,Harley Edison Amaral; Chaud, Fernando

    2003-01-01

    Os autores relatam um caso de neurofibroma orbitário, isolado, associado à endotropia do olho esquerdo, incomitante, de aparecimento lento e progressivo há 5 anos, sem referência à diplopia. O exame da sensorialidade mostrava acuidade visual normal em ambos os olhos e ausência de diplopia devido à supressão alternante. Exames de imagem da órbita (tomografia computadorizada e ressonância magnética) revelaram lesão alongada intracônica junto ao músculo reto lateral esquerdo. A paciente foi subm...

  16. CARACTERIZAÇÃO MOLECULAR DE ISOLADOS DE Colletotrichum spp. ASSOCIADOS A PODRIDÃO FLORAL DOS CITROS

    Directory of Open Access Journals (Sweden)

    AMANDA LETÍCIA DA SILVEIRA

    2016-02-01

    Full Text Available RESUMO A podridão floral dos citros (PFC é uma importante doença dessa cultura, responsável por elevadas perdas de produção. Normalmente, essa doença mostra-se limitante quando ocorrem prolongados períodos chuvosos durante o florescimento das plantas ou quando existe intenso molhamento foliar. Duas espécies de Colletotrichum estão associadas à doença: C. acutatum e C. gloeosporioides. Entretanto, recentemente, tem-se verificado que, mesmo sob condições não tão propícias, a doença tem ocorrido com relativa frequência, suspeitando-se do envolvimento de outras espécies de Colletotrichumou de novas condições de adaptação das espécies descritas. Este trabalho teve como objetivo determinar se há ou não outra espécie de Colletotrichum associada a PFC e avaliar a viabilidade do emprego de marcadores moleculares ISSR na caracterização taxonômica de isolados de Colletotrichum spp. associados a sintomas de PFC em flores, assim como de tecidos foliares e frutos cítricos assintomáticos. Para tanto, foi empregada uma combinação de iniciadores específicos, levando em conta a região ITS e marcadores moleculares ISSR. Os marcadores ISSR mostraram-se eficientes na caracterização taxonômica dos isolados de Colletotrichum analisados. A população avaliada foi constituída apenas por C. acutatum e C. gloeosporioides, descartando o envolvimento de uma espécie adicional. Foi constatada alta diversidade genética entre os isolados analisados, o que também se mostra convergente quanto às diferenças fenotípicas observadas sob condições de campo. Entretanto, não foi encontrada relação quanto à origem e as espécies de Colletotrichum spp. associadas. De modo inédito, ainda que assintomaticamente, foi detectada a presença de um isolado de C. acutatum associado a frutos cítricos.

  17. Natural incidence of tomato viruses in the North of Iran

    Directory of Open Access Journals (Sweden)

    Alireza Mohammadi HAJIABADI

    2012-09-01

    Full Text Available A survey was conducted in Qazvin province in the North of Iran, to determine the incidence of tomato viruses including: Tobacco mosaic virus (TMV, Tomato yellow leaf curl virus (TYLCV, Tomato chlorotic spot virus (TCSV, Tomato bushy stunt virus (TBSV, Tomato spotted wilt virus (TSWV, Tomato ring spot virus (ToRSV, Tomato aspermy virus (TAV, Potato virus Y (PVY, Beet curly top virus (BCTV, and Cucumber mosaic virus (CMV. A total of 742 tomato symptomatic samples were collected during the summer of 2007 in five regions of Qazvin province (Qazvin, Takestan, Boeen-Zahra, Alborz and Abiyek and tested by enzyme-linked immunosorbent assay (ELISA. TSWV was detected in Alborz (4.4 % and Abiyek (3.57% regions but TMV and CMV were detected in all five regions. The greatest and least incidence of tomato viruses were recorded in Alborz (40.7 % and Takestan (11.1 %, respectively. The presence of these viruses was also evaluated in the weed hosts as natural sources of plant viruses. The greatest and least incidence of tomato viruses in weed hosts were recorded in Boeen-Zahra (25.6 % and Qazvin (12.8 %, respectively. TSWV was not detected in weeds. Transmission tests demonstrated that Thrips tabaci acts as TSWV carrier and Myzus persicae and Aphis gossypii were CMV carriers. Seed transmission tests were positive for TMV (13 tomato seedlings from 100 seedlings, but no TSWV transmission was observed through the seeds of infected tomato fruits.

  18. Occurrence of Apple stem grooving virus in commercial apple seedlings and analysis of its coat protein sequence

    Science.gov (United States)

    Virus infections are responsible for reduced yield and quality in many crops, and are especially problematic in vegetatively-propagated crops such as apple. Three major viruses (Apple stem grooving virus (ASGV), Apple chlorotic leaf spot virus and Apple stem pitting virus) affect apple trees in Kore...

  19. Is this Red Spot the Blue Spot (locus ceruleum)?

    Energy Technology Data Exchange (ETDEWEB)

    Choe, Won Sick; Lee, Yu Kyung; Lee, Min Kyung; Hwang, Kyung Hoon [Gachon University Gil Hospital, Incheon (Korea, Republic of)

    2010-06-15

    The authors report brain images of 18F-FDG-PET in a case of schizophrenia. The images showed strikingly increased bilateral uptake in the locus ceruleum. The locus ceruleum is called the blue spot and known to be a center of the norepinephrinergic system.

  20. Survey of the Symptoms and Viruses Associated with Cowpea ...

    African Journals Online (AJOL)

    15.6%), necrotic spots (8.2%) and other symptoms (14.5%). The result of the laboratory assay to detect the viruses present in the leaf samples revealed the presence of four (4) different viruses. The viruses identified were, Cowpea Aphid Borne ...

  1. Imunofluorescência utilizando isolados brasileiros no diagnóstico sorológico de infecção por lentivírus em caprinos

    Directory of Open Access Journals (Sweden)

    Reischak Dilmara

    2002-01-01

    Full Text Available Os lentivírus de pequenos ruminantes (SRLV têm distribuição mundial e causam infecções persistentes em ovinos e caprinos. O objetivo deste trabalho foi desenvolver um teste de imunofluorescência indireta (IFA, utilizando isolados brasileiros de SRLV, para o diagnóstico sorológico de infecção por estes agentes em caprinos. A técnica de IFA foi comparada, quanto à sensibilidade e à especificidade, ao teste de AGID com antígeno do vírus Maedi-Visna WLC-1. Cultivos celulares secundários de membrana sinovial ovina infectadas com dois isolados de SRLV de origem caprina (CAEV Br/UFRGS-2 e CAEV Br/UFRGS-5 foram utilizados para o teste de IFA. Duzentas e trinta e nove amostras de soro caprino foram submetidas aos dois testes. O teste de AGID detectou 129 (53.9% amostras de soro caprino com anticorpos para SRLV. O teste de IFA detectou mais amostras reagentes, sendo que resultados diferentes foram observados de acordo com o isolado de SRLV empregado. Quando o isolado CAEV Br/UFRGS-2 foi utilizado como antígeno, 216 (90.3% amostras de soro caprino foram reagentes, enquanto que o isolado CAEV Br/UFRGS-5 detectou 213 (89.1% amostras de soro positivas. Não houve diferença estatisticamente significativa entre esses dois isolados. O teste de IFA desenvolvido teve sensibilidade de 94.6% e 96.9% e especificidade 14.5% e 20%, quando os isolados CAEV Br/UFRGS-2 e CAEV Br/UFRGS-5 foram usados como antígeno, respectivamente. O aprimoramento da técnica, assim como sua comparação com um teste mais sensível, ainda se fazem necessários. No entanto, os resultados demonstraram que a técnica de IFA, utilizando isolados brasileiros de SRLV como antígeno, apresenta potencial como um teste alternativo e complementar para o diagnóstico sorológico de infecção por estes agentes.

  2. Análise do funcionamento do gerador de ímanes permanentes, com anisotropia inversa, isolado, alimentando uma carga resistiva

    OpenAIRE

    Ferreira, Ângela P.

    2001-01-01

    A utilização da máquina de ímanes permanentes como gerador isolado tem vindo a ser estimulada pelo aumento das potências disponíveis, através do desenvolvimento dos materiais magnéticos permanentes, nomeadamente nos últimos vinte anos, caracterizado pelo aparecimento das ligas de terras raras e por uma selecção adequada dos parâmetros da máquina. A análise do funcionamento do gerador de ímanes permanentes isolado, alimentando uma carga resistiva, baseada no modelo de Blondel simplificado, per...

  3. Tomato chocolàte virus: a new plant virus infecting tomato and a proposed member of the genus Torradovirus

    NARCIS (Netherlands)

    Verbeek, M.; Dullemans, A.M.; Heuvel, van den J.F.J.M.; Maris, P.C.; Vlugt, van der R.A.A.

    2010-01-01

    A new virus was isolated from a tomato plant from Guatemala showing necrotic spots on the bases of the leaves and chocolate-brown patches on the fruits. Structural and molecular analysis showed the virus to be clearly related to but distinct from the recently described Tomato torrado virus (ToTV)

  4. RAS gene hot-spot mutations in canine neoplasias.

    Science.gov (United States)

    Richter, A; Murua Escobar, H; Günther, K; Soller, J T; Winkler, S; Nolte, I; Bullerdiek, J

    2005-01-01

    Point mutations in the cellular homologues HRAS, KRAS2, and NRAS of the viral Harvey and Kirsten rat sarcoma virus oncogenes are commonly involved in the onset of malignancies in humans and other species such as dog, mouse, and rat. Most often, three particular hot-spot codons are affected, with one amino acid exchange being sufficient for the induction of tumor growth. While RAS genes have been shown to play an important role in canine tumors such as non-small lung cell carcinomas, data about RAS mutations in canine fibrosarcomas as well as KRAS2 mutations in canine melanomas is sparse. To increase the number of tumors examined, we recently screened 13 canine fibrosarcomas and 11 canine melanomas for point mutations, particularly within the mutational hot spots. The results were compared to the already existing data from other studies about these tumors in dogs.

  5. Hot-spots in tapwaterleidingen

    NARCIS (Netherlands)

    Wolferen, J. van; Sluis, S.M. van der

    2002-01-01

    ln opdracht van de VNI is een aantal berekeningen uitgevoerd voor het vaststellen van aanvullende richtlijnen in verband met hot-spots in tapwaterleidingen. Hierbij is deels voortgebouwd op berekeningen die reeds eerder in opdracht van Novem zijn uitgevoerd t.b.v. ISSO publicatie 55.1, Handleiding

  6. Cassava Ivorian bacilliform virus is a member of the genus Anulavirus.

    Science.gov (United States)

    Scott, Simon W; MacFarlane, Stuart A; McGavin, Wendy J; Fargette, Denis

    2014-10-01

    The complete genomic sequence of Cassava Ivorian bacilliform virus (CIBV) is described. The virus has a genomic organization similar to that of pelargonium zonate spot virus (PZSV), the type member of the genus Anulavirus, but it is most closely related to a second, recently described, anulavirus, Amazon lily mild mottle virus (ALiMMV).

  7. Cassava Ivorian bacilliform virus is a member of the genus Anulavirus

    OpenAIRE

    Scott, S. W.; MacFarlane, S.A.; McGavin, W. J.; Fargette, Denis

    2014-01-01

    The complete genomic sequence of Cassava Ivorian bacilliform virus (CIBV) is described. The virus has a genomic organization similar to that of pelargonium zonate spot virus (PZSV), the type member of the genus Anulavirus, but it is most closely related to a second, recently described, anulavirus, Amazon lily mild mottle virus (ALiMMV).

  8. SPOT 4 North American Data Buy

    Data.gov (United States)

    U.S. Geological Survey, Department of the Interior — The USGS has contracted with SPOT Image Corporation to acquire and provide Satellite Pour l'Observation de la Terre (SPOT) satellite data for calendar years 2010 and...

  9. SPOT- 4 North American Data Buy

    Data.gov (United States)

    U.S. Geological Survey, Department of the Interior — The USGS has contracted with SPOT Image Corporation to acquire and provide Satellite Pour l'Observation de la Terre (SPOT) satellite data for calendar years 2010 and...

  10. SPOT- 5 North American Data Buy

    Data.gov (United States)

    U.S. Geological Survey, Department of the Interior — The USGS has contracted with SPOT Image Corporation to acquire and provide Satellite Pour l'Observation de la Terre (SPOT) satellite data for calendar years 2010 and...

  11. SPOT 5 North American Data Buy

    Data.gov (United States)

    U.S. Geological Survey, Department of the Interior — The USGS has contracted with SPOT Image Corporation to acquire and provide Satellite Pour l'Observation de la Terre (SPOT) satellite data for calendar years 2010 and...

  12. Laser Pyrometer For Spot Temperature Measurements

    Science.gov (United States)

    Elleman, D. D.; Allen, J. L.; Lee, M. C.

    1988-01-01

    Laser pyrometer makes temperature map by scanning measuring spot across target. Scanning laser pyrometer passively measures radiation emitted by scanned spot on target and calibrated by similar passive measurement on blackbody of known temperature. Laser beam turned on for active measurements of reflectances of target spot and reflectance standard. From measurements, temperature of target spot inferred. Pyrometer useful for non-contact measurement of temperature distributions in processing of materials.

  13. Highly Specific Detection of Five Exotic Quarantine Plant Viruses using RT-PCR

    OpenAIRE

    Hoseong Choi; Won Kyong Cho; Jisuk Yu; Jong-Seung Lee; Kook-Hyung Kim

    2013-01-01

    To detect five plant viruses (Beet black scorch virus, Beet necrotic yellow vein virus, Eggplant mottled dwarf virus, Pelargonium zonate spot virus, and Rice yellow mottle virus) for quarantine purposes, we designed 15 RT-PCR primer sets. Primer design was based on the nucleotide sequence of the coat protein gene, which is highly conserved within species. All but one primer set successfully amplified the targets, and gradient PCRs indicated that the optimal temperature for the 14 useful prime...

  14. Cylindrocladium spathiphylli de espatifilo (Spathiphyllum wallisii Rengel: detecção de enzimas extracelulares em isolados normais e alterados por temperatura

    Directory of Open Access Journals (Sweden)

    Bruna Cristina Migotto

    2013-06-01

    Full Text Available O fungo Cylindrocladium spathiphylli causa podridão no colo e nas raízes de espatifilo, induzindo amarelecimento e murcha nas folhas da parte aérea. Há poucos estudos com o fungo quanto às enzimas extracelulares. Assim, delineou-se ensaio inteiramente casualizado in vitro, com três tratamentos (dois isolados normais: LFEEI016 - EPAGRI/SC e MMBF 01/01 - IB/SP; mais testemunha e seis repetições, visando a detecção de enzimas. A amilase (AM, lipase (LP, carboximetilcelulase (CMC e lacase (LC foram mensuradas pelo cálculo da área da coroa circular, local de atividade das enzimas. A catalase (CT e gelatinase (GL foram mensuradas por símbolos, depois transformados em notas (1 a 4, de ausência à intensa produção. O isolado MMBF 01/01 produziu as maiores áreas e com mais intensidade as enzimas. Das enzimas, a maior área foi a da LC e depois a LP. LFEEI016 e MMBF 01/01 não apresentaram área para AM e CMC e exibiram intensidade fraca e igual para CT. A intensidade de GT foi moderada no MMBF 01/01 (nota média 3,0 e ausente para o LFEEI016 (nota média 1,0. Após a produção das enzimas extracelulares, os isolados foram alterados vegetativamente por temperatura. Instalou-se ensaio inteiramente ao acaso, em fatorial contendo disco de micélio dos isolados do fungo versus três temperaturas [23ºC (T1; 33ºC (T2 e de 33ºC para 23ºC (T3] versus quatro períodos (3; 6; 9 e 12 dias, com oito repetições, em placa de Petri com BDA mantida em BOD. O diâmetro das colônias foi medido diariamente (mm em dois sentidos. As alterações de temperatura vistas foram: inócuo, fungistático (FS e fungicida (FC. Aplicou-se estatística nas temperaturas T1 e T3 dos isolados para checar efeito FS e escolher aqueles com maior e menor efeito. O isolado LFEEIO16 mostrou efeito FS com três dias às temperaturas 33ºC e de 33ºC para 23ºC. Nos demais períodos, o isolado sofreu efeito FC. O MMBF 01/01 mostrou efeito FS em todos os períodos. Ambos

  15. Managing Rocky Mountain spotted fever.

    Science.gov (United States)

    Minniear, Timothy D; Buckingham, Steven C

    2009-11-01

    Rocky Mountain spotted fever is caused by the tick-borne bacterium Rickettsia rickettsii. Symptoms range from moderate illness to severe illness, including cardiovascular compromise, coma and death. The disease is prevalent in most of the USA, especially during warmer months. The trademark presentation is fever and rash with a history of tick bite, although tick exposure is unappreciated in over a third of cases. Other signature symptoms include headache and abdominal pain. The antibiotic therapy of choice for R. rickettsii infection is doxycycline. Preventive measures for Rocky Mountain spotted fever and other tick-borne diseases include: wearing long-sleeved, light colored clothing; checking for tick attachment and removing attached ticks promptly; applying topical insect repellent; and treating clothing with permethrin.

  16. Molecular characterization of Paecilomyces fumosoroseus (Deuteromycotina: Hyphomycetes isolates Caracterização molecular de isolados de Paecilomyces fumosoroseus (Deuteromycotina: Hyphomycetes

    Directory of Open Access Journals (Sweden)

    Andréia Cristiane Souza Azevedo

    2000-12-01

    Full Text Available ITS and RAPD analyses were used to investigate molecular variations within samples of Paecilomyces isolates and to resolve five morphologically atypical isolates resembling P. fumosorosus, obtained from whitefly in Northern Paraná State. The ITS4-ITS5 amplicon was 700 base pairs (bp long in all isolates. The five isolates of Paecilomyces not assigned to species produced restriction profiles identical to all the reference strains of P. fumosoroseus. The extent of fingerprint variability observed by RAPD was sufficient to discriminate all the isolates. The genetic similarity among unidentified isolates and strains of P. fumosoroseus was even higher than that observed among reference strains of this species, allowing us to conclude that isolates CNPso-P77, CNPso-P78, CNPso-P80, CNPso-P85 and CNPso-P91 are P. fumosoroseus.As análises de RAPD (Polimorfismo de DNA Amplificado ao Acaso e ITS (Sequências Internas Transcritas foram utilizadas para investigar a variabilidade molecular entre isolados de Paecilomyces e para identificar cinco isolados morfologicamente atípicos, obtidos a partir de mosca branca no Norte do Estado do Paraná, que possuíam alguma semelhança com P. fumosoroseus. O produto da amplificação com os primers ITS4 e ITS5 apresentou 700 pares de bases para todos os isolados investigados. Os cinco isolados não identificados deram origem a padrões de restrição idênticos a todas as linhagens de P. fumosoroseus utilizadas como referência. A variabilidade observada nos perfis de RAPD foi suficiente para discriminar todos os isolados. A similaridade genética entre os isolados não identificados e as linhagens de P. fumosoroseus foi maior do que aquela observada entre as linhagens referência desta espécie. Este fato permitiu concluir que os isolados CNPso-P77, CNPso-P78, CNPso-P80, CNPso-P85 and CNPso-P91 pertencem à espécie P. fumosoroseus.

  17. Ebola Virus and Marburg Virus

    Science.gov (United States)

    Ebola virus and Marburg virus Overview Ebola virus and Marburg virus are related viruses that cause hemorrhagic fevers — illnesses marked by severe bleeding (hemorrhage), organ failure and, in many ...

  18. Sweet Spots and Door Stops

    Science.gov (United States)

    Thompson, Michael; Tsui, Stella; Leung, Chi Fan

    2011-01-01

    A sweet spot is referred to in sport as the perfect place to strike a ball with a racquet or bat. It is the point of contact between bat and ball where maximum results can be produced with minimal effort from the hand of the player. Similar physics can be applied to the less inspiring examples of door stops; the perfect position of a door stop is…

  19. Caracterização de isolados de Rhizoctonia associados à queima foliar em Roraima = Characterization of Rhizoctonia isolates associated with foliar blight in Roraima.

    Directory of Open Access Journals (Sweden)

    Dayane Rodrigues Youssef

    2012-08-01

    Full Text Available O objetivo desse trabalho foi caracterizar isolados do fungo Rhizoctonia associados à queima foliar, obtidos de hospedeiros de importância econômica no estado de Roraima. Os isolados foram obtidos de plantas de feijão-caupi (Vigna unguiculata, soja (Glycine max, seringueira (Hevea brasiliensis, melancia (Citrullus lanatus, alface (Lactuca sativa e feijão-guandu (Cajanus cajan. Os parâmetros utilizados foram números de núcleos, grupo de anastomose e as características culturais da colônia, taxa de crescimento micelial e a formação de escleródios nos meios de cultura: batata dextrose agar (BDA, BDA+asparagina, BDA+extrato de levedura, Czapek Agar, maltose-peptona-agar, soil extract agar, sacarose-yeast-asparagina e V-8. Todos os 10 isolados estudados foram caracterizados como multinucleados e pertencentes à espécie Rhizoctonia solani. Três isolados de feijão-caupi, um de soja e o isolado de melancia foram identificados como AGI-1A e um isolado de feijãocaupi, um de soja e o isolado de feijão-guandu como AGI-1B. O isolado de seringueira não foi identificado como nenhum dos padrões de anastomose utilizado. Para a maioria dos isolados as maiores taxas de crescimento micelialforam obtidas no meio de cultura Soil Extract Agar. Dois tipos de escleródios, característicos do grupo AGI, foram observados: formação de 2-20 tufos placa-1 coloração variável, 1-2 mm e formação de 38-611 microescleródios placa-1, de coloração marrom, medindo 100 μm. A produção e o tipo de escleródio variaram com o isolado e o meio de cultura utilizado.The aim of this work was to characterize Rhizoctonia isolates associated with foliar blight symptom from hosts with economic importance at Roraima state. The isolates were recovered from cowpea (Vigna unguiculata, soybean (Glycine max, rubber tree (Hevea brasiliensis, watermelon (Citrullus lanatus, lettuce (Lactuca sativa and pigeonpea (Cajanus cajan. The evaluated characteristics were nuclear

  20. Justifications shape ethical blind spots.

    Science.gov (United States)

    Pittarello, Andrea; Leib, Margarita; Gordon-Hecker, Tom; Shalvi, Shaul

    2015-06-01

    To some extent, unethical behavior results from people's limited attention to ethical considerations, which results in an ethical blind spot. Here, we focus on the role of ambiguity in shaping people's ethical blind spots, which in turn lead to their ethical failures. We suggest that in ambiguous settings, individuals' attention shifts toward tempting information, which determines the magnitude of their lies. Employing a novel ambiguous-dice paradigm, we asked participants to report the outcome of the die roll appearing closest to the location of a previously presented fixation cross on a computer screen; this outcome would determine their pay. We varied the value of the die second closest to the fixation cross to be either higher (i.e., tempting) or lower (i.e., not tempting) than the die closest to the fixation cross. Results of two experiments revealed that in ambiguous settings, people's incorrect responses were self-serving. Tracking participants' eye movements demonstrated that people's ethical blind spots are shaped by increased attention toward tempting information. © The Author(s) 2015.

  1. Spot-to-Beam Procedure

    CERN Document Server

    Seidov, Zakir F; Yahalom, Asher

    2004-01-01

    We describe the interactive "STB" (spot_to_beam) MATHEMATICA procedure for a) approximating the spot image at the screen as ellipse, b) getting five parameters of the elliptic beam (two diameters, center coordinates, and orientation angle). The basic idea is to "map" the reference holes at screen onto the X-Y plane normal to the beam direction (Z-axis). All distortions of the image, e.g., due to camera-screen disposition can be, in principle, taken into account,assuming that the hole positions at screen and the orientation of the screen are known. With the non-linear LMS fitting, the "curved-coordinate-system" of the holes at image is transferred to the Cartesian coordinate system at XY-plane. Then the fitting ellipse is found in this latter system, by solving the system of N linear equations for 5 unknown parameters of beam ellipse, where N>5 is a number of reference points on edge of spot image. The examples of the real measurements at various screens will be demonstrated. The accuracy of beam diameters is ...

  2. Relações filogenéticas e diversidade de isolados de Guignardia spp oriundos de diferentes hospedeiros nas regiões ITS1-5,8S-ITS2 Phylogenetic relationships and diversity of Guignardia spp isolated from different hosts on ITS1-5,8S-ITS2 region

    Directory of Open Access Journals (Sweden)

    Ester Wickert

    2009-06-01

    Full Text Available Fungos do gênero Guignardia são frequentemente isolados em diferentes espécies de plantas, sendo muitas vezes caracterizados como fungos endofíticos. Entretanto, algumas espécies deste fungo, a exemplo de G. citricarpa e G. psidii, são causadores de importantes doenças que afetam culturas agrícolas, como a Mancha-Preta dos Citros (MPC e a podridão dos frutos de goiabeira, respectivamente. Também são apontados como causadores de manchas foliares em diferentes espécies de frutíferas e também em outras culturas. Este trabalho teve o objetivo de isolar, identificar e caracterizar a diversidade genética existente entre isolados de Guignardia oriundos de citros, mangueira, goiabeira, eucaliptos, jabuticabeira e pitangueira através da análise da sequência de DNA do cístron ITS1-5,8-S-ITS2. Verificou-se que os isolados obtidos pertencem às espécies G. citricarpa e G. mangiferae. Entretanto, dois grupos encontrados em mangueira não puderam ser identificados em nível de espécie com base em sua sequência de DNA em função da baixa similaridade com as sequências de diferentes espécies de Guignardia já depositadas em banco de dados. Desta forma, goiabeira, eucaliptos, jabuticabeira e pitangueira são hospedeiras de G. mangiferae, enquanto os citros hospedam duas formas, G. citricarpa e G. mangiferae. Já a mangueira é hospedeira de G. mangiferae e de dois outros grupos ainda não identificados. Verificou-se ainda que isolados de Guignardia obtidos de sintomas de podridão de fruto de goiabeira foram identificados como G. mangiferae.Fungi of Guignardia genera are commonly isolated from different plant species and most of the time they are characterized as endophytics. However, some species of this genus, as G. citricarpa and G. psidii are known as causal agents of serious diseases that affect cultures, such as the Citrus Black Spot and the guava fruit rot, respectively. They are also responsible for diseases that cause foliar

  3. Múltiplas estirpes de isolados de Mycobacteriumbovis identificados por tipagem molecular em bovinos abatidos em matadouros-frigoríficos

    Directory of Open Access Journals (Sweden)

    Fernando Alzamora Filho

    2014-02-01

    Full Text Available O objetivo do presente trabalho foi utilizar métodos bacteriológicos e moleculares para a identificação do Mycobacteriumbovis em lesões observadas em carcaças de bovinos durante a inspeção postmortem de rotina em matadouros-frigoríficos com serviço de inspeção oficial. Foi acompanhado o abate e a inspeção de 825.394 bovinos, sadios ao exame ante mortem pelo serviço de inspeção oficial em dez matadouros-frigoríficos do estado da Bahia. Carcaça de 180 bovinos apresentaram lesões sugestivas de tuberculose e por outras linfadenites. No isolamento bacteriano, 25 amostras apresentaram crescimento disgônico de colônias de coloração creme-amareladas em meio de cultura Stonebrink-Leslie. Desses isolados, 14 foram identificados como M. bovis PCR multiplex e pela técnica do spoligotyping foram discriminados oito diferentes espoligotipos do M. bovis, sendo sete descritos na literatura e um novo spoligotipo sem descrição anterior. O espoligotipo majoritário foi o SB0121, com cinco amostras, sendo descrito no Brasil e em outros países, seguidos por dois clusters, SB295 e SB1055, com dois isolados cada. O espoligotipo SB1145 e SB1648 foram referidos apenas no Brasil e Dinamarca, respectivamente. O espoligotipo SB140 já foi encontrado no Brasil, Argentina, Uruguai e Paraguai. Estes resultados demonstram que os espoligotipos obtidos são compartilhados, até o momento, entre estados brasileiros e entre países da América Latina e Europa. Sendo assim, a discriminação molecular de isolados de M. bovis através do Spoligotyping constitui-se numa ferramenta para estudos epidemiológicos da tuberculose bovina no Estado da Bahia.

  4. Diversidade e capacidade simbiótica de rizóbios isolados de nódulos de Mucuna-Cinza e Mucuna-Anã

    Directory of Open Access Journals (Sweden)

    Andréa Aparecida de Lima

    2012-04-01

    Full Text Available As espécies de mucuna são muito utilizadas como adubos verdes, e poucas informações estão disponíveis a respeito dos rizóbios nativos capazes de nodulá-las. O objetivo deste trabalho foi avaliar a diversidade e a capacidade simbiótica de isolados bacterianos de nódulos de mucuna-cinza (Mucuna pruriens (L. DC. e mucuna-anã (Mucuna deeringiana (Bort. Merr.. As bactérias foram isoladas de nódulos de mucunas cinza e anã cultivadas em vasos com solos de um sistema de produção agroecológica. Foram isoladas 160 bactérias, sendo 80 de mucuna-anã e 80 de mucuna-cinza, que foram autenticadas e selecionadas para avaliação da capacidade simbiótica. A diversidade dos isolados foi avaliada por meio das características culturais em meio de cultura YMA e da técnica de análise de restrição do produto de PCR do gene 16S rDNA. A inoculação de cinco isolados em mucuna-cinza e dois em mucuna-anã apresentou elevada biomassa da parte aérea. A maioria dos isolados apresentou crescimento rápido e acidificou o meio de cultura. A análise de restrição demonstrou que as bactérias isoladas apresentam baixa similaridade com estirpes de referência, sugerindo a existência de isolados pertencentes a novos grupos, capazes de nodular as mucunas anã e cinza.

  5. Patogenicidade, DL50 e TL50 de isolados de Metarhizium anisopliae (Metsch. Sorok. para o percevejo castanho das raízes Scaptocoris carvalhoi BECKER (Hemiptera: Cydnidae

    Directory of Open Access Journals (Sweden)

    Xavier Luciane Modenez Saldivar

    2005-01-01

    Full Text Available Objetivou-se neste trabalho avaliar a patogenicidade de isolados do fungo Metarhizium anisopliae em Scaptocoris carvalhoi BECKER, 1967, bem como determinar a Dose Letal média (DL50 e o Tempo Letal médio (TL50, em laboratório. Os experimentos foram conduzidos na Embrapa Agropecuária Oeste em Dourados, MS, durante 2003. Suspensões de quatro isolados de M. anisopliae (Ma7, Ma69, Ma283 e Ma342 foram preparadas nas concentrações 10(4, 10(5, 10(6, 10(7 e 10(8 conídios mL-1 e inoculados topicamente 5mil dessas suspensoes sobre S. carvalhoi correspondendo, respectivamente, a 50, 500, 5.000, 50.000 e 500.000 conídios percevejo-1. Após a inoculação, os insetos (10 adultos e cinco ninfas foram acondicionados em gerbox (parcela e mantidos em câmaras climatizadas reguladas para 26?1masculineC, UR 85%, sem fotofase. Na maior dose testada (500.000 conídios inseto-1, observou-se um incremento significativo de mortalidade do percevejo com o aumento da dose do fungo, para todos os isolados testados. Os menores valores da DL50 foram observados com os isolados Ma69 e Ma7 e o maior com Ma283. Os valores do TL50 variaram de 0,32 a 5,84 dias, sem diferirem significativamente, entre si. Os isolados Ma69 e Ma7 apresentam potencial para serem empregados no controle de S. carvalhoi a campo.

  6. HIDROFOBICIDADE DE RIBOTIPOS DE BACILLUS CEREUS ISOLADOS DE INDÚSTRIA DE LATICÍNIOS

    Directory of Open Access Journals (Sweden)

    Emiliane Andrade ARAÚJO

    2010-03-01

    Full Text Available

    A contaminação de superfícies por microrganismos deterioradores e patogênicos é causa de preocupação na indústria de alimentos. Desenvolvimento de biofilmes em ambientes de processamento de alimentos resulta na deterioração do produto e em possíveis riscos para a saúde pública, além de criar sérios problemas nas operações de processamento do fl uido. A adesão da bactéria à superfície é um dos primeiros passos para a formação do biofilme e propriedades físico-químicas da interface bacteriana infl uenciam o processo de adesão microbiana e, consequentemente, os procedimentos operacionais de higienização. A estrutura do biofilme e as características fisiológicas do microrganismo podem conferir resistência aos agentes antimicrobianos, como por exemplo, aos sanitizantes usados no procedimento de higienização. Dentre os fatores que infl uenciam o processo de adesão, as características de hidrofobicidade do microrganismo e da superfície apresentam grande importância nos mecanismos de adesão. As técnicas da medição do ângulo de contato e da coluna de interação hidrofóbica, para determinação da hidrofobicidade de superfícies de ribotipos de Bacillus cereus isolados de indústria de laticínios, foram avaliadas. Observou-se que as quatro superfícies dos ribotipos avaliados apresentaram a mesma classificação quanto à hidrofobicidade tanto pela medida do ângulo de contato com a água quanto pela determinação da energia livre de interação hidrofóbica (ΔGsas TOT. Três ribotipos foram considerados hidrofílicos e um hidrofóbico. Já a técnica da coluna de interação hidrofóbica não mostrou diferença (p>0,05 no percentual de células retidas na coluna, sugerindo que as superfícies dos ribotipos apresentam as mesmas características quanto à hidrofobicidade. Os resultados indicam que a técnica da medição do ângulo de contato é a mais indicada para avaliar a

  7. First report of Tobacco rattle virus in spinach in California.

    Science.gov (United States)

    In 2009 in coastal California (Santa Barbara County), commercially grown spinach (Spinacia oleracea) in two nearby fields exhibited symptoms of a previously unrecognized virus-like disease. Symptoms consisted of general chlorosis and bright yellow blotches and spots. Necrotic spots were also associa...

  8. [Clinical value of cupping spot effect].

    Science.gov (United States)

    Ji, Chang-Chun; Huang, Li-Ping; Yang, Gai-Qin; Zhao, Jing-Yu; Zou, Wei; Guo, Xiao-Chuan; Liu, Shao-Ming

    2014-12-01

    The cupping spot is considered as one kind of skin change due to cupping treatment. With literature regarding cupping spot, the influencing factors and value of cupping spot in clinical diagnosis and treatment were analyzed, which could make a further exploration on the action mechanism of cupping treatment. The literature showed that the formation of cupping spot was related with cupping temperature, pressure, cup-retaining time, cupping area, individual difference and health condition, etc; cupping spot had the ability to assist diagnosis, prevent disease, cure disease and evaluate clinical efficacy. Previous studies on cupping spot have already made some progress, and played a positive significance on finding cupping rule and studying its mechanism. However, the research for this area is still in the primary stage, which needed deeper study to reveal scientific connotations of cupping spot.

  9. A multiple reverse transcription-polymerase chain reaction assay for simultaneous detection and differentiation of latent viruses and apscarviroids in apple trees

    Science.gov (United States)

    Apple chlorotic leaf spot virus (ACLSV), Apple stem grooving virus (ASGV), and Apple stem pitting virus (ASPV) are three latent viruses frequently occurring in apple trees worldwide. In field orchards, these viruses are frequently found in a mixed infection with viroids in the genus Apscarviroid, in...

  10. Efeito do fosfito de potássio isolado e em mistura com fungicidas no controle da requeima do tomateiro

    OpenAIRE

    Töfoli,J.G.; Mello,S.C.; Domingues,R.J.

    2012-01-01

    Três experimentos foram realizados, um em casa-de-vegetação e laboratório e os demais em campo, com o objetivo de estudar a ação do fosfito de potássio isolado e em mistura com fungicidas no controle de Phytophthora infestans do tomateiro. A aplicação de 300 g 100 L-1 de fosfito de potássio associado com mancozebe foi mais eficiente no controle da requeima nos cultivares 'Rebeca' e 'Giuliana', em relação à aplicação isolada desses produtos, no quarto e sétimo dia após a inoculação em discos f...

  11. Resistência antimicrobiana de coliformes isolados de leite humano ordenhado Antimicrobial resistance of coliform isolates from expressed human milk

    Directory of Open Access Journals (Sweden)

    Franz Reis Novak

    2001-06-01

    Full Text Available A veiculação de microrganismos potencialmente patogênicos, resistentes aos antibióticos, por meio de leite humano ordenhado, pode ser um fator de risco potencial. Este estudo teve como objetivo contribuir para um melhor conhecimento dos coliformes isolados de leite humano ordenhado (LHO e de seu perfil de resistência a antimicrobianos. Seguiu-se um delineamento inteiramente casualizado, utilizando 837 amostras de LHO, nas quais 71 (8,48% estavam contaminadas com coliformes totais, cuja contagem em nenhuma das amostras ultrapassava 1,0x10³NMP/ml. A maioria dos microrganismos isolados (91,6% pertencia a apenas duas espécies, Enterobacter cloacae e Klebsiella pneumoniae. As 71 cepas de coliformes apresentavam resistência a pelo menos um dos antimicrobianos testados. Conclui-se que o LHO coletado e armazenado sob condições higiênico-sanitárias insatisfatórias, pode apresentar coliformes, que desgastam os fatores de proteção, reduzem o valor nutricional e a qualidade do produto.The dispersion of potentially pathogenic, antibiotic-resistant microorganisms via expressed human milk can be considered a risk factor. The aim of this study was to contribute to a better understanding of coliform isolates from expressed human milk and their antimicrobial resistance profiles. The sampling scheme followed a totally randomized design, using 837 samples of expressed human milk. Of these, 71 (8.48% were identified as contaminated with total coliforms, although in none of the samples did the population exceed 1.0x10³ MPN/ml. Most of the microorganisms isolated (91.6% belonged to only two species, Enterobacter cloacae and Klebsiella pneumoniae, which when subjected to antibiograms, revealed that several strains showed prior resistance to some of the antimicrobials tested. Coliforms may grow in expressed human milk if it is improperly stored, depleting protection factors and reducing the milk's nutritional value.

  12. Reduction of incidence and severity of Septoria lycopersici leaf spot of tomato with bacteria and yeasts Redução da incidência e severidade da mancha foliar do tomateiro causada por Septoria lycopersici com bacteria e leveduras

    Directory of Open Access Journals (Sweden)

    Luiz Eduardo Bassay Blum

    2000-10-01

    Full Text Available Septoria leaf spot, caused by Septoria lycopersici, is an important disease of tomato (Lycopersicon esculentum which is mainly controlled by fungicide sprays. One of the alternatives to reduce fungicide applications is the use of leaf antagonists such as yeast and bacterium. This study was conducted from 1994 through 1995 in Auburn, AL, USA. The pathogen and one antagonist were isolated from field plants. In greenhouse, six yeast and one bacterial isolates were tested, in a set of seven experiments. The experiments were conducted in a completely randomized design with four to eight treatments and six replications. The antagonists (1-3 × 10(8 colony forming units ml-1 were inoculated 48h before the inoculation of the pathogen (1-2 × 10(5 conidia ml-1, under conditions of intermittent misting. The yeast isolate Y236 (Cryptococcus laurentii and the bacterial isolate BTL (Pseudomonas putida significantly (P A mancha foliar causada pelo fungo Septoria lycopersici é uma doença no tomateiro (Lycopersicon esculentum, controlada basicamente pela aplicação de fungicidas. Uma das alternativas ao controle químico dessa enfermidade é a utilização de bactérias e leveduras antagonistas. Este estudo foi conduzido de 1994 a 1995 em Auburn, Alabama, EUA. O patógeno e um dos antagonistas foram isolados do filoplano de plantas de tomateiro infectadas pela doença em questão. Sete antagonistas (um isolado de bacteria e seis de leveduras foram testados em uma série de sete experimentos conduzidos em casa de vegetação. Os experimentos foram conduzidos em um delineamento completamente casualizado com quatro a oito tratamentos e seis repetições. Em todos os experimentos, os antagonistas (1-3 × 10(8 unidades formadoras de colonia ml-1 foram inoculados 48h antes da inoculação com o patógeno (1-2 × 10(5 conidios ml-1, sob condições de nebulosidade intermitente. Entre os antagonistas testados, destacaram-se o isolado de levedura Y236 (Cryptococcus

  13. Variabilidade genética entre isolados de Colletotrichum gossypii do algodoeiro Genetic variability among the isolates of Colletotrichum gossypii of cotton

    Directory of Open Access Journals (Sweden)

    Yeshwant R. Mehta

    2010-03-01

    Full Text Available O algodoeiro é atacado por Colletotrichum gossypii (CG e C. gossypii var. cephalosporioides (CGC. Ambos os patógenos são transmitidos pela semente e sua distinção morfológica é extremamente difícil e inconsistente. Tentativas foram feitas no presente trabalho para verificar a variabilidade genética entre CG e CGC através de RAPD-PCR, ERIC- e REP-PCR e PCR-RFLP da região ITS rDNA. Foram utilizados 53 isolados coletados de sementes e folhas de plantas de diferentes cultivares nos estados do Paraná, São Paulo, Mato Grosso, Minas Gerais, e Paraiba, entre 1999 e 2003. Baseado em testes de patogenicidade, vinte e um isolados foram classificados como CG e 32 como CGC. Os resultados obtidos por RAPD-PCR, utilizando-se oito primers, revelaram dois grupos distintos sendo que o primeiro foi formado por 94% dos isolados de sementes e o segundo por 95% dos isolados de folhas. Na análise de ERIC- e REP-PCR, resultados semelhantes a RAPD foram obtidos, sendo que o primeiro grupo foi formado por 93% dos isolados provenientes das sementes e o segundo por 78% dos isolados provenientes das folhas. Quando o produto de amplificação da região ITS rDNA foi digerido com oito enzimas de restrição, um perfil de bandas semelhante para todos os isolados foi obtido. Resultados de RAPD, ERIC- e REP-PCR demonstraram que existem diferenças genéticas entre os isolados provenientes das sementes e aqueles provenientes de parte aérea, e esses dois grupos foram claramente distintos. Estudos futuros devem ser realizados utilizando outras técnicas moleculares para a obtenção de marcadores capazes de distinguir entre isolados de CG e CGC.Cotton is attacked by Colletotrichum gossypii (CG and C. gossypii var. cephalosporioides (CGC. Both the pathogens are transmitted by seed and their morphological distinction is extremely difficult and inconsistent. In the present study, attempts were made to verify the genetic variability among 53 isolates of CG and CGC using

  14. AGGRESSIVENESS AND EXOENZIMES PRODUCTION OF Colletotrichum ISOLATES ON ORNAMENTAL TROPICAL PLANTS AGRESSIVIDADE E PRODUÇÃO DE EXOENZIMAS DE Colletotrichum ISOLADOS DE PLANTAS ORNAMENTAIS TROPICAIS

    Directory of Open Access Journals (Sweden)

    Rildo Sartori Barbosa Coelho

    2011-04-01

    Full Text Available Anthracnose, generally caused by Colletotrichum gloeosporioides, affects inflorescences quality in different species of ornamental tropical plants and it is one of the main fungal diseases to those crops. The objective of this study was to evaluate the aggressiveness and exoenzimes production on solid medium of  25 Colletotrichum isolates collected from anthurium, torch ginger, and heliconia plants with necrosis symptoms. The symptomatic floral stems were harvested in different counties of the Pernambuco State, Brazil, and carried to the laboratory, where the isolation process took place. The isolates were inoculated in the floral stems bracts of the three species studied, and the aggressiveness evaluated five days after inoculation. Isolates were also plated on specific culture media to determine amylolytic, lipolytic, and proteolytic activity, and assessed seven days after inoculation. The isolates aggressiveness was variable and some of them were not pathogenic when inoculated to their origin hosts. Six isolates were pathogenic to all the analyzed ornamental species and four isolates showed host specificity. All tested isolates presented amylolytic, lipolytic, and proteolytic activity. No relation between aggressiveness and enzyme activity was observed in the isolates tested.

    KEY-WORDS: Anthurium andraeanum; Etlingera elatior; Heliconia spp.; variability.

    A antracnose, geralmente causada por Colletotrichum gloeosporioides, afeta a qualidade de inflorescências em diversas espécies de plantas ornamentais tropicais e destaca-se como uma das principais doenças de origem fúngica, para estas culturas. O objetivo deste trabalho foi verificar a agressividade e a produção de exoenzimas, em meio sólido, de 25 isolados de Colletotrichum obtidos a partir de inflorescências de

  15. Watermarking spot colors in packaging

    Science.gov (United States)

    Reed, Alastair; Filler, TomáÅ.¡; Falkenstern, Kristyn; Bai, Yang

    2015-03-01

    In January 2014, Digimarc announced Digimarc® Barcode for the packaging industry to improve the check-out efficiency and customer experience for retailers. Digimarc Barcode is a machine readable code that carries the same information as a traditional Universal Product Code (UPC) and is introduced by adding a robust digital watermark to the package design. It is imperceptible to the human eye but can be read by a modern barcode scanner at the Point of Sale (POS) station. Compared to a traditional linear barcode, Digimarc Barcode covers the whole package with minimal impact on the graphic design. This significantly improves the Items per Minute (IPM) metric, which retailers use to track the checkout efficiency since it closely relates to their profitability. Increasing IPM by a few percent could lead to potential savings of millions of dollars for retailers, giving them a strong incentive to add the Digimarc Barcode to their packages. Testing performed by Digimarc showed increases in IPM of at least 33% using the Digimarc Barcode, compared to using a traditional barcode. A method of watermarking print ready image data used in the commercial packaging industry is described. A significant proportion of packages are printed using spot colors, therefore spot colors needs to be supported by an embedder for Digimarc Barcode. Digimarc Barcode supports the PANTONE spot color system, which is commonly used in the packaging industry. The Digimarc Barcode embedder allows a user to insert the UPC code in an image while minimizing perceptibility to the Human Visual System (HVS). The Digimarc Barcode is inserted in the printing ink domain, using an Adobe Photoshop plug-in as the last step before printing. Since Photoshop is an industry standard widely used by pre-press shops in the packaging industry, a Digimarc Barcode can be easily inserted and proofed.

  16. Caracterização de variantes de Grapevine Fanleaf Virus (GFLV), Arabis Mosaic Virus (ARMV) e respectivos RNAS satélites presentes em castas portuguesas de Vitis Vinifera

    OpenAIRE

    Reis, Rita Alexandra Feliciano dos

    2015-01-01

    Dissertação de mestrado, Biologia Molecular e Microbiana, Faculdade de Ciências e Tecnologia, Universidade do Algarve, 2015 Grapevine fanleaf virus, vírus do urticado ou nó-curto da videira e Arabis mosaic virus são dois Nepovirus, da família Secoviridae e ambos possuem um genoma bipartido de cadeia simples e sentido positivo. Além destes, foram detetados em alguns isolados de ArMV e GFLV, RNAs satélite de grande tamanho, satRNA do tipo B. Estes vírus encontram-se entre os principais agent...

  17. White Spot Syndrome Virus infection in Penaeus monodon is ...

    Indian Academy of Sciences (India)

    2013-11-06

    Nov 6, 2013 ... 2007) and arginine kinase (Rattanarojpong et al. 2007; Somboonwiwat et al. 2010) in WSSV infection. It has been reported that cellular metabolic proteins have additional roles in immunity and transcriptional regulation of apoptosis (Kim and Dang. 2005). Externalization of glycolytic enzymes is a common.

  18. Molecular variability analyses of Apple chlorotic leaf spot virus ...

    Indian Academy of Sciences (India)

    Individual recombination analysis for all the pome and stone fruit isolates was also carried out. Plum isolates indicated possible minor parenting by Indian isolates. (India17, 18) for P863 (AAA42589), PBM1 (CAB46654) and major parenting for SX/2 (AAF67188), French (NP_. 040553) isolates with SX/2 and P863 isolates, ...

  19. Virus diseases in lettuce in the Mediterranean basin.

    Science.gov (United States)

    Moreno, Aranzazu; Fereres, Alberto

    2012-01-01

    Lettuce is frequently attacked by several viruses causing disease epidemics and considerable yield losses along the Mediterranean basin. Aphids are key pests and the major vectors of plant viruses in lettuce fields. Lettuce mosaic virus (LMV) is probably the most important because it is seed-transmitted in addition to be transmissible by many aphid species that alight on the crop. Tomato spotted wilt virus (TSWV) is another virus that causes severe damage since the introduction of its major vector, the thrips Frankliniella occidentalis. In regions with heavy and humid soils, Lettuce Mirafiori big-vein virus (LMBVV) can also produce major yield losses. Copyright © 2012 Elsevier Inc. All rights reserved.

  20. Integrating sustainable hunting in biodiversity protection in Central Africa: hot spots, weak spots, and strong spots.

    Directory of Open Access Journals (Sweden)

    John E Fa

    Full Text Available Wild animals are a primary source of protein (bushmeat for people living in or near tropical forests. Ideally, the effect of bushmeat harvests should be monitored closely by making regular estimates of offtake rate and size of stock available for exploitation. However, in practice, this is possible in very few situations because it requires both of these aspects to be readily measurable, and even in the best case, entails very considerable time and effort. As alternative, in this study, we use high-resolution, environmental favorability models for terrestrial mammals (N = 165 in Central Africa to map areas of high species richness (hot spots and hunting susceptibility. Favorability models distinguish localities with environmental conditions that favor the species' existence from those with detrimental characteristics for its presence. We develop an index for assessing Potential Hunting Sustainability (PHS of each species based on their ecological characteristics (population density, habitat breadth, rarity and vulnerability, weighted according to restrictive and permissive assumptions of how species' characteristics are combined. Species are classified into five main hunting sustainability classes using fuzzy logic. Using the accumulated favorability values of all species, and their PHS values, we finally identify weak spots, defined as high diversity regions of especial hunting vulnerability for wildlife, as well as strong spots, defined as high diversity areas of high hunting sustainability potential. Our study uses relatively simple models that employ easily obtainable data of a species' ecological characteristics to assess the impacts of hunting in tropical regions. It provides information for management by charting the geography of where species are more or less likely to be at risk of extinction from hunting.

  1. Vegetative compatibility and molecular characterization of Fusarium graminearum isolates from the State of Paraná, Brazil Compatibilidade vegetativa e caracterização molecular de isolados patogênicos de Fusarium graminearum do Estado do Paraná

    Directory of Open Access Journals (Sweden)

    Cleverson Busso

    2007-12-01

    Full Text Available Fusarium graminearum isolates causing Fusarium head blight in wheat were collected in Brazil and analyzed by random amplified polymorphic DNA (RAPD markers and vegetative compatibility grouping (VCG. Nitrate non-utilizing mutants (nit from each isolate were paired to verify heterokaryon formation. Three VCGs were identified among F. graminearum isolates: VCG1 included F-2, F-3 and F-4 isolates; VCG2 included F-1, F-6 and F-9 isolates; VCG3 included F-5, F-7 and F-8 isolates. Based on PCR amplification with eight different primers, the isolates showed great genetic similarity among themselves. Dendrogram analysis demonstrated two RAPD groups: Group A, consisting of isolates F-2 and F-9, and Group B, composed of the remaining isolates. Results suggest the clonal origin of F. graminearum isolates.Isolados de Fusarium graminearum, obtidos de espigas de trigo com sintomas de Giberela, foram analisados pela técnica do Polimorfismo de DNA Amplificado ao Acaso (RAPD e pelos Grupos de Compatibilidade Vegetativa (GCV. Mutantes auxotróficos (nit de cada isolado foram pareados em todas as combinações possíveis, para a formação de heterocários. Três GCVs foram identificados: GCV1, incluindo os isolados F-2, F-3 e F-4; GCV2, incluindo os isolados F-1, F-6 e F-9; e GCV3, formado pelos isolados F-5, F-7 e F-8. Dois grupos foram identificados com base nos marcadores de RAPD: o grupo A, formado pelos isolados F-2 e F-9, e o grupo B, composto pelos demais isolados, os quais apresentaram grande similaridade entre si. Os resultados sugerem a origem clonal dos isolados de F. graminearum analisados.

  2. HotSpot Wizard: a web server for identification of hot spots in protein engineering

    National Research Council Canada - National Science Library

    Pavelka, Antonin; Chovancova, Eva; Damborsky, Jiri

    2009-01-01

    HotSpot Wizard is a web server for automatic identification of 'hot spots' for engineering of substrate specificity, activity or enantioselectivity of enzymes and for annotation of protein structures...

  3. Identificação molecular de isolados de fungos de interesse médico por meio de marcadores RAPD - doi: 10.5102/ucs.v8i2.1110

    Directory of Open Access Journals (Sweden)

    Anabele Azevedo Lima

    2011-04-01

    Full Text Available Aspergillus e Penicillium são microrganismos capazes de provocar complicações clínicas consideráveis principalmente em pacientes imunossuprimidos. A análise molecular é uma estratégia importante para a identificação rápida e precisa desses agentes patogênicos. O objetivo desse trabalho foi descrever padrões de marcadores RAPD específicos para a identificação de isolados de Aspergillus e Penicillium causadores de doenças de interesse médico. A análise morfológica revelou que 14 isolados eram do gênero Aspergillus, 7 eram Penicillium, 1 correspondente a Fusarium e um isolado não foi identificado. A análise por RAPD com dez iniciadores produziram fragmentos de DNA característicos para cada gênero. O dendrograma revelou grupos de isolados dentro de cada gênero de fungo de acordo com perfis de marcadores específicos, identificando isolados muito semelhantes que apresentaram alto coeficiente de similaridade. Esses dados fornecem opções para o desenvolvimento de oligonucleotídios de PCR para a identificação específica de isolados específicos ocorrendo no ambiente médico-hospitalar.

  4. Genetic variability of HIV-1 isolates from Minas Gerais, Brazil Variabilidade genética de isolados de HIV-1 em Minas Gerais, Brasil

    Directory of Open Access Journals (Sweden)

    Anna Bárbara de Freitas Carneiro Proietti

    1999-04-01

    Full Text Available We report results of nucleotide sequencing and phylogenetic analysis of the env gene of 11 HIV-1 isolates, in Belo Horizonte, Brazil. Ten isolates belonged to HIV-1 subtype B and one was a probable B/F mosaic. This putative B/F recombinant is similar but not identical in its nucleotide sequence to other B/F mosaics described in Brazil.Relatamos resultados do estudo de seqüência de nucleotídeos e análise filogenética do gene env 11 isolados HIV-1 em Belo Horizonte, Brasil. Dez isolados pertenciam ao subtipo B e um era provavelmente um mosaico B/F. Este possível recombinante B/F é similar, mas não idêntico, em sua seqüência de nucleotídeos, aos demais mosaicos B/F descritos no Brasil.

  5. Padronização de modelo de coração isolado "working heart" com circulação parabiótica

    OpenAIRE

    Silveira Filho,Lindemberg da Mota; Petrucci Junior,Orlando; Carmo,Marcio Roberto do; Oliveira,Pedro Paulo Martins de; Vilarinho,Karlos Alexandre de Sousa; Vieira,Reinaldo Wilson; Braile,Domingo Marcolino

    2008-01-01

    OBJETIVO: Desenvolver modelo de coração isolado de suíno "working heart" sob suporte por circulação parabiótica e verificar se o mesmo é estável e se possibilitou de forma efetiva a mensuração dos dados propostos. MÉTODOS: O modelo foi padronizado durante preparação para estudo de associação de agente à solução cardioplégica. Foram realizados 18 experimentos com um animal suporte e um animal doador em cada. O coração do animal doador foi perfundido como coração isolado pelo animal suporte em ...

  6. Atividade celulolítica de fungos isolados de bagaço de cana-de-açúcar e madeira em decomposição.

    OpenAIRE

    Thalita Peixoto Basso; Cláudio Rosa Gallo; Luiz Carlos Basso

    2011-01-01

    O objetivo deste trabalho foi identificar isolados de fungos a partir de bagaço de cana-de-açúcar e madeira em decomposição e avaliar a sua atividade celulolítica em bagaço de cana. Cinco isolados foram avaliados, tendo-se como referências os fungos Trichoderma reesei QM9414 e T. reesei RUT C30. A atividade celulolítica foi estimada pela capacidade hidrolítica do extrato enzimático dos fungos cultivados em bagaço de cana sobre os substratos papel de filtro (atividade celulolítica total) e car...

  7. Unsupervised image segmentation for microarray spots with irregular contours and inner holes.

    Science.gov (United States)

    Belean, Bogdan; Borda, Monica; Ackermann, Jörg; Koch, Ina; Balacescu, Ovidiu

    2015-12-23

    Microarray analysis represents a powerful way to test scientific hypotheses on the functionality of cells. The measurements consider the whole genome, and the large number of generated data requires sophisticated analysis. To date, no gold-standard for the analysis of microarray images has been established. Due to the lack of a standard approach there is a strong need to identify new processing algorithms. We propose a novel approach based on hyperbolic partial differential equations (PDEs) for unsupervised spot segmentation. Prior to segmentation, morphological operations were applied for the identification of co-localized groups of spots. A grid alignment was performed to determine the borderlines between rows and columns of spots. PDEs were applied to detect the inflection points within each column and row; vertical and horizontal luminance profiles were evolved respectively. The inflection points of the profiles determined borderlines that confined a spot within adapted rectangular areas. A subsequent k-means clustering determined the pixels of each individual spot and its local background. We evaluated the approach for a data set of microarray images taken from the Stanford Microarray Database (SMD). The data set is based on two studies on global gene expression profiles of Arabidopsis Thaliana. We computed values for spot intensity, regression ratio, and coefficient of determination. For spots with irregular contours and inner holes, we found intensity values that were significantly different from those determined by the GenePix Pro microarray analysis software. We determined the set of differentially expressed genes from our intensities and identified more activated genes than were predicted by the GenePix software. Our method represents a worthwhile alternative and complement to standard approaches used in industry and academy. We highlight the importance of our spot segmentation approach, which identified supplementary important genes, to better explains

  8. Cosmicflows-3: Cold Spot Repeller?

    Science.gov (United States)

    Courtois, Hélène M.; Tully, R. Brent; Hoffman, Yehuda; Pomarède, Daniel; Graziani, Romain; Dupuy, Alexandra

    2017-09-01

    The three-dimensional gravitational velocity field within z ˜ 0.1 has been modeled with the Wiener filter methodology applied to the Cosmicflows-3 compilation of galaxy distances. The dominant features are a basin of attraction and two basins of repulsion. The major basin of attraction is an extension of the Shapley concentration of galaxies. One basin of repulsion, the Dipole Repeller, is located near the anti-apex of the cosmic microwave background dipole. The other basin of repulsion is in the proximate direction toward the “Cold Spot” irregularity in the cosmic microwave background. It has been speculated that a vast void might contribute to the amplitude of the Cold Spot from the integrated Sachs-Wolfe effect.

  9. Cosmicflows-3: Cold Spot Repeller?

    Energy Technology Data Exchange (ETDEWEB)

    Courtois, Hélène M.; Graziani, Romain; Dupuy, Alexandra [University of Lyon, UCB Lyon 1, CNRS/IN2P3, IPN, Lyon (France); Tully, R. Brent [Institute for Astronomy, University of Hawaii, 2680 Woodlawn Drive, Honolulu, HI 96822 (United States); Hoffman, Yehuda [Racah Institute of Physics, Hebrew University, Jerusalem, 91904 (Israel); Pomarède, Daniel [Institut de Recherche sur les Lois Fondamentales de l’Univers, CEA, Université Paris-Saclay, F-91191 Gif-sur-Yvette (France)

    2017-09-20

    The three-dimensional gravitational velocity field within z ∼ 0.1 has been modeled with the Wiener filter methodology applied to the Cosmicflows-3 compilation of galaxy distances. The dominant features are a basin of attraction and two basins of repulsion. The major basin of attraction is an extension of the Shapley concentration of galaxies. One basin of repulsion, the Dipole Repeller, is located near the anti-apex of the cosmic microwave background dipole. The other basin of repulsion is in the proximate direction toward the “Cold Spot” irregularity in the cosmic microwave background. It has been speculated that a vast void might contribute to the amplitude of the Cold Spot from the integrated Sachs–Wolfe effect.

  10. Hot spots of mutualistic networks.

    Science.gov (United States)

    Gilarranz, Luis J; Sabatino, Malena; Aizen, Marcelo A; Bascompte, Jordi

    2015-03-01

    Incorporating interactions into a biogeographical framework may serve to understand how interactions and the services they provide are distributed in space. We begin by simulating the spatiotemporal dynamics of realistic mutualistic networks inhabiting spatial networks of habitat patches. We proceed by comparing the predicted patterns with the empirical results of a set of pollination networks in isolated hills of the Argentinian Pampas. We first find that one needs to sample up to five times as much area to record interactions as would be needed to sample the same proportion of species. Secondly, we find that peripheral patches have fewer interactions and harbour less nested networks - therefore potentially less resilient communities - compared to central patches. Our results highlight the important role played by the structure of dispersal routes on the spatial distribution of community patterns. This may help to understand the formation of biodiversity hot spots. © 2014 The Authors. Journal of Animal Ecology © 2014 British Ecological Society.

  11. Suscetibilidade aos agentes quimioterápicos de isolados de Schistosoma mansoni oriundos de pacientes tratados com oxamniquine e praziquantel e não curados

    Directory of Open Access Journals (Sweden)

    Neusa Araújo

    1996-10-01

    Full Text Available Foram estudados dez isolados de Schistosoma mansoni provenientes de pacientes residentes em Itaquara, Bahia, Brasil, tratados com oxamniquine eposteriormente com praziquantel, e ainda assim não curados. Caramujos (Biomphalaria glabrataj foram expostos a miracídios provenientes das fezes dos pacientes. Cercãrías eliminadas por estes caramujos e por moluscos coletados no peridomicílio dos pacientes foram utilizadas para infecção experimental de camundongos albinos. Os animais infectados foram tratados em dose única, via oral, com oxamniquine (25, 50 e 100 mg/kg ou praziquantel (100, 200 e 400mg/kg. Foram realizados estudos de análise e comparação de DNA de cercãrías de S. mansoni eliminadas pelos caramujos e de vermes adultos recolhidos de camundongos infectados experimentalmente com os isolados dos 10 pacientes e ainda de cercãrías de S. mansoni eliminadas por moluscos naturalmente infectados coletados em Itaquara. A cepa LE (mantida rotineiramente no laboratório foi usada como padrão de comparação da resposta aos agentes esquistossomicidas administrados. As respostas terapêuticas foram significativamente diferentes entre alguns dos isolados embora não fosse possível caracterizar nenhum como resistente. A análise dos perfis de amplificação de DNA nas cercãrías e nos vermes adultos dos isolados de S. mansoni demonstrou baixo grau de variabilidade indicando que estes são geneticamente próximos e revelando a ausência de rearranjos globais dentro dos genomas.

  12. Frequência de genes codificadores de toxinas em Staphylococcus aureus isolados de leite de tanques expansão comunitários

    Directory of Open Access Journals (Sweden)

    Atzel Candido Acosta

    Full Text Available RESUMO: A capacidade de produção de toxinas pelo Staphylococcus aureus no leite e produtos derivados está relacionado com surtos de intoxicação alimentar. Objetivou-se nesta pesquisa, estudar a ocorrência de genes que codificam para enterotoxinas estafilocócicas (sea, seb, sed, seg, seh e sei e toxinas α e β hemolítica (hla e hlb em S. aureus isolados de 53 amostras de leite de tanques expansão comunitários no Estado de Alagoas, Brasil. Foram identificados 27 isolados (50,94% como S. aureus pela amplificação do gene nuc. 13/27 isolados (48,1% foram positivos para pelo menos um gene das enterotoxinas estudadas, sendo as frequências dos genes sea 33,3%, seh 18,5%, sei 11,1% e sed 7,4%; não entanto não foram identificados os genes seb e seg nestas bactérias. Para as toxinas hemolíticas, 51,9% dos isolados portavam ambos genes (hla e hlb, sendo a frequência para o gene hla de 81,5% e para o gene hlb de 51,9%. A frequência de genes das toxinas avaliadas é alta o que constitui um risco potencial para a saúde pública em especial, as enterotoxinas por serem termoestáveis e estarem asssociados com surtos de intoxicação alimentar.

  13. Presence and Distribution of Tobacco Viruses in Serbia

    Directory of Open Access Journals (Sweden)

    Nataša Duduk

    2006-01-01

    Full Text Available Infection with a large number of plant viruses could imperil tobacco yield and quality. Tobacco is a natural host for more than 20 viruses, among which the most important and economically harmful are tobacco mosaic virus (TMV, tomato spotted wilt virus (TSWV, cucumber mosaic virus (CMV, potato virus Y (PVY, alfalfa mosaic virus (AMV, tobacco ringspot virus (TRSV, tobacco each virus (TEV and tobacco vein mottling virus (TVMV.The occurence and distribution of tobacco viruses were investigated for 4 years (2002-2005. During this period many different tobacco growing localities in Vojvodina and central Serbia were monitored and samples showing virus symptoms were collected. The collected samples were tested by DAS ELISA using polyclonal antisera, specific for the detection of PVY, TSWV, TMV, CMV, AMV and TRSV.The results obtained for the tobacco virus distribution during these four years of investigation proved the presence of four economically important viruses in our country, whose frequencies varied from year to year. In 2002, 2003 and 2004, the most frequent was TSWV(86.84%; 79% and 49.56%, respectively, while in 2005 PVY was prevalent (56.16%. All viruses detected in the samples tested were present in single or mixed infections. A corellation was established between the field symptoms on tobacco and the virus causal agents.The results obtained showed that TSWV and PVY were the most important tobacco viruses in our country, so that further research of tobacco virus diseases should be directed towards their more detailed characterization.

  14. Virus - vector relationships in the transmission of tospoviruses

    NARCIS (Netherlands)

    Wijkamp, I.

    1995-01-01

    Tomato spotted wilt virus (TSWV), member of the genus Tospovirus within the family Bunyaviridae, ranks among the top ten of economically most important plant viruses. Tospoviruses cause significant yield losses in agricultural crops such as tomato,

  15. The biology of the California spotted owl

    Science.gov (United States)

    R.J. Gutiérrez; Douglas J. Tempel; M. Zachariah Peery

    2017-01-01

    The spotted owl (Strix occidentalis) is one of the most studied raptors in the world (Lõmus 2004) because forest management throughout its range has the potential to negatively affect owl populations. Information on the California spotted owl (S. o. occidentalis) has been summarized in several literature reviews (e.g.,...

  16. Front blind spot crashes in Hong Kong.

    Science.gov (United States)

    Cheng, Yuk Ki; Wong, Koon Hung; Tao, Chi Hang; Tam, Cheok Ning; Tam, Yiu Yan; Tsang, Cheuk Nam

    2016-09-01

    In 2012-2014, our laboratory had investigated a total of 9 suspected front blind spot crashes, in which the medium and heavy goods vehicles pulled away from rest and rolled over the pedestrians, who were crossing immediately in front of the vehicles. The drivers alleged that they did not see any pedestrians through the windscreens or the front blind spot mirrors. Forensic assessment of the goods vehicles revealed the existence of front blind spot zones in 3 out of these 9 accident vehicles, which were attributed to the poor mirror adjustments or even the absence of a front blind spot mirror altogether. In view of this, a small survey was devised involving 20 randomly selected volunteers and their goods vehicles and 5 out of these vehicles had blind spots at the front. Additionally, a short questionnaire was conducted on these 20 professional lorry drivers and it was shown that most of them were not aware of the hazards of blind spots immediately in front of their vehicles, and many did not use the front blind spot mirrors properly. A simple procedure for quick measurements of the coverage of front blind spot mirrors using a coloured plastic mat with dimensional grids was also introduced and described in this paper. Copyright © 2016 Elsevier Ireland Ltd. All rights reserved.

  17. Hot Spot Removal System: System description

    Energy Technology Data Exchange (ETDEWEB)

    NONE

    1997-09-01

    Hazardous wastes contaminated with radionuclides, chemicals, and explosives exist across the Department of Energy complex and need to be remediated due to environmental concerns. Currently, an opportunity is being developed to dramatically reduce remediation costs and to assist in the acceleration of schedules associated with these wastes by deploying a Hot Spot Removal System. Removing the hot spot from the waste site will remove risk driver(s) and enable another, more cost effective process/option/remedial alternative (i.e., capping) to be applied to the remainder of the site. The Hot Spot Removal System consists of a suite of technologies that will be utilized to locate and remove source terms. Components of the system can also be used in a variety of other cleanup activities. This Hot Spot Removal System Description document presents technologies that were considered for possible inclusion in the Hot Spot Removal System, technologies made available to the Hot Spot Removal System, industrial interest in the Hot Spot Removal System`s subsystems, the schedule required for the Hot Spot Removal System, the evaluation of the relevant technologies, and the recommendations for equipment and technologies as stated in the Plan section.

  18. Rocky Mountain spotted fever in children.

    Science.gov (United States)

    Woods, Charles R

    2013-04-01

    Rocky Mountain spotted fever is typically undifferentiated from many other infections in the first few days of illness. Treatment should not be delayed pending confirmation of infection when Rocky Mountain spotted fever is suspected. Doxycycline is the drug of choice even for infants and children less than 8 years old. Copyright © 2013 Elsevier Inc. All rights reserved.

  19. 7 CFR 28.411 - Good Middling Light Spotted Color.

    Science.gov (United States)

    2010-01-01

    ... 7 Agriculture 2 2010-01-01 2010-01-01 false Good Middling Light Spotted Color. 28.411 Section 28... Light Spotted Color. Good Middling Light Spotted Color is color which in spot or color, or both, is between Good Middling Color and Good Middling Spotted Color. ...

  20. 7 CFR 28.413 - Middling Light Spotted Color.

    Science.gov (United States)

    2010-01-01

    ... 7 Agriculture 2 2010-01-01 2010-01-01 false Middling Light Spotted Color. 28.413 Section 28.413... Spotted Color. Middling Light Spotted Color is color which in spot or color, or both, is between Middling Color and Middling Spotted Color. ...

  1. 7 CFR 28.415 - Low Middling Light Spotted Color.

    Science.gov (United States)

    2010-01-01

    ... 7 Agriculture 2 2010-01-01 2010-01-01 false Low Middling Light Spotted Color. 28.415 Section 28... Spotted Color. Low Middling Light Spotted Color is color which in spot or color, or both, is between Low Middling Color and Low Middling Spotted Color. ...

  2. 7 CFR 28.412 - Strict Middling Light Spotted Color.

    Science.gov (United States)

    2010-01-01

    ... 7 Agriculture 2 2010-01-01 2010-01-01 false Strict Middling Light Spotted Color. 28.412 Section 28... Light Spotted Color. Strict Middling Light Spotted Color is color which in spot or color, or both, is between Strict Middling Color and Strict Middling Spotted Color. ...

  3. Quantitative control of Lettuce mosaic virus fitness and host defence inhibition by P1-HCPro P1-HC Pro do Lettuce mosaic virus atua de forma quantitativa na inibição da resposta de defesa do hospedeiro e adaptação viral

    Directory of Open Access Journals (Sweden)

    Renate Krause-Sakate

    2007-06-01

    Full Text Available Two Lettuce mosaic virus isolates capable of overcoming the resistance afforded by the resistance gene mo1² in lettuce, LMV-AF199 from Brazil, and LMV-E, an European isolate, were evaluated for the rapidity and severity of symptoms induced on the lettuce variety Salinas 88 (mo1². The mosaic symptoms on Salinas 88 plants inoculated with LMV-AF199 appeared 7 days post-inoculation (dpi and 15 dpi for LMV-E. The symptoms induced by LMV-AF199 in this cultivar were also more severe than those induced by LMV-E. In order to identify the region of the viral genome responsible for this phenotype, recombinant viruses were constructed between these isolates and the phenotype of each recombinant was analysed. The region encoding proteins P1 and HcPro from LMV-AF199 was associated with the increased virulence in Salinas 88.Dois isolados de Lettuce mosaic virus capazes de contornar a resistência conferida pelo gene mo1² em alface, LMV-AF199 proveniente do Brasil e LMV-E um isolado europeu, foram avaliados quanto à rapidez e à severidade dos sintomas induzidos em alface variedade Salinas 88 (mo1². Os sintomas de mosaico induzidos pelo isolado LMV-AF-199 em Salinas 88 são mais severos e aparecem aos 7 dias após a inoculação (dpi, enquanto que para o isolado LMV-E os sintomas são visíveis somente a partir dos 15 dpi. Com o intuito de identificar a região do genoma viral responsável por este fenótipo, vírus recombinantes foram construídos entre estes dois isolados, e o fenótipo avaliado quanto a rapidez e severidade dos sintomas em Salinas-88. A região codificadora para as proteínas P1 e Hc-Pro do LMV-AF199 foi associada com o aumento da virulência deste isolado em Salinas-88.

  4. Laser Spot Detection Based on Reaction Diffusion

    Directory of Open Access Journals (Sweden)

    Alejandro Vázquez-Otero

    2016-03-01

    Full Text Available Center-location of a laser spot is a problem of interest when the laser is used for processing and performing measurements. Measurement quality depends on correctly determining the location of the laser spot. Hence, improving and proposing algorithms for the correct location of the spots are fundamental issues in laser-based measurements. In this paper we introduce a Reaction Diffusion (RD system as the main computational framework for robustly finding laser spot centers. The method presented is compared with a conventional approach for locating laser spots, and the experimental results indicate that RD-based computation generates reliable and precise solutions. These results confirm the flexibility of the new computational paradigm based on RD systems for addressing problems that can be reduced to a set of geometric operations.

  5. Neptune's small dark spot (D2)

    Science.gov (United States)

    1999-01-01

    This bulls-eye view of Neptune's small dark spot (D2) was obtained by Voyager 2's narrow-angle camera. Banding surrounding the feature indicates unseen strong winds, while structures within the bright spot suggest both active upwelling of clouds and rotation about the center. A rotation rate has not yet been measured, but the V-shaped structure near the right edge of the bright area indicates that the spot rotates clockwise. Unlike the Great Red Spot on Jupiter, which rotates counterclockwise, if the D2 spot on Neptune rotates clockwise, the material will be descending in the dark oval region. The fact that infrared data will yield temperature information about the region above the clouds makes this observation especially valuable. The Voyager Mission is conducted by JPL for NASA's Office of Space Science and Applications.

  6. Utilização do antibiograma como ferramenta de tipagem fenotípica de Staphylococcus aureus isolados de manipuladores , leite cru e queijo minas frescal em laticínio de Goiás, Brasil

    Directory of Open Access Journals (Sweden)

    Maria Cláudia Dantas Porfirio Borges André

    2006-12-01

    Full Text Available O trabalho teve por objetivo isolar, identificar e caracterizar fenotipicamente utilizando o antibiograma, Staphylococcus aureus isolados de manipuladores, leite cru e queijo Minas Frescal, em um laticínio de Goiás. Durante doze meses (Março/2004 a Fevereiro/2005 140 amostras foram analisadas. As cepas foram isoladas de 75% dos manipuladores resultando em 31 (33,7% isolados a partir de 92 amostras coletadas (46 de mãos e 46 de nasofaringe. Das 24 amostras de leite cru, 18 (75,0% foram positivas, resultando em 26 isolados, com média de contagem de 1,1 x 10(5 UFC/ml. Das 24 amostras de queijo 17 (70,8% foram positivas originando 20 isolados, com média de 3,8 x 10(4 UFC/g, sendo 13 (54,2% com população acima do limite estabelecido pela legislação brasileira (10³ UFC/g. A tipagem das cepas foi feita pelo antibiograma, através do método de difusão em placas. Todos os isolados foram susceptíveis à ciprofloxacina e gentamicina. Foi observada resistência em cinco (6,5% isolados para eritromicina, 19 (24,7% para tetraciclina, um (1,3% para vancomicina, quatro (5,2% para oxacilina e 53 (68,8% para penicilina. Onze (14,3% isolados foram susceptíveis a todos os antibióticos testados e 18 (23,4% foram resistentes a mais de um antibiótico. O antibiograma permitiu a classificação das cepas em doze perfis diferentes (A-L, porém não foi uma técnica eficiente em determinar a origem da contaminação final do queijo.

  7. Natural incidence of bean viruses in the northwest of Iran

    Directory of Open Access Journals (Sweden)

    Mina Rastgou

    2017-09-01

    Full Text Available Bean is considered as one of the most important legumes around the world. Viral diseases are a major yield reducing factor in bean production. Bean samples with virus-like symptoms like severe or mild mosaic, vein banding, leaf curling, blistering and necrosis were collected from different bean fields in Urmia (Northwest of Iran during the growing seasons of 2013 and 2014. Bean common mosaic virus (BCMV, Bean common mosaic necrosis virus (BCMNV, Bean yellow mosaic virus (BYMV, Cucumber mosaic virus (CMV, Tomato spotted wilt virus (TSWV, Tomato mosaic virus (ToMV and Tomato yellow leaf curl virus (TYLCV were detected by double antibody sandwich enzyme-linked-immunosorbent assay. Mixed infection of BCMV and BCMNV were found. BCMNV was the most frequent virus in this region whereas BYMV and TYLCV were each detected just in one sample. This is the first report of BCMNV, BCMV, BYMV, TSWV, TMV and TYLCV incidence on bean in Urmia, Iran.

  8. HUBBLE FINDS NEW DARK SPOT ON NEPTUNE

    Science.gov (United States)

    2002-01-01

    NASA's Hubble Space Telescope has discovered a new great dark spot, located in the northern hemisphere of the planet Neptune. Because the planet's northern hemisphere is now tilted away from Earth, the new feature appears near the limb of the planet. The spot is a near mirror-image to a similar southern hemisphere dark spot that was discovered in 1989 by the Voyager 2 probe. In 1994, Hubble showed that the southern dark spot had disappeared. Like its predecessor, the new spot has high altitude clouds along its edge, caused by gasses that have been pushed to higher altitudes where they cool to form methane ice crystal clouds. The dark spot may be a zone of clear gas that is a window to a cloud deck lower in the atmosphere. Planetary scientists don t know how long lived this new feature might be. Hubble's high resolution will allow astronomers to follow the spot's evolution and other unexpected changes in Neptune's dynamic atmosphere. The image was taken on November 2, 1994 with Hubble's Wide Field Planetary Camera 2, when Neptune was 2.8 billion miles (4.5 billion kilometers) from Earth. Hubble can resolve features as small as 625 miles (1,000 kilometers) across in Neptune's cloud tops. Credit: H. Hammel (Massachusetts Institute of Technology) and NASA

  9. ECHO virus

    Science.gov (United States)

    ... page: //medlineplus.gov/ency/article/001340.htm ECHO virus To use the sharing features on this page, please enable JavaScript. Enteric cytopathic human orphan (ECHO) viruses are a group of viruses that can lead ...

  10. Enhancement of superoxide dismutase and catalase activity in juvenile brown shrimp, Farfantepenaeus californiensis (Holmes, 1900, fed β-1.3 glucan vitamin E, and β-carotene and infected with white spot syndrome virus Incremento de la actividad superóxido dismutasa y catalasa en juveniles de camarón café Farfantepenaeus californiensis (Holmes, 1900 alimentados con β-1,3 glucano vitamina E y β-caroteno e infectados con el virus de la mancha blanca

    Directory of Open Access Journals (Sweden)

    Rosario Pacheco

    2011-11-01

    Full Text Available The effect of dietary β-Ο-glucan, vitamin E, and β-carotene supplements in juvenile brown shrimp, Farfantepenaeus californiensis, inoculated with white spot syndrome virus (WSSV was evaluated. Groups of 30 organisms (weighing 1 ± 0.5 g were cultured in 60 L fiberglass tanks and fed daily with β-1.3-glucan (0.1%, vitamin E (0.01%, and β-carotene (0.01% for 23 days; the specimens were then inoculated with WSSV. The antioxidant activity of the enzymes superoxide dismutase (SOD and catalase (CAT were determined in the hepatopancreas and muscle at 0, 1, 6, 12, 24, and 48 h after inoculation. Shrimp fed with β-1.3-glucan, vitamin E, and β-carotene significantly increased SOD activity in the hepatopancreas and muscle at 12 and 24 h post-infection, respectively. Shrimp fed with vitamin E and β-1.3-glucan registered an increment in SOD activity from 12 to 48 h post-infection. Shrimp fed with β-carotene increased SOD activity before infection with WSSV, and shrimp fed with β-1.3-glucan and vitamin E increased CAT activity, also before infection. The CAT activity response in shrimp muscle increased with respect to the control group for all treatments tested from 1 to 6 h after inoculation with WSSV. The highest antioxidant response was registered in shrimp fed with vitamin E. Juvenile shrimp fed with vitamin E and later inoculated with WSSV registered 100% mortality at 72 h, but shrimp fed with β-Ο-glucan and β-carotene showed greater resistance to WSSV, with mortality at 144 h post-infection. This study demonstrated the capacity of juvenile Farfantepenaeus californiensis fed β-Ο-glucan, vitamin E, or β-carotene to increase the antioxidant response before and after viral infection.Se evaluó el efecto de β-1,3-glucano, vitamina E y β-caroteno en la dieta de juveniles de camarón café Farfantepenaeus californiensis inoculados con virus del síndrome de la mancha blanca (WSSV. Se colocaron grupos de 30 camarones (peso 1 ± 0,5 g en

  11. The sweet spot of a baseball bat

    Science.gov (United States)

    Cross, Rod

    1998-09-01

    The sweet spot of a baseball bat, like that of a tennis racket, can be defined either in terms of a vibration node or a centre of percussion. In order to determine how each of the sweet spots influences the "feel" of the bat, measurements were made of the impact forces transmitted to the hands. Measurements of the bat velocity, and results for a freely suspended bat, were also obtained in order to assist in the interpretation of the force waveforms. The results show that both sweet spots contribute to the formation of a sweet spot zone where the impact forces on the hands are minimised. The free bat results are also of interest since they provided particularly elegant examples of wave excitation and propagation, suitable for a student demonstration or experiment.

  12. Rocky Mountain Spotted Fever (For Parents)

    Science.gov (United States)

    ... Safety for the Whole Family Evaluate Your Child's Lyme Disease Risk Lyme Disease Lyme Disease Hey! A Tick Bit Me! Bug Bites and Stings Rocky Mountain Spotted Fever Lyme Disease Contact Us Print Resources Send to a Friend ...

  13. Scientists Spot Genes Behind Crohn's, Ulcerative Colitis

    Science.gov (United States)

    ... medlineplus.gov/news/fullstory_166957.html Scientists Spot Genes Behind Crohn's, Ulcerative Colitis Large study finds key ... Researchers say they've come closer to pinpointing genes linked with inflammatory bowel diseases such as Crohn's ...

  14. Microabrasion: a treatment option for white spots.

    Science.gov (United States)

    Souza de Barros Vasconcelos, M Q; Almeida Vieira, K; da Consolação Canuto Salgueiro, M; Almeida Alfaya, T; Santos Ferreira, C; Bussadori, S K

    2014-01-01

    The aim of the present study was to describe a clinical protocol for the treatment of white spots with the use of an abrasive material. A four-year-old patient presented with a white spot on tooth 51 and a white spot associated with a carious lesion in the cervical region of tooth 52. Treatment was planned with microabrasion and restoration of the upper right lateral incisor. Prophylaxis was first performed, followed by protection with a dental dam and the application of the abrasive material (silicon carbide and hydrochloric acid 6%). Five applications were needed to remove the spots. The restoration of the upper right lateral incisor was then performed with a resin composite. A good esthetic outcome was achieved and both the patient and her guardians were satisfied with the results. Microabrasion is a conservative treatment option that achieves satisfactory results with regard to tooth color.

  15. SPOT Controlled Image Base 10 meter

    Data.gov (United States)

    U.S. Geological Survey, Department of the Interior — SPOT Controlled Image Base 10 meter (CIB-10) is a collection of orthorectified panchromatic (grayscale) images. The data were acquired between 1986 and 1993 by the...

  16. 5 Ways to Spot a Fad Diet

    Science.gov (United States)

    ... a Change – Your Personal Plan Hot Topics Breast Exams Sexual Harassment and Sexual Bullying Prescription Drug Abuse ... 5 Ways to Spot a Fad Diet Lots of today's popular diets take advantage of our desire to drop weight quickly. Unfortunately, ...

  17. Lisch spots in neurofibromatosis type 1

    Directory of Open Access Journals (Sweden)

    Saxena Rajesh

    1991-01-01

    Full Text Available The eyes of 28 patients of Neurofibromatosis type 1 were examined. Lisch spots were present in all the patients above 20 years. Their clinical appearance is being presented as seen in Indian subjects.

  18. Bias Blind Spot: Structure, Measurement, and Consequences

    OpenAIRE

    Scopelliti, I.; Morewedge, C. K.; McCormick, E.; Min, L.; LeBrecht, S.; Kassam, K.

    2015-01-01

    People exhibit a bias blind spot: they are less likely to detect bias in themselves than in others. We report the development and validation of an instrument to measure individual differences in the propensity to exhibit the bias blind spot that is unidimensional, internally consistent, has high test-retest reliability, and is discriminated from measures of intelligence, decision making ability, and personality traits related to self-esteem, self-enhancement, and self-presentation. The scale ...

  19. Sweet spot supersymmetry and composite messengers

    OpenAIRE

    Ibe, Masahiro; Kitano, Ryuichiro

    2008-01-01

    Sweet spot supersymmetry is a phenomenologically and cosmologically perfect framework to realize a supersymmetric world at short distance. We discuss a class of dynamical models of supersymmetry breaking and its mediation whose low-energy effective description falls into this framework. Hadron fields in the dynamical models play a role of the messengers of the supersymmetry breaking. As is always true in the models of the sweet spot supersymmetry, the messenger scale is predicted to be 10^5 G...

  20. Production System of Virus-free Apple Plants Using Heat Treatment and Shoot Tip CultureShoot Tip Culture

    OpenAIRE

    Gunsup Lee; Il Sheob Shin; Kang Hee Cho; Se Hee Kim; Juhee Shin; Dae Hyun Kim; Jeong Hee Kim; Hyun Ran Kim

    2013-01-01

    In worldwide, viral diseases of apple plants has caused the serious problems like reduced production and malformation of fruits. Also, the damages of apple plants by virus and/or viroid infection (Apple chlorotic leaf spot virus, Apple stem grooving virus, Apple mosaic virus, and Apple scar skin viroid) were reported in Korea. However there is few report about the protection approach against the infection by apple viruses. Therefore, this paper introduced the experimental protocol...

  1. Susceptibilidade antimicrobiana de Campylobacter fetus subsp. venerealis isolado de bovinos Antimicrobial susceptibility of Campylobacter fetus subsp. venerealis isolated from cattle

    Directory of Open Access Journals (Sweden)

    Agueda C. Vargas

    2005-03-01

    Full Text Available A campilobacteriose venérea bovina, ocasionada principalmente pelo Campylobacter fetus subsp. fetus e Campylobacter subsp. venerealis, é transmitida através do coito ou por inseminação com sêmen contaminado. O propósito deste estudo foi determinar a susceptibilidade in vitro de isolados de C. fetus subesp. venerealis a agentes antimicrobianos comumente utilizados para o tratamento clínico e de sêmen. Foram testadas duas cepas padrão, sendo uma de C. fetus subsp. fetus e outra de C. fetus subsp. venerealis, bem como 21 amostras de isolados clínicos de C. fetus subsp. venerealis. Os testes foram realizados conforme o método de Kirby-Bauer. A amostra padrão de C. fetus subsp. fetus demonstrou-se resistente à lincomicina, penicilina e ácido nalidíxico, enquanto a de C. fetus subsp. venerealis apresentou susceptibilidade a todos antimicrobianos testados, com exceção do ácido nalidíxico. Todas as amostras de C. fetus subsp. venerealis foram susceptíveis à amicacina, ampicilina, cefalotina, estreptomicina, gentamicina, penicilina e tetraciclina. Foi observada resistência de 42,86% à lincomicina e 4,76 % a enrofloxacina, e de 100% ao ácido nalidíxico. Ainda, 4,76% apresentaram susceptibilidade intermediária à enrofloxacina, neomicina e polimixina B e 9,52% à lincomicina. Os resultados evidenciaram a sensibilidade das amostras analisadas aos antimicrobianos comumente utilizados para o tratamento clínico e do sêmen.Venereal campylobacteriosis is associated with infection of Campylobacter fetus subsp. fetus and Campylobacter fetus subsp. venerealis. The etiological agent is transmitted by natural bull breeding or artificial insemination using contaminated semen. The present study aimed to determine the in vitro susceptibility of C. fetus subsp. venerealis isolates to antimicrobial drugs generally used in clinical and semen treatment. Reference strains of C. fetus subsp. fetus and C. fetus subsp. venerealis and 21 C. fetus

  2. Halolike Phenomenon Around a Café au Lait Spot Superimposed on a Mongolian Spot.

    Science.gov (United States)

    Neri, Iria; Lambertini, Martina; Tengattini, Vera; Rivalta, Beatrice; Patrizi, Annalisa

    2017-05-01

    An 8-month-old Caucasian infant with neurofibromatosis type 1 presented with a congenital plexiform neurofibroma and multiple café au lait spots. A pale area surrounded one of the café au lait spots located on the left gluteus in the area of dermal melanocytosis. This halolike phenomenon results from the disappearance of the Mongolian spot around the café au lait spots, revealing normal pigmented skin. This sign has been described rarely in the literature and the pathogenic mechanism is unclear. © 2017 Wiley Periodicals, Inc.

  3. Propriedades probióticas in vitro de Lactobacillus spp. isolados de queijos minas artesanais da Serra da Canastra - MG

    Directory of Open Access Journals (Sweden)

    C.R.G. Andrade

    2014-10-01

    Full Text Available O objetivo deste estudo foi determinar o potencial probiótico in vitro de Lactobacillus spp. isolados de queijos minas artesanais da Serra da Canastra, considerando-se o antagonismo entre amostras isoladas frente a microrganismos indicadores, a susceptibilidade a antimicrobianos, a sensibilidade ao ácido gástrico e a sensibilidade a sais biliares. Todas as bactérias ácido-lácticas testadas apresentaram resistência ao ácido gástrico (pH 2,0 e aos sais biliares (0,3%, bem como atividade antagonista contra Escherichia coli, Staphylococcus aureus, Listeria monocytogenes, Salmonella enterica var. Typhimurium, Enterococcus faecalis e bactérias ácido-lácticas isoladas dos próprios queijos - Lactobacillus plantarum (D27 e Lactobacillus rhamnosus (B25. Todas as amostras foram sensíveis à eritromicina e tetraciclina e resistentes à ciprofloxacina, gentamicina, oxacilina, estreptomicina e vancomicina. L. plantarum (B17 apresentou melhor potencial probiótico, pois obteve resultados satisfatórios em todas as propriedades avaliadas. Mais estudos são necessários para verificar a presença e a capacidade de transmissão de genes de resistência antimicrobiana a outros microrganismos e para avaliar o potencial dos microrganismos in vivo. As bactérias selecionadas poderão ser utilizadas na elaboração de queijos em que sejam mantidos o sabor e a tradição do queijo minas artesanal do estado de Minas Gerais.

  4. Análise genotípica de isolados de Mycobacterium tuberculosis de um hospital em Lisboa, Portugal

    Directory of Open Access Journals (Sweden)

    João Perdigão

    2009-09-01

    Full Text Available Resumo: Portugal apresenta uma das taxas de incidência de tuberculose mais altas da União Europeia, tendo a Região de Saúde de Lisboa uma taxa de incidência muito acima da média nacional. O presente estudo analisa a transmissão, susceptibilidade aos antibacilares e características de uma população de estudo de um hospital central de Lisboa. Cento e trinta e dois isolados clínicos de Mycobacterium tuberculosis foram pre-viamente testados quanto à susceptibilidade aos antibacilares de primeira linha. A taxa de multirresistência encontrada foi de 3,0%, enquanto 13,6% dos isolados eram resistentes a um ou mais antibacilares de primeira linha. A serologia para o VIH estava disponível para 98 doentes, 26 (26,5% eram positivos. Os isolados foram genotipados por MIRU-VNTR e 53 (40,2% dos 132 isolados encontravam-se distribuídos por 17 clusters MIRU-VNTR diferentes de dois ou mais isolados. De todos os isolados analisados, 25,8% perten-ciam à família Lisboa. Concluímos que a transmissão de estirpes de Mycobacterium tuberculosis, resistentes e susceptíveis, está a ocorrer, com especial preocupação para as estirpes Lisboa.Rev Port Pneumol 2009; XV (5: 761-769 Abstract: Portugal has one of the highest tuberculosis notification rates of the European Union with Lisbon Health Region having an incidence rate well above the national average. The present study analyses the transmission, drug susceptibility and characteristics of a study population from a Central Lisbon’s Hospital. One hundred and thirty-two Mycobacterium tuberculosis clinical isolates were previously tested for drug susceptibility to first-line drugs. The multidrug (MDR resistance rate was found to be 3.0%, while 13.6% of the isolates were resistant to one or more first-line drugs. HIV serology was available for 98 patients, 26 (26.5% were positive. Genotyping was performed by MIRU-VNTR and 53 (40,2% out of the 132 isolates

  5. Mancha de mirotécio em algodoeiro causada por Myrothecium roridum Myrothecium leaf spot of cotton caused by Myrothecium roridum

    Directory of Open Access Journals (Sweden)

    Maurício Conrado Meyer

    2006-09-01

    Full Text Available A mancha de mirotécio causada por Myrothecium roridum Tode ex Fr. foi observada em lavouras de algodão no sul do Maranhão, causando reduções de produtividade de até 60%. Os sintomas da doença são lesões necróticas, circulares, com estruturas salientes, os esporodóquios, de distribuição irregular. Foram observadas lesões nos pecíolos, brácteas, folhas e maçãs de algodoeiro cv. Deltapine Acala 90, Fibermax 966 e Sure Grow 821. O isolamento do fungo foi realizado em meio de cultura batata-dextrose-ágar (BDA. O teste de patogenicidade foi realizado em maçãs sadias, destacadas de algodoeiro cv. Fibermax 966, no estádio vegetativo R6, previamente desinfestadas. Foram testados 13 isolados de M. roridum, oito provenientes de algodão e cinco de soja. Avaliações das estruturas fúngicas foram realizadas com auxílio de microscópio óptico equipado com um micrômetro ocular. Os isolados causaram infecções em maçãs de algodão e destacou-se como mais agressivo o MA-75, proveniente de algodão, apresentando diâmetro médio de lesão de 1,3cm, aos sete DAI e 2,7cm aos 14 DAI. Todos os isolados formaram esporodóquios dispostos concenticamente em meio BDA. Os conídios são unicelulares, hialinos a oliváceos, abundantemente produzidos em massa verde-oliva a preta. Os conídios de isolados provenientes de algodão mediram, em média, 5,1µm x 1,5µm, e os obtidos de soja, 5,8µm x 1,5µm. Estes resultados relatam a ocorrência da mancha de mirotécio, causada por M. roridum, em lavouras comerciais de algodão no Brasil.Myrothecium leaf spot caused by Myrothecium roridum Tode ex Fr. was observed on cotton fields in Maranhão State, Brazil, causing yield reduction of up to 60%. Disease symptoms are lesions with concentric necrotic rings, with salient structures (sporodochia irregularly distributed. Symptoms were observed on petioles, bracts, leaves and bolls of cotton cultivars Deltapine Acala 90, Fibermax 966 and Sure Grow 821

  6. Tissue tropism related to vector competence of Frankliniella occidentalis for tomato spotted wilt tospovirus.

    Science.gov (United States)

    Nagata, T; Inoue-Nagata, A K; Smid, H M; Goldbach, R; Peters, D

    1999-02-01

    The development of tomato spotted wilt tospovirus (TSWV) infection in the midgut and salivary glands of transmitting and non-transmitting thrips, Frankliniella occidentalis, was studied to elucidate tissue tropism and the virus pathway within the body of this vector. Immunohistological techniques used in this study showed that the midgut, foregut and salivary glands were the only organs in which virus accumulated. The first signals of infection, observed as randomly distributed fluorescent granular spots, were found in the epithelial cells of the midgut, mainly restricted to the anterior region. The virus subsequently spread to the circular and longitudinal midgut muscle tissues, a process which occurred late in the larval stage. In the adult stage, the infection occurred in the visceral muscle tissues, covering the whole midgut and foregut, and was abolished in the midgut epithelium. The infection of the salivary glands was first observed 72 h post-acquisition, and simultaneously in the ligaments connecting the midgut with these glands. The salivary glands of transmitting individuals appeared heavily or completely infected, while no or only a low level of infection was found in the glands of non-transmitting individuals. Moreover, the development of an age-dependent midgut barrier against virus infection was observed in second instar larvae and adults. The results show that the establishment of TSWV infection in the various tissues and the potential of transmission seems to be regulated by different barriers and processes related to the metamorphosis of thrips.

  7. SENSIBILIDADE “IN VITRO” DE ISOLADOS DE Colletotrichum lindemuthianum A FUNGICIDAS “IN VITRO” SENSITIVITY OF ISOLATED Colletotrichum lindemuthianum TO FUNGICIDES

    Directory of Open Access Journals (Sweden)

    Aloísio Sartorato2 Sartorato

    2007-09-01

    Full Text Available

    Avaliou-se a sensibilidade "in vitro" de isolados do fungo Colletotrichum lindemuthianum aos fungicidas tiofanato metílico + clorotalonil, tiofanato metílico, fluazinam, clorotalonil, trifloxystrobin + propiconazole, piraclostrobin, difenoconazole e fentin hidróxido. Os diâmetros das colônias, sob os diferentes princípios ativos avaliados, diferiram significativamente da testemunha. Os isolados de C. lindemuthianum apresentaram alta sensibilidade aos princípios ativos difenoconazole, piraclostrobin, trifloxistrobin + propiconazole e fluazinam. A maioria dos isolados apresentou baixa sensibilidade ao tiofanato metílico, o que pode indicar a ocorrência de resistência do fungo C. lindemuthinaum a esse princípio ativo.

    PALAVRAS-CHAVE: Antracnose; resistência a fungicidas; controle químico.

    In vitro tests were carried out to evaluate the sensitivity of Colletotrichum lindemuthianum to the fungicides thiophanate methyl + chlorothalonil, thiophanate methyl, fluazinam, chlorothalonil, trifloxystrobin + propiconazole, pyraclostrobin, difenoconazole and fentin hidroxide. The fungus colony diameter of all isolates differed significantly from control in all active ingredient (a.i. used. C. lindemuthianum isolates presented higher sensitivity to the a.i. difenoconazole, pyraclostrobin, trifloxystrobin + propiconazole and fluazinam. The majority of isolates presented low sensitivity to methyl thiophanate, which may indicate the occurrence of C. lindemuthianum resistance to this active ingredient.

    KEY-WORDS: Anthracnose; resistance to fungicides; chemical control.

  8. Patogenicidade e virulência de Toxoplasma gondii isolado de suínos de criação artesanal no sul do Brasil

    Directory of Open Access Journals (Sweden)

    Plínio Aguiar de Oliveira

    2014-12-01

    Full Text Available Estudos com Toxoplasma gondii em suínos são relevantes porque seus produtos e subprodutos fazem parte da cadeia alimentar do ser humano. As principais vias de transmissão deste agente são o carnivorismo, fecal-oral e congênita. Seis isolados de Toxoplasma gondii de suínos de criação artesanal foram avaliados quanto à patogenicidade e virulência em camundongos suíços albinos. A suspensão de taquizoítos utilizada nos testes foi obtida através da punção ou lavagem da cavidade peritoneal de camundongos que apresentaram ascite. Cada amostra foi inoculada em grupos de cinco camundongos, com inóculo de 10¹, 10², 10³, 10(4, 10(5 e 10(6 taquizoítos vivos, via intraperitoneal. Dos isolados, 50% (3/6 foram letais e causaram sinais clínicos nos camundongos. A dose mínima letal foi de 10³ taquizoítos. A morte dos animais que apresentaram infecção aguda ocorreu entre 12 e 26 dias após a inoculação. Todos os isolados da região estudada apresentam alta capacidade de formar cistos, o que pode aumentar o risco de infecção pela ingestão de tecidos dos animais infectados pelos mesmos.

  9. CARACTERIZAÇÃO FUNCIONAL E NUTRICIONAL DE UM ISOLADO PROTÉICO OBTIDO A PARTIR DE RESÍDUOS DE CAMARÃO ROSA (Farfantepenaeus paulensis

    Directory of Open Access Journals (Sweden)

    P. G. COSTA

    2008-11-01

    Full Text Available

    Os resíduos da industrialização de camarão rosa em Rio Grande, sul do Brasil, geram problemas de poluição ambiental devido ao seu descarte próximo à Laguna dos Patos. Estes resíduos apresentam vários componentes de interesse: quitina, proteína e astaxantina. O objetivo do trabalho foi obter um isolado protéico a partir de resíduos da industrialização do camarão rosa e estudar suas propriedades funcionais e nutricionais. O isolado protéico apresentou 33,2% de solubilidade em pH alcalino e a capacidade de retenção de água apresentou maior percentual (1,9g H2O/ g proteína em pH ácido (2,0 e temperatura de 50ºC, capacidade de retenção de óleo de 0,9g óleo/ g proteína; capacidade de formação de espuma de 237,1% e estabilidade da espuma de 75,7%. O isolado protéico apresentou 89,0% de proteína (b.s. e coeficiente de digestibilidade "in vivo" de 82,5%. Os aminoácidos essenciais presentes em maior concentração foram leucina (5,9%, lisina (5,0% e isoleucina (3,8%, apresentando 1,9% de metionina, indicando potencial muito bom para utilização em formulações protéicas de alimentos.

  10. 7 CFR 28.423 - Middling Spotted Color.

    Science.gov (United States)

    2010-01-01

    ... 7 Agriculture 2 2010-01-01 2010-01-01 false Middling Spotted Color. 28.423 Section 28.423... REGULATIONS COTTON CLASSING, TESTING, AND STANDARDS Standards Spotted Cotton § 28.423 Middling Spotted Color. Middling Spotted Color is color which is within the range represented by a set of samples in the custody of...

  11. Serological and Molecular Characterization of a High Temperature-recovered Virus Belonging to Tospovirus Serogroup IV

    OpenAIRE

    Hei-Ti, HSU; Peter P., UENG; Fang-Hua, CHU; Zhaohui, YE; Shyi-Don, YEH; U.S.Department of Agriculture, Agricultural Research Service; Department of Plant Pathology, National Chung-Hsing University

    2000-01-01

    A serologically and cytologically distinct gloxinia tospovirus(HT-1)previously isolated from a gloxinia plant infected with Impatiens necrotic spot virus(INSV)when propagated in a high-temperature environment was characterized. Rabbit antisera produced for INSV and Tomato spotted wilt virus(TSWV)nucleocapsids(N)failed to react with HT-1 proteins in western blot analysis. The HT-1 antibodies reacted strongly with homologous antigen but failed to react with INSV and TSWV. However, the HT-1 anti...

  12. Suscetibilidade in vitro de isolados de Sporothrix schenckii frente à terbinafina e itraconazol In vitro susceptibility of isolates of Sporothrix schenckii to terbinafine and itraconazole

    Directory of Open Access Journals (Sweden)

    Ana Raquel Mano Meinerz

    2007-02-01

    Full Text Available O estudo objetiva determinar a atividade in vitro da terbinafina e itraconazol através da técnica de microdiluição em caldo (NCCLSM27-A2 adaptado para um fungo dimórfico frente a 12 isolados de Sporothrix schenckii, sendo seis de esporotricose felina, três de esporotricose humana, um isolado de cão e dois isolados humanos provenientes do Instituto Oswaldo Cruz (IOC. O inóculo e as concentrações antifúngicas foram distribuídas em microplacas, as quais foram incubadas a 35°C por cinco dias, quando foi realizada a leitura da concentração inibitória mínima. A concentração inibitória mínima para a terbinafina variou de 0,055µg/ml a 0,109µg/ml e para o itraconazol de 0,219µg/ml a 1,75µg/ml, sendo que para ambos os fármacos as CIMs entre os isolados do IOC foi de 0,875µg/ml. O estudo demonstrou uma alta suscetibilidade do Sporothrix schenckii frente à terbinafina, necessitando mais estudos que correlacionem os testes in vitro frente ao fármaco com a resposta clínica em pacientes com esporotricose.The study objective was to determine the in vitro activity of terbinafine and itraconazole through the microdilution technique in broth (NCCLSM27-A2, adapted for dimorphic fungus, in relation to 12 isolates of Sporothrix schenckii. Six were from feline sporotrichosis, three from human sporotrichosis, one from a dog and two from human isolates originating from Instituto Oswaldo Cruz. The inoculum and antifungal concentrates were distributed on microplates that were incubated at 35°C for five days. Minimum inhibitory concentration readings were made at the end of this period. The MIC for terbinafine ranged from 0.055µg/ml to 0.109µg/ml, and the MIC for itraconazole ranged from 0.219µg/ml to 1.75µg/ml. For both drugs, the MIC from the isolates from IOC was 0.875µg/ml. The present study demonstrates the high susceptibility of Sporothrix schenckii to terbinafine. Further studies to correlate the in vitro susceptibility tests with

  13. Fatores associados com a resistência a Ciprofloxacina e Levofloxacina em Bacilos Gram-Negativos isolados de infecções do trato urinário

    Directory of Open Access Journals (Sweden)

    Lisiane Martins Volcão

    2016-01-01

    Full Text Available Justificativa e objetivos: As fluoroquinolonas representam uma classe de antimicrobianos que é frequentemente prescrita no tratamento de infecções do trato urinário (ITU, tanto de origem hospitalar como originárias da comunidade. Este estudo tem como objetivo,determinar a frequência de resistência à ciprofloxacina e levofloxacina, além de seus fatores associados em bacilos Gram negativos (BGN isolados de pacientes com ITU em um hospital no Sul do Brasil. Métodos: foi realizado um estudo transversal e analítico, baseado em casos de infecções do trato urinário causadas por bactérias gram-negativas provenientes de pacientes atendidos no Hospital Universitário Dr. Miguel Riet Correa Jr. em Rio Grande/RS, de agosto de 2012 a julho 2013. Foram analisadas variáveis independentes como idade, sexo, origem da infecção e características dos isolados clínicos bacterianos. Resultados:Dos 562 BGN isolados e analisados, foi encontrado uma frequência de resistência à ciprofloxacina e à levofloxacina de 25,5 e 23,3%, respectivamente, 62,6% de origem comunitária e 59% de origem hospitalar. Os fatores de risco associados à resistência a ciprofloxacina e levofloxacina, foram pacientes do gênero masculino, infecções adquiridas no ambiente hospitalar, commaior tempo de internação, e apresença de β-lactamases de espectro estendido (ESBL nos isolados clínicos. Conclusões:Os resultados mostraram uma forte associação da resistência bacteriana de BGN com a permanência no ambiente hospitalar e a presença de ESBL. A fim de controlar a resistência antimicrobiana e a redução nos custos em saúde pública torna-se necessário que os hospitais tenham uma forte política de vigilância contínua do uso e da resistência de antibióticos.

  14. DIVERSIDADE CULTURAL, MORFOLÓGICA E GENÉTICA DE DIAZOTRÓFICOS ISOLADOS DE NÓDULOS DE BRACATINGA1

    Directory of Open Access Journals (Sweden)

    Natalia Carolina Moraes Ehrhardt-Brocardo

    2015-10-01

    Full Text Available RESUMO O nitrogênio é um dos macronutrientes essenciais aos seres vivos, o que o torna um dos fatores limitantes para o crescimento vegetal. Apenas uma parcela dos procariotos, os diazotróficos, possui a capacidade de reduzir o nitrogênio atmosférico para outras formas disponíveis às plantas. A bracatinga é uma espécie arbórea de importância econômica capaz de estabelecer simbiose mutualística com organismos diazotróficos, sendo os seus principais usos: produção de energia, madeira, forragem, indústria química de produtos naturais, apicultura, paisagismo e sombreamento e recuperação de áreas degradadas e zonas ripárias. Este estudo objetivou estabelecer relação entre a diversidade cultural e morfológica e a variabilidade genética dos isolados nodulantes em bracatinga de diferentes condições edafoclimáticas. Coletaram-se nódulos radiculares retirados ao acaso em sete áreas compreendidas entre o Vale do Itajaí, Planalto Sul e Meio-Oeste do Estado de Santa Catarina. Foi observada ampla diversidade cultural entre os diazotróficos presentes, havendo predomínio de isolados de rápido crescimento, de colônia com coloração branca leitosa, formato circular, borda lisa e superfície mucoide. Entre os parâmetros avaliados, a transparência da colônia, a produção de muco e a alteração do pH foram considerados relevantes para a diferenciação dos isolados. A caracterização taxonômica dos isolados foi realizada por comparação dos fragmentos sequenciados, sendo as espécies isoladas deste conjunto amostral pertencentes aos gêneros Burkholderia, Pantoea, Pseudomonas e Rhizobium.

  15. Micropartículas produzidas por gelificação iônica recobertas com gelatina de peixe e isolado proteico de soja

    OpenAIRE

    Bianca Souza da Costa

    2014-01-01

    Resumo: O objetivo deste trabalho foi produzir partículas de pectina e alginato por gelificação iônica, com posterior recobrimento com isolado proteico de soja (IPS), gelatina de pele de tilápia (GPT) e com a mistura dessas duas proteínas (IPS:GPT), avaliando suas características físico-químicas e seu comportamento frente a diferentes tratamentos. No estudo preliminar foram analisadas as condições que promovessem a carga elétrica livre que otimizassem a interação entre os polissacarídeos e pr...

  16. Are hot-spots occluded from water?

    Science.gov (United States)

    Moreira, Irina Sousa; Ramos, Rui Miguel; Martins, Joao Miguel; Fernandes, Pedro Alexandrino; Ramos, Maria João

    2014-01-01

    Protein-protein interactions are the basis of many biological processes and are governed by focused regions with high binding affinities, the warm- and hot-spots. It was proposed that these regions are surrounded by areas with higher packing density leading to solvent exclusion around them - "the O-ring theory." This important inference still lacks sufficient demonstration. We have used Molecular Dynamics (MD) simulations to investigate the validity of the O-ring theory in the context of the conformational flexibility of the proteins, which is critical for function, in general, and for interaction with water, in particular. The MD results were analyzed for a variety of solvent-accessible surface area (SASA) features, radial distribution functions (RDFs), protein-water distances, and water residence times. The measurement of the average solvent-accessible surface area features for the warm- and hot-spots and the null-spots, as well as data for corresponding RDFs, identify distinct properties for these two sets of residues. Warm- and hot-spots are found to be occluded from the solvent. However, it has to be borne in mind that water-mediated interactions have significant power to construct an extensive and strongly bonded interface. We observed that warm- and hot-spots tend to form hydrogen bond (H-bond) networks with water molecules that have an occupancy around 90%. This study provides strong evidence in support of the O-ring theory and the results show that hot-spots are indeed protected from the bulk solvent. Nevertheless, the warm- and hot-spots still make water-mediated contacts, which are also important for protein-protein binding.

  17. Applicability of SPOT for forest management

    Science.gov (United States)

    Jaakkola, S. P.

    The recent applications of satellite remote sensing for forest management are reviewed mainly on the basis of European experience. The review is illustrated using results of the applied research during the first two years of SPOT-1 in orbit. The potential and limitations of satellite imagery, specific to SPOT, are discussed from the forestry point of view. Research results concerning forest cover type mapping, forest inventory and change monitoring - including the European forest decline - are reported. An evaluation of digital SPOT-data for forest inventory purposes in northern Sweden is reported in more detail. The original SPOT-data was preprocessed, integrated (PA + XS-bands) and filtered for texture extraction. A texture based algorithm was tested for finding logging roads. The timber stand delineation method developed combines image segmentation, classification of the segments and the final delineation of stands using an expert system. The results show that (1) the computer-aided forest mapping leads to acceptable forest management units, and (2) the manual stand delineation using digitally enhanced SPOT-data (color prints) coincides very well with the actual forest maps used for comparison. Finally, the potential applications of satellite remote sensing for forestry in the 1990's are outlined.

  18. Spots on AG Virginis - Paradigm or panacea?

    Science.gov (United States)

    Bell, S. A.; Rainger, P. P.; Hilditch, R. W.

    1990-12-01

    New photometric and spectroscopic observations of the eclipsing binary AG Vir are presented. Medium-resolution spectroscopy has allowed the measurement of velocities for the secondary component for the first time. The V light curve shows many of the features seen in previous studies of this system. A full analysis of the spectroscopic and photometric data has been made which suggests that the system is either in a marginal state of conatact or a deep-contact configuration depending on the type of spot model invoked. AG Vir constitutes an excellent example of the expected manifestations of spot activity on a light curve. It also demonstrates the ease with which the spot phenomenon can be invoked to explain the appearance of a light curve and to provide conflicting results. This study shows the necessity of a more thorough investigation of this system using Doppler-imaging techniques and simultaneous infrared and optical photometry.

  19. Spots on AG Virginis - paradigm or panacea

    Energy Technology Data Exchange (ETDEWEB)

    Bell, S.A.; Rainger, P.P.; Hilditch, R.W. (Saint Andrews Univ. (UK). Observatory)

    1990-12-15

    New photometric and spectroscopic observations of the eclipsing binary AG Vir are presented. Medium-resolution spectroscopy has allowed the measurement of velocities for the secondary component for the first time. The V light curve shows many of the features seen in previous studies of this system. A full analysis of the spectroscopic and photometric data has been made which suggests that the system is either in a marginal state of contact or a deep-contact configuration depending on the type of spot model invoked. AG Vir constitutes an excellent example of the expected manifestations of spot activity on a light curve. It also demonstrates the ease with which the spot phenomenon can be invoked to explain the appearance of a light curve and to provide conflicting results. (author).

  20. [Rocky Mountain spotted fever in Brazil].

    Science.gov (United States)

    del Sá DelFiol, Fernando; Junqueira, Fábio Miranda; da Rocha, Maria Carolina Pereira; de Toledo, Maria Inês; Filho, Silvio Barberato

    2010-06-01

    Although the number of confirmed cases of spotted fever has been declining in Brazil since 2005, the mortality rate (20% to 30%) is still high in comparison to other countries. This high mortality rate is closely related to the difficulty in making the diagnosis and starting the correct treatment. Only two groups of antibiotics have proven clinical effectiveness against spotted fever: chloramphenicol and tetracyclines. Until recently, the use of tetracyclines was restricted to adults because of the associated bone and tooth changes in children. Recently, however, the American Academy of Pediatrics and various researchers have recommended the use of doxycycline in children. In more severe cases, chloramphenicol injections are often preferred in Brazil because of the lack of experience with injectable tetracycline. Since early diagnosis and the adequate drug treatment are key to a good prognosis, health care professionals must be better prepared to recognize and treat spotted fever.

  1. Estresse Isolado ou Associado ao Etanol Libera Prostanóides em Aorta de Ratos via ?2-Adrenoceptores

    Directory of Open Access Journals (Sweden)

    Rafaela de Fátima Ferreira Baptista

    2014-04-01

    Full Text Available Fundamento: Estresse e etanol são ambos, independentemente, importantes fatores de risco cardiovascular. Objetivo: avaliar o risco cardiovascular do consumo de etanol e exposição ao estresse, isolados e em associação, em ratos machos adultos. Métodos: Os ratos foram separados em quatro grupos: controle, etanol (20% na água de beber durante seis semanas, estresse (imobilização 1h dia/5 dias por semana/ 6 semanas e estresse/etanol. As curvas de concentração-resposta à noradrenalina - na ausência e na presença de ioimbina, L-NAME ou indometacina - ou fenilefrina foram determinadas em aortas torácicas com e sem endotélio. EC50 e resposta máxima (n = 8-12 foram comparadas através de ANOVA de dois fatores (two-way / método de Bonferroni. Resultados: Estresse ou estresse em associação com o consumo de etanol aumentaram as respostas máximas de noradrenalina em aortas intactas. Essa hiper-reatividade foi eliminada pela remoção do endotélio, ou pela presença da indometacina ou ioimbina, mas não foi alterada pela presença de L-NAME. Enquanto isso, o consumo de etanol não alterou a reatividade à noradrenalina. As respostas da fenilefrina em aortas com e sem endotélio também permaneceram inalteradas independentemente do protocolo. Conclusão: O estresse crônico aumentou as respostas aórticas dos ratos à noradrenalina. Esse efeito é dependente do endotélio vascular e envolve a liberação de prostanóides vasoconstritores através da estimulação de α-2 adrenoceptores endoteliais. Além disso, o consumo crônico de etanol pareceu não influenciar as respostas de noradrenalina em aorta de rato, nem modificar o aumento de tais respostas observadas em consequência da exposição ao estresse.

  2. Matching Two-dimensional Gel Electrophoresis' Spots

    DEFF Research Database (Denmark)

    Dos Anjos, António; AL-Tam, Faroq; Shahbazkia, Hamid Reza

    2012-01-01

    This paper describes an approach for matching Two-Dimensional Electrophoresis (2-DE) gels' spots, involving the use of image registration. The number of false positive matches produced by the proposed approach is small, when compared to academic and commercial state-of-the-art approaches. This ar......This paper describes an approach for matching Two-Dimensional Electrophoresis (2-DE) gels' spots, involving the use of image registration. The number of false positive matches produced by the proposed approach is small, when compared to academic and commercial state-of-the-art approaches...

  3. Chikungunya virus

    Science.gov (United States)

    Chikungunya virus infection; Chikungunya ... Where Chikungunya is Found Before 2013, the virus was found in Africa, Asia, Europe, and the Indian and Pacific oceans. In late 2013, outbreaks occurred for the first time in the ...

  4. Zika Virus

    Science.gov (United States)

    ... through blood transfusions. There have been outbreaks of Zika virus in the United States, Africa, Southeast Asia, the ... not travel to areas where there is a Zika virus outbreak. If you do decide to travel, first ...

  5. Chikungunya Virus

    Science.gov (United States)

    ... Gaines, PhD, MPH, MA, CHES Differentiating Chikungunya From Dengue: A Clinical Challenge For Travelers CDC Travelers' Health Chikungunya Virus Home Prevention Transmission Symptoms & Treatment Geographic Distribution Chikungunya virus in the United States ...

  6. Zika Virus

    Science.gov (United States)

    ... Funding CDC Activities For Healthcare Providers Clinical Evaluation & Disease Sexual Transmission HIV Infection & Zika Virus Testing for Zika Test Specimens – At Time of Birth Diagnostic Tests Understanding Zika Virus Test Results ...

  7. Hot-spot tectonics on Io

    Science.gov (United States)

    Mcewen, A. S.

    1985-01-01

    The thesis is that extensional tectonics and low-angle detachment faults probably occur on Io in association with the hot spots. These processes may occur on a much shorter timescale on Ion than on Earth, so that Io could be a natural laboratory for the study of thermotectonics. Furthermore, studies of heat and detachment in crustal extension on Earth and the other terresrial planets (especially Venus and Mars) may provide analogs to processes on Io. The geology of Io is dominated by volcanism and hot spots, most likely the result of tidal heating. Hot spots cover 1 to 2% of Io's surface, radiating at temperatures typically from 200 to 400 K, and occasionally up to 700K. Heat loss from the largest hot spots on Io, such as Loki Patera, is about 300 times the heat loss from Yellowstone, so a tremendous quantity of energy is available for volcanic and tectonic work. Active volcanism on Io results in a resurfacing rate as high as 10 cm per year, yet many structural features are apparent on the surface. Therefore, the tectonics must be highly active.

  8. Barley seed proteomics from spots to structures

    DEFF Research Database (Denmark)

    Finnie, Christine; Svensson, Birte

    2009-01-01

    grain filling, maturation, germination and radicle elongation. Cultivar comparisons and genetic mapping of polymorphic protein spots in doubled haploid populations provide a means to link the genome to the proteome and identify proteins that can influence grain quality. Many proteins appear in multiple...

  9. Easy Demonstration of the Poisson Spot

    Science.gov (United States)

    Gluck, Paul

    2010-01-01

    Many physics teachers have a set of slides of single, double and multiple slits to show their students the phenomena of interference and diffraction. Thomas Young's historic experiments with double slits were indeed a milestone in proving the wave nature of light. But another experiment, namely the Poisson spot, was also important historically and…

  10. WORD LEXICON REDUCTION BY CHARACTER SPOTTING

    NARCIS (Netherlands)

    Guilevic, D.; Nishiwaki, D.; Yamada, K.

    2004-01-01

    We describe a system, currently under development, to dynamically reduce a lexicon of city names, making use exclusively of the information found in a word image. Isolated characters are `spotted\\\\\\' within the word. The recognition results on those isolated characters are then used to initialize a

  11. Triggered tremor sweet spots in Alaska

    Science.gov (United States)

    Gomberg, Joan; Prejean, Stephanie

    2013-01-01

    To better understand what controls fault slip along plate boundaries, we have exploited the abundance of seismic and geodetic data available from the richly varied tectonic environments composing Alaska. A search for tremor triggered by 11 large earthquakes throughout all of seismically monitored Alaska reveals two tremor “sweet spots”—regions where large-amplitude seismic waves repeatedly triggered tremor between 2006 and 2012. The two sweet spots locate in very different tectonic environments—one just trenchward and between the Aleutian islands of Unalaska and Akutan and the other in central mainland Alaska. The Unalaska/Akutan spot corroborates previous evidence that the region is ripe for tremor, perhaps because it is located where plate-interface frictional properties transition between stick-slip and stably sliding in both the dip direction and laterally. The mainland sweet spot coincides with a region of complex and uncertain plate interactions, and where no slow slip events or major crustal faults have been noted previously. Analyses showed that larger triggering wave amplitudes, and perhaps lower frequencies (probability of triggering tremor. However, neither the maximum amplitude in the time domain or in a particular frequency band, nor the geometric relationship of the wavefield to the tremor source faults alone ensures a high probability of triggering. Triggered tremor at the two sweet spots also does not occur during slow slip events visually detectable in GPS data, although slow slip below the detection threshold may have facilitated tremor triggering.

  12. Dramatic Change in Jupiter's Great Red Spot

    Science.gov (United States)

    Simon, A. A.; Wong, M. H.; Rogers, J. H.; Orton, G. S.; de Pater, I.; Asay-Davis, X.; Carlson, R. W.; Marcus, P. S.

    2015-01-01

    Jupiter's Great Red Spot (GRS) is one of its most distinct and enduring features, having been continuously observed since the 1800's. It currently spans the smallest latitude and longitude size ever recorded. Here we show analyses of 2014 Hubble spectral imaging data to study the color, structure and internal dynamics of this long-live storm.

  13. Rocky Mountain spotted fever in dogs, Brazil.

    Science.gov (United States)

    Labruna, Marcelo B; Kamakura, Orson; Moraes-Filho, Jonas; Horta, Mauricio C; Pacheco, Richard C

    2009-03-01

    Clinical illness caused by Rickettsia rickettsii in dogs has been reported solely in the United States. We report 2 natural clinical cases of Rocky Mountain spotted fever in dogs in Brazil. Each case was confirmed by seroconversion and molecular analysis and resolved after doxycycline therapy.

  14. The sweet spots in human communication.

    Science.gov (United States)

    Salem, Philip

    2011-07-01

    In baseball, the sweet spot is a special place on a bat where the batter can hit the ball with the most power. It is the place where the performances of the batter and pitcher collide with maximum effect. It is the place where the dynamic tension between opponents leads to transformation. The dynamic tension in all living systems is between similarity and difference. Chaos and complexity scholars recognized this tension as amounts of information. When the amounts of information were high, but not too high, the system moved to the edge of chaos, to the complexity regime, to strange attractors, or to chaos, depending on the model. The sweet spot is that range of relative variety, just the proper mix of similarity and difference, leading to transformation. This essay contains a model of human communication as an emergent social process with its own sweet spots. The essay also includes a description of current literature highlighting tensions between similarity and difference, and there is an exploration of the potential to move from one basin of attraction to another. The primary constraints on finding communication sweet spots are paradigmatic - adopting a process orientation, discovering the proper parameters, bracketing sequences to define initial conditions, and understanding the strengths and weaknesses of various modeling techniques.

  15. Measuring microfocal spots using digital radiography

    Energy Technology Data Exchange (ETDEWEB)

    Fry, David A [Los Alamos National Laboratory; Ewert, Uwe [BAM

    2009-01-01

    Measurement of microfocus spot size can be important for several reasons: (1) Quality assurance during manufacture of microfocus tubes; (2) Tracking performance and stability of microfocus tubes; (3) Determining magnification is especially important for digital radiography where the native spatial resolution of the digital system is not adequate for the application; and (4) Knowledge of unsharpness from the focal spot alone. The European Standard EN 12543-5 is based on a simple geometrical method of calculating focal spot size from unsharpness of high magnification film radiographs. The following equations are used for the focal spot size measurement: By similar triangles the following equations are presupposed: f/a = U/b and M = (a+b)/a. These equations can be combined to yield the well known expression: U = f(M - 1). Solving for f, f = U/(M-1). Therefore, the focal spot size, f, can be calculated by measuring the radiographic unsharpness and magnification of a known object. This is the basis for these tests. The European standard actually uses one-half of the unsharpness (which are then added together) from both sides of the object to avoid additional unsharpness contributions due to edge transmission unsharpness of the round test object (the outside of the object is measured). So the equation becomes f = (1/2 U{sub 1} + 1/2 U{sub 2})/(M-1). In practice 1/2 U is measured from the 50% to the 90% signal points on the transition profile from ''black'' to ''white,'' (positive image) or attenuated to unattenuated portion of the image. The 50% to 90% points are chosen as a best fit to an assumed Gaussian radiation distribution from the focal spot and to avoid edge transmission effects. 1/2 U{sub 1} + 1/2 U{sub 2} corresponds about to the full width at half height of a Gaussian focal spot. A highly absorbing material (Tungsten, Tungsten Alloy, or Platinum) is used for the object. Either wires or a sphere are used as the object to

  16. Ocorrência de genes tad associados à formação de biofilme em isolados de Pasteurella multocida de pulmões de suínos com pneumonia

    Directory of Open Access Journals (Sweden)

    Danny Franciele da S.D. Moraes

    2014-12-01

    Full Text Available Os atuais sistemas de criação intensiva de suínos aumentam a pressão de seleção microbiana propiciando a disseminação de doenças respiratórias. A bactéria Pasteurella multocida é associada a diversas patologias respiratórias em animais submetidos a esse tipo de criação, causando grandes perdas econômicas. A formação de biofilme foi descrita in vitro em P. multocida e fatores analisados indicaram a facilitação na colonização dos tecidos, aumentando a resistência às defesas do hospedeiro e aos antibióticos. Os objetivos deste trabalho foram analisar a ocorrência de P. multocida em pneumonias de suínos e na microbiota de pulmões sem lesão e a ocorrência dos genes do lócus tad nestes isolados. Foram analisados 70 isolados de P. multocida de pulmões, sendo sessenta e sete com lesão e três sem lesão. Isolados do sorotipo A ocorreram principalmente em pulmões com lesões (85,71%, enquanto em pulmões sem lesão observou-se somente o sorotipo D. Os genes tadA, tadB, tadC, tadD, tadE tadF e tadG estavam presentes em 89,55% dos isolados de pulmões com lesões. Os genes tadA, tadB e tadC estavam presentes em todos os isolados de pulmões sem lesão, porém os genes tadD, tadE, tadF e tadG estavam presentes em 0%, 33,3%, 33,3% e 66,6%, dos isolados sem lesão, respectivamente. Neste trabalho observou-se a associação da ocorrência dos genes tadD, tadE e tadF em isolados de P. multocida e a presença de lesões em pulmões.

  17. Genome transcription/translation of segmented, negative-strand RNA viruses

    NARCIS (Netherlands)

    Geerts-Dimitriadou, C.

    2011-01-01

    The requirements for alignment of capped RNA leader sequences along the viral genome during influenza transcription initiation (“cap-snatching”) have long been an enigma. Previous work on Tomato spotted wilt virus (TSWV) transcription initiation has revealed that this virus displays a

  18. Mopeia Virus-related Arenavirus in Natal Multimammate Mice, Morogoro, Tanzania

    DEFF Research Database (Denmark)

    Günther, Stephan; Hoofd, Guy; Charrel, Remi

    2009-01-01

    A serosurvey involving 2,520 small mammals from Tanzania identified a hot spot of arenavirus circulation in Morogoro. Molecular screening detected a new arenavirus in Natal multimammate mice (Mastomys natalensis), Morogoro virus, related to Mopeia virus. Only a small percentage of mice carry Moro...

  19. Infection of Alstroemeria Plants with Tomato yellow ring virus in Iran

    NARCIS (Netherlands)

    Beikzadeh, N.; Bayat, H.; Jafarpour, B.; Rohani, H.; Peters, D.; Hassani-Mehraban, A.

    2012-01-01

    Alstroemeria cv. Ovation plants with virus-like necrotic spots and streaks on leaves and petals were observed in greenhouses in Khorasan Razavi (Mashhad) and Markazi (Mahallat) provinces, Iran. Samples with virus-like symptoms reacted positively in enzyme-linked immunosorbent assay with a polyclonal

  20. Tolerância de estirpes e isolados de Bradyrhizobium e de Azorhizobium a zinco, cádmio e cobre "in vitro"

    Directory of Open Access Journals (Sweden)

    I. C. B. Trannin

    2001-06-01

    Full Text Available Com o objetivo de avaliar a tolerância a metais pesados "in vitro" de estirpes inoculantes (I, isolados de solos contaminados com metais (ISC e de solos não contaminados (ISNC de Bradyrhizobium, simbiontes de Enterolobium contortisiliquum (tamboril e de Acacia mangium (acácia e de Azorhizobium, simbiontes de Sesbania virgata (sesbânia, foram realizados dois experimentos. No primeiro, dez estirpes e, ou, isolados para cada espécie vegetal foram testados em meio YMA modificado por adição de tampões biológicos (HEPES e MES, suplementado com diferentes concentrações de Cu, Cd e Zn. Cobre e Cd foram testados em concentrações de 0 a 40 mg L-1, para ambos os gêneros, enquanto Zn variou de 0 a 1.000 mg L-1, para Bradyrhizobium, e de 0 a 500 mg L-1, para Azorhizobium. O crescimento de rizóbio nas diferentes concentrações de metais foi avaliado com atribuição de valores para os padrões observados (0 a 5. Os ISC de ambos os gêneros foram mais tolerantes, mas Bradyrhizobium tolerou Zn (800 mg L-1 até duas vezes e Cu (40 mg L-1 até oito vezes mais que Azorhizobium. No segundo experimento, estirpes e isolados tolerantes (T, sensíveis (S e de tolerância média (TM a metais selecionados em meio YMA modificado foram estudados em soluções aquosas com diferentes concentrações de Cu (0 a 0,01 mg L-1, Cd e Zn (0 a 1,0 mg L-1. A avaliação do número de células viáveis em soluções de metais foi feita por contagem das unidades formadoras de colônia em 0, 24, 48, 72 e 96 h de incubação, pelo método das diluições sucessivas e inoculação em YMA. Embora as soluções de metais tenham sido mais discriminatórias quanto a tolerância a metais que o meio YMA, estes dois métodos mostraram que: (a Azorhizobium foi mais sensível que Bradyrhizobium, (b os ISC de ambos os gêneros foram mais tolerantes do que os ISNC e (c a ordem de toxidez dos metais foi Cu > Cd > Zn.

  1. Bioatividade de óleos essenciais no controle de Botrytis cinerea isolado de morangueiro Essential oils bioactivity in strawberry grey mould control

    Directory of Open Access Journals (Sweden)

    E.R. Lorenzetti

    2011-01-01

    Full Text Available Objetivou-se avaliar o uso de óleos essenciais sobre isolados de Botrytis cinerea, causador do mofo cinzento em morangueiro. Foram testados óleos essenciais de capim-limão, palmarosa, citronela, cravo, canela, menta, lavanda, tangerina, eucalipto, melaleuca, alecrim e laranja, todos estes analisados em cromatógrafo a gás acoplado a detector de massas, para identificação dos principais componentes dos óleos. Foram avaliados o crescimento micelial, produção e germinação de conídios de B. cinerea, com a incorporação do óleo no meio de cultura. Realizou-se ainda uma avaliação de voláteis e a eficiência de óleos em isolado resistente a fungicida. Para cada teste, diferentes óleos apresentaram eficiência, contudo capim limão, palmarosa, canela e menta demonstraram os melhores efeitos em todos os testes realizados. Todos os tratamentos a base de óleos demonstraram efeito semelhante a um fungicida recomendado para a cultura, a base de tiofanato metílico. Dois tratamentos mostraram-se efetivos no caso de isolado resistente (óleo de capim limão e de canela. Óleos essenciais mostram-se como opção promissora para o desenvolvimento de possíveis produtos fitossanitários para o manejo de doenças em plantas.This study aimed evaluates essential oils in Botrytis cinerea isolates growth, which causes gray mould on strawberry. Were tested essential oils of lemon grass, palmrose, citronella, clove, cinnamon, mint, lavender, tangerine, eucalyptus, tea tree, rosemary and orange. The oils were analyzed in gas chromatograph attached to mass detector for identifying the mainly oils components. Were evaluated mycelial growth, conidia production and conidia germination of B. cinerea, with oil incorporation in culture medium. Were conducted an evaluation of oils volatile components and the efficiency of oils in fungicide-resistant isolate. For each test, different oils showed efficiency, however lemongrass, palmarosa, cinnamon and mint

  2. Termorresistência de fungos filamentosos isolados de néctares de frutas envasados assepticamente Thermoresistance of filamentous fungi isolated from asceptically packaged fruit nectars

    Directory of Open Access Journals (Sweden)

    Elisa Helena da Rocha Ferreira

    2011-09-01

    Full Text Available O presente estudo teve como objetivo caracterizar a termorresistência dos fungos mais termorresistentes, isolados de néctares de maracujá e abacaxi comerciais pasteurizados, bem como verificar a produção de patulina por esses fungos, quando inoculados em tais néctares. Os fungos mais termorresistentes isolados foram o Byssochlamys fulva em néctar de maracujá e o Byssochlamys nivea em néctar de abacaxi. O B. nivea, em néctar de abacaxi, mostrou-se mais termorresistente, com base nos valores de D98°C e Z, os quais resultaram em 27 min e 5,4 °C, respectivamente, em comparação com 13,6 min e 5,5 °C encontrados para B. fulva, em néctar de maracujá. A pasteurização comumente aplicada pela indústria não se mostrou suficiente para inativação dos fungos isolados. Não houve produção de patulina quando se inocularam os fungos diretamente nos néctares estudados.This study aimed at selecting and characterising the thermal resistance of thermally resistant moulds isolated from commercial pasteurised passion fruit and pineapple nectars. It also aimed to verify patulin production by such moulds following their inoculation into the products. The most thermally resistant moulds isolated from passion fruit and pineapple nectars were Byssochlamys fulva and Byssochlamys nivea, repectively. B. nivea showed the highest thermal resistance in pineapple nectar based on the D98 °C and Z-values of 27.0 min and 5.4 °C, respectively, as compared to the same parameters found for B. fulva in passion fruit nectar of 13.6 min and 5.5 °C. The pasteurisation commonly applied by industry was shown to be inadequate to inactivate the isolated moulds. No patulin production was found when the isolated moulds were inoculated directly into the nectars.

  3. In vitro isolation and molecular characterization of an Ehrlichia canis strain from São Paulo, Brazil Isolamento e caracterização molecular de um isolado de Ehrlichia canis

    Directory of Open Access Journals (Sweden)

    Daniel M. Aguiar

    2008-09-01

    Full Text Available An Ehrlichia canis isolate was obtained from an naturally infected dog exhibiting clinical signs of ehrlichiosis in São Paulo Municipality, state of São Paulo, Brazil. The isolate was characterized by PCR and DNA sequencing of portions of the ehrlichial genes dsb, 16SrRNA, and p28. Partial dsb and 16S rRNA sequences were identical to three and five other E. canis strains, respectively, from different countries and continents (including North America, Africa, Asia and Europe. Conversely, the p28 partial sequence for this E. canis (São Paulo differed by 1, 2, and 2 nucleotides from the corresponding sequences of the E. canis strains Jake (from USA, Oklahoma (USA, and VHE (Venezuela, respectively. The results in this study indicate that E. canis is the only recognized Ehrlichia species infecting dogs in Brazil.Foi obtido um isolado de Ehrlichia canis a partir de um cão naturalmente infectado com sinais clínicos de erliquiose, oriundo do município de São Paulo, SP, Brasil. O isolado foi caracterizado molecularmente pela PCR e seqüenciamento de porções dos genes dsb,16S rRNA, e p28. A seqüência parcial dos genes dsb e 16Sr RNA apresentaram-se idênticas a três e cinco seqüências respectivamente, de E. canis provenientes de diferentes países e continentes (América do Norte, África, Ásia e Europa. Contrariamente, a seqüência parcial do gene p28 do isolado São Paulo diferiu em um nucleotídeo do isolado Jake (EUA e dois nucleotídeos dos isolados Oklahoma (EUA e VHE (Venezuelan Human Ehrlichia Venezuela. Atualmente, a E. canis é a única espécie de Ehrlichia que acomete cães no Brasil.

  4. Pepino mosaic virus and Tomato chlorosis virus causing mixed infection in protected tomato crops in Sicily

    Directory of Open Access Journals (Sweden)

    SALVATORE DAVINO

    2008-07-01

    Full Text Available An unusual virus-like yellow leaf disorder associated with fruit marbling was observed during the winter of 2005 in some greenhouse tomato crops in the province of Ragusa Sicily (Southern Italy. Leaf samples from 250 symptomatic tomato plants were serologically tested by DAS-ELISA technique for 5 viruses: Tomato spotted wilt virus (TSWV, Impatiens necrotic spot virus (INSV, Tobacco mosaic virus (TMV, Cucumber mosaic virus (CMV and Pepino mosaic virus (PepMV. PepMV was detected in 215 of the samples. The virus was mechanically transmitted to cucumber, wild metel, wild tobacco and ‘Rio Grande’ tomato. The experimental host range of PepMV-Ragusa differed from that of the PepMV found in Sardinia in 2001, which infected ‘Camone’ tomato. By applying RT-PCR to 25 PepMV-infected tomato plants, the expected 844 bp DNA fragment for PepMV and the expected 439 bp DNA fragment for Tomato chlororis virus (ToCV were obtained from all the samples tested. Sequences of the obtained amplicons were used to study the phylogenetic relationships of the viruses with isolates from other countries. Nucleotide sequence alignments showed that the sequence CP-PepMV-Ragusa (Genbank acc. No. DQ 517884 were 99% homologous with both US2 and Spain-Murcia isolates, while those of ToCV-Ragusa (Genbank acc. No. DQ517885 isolate HSP70, were 99% homologous with the Florida isolate, and 98% with the Lebanon isolate. The results proved that the unusual disorder found in greenhouse tomatoes in Sicily can be associated with infections by PepMV and ToCV, reported for the first time in a mixed infection.

  5. Turbulent Region Near Jupiter's Great Red Spot

    Science.gov (United States)

    1997-01-01

    True and false color mosaics of the turbulent region west of Jupiter's Great Red Spot. The Great Red Spot is on the planetary limb on the right hand side of each mosaic. The region west (left) of the Great Red Spot is characterized by large, turbulent structures that rapidly change in appearance. The turbulence results from the collision of a westward jet that is deflected northward by the Great Red Spot into a higher latitude eastward jet. The large eddies nearest to the Great Red Spot are bright, suggesting that convection and cloud formation are active there.The top mosaic combines the violet (410 nanometers) and near infrared continuum (756 nanometers) filter images to create a mosaic similar to how Jupiter would appear to human eyes. Differences in coloration are due to the composition and abundance of trace chemicals in Jupiter's atmosphere. The lower mosaic uses the Galileo imaging camera's three near-infrared (invisible) wavelengths (756 nanometers, 727 nanometers, and 889 nanometers displayed in red, green, and blue) to show variations in cloud height and thickness. Light blue clouds are high and thin, reddish clouds are deep, and white clouds are high and thick. Purple most likely represents a high haze overlying a clear deep atmosphere. Galileo is the first spacecraft to distinguish cloud layers on Jupiter.The mosaic is centered at 16.5 degrees south planetocentric latitude and 85 degrees west longitude. The north-south dimension of the Great Red Spot is approximately 11,000 kilometers. The smallest resolved features are tens of kilometers in size. North is at the top of the picture. The images used were taken on June 26, 1997 at a range of 1.2 million kilometers (1.05 million miles) by the Solid State Imaging (SSI) system on NASA's Galileo spacecraft.The Jet Propulsion Laboratory, Pasadena, CA manages the Galileo mission for NASA's Office of Space Science, Washington, DC. JPL is an operating division of California Institute of Technology (Caltech

  6. Plasma spot welding of ferritic stainless steels

    Directory of Open Access Journals (Sweden)

    Lešnjak, A.

    2002-06-01

    Full Text Available Plasma spot welding of ferritic stainless steels is studied. The study was focused on welding parameters, plasma and shielding gases and the optimum welding equipment. Plasma-spot welded overlap joints on a 0.8 mm thick ferritic stainless steel sheet were subjected to a visual examination and mechanical testing in terms of tension-shear strength. Several macro specimens were prepared. Plasma spot welding is suitable to use the same gas as shielding gas and as plasma gas, i.e., a 98 % Ar/2 % H 2 gas mixture. Tension-shear strength of plasma-spot welded joints was compared to that of resistance-spot welded joints. It was found that the resistance welded joints withstand a somewhat stronger load than the plasma welded joints due to a larger weld spot diameter of the former. Strength of both types of welded joints is approximately the same.

    El artículo describe el proceso de soldeo de aceros inoxidables ferríticos por puntos con plasma. La investigación se centró en el establecimiento de los parámetros óptimos de la soldadura, la definición del gas de plasma y de protección más adecuado, así como del equipo óptimo para la realización de la soldadura. Las uniones de láminas de aceros inoxidables ferríticos de 0,8 mm de espesor, soldadas a solape por puntos con plasma, se inspeccionaron visualmente y se ensayaron mecánicamente mediante el ensayo de cizalladura por tracción. Se realizaron macro pulidos. Los resultados de la investigación demostraron que la solución más adecuada para el soldeo por puntos con plasma es elegir el mismo gas de plasma que de protección. Es decir, una mezcla de 98 % de argón y 2 % de hidrógeno. La resistencia a la cizalladura por tracción de las uniones soldadas por puntos con plasma fue comparada con la resistencia de las uniones soldadas por resistencia por puntos. Se llegó a la conclusión de que las uniones soldadas por resistencia soportan una carga algo mayor que la uniones

  7. Role Bending: Complex Relationships Between Viruses, Hosts, and Vectors Related to Citrus Leprosis, an Emerging Disease.

    Science.gov (United States)

    Roy, Avijit; Hartung, John S; Schneider, William L; Shao, Jonathan; Leon, Guillermo; Melzer, Michael J; Beard, Jennifer J; Otero-Colina, Gabriel; Bauchan, Gary R; Ochoa, Ronald; Brlansky, Ronald H

    2015-07-01

    Citrus leprosis complex is an emerging disease in the Americas, associated with two unrelated taxa of viruses distributed in South, Central, and North America. The cytoplasmic viruses are Citrus leprosis virus C (CiLV-C), Citrus leprosis virus C2 (CiLV-C2), and Hibiscus green spot virus 2, and the nuclear viruses are Citrus leprosis virus N (CiLV-N) and Citrus necrotic spot virus. These viruses cause local lesion infections in all known hosts, with no natural systemic host identified to date. All leprosis viruses were believed to be transmitted by one species of mite, Brevipalpus phoenicis. However, mites collected from CiLV-C and CiLV-N infected citrus groves in Mexico were identified as B. yothersi and B. californicus sensu lato, respectively, and only B. yothersi was detected from CiLV-C2 and CiLV-N mixed infections in the Orinoco regions of Colombia. Phylogenetic analysis of the helicase, RNA-dependent RNA polymerase 2 domains and p24 gene amino acid sequences of cytoplasmic leprosis viruses showed a close relationship with recently deposited mosquito-borne negevirus sequences. Here, we present evidence that both cytoplasmic and nuclear viruses seem to replicate in viruliferous Brevipalpus species. The possible replication in the mite vector and the close relationship with mosquito borne negeviruses are consistent with the concept that members of the genus Cilevirus and Higrevirus originated in mites and citrus may play the role of mite virus vector.

  8. Spot table - RPD | LSDB Archive [Life Science Database Archive metadata

    Lifescience Database Archive (English)

    Full Text Available switchLanguage; BLAST Search Image Search Home About Archive Update History Data ...d_spot.zip File URL: ftp://ftp.biosciencedbc.jp/archive/rpd/LATEST/rpd_spot.zip F... cDNA. (multiple entries) About This Database Database Description Download License Update History of This Database Site Policy | Contact Us Spot table - RPD | LSDB Archive ...

  9. 7 CFR 28.426 - Strict Good Ordinary Spotted Color.

    Science.gov (United States)

    2010-01-01

    ... 7 Agriculture 2 2010-01-01 2010-01-01 false Strict Good Ordinary Spotted Color. 28.426 Section 28.426 Agriculture Regulations of the Department of Agriculture AGRICULTURAL MARKETING SERVICE (Standards... Spotted Color. Strict Good Ordinary Spotted Color is color which is within the range represented by a set...

  10. 7 CFR 28.421 - Good Middling Spotted Color.

    Science.gov (United States)

    2010-01-01

    ... 7 Agriculture 2 2010-01-01 2010-01-01 false Good Middling Spotted Color. 28.421 Section 28.421 Agriculture Regulations of the Department of Agriculture AGRICULTURAL MARKETING SERVICE (Standards... Color. Good Middling Spotted Color is color which is better than Strict Middling Spotted Color. ...

  11. 7 CFR 28.424 - Strict Low Middling Spotted Color.

    Science.gov (United States)

    2010-01-01

    ... 7 Agriculture 2 2010-01-01 2010-01-01 false Strict Low Middling Spotted Color. 28.424 Section 28.424 Agriculture Regulations of the Department of Agriculture AGRICULTURAL MARKETING SERVICE (Standards... Spotted Color. Strict Low Middling Spotted Color is color which is within the range represented by a set...

  12. OBTENÇÃO DE ISOLADO PURO DE Anaplasma marginale EM BEZERROS NEONATOS PRIVADOS DE COLOSTRO Anaplasma marginale PURE ISOLATE OBTAINED IN COLOSTRUM-DEPRIVED NEWBORN CALVES

    Directory of Open Access Journals (Sweden)

    Francisco de Carvalho Dias Filho

    2008-07-01

    Full Text Available

    O trabalho foi realizado com o objetivo principal de obter isolados autóctones puros de Anaplasma marginale a partir de amostra de sangue de bovino criado em uma área endêmica para a tristeza parasitária bovina no município de Goiânia. Obteve-se o inóculo para o isolamento de A. marginale de animal doador, portador de infecções naturais mistas, após receber tratamento seletivo com dose babesicida esterelizante de dipropionato de imidocarb. Dois dias após esse tratamento, colheu-se dele uma amostra de sangue de 20 mL do animal doador para inoculação em bezerro neonato privado de colostro e livre de infecções por hemoparasitos. Após trinta dias da inoculação, o receptor apresentou febre, apatia e parasitemia patente por A. marginale. Amostras de sangue foram então colhidas e preparadas na forma de estabilizados para serem criopreservadas e, assim, comporem banco de isolados de hemoparasitos autóctones. Realizou-se a comprovação da pureza do isolado, concomitantemente, pela demonstração de soroconversão específica, pela reação de PCR e, ainda, pela subinoculação da amostra criopreservada em bezerro suscetível. Constatou-se ainda que o uso de bezerros neonatos privados de colostro, como animais suscetíveis, pode ser considerado como modelo prático, eficaz e relativamente barato para o isolamento de hemoparasitos em regiões endêmicas.

     

    PALAVRAS-CHAVES: Anaplasma marginale, anaplasmose bovina, bezerros, isolados.

    This study was conducted with the objective of obtaining pure autochthonous isolates of Anaplasma marginale from blood samples of bovines that have been raised in the tick-borne disease endemic area of the municipality of Goiânia. The inoculum of A. marginale was prepared from a donor animal, known to be natural carrier of hemoparasite mixed infections, after treatment with

  13. Estudo imunobiológico de estoques de Trypanosoma cruzi isolados de pacientes na fase aguda da doença de Chagas

    Directory of Open Access Journals (Sweden)

    T.L. Kipnis

    1983-12-01

    Full Text Available Estoques de tripanossomas isolados de pacientes na fase aguda da doença de Chagas foram injetados em grupos de camundongos albinos não isogênicos nas doses de 10³, 10(4 e 10(5 parasitas/camundongos. O curso da infecção foi seguido por três meses. A pctrasitemia foi em geral baixa, com picos recorrentes, na maioria das vezes os animais evoluiam para cronicidade. Somente um estoque induziu alto índice de mortalidade. Os parasitas e as lesões apesar de detectadas no pico da parasitemia e restritos ao coração estavam ausentes aos três meses. Nesta época os perfis de Igs apresentaram diferenças marcantes. Grupos de animais que foram inoculados com estes estoques foram desafiados com doses letais da cepa Y ou CL. Em alguns casos obteve-se uma parasitemia, mas patente.

  14. Caracterização fenogenotípica da resistência antimicrobiana em Staphylococcus spp. isolados de mastite bovina

    Directory of Open Access Journals (Sweden)

    Elaine C.L. Mendonça

    2012-09-01

    Full Text Available A utilização de antibióticos no controle das infecções intramamárias e na eliminação de prováveis fontes de infecção nas fazendas leiteiras se constitui em importante medida de controle. No entanto, o uso inadequado de antibióticos no tratamento da doença pode selecionar cepas resistentes e comprometer a eficiência do tratamento. Bactérias do gênero Staphylococcus spp. estão entre os principais agentes etiológicos da mastite bovina e são freqüentemente resistentes aos antimicrobianos, em especial aos beta-lactâmicos, principalmente por dois mecanismos distintos: a produção da enzima extracelular beta-lactamase, codificada pelo gene blaZ, e a produção de PBP2a ou PBP2´, uma proteína ligante de penicilina de baixa afinidade, codificada pelo gene mecA. A expressão do gene mecA é constitutiva ou induzida por antibióticos betalactâmicos, como a oxacilina e cefoxitina. O gene mecA está inserido no cromossomo através de um elemento genético móvel, denominado cassete estafilocócico cromossômico mec (SCCmec. O presente estudo avaliou o perfil fenogenotípico de resistência aos beta-lactâmicos em 250 isolados de Staphylococcus spp., utilizando os marcadores oxacilina e cefoxitina, de modo a produzir dados que possam contribuir para o conhecimento da resistência antimicrobiana em algumas propriedades leiteiras das regiões Sul-Fluminense e Metropolitana do Estado do Rio de Janeiro com o objetivo de subsidiar a implementação de medidas de controle dessa enfermidade. A avaliação da resistência foi feita a partir de 8 diferentes testes fenotípicos, sendo obtidos 54 perfis. Os testes de difusão em disco simples e ágar screen com oxacilina foram utilizados como "padrão ouro" para os cálculos dos valores de sensibilidade, especificidade e predição por serem preconizados pelo CLSI veterinário. O teste de difusão em disco simples com cefoxitina foi o de melhor desempenho na predição da resistência a

  15. Jupiter Great Red Spot and White Ovals

    Science.gov (United States)

    1979-01-01

    This photo of Jupiter was taken by Voyager 1 on March 1, 1979. The spacecraft was 3 million miles (5 million kilometers) from Jupiter at the time. The photo shows Jupiter's Great Red Spot (upper right) and the turbulent region immediately to the west. At the middle right of the frame is one of several white ovals seen on Jupiter from Earth. The structure in every feature here is far better than has ever been seen from any telescopic observations. The Red Spot and the white oval both reveal intricate and involved structure. The smallest details that can be seen in this photo are about 55 miles (95 kilometers) across. JPL manages and controls the Voyager project for NASA's Office of Space Science.

  16. Jupiter's Great Red Spot and White Ovals

    Science.gov (United States)

    1979-01-01

    This photo of Jupiter was taken by Voyager 1 on the evening of March 1, 1979, from a distance of 2.7 million miles (4.3 million kilometers). The photo shows Jupiter's Great Red Spot (top) and one of the white ovals than can be seen in Jupiter's atmosphere from Earth. The white ovals were seen to form in 1939, and 1940, and have remained more or less constant ever since. None of the structure and detail evident in these features have ever been seen from Earth. The Great Red Spot is three times as large as Earth. Also evident in the picture is a great deal of atmospheric detail that will require further study for interpretation. The smallest details that can be seen in this picture are about 45 miles (80 kilometers across. JPL manages and controls the Voyager project for NASA's Office of Space Science.

  17. spots de campaña

    Directory of Open Access Journals (Sweden)

    Julio Juárez Gámiz

    2007-01-01

    publicidad política en televisión. Ilustrado por los spots televisivos, el debate se ha centrado en el alto costo que representa para los contribuyentes, a través de los partidos políticos, la producción y transmisión de estos mensajes. El presente trabajo consiste en un análisis de contenido de una muestra de spots transmitidos por las tres principales fuerzas electorales en México durante la campaña presidencial de 2006. El objetivo es identificar, de manera sistemática, características particulares en el formato y contenido de estos mensajes a la luz de su función persuasiva e informativa.

  18. Experimental and simulated strength of spot welds

    DEFF Research Database (Denmark)

    Nielsen, Chris Valentin; Bennedbæk, Rune A.K.; Larsen, Morten B.

    2014-01-01

    Weld strength testing of single spots in DP600 steel is presented for the three typical testing procedures, i.e. tensile-shear, cross-tension and peel testing. Spot welds are performed at two sets of welding parameters and strength testing under these conditions is presented by load......-elongation curves revealing the maximum load and the elongation at break. Welding and strength testing is simulated by SORPAS® 3D, which allows the two processes to be prepared in a combined simulation, such that the simulated welding properties are naturally applied to the simulation of strength testing. Besides...... the size and shape of the weld nugget, these properties include the new strength of the material in the weld and the heat affected zone based on the predicted hardness resulting from microstructural phase changes simulated during cooling of the weld before strength testing. Comparisons between overall...

  19. Sweet spot supersymmetry and composite messengers

    Science.gov (United States)

    Ibe, Masahiro; Kitano, Ryuichiro

    2008-05-01

    Sweet spot supersymmetry is a phenomenological effective Lagrangian of weak scale supersymmetry with a certain set of natural assumptions. This framework is designed to avoid problems in low-energy phenomenology and cosmology of supersymmetric models. We discuss a class of dynamical models of supersymmetry breaking and its mediation, whose low-energy effective description falls into this framework. Hadron fields in the dynamical models play a role of the messengers of the supersymmetry breaking. As is always true in the models of the sweet spot supersymmetry, the messenger scale is predicted to be 105 GeV ≲Mmess ≲1010 GeV. Various values of the effective number of messenger fields Nmess are possible depending on the choice of the gauge group.

  20. Forecasting European thermal coal spot prices

    Directory of Open Access Journals (Sweden)

    Alicja Krzemień

    2015-01-01

    Finally, in order to analyse the time series model performance a Generalized Regression Neural Network (GRNN was used and its performance compared against the whole AR(2 process. Empirical results obtained confirmed that there is no statistically significant difference between both methods. The GRNN analysis also allowed pointing out the main drivers that move the European Thermal Coal Spot prices: crude oil, USD/CNY change and supply side drivers.