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Sample records for white spot virus

  1. Immune defence White Spot Syndrome Virus infected shrimp, Penaeus monodon

    OpenAIRE

    Arts, J. A. J.

    2006-01-01

    White spot syndrome virus (WSSV) is the most important viral pathogen of cultured penaeid shrimp worldwide. Since the initial discovery of the virus inTaiwanin 1992, it has spread to shrimp farming regions in Southeast Asia, theAmericas, Europe and theMiddle Eastcausing major economic losses. The virus has a wide host range among crustaceans and induces distinctive clinical signs (white spots)on the inner surface of the exoskeletonof penaeid shrimps.Limited data is available about the immune ...

  2. THE WHITE SPOT SYNDROME VIRUS (WSSV) LOAD IN Dendronereis spp.

    OpenAIRE

    Desrina; Sarjito; Alfabetian Harjuno Condro Haditomo; Diana Chilmawati

    2012-01-01

    The white spot syndrome virus (WSSV), the causative agent of White Spot Syndrome Disease (WSSD), is a major shrimp pathogen in Indonesia. Dendronereis spp. is a ubiquitous Polychaetes and natural food of shrimp raised in brackishwater pond in Indonesia. The objective of this research is to determine the occurrence of WSSV and the viral load in Dendronereis spp. obtained from the shrimp pond. Dendronereis spp. was obtained with PVC (10 cm in diameter) from a traditional shrimp pond in Semarang...

  3. White spot syndrome virus: Genotypes, Epidemiology and Evolutionary Studies

    OpenAIRE

    Shekar, Malathi; Pradeep, Balakrishnan; Karunasagar, Indrani

    2012-01-01

    White spot syndrome virus (WSSV) is a pathogen that has emerged globally affecting shrimp populations. Comparison of WSSV genome have shown the virus to share a high genetic similarity except for a few variable genomic loci that has been employed as markers in molecular epidemiology studies for determining the origin, evolution and spread in different geographical regions. Molecular genotyping of WSSV are based on genomic deletions associated with ORF23/24 and ORF14/15 variable regions and th...

  4. Genetic variation among isolates of White Spot Syndrome Virus

    OpenAIRE

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

    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 (RFLP) pattern. We have mapped the genomic differences between three completely sequenced WSSV isolates, originating from Thailand (WSSV-TH), China (WSSV-CN) and Taiwan (WSSV-TW). Alignment of the g...

  5. White spot syndrome virus inactivation study by using gamma irradiation

    Science.gov (United States)

    Heidareh, Marzieh; Sedeh, Farahnaz Motamedi; Soltani, Mehdi; Rajabifar, Saeed; Afsharnasab, Mohammad; Dashtiannasab, Aghil

    2014-09-01

    The present study was conducted to investigate the effect of gamma irradiation on white spot syndrome virus (WSSV). White spot syndrome virus is a pathogen of major economic importance in cultured penaeid shrimp industries. White spot disease can cause mortalities reaching 100% within 3-10 days of gross signs appearing. During the period of culture, immunostimulant agents and vaccines may provide potential methods to protect shrimps from opportunistic and pathogenic microrganisms. In this study, firstly, WSSV was isolated from infected shrimp and then multiplied in crayfish. WSSV was purified from the infected crayfish haemolymph by sucrose gradient and confirmed by transmission electron microscopy. In vivo virus titration was performed in shrimp, Penaeus semisulcatus. The LD50 of live virus stock was calculated 10 5.4/mL. Shrimp post-larvae (1-2 g) were treated with gamma-irradiated (different doses) WSSV (100 to 10-4 dilutions) for a period of 10 days. The dose/survival curve for irradiated and un-irradiated WSSV was drawn; the optimum dose range for inactivation of WSSV and unaltered antigenicity was obtained 14-15 kGy. This preliminary information suggests that shrimp appear to benefit from treatment with gammairradiated WSSV especially at 14-15 KGy.

  6. The white spot syndrome virus DNA genome sequence

    OpenAIRE

    Hulten, M.C.W., van; Witteveldt, J.; Peters, S; Kloosterboer, N.; Tarchini, R.; M. Fiers; Sandbrink, H.; Klein Lankhorst, R.; J. M. Vlak

    2001-01-01

    White spot syndrome virus (WSSV) is at present a major scourge to worldwide shrimp cultivation. We have determined the entire sequence of the double-stranded, circular DNA genome of WSSV, which contains 292,967 nucleotides encompassing 184 major open reading frames (ORFs). Only 6 f the WSSV ORFs have putative homologues in databases, mainly representing genes encoding enzymes for nucleotide metabolism, DNA replication, and protein modification. The remaining ORFs are mostly unassigned, except...

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

    OpenAIRE

    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 towards the presumed less susceptibleLitopenaeusvannamei , the use of specific pathogen free shrimp and the development of more advanced shrimp culturing techniques, WSSV continues to scourge shrimp farms. Therefore there is an urgent need for effective intervention strategies. Vac...

  8. Genomics and transcriptomics of White spot syndrome virus

    OpenAIRE

    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 genome of WSSV ranges between 293 and 307 kb in size depending on the WSSV isolate. For a sequenced isolate originating fromThailand(WSSV-TH) 184 putative open reading frames (ORFs) were identified ...

  9. THE WHITE SPOT SYNDROME VIRUS (WSSV LOAD IN Dendronereis spp.

    Directory of Open Access Journals (Sweden)

    Desrina

    2012-06-01

    Full Text Available The white spot syndrome virus (WSSV, the causative agent of White Spot Syndrome Disease (WSSD, is a major shrimp pathogen in Indonesia. Dendronereis spp. is a ubiquitous Polychaetes and natural food of shrimp raised in brackishwater pond in Indonesia. The objective of this research is to determine the occurrence of WSSV and the viral load in Dendronereis spp. obtained from the shrimp pond. Dendronereis spp. was obtained with PVC (10 cm in diameter from a traditional shrimp pond in Semarang vicinity. As acomparison, healthy looking Penaeus monodon was also obtained from the same pond. The occurrence of WSSV in Dendronereis spp. was determined with 1-step and nested PCR using primer for WSSV major envelope protein, VP 28. The viral load was counted with 1-step Real Time PCR. The WSSV was detected in Dendronereis spp. with 1-step and nested PCR. The point prevalence of WSSV infection in Dendronereis spp. is 90 %. The viral load ranged from 0 to 1.9 x 104 copy of DNA/µg total DNA. The viral load in Dendronereis is comparable with that of naturally infected and at carrier state P.monodon from the same pond. This is the first report of WSSV load in naturally infected Dendronereis spp.

  10. Suppression of shrimp melanization during white spot syndrome virus infection.

    Science.gov (United States)

    Sutthangkul, Jantiwan; Amparyup, Piti; Charoensapsri, Walaiporn; Senapin, Saengchan; Phiwsaiya, Kornsunee; Tassanakajon, Anchalee

    2015-03-01

    The melanization cascade, activated by the prophenoloxidase (proPO) system, plays a key role in the production of cytotoxic intermediates, as well as melanin products for microbial sequestration in invertebrates. Here, we show that the proPO system is an important component of the Penaeus monodon shrimp immune defense toward a major viral pathogen, white spot syndrome virus (WSSV). Gene silencing of PmproPO(s) resulted in increased cumulative shrimp mortality after WSSV infection, whereas incubation of WSSV with an in vitro melanization reaction prior to injection into shrimp significantly increased the shrimp survival rate. The hemolymph phenoloxidase (PO) activity of WSSV-infected shrimp was extremely reduced at days 2 and 3 post-injection compared with uninfected shrimp but was fully restored after the addition of exogenous trypsin, suggesting that WSSV probably inhibits the activity of some proteinases in the proPO cascade. Using yeast two-hybrid screening and co-immunoprecipitation assays, the viral protein WSSV453 was found to interact with the proPO-activating enzyme 2 (PmPPAE2) of P. monodon. Gene silencing of WSSV453 showed a significant increase of PO activity in WSSV-infected shrimp, whereas co-silencing of WSSV453 and PmPPAE2 did not, suggesting that silencing of WSSV453 partially restored the PO activity via PmPPAE2 in WSSV-infected shrimp. Moreover, the activation of PO activity in shrimp plasma by PmPPAE2 was significantly decreased by preincubation with recombinant WSSV453. These results suggest that the inhibition of the shrimp proPO system by WSSV partly occurs via the PmPPAE2-inhibiting activity of WSSV453. PMID:25572398

  11. Experiments on the effect of temperature on white spot syndrome virus infection in Litopenaeus vannamei shrimp

    OpenAIRE

    Lima, João José Pereira Dantas da Rocha

    2006-01-01

    White Spot Disease (WSD) is an aggressive and devastating viral disease caused by the White Spot Syndrome Virus (WSSV). This highly pathogenic and widespread disease, present throughout Asia and the Americas, can cause up to 100% mortality within 3-7 days after infection. It is annually responsible for huge ecological and economical losses in the main producing countries and forms as such one of the greatest threats for the further sustainable development of shrimp aquaculture. Previous re...

  12. VP292 of White spot syndrome virus Interacts with VP26

    OpenAIRE

    LI, Qian; Liu, Qing-Hui; Huang, Jie

    2012-01-01

    Interactions between virus structural proteins are suggested to be crucial for virus assembly. Many steps in the process of white spot syndrome virus (WSSV) assembly and maturation remain unclear. In this paper, we discovered a new interaction of WSSV VP292. Temporal-transcription analysis showed that VP292 is expressed in the late stage of WSSV infection. Western blot and matrix-assisted laser desorption ionization MS assays showed that VP292 interacts with VP26, a major envelope protein. Fa...

  13. Fitness and virulence of an ancestral White Spot Syndrome Virus isolate from shrimp

    OpenAIRE

    Marks, H.; Duijse, J.J.A.; Zuidema, D.; Hulten, M.C.W., van; J. M. Vlak

    2005-01-01

    White Spot Syndrome Virus, the type species of the virus family Nimaviridae, is a large dsDNA virus infecting shrimp and other crustaceans. Genomic analysis of three completely sequenced WSSV isolates identified two major polymorphic loci, ¿variable region ORF14/15¿ and ¿variable region ORF23/24¿. Here, we characterize a WSSV isolate originating from shrimp collected in Thailand in 1996 (TH-96-II). This isolate contains the largest WSSV genome (312 kb) identified so far, mainly because of its...

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

    OpenAIRE

    Sandbrink Hans; Ren Xin-Ying; Marks Hendrik; van Hulten Mariëlle CW; Vlak Just M

    2006-01-01

    Abstract 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 cascaded fashion. To search in silico for conserved promoter motifs (i) the abundance of all 4 through 8 nucleotide motifs in the upstream sequences of WSSV genes relative to the complete ge...

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

    OpenAIRE

    Hulten, M.C.W., van

    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 further into Central- and South-America. In cultured shrimp WSSV infection can reach a cumulative mortality of up to 100% within 3 to 10 days.One of the clinical signs of WSSV is the appearance of white spots in the exoskeleton of ...

  16. Biology, Host Range, Pathogenesis and Diagnosis of White spot syndrome virus

    OpenAIRE

    Pradeep, Balakrishnan; Rai, Praveen; Mohan, Seethappa A.; Shekhar, Mudagandur S.; Karunasagar, Indrani

    2012-01-01

    White spot syndrome virus (WSSV) is the most serious viral pathogen of cultured shrimp. It is a highly virulent virus that can spread quickly and can cause up to 100 % mortality in 3–10 days. WSSV is a large enveloped double stranded DNA virus belonging to genus Whispovirus of the virus family Nimaviridae. It has a wide host range among crustaceans and mainly affects commercially cultivated marine shrimp species. The virus infects all age groups causing large scale mortalities and the foci of...

  17. Susceptibility of testicular cell cultures of crab, Scylla serrata (Forskal) to white spot syndrome virus

    OpenAIRE

    Shashikumar, Anumol; Desai, P. V.

    2012-01-01

    Testicular cell culture of crab, Scylla serrata (Forskal) was used to study the effects of White spot syndrome virus (WSSV). We are showing the susceptibility of cell culture of crabs to WSSV. The proliferating cell culture of testes were maintained for more than 4 months in a medium prepared from L15 and crab saline supplemented with epidermal growth factor. The cell cultures inoculated with different concentrations of virus showed distinct cytopathic effects such as change in cell appearanc...

  18. Construction and Application of a Protein Interaction Map for White Spot Syndrome Virus (WSSV)*

    OpenAIRE

    Sangsuriya, Pakkakul; Huang, Jiun-Yan; Chu, Yu-Fei; Phiwsaiya, Kornsunee; Leekitcharoenphon, Pimlapas; Meemetta, Watcharachai; Senapin, Saengchan; Huang, Wei-Pang; Withyachumnarnkul, Boonsirm; Flegel, Timothy W; Lo, Chu-Fang

    2013-01-01

    White spot syndrome virus (WSSV) is currently the most serious global threat for cultured shrimp production. Although its large, double-stranded DNA genome has been completely characterized, most putative protein functions remain obscure. To provide more informative knowledge about this virus, a proteomic-scale network of WSSV-WSSV protein interactions was carried out using a comprehensive yeast two-hybrid analysis. An array of yeast transformants containing each WSSV open reading frame fused...

  19. Gene-expression profiling of White spot syndrome virus in vivo

    OpenAIRE

    Marks, H.; Vorst, O.F.J.; Houwelingen, A.M.M.L., van; Hulten, M.C.W., van; J. M. Vlak

    2005-01-01

    White spot syndrome virus, type species of the genus Whispovirus in the family Nimaviridae, is a large, double-stranded DNA (dsDNA) virus that infects crustaceans. The genome of the completely sequenced isolate WSSV-TH encodes 184 putative open reading frames (ORFs), the functions of which are largely unknown. To study the transcription of these ORFs, a DNA microarray was constructed, containing probes corresponding to nearly all putative WSSV-TH ORFs. Transcripts of 79 % of these ORFs could ...

  20. Characterization and Interactome Study of White Spot Syndrome Virus Envelope Protein VP11

    OpenAIRE

    Liu, Wang-Jing; Shiung, Hui-Jui; Lo, Chu-Fang; Leu, Jiann-Horng; Lai, Ying-Jang; Lee, Tai-Lin; Huang, Wei-Tung; Kou, Guang-Hsiung; Chang, Yun-Shiang

    2014-01-01

    White spot syndrome virus (WSSV) is a large enveloped virus. The WSSV viral particle consists of three structural layers that surround its core DNA: an outer envelope, a tegument and a nucleocapsid. Here we characterize the WSSV structural protein VP11 (WSSV394, GenBank accession number AF440570), and use an interactome approach to analyze the possible associations between this protein and an array of other WSSV and host proteins. Temporal transcription analysis showed that vp11 is an early g...

  1. Detection of white spot syndrome virus (WSSV) of Penaeus chinensis by in situ hybridization

    Science.gov (United States)

    Zhan, Wen-Bin; Wang, Yuan-Hong; Zhang, Zhi-Dong; Hideo, Fukuda

    2000-09-01

    White Spot Syndrome Virus (WSSV) was purified from hemolymph of infected shrimp. After nucleic acid extraction from the purified virus particles, EcoR I-digested fragments of the WSSV genome were cloned; three of these fragments were used as non-radioactive probes labeled with DIG-11-dUTP. The probes hybridized in situ, with sections located in the nuclei of all WSSV-infected tissues. The virus was detected in the gill, stomach, epidermis, and connective tissue and so on, but not detected in healthy shrimp tissues and epithelial cells of hepatopancreatic tubules of diseased shrimp.

  2. White spot syndrome virus envelope protein VP28 is involved in the systemic infection of shrimp

    OpenAIRE

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

    2001-01-01

    White spot syndrome virus (WSSV) is a large DNA virus infecting shrimp and other crustaceans. The virus particles contain at least five major virion proteins, of which three (VP26, VP24, and VP15) are present in the rod-shaped nucleocapsid and two (VP28 and VP19) reside in the envelope. The mode of entry and systemic infection of WSSV in the black tiger shrimp, Penaeus monodon, and the role of these proteins in these processes are not known. A specific polyclonal antibody was generated agains...

  3. Evaluation of white spot syndrome virus variable DNA loci as molecular markers of virus spread at intermediate spatiotemporal scales

    OpenAIRE

    Bui Thi Minh Dieu; Marks, H.; Zwart, M.P.; J. M. Vlak

    2010-01-01

    Variable genomic loci have been employed in a number of molecular epidemiology studies of white spot syndrome virus (WSSV), but it is unknown which loci are suitable molecular markers for determining WSSV spread on different spatiotemporal scales. Although previous work suggests that multiple introductions of WSSV occurred in central Vietnam, it is largely uncertain how WSSV was introduced and subsequently spread. Here, we evaluate five variable WSSV DNA loci as markers of virus spread on an ...

  4. Membrane Binding Activity with Virus in White Spot Syndrome Virus-Infected Fenneropenaeus chinensis

    Directory of Open Access Journals (Sweden)

    Liu Qing-hui

    2009-01-01

    Full Text Available Problem statement: White Spot Syndrome Virus (WSSV is one of the most disastrous pathogens in shrimp culture, having caused high mortality in many cultured shrimp species. It is well known that the most important step of viral infection is cell attachment. Approach:Therefore determining the binding activity of shrimp cells to WSSV is an important to evaluate anti-infection ability and understanding the procedure of a viral infection. So this study aims to analyze binding activity of cell membrane with WSSV in shrimp of Fenneropenaeus chinensis (F. chienesis. WSSV was used to stimulate F. chinensis and binding activity in gills, muscle and hepatopancreas with virus within the procession of WSSV was measured. Results:The results showed that binding activity in gills with WSSV within infection stage (0-96 h had no significant change. The highest binding activity was attained after infection 24-48 h in muscle. The binding activity in hepatopancreas with WSSV was lower during infection stage (0-96 h. Conclusion/Recommendation: The higher binding activity of WSSV to gills and muscle indicated that gills and muscle are the importance target tissue for WSSV infection. This suggested that receptor of WSSV existed in gills and muscle.

  5. Transcriptional analysis of the ribonucleotide reductase genes in shrimp white spot syndrome virus

    OpenAIRE

    Tsai, M.F.; Lo, C F; Hulten, M.C.W., van; Tzeng, H.F.; Chou, C.M.; Huang, C. J.; Wang, C.S.

    2000-01-01

    The causative agent of white spot syndrome (WSS) is a large double-stranded DNA virus, WSSV, which is probably a representative of a new genus, provisionally called Whispovirus. From previously constructed WSSV genomic libraries of a Taiwan WSSV isolate, clones with open reading frames (ORFs) that encode proteins with significant homology to the class I ribonucleotide reductase large (RR1) and small (RR2) subunits were identified. WSSV rr1 and rr2 potentially encode 848 and 413 amino acids, r...

  6. Genomic and Proteomic Analysis of Thirty-Nine Structural Proteins of Shrimp White Spot Syndrome Virus

    OpenAIRE

    Tsai, Jyh-Ming; Wang, Han-Ching; Leu, Jiann-Horng; Hsiao, He-Hsuan; Wang, Andrew H.-J.; Kou, Guang-Hsiung; Lo, Chu-Fang

    2004-01-01

    White spot syndrome virus (WSSV) virions were purified from the hemolymph of experimentally infected crayfish Procambarus clarkii, and their proteins were separated by 8 to 18% gradient sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) to give a protein profile. The visible bands were then excised from the gel, and following trypsin digestion of the reduced and alkylated WSSV proteins in the bands, the peptide sequence of each fragment was determined by liquid chromatograph...

  7. Proteomic Analysis of the Major Envelope and Nucleocapsid Proteins of White Spot Syndrome Virus?

    OpenAIRE

    Xie, Xixian; Xu, Limei; Feng YANG

    2006-01-01

    White spot syndrome virus (WSSV) virions were purified from the tissues of infected Procambarus clarkii (crayfish) isolates. Pure WSSV preparations were subjected to Triton X-100 treatment to separate into the envelope and nucleocapsid fractions, which were subsequently separated by 12% sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The major envelope and nucleocapsid proteins were identified by either matrix-assisted laser desorption ionization-time of flight mass spectrometry or...

  8. Molt stage and cuticle damage influence White Spot Syndrome Virus immersion infection in penaeid shrimp

    OpenAIRE

    Corteel, Mathias; Dantas-lima, Joa?o J.; Wille, Mathieu; Alday-sanz, Victoria; Pensaert, Maurice B.; Sorgeloos, Patrick; Nauwynck, Hans J.

    2009-01-01

    Abstract Transmission of white spot syndrome virus (WSSV) in shrimp has been reported to occur by feeding and immersion. In the present study, the impact of the molt process and artificial lesions in the cuticle on shrimp susceptibility to WSSV was examined using intramuscular and immersion routes. For the intramuscular route, Penaeus (Litopenaeus) vannamei shrimp (n=450) were injected with 10-2.3 up to 102.7 shrimp infectious dose 50% end point (SID50) of WSSV in early ...

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

    OpenAIRE

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

    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. The double-stranded viral DNA has an approximate size 290 kb. WSSV virions, isolated from infected shrimps, contained four major proteins: 28 kDa (VP28), 26 kDa (VP26), 24 kDa (VP24), and 19 kDa (...

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

    OpenAIRE

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

    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 associated with the nucleocapsid and the remaining two with the envelope. Forty-one N-terminal amino acids of VP24 were determined biochemically allowing the identification of its gene (vp24) in th...

  11. Hijacking of Host Calreticulin Is Required for the White Spot Syndrome Virus Replication Cycle

    OpenAIRE

    Watthanasurorot, Apiruck; Guo, Enen; Tharntada, Sirinit; Lo, Chu-Fang; Söderhäll, Kenneth; Söderhäll, Irene

    2014-01-01

    We have previously shown that multifunctional calreticulin (CRT), which resides in the endoplasmic reticulum (ER) and is involved in ER-associated protein processing, responds to infection with white spot syndrome virus (WSSV) by increasing mRNA and protein expression and by forming a complex with gC1qR and thereby delaying apoptosis. Here, we show that CRT can directly interact with WSSV structural proteins, including VP15 and VP28, during an early stage of virus infection. The binding of VP...

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

    OpenAIRE

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

    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 (VP28). The identified PK shows a high degree of homology to other viral and eukaryotic PK genes. Homology in the catalytic domains suggests that this PK is a serine/threonine protein kinase. All of the ...

  13. Enzyme E2 from Chinese White Shrimp Inhibits Replication of White Spot Syndrome Virus and Ubiquitinates Its RING Domain Proteins ? †

    OpenAIRE

    Chen, An-Jing; WANG, SHUAI; Zhao, Xiao-Fan; Yu, Xiao-Qiang; Wang, Jin-Xing

    2011-01-01

    Recent studies have shown that the ubiquitin (Ub) proteasome pathway (UPP) is closely related to immune defense. We have identified a ubiquitin-conjugating enzyme, E2, from the Chinese white shrimp, Fenneropenaeus chinensis (FcUbc). Injection of recombinant FcUbc protein (rFcUbc) reduced the mortality of shrimp infected with white spot syndrome virus (WSSV) and inhibited replication of WSSV. rFcUbc, but not a mutant FcUbc (mFcUbc), bound to WSSV RING domains (WRDs) from four potential E3 liga...

  14. Membrane Binding Activity with Virus in White Spot Syndrome Virus-Infected Fenneropenaeus chinensis

    OpenAIRE

    Liu Qing-hui; Huang Jie; Chen Ting; Yang Bing

    2009-01-01

    Problem statement: White Spot Syndrome Virus (WSSV) is one of the most disastrous pathogens in shrimp culture, having caused high mortality in many cultured shrimp species. It is well known that the most important step of viral infection is cell attachment. Approach:Therefore determining the binding activity of shrimp cells to WSSV is an important to evaluate anti-infection ability and understanding the procedure of a viral infection. So this study aims to ...

  15. Penaeus monodon Thioredoxin Restores the DNA Binding Activity of Oxidized White Spot Syndrome Virus IE1

    OpenAIRE

    Huang, Jiun-Yan; Liu, Wang-Jing; Wang, Han-Ching; Lee, Der-Yen; Leu, Jiann-Horng; Wang, Hao-Ching; Tsai, Mong-Hsun; Kang, Shih-Ting; Chen, I-Tung; Kou, Guang-Hsiung; Chang, Geen-Dong; Lo, Chu-Fang

    2012-01-01

    Aims: In this study we identified viral gene targets of the important redox regulator thioredoxin (Trx), and explored in depth how Trx interacts with the immediate early gene #1 (IE1) of the white spot syndrome virus (WSSV). Results: In a pull-down assay, we found that recombinant Trx bound to IE1 under oxidizing conditions, and a coimmunoprecipitation assay showed that Trx bound to WSSV IE1 when the transfected cells were subjected to oxidative stress. A pull-down assay with Trx mutants show...

  16. Four Major Envelope Proteins of White Spot Syndrome Virus Bind To Form a Complex? †

    OpenAIRE

    Qing ZHOU; Xu, Limei; Li, Hui; Qi, Yi-Peng; Feng YANG

    2009-01-01

    Early events in white spot syndrome virus (WSSV) morphogenesis, particularly the formation of viral membranes, are poorly understood. The major envelope proteins of WSSV are VP28, VP26, VP24, and VP19. Our previous results indicated that VP28 interacts with VP26 and VP24. In the present study, we used coimmunoprecipitation assays and pull-down assays to confirm that the four major proteins in the WSSV envelope can form a multiprotein complex. Yeast two-hybrid assays were also used to test for...

  17. Evolutionary Trajectory of White Spot Syndrome Virus (WSSV) Genome Shrinkage during Spread in Asia

    OpenAIRE

    Zwart, Mark P.; Dieu, Bui Thi Minh; Hemerik, Lia; Vlak, Just 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 occur at two loci in the WSSV genome, the ORF14/15 and ORF23/24 variable regions, and these have been used as molecular markers to study patterns of viral spread over space and time. We describe the ...

  18. White spot syndrome virus (WSSV) infection in shrimp (Litopenaeus vannamei) exposed to low and high salinity.

    Science.gov (United States)

    Ramos-Carreño, Santiago; Valencia-Yáñez, Ricardo; Correa-Sandoval, Francisco; Ruíz-García, Noé; Díaz-Herrera, Fernando; Giffard-Mena, Ivone

    2014-09-01

    White spot syndrome virus (WSSV) has a worldwide distribution and is considered one of the most pathogenic and devastating viruses to the shrimp industry. A few studies have explored the effect of WSSV on shrimp acclimated to low (5 practical salinity units [psu]) or high ([40 psu) salinity conditions. In this work, we analysed the physiological response of WSSV-infected Litopenaeus vannamei juveniles that were acclimated to different salinities (5, 15, 28, 34 and 54 psu). We evaluated the osmotic response and survival of the shrimp at different times after infection (0 to 48 hours), and we followed the expression levels of a viral gene (vp664) in shrimp haemolymph using real-time PCR. Our results indicate that the susceptibility of the shrimp to the virus increased at extreme salinities (5 and 54 psu), with higher survival rates at 15 and 28 psu, which were closer to the iso-osmotic point (24.7 psu, 727.5 mOsmol/kg). Acute exposure to the virus made the haemolymph less hyperosmotic at 5 and 15 psu and less hypo-osmotic at higher salinities ([28 psu). The capacity of white shrimp to osmoregulate, and thus survive, significantly decreased following WSSV infection. According to our results, extreme salinities (5 or 54 psu) are more harmful than seawater. PMID:24658782

  19. Prevalence and distribution of White Spot Syndrome Virus in cultured shrimp.

    Science.gov (United States)

    Hossain, A; Nandi, S P; Siddique, M A; Sanyal, S K; Sultana, M; Hossain, M A

    2015-02-01

    White Spot Syndrome Virus (WSSV) is a dsDNA virus causing White Spot Syndrome Disease (WSSD) in shrimp with almost 100% morality rate within 3-10 days. In Bangladesh, WSSD is one of the major impediments of shrimp farming. This study first investigated the prevalence and distribution of WSSV in cultured shrimps of the coastal regions in Bangladesh. A total of 60 shrimp samples, collected from the 25 shrimp farms of different coastal regions (Satkhira, Khulna, Bagerhat and Cox's Bazar), were analysed during 2013-2014 by conventional PCR using VP28 and VP664 gene-specific primers; 39 of 60 samples were found WSSV positive. SYBR green real-time PCR using 71-bp amplicon for VP664 gene correlated well with conventional PCR data. The prevalence rates of WSSV among the collected 60 samples were Satkhira 79%, Khulna 50%, Bagerhat 38% and Cox's Bazar 25%. Sequencing of WSSV-positive PCR amplicons of VP28 showed 99% similarity with WSSV NCBI Ref/Seq Sequences. Molecular analysis of the VP28 gene sequences of WSSV revealed that Bangladeshi strains phylogenetically affiliated to the strains belong to India. This work concluded that WSSV infections are widely distributed in the coastal regions cultured shrimp in Bangladesh. PMID:25402810

  20. Screening, isolation and optimization of anti–white spot syndrome virus drug derived from terrestrial plants

    Science.gov (United States)

    Ghosh, Upasana; Chakraborty, Somnath; Balasubramanian, Thangavel; Das, Punyabrata

    2014-01-01

    Objective To screen, isolate and optimize anti-white spot syndrome virus (WSSV) drug derived from various terrestrial plants and to evaluate the efficacy of the same in host–pathogen interaction model. Methods Thirty plants were subjected to Soxhlet extraction using water, ethanol, methanol and hexane as solvents. The 120 plant isolates thus obtained were screened for their in vivo anti–WSSV property in Litopenaeus vannamei. The best anti–WSSV plant isolate, TP22C was isolated and further analyzed. The drug was optimized at various concentrations. Viral and immune genes were analysed using reverse transcriptase PCR to confirm the potency of the drug. Results Seven plant isolates exhibited significant survivability in host. The drug TP22C thus formulated showed 86% survivability in host. The surviving shrimps were nested PCR negative at the end of the 15 d experimentation. The lowest concentration of TP22C required intramuscularly for virucidal property was 10 mg/mL. The oral dosage of 750 mg/kg body weight/day survived at the rate of 86%. Neither VP28 nor ie 1 was expressed in the test samples at 42nd hour and 84th hour post viral infection. Conclusions The drug TP22C derived from Momordica charantia is a potent anti-white spot syndrome virus drug. PMID:25183066

  1. White Spot Syndrome Virus Orf514 Encodes a Bona Fide DNA Polymerase

    Directory of Open Access Journals (Sweden)

    Rogerio R. Sotelo-Mundo

    2011-01-01

    Full Text Available White spot syndrome virus (WSSV is the causative agent of white spot syndrome, one of the most devastating diseases in shrimp aquaculture. The genome of WSSV includes a gene that encodes a putative family B DNA polymerase (ORF514, which is 16% identical in amino acid sequence to the Herpes virus 1 DNA polymerase. The aim of this work was to demonstrate the activity of the WSSV ORF514-encoded protein as a DNA polymerase and hence a putative antiviral target. A 3.5 kbp fragment encoding the conserved polymerase and exonuclease domains of ORF514 was overexpressed in bacteria. The recombinant protein showed polymerase activity but with very low level of processivity. Molecular modeling of the catalytic protein core encoded in ORF514 revealed a canonical polymerase fold. Amino acid sequence alignments of ORF514 indicate the presence of a putative PIP box, suggesting that the encoded putative DNA polymerase may use a host processivity factor for optimal activity. We postulate that WSSV ORF514 encodes a bona fide DNA polymerase that requires accessory proteins for activity and maybe target for drugs or compounds that inhibit viral DNA replication.

  2. 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

    2015-03-01

    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. PMID:25751859

  3. Rapid Detection of Shrimp White Spot Syndrome Virus by Real Time, Isothermal Recombinase Polymerase Amplification Assay

    Science.gov (United States)

    Xia, Xiaoming; Yu, Yongxin; Weidmann, Manfred; Pan, Yingjie; Yan, Shuling; Wang, Yongjie

    2014-01-01

    White spot syndrome virus (WSSV) causes large economic losses to the shrimp aquaculture industry, and thus far there are no efficient therapeutic treatments available against this lethal virus. In this study, we present the development of a novel real time isothermal recombinase polymerase amplification (RPA) assay for WSSV detection on a small ESEQuant Tube Scanner device. The RPA sensitivity, specificity and rapidity were evaluated by using a plasmid standard as well as viral and shrimp genomic DNAs. Compared with qPCR, the RPA assay revealed more satisfactory performance. It reached a detection limit up to 10 molecules in 95% of cases as determined by probit analysis of 8 independent experiments within 6.41±0.17 min at 39°C. Consequently, this rapid RPA method has great application potential for field use or point of care diagnostics. PMID:25121957

  4. DNA condensates organized by the capsid protein VP15 in White Spot Syndrome Virus

    International Nuclear Information System (INIS)

    The White Spot Syndrome Virus (WSSV) has a large circular double-stranded DNA genome of around 300 kb and it replicates in the nucleus of the host cells. The machinery of how the viral DNA is packaged has been remained unclear. VP15, a highly basic protein, is one of the major capsid proteins found in the virus. Previously, it was shown to be a DNA binding protein and was hypothesized to participate in the viral DNA packaging process. Using Atomic Force Microscopy imaging, we show that the viral DNA is associated with a (or more) capsid proteins. The organized viral DNA qualitatively resembles the conformations of VP15 induced DNA condensates in vitro. Furthermore, single-DNA manipulation experiments revealed that VP15 is able to condense single DNA against forces of a few pico Newtons. Our results suggest that VP15 may aid in the viral DNA packaging process by directly condensing DNA.

  5. The endemic copepod Calanus pacificus californicus as a potential vector of white spot syndrome virus.

    Science.gov (United States)

    Mendoza-Cano, Fernando; Sánchez-Paz, Arturo; Terán-Díaz, Berenice; Galván-Alvarez, Diego; Encinas-García, Trinidad; Enríquez-Espinoza, Tania; Hernández-López, Jorge

    2014-06-01

    The susceptibility of the endemic copepod Calanus pacificus californicus to white spot syndrome virus (WSSV) was established by the temporal analysis of WSSV VP28 transcripts by quantitative real-time PCR (qRT-PCR). The copepods were collected from a shrimp pond located in Bahia de Kino Sonora, Mexico, and challenged per os with WSSV by a virus-phytoplankton adhesion route. Samples were collected at 0, 24, 48 and 84 h postinoculation (hpi). The VP28 transcripts were not detected at early stages (0 and 24 hpi); however, some transcript accumulation was observed at 48 hpi and gradually increased until 84 hpi. Thus, these results clearly show that the copepod C. pacificus californicus is susceptible to WSSV infection and that it may be a potential vector for the dispersal of WSSV. However, further studies are still needed to correlate the epidemiological outbreaks of WSSV with the presence of copepods in shrimp ponds. PMID:24895865

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

    OpenAIRE

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

    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. 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...

  7. Comparative analysis of differentially expressed genes in normal and white spot syndrome virus infected Penaeus monodon

    Directory of Open Access Journals (Sweden)

    Juan Hsueh-Fen

    2007-05-01

    Full Text Available Abstract Background White spot syndrome (WSS is a viral disease that affects most of the commercially important shrimps and causes serious economic losses to the shrimp farming industry worldwide. However, little information is available in terms of the molecular mechanisms of the host-virus interaction. In this study, we used an expressed sequence tag (EST approach to observe global gene expression changes in white spot syndrome virus (WSSV-infected postlarvae of Penaeus monodon. Results Sequencing of the complementary DNA clones of two libraries constructed from normal and WSSV-infected postlarvae produced a total of 15,981 high-quality ESTs. Of these ESTs, 46% were successfully matched against annotated genes in National Center of Biotechnology Information (NCBI non-redundant (nr database and 44% were functionally classified using the Gene Ontology (GO scheme. Comparative EST analyses suggested that, in postlarval shrimp, WSSV infection strongly modulates the gene expression patterns in several organs or tissues, including the hepatopancreas, muscle, eyestalk and cuticle. Our data suggest that several basic cellular metabolic processes are likely to be affected, including oxidative phosphorylation, protein synthesis, the glycolytic pathway, and calcium ion balance. A group of immune-related chitin-binding protein genes is also likely to be strongly up regulated after WSSV infection. A database containing all the sequence data and analysis results is accessible at http://xbio.lifescience.ntu.edu.tw/pm/. Conclusion This study suggests that WSSV infection modulates expression of various kinds of genes. The predicted gene expression pattern changes not only reflect the possible responses of shrimp to the virus infection but also suggest how WSSV subverts cellular functions for virus multiplication. In addition, the ESTs reported in this study provide a rich source for identification of novel genes in shrimp.

  8. Hijacking of Host Calreticulin Is Required for the White Spot Syndrome Virus Replication Cycle

    Science.gov (United States)

    Watthanasurorot, Apiruck; Guo, Enen; Tharntada, Sirinit; Lo, Chu-Fang; Söderhäll, Kenneth

    2014-01-01

    ABSTRACT We have previously shown that multifunctional calreticulin (CRT), which resides in the endoplasmic reticulum (ER) and is involved in ER-associated protein processing, responds to infection with white spot syndrome virus (WSSV) by increasing mRNA and protein expression and by forming a complex with gC1qR and thereby delaying apoptosis. Here, we show that CRT can directly interact with WSSV structural proteins, including VP15 and VP28, during an early stage of virus infection. The binding of VP28 with CRT does not promote WSSV entry, and CRT-VP15 interaction was detected in the viral genome in virally infected host cells and thus may have an effect on WSSV replication. Moreover, CRT was detected in the viral envelope of purified WSSV virions. CRT was also found to be of high importance for proper oligomerization of the viral structural proteins VP26 and VP28, and when CRT glycosylation was blocked with tunicamycin, a significant decrease in both viral replication and assembly was detected. Together, these findings suggest that CRT confers several advantages to WSSV, from the initial steps of WSSV infection to the assembly of virions. Therefore, CRT is required as a “vital factor” and is hijacked by WSSV for its replication cycle. IMPORTANCE White spot syndrome virus (WSSV) is a double-stranded DNA virus and the cause of a serious disease in a wide range of crustaceans that often leads to high mortality rates. We have previously shown that the protein calreticulin (CRT), which resides in the endoplasmic reticulum (ER) of the cell, is important in the host response to the virus. In this report, we show that the virus uses this host protein to enter the cell and to make the host produce new viral structural proteins. Through its interaction with two viral proteins, the virus “hijacks” host calreticulin and uses it for its own needs. These findings provide new insight into the interaction between a large DNA virus and the host protein CRT and may help in understanding the viral infection process in general. PMID:24807724

  9. Analysis of a genomic segment of white spot syndrome virus of shrimp containing ribonucleotide reductase genes and repeat regions

    OpenAIRE

    Hulten, M.C.W., van; Tsai, M.F.; Schipper, C.A.; Lo, C F; Kou, G.H.; J. M. Vlak

    2000-01-01

    White spot syndrome is a worldwide disease of penaeid shrimp. The disease agent is a bacilliform, enveloped virus, white spot syndrome virus (WSSV), with a double-stranded DNA genome that probably contains well over 200 kb. Analysis of a 12?3 kb segment of WSSV DNA revealed eight open reading frames (ORFs), including the genes for the large (RR1) and small (RR2) subunits of ribonucleotide reductase. The rr1 and rr2 genes were separated by 5760 bp, containing several putative ORFs and two doma...

  10. Identification and characterization of a new E3 ubiquitin ligase in white spot syndrome virus involved in virus latency

    OpenAIRE

    Kwang Jimmy; He Fang

    2008-01-01

    Abstract White spot syndrome virus (WSSV) is one major pathogen in shrimp aquaculture. WSSV ORF403 is predicted to encode a protein of 641 amino acids, which contains a C3H2C2 RING structure. In the presence of an E2 conjugating enzyme from shrimp, WSSV403 can ubiquitinate itself in vitro, indicating it can function as a viral E3 ligase. Besides, WSSV403 E3 ligase can be activated by a series of E2 variants. Based on RT-PCR and Real time PCR, we detected transcription of WSSV403 in the commer...

  11. Comprehensive characterization of viral miRNAs involved in white spot syndrome virus (WSSV) infection.

    Science.gov (United States)

    He, Yaodong; Zhang, Xiaobo

    2012-07-01

    Guided by miRNAs, RNAi plays an important role in virus-host interactions by fine-tuning gene expression. Many viral and cellular miRNAs are involved in virus infection, though no comprehensive general model for miRNAs derived from invertebrate DNA viruses exists for their function in eukaryotic systems, despite extensive research on miRNAs. To address this issue, the miRNAs from shrimp white spot syndrome virus (WSSV), a DNA virus with a 305 kb double-stranded circular DNA genome, were characterized. Based on WSSV miRNA microarray and northern blot analyses, WSSV was shown to possess the capacity to encode 40 distinct viral miRNAs, a miRNA content roughly 360 times greater than that of humans. These findings suggested that the high content of viral miRNAs might greatly contribute to viral variability in response selective pressures in the host environment. Transcription analysis revealed that 80% of WSSV miRNAs were expressed during early stages of viral infection, indicating their importance in initial infective processes. Additionally, biogenesis of viral miRNAs was demonstrated to be dependent on host Drosha and Dicer 1, mediated by Ago 1, and viral miRNAs, including WSSV-miR211 and WSSV-miR212, were required for successful WSSV infection. During WSSV infection, numerous viral genes were likely targeted by WSSV miRNAs. The current study presented the first comprehensive view of viral miRNAs encoded by an invertebrate DNA virus, providing insight into the molecular events of virus-host interactions. PMID:22832246

  12. Silencing shrimp white spot syndrome virus (WSSV) genes by siRNA.

    Science.gov (United States)

    Xu, Jianyang; Han, Fang; Zhang, Xiaobo

    2007-02-01

    White spot syndrome virus (WSSV) is a major shrimp pathogen causing large economic losses all over the world. So far, however, there is no efficient approach to control this virus. RNA interference (RNAi), which has been applied to silence virus genes in eukaryotic organisms. In this investigation, a specific 21bp short interfering RNA (vp28-siRNA) targeting a major envelope protein gene (vp28) of WSSV was used to induce gene silencing in vivo in Penaeus japonicus shrimp. It was found that the transcription and expression of vp28 gene were silenced by the sequence-specific vp28-siRNA. However, the RNAi effect disappeared or significantly weakened even if one-nucleotide mutation existed in the vp28-siRNA. As revealed by quantitative PCR, the vp28-siRNA caused a significant reduction in viral DNA production of WSSV-infected shrimp. When treated with the vp28-siRNA, WSSV-infected shrimp had a reduced mortality rate. After three injections of the vp28-siRNA, the virus was completely eradicated from WSSV-infected shrimp. These findings suggest that RNAi is capable of silencing sequence-specific genes of WSSV and might constitute a new therapeutic strategy for WSSV infection in shrimp. PMID:17011052

  13. Detection and Quantification of Infectious Hypodermal and Hematopoietic Necrosis Virus and White Spot Virus in Shrimp Using Real-Time Quantitative PCR and SYBR Green Chemistry

    OpenAIRE

    Dhar, Arun K.; Roux, Michelle M.; Klimpel, Kurt R.

    2001-01-01

    A rapid and highly sensitive real-time PCR detection and quantification method for infectious hypodermal and hematopoietic necrosis virus (IHHNV), a single-stranded DNA virus, and white spot virus (WSV), a double-stranded DNA (dsDNA) virus infecting penaeid shrimp (Penaeus sp.), was developed using the GeneAmp 5700 sequence detection system coupled with SYBR Green chemistry. The PCR mixture contains a fluorescence dye, SYBR Green, which upon binding to dsDNA exhibits fluorescence enhancement....

  14. First report of White spot syndrome virus in farmed and wild penaeid shrimp from lagoa dos patos estuary, southern brazil

    OpenAIRE

    Cavalli, Lissandra Souto; Romano, Luis Alberto; Marins, Luis Fernando; Abreu, Paulo César

    2011-01-01

    In this study, we detected White spot syndrome virus (WSSV) in wild Farfantepenaeus paulensis collected in the Lagoa dos Patos estuary and cultivated Litopenaeus vannamei. This is the first report of WSSV in F. paulensis from Lagoa dos Patos and farmed L. vannamei shrimps in Rio Grande do Sul.

  15. First report of White spot syndrome virus in farmed and wild penaeid shrimp from Lagoa dos Patos estuary, southern Brazil

    OpenAIRE

    Lissandra Souto Cavalli; Luis Alberto Romano; Luis Fernando Marins; Paulo César Abreu

    2011-01-01

    In this study, we detected White spot syndrome virus (WSSV) in wild Farfantepenaeus paulensis collected in the Lagoa dos Patos estuary and cultivated Litopenaeus vannamei. This is the first report of WSSV in F. paulensis from Lagoa dos Patos and farmed L. vannamei shrimps in Rio Grande do Sul.

  16. First report of White spot syndrome virus in farmed and wild penaeid shrimp from Lagoa dos Patos estuary, southern Brazil

    Directory of Open Access Journals (Sweden)

    Lissandra Souto Cavalli

    2011-09-01

    Full Text Available In this study, we detected White spot syndrome virus (WSSV in wild Farfantepenaeus paulensis collected in the Lagoa dos Patos estuary and cultivated Litopenaeus vannamei. This is the first report of WSSV in F. paulensis from Lagoa dos Patos and farmed L. vannamei shrimps in Rio Grande do Sul.

  17. First report of White spot syndrome virus in farmed and wild penaeid shrimp from Lagoa dos Patos estuary, southern Brazil

    Scientific Electronic Library Online (English)

    Lissandra Souto, Cavalli; Luis Alberto, Romano; Luis Fernando, Marins; Paulo César, Abreu.

    2011-09-01

    Full Text Available In this study, we detected White spot syndrome virus (WSSV) in wild Farfantepenaeus paulensis collected in the Lagoa dos Patos estuary and cultivated Litopenaeus vannamei. This is the first report of WSSV in F. paulensis from Lagoa dos Patos and farmed L. vannamei shrimps in Rio Grande do Sul. [...

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

    Science.gov (United States)

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

    2015-05-01

    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 et al. 2013); therefore, it is a potential propagative vector for virus transmission. The major aim of this study was to determine whether WSSV can be transmitted from naturally infected Dendronereis spp. to specific pathogen-free (SPF) Pacific white shrimp Litopenaeus vannamei (Boone) through feeding. WSSV was detected in naturally infected Dendronereis spp. and Penaeus monodon Fabricius from a traditional shrimp pond, and the positive animals were used in the current experiment. WSSV-infected Dendronereis spp. and P. monodon in a pond had a point prevalence of 90% and 80%, respectively, as measured by PCR. WSSV was detected in the head, gills, blood and mid-body of Dendronereis spp. WSSV from naturally infected Dendronereis spp was transmitted to SPF L. vannamei and subsequently from this shrimp to new naïve-SPF L. vannamei to cause transient infection. Our findings support the contention that Dendronereis spp, upon feeding, can be a source of WSSV infection of shrimp in ponds. PMID:24716813

  19. Functional identification of the non-specific nuclease from white spot syndrome virus

    International Nuclear Information System (INIS)

    The product encoded by the wsv191 gene from shrimp white spot syndrome virus (WSSV) is homologous with non-specific nucleases (NSN) of other organisms. To functionally identify the protein, the wsv191 gene was expressed in Escherichia coli as a glutathione S-transferase (GST) fusion protein with 6His-tag at C-terminal. The fusion protein (termed as rWSSV-NSN) was purified using Ni-NTA affinity chromatography under denatured conditions, renatured and characterized by three methods. The results showed that rWSSV-NSN could hydrolyze both DNA and RNA. 5'-RACE result revealed that the transcription initiation site of the wsv191 gene was located at nucleotide residue G of the predicted ATG triplet. Therefore, we concluded that the next ATG should be the genuine translation initiation codon of the wsv191 gene. Western blot analysis revealed that the molecular mass of natural WSSV-NSN was 37 kDa

  20. Development of primers for loop-mediated isothermal amplification of Philippine white spot syndrome virus isolates

    Directory of Open Access Journals (Sweden)

    Benedict A. Maralit

    2012-09-01

    Full Text Available Because there are currently no effective chemicals or drugs to completely treat bacterial andviral diseases, effective means of prevention such as early detection methods are continuously beingexplored and fully exhausted. Over the past few years, we have seen the development of cutting edgetechnologies to specifically and efficiently detect White Spot Syndrome Virus (WSSV, one of the majorviruses that devastated global shrimp aquaculture. The recent discovery of loop mediated isothermalamplification (LAMP brought about opportunities for fast diagnosis of a lot of animal diseases includinghumans. Here, the development of alternative LAMP primers specifically for Philippine WSSV isolates wasdiscussed. The sensitivity of the established detection protocol was found to be at 0.3954 pg of shrimpDNA template while exhibiting high specificity to the viral target. In addition, alternative visualizationtechniques and comparison with other detection protocols are also discussed.

  1. Characterization of ORF89 - A latency-related gene of white spot syndrome virus

    International Nuclear Information System (INIS)

    Open reading frame 89 (ORF89) is one of the three genes that are believed to be involved in the latent infection of white spot syndrome virus (WSSV). Here, we report the structure and functional characterization of ORF89. cDNA sequencing, 5' RLM-RACE, and 3' RLM-RACE showed that ORF89 gene is transcribed into an unspliced mRNA of 4436 nucleotides, which is predicted to encode a protein of 1437 amino acids. ORF89 expressed an approximately 165-kDa protein in Sf9 cells that localized in the nucleus. Amino acids 678-683 were found to be essential for nuclear localization. Cotransfection assays demonstrated that ORF89 protein repressed its own promoter as well as those of a protein kinase and the thymidine-thymidylate kinase genes of WSSV. SYBR Green real-time PCR indicated that the repression occurred at the transcriptional level

  2. Genotyping of white spot syndrome virus on wild and farm crustaceans from Sonora, Mexico

    Directory of Open Access Journals (Sweden)

    González-Galaviz José Reyes

    2013-01-01

    Full Text Available White spot syndrome is a viral disease affecting wild and farm crustaceans that serve as reservoirs. Previous reports have demonstrated high genomic variation in WSS viruses (WSSV isolated from distinct geographical regions. In this study, we collected wild shrimps (Litopenaeus stylirostris, crabs (Callinectes arcuatus and farmed shrimp (L. vannamei in Sonora, Mexico, between 2008 and 2010. DNA was extracted, and the variable regions and transposase genes were subjected to PCR and sequencing. Compared to strains of WSSV from other sites, Mexican samples exhibited a distinct number of repeat units (RUs in ORF94, ORF75 and ORF125, which ranged between 1-11, 3-15, and 8-11 RUs respectively, and a unique single nucleotide polymorphism (SNP at position 48 of ORF94. A total of six Mexican genotypes were found in organism from shrimp farm and natural environment.

  3. Characterization of white spot syndrome virus VP52B and its interaction with VP26.

    Science.gov (United States)

    Lin, Fanyu; Jie, Zuliang; Hou, Luhong; Li, Fang; Yang, Feng

    2015-02-01

    White spot syndrome virus (WSSV) is one of the major pathogens of cultured shrimp. Identification of envelope protein interactions has become a central issue for the understanding of WSSV assembly. In this paper, WSSV envelope protein VP52B was fused with GST-tag and expressed in Escherichia coli BL-21(DE3). Immunogold-electron microscopy revealed that VP52B was located on the outside surface of WSSV virions. Far-Western blotting analysis suggested that VP52B might directly interact with a major viral envelope protein VP26, and their interaction was confirmed by GST pull-down assay. Further investigation showed that the VP52B binding domain was located between residues 135-170 of VP26. These findings will enhance our understanding of the molecular mechanisms of WSSV morphogenesis. PMID:25331340

  4. Expression, purification and crystallization of a novel nonstructural protein VP9 from white spot syndrome virus

    Science.gov (United States)

    Liu, Yang; Sivaraman, J.; Hew, Choy L.

    2006-01-01

    The nonstructural protein VP9 from white spot syndrome virus (WSSV) has been identified and expressed in Escherichia coli. To facilitate purification, a cleavable His6 tag was introduced at the N-terminus. The native protein was purified and crystallized by vapour diffusion against mother liquor containing 2?M sodium acetate, 100?mM MES pH 6.3, 25?mM cadmium sulfate and 3% glycerol. Crystals were obtained within 7?d and diffracted to 2.2?Å; they belonged to space group P212121, with unit-cell parameters a = 74.13, b = 78.21, c = 78.98?Å and four molecules in the asymmetric unit. The selenomethionine-labelled protein produced isomorphous crystals that diffracted to approximately 3.3?Å. PMID:16880562

  5. White Spot Syndrome Virus Induces Metabolic Changes Resembling the Warburg Effect in Shrimp Hemocytes in the Early Stage of Infection ?

    OpenAIRE

    Chen, I-Tung; Aoki, Takashi; Huang, Yun-Tzu; Hirono, Ikuo; Chen, Tsan-Chi; Huang, Jiun-Yan; Chang, Geen-Dong; Lo, Chu-Fang; Wang, Han-Ching

    2011-01-01

    The Warburg effect is an abnormal glycolysis response that is associated with cancer cells. Here we present evidence that metabolic changes resembling the Warburg effect are induced by a nonmammalian virus. When shrimp were infected with white spot syndrome virus (WSSV), changes were induced in several metabolic pathways related to the mitochondria. At the viral genome replication stage (12 h postinfection [hpi]), glucose consumption and plasma lactate concentration were both increased in WSS...

  6. Identification and phylogeny of a non-specific nuclease gene of white spot syndrome virus of shrimp

    OpenAIRE

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

    2001-01-01

    White spot syndrome virus (WSSV) is a taxonomically unclassified virus which causes a disease in shrimps worldwide. A 936 bp long open reading frame (ORF) was found on a 7.2 kb HindIII fragment of the DNA genome of WSSV located adjacent to the ribonucleotide reductase small subunit gene. This putative ORF showed homology to prokaryotic and eukaryotic endonucleases, which contain a non-specific endonuclease motif. Alignment with viral and eukaryotic endonuclease ORFs revealed that most catalyt...

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

    OpenAIRE

    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 water parameters on a farm with 11 ponds that had WSSV infection. The virus was detected in shrimp exposed to two or more simultaneous stress factors (temperature, pH, water level) or multiple stres...

  8. Mixed-genotype white spot syndrome virus infections of shrimp are inversely correlated with disease outbreaks in ponds

    OpenAIRE

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

    2011-01-01

    Outbreaks of white spot syndrome virus (WSSV) in shrimp culture and its relation to virus virulence are not well understood. Here we provide evidence that the presence of WSSV mixed-genotype infections correlate with lower outbreak incidence and that disease outbreaks correlate with single-genotype infections. We tested 573 shrimp samples from 81 shrimp ponds in the Mekong delta with outbreak or non-outbreak status. The variable number tandem repeat (VNTR) loci of WSSV were used as molecular ...

  9. A Putative Cell Surface Receptor for White Spot Syndrome Virus Is a Member of a Transporter Superfamily

    OpenAIRE

    Huang, Huai-Ting; Leu, Jiann-Horng; Huang, Po-Yu; Li-li CHEN

    2012-01-01

    White spot syndrome virus (WSSV), a large enveloped DNA virus, can cause the most serious viral disease in shrimp and has a wide host range among crustaceans. In this study, we identified a surface protein, named glucose transporter 1 (Glut1), which could also interact with WSSV envelope protein, VP53A. Sequence analysis revealed that Glut1 is a member of a large superfamily of transporters and that it is most closely related to evolutionary branches of this superfamily, branches that functio...

  10. Transcript Analysis of White spot syndrome virus Latency and Phagocytosis Activating Protein Genes in Infected Shrimp (Penaeus monodon)

    OpenAIRE

    Shekhar, M. S.; DilliKumar, M.; Vinaya Kumar, K.; G. Gopikrishna; Rajesh, S; Kiruthika, J.; Ponniah, A. G.

    2012-01-01

    Viral latency has been recently observed to be associated with White spot syndrome virus (WSSV) infection in shrimp. In the present study, shrimp samples (Penaeus monodon) surviving WSSV infection were examined for presence of WSSV in latent phase. Virus latency was observed in shrimp which were either experimentally challenged with WSSV and survived the infection or those which survived the natural infection. Three viral transcripts (ORFs 427, 151, 366) associated with latency were analyzed ...

  11. Baculovirus-mediated promoter assay and transcriptional analysis of white spot syndrome virus orf427 gene

    Directory of Open Access Journals (Sweden)

    Yu Li

    2005-08-01

    Full Text Available Abstract Background White spot syndrome virus (WSSV is an important pathogen of the penaeid shrimp with high mortalities. In previous reports, Orf427 of WSSV is characterized as one of the three major latency-associated genes of WSSV. Here, we were interested to analyze the promoter of orf427 and its expression during viral pathogenesis. Results in situ hybridization revealed that orf427 was transcribed in all the infected tissues during viral lytic infection and the translational product can be detected from the infected shrimp. A time-course RT-PCR analysis indicated that transcriptional products of orf427 could only be detected after 6 h post virus inoculation. Furthermore, a baculovirus-mediated promoter analysis indicated that the promoter of orf427 failed to express the EGFP reporter gene in both insect SF9 cells and primary shrimp cells. Conclusion Our data suggested that latency-related orf427 might not play an important role in activating virus replication from latent phase due to its late transcription during the lytic infection.

  12. Proteomic Analysis of Differentially Expressed Proteins in Fenneropenaeus chinensis Hemocytes upon White Spot Syndrome Virus Infection

    Science.gov (United States)

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

    2014-01-01

    To elucidate molecular responses of shrimp hemocytes to white spot syndrome virus (WSSV) infection, two-dimensional gel electrophoresis was applied to investigate differentially expressed proteins in hemocytes of Chinese shrimp (Fenneropenaeus chinensis) at 24 h post infection (hpi). Approximately 580 protein spots were detected in hemocytes of healthy and WSSV-infected shrimps. Quantitative intensity analysis revealed 26 protein spots were significantly up-regulated, and 19 spots were significantly down-regulated. By mass spectrometry, small ubiquitin-like modifier (SUMO) 1, cytosolic MnSOD, triosephosphate isomerase, tubulin alpha-1 chain, microtubule-actin cross-linking factor 1, nuclear receptor E75 protein, vacuolar ATP synthase subunit B L form, inositol 1,4,5-trisphosphate receptor, arginine kinase, etc., amounting to 33 differentially modulated proteins were identified successfully. According to Gene Ontology annotation, the identified proteins were classified into nine categories, consisting of immune related proteins, stimulus response proteins, proteins involved in glucose metabolic process, cytoskeleton proteins, DNA or protein binding proteins, proteins involved in steroid hormone mediated signal pathway, ATP synthases, proteins involved in transmembrane transport and ungrouped proteins. Meanwhile, the expression profiles of three up-regulated proteins (SUMO, heat shock protein 70, and arginine kinase) and one down-regulated protein (prophenoloxidase) were further analyzed by real-time RT-PCR at the transcription level after WSSV infection. The results showed that SUMO and heat shock protein 70 were significantly up-regulated at each sampling time point, while arginine kinase was significantly up-regulated at 12 and 24 hpi. In contrast, prophenoloxidase was significantly down-regulated at each sampling time point. The results of this work provided preliminary data on proteins in shrimp hemocytes involved in WSSV infection. PMID:24587154

  13. Tangential flow ultrafiltration for detection of white spot syndrome virus (WSSV) in shrimp pond water.

    Science.gov (United States)

    Alavandi, S V; Ananda Bharathi, R; Satheesh Kumar, S; Dineshkumar, N; Saravanakumar, C; Joseph Sahaya Rajan, J

    2015-06-15

    Water represents the most important component in the white spot syndrome virus (WSSV) transmission pathway in aquaculture, yet there is very little information. Detection of viruses in water is a challenge, since their counts will often be too low to be detected by available methods such as polymerase chain reaction (PCR). In order to overcome this difficulty, viruses in water have to be concentrated from large volumes of water prior to detection. In this study, a total of 19 water samples from aquaculture ecosystem comprising 3 creeks, 10 shrimp culture ponds, 3 shrimp broodstock tanks and 2 larval rearing tanks of shrimp hatcheries and a sample from a hatchery effluent treatment tank were subjected to concentration of viruses by ultrafiltration (UF) using tangential flow filtration (TFF). Twenty to 100l of water from these sources was concentrated to a final volume of 100mL (200-1000 fold). The efficiency of recovery of WSSV by TFF ranged from 7.5 to 89.61%. WSSV could be successfully detected by PCR in the viral concentrates obtained from water samples of three shrimp culture ponds, one each of the shrimp broodstock tank, larval rearing tank, and the shrimp hatchery effluent treatment tank with WSSV copy numbers ranging from 6 to 157mL(-1) by quantitative real time PCR. The ultrafiltration virus concentration technique enables efficient detection of shrimp viral pathogens in water from aquaculture facilities. It could be used as an important tool to understand the efficacy of biosecurity protocols adopted in the aquaculture facility and to carry out epidemiological investigations of aquatic viral pathogens. PMID:25779823

  14. Immune response of black tiger shrimp (Penaeus monodon Fabricius to yellow head virus (YHV and white spot syndrome virus (WSSV

    Directory of Open Access Journals (Sweden)

    Supamattaya, K.

    2005-02-01

    Full Text Available Immunological responses of black tiger shrimp (Penaeus monodon were induced under laboratory conditions by injecting white spot syndrome virus (WSSV and yellow head virus (YHV. The survivors from WSSV and YHV infection showed improvement in their immunity against the re-challenging by both viruses. The WSSV survivors showed higher capability than that of YHV survivors in developing the immunity. The highest relative percent survival (RPS against WSSV and YHV noted after a 43-day period of WSSV injection was 67 and 37.5%, respectively. While the RPSs against both viruses after periods of 46 and 60 days of the YHV injection were 37.5 and 12.1%. Similarly, an in vitro neutralization activity of plasma separated from the survivors with WSSV injection showed preference of virus being eliminated over the plasma from YHV injection.Blood parameters for survivors with WSSV injection for 43 days showed an increase in phenoloxidase activity, while the YHV injected survivors exhibited higher level of total hemocytes and phenoloxidase activity. Histopathological examinations in survivors revealed changes of lymphatic tubes into spheroids higher than those in normal individual.

  15. In silico identification of putative promoter motifs of White Spot Syndrome Virus

    Directory of Open Access Journals (Sweden)

    Sandbrink Hans

    2006-06-01

    Full Text Available Abstract 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 cascaded fashion. To search in silico for conserved promoter motifs (i the abundance of all 4 through 8 nucleotide motifs in the upstream sequences of WSSV genes relative to the complete genome was determined, and (ii a MEME search was performed in the upstream sequences of either early or late WSSV genes, as assigned by microarray analysis. Both methods were validated by alignments of empirically determined 5' ends of various WSSV mRNAs. Results The collective information shows that the upstream region of early WSSV genes, containing a TATA box and an initiator, is similar to Drosophila RNA polymerase II core promoter sequences, suggesting utilization of the cellular transcription machinery for generating early transcripts. The alignment of the 5' ends of known well-established late genes, including all major structural protein genes, identified a degenerate motif (ATNAC which could be involved in WSSV late transcription. For these genes, only one contained a functional TATA box. However, almost half of the WSSV late genes, as previously assigned by microarray analysis, did contain a TATA box in their upstream region. Conclusion The data may suggest the presence of two separate classes of late WSSV genes, one exploiting the cellular RNA polymerase II system for mRNA synthesis and the other generating messengers by a new virus-induced transcription mechanism.

  16. Characterization and Interactome Study of White Spot Syndrome Virus Envelope Protein VP11

    Science.gov (United States)

    Liu, Wang-Jing; Shiung, Hui-Jui; Lo, Chu-Fang; Leu, Jiann-Horng; Lai, Ying-Jang; Lee, Tai-Lin; Huang, Wei-Tung; Kou, Guang-Hsiung; Chang, Yun-Shiang

    2014-01-01

    White spot syndrome virus (WSSV) is a large enveloped virus. The WSSV viral particle consists of three structural layers that surround its core DNA: an outer envelope, a tegument and a nucleocapsid. Here we characterize the WSSV structural protein VP11 (WSSV394, GenBank accession number AF440570), and use an interactome approach to analyze the possible associations between this protein and an array of other WSSV and host proteins. Temporal transcription analysis showed that vp11 is an early gene. Western blot hybridization of the intact viral particles and fractionation of the viral components, and immunoelectron microscopy showed that VP11 is an envelope protein. Membrane topology software predicted VP11 to be a type of transmembrane protein with a highly hydrophobic transmembrane domain at its N-terminal. Based on an immunofluorescence assay performed on VP11-transfected Sf9 cells and a trypsin digestion analysis of the virion, we conclude that, contrary to topology software prediction, the C-terminal of this protein is in fact inside the virion. Yeast two-hybrid screening combined with co-immunoprecipitation assays found that VP11 directly interacted with at least 12 other WSSV structural proteins as well as itself. An oligomerization assay further showed that VP11 could form dimers. VP11 is also the first reported WSSV structural protein to interact with the major nucleocapsid protein VP664. PMID:24465701

  17. Crayfish hematopoietic tissue cells but not hemocytes are permissive for white spot syndrome virus replication.

    Science.gov (United States)

    Wu, Junjun; Li, Fang; Huang, Jiajun; Xu, Limei; Yang, Feng

    2015-03-01

    Hemocytes are the major immune cells of crustaceans which are believed to be essential for the pathogenesis of white spot syndrome virus (WSSV) infection. Crayfish hemocytes and hematopoietic tissue (HPT) cells have been found to be susceptible to WSSV infection, but the procedure of WSSV infection to both cell types has not yet been carefully investigated. In this study, we analyzed the infection and proliferation of WSSV in crayfish hemocytes as well as HPT cells in detail through transmission electronic microscopy (TEM). The results showed that WSSV could enter both hemocytes and HPT cells through endocytosis, but the production of progeny virus was only achieved in HPT cells. Further investigation demonstrated that although WSSV could transcribe its genes in both cell types, viral genome replication and structural protein expression were unsuccessful in hemocytes, which may be responsible for the failure of progeny production. Therefore, we propose that both hemocytes and HPT cells are susceptible to WSSV infection but only HPT cells are permissive to WSSV replication. These findings will extend our knowledge of the interaction between WSSV and the host immune system. PMID:25541079

  18. Construction and Application of a Protein Interaction Map for White Spot Syndrome Virus (WSSV)*

    Science.gov (United States)

    Sangsuriya, Pakkakul; Huang, Jiun-Yan; Chu, Yu-Fei; Phiwsaiya, Kornsunee; Leekitcharoenphon, Pimlapas; Meemetta, Watcharachai; Senapin, Saengchan; Huang, Wei-Pang; Withyachumnarnkul, Boonsirm; Flegel, Timothy W.; Lo, Chu-Fang

    2014-01-01

    White spot syndrome virus (WSSV) is currently the most serious global threat for cultured shrimp production. Although its large, double-stranded DNA genome has been completely characterized, most putative protein functions remain obscure. To provide more informative knowledge about this virus, a proteomic-scale network of WSSV-WSSV protein interactions was carried out using a comprehensive yeast two-hybrid analysis. An array of yeast transformants containing each WSSV open reading frame fused with GAL4 DNA binding domain and GAL4 activation domain was constructed yielding 187 bait and 182 prey constructs, respectively. On screening of ?28,000 pairwise combinations, 710 interactions were obtained from 143 baits. An independent coimmunoprecipitation assay (co-IP) was performed to validate the selected protein interaction pairs identified from the yeast two-hybrid approach. The program Cytoscape was employed to create a WSSV protein–protein interaction (PPI) network. The topology of the WSSV PPI network was based on the Barabási-Albert model and consisted of a scale-free network that resembled other established viral protein interaction networks. Using the RNA interference approach, knocking down either of two candidate hub proteins gave shrimp more protection against WSSV than knocking down a nonhub gene. The WSSV protein interaction map established in this study provides novel guidance for further studies on shrimp viral pathogenesis, host-viral protein interaction and potential targets for therapeutic and preventative antiviral strategies in shrimp aquaculture. PMID:24217020

  19. Recent insights into host-pathogen interaction in white spot syndrome virus infected penaeid shrimp.

    Science.gov (United States)

    Shekhar, M S; Ponniah, A G

    2015-07-01

    Viral disease outbreaks are a major concern impeding the development of the shrimp aquaculture industry. The viral disease due to white spot syndrome virus (WSSV) observed in early 1990s still continues unabated affecting the shrimp farms and cause huge economic loss to the shrimp aquaculture industry. In the absence of effective therapeutics to control WSSV, it is important to understand viral pathogenesis and shrimp response to WSSV at the molecular level. Identification and molecular characterization of WSSV proteins and receptors may facilitate in designing and development of novel therapeutics and antiviral drugs that may inhibit viral replication. Investigations into host-pathogen interactions might give new insights to viral infectivity, tissue tropism and defence mechanism elicited in response to WSSV infection. However, due to the limited information on WSSV gene function and host immune response, the signalling pathways which are associated in shrimp pathogen interaction have also not been elucidated completely. In the present review, the focus is on those shrimp proteins and receptors that are potentially involved in virus infection or in the defence mechanism against WSSV. In addition, the major signalling pathways involved in the innate immune response and the role of apoptosis in host-pathogen interaction is discussed. PMID:24953507

  20. Nucleocapsid protein VP15 is the basic DNA binding protein of white spot syndrome virus of shrimp

    OpenAIRE

    Witteveldt, J.; Vermeesch, A.M.G.; Langenhof, M.; Lang, A., de; J. M. Vlak; Hulten, M.C.W., van

    2005-01-01

    White spot syndrome virus (WSSV) is type species of the genus Whispovirus of the new family Nimaviridae. Despite the elucidation of its genomic sequence, very little is known about the virus as only 6% of its ORFs show homology to known genes. One of the structural virion proteins, VP15, is part of the nucleocapsid of the virus and shows homology to some putative baculovirus DNA binding proteins. These DNA-binding or histone-like proteins are thought to be involved in the condensation and pac...

  1. PCR detection of white spot syndrome virus (WSSV) from farmed Pacific white shrimp (Litopenaeus vannamei) in selected sites of the Philippines

    OpenAIRE

    Mary Beth B. Maningas; Santos, Mudjekeewis D.; Benedict A. Maralit; Christopher M. A. Caipang

    2011-01-01

    Great losses caused by white spot syndrome virus (WSSV) in shrimp culture have beenattributed to poor screening procedures in farms and the lack of sufficient access to specific pathogenfree brood stock. Thus, early detection of the virus is considered the best option for shrimp farmers. Thestudy, thus, assessed viral incidence in the Philippines and partially sequenced and characterized thePhilippine WSSV isolate with regards to other isolates in GenBank. Developed primers for PCR can detect...

  2. Gene-expression profiling of White spot syndrome virus in vivo.

    Science.gov (United States)

    Marks, Hendrik; Vorst, Oscar; van Houwelingen, Adèle M M L; van Hulten, Mariëlle C W; Vlak, Just M

    2005-07-01

    White spot syndrome virus, type species of the genus Whispovirus in the family Nimaviridae, is a large, double-stranded DNA (dsDNA) virus that infects crustaceans. The genome of the completely sequenced isolate WSSV-TH encodes 184 putative open reading frames (ORFs), the functions of which are largely unknown. To study the transcription of these ORFs, a DNA microarray was constructed, containing probes corresponding to nearly all putative WSSV-TH ORFs. Transcripts of 79 % of these ORFs could be detected in the gills of WSSV-infected shrimp (Penaeus monodon). Clustering of the transcription profiles of the individual genes during infection showed two major classes of genes: the first class reached maximal expression at 20 h post-infection (p.i.) (putative early) and the other class at 2 days p.i. (putative late). Nearly all major and minor structural virion-protein genes clustered in the latter group. These data provide evidence that, similar to other large, dsDNA viruses, the WSSV genes at large are expressed in a coordinated and cascaded fashion. Furthermore, the transcriptomes of the WSSV isolates WSSV-TH and TH-96-II, which have differential virulence, were compared at 2 days p.i. The TH-96-II genome encodes 10 ORFs that are not present in WSSV-TH, of which at least seven were expressed in P. monodon as well as in crayfish (Astacus leptodactylus), suggesting a functional but not essential role for these genes during infection. Expression levels of most other ORFs shared by both isolates were similar. Evaluation of transcription profiles by using a genome-wide approach provides a better understanding of WSSV transcription regulation and a new tool to study WSSV gene function. PMID:15958687

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

    Directory of Open Access Journals (Sweden)

    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 susceptible than S. paramamosain. Based on our single-challenge and serial challenge results, and on previous published work showing that S. serrata is relatively unaffected by WSSV infection, we propose that susceptibility to white spot disease in the genus Scylla is species-dependent and may also be dose-history dependent. In practical terms for shrimp farmers, it means that S. olivacea and S. paramamosain may pose less threat as WSSV carriers than S. serrata. For crab farmers, our results suggest that rearing of S. serrata would be a better choice than S. paramamosain or S. olivacea in terms of avoiding losses from seasonal outbreaks of white spot disease.

  4. Evaluation of an immunodot test to manage white spot syndrome virus (WSSV) during cultivation of the giant tiger shrimp Penaeus monodon

    Digital Repository Service at National Institute of Oceanography (India)

    Patil, R.; Palaksha, K.J.; Anil, T.M.; Guruchannabasavanna; Patil, P.; Shankar, K.M.; Mohan, C.V.; Sreepada, R.A.

    2008-01-01

    A monoclonal antibody-based immunodot test was compared to a polymerase chain reaction (PCR) assay for managing white spot syndrome virus (WSSV) on shrimp farms at Kundapur and Kumta situated in Udupi and Uttar Kannada Districts, respectively...

  5. Antilipopolysaccharide Factor Interferes with White Spot Syndrome Virus Replication In Vitro and In Vivo in the Crayfish Pacifastacus leniusculus

    OpenAIRE

    Liu, Haipeng; Jiravanichpaisal, Pikul; Söderhäll, Irene; Cerenius, Lage; Söderhäll, Kenneth

    2006-01-01

    In a study of genes expressed differentially in the freshwater crayfish Pacifastacus leniusculus infected experimentally with the white spot syndrome virus (WSSV), one protein, known as antilipopolysaccharide factor (ALF), was chosen, among those whose transcript levels increased upon viral infection, for further studies. ALF RNA interference (RNAi) experiments in whole animals and in cell cultures indicated that ALF can protect against WSSV infection, since knockdown of ALF by RNAi specifica...

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

    OpenAIRE

    Selvam, Deborah Gnana; Mujeeb Rahiman, K. M.; Mohamed Hatha, A. A.

    2012-01-01

    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 oxyg...

  7. Preparation of transgenic Dunaliella salina for immunization against white spot syndrome virus in crayfish.

    Science.gov (United States)

    Feng, Shuying; Feng, Wenpo; Zhao, Ling; Gu, Huihui; Li, Qinghua; Shi, Ke; Guo, Sanxing; Zhang, Nannan

    2014-03-01

    Although a white spot syndrome virus (WSSV) subunit vaccine could significantly enhance the immune response and benefit the shrimp host, its practical application is currently not feasible because of drawbacks in existing expression systems. We generated a transgenic Dunaliella salina (D. salina) strain by introducing the WSSV VP28 gene to produce a novel oral WSSV subunit vaccine. Following transformation of D. salina, VP28 gene expression was assessed by reverse transcription polymerase chain reaction (RT-PCR) assays, enzyme-linked immunosorbent assays (ELISAs), and western blot analysis. The RT-PCR results indicated that the VP28 gene was successfully expressed in D. salina cells. The presence of recombinant VP28 proteins with natural bioactivity was confirmed by western blot analysis and ELISA. Animal vaccination experiments indicated that transgenic D. salina can induce protection against WSSV by oral delivery in crayfish. Our findings indicate that the VP28 gene can be successfully expressed in transgenic D. salina and can be applied as an oral vaccine to protect crayfish against WSSV. We have demonstrated that it is feasible to produce an oral vaccine using D. salina, and thereby provide a new method for controlling other viral diseases in crustaceans. PMID:24081826

  8. Localization studies of two white spot syndrome virus structural proteins VP51 and VP76

    Directory of Open Access Journals (Sweden)

    Yang Feng

    2006-09-01

    Full Text Available Abstract VP51 and VP76 are two structural proteins of white spot syndrome virus (WSSV. However, there is some controversy about their localization in the virion at present. In this study, we employ multiple approaches to reevaluate the location of VP51 and VP76. Firstly, we found VP51 and VP76 presence in viral nucleocapsids fraction by Western blotting. Secondly, after the high-salt treatment of nucleocapsids, VP51 and VP76 were still exclusively present in viral capsids by Western blotting and immunoelectron microscopy, suggesting two proteins are structural components of the viral capsid. To gather more evidence, we developed a method based on immunofluorescence flow cytometry. The results revealed that the mean fluorescence intensity of the viral capsids group was significantly higher than that of intact virions group after incubation with anti-VP51 or anti-VP76 serum and fluorescein isothiocyanate conjugated secondary antibody. All these results indicate that VP51 and VP76 are both capsid proteins of WSSV.

  9. Hematological changes in white spot syndrome virus-infected shrimp, Fenneropenaeus chinensis (Osbeck)

    Science.gov (United States)

    Feng, Shouming; Zhan, Wenbin; Xing, Jing; Li, Jun; Yang, Kai; Wang, Jing

    2008-08-01

    The pathological changes of hemocytes in the haemolymph and hepatopancreas were examined in experimentally and naturally WSSV (white spot syndrome virus) infected Fenneropenaeus chinensis. The results showed that the pathological manifestations of hemocytes were similar among moribund shrimps infected via injection, feeding and by nature. Firstly, the total hemocyte counts (THCs) in WSSV-infected shrimp were significantly lower than those in healthy shrimp. Secondly, necrotic, broken and disintegrated cells were often observed, and a typical hematolysis was present in the haemolymph smear of WSSV-infected shrimp. Thirdly, necrosis and typical apoptosis of hemocytes were detected with TEM in the peripheral haemolymph of WSSV-infected shrimp. Hyalinocytes and semi-granulocytes with masses of WSSVs in their nuclei often appeared, whereas no granular hemocytes with WSSV were found in the hepatopancreas of moribund infected shrimps. All our results supported that hemocytes were the main target cells of WSSV, and hyalinocytes and semigranular hemocytes seemed to be more favorable for WSSV infection in F. chinensis.

  10. Screening, isolation and optimization of anti–white spot syndrome virus drug derived from marine plants

    Science.gov (United States)

    Chakraborty, Somnath; Ghosh, Upasana; Balasubramanian, Thangavel; Das, Punyabrata

    2014-01-01

    Objective To screen, isolate and optimize anti-white spot syndrome virus (WSSV) drug derived from various marine floral ecosystems and to evaluate the efficacy of the same in host–pathogen interaction model. Methods Thirty species of marine plants were subjected to Soxhlet extraction using water, ethanol, methanol and hexane as solvents. The 120 plant isolates thus obtained were screened for their in vivo anti-WSSV property in Litopenaeus vannamei. By means of chemical processes, the purified anti-WSSV plant isolate, MP07X was derived. The drug was optimized at various concentrations. Viral and immune genes were analysed using reverse transcriptase PCR to confirm the potency of the drug. Results Nine plant isolates exhibited significant survivability in host. The drug MP07X thus formulated showing 85% survivability in host. The surviving shrimps were nested PCR negative at the end of the 15 d experimentation. The lowest concentration of MP07X required intramuscularly for virucidal property was 10 mg/mL. The oral dosage of 1?000 mg/kg body weight/day survived at the rate of 85%. Neither VP28 nor ie 1 was expressed in the test samples at 42nd hour and 84th hour post viral infection. Conclusions The drug MP07X derived from Rhizophora mucronata is a potent anti-WSSV drug. PMID:25183065

  11. The tidepool shrimp, Palaemon ritteri Holmes, constitutes a novel host to the white spot syndrome virus.

    Science.gov (United States)

    Sánchez-Paz, A; Terán-Díaz, B; Enríquez-Espinoza, T; Encinas-Garcia, T; Vázquez-Sánchez, I; Mendoza-Cano, F

    2015-07-01

    The white spot syndrome virus (WSSV) is a lethal and contagious pathogen for penaeid shrimp and a growing number of other crustacean species. To date, there are no effective prophylactic or therapeutic treatments commercially available to interfere with the occurrence and spread of the disease. In addition, the significance of alternative vectors on the dispersal of this disease has been largely ignored and therefore the ecological dynamics of the WSSV is still poorly understood and difficult to ascertain. Thus, an important issue that should be considered in sanitary programmes and management strategies is the identification of species susceptible to infection by WSSV. The results obtained provide the first direct evidence of ongoing WSSV replication in experimentally infected specimens of the tidepool shrimp Palaemon ritteri. Viral replication was detected using a validated set of primers for the amplification by RT-PCR of a 141 bp fragment of the transcript encoding the viral protein VP28. It is therefore conceivable that this shrimp may play a significant role in the dispersal of WSSV. PMID:24953350

  12. Oral Vaccination of Baculovirus-Expressed VP28 Displays Enhanced Protection against White Spot Syndrome Virus in Penaeus monodon

    OpenAIRE

    S, Syed Musthaq; Kwang, Jimmy

    2011-01-01

    White Spot Syndrome Virus (WSSV) is an infectious pathogen of shrimp and other crustaceans, and neither effective vaccines nor adequate treatments are currently available. WSSV is an enveloped dsDNA virus, and one of its major envelope proteins, VP28, plays a pivotal role in WSSV infection. In an attempt to develop a vaccine against WSSV, we inserted the VP28 gene into a baculovirus vector tailored to express VP28 on the baculovirus surface under the WSSV ie1 promoter (Bac-VP28). The Bac-VP28...

  13. Effects of Water Temperature on the White Spot Syndrome Virus Infection in Postlarvae Litopenaeus vannamei

    Directory of Open Access Journals (Sweden)

    Sutee WONGMANEEPRATEEP

    2010-06-01

    Full Text Available This study evaluated the effects of high water temperature (32 ± 1 °C on the white spot syndrome virus (WSSV infection in Litopenaeus vannamei postlarvae (PL15. WSSV challenge was done by immersion. One group of PL15 was continuously maintained at 32 ± 1 °C until the end of the experiment after challenge and a control group of PL15 was constantly maintained at 28 ± 1 °C until the end of the experiment after challenge. Other groups were kept at 32 ± 1 °C until temperature was altered from 32 ± 1 °C to 28 ± 1 °C at 0, 1, 3, 5 and 7 days after infection. Gross signs and mortality were monitored every 12 h until the end of the experiment. WSSV infections were confirmed by nested-PCR, histopathology, immunohistochemistry and bioassay methods. Challenged shrimp were kept at 32 ± 1 °C for 0, 1, 3 and 5 days before the temperature was reduced to 28 ± 1 °C revealing that maintaining the temperature at 32 ± 1 °C for a longer period could delay clinical signs and onset of mortalities. Nevertheless, 100 % mortalities occurred in all groups and the control group within 7 days. All moribund PL15 were WSSV-positive by nested-PCR assay as well as histopathology, immunohistochemistry and bioassay methods. In contrast, PL15 constantly maintained at 32 ± 1 °C until the end of the experiment, and for 7 days after challenge before switching to 28 ± 1 °C did not show clinical signs and mortality. Surviving PL15 from both groups were WSSV-negative by nested-PCR assay as well as histopathology, immunohistochemistry and bioassay methods. This study clearly indicated that postlarvae maintained constantly at 32 ± 1 °C for 7 days were able to eliminate/clear WSSV infection.

  14. Reprint of "evolution of specific immunity in shrimp - a vaccination perspective against white spot syndrome virus".

    Science.gov (United States)

    Syed Musthaq, Syed Khader; Kwang, Jimmy

    2015-02-01

    Invertebrates lack true adaptive immunity and it solely depends on the primitive immunity called innate immunity. However, various innate immune molecules and mechanisms are identified in shrimp that plays potential role against invading bacterial, fungal and viral pathogens. Perceiving the shrimp innate immune mechanisms will contribute in developing effective vaccine strategies against major shrimp pathogens. Hence this review intends to explore the innate immune molecules of shrimp with suitable experimental evidences together with the evolution of "specific immune priming" of invertebrates. In addition, we have emphasized on the development of an effective vaccine strategy against major shrimp pathogen, white spot syndrome virus (WSSV). The baculovirus displayed rVP28 (Bac-VP28), a major envelope protein of WSSV was utilized to study its vaccine efficacy by oral route. A significant advantage of this baculovirus expression cassette is the use of WSSV-immediate early 1 (ie1) promoter that derived the abundant expression of rVP28 protein at the early stage of the infection in insect cell. The orally vaccinated shrimp with Bac-VP28 transduced successfully in the shrimp cells as well as provided highest survival rate. In support to our vaccine efficacy we analysed Pattern Recognition Proteins (PRPs) ?-1,3 glucan lipopolysaccharides (LGBP) and STAT gene profiles in the experimental shrimp. Indeed, the vaccination of shrimp with Bac-VP28 demonstrated some degree of specificity with enhanced survival rate when compared to control vaccination with Bac-wt. Hence it is presumed that the concept of "specific immune priming" in relevant to shrimp immunity is possible but may not be common to all shrimp pathogens. PMID:25083808

  15. Evolution of specific immunity in shrimp - a vaccination perspective against white spot syndrome virus.

    Science.gov (United States)

    Syed Musthaq, Syed Khader; Kwang, Jimmy

    2014-10-01

    Invertebrates lack true adaptive immunity and it solely depends on the primitive immunity called innate immunity. However, various innate immune molecules and mechanisms are identified in shrimp that plays potential role against invading bacterial, fungal and viral pathogens. Perceiving the shrimp innate immune mechanisms will contribute in developing effective vaccine strategies against major shrimp pathogens. Hence this review intends to explore the innate immune molecules of shrimp with suitable experimental evidences together with the evolution of "specific immune priming" of invertebrates. In addition, we have emphasized on the development of an effective vaccine strategy against major shrimp pathogen, white spot syndrome virus (WSSV). The baculovirus displayed rVP28 (Bac-VP28), a major envelope protein of WSSV was utilized to study its vaccine efficacy by oral route. A significant advantage of this baculovirus expression cassette is the use of WSSV-immediate early 1 (ie1) promoter that derived the abundant expression of rVP28 protein at the early stage of the infection in insect cell. The orally vaccinated shrimp with Bac-VP28 transduced successfully in the shrimp cells as well as provided highest survival rate. In support to our vaccine efficacy we analysed Pattern Recognition Proteins (PRPs) ?-1,3 glucan lipopolysaccharides (LGBP) and STAT gene profiles in the experimental shrimp. Indeed, the vaccination of shrimp with Bac-VP28 demonstrated some degree of specificity with enhanced survival rate when compared to control vaccination with Bac-wt. Hence it is presumed that the concept of "specific immune priming" in relevant to shrimp immunity is possible but may not be common to all shrimp pathogens. PMID:24780624

  16. First report on White Spot Syndrome Virus (WSSV) infection in white leg shrimp Litopenaeus vannamei (Crustacea, Penaeidae) under semi intensive culture condition in India

    OpenAIRE

    Gunalan Balakrishnan; Soundarapandian Peyail; Ramachandran Kumaran; Anand Theivasigamani; Kotiya Anil S.; Solanki Jitesh B.; Nithyamary Srinivasan

    2011-01-01

    Scientific shrimp culture began in India in the late eighties along the east coast particularly inAndrapradesh and Tamilnadu. Continuous success of shrimp culture was affected by mass mortalities ofcultured shrimp in 1994. Thereafter disease infection on survival and production of shrimps get itsimportance in culture. The present study is the first report on WSSV (white spot syndrome virus)infection in cultured Litopenaeus vannamei (Boone, 1931) in India. WSSV infection was observed on 70thda...

  17. Sensitivitas Berbagai Stadia Kepiting Bakau (Scylla paramamosain Estampador terhadap White Spot Syndrome Virus

    Directory of Open Access Journals (Sweden)

    ZAFRAN

    2005-05-01

    Full Text Available Mud crab is the most commercial crabs which are highly sold because oftheir delicious taste and high protein content. In nature, the capture intensity of this crab is very high so the population would decrease rapidly. For this reason people start to breed it. At the moment breeding of mud crab still have a lot of problems. One of those is the lower survival rate of larva stage due to the diseases. The objective of this research was to observe the sensitivity of mud crab from various stages to White Spot Syndrome Virus (WSSV. The research was conducted in Pathology laboratory, Balai Besar Riset Perikanan Budidaya Laut Gondol, Bali, January to April 2004. This experiment was run using three stages they were zoea-1, zoea-3 and crablet stage. The treatment of this research were: for zoea-1 and zoea-3 stage, i.e. treatment (A the larvae were bathed for 2 hours in stock WSSV suspension, in which the solution was diluted for 150 times; treatment (B the larvae were bathed for 2 hours in stock WSSV suspension, in which the solution was diluted for 15000 times; C as control, without WSSV. For crablet stage the WSSV infection was done by feeding them with tiger shrimp which infected by WSSV. ANOVA used to analyze the different effect of infection treatment, i.e. diluted 150x, 15000x, and control to survival rate, then continue DMRT test in 5% level. T test used to compare two treatments i.e. infection treatment with feed and without infection. PCR method used to detect WSSV of infection in zoea-1, zoea-3 and crablet stages. The results showed that for zoea-1 and zoea-3 stage, increasing concentration virus showed significant difference respect, showed by decreasing of survival rate. In crablet stage the WSSV infection by feeding can decrease survival rate. In proving the dead of mud crab zoea-1, zoea-3 and crablet were due to the infection of WSSV, the PCR method was used. The result showed that look PCR product was revenged in the cell with size of band were as expractise. According to this result can be concluded that zoea-1, zoea-3, and crablet stages of tested mud crab were sensitive to WSSV infection.

  18. Identification and characterization of a new E3 ubiquitin ligase in white spot syndrome virus involved in virus latency

    Directory of Open Access Journals (Sweden)

    Kwang Jimmy

    2008-12-01

    Full Text Available Abstract White spot syndrome virus (WSSV is one major pathogen in shrimp aquaculture. WSSV ORF403 is predicted to encode a protein of 641 amino acids, which contains a C3H2C2 RING structure. In the presence of an E2 conjugating enzyme from shrimp, WSSV403 can ubiquitinate itself in vitro, indicating it can function as a viral E3 ligase. Besides, WSSV403 E3 ligase can be activated by a series of E2 variants. Based on RT-PCR and Real time PCR, we detected transcription of WSSV403 in the commercial specific-pathogen-free (SPF shrimp, suggesting its role as a latency-associated gene. Identified in yeast two-hybrid screening and verified by pull-down assays, WSSV403 is able to bind to a shrimp protein phosphatase (PPs, which was characterized before as an interaction partner for another latent protein WSSV427. Our studies suggest that WSSV403 is a regulator of latency state of WSSV by virtue of its E3 ligase function.

  19. Validation of a Commercial Insulated Isothermal PCR-based POCKIT Test for Rapid and Easy Detection of White Spot Syndrome Virus Infection in Litopenaeus vannamei

    OpenAIRE

    Tsai, Yun-Long; Wang, Han-Ching; Lo, Chu-Fang; Tang-Nelson, Kathy; Lightner, Donald; Ou, Bor-Rung; Hour, Ai-Ling; Tsai, Chuan-Fu; Yen, Cheng-Chi; Chang, Hsiao-Fen Grace; Teng, Ping-Hua; Lee, Pei-Yu

    2014-01-01

    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, ...

  20. PmRab7 Is a VP28-Binding Protein Involved in White Spot Syndrome Virus Infection in Shrimp

    OpenAIRE

    Sritunyalucksana, Kallaya; Wannapapho, Wanphen; Lo, Chu Fang; Flegel, Timothy W

    2006-01-01

    Our aim was to isolate and characterize white spot syndrome virus (WSSV)-binding proteins from shrimp. After a blot of shrimp hemocyte membrane proteins was overlaid with a recombinant WSSV envelope protein (rVP28), the reactive bands on the blot were detected using anti-VP28 antibody. Among three membrane-associated molecules identified by liquid chromatography-tandem mass spectrometry, there was a 25-kDa protein that bound to both rVP28 and WSSV. Since it had a primary structure with high h...

  1. Production of polyclonal antiserum specific to the 27.5 kDa envelope protein of white spot syndrome virus

    OpenAIRE

    You, Z.O.; Nadala, E.C.B.; J. S. Yang; Hulten, M.C.W., van; Loh, P.C.

    2002-01-01

    A truncated version of the white spot syndrome virus (WSSV) 27.5 kDa envelope protein was expressed as a histidine tag fusion protein in Escherichia coli. The bacterial expression system allowed the production of up to 10 mg of purified recombinant protein per liter of bacterial culture. Antiserum from a rabbit immunized with the recombinant protein recognized the 27.5 kDa viral envelope protein of WSSV isolated from different geographic regions. The antiserum did not recognize any of the oth...

  2. A gC1qR Prevents White Spot Syndrome Virus Replication in the Freshwater Crayfish Pacifastacus leniusculus ?

    OpenAIRE

    Watthanasurorot, Apiruck; Jiravanichpaisal, Pikul; Söderhäll, Irene; Söderhäll, Kenneth

    2010-01-01

    The gC1qR/p32 protein is a multiple receptor for several proteins and pathogens. We cloned a gC1qR homologue in a crustacean, Pacifastacus leniusculus, and analyzed the expression of P. leniusculus C1qR (PlgC1qR) in various tissues. The gC1qR/p32 transcript was significantly enhanced by white spot syndrome virus (WSSV) infection 6 h after viral infection both in vitro in a hematopoietic tissue cell culture (Hpt) and in vivo compared to appropriate controls. Moreover, PlgC1qR silencing in both...

  3. A Novel C-Type Lectin from the Shrimp Litopenaeus vannamei Possesses Anti-White Spot Syndrome Virus Activity?

    OpenAIRE

    Zhao, Zhi-Ying; Yin, Zhi-Xin; Xu, Xiao-Peng; Weng, Shao-ping; Rao, Xia-Yu; Dai, Zong-Xian; Luo, Yong-wen; Yang, Gan; Li, Zong-Sheng; Guan, Hao-Ji; Li, Se-Dong; Chan, Siu-Ming; Yu, Xiao-Qiang; He, Jian-guo

    2008-01-01

    C-type lectins play key roles in pathogen recognition, innate immunity, and cell-cell interactions. Here, we report a new C-type lectin (C-type lectin 1) from the shrimp Litopenaeus vannamei (LvCTL1), which has activity against the white spot syndrome virus (WSSV). LvCTL1 is a 156-residue polypeptide containing a C-type carbohydrate recognition domain with an EPN (Glu99-Pro100-Asn101) motif that has a predicted ligand binding specificity for mannose. Reverse transcription-PCR analysis reveale...

  4. The Role of Aldehyde Dehydrogenase and Hsp70 in Suppression of White Spot Syndrome Virus Replication at High Temperature?

    OpenAIRE

    Lin, Ying-Ru; Hung, Hsiao-Chun; Leu, Jiann-Horng; Wang, Hao-Ching; Kou, Guang-Hsiung; Lo, Chu-Fang

    2011-01-01

    High temperature (32 to 33°C) has been shown to reduce mortality in white spot syndrome virus (WSSV)-infected shrimps, but the mechanism still remains unclear. Here we show that in WSSV-infected shrimps cultured at 32°C, transcriptional levels of representative immediate-early, early, and late genes were initially higher than those at 25°C. However, neither the IE1 nor VP28 protein was detected at 32°C, suggesting that high temperature might inhibit WSSV protein synthesis. Two-dimensional gel...

  5. White Spot Syndrome Virus Annexes a Shrimp STAT To Enhance Expression of the Immediate-Early Gene ie1?

    OpenAIRE

    Liu, Wang-Jing; Chang, Yun-Shiang; Wang, Andrew H.-J.; Kou, Guang-Hsiung; Lo, Chu-Fang

    2006-01-01

    Although the Janus kinase-signal transducer and activator of transcription (JAK-STAT) signaling pathway is part of the antiviral response in arthropods such as Drosophila, here we show that white spot syndrome virus (WSSV) uses a shrimp STAT as a transcription factor to enhance viral gene expression in host cells. In a series of deletion and mutation assays using the WSSV immediate-early gene ie1 promoter, which is active in shrimp cells and also in insect Sf9 cells, an element containing a S...

  6. Collaboration between a soluble C-type lectin and calreticulin facilitates white spot syndrome virus infection in shrimp.

    Science.gov (United States)

    Wang, Xian-Wei; Xu, Yi-Hui; Xu, Ji-Dong; Zhao, Xiao-Fan; Wang, Jin-Xing

    2014-09-01

    White spot syndrome virus (WSSV) mainly infects crustaceans through the digestive tract. Whether C-type lectins (CLs), which are important receptors for many viruses, participate in WSSV infection in the shrimp stomach remains unknown. In this study, we orally infected kuruma shrimp Marsupenaeus japonicus to model the natural transmission of WSSV and identified a CL (designated as M. japonicus stomach virus-associated CL [MjsvCL]) that was significantly induced by virus infection in the stomach. Knockdown of MjsvCL expression by RNA interference suppressed the virus replication, whereas exogenous MjsvCL enhanced it. Further analysis by GST pull-down and coimmunoprecipitation showed that MjsvCL could bind to viral protein 28, the most abundant and functionally relevant envelope protein of WSSV. Furthermore, cell-surface calreticulin was identified as a receptor of MjsvCL, and the interaction between these proteins was a determinant for the viral infection-promoting activity of MjsvCL. The MjsvCL-calreticulin pathway facilitated virus entry likely in a cholesterol-dependent manner. This study provides insights into a mechanism by which soluble CLs capture and present virions to the cell-surface receptor to facilitate viral infection. PMID:25070855

  7. A pilot metabolic profiling study in hepatopancreas of Litopenaeus vannamei with white spot syndrome virus based on ¹H NMR spectroscopy.

    Science.gov (United States)

    Liu, Peng-fei; Liu, Qing-hui; Wu, Yin; Jie, Huang

    2015-01-01

    White spot syndrome virus, which was a pathogen first found in 1992, had emerged globally affecting shrimp populations in aquaculture. Here, we comprehensively analyzed the metabolic changes of hepatopancreas from Litopenaeus vannamei which were infected with white spot syndrome virus by (1)H nuclear magnetic resonance (NMR). Through the NOESYPR1D spectrum combined with multi-variate pattern recognition analysis, including principal component analysis (PCA) and orthogonal partial least-squares discriminant analysis (OPLS-DA) models, significantly metabolic changes were observed in WSSV-infected groups compared with the control groups. In the first 48 h, ?-glucose and ?-glucose were higher in the WSSV-infected group. Meanwhile, acetate, lactate, N-acetyl glycoprotein signals, lysine, tyrosine and lipid were significantly decreased in the WSSV-infected group. These results suggest that WSSV caused absorption inhibition of amino acids and disturbed protein metabolism as well as cell metabolism in favor of its replication. Our findings could also contribute to further understanding of disease mechanisms. PMID:25450952

  8. Sequencing and De Novo Analysis of the Hemocytes Transcriptome in Litopenaeus vannamei Response to White Spot Syndrome Virus Infection

    Science.gov (United States)

    Xue, Shuxia; Liu, Yichen; Zhang, Yichen; Sun, Yan; Geng, Xuyun; Sun, Jinsheng

    2013-01-01

    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. PMID:24204661

  9. 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. PMID:25189688

  10. Identification and characterization of a prawn white spot syndrome virus gene that encodes an envelope protein VP31

    International Nuclear Information System (INIS)

    Based on a combination of SDS-PAGE and mass spectrometry, a protein with an apparent molecular mass of 31 kDa (termed as VP31) was identified from purified shrimp white spot syndrome virus (WSSV) envelope fraction. The resulting amino acid (aa) sequence matched an open reading frame (WSV340) of the WSSV genome. This ORF contained 783 nucleotides (nt), encoding 261 aa. A fragment of WSV340 was expressed in Escherichia coli as a glutathione S-transferase (GST) fusion protein with a 6His-tag, and then specific antibody was raised. Western blot analysis and the immunoelectron microscope method (IEM) confirmed that VP31 was present exclusively in the viral envelope fraction. The neutralization experiment suggested that VP31 might play an important role in WSSV infectivity

  11. Identification of the interaction domains of white spot syndrome virus envelope proteins VP28 and VP24.

    Science.gov (United States)

    Li, Zaipeng; Chen, Weiyu; Xu, Limei; Li, Fang; Yang, Feng

    2015-03-16

    VP28 and VP24 are two major envelope proteins of white spot syndrome virus (WSSV). The direct interaction between VP28 and VP24 has been described in previous studies. In this study, we confirmed this interaction and mapped the interaction domains of VP28 and VP24 by constructing a series of deletion mutants. By co-immunoprecipitation, two VP28-binding domains of VP24 were located at amino acid residues 46-61 and 148-160, while VP24-binding domain of VP28 was located at amino acid residues 31-45. These binding domains were further corroborated by peptide blocking assay, in which synthetic peptides spanning the binding domains were able to inhibit VP28-VP24 interaction, whereas same-size control peptides from non-binging regions did not. PMID:25637460

  12. Localization of VP28 on the baculovirus envelope and its immunogenicity against white spot syndrome virus in Penaeus monodon

    International Nuclear Information System (INIS)

    White spot syndrome virus (WSSV) is a large dsDNA virus responsible for white spot disease in shrimp and other crustaceans. VP28 is one of the major envelope proteins of WSSV and plays a crucial role in viral infection. In an effort to develop a vaccine against WSSV, we have constructed a recombinant baculovirus with an immediate early promoter 1 which expresses VP28 at an early stage of infection in insect cells. Baculovirus expressed rVP28 was able to maintain its structural and antigenic conformity as indicated by immunofluorescence assay and western blot analysis. Interestingly, our results with confocal microscopy revealed that rVP28 was able to localize on the plasma membrane of insect cells infected with recombinant baculovirus. In addition, we demonstrated with transmission electron microscopy that baculovirus successfully acquired rVP28 from the insect cell membrane via the budding process. Using this baculovirus displaying VP28 as a vaccine against WSSV, we observed a significantly higher survival rate of 86.3% and 73.5% of WSSV-infected shrimp at 3 and 15 days post vaccination respectively. Quantitative real-time PCR also indicated that the WSSV viral load in vaccinated shrimp was significantly reduced at 7 days post challenge. Furthermore, our RT-PCR and immunohistochemistry results demonstrated that the recombinant baculovirus was able to express VP28 in vivo in shrimp tissues. This study will be of considerable significance in elucidating the morphogenesignificance in elucidating the morphogenesis of WSSV and will pave the way for new generation vaccines against WSSV.

  13. Inactivation of White Spot Syndrome Virus (WSSV) by normal rabbit serum: Implications for the role of the envelope protein VP28 in WSSV infection of shrimp

    OpenAIRE

    Robalino, J.; PAYNE, C; Parnell, P; Shepard, E; Grimes, A.C.; Metz, A.; Prior, S.; Witteveldt, J.; J. M. Vlak; P. S. Gross; Warr, G.; Browdy, C.L.

    2006-01-01

    White Spot Syndrome Virus (WSSV) is a highly pathogenic and prevalent virus affecting crustacea. A number of WSSV envelope proteins, including vp28, have been proposed to be involved in viral infectivity based on the ability of specific antibodies to attenuate WSSV-induced mortality in vivo. In the present study, a series of monoclonal and polyclonal antibodies targeting vp28 were tested for their ability to neutralize WSSV infectivity, with the purpose of identifying epitopes potentially inv...

  14. The endemic region and infection regimes of the White Spot Syndrome virus (WSSV in shrimp farms in northwestern México

    Directory of Open Access Journals (Sweden)

    Héctor Manuel Esparza Leal

    2012-09-01

    Full Text Available Shrimp farming with a value annually of US$711 million approximately, is one of the most important primary activities in Mexico. However, shrimp farming has had to face various problems that have limited their development, within which the mortality caused by the white spot syndrome virus (WSSV is the most important. To have scientific elements to focus on preventive health management actions is necessary to know, among other factors, aspects of the epidemiologyof white spot disease (WSD. Therefore this study focused on delimiting the endemic region for WSD and its temporal regimes of infection and discusses possible risk factors related to outbreaks of the disease in shrimp farms of northwestern Mexico. We analyzed information from the databases of the State Committees of Aquaculture Health of Baja California Sur, Sonora, Sinaloa and Nayarit, as well as data of Integrated Program on Shrimp Aquaculture Health (PISA 2007-2008 and the Strategic Alliance Network Aquaculture Industry Innovation (AERI-2008. Data analysis showed that, for the shrimp production cycles of 2007-2008, white spot syndrome virus (WSSV was endemic to the region of Tuxpan, Nayarit in the south and to Agiabampo, Sonora in the north. Spring outbreaks of WSD in the fishfarms had a spatiotemporal distribution, indicating three infections regimes: (1 March-April in the southern shrimpfarming region (Local Aquaculture Health Boards [LAHBs] of Mazatlan, El Rosario, Escuinapa, Tecuala, and Tuxpan; 2 April-May in the central region (LAHBs of Navolato Norte, Navolato Sur, and El dorado; and (3 May-June in the northern region (LAHBs of Agiabampo-Sonora, Ahome, Guasave Norte and Sur. The WSD were consistent between 2007 and 2008, with slight variations among some LAHBs, with respect to the onset or presence of spring WSD outbreaks. It shows the association of infection regimes throughout the region endemic with the location of Mazatlan,Pescadero and Farallon oceanographic basins according to the increasing differential temperature within them, which may be a determinant factor for the presence of WSD outbreaks.

  15. White spot syndrome virus (WSSV) genome stability maintained over six passages through three different penaeid shrimp species.

    Science.gov (United States)

    Sindhupriya, M; Saravanan, P; Otta, S K; Amarnath, C Bala; Arulraj, R; Bhuvaneswari, T; Praveena, P Ezhil; Jithendran, K P; Ponniah, A G

    2014-08-21

    White spot syndrome virus (WSSV) replicates rapidly, can be extremely pathogenic and is a common cause of mass mortality in cultured shrimp. Variable number tandem repeat (VNTR) sequences present in the open reading frame (ORF)94, ORF125 and ORF75 regions of the WSSV genome have been used widely as genetic markers in epidemiological studies. However, reports that VNTRs might evolve rapidly following even a single transmission through penaeid shrimp or other crustacean hosts have created confusion as to how VNTR data is interpreted. To examine VNTR stability again, 2 WSSV strains (PmTN4RU and LvAP11RU) with differing ORF94 tandem repeat numbers and slight differences in apparent virulence were passaged sequentially 6 times through black tiger shrimp Penaeus monodon, Indian white shrimp Feneropenaeus indicus or Pacific white leg shrimp Litopenaeus vannamei. PCR analyses to genotype the ORF94, ORF125 and ORF75 VNTRs did not identify any differences from either of the 2 parental WSSV strains after multiple passages through any of the shrimp species. These data were confirmed by sequence analysis and indicate that the stability of the genome regions containing these VNTRs is quite high at least for the WSSV strains, hosts and number of passages examined and that the VNTR sequences thus represent useful genetic markers for studying WSSV epidemiology. PMID:25144114

  16. The first detection of white spot syndrome virus in naturally infected cultured Chinese mitten crabs, Eriocheir sinensis in China.

    Science.gov (United States)

    Ding, Zhengfeng; Yao, Yufeng; Zhang, Fengxiang; Wan, Jinjuan; Sun, Mengling; Liu, Hongyan; Zhou, Gang; Tang, Jianqing; Pan, Jianlin; Xue, Hui; Zhao, Ziming

    2015-08-01

    An epidemic with a high mortality rate (80-100%) recently occurred in the cultured Chinese mitten crab, Eriocheir sinensis, which is a very important economic crustacean species in China. Using negative stain, histopathology and nested PCR supplemented by sequencing we identified white spot syndrome virus (WSSV) in these crabs. Challenge experiments revealed that the disease was caused by WSSV and confirmed the crab's susceptibility to this virus, which was consistent with previous laboratory-based studies. A cumulative mortality of 100% was observed within 10 days post WSSV injection. This is the first report of WSSV-associated disease outbreaks in the Chinese mitten crab, which is normally reported as an important penaeid-shrimp viral pathogen. Furthermore, this is only the second report to describe a significant pathogen in pond-cultured E. sinensis. These results will enhance the early diagnosis of WSSV in the crab farms and help in monitoring efforts directed at determining the prevalence of the virus in E. sinensis. PMID:25907468

  17. Antilipopolysaccharide factor interferes with white spot syndrome virus replication in vitro and in vivo in the crayfish Pacifastacus leniusculus.

    Science.gov (United States)

    Liu, Haipeng; Jiravanichpaisal, Pikul; Söderhäll, Irene; Cerenius, Lage; Söderhäll, Kenneth

    2006-11-01

    In a study of genes expressed differentially in the freshwater crayfish Pacifastacus leniusculus infected experimentally with the white spot syndrome virus (WSSV), one protein, known as antilipopolysaccharide factor (ALF), was chosen, among those whose transcript levels increased upon viral infection, for further studies. ALF RNA interference (RNAi) experiments in whole animals and in cell cultures indicated that ALF can protect against WSSV infection, since knockdown of ALF by RNAi specifically resulted in higher rates of viral propagation. In a cell culture of hematopoietic tissue (Hpt) from P. leniusculus, quantitative PCR showed that knockdown of ALF by RNAi resulted into WSSV levels that were about 10-fold higher than those treated with control double-stranded RNA (dsRNA). In addition, RNAi experiments with other crayfish genes that had been found to be up-regulated by a WSSV infection did not result in any changes of viral loads. Thus, the cell culture does not respond to dsRNA in a similar manner, as shown earlier for dsRNA injected into shrimp, which gave a higher degree of resistance to WSSV infection. If ALF transcription in whole animals was stimulated by the administration of UV-treated WSSV, a partial protection against a subsequent challenge with the active virus was conferred to the host. This is the first crustacean gene product identified with the capacity to interfere with replication of this important pathogen. PMID:17041217

  18. An immediate-early protein of white spot syndrome virus modulates the phosphorylation of focal adhesion kinase of shrimp.

    Science.gov (United States)

    Lu, Huasong; Ruan, Lingwei; Xu, Xun

    2011-10-25

    WSSV interacts with integrin during infection of shrimps and modulate the focal adhesion kinase which is known as a regulator of several downstream signaling pathways. Viral protein kinases are thought to be important for virus infection by regulating the host signaling pathways. WSV083 is an immediate-early gene of white spot syndrome virus that contains a Ser/Thr protein kinase domain. So, does WSSV modulate FAK phosphorylation via the WSV083 molecule? In this study, co-transfection of WSV083 and MjFAK genes proceeded in insect cells revealed that the MjFAK phosphorylation and cell adhesion activity could be inhibited by the expression of WSV083. Kinase domain mutants of WSV083 lost its ability of inhibiting FAK phosphorylation. Moreover, silencing of FAK gene through RNAi accelerated the shrimp death rate upon WSSV challenge. These results demonstrate for the first time that modulation of FAK phosphorylation by WSV083 plays a critical role in the pathogenesis of WSSV infection. PMID:21908012

  19. Microarray and RT-PCR screening for white spot syndrome virus immediate-early genes in cycloheximide-treated shrimp

    International Nuclear Information System (INIS)

    Here, we report for the first time the successful use of cycloheximide (CHX) as an inhibitor to block de novo viral protein synthesis during WSSV (white spot syndrome virus) infection. Sixty candidate IE (immediate-early) genes were identified using a global analysis microarray technique. RT-PCR showed that the genes corresponding to ORF126, ORF242 and ORF418 in the Taiwan isolate were consistently CHX-insensitive, and these genes were designated ie1, ie2 and ie3, respectively. The sequences for these IE genes also appear in the two other WSSV isolates that have been sequenced. Three corresponding ORFs were identified in the China WSSV isolate, but only an ORF corresponding to ie1 was predicted in the Thailand isolate. In a promoter activity assay in Sf9 insect cells using EGFP (enhanced green fluorescence protein) as a reporter, ie1 showed very strong promoter activity, producing higher EGFP signals than the insect Orgyia pseudotsugata multicapsid nuclear polyhedrosis virus (OpMNPV) ie2 promoter

  20. A sensitive and specific hyperbranched rolling circle amplification assay and test strip for white spot syndrome virus.

    Science.gov (United States)

    Zhao, Yu-Ran; Yin, Wei-Li; Yue, Zhi-Qin; Li, Ba-Fang

    2014-01-01

    White spot syndrome virus (WSSV) is a global threat to the prawn industry, and there is no simple method for field-based testing of this virus. We designed a padlock probe and primers to the capsid protein gene VP28 of WSSV, and established a hyperbranched rolling circle amplification (HRCA) assay and a corresponding strip-based test. The assay and the test strip both had similar high accuracy and specificity, and their sensitivity was about 10 copies/?L, which is 100 times higher than conventional PCR. In this study, 68 batches of prawns were tested for WSSV with the HRCA assay and test strip, and the results were compared with the PCR assay. The results indicated that both the assay and test strip had accuracy similar to each other and to the PCR results. However, the assay and strip were more sensitive and user-friendly than PCR. Establishment of this method will provide a rapid detection of WSSV and also a basis for field-based detection of animal disease. PMID:25902992

  1. Molecular characterization of LvAV in response to white spot syndrome virus infection in the Pacific white shrimp (Litopenaeus vannamei).

    Science.gov (United States)

    He, Shulin; Song, Lei; Qian, Zhaoying; Hou, Fujun; Liu, Yongjie; Wang, Xianzong; Peng, Zhangming; Sun, Chengbo; Liu, Xiaolin

    2015-07-01

    Litopenaeus vannamei is the most important farmed shrimp species globally, but its production is affected by several factors, including infectious disease. White spot syndrome virus (WSSV), in particular, causes significant shrimp losses. To understand the shrimp's immune response against WSSV, we cloned LvAV from L.?vannamei and analyzed its expression pattern in different tissues, in addition to its expression following infection. We employed dsRNA and recombinant (r)LvAV to explore the potential role of LvAV in shrimp immunity when infected with WSSV. We find that LvAV is a C-type Lectin composed of 176 amino acids with a signal peptide and a specific C-type Lectin-type domain (CTLD). It shares 81% amino acid similarity with PmAV, an antiviral-like C-type Lectin from Penaeus monodom, and it is highly expressed in the hepatopancreas. Its expression is affected by infection with both WSSV and V.?parahaemolyticus. Significantly, injection with rLvAV slowed WSSV replication, while injection with LvAV dsRNA initially led to enhanced virus propagation. Surprisingly, LvAV dsRNA subsequently led to a dramatic decrease in viral load in the later stages of infection, suggesting that LvAV may be subverted by WSSV to enhance viral replication or immune avoidance. Our results indicate that LvAV plays an important, but potentially complex role in the Pacific white shrimp's immune defense. PMID:25735872

  2. Identification of highly expressed host microRNAs that respond to white spot syndrome virus infection in the Pacific white shrimp Litopenaeus vannamei (Penaeidae).

    Science.gov (United States)

    Zeng, D G; Chen, X L; Xie, D X; Zhao, Y Z; Yang, Q; Wang, H; Li, Y M; Chen, X H

    2015-01-01

    MicroRNAs (miRNAs) are known to play an important role in regulating both adaptive and innate immunity. Pacific white shrimp (Litopenaeus vannamei) is the most widely farmed crustacean species in the world. However, little is known about the role miRNAs play in shrimp immunity. To understand the impact of viral infection on miRNA expression in shrimp, we used high-throughput sequencing technology to sequence two small RNA libraries prepared from L. vannamei under normal and white spot syndrome virus (WSSV) challenged conditions. Approximately 19,312,189 and 39,763,551 raw reads corresponding to 17,414,787 and 28,633,379 high-quality mappable reads were obtained from the two libraries, respectively. Twelve conserved miRNAs and one novel miRNA that were highly expressed (>100 RPM) in L. vannamei were identified. Of the identified miRNAs, 8 were differentially expressed in response to the virus infection, of which 1 was upregulated and 7 were downregulated. The prediction of miRNA targets showed that the target genes of the differentially expressed miRNAs were related to immunity, apoptosis, and development functions. Our study provides the first characterization of L. vannamei miRNAs in response to WSSV infection, which will help to reveal the roles of miRNAs in the antiviral mechanisms of shrimp. PMID:25966256

  3. Pupil - white spots

    Science.gov (United States)

    ... pupil may appear red. This is called the "red reflex" by health care providers and is normal. Sometimes, ... the eye may appear white, or the normal red reflex may appear to be white. This not a ...

  4. Transmission of white spot syndrome virus in improved-extensive and semi-intensive shrimp production systems: A molecular epidemiology study

    OpenAIRE

    Tuyet Hoa, T.T.; Zwart, M.P.; N.T. Phuong; J. M. Vlak; Jong, M.C.M., de

    2011-01-01

    Experimental evidence suggests that white spot syndrome virus (WSSV) can be transmitted horizontally through water, via carrier organisms and/or by cannibalism of infected shrimp, but also vertically through infected broodstock. However the mode(s) of WSSV transmission in shrimp farming systems and the epidemiological consequences are not well understood. In this study, molecular markers have been used to analyse the spread of WSSV within and between farms in Viet Nam for two different farmin...

  5. Identification of White Spot Syndrome Virus Latency-Related Genes in Specific-Pathogen-Free Shrimps by Use of a Microarray

    OpenAIRE

    Khadijah, Siti; Neo, Soek Ying; Hossain, M.S.; Miller, Lance D; Mathavan, S.; Kwang, Jimmy

    2003-01-01

    To investigate whether specific-pathogen-free (SPF) shrimps are asymptomatic carriers of white spot syndrome virus (WSSV), we used a WSSV-specific DNA microarray to measure WSSV gene expression in SPF and WSSV-infected shrimps. Three WSSV genes were found to be relatively highly expressed in SPF shrimps. Reverse transcription-PCR using nested primers as well as real-time detection confirmed that these genes have no detectable counterparts in GenBank; structural analysis of the putative protei...

  6. White Spot Syndrome Virus Proteins and Differentially Expressed Host Proteins Identified in Shrimp Epithelium by Shotgun Proteomics and Cleavable Isotope-Coded Affinity Tag? †

    OpenAIRE

    Wu, Jinlu; Lin, Qingsong; Lim, Teck Kwang; Liu, Tiefei; Hew, Choy-leong

    2007-01-01

    Shrimp subcuticular epithelial cells are the initial and major targets of white spot syndrome virus (WSSV) infection. Proteomic studies of WSSV-infected subcuticular epithelium of Penaeus monodon were performed through two approaches, namely, subcellular fractionation coupled with shotgun proteomics to identify viral and host proteins and a quantitative time course proteomic analysis using cleavable isotope-coded affinity tags (cICATs) to identify differentially expressed cellular proteins. P...

  7. White spot syndrome virus (WSSV) risk factors associated with farming practices in polyculture and monoculture P. monodon farms in the Philippines

    OpenAIRE

    Tendencia Alapide, E.; Bosma, R. H.; Verreth, J.A.J.

    2011-01-01

    White spot sydrome virus (WSSV) is one of the most important viral disease of shrimp. Several studies to control the disease have been done. Tank experiments identified WSSV risk factors related to the physico chemical properties of the water. A few studies reported pond level WSSV risk factors. This study identifies the risk factors associated with essentially two different farming systems: polyculture and semi-intensive monoculture of Penaeus monodon. Data were gathered from a total of 174 ...

  8. Increased tolerance of Litopenaeus vannamei to white spot syndrome virus (WSSV) infection after oral application of the viral envelope protein VP28

    OpenAIRE

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

    2006-01-01

    It has been generally accepted that invertebrates such as shrimp do not have an adaptive immune response system comparable to that of vertebrates. However, in the last few years, several studies have suggested the existence of such a response in invertebrates. In one of these studies, the shrimp Penaeus monodon showed increased protection against white spot syndrome virus (WSSV) using a recombinant VP28 envelope protein of WSSV. In an effort to further investigate whether this increased prote...

  9. Knocking down caspase-3 by RNAi reduces mortality in Pacific white shrimp Penaeus (Litopenaeus) vannamei challenged with a low dose of white-spot syndrome virus.

    Science.gov (United States)

    Rijiravanich, Anchukorn; Browdy, Craig L; Withyachumnarnkul, Boonsirm

    2008-03-01

    Apoptosis has long been observed in viral target organs of white-spot syndrome virus (WSSV)-infected shrimp and whether the phenomenon helps the shrimp to survive the infection or is a factor leading to mortality is still controversial. If the shrimp mortality is a result of triggered apoptosis, then inactivation of caspase-3, a key protein in the induction of apoptosis, should improve shrimp survival upon challenge with WSSV. To test this prediction, we identified and characterized a caspase-3 homologue (cap-3) from the Pacific white shrimp Penaeus (Litopenaeus) vannamei and used this information to silence cap-3 expression by RNA interference prior to WSSV challenge. After confirming the efficacy of cap-3 silencing, its effects on mortality at high and low doses of WSSV were evaluated. In a high-dose WSSV challenge, cap-3 silencing had no significant effect on WSSV-induced mortality, except for a delay in mean time to death. However, at a low-dose WSSV challenge, cap-3 silencing correlated with a lower level of cumulative mortality, relative to silencing of a control gene, suggesting that apoptosis may exacerbate rather than decrease mortality in WSSV-challenged shrimp. PMID:18248799

  10. Effect of multiple infections with white spot syndrome virus and Vibrio anguillarum on Pacific white shrimp Litopenaeus vannamei (L.): mortality and viral replication.

    Science.gov (United States)

    Jang, I K; Qiao, G; Kim, S-K

    2014-10-01

    Multiple infections are commonly found in practical shrimp culture and may cause more serious consequences than infections by one pathogen only. Therefore, this study was conducted to evaluate the effect of multiple infections with white spot syndrome virus (WSSV) and Vibrio anguillarum on Pacific white shrimp Litopenaeus vannamei (L.), mortality, WSSV replication in vivo and host immune response. In the WSSV single-infection group (WSSV load, 2 × 10(2) copies ?L(-1)), mean cumulative mortality was 29.2%. In the V. anguillarum single-infection group, cumulative mortality was 12.5% when shrimp were challenged by 10(5) CFU mL(-1) of bacteria. In the co- and super-infection groups, 37.5% and 50% cumulative mortalities, respectively, were observed at a lower bacterial concentration of 10(3) CFU mL(-1), suggesting that shrimp with multiple infections died earlier and more frequently than singly infected shrimp. WSSV load after injection was tracked over time by TaqMan quantitative PCR. WSSV load increased more rapidly in the multiple-infection groups than in the single-infection group. Additionally, mRNA expression of the genes encoding prophenoloxidase 1 and 2, which are closely involved in innate immunity in shrimp, was down-regulated more extensively in multiple-infection groups than in single-infection groups, as indicated by quantitative reverse-transcription PCR. PMID:24127689

  11. PCR detection of white spot syndrome virus (WSSV from farmed Pacific white shrimp (Litopenaeus vannamei in selected sites of the Philippines

    Directory of Open Access Journals (Sweden)

    Mary Beth B. Maningas

    2011-10-01

    Full Text Available Great losses caused by white spot syndrome virus (WSSV in shrimp culture have beenattributed to poor screening procedures in farms and the lack of sufficient access to specific pathogenfree brood stock. Thus, early detection of the virus is considered the best option for shrimp farmers. Thestudy, thus, assessed viral incidence in the Philippines and partially sequenced and characterized thePhilippine WSSV isolate with regards to other isolates in GenBank. Developed primers for PCR can detecttarget genes from 0.4 pg of DNA extract from shrimp samples. PCR detection revealed that 6.67 %(1/15 of market samples from Zambales are infected with WSSV. Shrimp samples from a local shop anda public market in General Santos City showed 46.67% (7/15 and 20% (3/15 WSSV-positive samplesrespectively. Shrimp sources from Capiz and Batangas, however, showed negative detection for WSV. Nosignificant difference in the number of infected samples from the sampling sites was found. Combineddetections reveal that the Philippines has a low infection rate of 14.67%. The study has partiallysequenced and characterized Philippine isolate. During the sampling period, most shrimps in GeneralSantos City were WSSV-positive by PCR detection.

  12. Low numbers of repeat units in variable number of tandem repeats (VNTR) regions of white spot syndrome virus are correlated with disease outbreaks

    OpenAIRE

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

    2012-01-01

    White spot syndrome virus (WSSV) is the most important pathogen in shrimp farming systems worldwide including the Mekong Delta, Vietnam. The genome of WSSV is characterized by the presence of two major 'indel regions' found at ORF14/15 and ORF23/24 (WSSV-Thailand) and three regions with variable number tandem repeats (VNTR) located in ORF75, ORF94 and ORF125. In the current study, we investigated whether or not the number of repeat units in the VNTRs correlates with virus outbreak status and/...

  13. Immune gene expression profile of Penaeus monodon in response to marine yeast glucan application and white spot syndrome virus challenge.

    Science.gov (United States)

    Wilson, Wilsy; Lowman, Douglas; Antony, Swapna P; Puthumana, Jayesh; Bright Singh, I S; Philip, Rosamma

    2015-04-01

    Immunostimulant potential of eight marine yeast glucans (YG) from Candida parapsilosis R20, Hortaea werneckii R23, Candida spencermartinsiae R28, Candida haemulonii R63, Candida oceani R89, Debaryomyces fabryi R100, Debaryomyces nepalensis R305 and Meyerozyma guilliermondii R340 were tested against WSSV challenge in Penaeus monodon post larvae (PL). Structural characterization of these marine yeast glucans by proton nuclear magnetic resonance (NMR) indicated structures containing (1-6)-branched (1-3)-?-D-glucan. PL were fed 0.2% glucan incorporated diet once in seven days for a period of 45 days and the animals were challenged with white spot syndrome virus (WSSV). The immunostimulatory activity of yeast glucans were assessed pre- and post-challenge WSSV by analysing the expression profile of six antimicrobial peptide (AMP) genes viz., anti-lipopolysaccharide factor (ALF), crustin-1, crustin-2, crustin-3, penaeidin-3 and penaeidin-5 and 13 immune genes viz., alpha-2-macroglobulin (?-2-M), astakine, caspase, catalase, glutathione peroxidase, glutathione-s-transferase, haemocyanin, peroxinectin, pmCathepsinC, prophenol oxidase (proPO), Rab-7, superoxide dismutase and transglutaminase. Expression of seven WSSV genes viz., DNA polymerase, endonuclease, protein kinase, immediate early gene, latency related gene, thymidine kinase and VP28 were also analysed to detect the presence and intensity of viral infection in the experimental animals post-challenge. The study revealed that yeast glucans (YG) do possess immunostimulatory activity against WSSV and also supported higher survival (40-70 %) post-challenge WSSV. Among the various glucans tested, YG23 showed maximum survival (70.27%), followed by YG20 (66.66%), YG28 (60.97%), YG89 (58.53%), YG100 (54.05%), YG63 (48.64%), YG305 (45.7%) and YG340 (43.24%). PMID:25555812

  14. Prohibitin Interacts with Envelope Proteins of White Spot Syndrome Virus and Prevents Infection in the Red Swamp Crayfish, Procambarus clarkii

    Science.gov (United States)

    Lan, Jiang-Feng; Li, Xin-Cang; Sun, Jie-Jie; Gong, Jing; Wang, Xian-Wei; Shi, Xiu-Zhen; Shi, Li-Jie; Weng, Yu-Ding; Zhao, Xiao-Fan

    2013-01-01

    Prohibitins (PHBs) are ubiquitously expressed conserved proteins in eukaryotes that are associated with apoptosis, cancer formation, aging, stress responses, cell proliferation, and immune regulation. However, the function of PHBs in crustacean immunity remains largely unknown. In the present study, we identified a PHB in Procambarus clarkii red swamp crayfish, which was designated PcPHB1. PcPHB1 was widely distributed in several tissues, and its expression was significantly upregulated by white spot syndrome virus (WSSV) challenge at the mRNA level and the protein level. These observations prompted us to investigate the role of PcPHB1 in the crayfish antiviral response. Recombinant PcPHB1 (rPcPHB1) significantly reduced the amount of WSSV in crayfish and the mortality of WSSV-infected crayfish. The quantity of WSSV in PcPHB1 knockdown crayfish was increased compared with that in the controls. The effects of RNA silencing were rescued by rPcPHB1 reinjection. We further confirmed the interaction of PcPHB1 with the WSSV envelope proteins VP28, VP26, and VP24 using pulldown and far-Western overlay assays. Finally, we observed that the colloidal gold-labeled PcPHB1 was located on the outer surface of the WSSV, which suggests that PcPHB1 specifically binds to the envelope proteins of WSSV. VP28, VP26, and VP24 are structural envelope proteins and are essential for attachment and entry into crayfish cells. Therefore, PcPHB1 exerts its anti-WSSV effect by binding to VP28, VP26, and VP24, preventing viral infection. This study is the first report on the antiviral function of PHB in the innate immune system of crustaceans. PMID:24049173

  15. Transcriptome analysis of hemocytes and hepatopancreas in red swamp crayfish, Procambarus clarkii, challenged with white spot syndrome virus

    Directory of Open Access Journals (Sweden)

    X-Z Shi

    2010-04-01

    Full Text Available Red swamp crayfish Procambarus clarkii is used for the innate immune defense of crustaceans due to its convenience for laboratory culture and study. To know more about the transcriptome of the crayfish, we constructed and sequenced a cDNA library from a mixture of hemocytes and hepatopancreas from white spot syndrome virus (WSSV-infected crayfish. By random sequencing, we obtained 9115 high-quality expressed sequence tags with a mean length of 370 bp, representing 3033 unigenes. Most of the unigenes are first reports for the red swamp crayfish. Besides the metabolic genes, many genes that may be involved in the innate immune system of the crayfish are also obtained from the library, such as antimicrobial peptides, pattern recognition receptors, proteases and protease inhibitors, signal transduction proteins, apoptosis-, antioxidant-, and RNA interference-related proteins. We chose ten immune-related genes to analyze their expression pattern by quantitative real time polymerase chain reaction (qRT-PCR from the hemocytes of normal and WSSV-challenged crayfish. Seven of them, including anti-lipopolysaccharide factor, astacidin, crustin 1, H3 histone family 3A, serine/threonine protein kinase, TGF beta-inducible nuclear protein, and tar RNA binding protein, were upregulated after WSSV injection, but the mRNA expression levels of crustin 2, a lectin, and a digestive cysteine protease decreased after WSSV infection. Our results showed that the transcriptome analysis provides a useful resource for identification of immune related genes and understanding the immune responses of the crayfish.

  16. Molecular docking analyses of Avicennia marinaderived phytochemicals against white spot syndrome virus (WSSV) envelope protein-VP28

    OpenAIRE

    Sahu, Sunil Kumar; Kathiresan, Kandasamy; Singh, Reena; Senthilraja, Poomalai

    2012-01-01

    White spot syndrome (WSS) is one of the most common and most disastrous diseases of shrimp worldwide. It causes up to 100% mortality within 3 to 4 days in commercial shrimp farms, resulting in large economic losses to the shrimp farming industry. VP28 envelope protein of WSSV is reported to play a key role in the systemic infection in shrimps. Considering the most sombre issue of viral disease in cultivated shrimp, the present study was undertaken to substantiate the inhibition poten...

  17. Crowding of white shrimp Litopenaeus vananmei depresses their immunity to and resistance against Vibrio alginolyticus and white spot syndrome virus.

    Science.gov (United States)

    Lin, Yong-Chin; Chen, Jiann-Chu; Chen, Yu-Yuan; Yeh, Su-Tuen; Chen, Li-Li; Huang, Chien-Lun; Hsieh, Jen-Fang; Li, Chang-Che

    2015-07-01

    Immunity parameters and the expression levels of several immune-related proteins, including lipopolysaccharide and ?-glucan binding protein (LGBP), peroxinectin (PX), intergin ? (IB), prophenoloxidase (proPO) I, proPO II, ?2-macroglobulin (?2-M), cytosolic mangangese superoxide dismutase (cytMnSOD), mitochondria manganese superoxide dismutase (mtMnSOD), catalase, glutathione peroxidase (GPx), lysozyme, and penaeidin 3a were examined in white shrimp Litopenaeus vannamei reared at stocking densities of 2, 10, 20, 30, and 40 shrimp L(-1) after 3, 6, and 12 h. All immune parameters including haemocyte count, phenoloxidase (PO) activity, respiratory burst (RB), superoxide dismutase (SOD) activity, lysozyme activity, and haemolymph protein were negatively related to density and time. The PO activity, SOD activity, and lysozyme activity of shrimp reared at 10 shrimp L(-1) after 12 h significantly decreased. The transcript levels of these immune-related proteins were down-regulated in shrimp reared at 20, 30, and 40 shrimp L(-1) after 12 h. Phagocytic activity and clearance efficiency to Vibrio alginolyticus were significantly lower in shrimp reared at 30 and 40 shrimp L(-1) after 12 h. The mortality rates of shrimp reared at 20 and 40 shrimp L(-1) were significantly higher than shrimp reared at 2 shrimp L(-1) over 12-144 h and 12-48 h, respectively. Shrimp reared at high densities (>10 shrimp L(-1)) exhibited decreased resistance against pathogens as evidenced by reductions in immune parameters together with decreased expression levels of immune-related proteins, indicating perturbations of the immune system. PMID:25700787

  18. RING-H2 Protein WSSV249 from White Spot Syndrome Virus Sequesters a Shrimp Ubiquitin-Conjugating Enzyme, PvUbc, for Viral Pathogenesis

    OpenAIRE

    Wang, Zhilong; Chua, Hui Kheng; Gusti, Ayu A. R. A.; He, Fang; Fenner, Beau; Manopo, Ivanus; Wang, Hai; Kwang, Jimmy

    2005-01-01

    Modification of proteins by ubiquitin is essential for numerous cellular processes. The RING-H2 finger motif has been implicated in ubiquitin-conjugating enzyme (E2)-dependent ubiquitination. Four proteins, WSSV199, WSSV222, WSSV249, and WSSV403, from white spot syndrome virus (WSSV) contain the RING-H2 motif. Here we report that WSSV249 physically interacts with a shrimp ubiquitin-conjugating enzyme, PvUbc, and mediates ubiquitination through its RING-H2 motif in the presence of E1 and PvUbc...

  19. Crystal Structures of Major Envelope Proteins VP26 and VP28 from White Spot Syndrome Virus Shed Light on Their Evolutionary Relationship?

    OpenAIRE

    Tang, Xuhua; Wu, Jinlu; Sivaraman, J.; Hew, Choy Leong

    2007-01-01

    White spot syndrome virus (WSSV) is a virulent pathogen known to infect various crustaceans. It has bacilliform morphology with a tail-like appendage at one end. The envelope consists of four major proteins. Envelope structural proteins play a crucial role in viral infection and are believed to be the first molecules to interact with the host. Here, we report the localization and crystal structure of major envelope proteins VP26 and VP28 from WSSV at resolutions of 2.2 and 2.0 Å, respectively...

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

    OpenAIRE

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

    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 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...

  1. PmVRP15, a Novel Viral Responsive Protein from the Black Tiger Shrimp, Penaeus monodon, Promoted White Spot Syndrome Virus Replication

    OpenAIRE

    Vatanavicharn, Tipachai; Prapavorarat, Adisak; Jaree, Phattarunda; Somboonwiwat, Kunlaya; Tassanakajon, Anchalee

    2014-01-01

    Suppression subtractive hybridization of Penaeus monodon hemocytes challenged with white spot syndrome virus (WSSV) has identified the viral responsive gene, PmVRP15, as the highest up-regulated gene ever reported in shrimps. Expression analysis by quantitative real time RT-PCR revealed 9410–fold up-regulated level at 48 h post WSSV injection. Tissue distribution analysis showed that PmVRP15 transcript was mainly expressed in the hemocytes of shrimp. The full-length cDNA of PmVRP15 transcript...

  2. Penaeus monodon TATA Box-Binding Protein Interacts with the White Spot Syndrome Virus Transactivator IE1 and Promotes Its Transcriptional Activity?

    OpenAIRE

    Liu, Wang-Jing; Chang, Yun-Shiang; Huang, Wei-Tung; Chen, I-Tung; Wang, K. C. Han-Ching; Kou, Guang-Hsiung; Lo, Chu-Fang

    2011-01-01

    We show here that the white spot syndrome virus (WSSV) immediate-early protein IE1 interacts with the Penaeus monodon TATA box-binding protein (PmTBP) and that this protein-protein interaction occurs in the absence of any other viral or cellular proteins or nucleic acids, both in vitro and in vivo. Mapping studies using enhanced green fluorescent protein (EGFP) fusion proteins containing truncations of IE1 and PmTBP delimited the interacting regions to amino acids (aa) 81 to 180 in IE1 and, e...

  3. VP26 of White Spot Syndrome Virus Functions as a Linker Protein between the Envelope and Nucleocapsid of Virions by Binding with VP51?

    OpenAIRE

    Wan, Qiang; Xu, Limei; Feng YANG

    2008-01-01

    The envelopment of the nucleocapsid is an important step in white spot syndrome virus (WSSV) assembly. Previous studies showed that VP26, a major envelope protein of WSSV, can interact with viral nucleocapsid. In this study, using the biotin label transfer technique, we found that the biotin label was transferred from Bio-rVP26 to the viral capsid protein VP51 or from Bio-MBP-VP51 to VP26. Far-Western analyses provided further evidence for direct interaction between VP26 and VP51. Therefore, ...

  4. White Spot Syndrome Virus Open Reading Frame 222 Encodes a Viral E3 Ligase and Mediates Degradation of a Host Tumor Suppressor via Ubiquitination

    OpenAIRE

    He, Fang; Fenner, Beau J.; Godwin, Andrew K; Kwang, Jimmy

    2006-01-01

    We have characterized a white spot syndrome virus (WSSV) RING-H2-type protein, WSSV222, which is involved in ubiquitination. WSSV222 exhibits RING-H2-dependent E3 ligase activity in vitro in the presence of the specific conjugating enzyme UbcH6. Mutations in the RING-H2 domain abolished WSSV222-dependent ubiquitination, revealing the importance of this domain in WSSV222 function. Yeast two-hybrid and pull-down analyses revealed that WSSV222 interacts with a shrimp tumor suppressor-like protei...

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

    Science.gov (United States)

    Ananphongmanee, Vorawit; Srisala, Jiraporn; Sritunyalucksana, Kallaya; Boonchird, Chuenchit

    2015-01-01

    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 ?-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 WSSV by oral administration. PMID:26083446

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

    Science.gov (United States)

    Ananphongmanee, Vorawit; Srisala, Jiraporn; Sritunyalucksana, Kallaya; Boonchird, Chuenchit

    2015-01-01

    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 ?-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 WSSV by oral administration. PMID:26083446

  7. [Comparative analysis of variable region of white spot syndrome virus genome in Penaeus vannamei in Guangxi, China].

    Science.gov (United States)

    Tong, Gui-Xiang; Li, Xiao-Zheng; Wei, Xin-Xian; Ye, Xin-Yu; Wu, Ming-Yuan; Qin, Zhen-Fa; Lan, Liu-Chun; Zhou, Jing-Jing

    2014-01-01

    Comparative analysis of variable region ORF14/15 genes of white spot syndrome virus (WSSV) genome in Guangxi Penaeus vannamei (P. vannamei) could provide useful information for the evaluation of genetic diversity and genetic evolutionary relationship among WSSV isolates from Guangxi, China and other places. Based on geographical and temporal considerations, 40 WSSV-positive P. vannamei samples were collected during the period between May 2010 and July 2013 from Beihai, Qinzhou, and Fangchenggang, which were the main P. vannamei production areas in Guangxi, and the variable region ORF14/15 genes of the WSSV genome from all infected samples were amplified by PCR and then subjected to cloning and sequence analysis. Pairwise and multiple alignment analysis was then conducted to evaluate the degree of genetic divergence between different strains. The variable region ORF14/15 genes from 25 of 40 WSSV positive samples were successfully cloned and sequenced; among the ORF14/15 genes of 25 WSSV-positive strains, 22 was 619 bp in length and 3 was 620 bp. All the 25 Guangxi strains carried a 5949-bp deletion in the ORF14/15 region relative to TH-96-II, which has the longest nucleotide sequence in this region; the deletion of Guangxi strains occurred in the middle region of ORF14/15 gene, with only 190 bp and 429 bp/ 430 bp at 5' and 3' ends, respectively, which were coincident with WSSV-IN-05-I in deletion length and position. Sixteen of 25 Guangxi strains had completely identical nucleotide sequences in the variable re gion, and the homology between other strains also exceeded 97.9%. There were single nucleotide substi tution, deletion, and insertion in the ORF14/15 region of Guangxi strains compared with other strains in GenBank. In the phylogenetic tree based on WSSV variable region ORF14/15, the Guangxi strains were closely related and formed a separate branch with Indian strain IN-05-I, but far from other strains in GenBank. The ORF14/15 gene of WSSV isolates in cultured P. vannamei in Guangxi has a large deletion in the middle of the variable region, and the Guangxi WSSV strains show no significant spatio-temporal differences; the Guangxi strains are closer in genetics to Indian strain IN-05-I than other strains in GenBank. PMID:24772898

  8. The DNA Virus White Spot Syndrome Virus Uses an Internal Ribosome Entry Site for Translation of the Highly Expressed Nonstructural Protein ICP35

    Science.gov (United States)

    Kang, Shih-Ting; Wang, Han-Ching; Yang, Yi-Ting

    2013-01-01

    Although shrimp white spot syndrome virus (WSSV) is a large double-stranded DNA virus (?300 kbp), it expresses many polycistronic mRNAs that are likely to use internal ribosome entry site (IRES) elements for translation. A polycistronic mRNA encodes the gene of the highly expressed nonstructural protein ICP35, and here we use a dual-luciferase assay to demonstrate that this protein is translated cap independently by an IRES element located in the 5? untranslated region of icp35. A deletion analysis of this region showed that IRES activity was due to stem-loops VII and VIII. A promoterless assay, a reverse transcription-PCR together with quantitative real-time PCR analysis, and a stable stem-loop insertion upstream of the Renilla luciferase open reading frame were used, respectively, to rule out the possibility that cryptic promoter activity, abnormal splicing, or read-through was contributing to the IRES activity. In addition, a Northern blot analysis was used to confirm that only a single bicistronic mRNA was expressed. The importance of ICP35 to viral replication was demonstrated in a double-stranded RNA (dsRNA) interference knockdown experiment in which the mortality of the icp35 dsRNA group was significantly reduced. Tunicamycin was used to show that the ? subunit of eukaryotic initiation factor 2 is required for icp35 IRES activity. We also found that the intercalating drug quinacrine significantly inhibited icp35 IRES activity in vitro and reduced the mortality rate and viral copy number in WSSV-challenged shrimp. Lastly, in Sf9 insect cells, we found that knockdown of the gene for the Spodoptera frugiperda 40S ribosomal protein RPS10 decreased icp35 IRES-regulated firefly luciferase activity but had no effect on cap-dependent translation. PMID:24089551

  9. First report on White Spot Syndrome Virus (WSSV infection in white leg shrimp Litopenaeus vannamei (Crustacea, Penaeidae under semi intensive culture condition in India

    Directory of Open Access Journals (Sweden)

    Gunalan Balakrishnan

    2011-07-01

    Full Text Available Scientific shrimp culture began in India in the late eighties along the east coast particularly inAndrapradesh and Tamilnadu. Continuous success of shrimp culture was affected by mass mortalities ofcultured shrimp in 1994. Thereafter disease infection on survival and production of shrimps get itsimportance in culture. The present study is the first report on WSSV (white spot syndrome virusinfection in cultured Litopenaeus vannamei (Boone, 1931 in India. WSSV infection was observed on 70thdays of culture due to cross contamination of white spot infected shrimp from the neighboring farmbecause of birds. Due to this infection within two days the mortality ratio has gone up to 25% in pond 1and 12% in pond 2. So this present study strongly recommends to every shrimp farmers to go for birdfencing & crab fencing to avoid horizontal contamination, before stocking the good quality seed, thenthey will have the risk free WSSV culture.

  10. Influence of Agathi grandiflora active principles inhibit viral multiplication and stimulate immune system in Indian white shrimp Fenneropenaeus indicus against white spot syndrome virus infection.

    Science.gov (United States)

    Bindhu, Francis; Velmurugan, Subramanian; Donio, Mariathason Birdilla Selva; Michaelbabu, Mariavincent; Citarasu, Thavasimuthu

    2014-12-01

    Five herbs including Adathoda vasica, Agathi grandiflora, Leucas aspera, Psoralea corylifolia, and Quercus infectoria were selected to screen the antiviral and immunostimulant activity against white spot syndrome virus (WSSV) and Vibrio harveyi respectively using different organic polar and non-polar solvents. Based on the initial screening results, ethyl acetate and methanolic extracts of A. grandiflora had strong antiviral and immunostimulant activities. Those extracts incubated with WSSV injected Fenneropenaeus indicus got only 20% mortality and no PCR positive signals were seen in two step PCR amplification. The methanolic extracts of A. grandiflora were further purified through silica column chromatography and the fractions screened again for antiviral and immunostimulant activity. The secondary screening results revealed that, the fractions of F5 to F7 had effectively controlled the WSSV multiplication and V. harveyi growth. The pooled fractions (F5 to F7) was structurally characterized by gas chromatograph-mass spectrometry (GC-MS) analysis and few compounds were identified including 3,7.11,15-Tetramethyl-2-Hexane-1-ol, pytol and 1,2-Benzenedicarboxylic acid, diisooctyl ester. The pooled fractions were mixed with the basal feed ingredients at the concentration of 100 (D-1), 200 (D-2), 300 (D-3) and 400 (D-4) mg kg(-1) and the diets fed to the F. indicus (9.0 ± 0.5 g) for 30 days. After the completion of feeding trail, they were challenged with virulent WSSV and studied the cumulative mortality, molecular diagnosis by quantitative real time PCR (qRT-PCR), biochemical, haematological and immunological parameters. The control diet fed F. indicus succumbed to death 100% within 3 days whereas the D-3 and D-4 helped to reduced the cumulative mortality of 60-80% respectively. The qRT-PCR revealed that, the WSSV copy number was gradually decreased when increasing concentration of A. grandiflora extract active fraction in the diets. The diets D-3 and D-4 helped to reduce the protein and carbohydrate levels significantly (P WSSV challenge. PMID:25301717

  11. 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.

  12. 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.

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

    Scientific Electronic Library Online (English)

    Patricia, Braunig; Rafael Diego da, Rosa; Caroline Heidrich, Seibert; Mariana, Borsa; Patricia Hermes, Stoco; Edmundo Carlos, Grisard; Aguinaldo Roberto, Pinto.

    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.

  14. Protection of Penaeus monodon from Infection of White spot syndrome virus by DNA Construct Expressing Long Hairpin-RNA Against ICP11 Gene

    OpenAIRE

    Das, Rekha; Karthireddy, Syamala; Gireesh-Babu, P.; Reddy, A.K.; Krishna, Gopal; Chaudhari, Aparna

    2010-01-01

    A plasmid construct (pICP11-LH) was designed to constitutively express long-hairpin RNA (lhRNA) against icp11 gene, which is reportedly the most highly expressed gene of White spot syndrome virus (WSSV) and likely to have an important role in viral pathogenesis. The construct was used singly and in combination with other similar constructs designed against vp28 and vp19. A total of 6 treatments, T1 (pICP11-LH; 35 ?g), T2 (pVP28-LH; 35 ?g), T3 (pVP28-LH and pVP19-LH; 17.5 ?g each), T4 (pVP28-L...

  15. Identification of White Spot Syndrome Virus Latency-Related Genes in Specific-Pathogen-Free Shrimps by Use of a Microarray

    Science.gov (United States)

    Khadijah, Siti; Neo, Soek Ying; Hossain, M. S.; Miller, Lance D.; Mathavan, S.; Kwang, Jimmy

    2003-01-01

    To investigate whether specific-pathogen-free (SPF) shrimps are asymptomatic carriers of white spot syndrome virus (WSSV), we used a WSSV-specific DNA microarray to measure WSSV gene expression in SPF and WSSV-infected shrimps. Three WSSV genes were found to be relatively highly expressed in SPF shrimps. Reverse transcription-PCR using nested primers as well as real-time detection confirmed that these genes have no detectable counterparts in GenBank; structural analysis of the putative proteins revealed helix-loop-helix and leucine zipper motifs. Viral sequences could be PCR amplified from genomic DNA of SPF shrimp, further supporting the suggestion that these shrimps are asymptomatic carriers. PMID:12941929

  16. Validation of a Commercial Insulated Isothermal PCR-based POCKIT Test for Rapid and Easy Detection of White Spot Syndrome Virus Infection in Litopenaeus vannamei

    Science.gov (United States)

    Tsai, Yun-Long; Wang, Han-Ching; Lo, Chu-Fang; Tang-Nelson, Kathy; Lightner, Donald; Ou, Bor-Rung; Hour, Ai-Ling; Tsai, Chuan-Fu; Yen, Cheng-Chi; Chang, Hsiao-Fen Grace; Teng, Ping-Hua; Lee, Pei-Yu

    2014-01-01

    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 using700 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. PMID:24625894

  17. KLONING DAN SEQUENCING KERANGKA BACA TERBUKA VP26 DAN VP28 VIRUS WSSV SEBAGAI KANDIDAT VAKSIN UNTUK PENGENDALIAN PENYAKIT BINTIK PUTIH (WHITE SPOT DISEASE) PADA UDANG WINDU (PENAEUS MONODON)

    OpenAIRE

    Hilal Anshary; Triyanto; Gunarto Latama; Sriwulan

    2012-01-01

    White spot syndrome virus (WSSV) merupakan virus patogen yang paling mematikan bagi udang windu (Peneaus monodon) dan telah menimbulkan kerugian ekonomi yang sangat besar serta mengancam perkembangan industri budidaya udang di dunia termasuk di Indonesia. Genom WSSV telah ditentukan urutan nukleotidanya secara lengkap. Berbagai upaya telah digunakan untuk mengatasi masalah infeksi wssv, antara lain penerapan biosecurity, penerapan good aquaculture practices serta menebar benur udang yang berl...

  18. Susceptibility of juvenile European lobster Homarus gammarus to shrimp products infected with high and low doses of white spot syndrome virus.

    Science.gov (United States)

    Bateman, K S; Munro, J; Uglow, B; Small, H J; Stentiford, G D

    2012-08-27

    White spot syndrome virus (WSSV) is the most important pathogen known to affect the sustainability and growth of the global penaeid shrimp farming industry. Although most commonly associated with penaeid shrimp farmed in warm waters, WSSV is also able to infect, cause disease in and kill a wide range of other decapod crustaceans, including lobsters, from temperate regions. In 2005, the European Union imported US$500 million worth of raw frozen or cooked frozen commodity products, much of which originated in regions positive for white spot disease (WSD). The presence of WSSV within the UK food market was verified by means of nested PCR performed on samples collected from a small-scale survey of supermarket commodity shrimp. Passage trials using inoculum derived from commodity shrimp from supermarkets and delivered by injection to specific pathogen-free Pacific white shrimp Litopenaeus vannamei led to rapid mortality and pathognomonic signs of WSD in the shrimp, demonstrating that WSSV present within commodity shrimp was viable. We exposed a representative European decapod crustacean, the European lobster Homarus gammarus, to a single feeding of WSSV-positive, supermarket-derived commodity shrimp, and to positive control material (L. vannamei infected with a high dose of WSSV). These trials demonstrated that lobsters fed positive control (high dose) frozen raw products succumbed to WSD and displayed pathognomonic signs associated with the disease as determined by means of histology and transmission electron microscopy. Lobsters fed WSSV-positive, supermarket-derived commodity shrimp (low dose) did not succumb to WSD (no mortality or pathognomonic signs of WSD) but demonstrated a low level or latent infection via PCR. This study confirms susceptibility of H. gammarus to WSSV via single feedings of previously frozen raw shrimp products obtained directly from supermarkets. PMID:23186704

  19. White Spot Syndrome Virus vulnerability of Tiger shrimp (Penaeus monodon cultured in the coastal ponds of Cox’s Bazar region, Bangladesh

    Directory of Open Access Journals (Sweden)

    Mohammad M. Monwar

    2013-07-01

    Full Text Available Jumbo tiger prawn, Penaeus monodon is the world’s most popular cultivable species for itsfast growth, hardy nature, delicious taste and market demand. During the last three decades shrimpculture has been expanded rapidly. White spot Syndrome Virus (WSSV is a great treat to this cultureexpansion. In this research, WSSV free fries were stocked in four coastal ponds with different salinities.The stocking density was kept low and constant. Water parameters, fish growth, and WSSVcontamination were recorded fortnightly. The WSSV contamination was primarily detected by using EnbioShrimp Virus Detection Test Kit, ‘Shrimple’ and further confirmed by the PCR test. Among the ponds,Pond A, B, and C were in completely controlled environmental condition where as pond D was traditionalone that exposed to tidal variations. Physico-chemical parameter varied from as, temperature 29 to32ºC, salinity 0 to 31‰, water pH 7.1 to 8.3, dissolved oxygen 3.8 to 6.3 mL L-1, alkalinity 80-122 mg L-1,ammonia 0 to 1.5 mg L-1 and transparency 23 to 50 cm. The WSSV was found positive in the pond D justafter hundred days, when the average weight of the shrimps was 26.69 g. It is observed that rapidchange in the salinity and temperature, poor environmental conditions and uncontrolled exchange ofwater made the shrimp more vulnerable to the WSSV.

  20. White Spot Syndrome Virus IE1 and WSV056 Modulate the G1/S Transition by Binding to the Host Retinoblastoma Protein

    Science.gov (United States)

    Ran, Xiaozhuo; Bian, Xiaofang; Ji, Yongchang; Yan, Xiumin; Yang, Feng

    2013-01-01

    DNA viruses often target cellular proteins to modulate host cell cycles and facilitate viral genome replication. However, whether proliferation of white spot syndrome virus (WSSV) requires regulation of the host cell cycle remains unclear. In the present study, we show that two WSSV paralogs, IE1 and WSV056, can interact with Litopenaeus vannamei retinoblastoma (Rb)-like protein (lv-RBL) through the conserved LxCxE motif. Further investigation revealed that IE1 and WSV056 could also bind to Drosophila retinoblastoma family protein 1 (RBF1) in a manner similar to how they bind to lv-RBL. Using the Drosophila RBF-E2F pathway as a model system, we demonstrated that both IE1 and WSV056 could sequester RBF1 from Drosophila E2F transcription factor 1 (E2F1) and subsequently activate E2F1 to stimulate the G1/S transition. Our findings provide the first evidence that WSSV may regulate cell cycle progression by targeting the Rb-E2F pathway. PMID:24027329

  1. Transcriptional upregulation of fortilin in shrimp, Penaeus (Metapenaeus japonicus fed diets containing recombinant VP28, an antigenic protein of white spot syndrome virus (WSSV

    Directory of Open Access Journals (Sweden)

    Christopher Marlowe A. Caipang

    2013-07-01

    Full Text Available White spot syndrome virus (WSSV is considered as one of the serious viral pathogens ofshrimp. There are several preventive measures that have been developed to curb the devastating effectsof this virus in shrimp aquaculture. Juvenile shrimps, Penaeus (Metapenaeus japonicus were fed withcommercial feeds that were mixed with recombinant VP28, a structural protein antigen of WSSV for aperiod of 14 days. The immune response of the shrimp during oral administration of the medicated feedwas determined by expression analysis of fortilin, a gene that is involved in the antiviral response. Therewas a significant increase in the level of expression of fortilin both in the gut and the gills in the fedgroup during the duration of feeding. The level of expression gradually decreased in the fed group,whereby at the 7th and 14th day after the last day of feeding with medicated feeds, no significantdifferences were observed between the fed and control groups in the expression at the gills and gut,respectively.

  2. Expression profile of bio-defense genes in Penaeus monodon gills in response to formalin inactivated white spot syndrome virus vaccine.

    Science.gov (United States)

    Sudheer, N S; Poulose, Gigi; Thomas, Ancy; Viswanath, Kiron; Kulkarni, Amod; Narayanan, R B; Philip, Rosamma; Singh, I S Bright

    2015-05-01

    White spot syndrome virus (WSSV) is the most devastating pathogen of penaeid shrimp. While developing technology to vaccinate shrimp against WSSV, it is imperative to look into the immune response of the animal at molecular level. However, very little information has been generated in this direction. The present study is an attempt to understand the expression of bio-defense genes in gill tissues of Penaeus monodon in response to formalin inactivated WSSV. A WSSV vaccine with a viral titer of 1×10(9) DNA copies was prepared and orally administered to P. monodon at a rate of 1.75×10(6) DNA copies of inactivated virus preparation (IVP) day(-1) for 7days. The animals were challenged with WSSV on 1st and 5th day post vaccination, and temporal expression of bio-defense genes in gill tissues was studied. Survival of 100% and 50% were observed respectively on 1st and 5th day post vaccination challenge. The humoral immune genes prophenoloxidase (proPO), alpha 2-macroglobulin (?2M), crustin and PmRACK, and the cell mediated immune genes caspase and Rab7 were up regulated in gill tissue upon vaccination and challenge. The expression of humoral gene crustin and cellular gene Rab7 was related to survival in IVP administered shrimp. Results of the study suggest that these genes have roles in protecting shrimp from WSSV on vaccination. PMID:25735464

  3. Crystal Structures of Major Envelope Proteins VP26 and VP28 from White Spot Syndrome Virus Shed Light on Their Evolutionary Relationship?

    Science.gov (United States)

    Tang, Xuhua; Wu, Jinlu; Sivaraman, J.; Hew, Choy Leong

    2007-01-01

    White spot syndrome virus (WSSV) is a virulent pathogen known to infect various crustaceans. It has bacilliform morphology with a tail-like appendage at one end. The envelope consists of four major proteins. Envelope structural proteins play a crucial role in viral infection and are believed to be the first molecules to interact with the host. Here, we report the localization and crystal structure of major envelope proteins VP26 and VP28 from WSSV at resolutions of 2.2 and 2.0 Å, respectively. These two proteins alone account for approximately 60% of the envelope, and their structures represent the first two structural envelope proteins of WSSV. Structural comparisons among VP26, VP28, and other viral proteins reveal an evolutionary relationship between WSSV envelope proteins and structural proteins from other viruses. Both proteins adopt ?-barrel architecture with a protruding N-terminal region. We have investigated the localization of VP26 and VP28 using immunoelectron microscopy. This study suggests that VP26 and VP28 are located on the outer surface of the virus and are observed as a surface protrusion in the WSSV envelope, and this is the first convincing observation for VP26. Based on our studies combined with the literature, we speculate that the predicted N-terminal transmembrane region of VP26 and VP28 may anchor on the viral envelope membrane, making the core ?-barrel protrude outside the envelope, possibly to interact with the host receptor or to fuse with the host cell membrane for effective transfer of the viral infection. Furthermore, it is tempting to extend this host interaction mode to other structural viral proteins of similar structures. Our finding has the potential to extend further toward drug and vaccine development against WSSV. PMID:17409146

  4. 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.

  5. Protection of shrimp against white spot syndrome virus (WSSV) with ?-1,3-D-glucan-encapsulated vp28-siRNA particles.

    Science.gov (United States)

    Zhu, Fei; Zhang, Xiaobo

    2012-02-01

    White spot syndrome virus (WSSV) is a major shrimp viral pathogen responsible for large economic losses to shrimp aquaculture all over the world. The RNAi mediated by siRNA contributes a new strategy to control this viral disease. However, the efficient approach to deliver the siRNA into shrimp remains to be addressed. In this investigation, an antiviral vp28-siRNA was encapsulated in ?-1,3-D-glucan, and then the ?-1,3-D-glucan-encapsulated vp28-siRNA particles (GeRPs) were delivered into Marsupenaeus japonicus shrimp. The results showed that the vp28-siRNA in GeRPs could be released in hemocytes of shrimp. It was found that the GeRPs containing the vp28-siRNA inhibited the replication of WSSV in vivo, which presented a better antiviral activity than the non-encapsulated vp28-siRNA. Further evidence indicated that the mortality of WSSV-infected shrimp was significantly delayed by the GeRPs containing vp28-siRNA. Therefore, our study presented that the glucan-encapsulated siRNA might represent a novel potential therapeutic or preventive approach to control the shrimp disease. PMID:21590271

  6. PmVRP15, a Novel Viral Responsive Protein from the Black Tiger Shrimp, Penaeus monodon, Promoted White Spot Syndrome Virus Replication

    Science.gov (United States)

    Vatanavicharn, Tipachai; Prapavorarat, Adisak; Jaree, Phattarunda; Somboonwiwat, Kunlaya; Tassanakajon, Anchalee

    2014-01-01

    Suppression subtractive hybridization of Penaeus monodon hemocytes challenged with white spot syndrome virus (WSSV) has identified the viral responsive gene, PmVRP15, as the highest up-regulated gene ever reported in shrimps. Expression analysis by quantitative real time RT-PCR revealed 9410–fold up-regulated level at 48 h post WSSV injection. Tissue distribution analysis showed that PmVRP15 transcript was mainly expressed in the hemocytes of shrimp. The full-length cDNA of PmVRP15 transcript was obtained and showed no significant similarity to any known gene in the GenBank database. The predicted open reading frame of PmVRP15 encodes for a deduced 137 amino acid protein containing a putative transmembrane helix. Immunofluorescent localization of the PmVRP15 protein revealed it accumulated around the nuclear membrane in all three types of shrimp hemocytes and that the protein was highly up-regulated in WSSV-infected shrimps. Double-stranded RNA interference-mediated gene silencing of PmVRP15 in P. monodon significantly decreased WSSV propagation compared to the control shrimps (injected with GFP dsRNA). The significant decrease in cumulative mortality rate of WSSV-infected shrimp following PmVRP15 knockdown was observed. These results suggest that PmVRP15 is likely to be a nuclear membrane protein and that it acts as a part of WSSV propagation pathway. PMID:24637711

  7. Immunogenicity and protective efficacy of a major White Spot Syndrome Virus (WSSV) envelope protein VP24 expressed in Escherichia coli against WSSV.

    Science.gov (United States)

    Thomas, Ancy; Sudheer, Naduvilamuriparampu Saidumuhammed; Viswanathan, Karthik; Kiron, Viswanath; Bright Singh, Issac S; Narayanan, Rangarajan Badri

    2014-11-01

    The study reports cloning, expression and characterization of immunogenic activity of VP24, a major envelope protein of White Spot Syndrome Virus (WSSV). His-tagged VP24 was expressed as truncated protein and purified from inclusion bodies by metal affinity chromatography under denaturing conditions. The ability to confer protection from WSSV by oral administration of recombinant viral protein (rVP24) was examined in black tiger shrimp Penaeus monodon (P. monodon) juveniles (advanced post larvae). Animals were fed with rVP24 for 10 days, orally challenged with WSSV and assayed for expression of viral genes and shrimp immune genes on the 2nd, 5th and 8th days of challenge. The survival of juvenile shrimps in the vaccinated and challenged group was significantly higher compared to the unvaccinated and challenged group with lesser viral gene expression (DNA polymerase, latency 1 and vp28). Analysis of immune gene expression showed upregulation of syntenin and down regulation of STAT, Rab 7 and caspase during the experimental period. This study points to the feasibility of using rVP24 as candidate vaccine in P. monodon against WSSV. PMID:25218401

  8. The promoter of the white spot syndrome virus immediate-early gene WSSV108 is activated by the cellular KLF transcription factor.

    Science.gov (United States)

    Liu, Wang-Jing; Lo, Chu-Fang; Kou, Guang-Hsiung; Leu, Jiann-Horng; Lai, Ying-Jang; Chang, Li-Kwan; Chang, Yun-Shiang

    2015-03-01

    A series of deletion and mutation assays of the white spot syndrome virus (WSSV) immediate-early gene WSSV108 promoter showed that a Krüppel-like factor (KLF) binding site located from -504 to -495 (relative to the transcription start site) is important for the overall level of WSSV108 promoter activity. Electrophoretic mobility shift assays further showed that overexpressed recombinant Penaeus monodon KLF (rPmKLF) formed a specific protein-DNA complex with the (32)P-labeled KLF binding site of the WSSV108 promoter, and that higher levels of Litopenaeus vannamei KLF (LvKLF) were expressed in WSSV-infected shrimp. A transactivation assay indicated that the WSSV108 promoter was strongly activated by rPmKLF in a dose-dependent manner. Lastly, we found that specific silencing of LvKLF expression in vivo by dsRNA injection dramatically reduced both WSSV108 expression and WSSV replication. We conclude that shrimp KLF is important for WSSV genome replication and gene expression, and that it binds to the WSSV108 promoter to enhance the expression of this immediate-early gene. PMID:25445906

  9. Regulation of the immediate-early genes of white spot syndrome virus by Litopenaeus vannamei kruppel-like factor (LvKLF).

    Science.gov (United States)

    Huang, Ping-Han; Lu, Shao-Chia; Yang, Shu-Han; Cai, Pei-Si; Lo, Chu-Fang; Chang, Li-Kwan

    2014-10-01

    Kruppel-like factors (KLFs) belong to a subclass of Cys2/His2 zinc-finger DNA-binding proteins, and act as important regulators with diverse roles in cell growth, proliferation, differentiation, apoptosis and tumorigenesis. Our previous research showed that PmKLF from Penaeus monodon is crucial for white spot syndrome virus (WSSV) infection, yet the mechanisms by which PmKLF influences WSSV infection remain unclear. This study cloned KLF from Litopenaeus vannamei (LvKLF), which had 93% similarity with PmKLF. LvKLF formed a dimer via the C-terminal zinc-finger motif. Knockdown of LvKLF expression by dsRNA injection in WSSV-challenged shrimps was found to significantly inhibit the transcription of two important immediate-early (IE) genes, IE1 and WSSV304, and also reduced WSSV copy numbers. Moreover, reporter assays revealed that the promoter activities of these two WSSV IE genes were substantially enhanced by LvKLF. Mutations introduced in the promoter sequences of IE1 and WSSV304 were shown to abolish LvKLF activation of promoter activities; and an electrophoretic mobility shift assay demonstrated that LvKLF binds to putative KLF-response elements (KRE) in the promoters. Taken together, these results indicate that LvKLF transcriptional regulation of key IE genes is critical to WSSV replication. PMID:24881625

  10. Generation of recombinant monoclonal antibodies to study structure-function of envelope protein VP28 of white spot syndrome virus from shrimp

    International Nuclear Information System (INIS)

    White spot syndrome virus (WSSV) is a major pathogen in shrimp aquaculture. VP28 is one of the most important envelope proteins of WSSV. In this study, a recombinant antibody library, as single-chain fragment variable (scFv) format, displayed on phage was constructed using mRNA from spleen cells of mice immunized with full-length VP28 expressed in Escherichia coli. After several rounds of panning, six scFv antibodies specifically binding to the epitopes in the N-terminal, middle, and C-terminal regions of VP28, respectively, were isolated from the library. Using these scFv antibodies as tools, the epitopes in VP28 were located on the envelope of the virion by immuno-electron microscopy. Neutralization assay with these antibodies in vitro suggested that these epitopes may not be the attachment site of WSSV to host cell receptor. This study provides a new way to investigate the structure and function of the envelope proteins of WSSV

  11. Identification of a Novel Nonstructural Protein, VP9, from White Spot Syndrome Virus: Its Structure Reveals a Ferredoxin Fold with Specific Metal Binding Sites?

    Science.gov (United States)

    Liu, Yang; Wu, Jinlu; Song, Jianxing; Sivaraman, J.; Hew, Choy L.

    2006-01-01

    White spot syndrome virus (WSSV) is a major pathogen in shrimp aquaculture. VP9, a full-length protein of WSSV, encoded by open reading frame wsv230, was identified for the first time in the infected Penaeus monodon shrimp tissues, gill, and stomach as a novel, nonstructural protein by Western blotting, mass spectrometry, and immunoelectron microscopy. Real-time reverse transcription-PCR demonstrated that the transcription of VP9 started from the early to the late stage of WSSV infection as a major mRNA species. The structure of full-length VP9 was determined by both X-ray and nuclear magnetic resonance (NMR) techniques. It is the first structure to be reported for WSSV proteins. The crystal structure of VP9 revealed a ferredoxin fold with divalent metal ion binding sites. Cadmium sulfate was found to be essential for crystallization. The Cd2+ ions were bound between the monomer interfaces of the homodimer. Various divalent metal ions have been titrated against VP9, and their interactions were analyzed using NMR spectroscopy. The titration data indicated that VP9 binds with both Zn2+ and Cd2+. VP9 adopts a similar fold as the DNA binding domain of the papillomavirus E2 protein. Based on our present investigations, we hypothesize that VP9 might be involved in the transcriptional regulation of WSSV, a function similar to that of the E2 protein during papillomavirus infection of the host cells. PMID:16956937

  12. The shrimp IKK-NF-?B signaling pathway regulates antimicrobial peptide expression and may be subverted by white spot syndrome virus to facilitate viral gene expression.

    Science.gov (United States)

    Wang, Pei-Hui; Gu, Zhi-Hua; Wan, Ding-Hui; Liu, Bo-Du; Huang, Xian-De; Weng, Shao-Ping; Yu, Xiao-Qiang; He, Jian-Guo

    2013-09-01

    The I?B kinases IKK? and IKK? and the IKK-related kinases TANK-binding kinase 1 (TBK1) and IKK? are the master regulators of the NF-?B signaling pathway. Although this pathway has been extensively studied in mammals, less attention has been paid in crustaceans, which have significant economic value. Here, we report the cloning and functional studies of two IKK homologs, LvIKK? and LvIKK?, from Pacific white shrimp, Litopenaeus vannamei. LvIKK? and LvIKK? mRNAs are widely expressed in different tissues and are responsive to white spot syndrome virus (WSSV) infection. When overexpressed in Drosophila S2 cells, LvIKK? but not LvIKK? activates the promoters of NF-?B pathway-controlled antimicrobial peptide genes (AMPs), such as the Penaeidins (PENs). In HEK 293T cells, both LvIKK? and LvIKK? activate an NF-?B reporter. The silencing of LvIKK? or LvIKK? using double-stranded RNA (dsRNA)-mediated RNA interference (RNAi) decreases the expression of L. vannamei AMPs, including PENs, lysozyme and crustins. Intriguingly, LvIKK?- or LvIKK?-silenced L. vannamei are resistant to WSSV infection. We hypothesized that successful infection with WSSV requires the activation of the IKK-NF-?B signaling pathway to modulate viral gene expression. We constructed luciferase reporters for 147 WSSV genes. By screening, we found that the WSV051, WSV059, WSV069, WSV083, WSV090, WSV107, WSV244, WSV303, WSV371 and WSV445 promoters can be activated by LvIKK? or LvIKK? in Drosophila S2 cells. Taken together, our results reveal that LvIKK? and LvIKK? may participate in the regulation of shrimp AMPs and that WSSV may subvert the L. vannamei IKK-NF-?B signaling pathway to facilitate viral gene expression. PMID:23954949

  13. 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. PMID:25912357

  14. Astakine LvAST binds to the ? subunit of F1-ATP synthase and likely plays a role in white shrimp Litopeneaus vannamei defense against white spot syndrome virus.

    Science.gov (United States)

    Liang, Gao-Feng; Liang, Yan; Xue, Qinggang; Lu, Jin-Feng; Cheng, Jun-Jun; Huang, Jie

    2015-03-01

    Cytokines play a critical role in innate and adaptive immunity. Astakines represent a group of invertebrate cytokines that are related to vertebrate prokineticin and function in promoting hematopoiesis in crustaceans. We have identified an astakine from the white shrimp Litopeneaus vannamei and named it LvAST in a previous research. In the present research, we investigated the interactions among LvAST, the envelope protein VP37 of white spot syndrome virus (i.e., WSSV), and the ? subunit of F1-ATP synthase (ATPsyn-?) of the white shrimp (i.e., BP53) using binding assays and co-precipitations. We also examined the effects of LvAST on shrimp susceptibility to WSSV. We found that LvAST and VP37 competitively bound to BP53, but did not bind to each other. Shrimps that had been injected with recombinant LvAST exhibited significantly lower mortality and longer survival time in experimental infections by WSSV. In contrast, shrimps whose LvAST gene expression had been inhibited by RNA interference showed significantly higher WSSV infection intensity and shorter survival time following viral challenges. These results suggested that LvAST and WSSV both likely use ATPsyn-? as a receptor and LvAST plays a role in shrimp defense against WSSV infection. This represented the first research showing the involvement of astakines in host antiviral immunity. PMID:25536411

  15. Detection of White Spot Syndrome Virus in Brazil using Negative Staining, Immunoelectron Microscopy and Immunocytochemistry Techniques / Detección del Virus del Síndrome de Mancha Blanca en el Brasil Utilizando Inmunomicroscopía e Inmunomarcación con Partículas de Oro Coloidal

    Scientific Electronic Library Online (English)

    M, Hipolito; M. H. B, Catroxo; A. M. C. R. P. F, Martins; N.A, Melo; E. M, Pituco; N.T.C, Galleti; M. J. T, Ranzani-Paiva; J.L.P, Mouriño; C.M, Ferreira.

    2012-06-01

    Full Text Available Se obtuvieron para el estudio 30 muestras de camarones marinos comerciales (L. vannamei) de las granjas de la región sur de Brasil. Fueron procesados fragmentos de hepatopáncreas y raspados internos del cefalotórax recogidos en estos animales por microscopía electrónica de transmisión con tinción ne [...] gativa (preparación rápida), inmunomicroscopía y técnicas de inmunocitoquímica (inmunomarcación con partículas de oro coloidal). En la microscopía electrónica de transmisión de un gran número de partículas de virus de la mancha blanca, ovoide o elipsoidal a baciliformes, medían 230-290 nm de longitud y 80-160 nm de diámetro. En 27 (90%) de las 30 muestras examinadas intra-nuclear proyecciones se visualizaron mediante la técnica de tinción negativa. Utilizando una técnica de inmunomicroscopía electrónica, el anti-suero VP 664 reunió a un gran número de partículas formadas por la interacción antígeno-anticuerpo. En la técnica de inmunocitoquímica, la reacción antígeno-anticuerpo fue fuertemente reforzada por las partículas de oro coloidal en los virus. En particular, en Brasil este es el primer informe, a nuestro entender, que describe el uso de estas técnicas de microscopía en muestras de camarón marino L. vanamei. Además, esta metodología también parece ser una herramienta complementaria viable para diagnosticar la presencia del virus de la mancha blanca en tejidos de camarón. Es importante destacar que estas son las primeras fotos en microscopia electrónica del WSSV obtenidas en Brasil. Abstract in english In this study thirty shrimp samples from commercial marine shrimp (L. vannamei) farms of southern region of Brazil were obtained. Hepatopancreas and shell scrapings fragments collected in these animals were processed by transmission electron microscopy using negative staining (rapid preparation), im [...] munoelectron microscopy and immunocytochemistry (immunolabelling with colloidal gold particles) techniques. On the transmission electron microscopy a great number of white spot virus particles, ovoid or bacilliform-to-ellipsoid, measured 230-290 nm in length and 80-160 nm in diameter with intra-nuclear projections were visualized by the negative staining technique in 27 (90%) out of 30 samples examined. Using immunoelectron microscopy technique, the anti-VP 664 serum agllutinated a large number of particles formed by antigen-antibody interaction. In the immunocytochemistry technique, the antigen-antibody reaction was styrongly marked by the particles of colloidal gold over the virus. Notably, this is the first report, to our knowledge, describing use of these microscopy techniques to study Brazilian L. vannamei marine shrimp samples; moreover, this methodology also appears to be a viable complementary tool for diagnosing the presence of the white spot virus within shrimp tissues. Importantly, these are the first photoelectron micrographs of the WSSV in Brazil.

  16. Detection of White Spot Syndrome Virus in Brazil using Negative Staining, Immunoelectron Microscopy and Immunocytochemistry Techniques Detección del Virus del Síndrome de Mancha Blanca en el Brasil Utilizando Inmunomicroscopía e Inmunomarcación con Partículas de Oro Coloidal

    Directory of Open Access Journals (Sweden)

    M Hipolito

    2012-06-01

    Full Text Available In this study thirty shrimp samples from commercial marine shrimp (L. vannamei farms of southern region of Brazil were obtained. Hepatopancreas and shell scrapings fragments collected in these animals were processed by transmission electron microscopy using negative staining (rapid preparation, immunoelectron microscopy and immunocytochemistry (immunolabelling with colloidal gold particles techniques. On the transmission electron microscopy a great number of white spot virus particles, ovoid or bacilliform-to-ellipsoid, measured 230-290 nm in length and 80-160 nm in diameter with intra-nuclear projections were visualized by the negative staining technique in 27 (90% out of 30 samples examined. Using immunoelectron microscopy technique, the anti-VP 664 serum agllutinated a large number of particles formed by antigen-antibody interaction. In the immunocytochemistry technique, the antigen-antibody reaction was styrongly marked by the particles of colloidal gold over the virus. Notably, this is the first report, to our knowledge, describing use of these microscopy techniques to study Brazilian L. vannamei marine shrimp samples; moreover, this methodology also appears to be a viable complementary tool for diagnosing the presence of the white spot virus within shrimp tissues. Importantly, these are the first photoelectron micrographs of the WSSV in Brazil.Se obtuvieron para el estudio 30 muestras de camarones marinos comerciales (L. vannamei de las granjas de la región sur de Brasil. Fueron procesados fragmentos de hepatopáncreas y raspados internos del cefalotórax recogidos en estos animales por microscopía electrónica de transmisión con tinción negativa (preparación rápida, inmunomicroscopía y técnicas de inmunocitoquímica (inmunomarcación con partículas de oro coloidal. En la microscopía electrónica de transmisión de un gran número de partículas de virus de la mancha blanca, ovoide o elipsoidal a baciliformes, medían 230-290 nm de longitud y 80-160 nm de diámetro. En 27 (90% de las 30 muestras examinadas intra-nuclear proyecciones se visualizaron mediante la técnica de tinción negativa. Utilizando una técnica de inmunomicroscopía electrónica, el anti-suero VP 664 reunió a un gran número de partículas formadas por la interacción antígeno-anticuerpo. En la técnica de inmunocitoquímica, la reacción antígeno-anticuerpo fue fuertemente reforzada por las partículas de oro coloidal en los virus. En particular, en Brasil este es el primer informe, a nuestro entender, que describe el uso de estas técnicas de microscopía en muestras de camarón marino L. vanamei. Además, esta metodología también parece ser una herramienta complementaria viable para diagnosticar la presencia del virus de la mancha blanca en tejidos de camarón. Es importante destacar que estas son las primeras fotos en microscopia electrónica del WSSV obtenidas en Brasil.

  17. The novel white spot syndrome virus-induced gene, PmERP15, encodes an ER stress-responsive protein in black tiger shrimp, Penaeus monodon.

    Science.gov (United States)

    Leu, Jiann-Horng; Liu, Kuan-Fu; Chen, Kuan-Yu; Chen, Shu-Hwa; Wang, Yu-Bin; Lin, Chung-Yen; Lo, Chu-Fang

    2015-04-01

    By microarray screening, we identified a white spot syndrome virus (WSSV)-strongly induced novel gene in gills of Penaeus monodon. The gene, PmERP15, encodes a putative transmembrane protein of 15?kDa, which only showed some degree of similarity (54-59%) to several unknown insect proteins, but had no hits to shrimp proteins. RT-PCR showed that PmERP15 was highly expressed in the hemocytes, heart and lymphoid organs, and that WSSV-induced strong expression of PmERP15 was evident in all tissues examined. Western blot analysis likewise showed that WSSV strongly up-regulated PmERP15 protein levels. In WSSV-infected hemocytes, immunofluorescence staining showed that PmERP15 protein was colocalized with an ER enzyme, protein disulfide isomerase, and in Sf9 insect cells, PmERP15-EGFP fusion protein colocalized with ER -Tracker™ Red dye as well. GRP78, an ER stress marker, was found to be up-regulated in WSSV-infected P.?monodon, and both PmERP15 and GRP78 were up-regulated in shrimp injected with ER stress inducers tunicamycin and dithiothreitol. Silencing experiments showed that although PmERP15 dsRNA-injected shrimp succumbed to WSSV infection more rapidly, the WSSV copy number had no significant changes. These results suggest that PmERP15 is an ER stress-induced, ER resident protein, and its induction in WSSV-infected shrimp is caused by the ER stress triggered by WSSV infection. Furthermore, although PmERP15 has no role in WSSV multiplication, its presence is essential for the survival of WSSV-infected shrimp. PMID:25499032

  18. Immune responses of whiteleg shrimp, Litopenaeus vannamei (Boone, 1931), to bacterially expressed dsRNA specific to VP28 gene of white spot syndrome virus.

    Science.gov (United States)

    Taju, G; Madan, N; Abdul Majeed, S; Kumar, T Raj; Thamizhvanan, S; Otta, S K; Sahul Hameed, A S

    2015-05-01

    In this study, dsRNA specific to VP28 gene of white spot syndrome virus (WSSV) of shrimp was synthesized in Escherichia coli in large scale and studied the immune response of shrimp to dsRNA-VP28. The haematological parameters such as clotting time and total haemocytes counts, and immunological parameters such as prophenoloxidase (proPO), superoxide dismutase (SOD), superoxide anion (SOA) and malondialdehyde content, as well as the mRNA expression of ten immune-related genes were examined to estimate the effect of dsRNA-VP28 on the innate immunity of Litopenaeus vannamei. The activities of proPO, SOA and SOD significantly increased in haemocyte after dsRNA-VP28 treatment, whereas MDA content did not change significantly. Among the ten immune-related genes examined, only the mRNA expression of proPO, cMnSOD, haemocyanin, crustin, BGBP, lipopolysaccharides (LPs), lectin and lysozyme in haemocytes, gill and hepatopancreas of L. vannamei, was significantly upregulated at 12 h after dsRNA-VP28 treatment, while no significant expression changes were observed in Toll receptor and tumour receptor genes. The increase of proPO and SOD activities, and SOA level and mRNA expression level of proPO, cMnSOD, haemocyanin, crustin, BGBP, LPs, lectin and lysozyme after dsRNA-VP28 stimulation indicate that these immune-related genes were involved in dsRNA-VP28-induced innate immunity in shrimp. PMID:24917208

  19. The protection of CpG ODNs and Yarrowia lipolytica harboring VP28 for shrimp Litopenaeus vannamei against White spot syndrome virus infection

    OpenAIRE

    Yi, Q.; Liu, R; Sun, R.; Wang, L (Lili); Z. Zhou; Wang, M.; Liu, Y.(Department of Modern Physics, University of Science and Technology of China, Hefei, Anhui, China); J Sun; C Madzak; Song, L

    2014-01-01

    The white spot syndrome is one of the most serious disease which has caused high mortalities and huge economic losses to shrimp culture. In the present study, the oral administrations with CpG ODNs and Yarrowia lipolytica harboring VP28 (rVP28-yl) as dietary supplement for shrimp Litopenaeus vannamei were conducted to evaluate their protective effects against WSSV. After feeding for 15 days, the cumulative mortality and the copy number of WSSV in CpG and rVP28-yl feeding shrimps were signific...

  20. Dietary administration of Gynura bicolor (Roxb. Willd.) DC water extract enhances immune response and survival rate against Vibrio alginolyticus and white spot syndrome virus in white shrimp Litopeneaus vannamei.

    Science.gov (United States)

    Wu, Chih-Chung; Chang, Yueh-Ping; Wang, Jyh-Jye; Liu, Chun-Hung; Wong, Saou-Lien; Jiang, Chii-Ming; Hsieh, Shu-Ling

    2015-01-01

    Gynura bicolor (Roxb. & Willd.) DC., a perennial plant belonging to the Asteraceae family, is originated from the tropical area of Asia. The total hemocyte count (THC), phenoloxidase (PO) activity, respiratory bursts (RBs), superoxide dismutase (SOD) activity, and lysozyme activity were examined after white shrimp Litopenaeus vannamei had been fed diets containing the water extract of G. bicolor at 0 (control), 0.5, 1.0, and 2.0 g (kg diet)(-1) for 7-28 days. The results indicated that these parameters increased accordingly with the amount of extract and time. THCs of the shrimp fed the G. bicolor diets at 1.0 and 2.0 g (kg diet)(-1) were significantly higher than that fed the control diet for 14-28 days. For the shrimp fed the G. bicolor diets at 0.5, 1.0, and 2.0 g (kg diet)(-1), the PO, RBs, and lysozyme activities reached the highest levels after 7 days, whereas SOD activity reached the highest levels after 14 days. In a separate experiment, white shrimp L. vannamei fed the diets containing the G. bicolor extract for 28 days were challenged with Vibrio alginolyticus at 3 × 10(6) cfu shrimp(-1) and white spot syndrome virus (WSSV) at 1 × 10(3) copies shrimp(-1). The survival rate of the shrimp fed the G. bicolor diets was significantly higher than that of the shrimp fed the control diet at 48-144 h post challenge V. alginolyticus and WSSV. For the shrimp fed the G. bicolor diets at 0.5, 1 and 2 g (kg diet)(-1) under challenges of V. alginolyticus and WSSV, their LPS- and ?-1,3-glucan-binding protein (LGBP) and peroxinectin (PE) mRNA expressions were significantly higher than those of the challenged control shrimp at 12-96 and 24-144 h post-challenge, respectively. We concluded that dietary administration of a G. bicolor extract could enhance the innate immunity within 28 days as evidenced by the increases in immune parameters (PO, RBs, and lysozyme) and antioxidant enzyme (SOD) activities of shrimp to against V. alginolyticus and WSSV infections. PMID:25462462

  1. Revisión de patogénesis y estrategias moleculares contra el virus del síndrome de la mancha blanca en camarones peneidos / A review of pathogenesis and molecular strategies against white spot syndrome virus of penaeid shrimp

    Scientific Electronic Library Online (English)

    Martin I, Bustillo-Ruiz; César M, Escobedo-Bonilla; Rogerio R, Sotelo-Mundo.

    2009-04-01

    Full Text Available RESUMEN El virus del síndrome de mancha blanca (WSSV) provoca graves mortandades en granjas de cultivo de camarones peneidos y serias pérdidas económicas. La secuencia y estructura genética excepcionales de WSSV lo colocan en su propia nueva familia, Nimaviridae. Recientemente, novedosas técnicas mo [...] leculares de alto rendimiento han permitido identificar y caracterizar varias proteínas estructurales de WSSV. Estas incluyen la secuenciación por `shotgun' y marcadores isobáricos para cuantificación absoluta y relativa (iTRAQ). Dichas técnicas han permitido caracterizar 14 nuevas proteínas de WSSV. La caracterización y localización de proteínas estructurales pueden ayudar a conocer la morfogénesis y patogénesis de WSSV. Ambos procesos son esenciales para entender el mecanismo de infección y para desarrollar nuevos métodos de control. Hasta ahora no existen tratamientos efectivos para combatir este virus en campo. Proteínas estructurales de WSSV como VP28 y VP19 han sido evaluadas para reducir el impacto de WSSV. Estas moléculas son esenciales en las etapas tempranas de infección. Bioensayos de neutralización usando anticuerpos específicos contra proteínas estructurales de WSSV han aumentado la supervivencia de camarones tratados. Recientemente, construcciones de RNA de interferencia (RNAi) dirigidos contra proteínas estructurales han sido usadas como una nueva herramienta para reducir/inhibir la replicación de WSSV. Una mejor comprensión de las interacciones hospedero-patógeno permitirá desarrollar nuevos métodos para controlar este virus. La localización y función de proteínas estructurales usando métodos de alto rendimiento contribuirá a implementar nuevas estrategias contra la infección. Métodos de intervención para bloquear la entrada del virus a la célula podrían ser valiosos productos de este tipo de investigaciones. Abstract in english ABSTRACT White spot syndrome virus (WSSV) causes high mortality to farmed shrimp and serious economic losses. Its unique sequence and genome structure has placed WSSV in its own new family Nimaviridae. Recently, high performance molecular techniques have made it possible to identify and characterize [...] several WSSV structural proteins. These include `shotgun' sequencing and isobaric tags for relative and absolute quantification (iTRAQ). Such techniques have made it possible to characterize 14 new WSSV proteins. Location and characterization of structural proteins can help to understand WSSV morphogenesis and pathogenesis. Both processes are essential to understand the mechanism of infection and to develop novel control methods. At present no effective treatments exist to fight WSSV in the field. WSSV structural proteins such as VP28 and VP19 have been evaluated to reduce the impact of WSSV. These molecules are essential early in the infection. Neutralization assays using specific antibodies against WSSV structural proteins have shown an increased survival of treated shrimp. Recently, RNA interference (RNAi) constructs directed against structural proteins have been used as a new tool to reduce/inhibit WSSV replication. A better comprehension of the host-pathogen interaction would allow the development of new methods to control WSSV. The use of high throughput techniques to determine the location and function of structural proteins will contribute to develop new strategies against infection. Intervention strategies aimed to block virus entry into the host cells may be a valuable output from these studies.

  2. The protection of CpG ODNs and Yarrowia lipolytica harboring VP28 for shrimp Litopenaeus vannamei against White spot syndrome virus infection

    Directory of Open Access Journals (Sweden)

    Q Yi

    2014-04-01

    Full Text Available The white spot syndrome is one of the most serious disease which has caused high mortalities and huge economic losses to shrimp culture. In the present study, the oral administrations with CpG ODNs and Yarrowia lipolytica harboring VP28 (rVP28-yl as dietary supplement for shrimp Litopenaeus vannamei were conducted to evaluate their protective effects against WSSV. After feeding for 15 days, the cumulative mortality and the copy number of WSSV in CpG and rVP28-yl feeding shrimps were significantly lower when they were challenged by WSSV, compared with those in control shrimps (p < 0.05. The caspase-3 activity was suppressed in rVP28-yl feeding shrimps but ascended in CpG feeding shrimps after WSSV challenge. Besides, the PO activity in CpG feeding shrimps was significantly increased after feeding trial, and kept increasing post WSSV challenge (p < 0.05. While the increased NO production was observed both in CpG and rVP28-yl feeding shrimps after feeding trial and WSSV challenge. In addition, increased mRNA expression levels of STAT and Dicer were observed in CpG group post WSSV challenge. These results together indicated that oral feeding of CpG ODNs and rVP28-yl could enhance the innate non-specific immune responses especially antiviral immunity of shrimps in varying degrees, and increase their resistance against WSSV infection

  3. Sustainable control of white spot disease

    DEFF Research Database (Denmark)

    Heinecke, Rasmus Demuth; Buchmann, Kurt

    White spot disease caused by the ciliate Ichthyophthirius multifiliis Fouquet, 1876 is a serious problem in freshwater aquaculture worldwide. This parasitosis is of frequent occurrence in both conventional earth pond fish farms and in fish farms using new high technology re-circulation systems. Chemicals such as formaldehyde, sodium percarbonate and sodium chloride have been applied to control infections with this parasite. However, information on the influence of concentration, length of treatment period and temperature, on the survival of theronts and tomonts, has been lacking. The tolerance of the free-living theront stage towards formaldehyde and sodium percarbonate in solution was investigated at two temperature levels (11-12 and 21-22 °C). The theronts were exposed to the two substances in concentrations of 8, 16, 32 and 64 mg/l. The survival of theronts was examined in a four-well setup. Each well contained 5-12 theronts in 150 µl aqueous solution. Tolerance towards formaldehyde and sodium percarbonate was tested simultaneously, together with two pure water negative controls. At each concentration and temperature (11-12 and 21-22 °C) the experiment was repeated 4-9 times. The numbers of live theronts were counted every 15 minutes using a stereo-microscope (8-100× magnification). Lysed and/or immobilised theronts with no movement of cilia were considered dead. For both substances a negative relationship was seen between the survival of theronts and length of treatment, temperature and concentration of chemical. Using formaldehyde or sodium percarbonate, all theronts were dead within 15 minutes at a concentration of 64 mg/l at 21-22 °C. At the same temperature, using 8 mg/l, this was achieved within 150 minutes using formaldehyde and 135 minutes using sodium percarbonate. At the low temperature (11-12 °C) using 64 mg/l, all theronts were dead within 45 minutes using formaldehyde and 30 minutes using sodium percarbonate. At 8 mg/l all theronts died within 300 minutes when treating with formaldehyde or sodium percarbonate. Dose-response experiments on tomonts showed that this parasite stage was much more tolerant towards both chemicals. Filtration experiments on tomont containing water were conducted. Mesh sizes of 500, 300, 160 and 80 µm were used. A mesh size of 80 µm was demonstrated to filter out 100 % of the tomonts. The abiotic factors temperature, chemical and concentration all had significant influences on parasite survival. Strategic treatment using the environmentally friendly chemical sodium percarbonate in combination with continuous micro-filtering of the pond water is suggested for control and management of this parasitosis.

  4. 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. PMID:25320443

  5. Presença do vírus da síndrome da mancha branca em crustáceos decápodes silvestres em lagoas costeiras no Sul do Brasil / Presence of the white spot syndrome virus (WSSV) in wild decapods crustaceans in coastal lagoons in southern Brazil

    Scientific Electronic Library Online (English)

    S.W., Costa; A.P.M., Fraga; A.S., Zamparetti; M.R.F., Marques; E.R., Andreatta.

    2012-02-01

    Full Text Available A presença do vírus da síndrome da mancha branca (em inglês WSSV) nas principais espécies de camarões, siris e caranguejos de cinco lagoas que recebem o efluente de fazendas afetadas pela enfermidade foi detectada por nested PCR, e inclusões virais nos camarões por histologia. Pela nested PCR encont [...] rou-se a presença de WSSV em 13 de 16 (81,2%) amostras de camarões da espécie Farfantepenaeus paulensis, em 13 de 14 (92,8%) de Litopenaeus schmitti, em uma de duas de Farfantepenaeus brasiliensis (50%), em 13 de 15 (86,6%) de siri da espécie Callinectes danae e em 11 de 12 (91,6%) de Callinectes sapidus, e não foi detectada no caranguejo Chasmagnathus granulata em 10 amostras. Inclusões características de WSSV foram observadas em três amostras histológicas de 50 (6,0%) no epitélio gástrico e cuticular e nas brânquias de dois exemplares de F. paulensis e um de L. schmitti. É o primeiro relato da presença de WSSV em camarões L. schmitti e no siri C. danae silvestres. As principais espécies de camarões e siris dos ambientes de entorno das fazendas foram contaminadas pelo WSSV, constituindo-se em vetores potenciais do vírus. Abstract in english The presence of white spot syndrome virus (WSSV) in the main species of shrimps, blue crabs, and burrowing crabs of five lagoons where shrimp farm effluents are discharged, was analyzed by nested PCR and the presence of virus inclusions in the shrimps was analyzed through histopathology. The nested [...] PCR analysis indicated the presence of WSSV in 13 of 16 (81.2%) samples of the shrimp species of Farfantepenaeus paulensis, in 13 of 14 (92.8%) of Litopenaeus schmitti, in one of two of Farfantepenaeus brasiliensis (50%), in 13 of 15 (86.6%) of blue crab species of Callinectes danae and in 11 of 12 (91.6%) of Callinectes sapidus and none was detected in the 10 samples of the burrowing crab Chasmagnathus granulata. The inclusion characteristics of WSSV were observed in three samples of 50 (6.0%) in the gastric and cuticular epithelium and in the gills of two specimens of F. paulensis and one of L. schmitti. The presence of WSSV in L. schmitti wild shrimp and in the C. danae blue crab is reported for the first time in the present work. The results indicate that the main species of shrimps and blue crabs of the environment surrounding the farms were infected by WSSV, and they may be considered potential vectors of the virus.

  6. Presença do vírus da síndrome da mancha branca em crustáceos decápodes silvestres em lagoas costeiras no Sul do Brasil Presence of the white spot syndrome virus (WSSV in wild decapods crustaceans in coastal lagoons in southern Brazil

    Directory of Open Access Journals (Sweden)

    S.W. Costa

    2012-02-01

    Full Text Available A presença do vírus da síndrome da mancha branca (em inglês WSSV nas principais espécies de camarões, siris e caranguejos de cinco lagoas que recebem o efluente de fazendas afetadas pela enfermidade foi detectada por nested PCR, e inclusões virais nos camarões por histologia. Pela nested PCR encontrou-se a presença de WSSV em 13 de 16 (81,2% amostras de camarões da espécie Farfantepenaeus paulensis, em 13 de 14 (92,8% de Litopenaeus schmitti, em uma de duas de Farfantepenaeus brasiliensis (50%, em 13 de 15 (86,6% de siri da espécie Callinectes danae e em 11 de 12 (91,6% de Callinectes sapidus, e não foi detectada no caranguejo Chasmagnathus granulata em 10 amostras. Inclusões características de WSSV foram observadas em três amostras histológicas de 50 (6,0% no epitélio gástrico e cuticular e nas brânquias de dois exemplares de F. paulensis e um de L. schmitti. É o primeiro relato da presença de WSSV em camarões L. schmitti e no siri C. danae silvestres. As principais espécies de camarões e siris dos ambientes de entorno das fazendas foram contaminadas pelo WSSV, constituindo-se em vetores potenciais do vírus.The presence of white spot syndrome virus (WSSV in the main species of shrimps, blue crabs, and burrowing crabs of five lagoons where shrimp farm effluents are discharged, was analyzed by nested PCR and the presence of virus inclusions in the shrimps was analyzed through histopathology. The nested PCR analysis indicated the presence of WSSV in 13 of 16 (81.2% samples of the shrimp species of Farfantepenaeus paulensis, in 13 of 14 (92.8% of Litopenaeus schmitti, in one of two of Farfantepenaeus brasiliensis (50%, in 13 of 15 (86.6% of blue crab species of Callinectes danae and in 11 of 12 (91.6% of Callinectes sapidus and none was detected in the 10 samples of the burrowing crab Chasmagnathus granulata. The inclusion characteristics of WSSV were observed in three samples of 50 (6.0% in the gastric and cuticular epithelium and in the gills of two specimens of F. paulensis and one of L. schmitti. The presence of WSSV in L. schmitti wild shrimp and in the C. danae blue crab is reported for the first time in the present work. The results indicate that the main species of shrimps and blue crabs of the environment surrounding the farms were infected by WSSV, and they may be considered potential vectors of the virus.

  7. The shrimp IKK–NF-?B signaling pathway regulates antimicrobial peptide expression and may be subverted by white spot syndrome virus to facilitate viral gene expression

    OpenAIRE

    Wang, Pei-Hui; Gu, Zhi-Hua; Wan, Ding-Hui; Liu, Bo-Du; Huang, Xian-De; Weng, Shao-ping; Yu, Xiao-Qiang; He, Jian-Guo

    2013-01-01

    The I?B kinases IKK? and IKK? and the IKK-related kinases TANK-binding kinase 1 (TBK1) and IKK? are the master regulators of the NF-?B signaling pathway. Although this pathway has been extensively studied in mammals, less attention has been paid in crustaceans, which have significant economic value. Here, we report the cloning and functional studies of two IKK homologs, LvIKK? and LvIKK?, from Pacific white shrimp, Litopenaeus vannamei. LvIKK? and LvIKK? mRNAs are widely expressed in...

  8. Risk factors and management of white spot lesions in orthodontics

    OpenAIRE

    Srivastava, Kamna; Tikku, Tripti; Khanna, Rohit; Sachan, Kiran

    2013-01-01

    The formation of white spot lesions or enamel demineralization around fixed orthodontic attachments is a common complication during and following fixed orthodontic treatment, which mars the result of a successfully completed case. This article is a contemporary review of the risk factors, preventive methods and fate of these orthodontics scars. The importance of excellent oral hygiene practice during fixed orthodontic treatment must be explained. Preventive programs must be emphasized to all ...

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

    OpenAIRE

    Oliveira Resende, R., de

    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, 1971). Under experimental conditions however, it is common practice to maintain the virus by mechanical inoculation onto susceptible host plants.Repeated passages of animal viruses, certainly at high inoculum ...

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

    OpenAIRE

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

  11. Status of tobacco viruses in Serbia and molecular characterization of tomato spotted wilt virus isolates.

    Science.gov (United States)

    Stankovi?, I; Bulaji?, A; Vu?urovi?, A; Risti?, D; Milojevi?, K; Berenji, J; Krsti?, B

    2011-01-01

    In a four-year survey to determine the presence and distribution of viruses in tobacco crops at 17 localities of the Vojvodina Province and Central Serbia, 380 samples were collected and analyzed by DAS-ELISA. Out of the seven viruses tested, tomato spotted wilt virus (TSWV), potato virus Y (PVY), tobacco mosaic virus (TMV), cucumber mosaic virus (CMV), and alfalfa mosaic virus (AMV) were detected in 37.9, 33.4, 28.7, 23.9, and 15.5% of the total tested samples, respectively. TSWV was the most frequently found virus at the localities of Central Serbia, while PVY and CMV were the most frequent viruses in the Vojvodina Province. Single infections were prevalent in years 2005-2007 and the most frequent were those of PVY. A triple combination of those viruses was most frequent mixed infection type in 2008. The presence of all five detected viruses was confirmed in selected ELISA-positive samples by RT-PCR and sequencing. The comparisons of obtained virus isolate sequences with those available in NCBI, confirmed the authenticity of serologically detected viruses. Phylogenetic analysis based on partial nucleocapsid gene sequences revealed a joint clustering of Serbian, Bulgarian and Montenegrin TSWV isolates into one geographic subpopulation, which was distinct from the other subpopulation of TSWV isolates from the rest of the European countries. The high incidence of viruses in Serbian tobacco crops highlights the importance of enhancing farmers knowledge towards better implementation of control strategies for preventing serious losses. PMID:22149499

  12. Effects of Smart Position Only (SPOT) Tag Deployment on White Sharks Carcharodon carcharias in South Africa

    OpenAIRE

    Jewell, Oliver Joseph David; Wcisel, Michelle A.; Gennari, Enrico; Towner, Alison V.; Bester, Marthan Nieuwoudt; Johnson, Ryan Lloyd; Singh, Sarika

    2011-01-01

    We present 15 individual cases of sub-adult white sharks that were SPOT tagged in South Africa from 2003–2004 and have been re-sighted as recently as 2011. Our observations suggest SPOT tags can cause permanent cosmetic and structural damage to white shark dorsal fins depending on the duration of tag attachment. SPOT tags that detached within 12–24 months did not cause long term damage to the dorsal fin other than pigmentation scarring. Within 12 months of deployment, tag foul...

  13. The Shrimp NF-?B Pathway Is Activated by White Spot Syndrome Virus (WSSV) 449 to Facilitate the Expression of WSSV069 (ie1), WSSV303 and WSSV371

    OpenAIRE

    Wang, Pei-Hui; Gu, Zhi-Hua; Wan, Ding-Hui; Zhang, Ming-Yan; Weng, Shao-ping; Yu, Xiao-Qiang; He, Jian-guo

    2011-01-01

    The Toll-like receptor (TLR)-mediated NF-?B pathway is essential for defending against viruses in insects and mammals. Viruses also develop strategies to utilize this pathway to benefit their infection and replication in mammal hosts. In invertebrates, the TLR-mediated NF-?B pathway has only been well-studied in insects and has been demonstrated to be important in antiviral responses. However, there are few reports of interactions between viruses and the TLR-mediated NF-?B pathway in inverteb...

  14. White Matter Damage in Brain May Help Spot Early Alzheimer's

    Science.gov (United States)

    ... Matter Damage in Brain May Help Spot Early Alzheimer's Study finds widespread changes using specialized MRI To ... be an early sign of certain types of Alzheimer's disease, according to a new study. Researchers used ...

  15. ????????????????? The Forming Mechanism of the White Spot on the Chemical Conversion Coating of the Die-Casting Magnesium Alloy

    Directory of Open Access Journals (Sweden)

    ???

    2013-03-01

    Full Text Available ???????????????????????????????????SEM?????????????????????????????????????????????????????????????????????????????????????????????????????????????The forming mechanism of the white spot of the die-casting magnesium was investigated by means of SEM andOM.The roughness of the white spot was also tested. It indicated that the larger ? phase caused by the uneven temperature distribution of the die result in the formation of the white spot. It can be improved by changing the agent spaying parameter.

  16. Two variants in the KIT gene as candidate causative mutations for a dominant white and a white spotting phenotype in the donkey.

    Science.gov (United States)

    Haase, B; Rieder, S; Leeb, T

    2015-06-01

    White spotting phenotypes have been intensively studied in horses, and although similar phenotypes occur in the donkey, little is known about the molecular genetics underlying these patterns in donkeys. White spotting in donkeys can range from only a few white areas to almost complete depigmentation and is characterised by a loss of pigmentation usually progressing from a white spot in the hip area. Completely white-born donkeys are rare, and the phenotype is characterised by the complete absence of pigment resulting in pink skin and a white coat. A dominant mode of inheritance has been demonstrated for spotting in donkeys. Although the mode of inheritance for the completely white phenotype in donkeys is not clear, the phenotype shows similarities to dominant white in horses. As variants in the KIT gene are known to cause a range of white phenotypes in the horse, we investigated the KIT gene as a potential candidate gene for two phenotypes in the donkey, white spotting and white. A mutation analysis of all 21 KIT exons identified a missense variant in exon 4 (c.662A>C; p.Tyr221Ser) present only in a white-born donkey. A second variant affecting a splice donor site (c.1978+2T>A) was found exclusively in donkeys with white spotting. Both variants were absent in 24 solid-coloured controls. To the authors' knowledge, this is the first study investigating genetic mechanisms underlying white phenotypes in donkeys. Our results suggest that two independent KIT alleles are probably responsible for white spotting and white in donkeys. PMID:25818843

  17. Toll-pathway in tiger shrimp (Penaeus monodon) responds to white spot syndrome virus infection: evidence through molecular characterisation and expression profiles of MyD88, TRAF6 and TLR genes.

    Science.gov (United States)

    Deepika, A; Sreedharan, K; Paria, Anutosh; Makesh, M; Rajendran, K V

    2014-12-01

    The Toll-pathway plays key roles in regulating the innate immune response in invertebrates. Myeloid differentiation factor 88 (MyD88) and Tumour necrosis factor receptor (TNFR)-associated factor 6 (TRAF6) are key molecules in this signalling pathway. To investigate the role of Toll-pathway in innate immune response of shrimp, Penaeus monodon, MyD88 (PmMyD88) and TRAF6 (PmTRAF6) were identified and characterised. PmMyD88 cDNA is 1716 bp long with an open reading frame (ORF) of 1449 bp encoding a putative protein of 482 amino acids, with a death domain, a TIR domain and C-terminal extension domain. PmTRAF6 cDNA is 2563 bp long with an ORF of 1785 bp (594 amino acids) with an N-terminal RING-type zinc finger domain, two TRAF-type zinc finger domains, a coiled region and a MATH domain. In healthy shrimp, PmMyD88, PmTRAF6 and PmToll were detected in 15 tissues with the highest expression in midgut, eyestalk and lymphoid organ, respectively. Responses of these genes to WSSV in experimentally-infected P. monodon as well as in cultured haemocytes and also effect of poly I:C on the gene expression in vitro was investigated at six time-points in seven tissues. PmToll showed significant up-regulation at all time-points of infection in six tissues and until 24 h post-infection in vitro. However, poly I:C-induced haemocytes showed up-regulation of the gene until 48 h post-exposure. WSSV caused significant up-regulation of PmMyD88 in most of the tissues tested. The virus challenge as well as poly I:C induction in vitro also resulted in significant up-regulation of the gene. Up-regulated expression of PmTRAF6 was detected in haemocytes and lymphoid organ at late stage of infection. In vitro virus challenge showed significant up-regulation of PmTRAF6 at almost all time-points whereas no significant change in the expression was observed on poly I:C induction. The responses of these key genes, observed in the present study, suggest that Toll-pathway as a whole may play a crucial role in the immune response against viruses in shrimp. PMID:25266891

  18. Bacillus subtilis spores expressing the VP28 antigen: a potential oral treatment to protect Litopenaeus vannamei against white spot syndrome.

    Science.gov (United States)

    Nguyen, Anh T V; Pham, Cuong K; Pham, Huong T T; Pham, Hang L; Nguyen, Anh H; Dang, Lua T; Huynh, Hong A; Cutting, Simon M; Phan, Tuan-Nghia

    2014-09-01

    The envelope protein VP28 of white spot syndrome virus (WSSV) is considered a candidate antigen for use in a potential vaccine to this important shrimp pathogen (the cause of white spot syndrome, WSS). Here, we used spores of Bacillus subtilis to display VP28 on the spore surface. Trials were conducted to evaluate their ability to protect shrimps against WSSV infection. The gene cotB-vp28 was integrated into the chromosome of the laboratory strain B. subtilis PY79, and expression of CotB-VP28 was detected by Western blotting and immunofluorescence. Expression of CotB-VP28 was equivalent to 1000 molecules per spore. PY79 and CotB-VP28 spores were mixed with pellets for feeding of whiteleg shrimps (Litopenaeus vannamei), followed by WSSV challenge. Superoxidase dismutase (SOD), phenoloxidase activities and mortality rates of the two shrimp groups were evaluated. Groups fed with PY79 and CotB-VP28 spores at day 7 had increased SOD activities of 29% and increased phenoloxidase activities of 15% and 33%, respectively, compared to those of the control group. Fourteen days postchallenge, 35% of vaccinated shrimps had died compared to 49% of those fed naked spores (PY79) and 66% untreated, unchallenged animals. These data suggest that spores expressing VP28 have potential as a prophylactic treatment of WSS. PMID:25066511

  19. Genotyping WSSV isolates from northwestern Mexican shrimp farms affected by white spot disease outbreaks in 2010-2012.

    Science.gov (United States)

    Durán-Avelar, Ma de; Pérez-Enríquez, Ricardo; Zambrano-Zaragoza, José Francisco; Montoya-Rodríguez, Leobardo; Vázquez-Juárez, Ricardo; Vibanco-Pérez, Norberto

    2015-05-11

    White spot disease (WSD) causes high mortality in cultured shrimp throughout the world. Its etiologic agent is the white spot syndrome virus (WSSV). The genomic repeat regions ORF 75, ORF 94, and ORF 125 have been used to classify WSSV isolates in epidemiological studies using PCR with specific primers and sequencing. The present study investigated the variation in nucleotide sequences from 107, 150, and 143 isolates of WSSV collected from Litopenaeus vannamei shrimp ponds with WSD outbreaks in northwestern Mexico during the period 2010-2012, in the genomic repeat regions ORFs 75, 94, and 125, respectively. The haplotypic nomenclature for each isolate was based on the number of repeat units and the position of single nucleotide polymorphisms on each ORF. We report finding 17, 43, and 66 haplotypes of ORFs 75, 94, and 125, respectively. The study found high haplotypic diversity in WSSV using the complete sequences of ORFs 94 and 125 as independent variables, but low haplotypic diversity for ORF 75. Different haplotypes of WSSV were found from region-to-region and year-to-year, though some individual haplotypes were found in different places and in more than one growing cycle. While these results suggest a high rate of mutation of the viral genome at these loci, or perhaps the introduction of new viral strains into the area, they are useful as a tool for epidemiological surveys. Two haplotypes from some of the ORFs in the same shrimp were encountered, suggesting the possibility of multiple infections. PMID:25958803

  20. 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

    Scientific Electronic Library Online (English)

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

    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, microsco [...] pia 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. Abstract in english 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.

  1. 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.

  2. 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. PMID:25904110

  3. Sequence analysis of Indian iris yellow spot virus ambisense genome segments: evidence of interspecies RNA recombination.

    Science.gov (United States)

    Gawande, S J; Gurav, V S; Ingle, A A; Martin, D P; Asokan, R; Gopal, J

    2015-05-01

    The nucleotide sequence of M- and S-RNA segments of an Indian iris yellow spot virus (IYSV) were determined. Sequence comparisons showed that both of these sequences shared less than 95 % identity with those other known IYSV isolates. Phylogenetic analysis revealed that the S- and M-RNA sequences of known IYSV isolates clustered with those of the tospoviruses, tomato yellow ring virus, polygonum ringspot virus and hippeastrum chlorotic ringspot virus. Further, multiple recombination detection methods detected inter- and intra-species recombination events that clustered primarily within the intergenic regions of S- and M-RNA, suggesting that these are possibly recombination hotspots in IYSV and closely related tospoviruses. PMID:25655262

  4. Endogenous Retrovirus Insertion in the KIT Oncogene Determines White and White spotting in Domestic Cats

    OpenAIRE

    David, Victor A.; Menotti-raymond, Marilyn; Wallace, Andrea Coots; Roelke, Melody; Kehler, James; Leighty, Robert; Eizirik, Eduardo; Hannah, Steven S.; Nelson, George; Scha?ffer, Alejandro A.; Connelly, Catherine J.; O’brien, Stephen J.; Ryugo, David K.

    2014-01-01

    The Dominant White locus (W) in the domestic cat demonstrates pleiotropic effects exhibiting complete penetrance for absence of coat pigmentation and incomplete penetrance for deafness and iris hypopigmentation. We performed linkage analysis using a pedigree segregating White to identify KIT (Chr. B1) as the feline W locus. Segregation and sequence analysis of the KIT gene in two pedigrees (P1 and P2) revealed the remarkable retrotransposition and evolution of a feline endogenous retrovirus (...

  5. Analysis of the transcription initiation mechanism of tomato spotted wilt virus

    OpenAIRE

    Duijsings, D.M.J.M.

    2001-01-01

    Genome replication and transcription of Tomato spotted wilt virus (TSWV, genus Tospovirus ) follows in most aspects the general rules for negative strand RNA viruses with segmented genomes. One common feature is the occurrence of "cap snatching" during transcription initiation. During this process, capped leader sequences of suitable host mRNAs are recruited and cleaved to serve as primers for transcription.At the start of the research project described in this thesis, the occurrence of cap s...

  6. 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.

  7. FIRST REPORT OF TOMATO SPOTTED WILT VIRUS IN BLACKBERRY LILY IN NORTH AMERICA

    Science.gov (United States)

    Blackberry lily [Belamcanda chinensis (L.) DC.] is an herbaceous perennial in the Iridaceae characterized by purple-spotted orange flowers followed by persistent clusters of black fruit. We observed virus-like symptoms including chlorotic ringspots and ring patterns on blackberry lily leaves in nor...

  8. Enhancement of superoxide dismutase and catalase activity in juvenile brown shrimp, Farfantepenaeus californiensis (Holmes, 1900, fed ?-1.3 glucan vitamin E, and ?-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 ?-1,3 glucano vitamina E y ?-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 ?-?-glucan, vitamin E, and ?-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 ?-1.3-glucan (0.1%, vitamin E (0.01%, and ?-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 ?-1.3-glucan, vitamin E, and ?-carotene significantly increased SOD activity in the hepatopancreas and muscle at 12 and 24 h post-infection, respectively. Shrimp fed with vitamin E and ?-1.3-glucan registered an increment in SOD activity from 12 to 48 h post-infection. Shrimp fed with ?-carotene increased SOD activity before infection with WSSV, and shrimp fed with ?-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 ?-?-glucan and ?-carotene showed greater resistance to WSSV, with mortality at 144 h post-infection. This study demonstrated the capacity of juvenile Farfantepenaeus californiensis fed ?-?-glucan, vitamin E, or ?-carotene to increase the antioxidant response before and after viral infection.Se evaluó el efecto de ?-1,3-glucano, vitamina E y ?-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 contenedores de fibra de vidrio de 60 L y se alimentaron diariamente durante 23 días con ?-1,3-glucano (0,1%, vitamina E (0,01%, y ?-caroteno (0,01% y posteriormente se inocularon con WSSV. Se determinó la actividad antioxidante de la enzima superóxido dismutasa (SOD y catalasa (CAT en hepatopáncreas y músculo a las 0, 1, 6, 12, 24 y 48 h después de la infección. Los grupos de camarones alimentados con los tratamientos incrementaron la actividad SOD en el hepatopáncreas y músculo a las 12 y 24 h después de la infección, respectivamente. Los juveniles tratados con vitamina E y ?-1,3-glucano mantuvieron un incremento en la actividad SOD desde las 12 a 48 h postinfección. Los camarones alimentados con ?-caroteno incrementaron la actividad de SOD antes de la infección con WSSV y los que fueron alimentados con ?-1,3-glucano y vitamina E incrementaron la actividad CAT también antes de la infección. La actividad CAT en músculo se incrementó respecto al grupo control, con todos los grupos de camarones tratados desde 1 hasta 6 h posteriores a la inoculación con WSSV. La actividad antioxidante más alta se registró en los camarones alimentados con vitamina E. Los juveniles alimentados con vitamina E y posteriormente inoculados con WSSV, registraron 100% de mortalidad a las 72 h, pero los que fueron alimentados con ?-1,3-glucano y ?-caroteno resistieron la infección hasta las 144 h. Los resultados de Antioxidant response in F. californiensis fed with dietary supplements and infected with WSSV este estudio mostraron la capacidad de juveniles de Farfantepenaeus californiensis alimentados con ?-1,3-glucano, vitamina E o ?-caroteno, de incrementar la respuesta antioxidante antes y durante una infección viral.

  9. Enhancement of superoxide dismutase and catalase activity in juvenile brown shrimp, Farfantepenaeus californiensis (Holmes, 1900), fed ?-1.3 glucan vitamin E, and ?-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 ?-1,3 glucano vitamina E y ?-caroteno e infectados con el virus de la mancha blanca

    Scientific Electronic Library Online (English)

    Rosario, Pacheco; Felipe, Ascencio; Martha, Zarain; Gracia, Gómez; Ángel, Campa.

    2011-11-01

    Full Text Available Se evaluó el efecto de ?-1,3-glucano, vitamina E y ?-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 contenedores de fibra de vidrio de 60 L y se ali [...] mentaron diariamente durante 23 días con ?-1,3-glucano (0,1%), vitamina E (0,01%), y ?-caroteno (0,01%) y posteriormente se inocularon con WSSV. Se determinó la actividad antioxidante de la enzima superóxido dismutasa (SOD) y catalasa (CAT) en hepatopáncreas y músculo a las 0, 1, 6, 12, 24 y 48 h después de la infección. Los grupos de camarones alimentados con los tratamientos incrementaron la actividad SOD en el hepatopáncreas y músculo a las 12 y 24 h después de la infección, respectivamente. Los juveniles tratados con vitamina E y ?-1,3-glucano mantuvieron un incremento en la actividad SOD desde las 12 a 48 h postinfección. Los camarones alimentados con ?-caroteno incrementaron la actividad de SOD antes de la infección con WSSV y los que fueron alimentados con ?-1,3-glucano y vitamina E incrementaron la actividad CAT también antes de la infección. La actividad CAT en músculo se incrementó respecto al grupo control, con todos los grupos de camarones tratados desde 1 hasta 6 h posteriores a la inoculación con WSSV. La actividad antioxidante más alta se registró en los camarones alimentados con vitamina E. Los juveniles alimentados con vitamina E y posteriormente inoculados con WSSV, registraron 100% de mortalidad a las 72 h, pero los que fueron alimentados con ?-1,3-glucano y ?-caroteno resistieron la infección hasta las 144 h. Los resultados de Antioxidant response in F. californiensis fed with dietary supplements and infected with WSSV este estudio mostraron la capacidad de juveniles de Farfantepenaeus californiensis alimentados con ?-1,3-glucano, vitamina E o ?-caroteno, de incrementar la respuesta antioxidante antes y durante una infección viral. Abstract in english The effect of dietary ?-?-glucan, vitamin E, and ?-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 ?-1. [...] 3-glucan (0.1%), vitamin E (0.01%), and ?-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 ?-1.3-glucan, vitamin E, and ?-carotene significantly increased SOD activity in the hepatopancreas and muscle at 12 and 24 h post-infection, respectively. Shrimp fed with vitamin E and ?-1.3-glucan registered an increment in SOD activity from 12 to 48 h post-infection. Shrimp fed with ?-carotene increased SOD activity before infection with WSSV, and shrimp fed with ?-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 ?-?-glucan and ?-carotene showed greater resistance to WSSV, with mortality at 144 h post-infection. This study demonstrated the capacity of juvenile Farfantepenaeus californiensis fed ?-?-glucan, vitamin E, or ?-carotene to increase the antioxidant response before and after viral infection.

  10. 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...

  11. Effects of Microabrasion on non Active White Spots in Primary Teeth

    Directory of Open Access Journals (Sweden)

    Carolina Júdica RAMOS

    2006-08-01

    Full Text Available Objective: The aim of this study was to verify the enamel micromorphology with non active white spot lesions after the application of two different compounds used in microabrasion technique. Method: Thirty primary molars with non active white spot lesions in approximal surfaces were selected. Samples were randomly divided into three groups. G1 (n=10: control, untreated. G2 (n=10: compound (35% phosphoric acid and pumice mixed - 1:1 was applied on stain enamel. G3 (n=10: compound Opalustre, a slurry containing 6.6% HCl (hydrochloric acid and silicon carbide microparticles was applied. After this, all specimens were dehydrated, mounted on metal stubs, coated with gold and examined under scanning electron microscope (Jeol, JSM - 6100. Results: Eletronmicrographs analysis showed that the G1 samples (control, without treatment had a smooth surface with the presence of pumice residue. G2 samples (phosphoric acid and pumice presented rugous enamel surface with areas of enamel prisms demineralization. In G3 samples (Opalustre it were observed the formation of scratchs and grooves, beyond the presence of compound residue. Conclusion: The application of different types of compounds led to the formation of different aspects in micromorphology of primary teeth enamel.

  12. 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 by mechanical infection and by the vector F. accidentalis. Moreover, evolved strains, here indicated as TSWV-RB-PI (Pepper, Italy), are not able to infect tomato plants carrying the Sw5 gene, confirming that distinct viral gene products control the outcome of infection in plants carrying Sw5 and Tsw. The simultaneous presence of evolved TSWV strains, showing the same biological characteristics (such as host range and symptomatology on different hosts) in two geographic areas of southern Italy, very far each other, let us hypothesize a unique focus of infection. Only subsequently, infected plants, through trade of plant material, have been diffused in locations so far apart. Further molecular studies are underway to assess the possible correlation between the different isolates in different geographical areas. PMID:25151838

  13. A spot-PCR technique for the detection of phloem-limited grapevine viruses.

    Science.gov (United States)

    La Notte, P; Minafra, A; Saldarelli, P

    1997-06-01

    Specific amplification of genomic fragments of grapevine trichovirus A (GVA), grapevine trichovirus B (GVB) and grapevine leafroll-associated closterovirus 3 (GLRaV3) was obtained by reverse transcription-PCR on total nucleic acid solubilized from small pieces of charged nylon membrane, on which a drop of crude infected grapevine sap was spotted (spot-PCR). A thermal treatment (95 degrees for 10 degrees) of spotted sap in a buffered solution improved the release of viral template. Consistent amplification was obtained with three viruses from fragments of the same respective blots up to 1 month after spotting, while a detection threshold limit comparable with standard PCR techniques was found for this method. Duplex PCR (i.e. amplification of different viruses from a mixed-infected grapevine source) was also found to be effective, since GVA and GLRaV3 were amplified by a mixture of specific primers in the same reaction. This rapid and easy sampling technique, using leaf petioles to express crude sap, may have a wide field application for screening grapevine viruses. PMID:9220395

  14. Controle químico da Cercosporiose, Mancha-Branca e dos Grãos Ardidos em milho / Chemical control of gray leaf spot, white spot and rot grain in maize

    Scientific Electronic Library Online (English)

    André Humberto, Brito; Renzo Garcia, Von Pinho; José Luiz de Andrade Rezende, Pereira; Marcio, Balestre.

    2013-10-01

    Full Text Available Este trabalho foi realizado com o objetivo de avaliar os efeitos da aplicação de fungicida, no controle da Cercosporiose e da Mancha-Branca do milho, e suas consequências na produtividade de grãos e na incidência de grãos ardidos, além de estabelecer a relação existente entre a produtividade de grão [...] s e a severidade dessas doenças. Dois experimentos distintos (com Azoxystrobina + Cyproconazole e tratamento controle) foram conduzidos em três locais, no ano agrícola de 2007/2008. Utilizaram-se 12 híbridos comerciais de milho, que foram avaliados em delineamento de blocos casualizados, com três repetições. Foram realizadas cinco avaliações da severidade das doenças Cercosporiose e Mancha-Branca, por meio de escala de notas, variando de 1 (altamente resistente) a 9 (altamente susceptível). Estimou-se a área abaixo da curva de progresso da doença (AACPD). A aplicação de fungicida é eficiente no controle de doenças foliares e na redução da incidência de grãos ardidos, além de proporcionar 12% de aumento na produtividade de grãos, em relação à do tratamento controle. As doenças foliares Cercosporiose e Mancha-Branca reduzem a produtividade de grãos de milho e essa redução é maior quando as doenças ocorrem mais precocemente. A Cercosporiose provoca maior redução na produtividade de grãos, quando comparada com a Mancha-Branca. Abstract in english This work was carried out in order to evaluate the effects of fungicides on the control of maize gray leaf spot and maize white spot and its implications on the grain yield and rot grain, as well as to establish the relationship between grain yield and disease severity. Twelve commercial single cros [...] s hybrids were evaluated in a randomized block design with three repetitions. Five evaluations of disease severity (maize white spot and maize gray leaf spot) were performed using scale-visual symptoms, with scores ranging from 1 (highly resistant) to 9 (highly susceptible). In addition, the area under the disease progress curve (AUDPC) was estimated. Fungicide application is effective in the control of maize leaf diseases, and the experiments without control of the diseases had grain yield lower than that of the experiments with control, with a reduction of 1.2 t.ha-1 (12.3%) on average. The fungicide application had a positive effect in the control of rot grains, with a reduction of 2.6% on average. Maize gray leaf spot and maize white spot reduce the maize grain yield and this reduction is greater when diseases occur earlier. Gray leaf spot causes a greater reduction in grain yield compared with white spot.

  15. White spots on Smoke rings by Bruce Nauman: a case study on contemporary art conservation using microanalytical techniques.

    Science.gov (United States)

    Mafalda, Ana Cardeira; da Câmara, Rodrigo Bettencourt; Strzelec, Patrick; Schiavon, Nick; Mirão, José; Candeias, António; Carvalho, Maria Luísa; Manso, Marta

    2015-02-01

    The artwork "Smoke Rings: Two Concentric Tunnels, Non-Communicating" by Bruce Nauman represents a case study of corrosion of a black patina-coated Al-alloy contemporary artwork. The main concern over this artwork was the widespread presence of white spots on its surface. Alloy substrate, patina, and white spots were characterized by means of energy-dispersive X-ray fluorescence and scanning electron microscopy with energy-dispersive spectroscopy. Alloy substrate was identified as an aluminum alloy 6,000 series Al-Si-Mg. Patina's identified composition confirmed the documentation provided by the atelier. Concerning the white spots, zircon particles were found on patina surface as external elements. PMID:25158782

  16. Tomato chlorotic spot virus in hydroponically-grown lettuce in São Paulo State, Brazil

    Directory of Open Access Journals (Sweden)

    Colariccio Addolorata

    2004-01-01

    Full Text Available In the regions of Campinas and Sumaré, São Paulo, Brazil, hidroponically grown crops of Lettuce (Lactuca sativa cv. Verônica, which showed virus-like symptoms were examined by electron microscope, biological, serological and molecular tests. Pleomorphic, enveloped particles (80-100 nm in diameter were always detected in these samples. Experimentally inoculated host plants, including lettuce, reacted with tospoviruses-induced symptoms. Some differences were observed in Gomphrena globosa, which reacted by showing local lesions and systemic mosaic. Two isolates of Tomato chlorotic spot virus (TCSV were identified by DAS-ELISA and by RT-PCR. The sequencing and alignment of the RT-PCR coat protein amplified fragments have indicated a high degree of homology with the TCSV sequences stored in the GenBank. This is the first report of losses due to a virus from the genus Tospovirus in commercial hydroponic lettuce crops in Brazil. Further epidemiological studies are needed for better understanding the spread of the virus in hydroponic crops, since Tomato spotted wilt virus (TSWV is reported to spread through the nutritive solution.

  17. Identifying geographic hot spots of reassortment in a multipartite plant virus

    OpenAIRE

    Savory, Fiona R.; Varma, Varun; Ramakrishnan, Uma

    2014-01-01

    Reassortment between different species or strains plays a key role in the evolution of multipartite plant viruses and can have important epidemiological implications. Identifying geographic locations where reassortant lineages are most likely to emerge could be a valuable strategy for informing disease management and surveillance efforts. We developed a predictive framework to identify potential geographic hot spots of reassortment based upon spatially explicit analyses of genome constellatio...

  18. HEPATITIS B VIRUS DNA CAN BE AMPLIFIED DIRECTLY FROM DRIED BLOOD SPOT ON FILTER PAPER

    OpenAIRE

    Naglaa Fathy Alhusseini; Abadeer, Mamdouh Z.; El-taher, Sheref M.

    2012-01-01

    Hepatitis B Virus (HBV) infections represent a major public health problem because of the ability of HBV to cause a chronic carrier state. Even though chronic carriers remain largely asymptomatic, a large number of these individuals subsequently develop cirrhosis and primary hepatocellular carcinomas. Dried Blood Spot (DBS) samples are a simple and inexpensive sampling method, especially useful for blood collection in resource poor settings with limited access to diagnostic facilities. The ma...

  19. Qualitative Human Immunodeficiency Virus RNA Analysis of Dried Blood Spots for Diagnosis of Infections in Infants?

    OpenAIRE

    Kerr, Ryan J. S.; Player, Gray; Fiscus, Susan A.; Nelson, Julie A. E.

    2008-01-01

    The Gen-Probe Aptima human immunodeficiency virus type 1 (HIV-1) RNA assay was adapted for the diagnosis of HIV infection in infants by using dried blood spots. The assay was 99% sensitive (128/129) and 100% specific (162/162). This may prove useful in resource-limited settings, since it precludes the need for a phlebotomist and maintenance of a cold chain.

  20. Prevalence of enamel white spots and risk factors in children up to 36 months old

    Scientific Electronic Library Online (English)

    Ana Valéria Pagliari, Tiano; Suzely Adas Saliba, Moimaz; Orlando, Saliba; Cléa Adas Saliba, Garbin.

    2009-06-01

    Full Text Available The aim of this study was to determine the prevalence of enamel white spots and the quality of oral hygiene in children up to 36 months old, in municipalities with different fluoride levels in the water supply, analyzing the contribution of several variables. After approval of the Ethics Committee, [...] the parents signed an informed consent form and were interviewed about their educational level, economic classification of the family, nursing habits, use of toothpaste, access to dental service and other information. The children were clinically examined using the same codes and criteria established by the WHO (World Health Organization) and ADA (American Dental Association). The data were processed and analyzed with the Epi-info software program, version 3.2, and Microsoft Excel. Fisher's exact test (p

  1. Treatment of post-orthodontic white spot lesions with casein phosphopeptide-stabilised amorphous calcium phosphate

    DEFF Research Database (Denmark)

    Bröchner, Ann; Christensen, Carsten

    2011-01-01

    This study aims to investigate the effect of topical applications of 10% casein phosphopeptide-amorphous calcium phosphate (CPP-ACP) on white spot lesions (WSL) detected after treatment with fixed orthodontic appliances. Sixty healthy adolescents with >/=1 clinically visible WSL at debonding were recruited and randomly allocated to a randomised controlled trial with two parallel groups. The intervention group was instructed to topically apply a CPP-ACP -containing agent (Tooth Mousse, GC Europe) once daily and the subjects of the control group brushed their teeth with standard fluoride toothpaste. The intervention period was 4 weeks and the endpoints were quantitative light-induced fluorescence (QLF) on buccal surfaces of the upper incisors, cuspids and first premolars and visual scoring from digital photos. The attrition rate was 15%, mostly due to technical errors, and 327 lesions were included in the final evaluation. A statistically significant (p <0.05) regression of the WSL was disclosed in both study groups compared to baseline, but there was no difference between the groups. The mean area of the lesions decreased by 58% in the CPP-ACP group and 26% in the fluoride group (p = 0.06). The QLF findings were largely reflected by the clinical scores. No side effects were reported. Topical treatment of white spot lesions after debonding of orthodontic appliances with a casein phosphopeptide-stabilised amorphous calcium phosphate agent resulted in significantly reduced fluorescence and a reduced area of the lesions after 4 weeks as assessed by QLF. The improvement was however not superior to the "natural" regression following daily use of fluoride toothpaste.

  2. Fontes de resistência em tomateiro aos begomovírus bissegmentados Tomato yellow spot virus e Tomato severe rugose virus Sources of resistance in tomato to bipartite begomoviruses Tomato yellow spot virus and Tomato severe rugose virus

    Directory of Open Access Journals (Sweden)

    Francisco D Hurtado

    2012-12-01

    Full Text Available Os begomovírus causam doenças de grande importância econômica em diversas culturas, principalmente em regiões tropicais e subtropicais. Juntamente com outras famílias de vírus, os begomovírus têm causado grande prejuízo para os produtores de tomate in natura e para processamento industrial. O objetivo deste trabalho foi avaliar o comportamento de 11 genótipos resistentes ao Tomato yellow leaf curl virus (TYLCV frente à infecção pelos begomovírus Tomato yellow spot virus (ToYSV e Tomato severe rugose virus (ToSRV em condições de casa-vegetação. A inoculação das plantas foi realizada via biobalística no estádio de duas folhas verdadeiras. A infecção viral confirmou-se pelo desenvolvimento dos sintomas e pela técnica de hibridização dot blot. Selecionaram-se como promissores os genótipos STY2, STY5, STY6 e L7, por não apresentarem sintomas e por terem concentrações virais muito baixas para os dois vírus. O espectro de resistência dos genes Ty-1 e Ty-2 não resultaram efetivos ante as espécies virais empregadas no estudo. As linhagens TY52, H24 e CLN2116B, portadoras destes genes, foram suscetíveis aos vírus ToYSV e ToSRV.Begomoviruses cause diseases of major economic importance in many crops, especially in tropical and subtropical regions. Together with other families of viruses, the begomoviruses cause great damage for producers of fresh and processed tomatoes. The objective of this study was to evaluate the behavior of 11 resistant genotypes to Tomato yellow leaf curl virus (TYLCV when infected by the begomoviruses Tomato severe rugose virus (ToSRV and Tomato yellow spot virus (ToYSV under greenhouse conditions. Tomato plants were inoculated by biobalistic method when displaying two true leaves. Viral infection was confirmed by visual observation of symptoms and by dot blot hybridization. Genotypes STY2, STY5, STY6 and L7 were selected as promising due to the absence of symptoms and very low viral concentration for both viruses. The resistance genes Ty-1 and Ty-2 were not effective against the viral species used in the study. The lines TY52, H24 and CLN2116B, carrying these genes, exhibited a susceptible behavior.

  3. Fontes de resistência em tomateiro aos begomovírus bissegmentados Tomato yellow spot virus e Tomato severe rugose virus / Sources of resistance in tomato to bipartite begomoviruses Tomato yellow spot virus and Tomato severe rugose virus

    Scientific Electronic Library Online (English)

    Francisco D, Hurtado; Marta A, Gil; Yamila M, Zubiaur; Jorge G, Aguilera; César Augusto D, Xavier; Francisco Murilo, Zerbini Junior; Derly Jose H da, Silva.

    2012-12-01

    Full Text Available Os begomovírus causam doenças de grande importância econômica em diversas culturas, principalmente em regiões tropicais e subtropicais. Juntamente com outras famílias de vírus, os begomovírus têm causado grande prejuízo para os produtores de tomate in natura e para processamento industrial. O objeti [...] vo deste trabalho foi avaliar o comportamento de 11 genótipos resistentes ao Tomato yellow leaf curl virus (TYLCV) frente à infecção pelos begomovírus Tomato yellow spot virus (ToYSV) e Tomato severe rugose virus (ToSRV) em condições de casa-vegetação. A inoculação das plantas foi realizada via biobalística no estádio de duas folhas verdadeiras. A infecção viral confirmou-se pelo desenvolvimento dos sintomas e pela técnica de hibridização dot blot. Selecionaram-se como promissores os genótipos STY2, STY5, STY6 e L7, por não apresentarem sintomas e por terem concentrações virais muito baixas para os dois vírus. O espectro de resistência dos genes Ty-1 e Ty-2 não resultaram efetivos ante as espécies virais empregadas no estudo. As linhagens TY52, H24 e CLN2116B, portadoras destes genes, foram suscetíveis aos vírus ToYSV e ToSRV. Abstract in english Begomoviruses cause diseases of major economic importance in many crops, especially in tropical and subtropical regions. Together with other families of viruses, the begomoviruses cause great damage for producers of fresh and processed tomatoes. The objective of this study was to evaluate the behavi [...] or of 11 resistant genotypes to Tomato yellow leaf curl virus (TYLCV) when infected by the begomoviruses Tomato severe rugose virus (ToSRV) and Tomato yellow spot virus (ToYSV) under greenhouse conditions. Tomato plants were inoculated by biobalistic method when displaying two true leaves. Viral infection was confirmed by visual observation of symptoms and by dot blot hybridization. Genotypes STY2, STY5, STY6 and L7 were selected as promising due to the absence of symptoms and very low viral concentration for both viruses. The resistance genes Ty-1 and Ty-2 were not effective against the viral species used in the study. The lines TY52, H24 and CLN2116B, carrying these genes, exhibited a susceptible behavior.

  4. Detección del iris yellow spot virus en el cultivo de cebolla en Zacatecas, México / Iris yellow spot virus detection in the onion cultivation of Zacatecas, Mexico

    Scientific Electronic Library Online (English)

    Rodolfo, Velásquez-Valle; Manuel, Reveles-Hernández.

    2011-12-01

    Full Text Available El virus de la mancha amarilla del iris (IYSV) es la enfermedad viral de mayor importancia para los cultivos de ajo y cebolla en Zacatecas, México. A finales de mayo de 2010 se encontraron lesiones amarillentas en forma de diamante en hojas y tallo floral de plantas de cebolla en parcelas comerciale [...] s, localizados en los municipios de Loreto, Villa de Cos, Sain Alto, Villanueva, Calera de V. R. y Enrique Estrada pertenecientes al estado de Zacatecas. La prueba DAS-ELISA mostró resultados positivos para este virus en las muestras procedentes de diferentes municipios. Thrips tabaci el único vector conocido del IYSV se encontró en las parcelas comerciales muestreadas, aunque la presencia de Frankliniella occidentalis también fue reconocida en una parcela comercial de cebolla, cuyas muestras resultaron positivas a IYSV. No se encontró diferencia entre la altura, número de hojas, peso y diámetro de bulbo de plantas sin lesiones y aquellas con diferente número de lesiones de IYSV. Abstract in english The yellow spot virus Iris (IYSV) is the most important viral disease for garlic and onion crops in Zacatecas, Mexico. In late May 2010, yellow lesions were found in diamond-shaped leaves and flower stem of onion plants in commercial fields located in the municipalities of Loreto, Villa de Cos, Sain [...] Alto, Villanueva, Calera de V. R. and Enrique Estrada from the State of Zacatecas. The DAS-ELISA test was positive for the virus in samples from different municipalities. Thrips tabaci the only known vector of IYSV was found in sampled commercial parcels, although, the presence of Frankliniella occidentalis was also recognized in a commercial plot of onions, whose samples were positive for IYSV too. No difference was found between height, leaf number, bulb weight or diameter of the plants without lesions and those with different numbers of IYSV lesions.

  5. 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 ?-dioxygenase-1 (?-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 ?-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. PMID:23487466

  6. Therapeutic effect of two fluoride varnishes on white spot lesions: a randomized clinical trial

    Directory of Open Access Journals (Sweden)

    Jainara Maria Soares Ferreira

    2009-12-01

    Full Text Available The aim of this randomized clinical trial study was to evaluate the therapeutic effect of two varnish formulations (G1 = 5% NaF, G2 = 6% NaF + 6% CaF2 on the remineralization of white spot lesions (WSL. The sample was composed of 15 (7- to 12-year-old children with 45 active WSL in anterior permanent teeth. The children were randomly divided into two groups providing 22 lesions for G1 and 23 for G2. The children were submitted to weekly varnish applications 4 times. The WSL were evaluated twice: baseline and on week 4. Maximum lesion dimensions (mesiodistal and incisogingival were measured in millimeters and classified in four grades of size. WSL were also assessed regarding lesion activity by one calibrated examiner. The Pearson chi-square and Fisher's exact tests were used (P 0.01. It was concluded that after 4 applications the two varnish formulations tested produced similar clinical effects, indicating the reduction and the control of carious activity in most WSL.

  7. On the formation of white-spot defects in a superalloy VAR ingot

    Energy Technology Data Exchange (ETDEWEB)

    Grignard, Jean Francois; Soller, Aude; Jourdan, Julien; Bellot, Jean-Pierre; Jardy, Alain [Institut Jean Lamour (UMR 7198), Ecole des Mines de Nancy, Parc de Saurupt, Nancy Cedex (France)

    2011-07-15

    For the production of nickel-based superalloys for the aerospace industry, strict control of the macrostructure of the product is necessary to avoid the appearance of potentially fatal defects. Our study focuses on the prevention of ''white spots'' in the alloy IN 718. These defects, which are small volumes of a few millimeters of characteristic length, are depleted in niobium. They are known to result from the fall of metal fragments in the liquid pool during VAR processing. According to their history in the liquid metal, these fragments could not being remelted before being trapped in the mushy zone and then give rise to defects. A model calculates the heat transfer in such a precursor to simulate its melting during his stay in the bath. The validation of the predicted melting kinetics requires a series of immersive experiences of synthetic defects in a metal bath. The model and experiments have demonstrated the initial solidification of a layer of metal around the precursor. (Copyright copyright 2011 WILEY-VCH Verlag GmbH and Co. KGaA, Weinheim)

  8. Factors Affecting the Formation, Severity and Location of White Spot Lesions during Orthodontic Treatment with Fixed Appliances

    OpenAIRE

    Khaled Khalaf

    2014-01-01

    Objectives: The purpose of the present study was to investigate factors affecting the formation, severity and location of white spot lesions in patients completing fixed appliance therapy. Material and Methods: A total of 45 patients (19 males and 26 females, mean age 15.81 years, standard deviation 2.89 years) attending consecutively Aberdeen Dental Hospital (ADH) between January and June 2013 to have their fixed appliances removed were given a questionnaire to elicit information regardi...

  9. Null mutation of endothelin receptor type B gene in spotting lethal rats causes aganglionic megacolon and white coat color.

    OpenAIRE

    Gariepy, C. E.; Cass, D. T.; Yanagisawa, M.

    1996-01-01

    Mutations in the gene encoding the endothelin receptor type B (EDNRB) produce congenital aganglionic megacolon and pigment abnormalities in mice and humans. Here we report a naturally occurring null mutation of the EDNRB gene in spotting lethal (sl) rats, which exhibit aganglionic megacolon associated with white coat color. We found a 301-bp deletion spanning the exon 1-intron 1 junction of the EDNRB gene in sl rats. A restriction fragment length polymorphism caused by this deletion perfectly...

  10. Porites white patch syndrome: associated viruses and disease physiology

    Science.gov (United States)

    Lawrence, S. A.; Davy, J. E.; Wilson, W. H.; Hoegh-Guldberg, O.; Davy, S. K.

    2015-03-01

    In recent decades, coral reefs worldwide have undergone significant changes in response to various environmental and anthropogenic impacts. Among the numerous causes of reef degradation, coral disease is one factor that is to a large extent still poorly understood. Here, we characterize the physiology of white patch syndrome (WPS), a disease affecting poritid corals on the Great Barrier Reef. WPS manifests as small, generally discrete patches of tissue discolouration. Physiological analysis revealed that chlorophyll a content was significantly lower in lesions than in healthy tissues, while host protein content remained constant, suggesting that host tissue is not affected by WPS. This was confirmed by transmission electron microscope (TEM) examination, which showed intact host tissue within lesions. TEM also revealed that Symbiodinium cells are lost from the host gastrodermis with no apparent harm caused to the surrounding host tissue. Also present in the electron micrographs were numerous virus-like particles (VLPs), in both coral and Symbiodinium cells. Small (<50 nm diameter) icosahedral VLPs were significantly more abundant in coral tissue taken from diseased colonies, and there was an apparent, but not statistically significant, increase in abundance of filamentous VLPs in Symbiodinium cells from diseased colonies. There was no apparent increase in prokaryotic or eukaryotic microbial abundance in diseased colonies. Taken together, these results suggest that viruses infecting the coral and/or its resident Symbiodinium cells may be the causative agents of WPS.

  11. Proteomic Analysis of Frankliniella occidentalis and Differentially Expressed Proteins in Response to Tomato Spotted Wilt Virus Infection

    OpenAIRE

    Badillo-Vargas, I. E.; Rotenberg, D.; Schneweis, D. J.; Hiromasa, Y.; Tomich, J M; Whitfield, A E

    2012-01-01

    Tomato spotted wilt virus (TSWV) is transmitted by Frankliniella occidentalis in a persistent propagative manner. Despite the extensive replication of TSWV in midgut and salivary glands, there is little to no pathogenic effect on F. occidentalis. We hypothesize that the first-instar larva (L1) of F. occidentalis mounts a response to TSWV that protects it from pathogenic effects caused by virus infection and replication in various insect tissues. A partial thrips transcriptome was generated us...

  12. Resistência de cultivares e linhagens de tomateiro a Tomato chlorotic spot virus e a Potato virus Y / Resistance of tomato lines and cultivars to Tomato chlorotic spot virus and Potato virus Y

    Scientific Electronic Library Online (English)

    André L., Lourenção; Walter J., Siqueira; Arlete M. T., Melo; Silvia R.L., Palazzo; Paulo C.T., Melo; Addolorata, Colariccio.

    2005-12-01

    Full Text Available Linhagens avançadas do programa de melhoramento do tomateiro (Lycopersicon esculentum) do IAC foram avaliadas em condições de campo em Campinas (SP) para resistência a tospovírus e a potyvírus, nos anos agrícolas 2002/2003 e 2003/2004, respectivamente. No primeiro ano, a única espécie de tospovírus [...] que ocorreu na área experimental foi Tomato chlorotic spot virus (TCSV). As sete linhagens do grupo IAC exibiram baixa porcentagem de plantas sintomáticas em duas avaliações, com médias abaixo de 28%; as cultivares testadas mostraram-se altamente suscetíveis, com médias acima de 85%, à exceção de 'Franco', que apresentou cerca de 55% de infecção. No segundo experimento, conduzido em 2003/2004, dez linhagens do grupo IAC foram comparadas com cinco cultivares de polinização aberta e híbridos F1, além do acesso LA-444-1 de L. peruvianum. Nesse experimento, por meio de testes biológicos e sorológicos, verificou-se ocorrência generalizada de Potato virus Y (PVY). Foi determinado o percentual de plantas com sintomas e avaliada a intensidade dos sintomas mediante uso de escala de notas. Com base nos dois critérios, verificou-se que LA-444-1 apresenta alta resistência a PVY, que 'Tyrade' exibe comportamento intermediário, enquanto todos os demais genótipos demonstram alta suscetibilidade ao vírus. O comportamento dos genótipos avaliados neste trabalho mostra a necessidade de se considerar, nos programas de melhoramento do tomateiro, a introgressão de fatores de resistência não só a vírus de importância atual nas regiões produtoras, como geminivírus, mas também a outros vírus potencialmente nocivos à cultura, como tospovírus e potyvírus. Abstract in english Advanced breeding lines from the IAC tomato breeding program and several tomato (Lycopersicon esculentum) cultivars and F1 hybrids were screened for tospovirus and potyvirus resistance under field conditions, at Campinas, São Paulo State, Brazil, during the 2002/2003 and 2003/2004 growing seasons. D [...] uring the first season, only Tomato chlorotic spot virus (TCSV) was detected in plants of the experimental area. On both evaluations, all seven lines (IAC group) were resistant to TCSV, with under 28% of infected plants. The tomato cultivars and hybrids were highly susceptible, with greater than 85% of infected plants, except for 'Franco', with 55% of infected plants. During the 2003/2004 growing season, the number of IAC lines evaluated was raised from seven to ten, and they were compared to five cultivars/F1 hybrids and to L. peruvianum LA-444-1. On this experiment, only Potato virus Y (PVY) was detected in plants. Evaluations were carried out using a symptom intensity scale and ELISA. Considering both criteria, it was verified that only LA-444-1 displayed high resistance to PVY. In addition, 'Tyrade' displayed an intermediate behavior while all other lines, cultivars and hybrids behaved as susceptible to this potyvirus. These results highlight the need of introgressing resistance to multiple viruses in tomato breeding programs, taking into consideration the economical importance and relative incidence of each virus in different geographical regions and natural variations on incidence from year to year.

  13. Resistência de cultivares e linhagens de tomateiro a Tomato chlorotic spot virus e a Potato virus Y Resistance of tomato lines and cultivars to Tomato chlorotic spot virus and Potato virus Y

    Directory of Open Access Journals (Sweden)

    André L. Lourenção

    2005-12-01

    Full Text Available Linhagens avançadas do programa de melhoramento do tomateiro (Lycopersicon esculentum do IAC foram avaliadas em condições de campo em Campinas (SP para resistência a tospovírus e a potyvírus, nos anos agrícolas 2002/2003 e 2003/2004, respectivamente. No primeiro ano, a única espécie de tospovírus que ocorreu na área experimental foi Tomato chlorotic spot virus (TCSV. As sete linhagens do grupo IAC exibiram baixa porcentagem de plantas sintomáticas em duas avaliações, com médias abaixo de 28%; as cultivares testadas mostraram-se altamente suscetíveis, com médias acima de 85%, à exceção de 'Franco', que apresentou cerca de 55% de infecção. No segundo experimento, conduzido em 2003/2004, dez linhagens do grupo IAC foram comparadas com cinco cultivares de polinização aberta e híbridos F1, além do acesso LA-444-1 de L. peruvianum. Nesse experimento, por meio de testes biológicos e sorológicos, verificou-se ocorrência generalizada de Potato virus Y (PVY. Foi determinado o percentual de plantas com sintomas e avaliada a intensidade dos sintomas mediante uso de escala de notas. Com base nos dois critérios, verificou-se que LA-444-1 apresenta alta resistência a PVY, que 'Tyrade' exibe comportamento intermediário, enquanto todos os demais genótipos demonstram alta suscetibilidade ao vírus. O comportamento dos genótipos avaliados neste trabalho mostra a necessidade de se considerar, nos programas de melhoramento do tomateiro, a introgressão de fatores de resistência não só a vírus de importância atual nas regiões produtoras, como geminivírus, mas também a outros vírus potencialmente nocivos à cultura, como tospovírus e potyvírus.Advanced breeding lines from the IAC tomato breeding program and several tomato (Lycopersicon esculentum cultivars and F1 hybrids were screened for tospovirus and potyvirus resistance under field conditions, at Campinas, São Paulo State, Brazil, during the 2002/2003 and 2003/2004 growing seasons. During the first season, only Tomato chlorotic spot virus (TCSV was detected in plants of the experimental area. On both evaluations, all seven lines (IAC group were resistant to TCSV, with under 28% of infected plants. The tomato cultivars and hybrids were highly susceptible, with greater than 85% of infected plants, except for 'Franco', with 55% of infected plants. During the 2003/2004 growing season, the number of IAC lines evaluated was raised from seven to ten, and they were compared to five cultivars/F1 hybrids and to L. peruvianum LA-444-1. On this experiment, only Potato virus Y (PVY was detected in plants. Evaluations were carried out using a symptom intensity scale and ELISA. Considering both criteria, it was verified that only LA-444-1 displayed high resistance to PVY. In addition, 'Tyrade' displayed an intermediate behavior while all other lines, cultivars and hybrids behaved as susceptible to this potyvirus. These results highlight the need of introgressing resistance to multiple viruses in tomato breeding programs, taking into consideration the economical importance and relative incidence of each virus in different geographical regions and natural variations on incidence from year to year.

  14. Immunological-based assays for specific detection of shrimp viruses

    OpenAIRE

    Chaivisuthangkura, Parin; Longyant, Siwaporn; Sithigorngul, Paisarn

    2014-01-01

    Among shrimp viral pathogens, white spot syndrome virus (WSSV) and yellow head virus (YHV) are the most lethal agents, causing serious problems for both the whiteleg shrimp, Penaeus (Litopenaeus) vannamei, and the black tiger shrimp, Penaeus (Penaeus) monodon. Another important virus that infects P. vannamei is infectious myonecrosis virus (IMNV), which induces the white discoloration of affected muscle. In the cases of taura syndrome virus and Penaeus stylirostris densovirus (PstDNV; formerl...

  15. Children with severe early childhood caries: streptococci genetic strains within carious and white spot lesions

    Directory of Open Access Journals (Sweden)

    Kenneth Gilbert

    2014-10-01

    Full Text Available Background and objectives: Mutans streptococci (MS are one of the major microbiological determinants of dental caries. The objectives of this study are to identify distinct MS and non-MS streptococci strains that are located at carious sites and non-carious enamel surfaces in children with severe early childhood caries (S-ECC, and assess if cariogenic MS and non-cariogenic streptococci might independently exist as primary bacterial strains on distinct sites within the dentition of individual children. Design: Dental plaque from children (N=20; aged 3–6 with S-ECC was collected from carious lesions (CLs, white spot lesions (WSLs and non-carious enamel surfaces. Streptococcal isolates (N=10–20 from each site were subjected to polymerase chain reaction (PCR to identify MS, and arbitrarily primed-PCR for assignment of genetic strains. Primary strains were identified as ?50% of the total isolates surveyed at any site. In several cases, strains were characterized for acidurity using ATP-driven bioluminescence and subjected to PCR-determination of potential MS virulence products. Identification of non-MS was determined by 16S rRNA gene sequencing. Results: Sixty-four independent MS or non-MS streptococcal strains were identified. All children contained 1–6 strains. In many patients (N=11, single primary MS strains were identified throughout the dentition. In other patients (N=4, primary MS strains were identified within CLs that were distinct from primary strains found on enamel. Streptococcus gordonii strains were identified as primary strains on enamel or WSLs in four children, and in general were less aciduric than MS strains. Conclusions: Many children with S-ECC contained only a single primary MS strain that was present in both carious and non-carious sites. In some cases, MS and non-cariogenic S. gordonii strains were found to independently exist as dominant strains at different locations within the dentition of individual children, and the aciduric potential of these strains may influence susceptibility in the development of CLs.

  16. Virus infection decreases the attractiveness of white clover plants for a non-vectoring herbivore

    DEFF Research Database (Denmark)

    van Mölken, Tamara; Caluwe, Hannie de

    2012-01-01

    Plant pathogens and insect herbivores are prone to share hosts under natural conditions. Consequently, pathogen-induced changes in the host plant can affect herbivory, and vice versa. Even though plant viruses are ubiquitous in the field, little is known about plant-mediated interactions between viruses and non-vectoring herbivores. We investigated the effects of virus infection on subsequent infestation by a non-vectoring herbivore in a natural genotype of Trifolium repens (white clover). We tested whether infection with White clover mosaic virus (WClMV) alters (1) the effects of fungus gnat feeding on plant growth, (2) the attractiveness of white clover for adult fungus gnat females, and (3) the volatile emission of white clover plants. We observed only marginal effects of WClMV infection on the interaction between fungus gnat larvae and white clover. However, adult fungus gnat females clearly preferred non-infected over WClMV-infected plants. Non-infected and virus-infected plants could easily be discriminated based on their volatile blends, suggesting that the preference of fungus gnats for non-infected plants may be mediated by virus-induced changes in volatile emissions. The compound beta-caryophyllene was exclusively detected in the headspace of virus-infected plants and may hence be particularly important for the preference of fungus gnat females. Our results demonstrate that WClMV infection can decrease the attractiveness of white clover plants for fungus gnat females. This suggests that virus infections may contribute to protecting their hosts by decreasing herbivore infestation rates. Consequently, it is conceivable that viruses play a more beneficial role in plant-herbivore interactions than generally thought.

  17. Factors Affecting the Formation, Severity and Location of White Spot Lesions during Orthodontic Treatment with Fixed Appliances

    Directory of Open Access Journals (Sweden)

    Khaled Khalaf

    2014-04-01

    Full Text Available Objectives: The purpose of the present study was to investigate factors affecting the formation, severity and location of white spot lesions in patients completing fixed appliance therapy. Material and Methods: A total of 45 patients (19 males and 26 females, mean age 15.81 years, standard deviation 2.89 years attending consecutively Aberdeen Dental Hospital (ADH between January and June 2013 to have their fixed appliances removed were given a questionnaire to elicit information regarding their dental care and diet. They were then examined clinically as well as their pre-treatment photographs to record treatment data and white spot lesion (WSL location and severity using a modified version of Universal Visual Scale for Smooth Surfaces (UniViSS Smooth. Absolute risk (AR and risk ratios (RR were also calculated. Results: The incidence of at least one WSL observed in patients was 42%, with males displaying a higher incidence than females. The highest incidence of WSLs was recorded on the maxillary canines and lateral incisors, and on the maxillary and mandibular premolars and first molars. The gingival areas of the maxillary and mandibular teeth were the most affected surfaces. Significant (P < 0.05 relationships were found between the presence of WSLs and the following factors: poor oral hygiene (OH, males, increased treatment length, lack of use of fluoride supplements, use of carbonated soft drinks and/or fruit juices and the use of sugary foods. Poor OH posed the highest risk of developing WSL (RR = 8.55. Conclusions: 42% of patients have developed white spot lesions during fixed appliance therapy. Various contributing risk factors were identified with the greatest risk posed by a poor oral hygiene.

  18. Proteomic analysis of Frankliniella occidentalis and differentially expressed proteins in response to tomato spotted wilt virus infection.

    Science.gov (United States)

    Badillo-Vargas, I E; Rotenberg, D; Schneweis, D J; Hiromasa, Y; Tomich, J M; Whitfield, A E

    2012-08-01

    Tomato spotted wilt virus (TSWV) is transmitted by Frankliniella occidentalis in a persistent propagative manner. Despite the extensive replication of TSWV in midgut and salivary glands, there is little to no pathogenic effect on F. occidentalis. We hypothesize that the first-instar larva (L1) of F. occidentalis mounts a response to TSWV that protects it from pathogenic effects caused by virus infection and replication in various insect tissues. A partial thrips transcriptome was generated using 454-Titanium sequencing of cDNA generated from F. occidentalis exposed to TSWV. Using these sequences, the L1 thrips proteome that resolved on a two-dimensional gel was characterized. Forty-seven percent of the resolved protein spots were identified using the thrips transcriptome. Real-time quantitative reverse transcriptase PCR (RT-PCR) analysis of virus titer in L1 thrips revealed a significant increase in the normalized abundance of TSWV nucleocapsid RNA from 2 to 21 h after a 3-h acquisition access period on virus-infected plant tissue, indicative of infection and accumulation of virus. We compared the proteomes of infected and noninfected L1s to identify proteins that display differential abundances in response to virus. Using four biological replicates, 26 spots containing 37 proteins were significantly altered in response to TSWV. Gene ontology assignments for 32 of these proteins revealed biological roles associated with the infection cycle of other plant- and animal-infecting viruses and antiviral defense responses. Our findings support the hypothesis that L1 thrips display a complex reaction to TSWV infection and provide new insights toward unraveling the molecular basis of this interaction. PMID:22696645

  19. Differences in organotin accumulation in relation to life history in the white-spotted charr Salvelinus leucomaenis

    International Nuclear Information System (INIS)

    Research highlights: ? Otolith Sr:Ca ratios in sea-run type were higher than those in freshwater-residents. ? TBT and TPT concentrations in sea-run type were higher than those in freshwater-residents. ? Sea-run type have higher risk of TBT and TPT than freshwater-residents in white-spotted charr. - Abstract: To examine the accumulation pattern of organotins (OTs) in relation to the migration of diadromous fish, tributyltin (TBT) and triphenyltin (TPT) and their derivatives were determined in the muscle tissue of both sea-run (anadromous) and freshwater-resident (nonanadromous) types of the white-spotted charr Salvelinus leucomaenis. Ontogenic changes in otolith strontium (Sr) and calcium (Ca) concentrations were examined along life history transect to discriminate migration type. Mean Sr:Ca ratio from the core to the edge of the otolith in sea-run individuals was significantly higher than those in freshwater-resident one. There were no significant correlations in S. leucomaenis between OT accumulation and various biological characteristics. It is noteworthy that TBT and TPT concentrations in sea-run type were significantly higher than those in freshwater-resident individuals, although they are both of the same species. These results suggest that sea-run S. leucomaenis have a higher ecological risk of OT exposure than freshwater-residents during their life histories.

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

    OpenAIRE

    Ivana ?eki?; Aleksandra Bulaji?; Ana Vu?urovi?; Danijela Risti?; Branka Krsti?

    2008-01-01

    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 a critical moment upon which the reliability o...

  1. HEPATITIS B VIRUS DNA CAN BE AMPLIFIED DIRECTLY FROM DRIED BLOOD SPOT ON FILTER PAPER

    Directory of Open Access Journals (Sweden)

    Naglaa Fathy Alhusseini

    2012-01-01

    Full Text Available Hepatitis B Virus (HBV infections represent a major public health problem because of the ability of HBV to cause a chronic carrier state. Even though chronic carriers remain largely asymptomatic, a large number of these individuals subsequently develop cirrhosis and primary hepatocellular carcinomas. Dried Blood Spot (DBS samples are a simple and inexpensive sampling method, especially useful for blood collection in resource poor settings with limited access to diagnostic facilities. The main advantage of DBS samples over routine blood samples is that only a small quantity of blood, is required. They are easy to obtain, stable and can be transported to a reference laboratory at minimal cost. This study was to evaluate the feasibility of DBS samples for direct amplification of HBV DNA bypassing nucleic acid extraction. Results obtained from DBS samples were compared from those from plasma by routine molecular technique and also with those from whole blood. On the whole results for DBS, whole blood and plasma samples for HBV-DNA semi quantitative PCR monitoring, demonstrated very good agreement. This study also represents the first report in Egypt to evaluate the use of DBS to direct amplification of HBV-DNA and concluded that the use of DBS for direct amplification of HBV DNA without nucleic acid extraction was reliable, specific, sensitive, cheap and appropriate method to monitor the HBV infected patients."

  2. 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. PMID:15551034

  3. Mineral content of ionomer cements and preventive effect of these cements against white spot lesions around restorations

    Scientific Electronic Library Online (English)

    Lilian Fernanda Santos, PAIVA; Tatiana Kelly da Silva, FIDALGO; Lucianne Cople, MAIA.

    2014-08-29

    Full Text Available This study evaluated the ion exchange at the material/enamel interface and the preventive effect of restorative materials submitted to cariogenic challenge against white spot. Restorations in enamel/dentin of bovine teeth were performed with composite resin (Filtek™ Z250 – control group) and glass-i [...] onomers cements - GICs (Ionomaster R™ and Fuji IX™ - experimental groups). Samples were grouped and submitted to neutral saliva (n = 15) or pH-cycling regimen (n = 15). After eight days of pH cycling, material/enamel interfaces were analyzed by EDX in order to determine the differences (p 0.05) versus the experimental groups. Ca and P content were higher in enamel than in restorative materials. After pH cycling, the GIC enamel bulk showed a significantly higher Sr content compared with the composite resin (p

  4. Immune responses of orange-spotted grouper, Epinephelus coioides, against virus-like particles of betanodavirus produced in Escherichia coli.

    Science.gov (United States)

    Lai, Yu-Xiong; Jin, Bao-Lei; Xu, Yu; Huang, Li-Jie; Huang, Run-Qing; Zhang, Yong; Kwang, Jimmy; He, Jian-Guo; Xie, Jun-Feng

    2014-01-15

    Betanodaviruses are the causative agents of viral nervous necrosis (VNN), a serious disease of cultured marine fish worldwide. Virus-like particles (VLPs) are one of the good novel vaccine candidates to control this disease. Until now, betanodavirus vaccine studies mainly focused on the humoral immune response and mortality after virus challenge. However, little is known about the activation of genes responsible for cellular and innate immunity by vaccines. In the present study, VLPs of orange-spotted grouper nervous necrosis virus (OGNNV) were produced in prokaryotes and their ability to enter Asian sea bass cells was the same as native virus, suggesting that they possess a similar structure to OGNNV. VLPs immunogenicity was then determined by intramuscularly vaccinating Epinephelus coioides at different concentrations (1.5 or 15 ?g g(-1) fish body weight, FBW) and immunizing frequencies (administration once, twice and thrice). A single vaccination with the dosage of 1.5 ?g g(-1) FBW is enough to provoke high titer antibodies (average 3 fold higher than that of negative control) with strong neutralizing antibody titer as early as 1 week post immunization. Furthermore, quantitative PCR analysis revealed that eleven genes associated with humoral, cellular and innate immunities were up-regulated in the liver, spleen and head kidney at 12h post immunization, correlating with the early antibody response. In conclusion, we demonstrated that VLP vaccination induced humoral immune responses and activated genes associated with cellular and innate immunity against betanodavirus infection in orange-spotted grouper. PMID:24252246

  5. Tomato spotted wilt virus: agente causal de la marchitez del Miguelito (Zinnia elegans Jacquin) en el Estado de Morelos, México / Tomato spotted wilt virus: causal agent of wilt in "Miguelito" (Zinnia elegans Jacquin) in Morelos, Mexico

    Scientific Electronic Library Online (English)

    Ma. Valeria, Morales-Díaz; Salomé, Alcacio-Rangel; Rodolfo, De La Torre-Almaraz.

    2008-05-01

    Full Text Available Se observaron síntomas de moteado, manchas necróticas anulares, enanismo y marchitez severa en plantas de miguelito (Zinnia elegans Jacquin) (Fam: Compositae) cultivadas en viveros comerciales del Estado de Morelos, México. Por tanto, el objetivo del presente trabajo fue determinar la etiología de e [...] sta enfermedad. Se identificó al virus marchitez manchada del tomate (Tomato spotted wilt virus. TSWV) como el agente causal de la enfermedad del miguelito con base en los ensayos de transmisión mecánica en plantas indicadoras y de patogenicidad en plántulas sanas de Z. elegans, cultivadas desde semilla en el invernadero. En las plantas el virus causó los síntomas de moteado, manchas necróticas en forma de anillos concéntricos, enanismo y marchitez, idénticos a los observados en plantas de miguelito cultivados en condiciones de campo. Se detectó por serología (DAS-ELISA) sólo al TSWV en plantas de miguelito con síntomas de marchitez procedentes de viveros comerciales del Estado de Morelos, en las plantas indicadoras usadas para separar al virus y en las plantas de miguelito producidas desde semilla inoculadas con el aislamiento puro de TSWV en el invernadero. La identidad taxonómica del TSWV se confirmó mediante secuenciación directa de los productos de la RT-PCR. Las secuencias obtenidas del TSWV de muestras de campo de miguelito (número de acceso EF067862) y del mismo virus separado de N. rustica (número de acceso EF067863) mostraron una homología en el fragmento amplificado del gen de la proteína de la cápside del TSWV del 100% entre ellas y del 93% con las existentes en la base de datos de lNCBI/GenBank de aislamientos del TSWV ampliamente distribuidos en el mundo. Este trabajo es el primer reporte de la presencia del TSWV en Z. elegans en México y se describen dos métodos moleculares para diagnosticar e identificar este virus. Abstract in english Mottle symptoms, necrotic ring spots, dwarfism, and severe wilt were observed on "miguelito" (Zinnia elegans Jacquin) (Fam: Comositae) plants cultivated in commercial nurseries in the State of Morelos, México. Therefore, this study was conducted to determine the etiology of the disease. Tomato spott [...] ed wilt virus (TSWV) was identified as the causal agent of the disease in miguelito by mechanical transmission tests on indicator plants and pathogenicity tests on healthy Z. elegans seedlings cultivated from seed in a greenhouse. On these plants the virus caused mottled symptoms, necrotic spots in the form of concentric rings, dwarfism and wilt, identical to those observed in miguelito plants cultivated in field conditions. Through serology (DAS-ELISA) only TSWV was detected in miguelito plants with wilt symptoms from commercial nurseries in the State of Morelos, in indicator plants used to separate the virus, and in miguelito plants produced from seed and inoculated with a pure isolate of TSWV in the greenhouse. Taxonomical identity of TSWV was confirmed by direct sequencing of the RT-PCR products. The TSWV sequences obtained from field samples of miguelito (access number EF067862) and the same virus separated from N. rustica (access number EF067863) showed 100% homology in the amplified fragment of the capsid protein gene of TSWV, while with the samples of TSWV isolates widely distributed over the world existing in the INCBI/GeneBank there was 93% homology. This paper is the first report of the presence of TSWV in Z. elegans in México, and two molecular methods for diagnosing and identifying this virus are described.

  6. Control of White Spot Lesion Adjacent to Orthodontic Bracket with Use of Fluoride Varnish or Chlorhexidine Gel

    Science.gov (United States)

    Restrepo, Manuel; Bussaneli, Diego G.; Jeremias, Fabiano; Cordeiro, Rita C. L.; Magalhães, Ana C.; Palomari Spolidorio, Denise M.; Santos-Pinto, Lourdes

    2015-01-01

    The aims of this study were to compare the effectiveness of fluoride varnish and chlorhexidine gel in controlling white spot lesions (WSLs) adjacent to orthodontic brackets and to compare the ability of Quantitative Light-Induced Fluorescence (QLF) to measure mineral uptake with that of transverse microradiography (TMR). Thirty premolars with artificially induced WSLs were randomly assigned to three groups: (1) two applications of 5% NaF-varnish (F), with one-week interval, (2) two applications of 2% chlorhexidine gel (CHX), with one-week interval, and (3) control (CO), no treatment. QLF was used to measure changes in fluorescence before and after caries induction, 1 week after each application and 1, 2, and 3 months after the last application of F or CHX. TMR was performed to quantify lesion depth and mineral content after caries induction to evaluate the effects of F, CHX, and CO 3 months after the last application of agents. The data were analyzed by repeated measures ANOVA and Tukey's test. All treatments increased the mineral content during the experimental period; however, F induced faster remineralization than CHX. The correlation between QLF and TMR was significantly moderate. Two applications of fluoride varnish or 2% chlorhexidine gel at one-week intervals were effective in controlling WSLs. PMID:25973442

  7. White matter tract injury and cognitive impairment in human immunodeficiency virus–infected individuals

    OpenAIRE

    Gongvatana, Assawin; Schweinsburg, Brian C.; Taylor, Michael J.; Theilmann, Rebecca J.; Letendre, Scott L.; Alhassoon, Omar M.; Jacobus, Joanna; Woods, Steven P.; Jernigan, Terry L.; Ellis, Ronald J.; Frank, Lawrence R.; Grant, Igor

    2009-01-01

    Approximately half of those infected with the human immunodeficiency virus (HIV) exhibit cognitive impairment, which has been related to cerebral white matter damage. Despite the effectiveness of antiretroviral treatment, cognitive impairment remains common even in individuals with undetectable viral loads. One explanation for this may be subtherapeutic concentrations of some antiretrovirals in the central nervous system (CNS). We utilized diffusion tensor imaging and a comprehensive neuropsy...

  8. Comparison of organotin accumulation on the white-spotted charr Salvelinus leucomaenis between sea-run and freshwater-resident types

    Science.gov (United States)

    Ohji, Madoka; Harino, Hiroya; Arai, Takaomi

    2011-01-01

    To examine the accumulation pattern of organotin compounds (OTs) in relation to the migration of diadromous fish, tributyltin (TBT) and triphenyltin (TPT) compounds and their derivatives were determined in the muscle tissue of both sea-run (anadromous) and freshwater-resident (nonanadromous) types of the white-spotted charr Salvelinus leucomaenis. There were generally no significant correlations between the TBT and TPT accumulation and various biological characteristics such as the total length (TL), body weight (BW), age and sex in S. leucomaenis. It is noteworthy that the TBT and TPT concentrations in sea-run white-spotted charr were significantly higher than in freshwater-resident individuals, although they are intraspecies. These results suggest that the sea-run S. leucomaenis has a higher ecological risk of TBT and TPT exposure than the freshwater-residents during their life history.

  9. Postsynaptic Alpha 2-Adrenoceptors Mediate Melanosome Aggregation in Melanophores of the White-Spotted Rabbitfish (Siganus canaliculatus)

    OpenAIRE

    M.H. Amiri

    2009-01-01

    The present investigation was undertaken to study the nature of neuro-melanophore junction in the white-spotted rabbit fish Siganus cancliculatus. In vitro experiments using split fin preparation indicated that melanophores of S. cancliculatus are highly responsive to potassium ions and adrenergic agonists. Potassium ions and the adrenergic agonists induced prompt melanosome aggregation that could be competitively blocked by yohimbine (alpha-2 specific adrenergic antagonist) and phento...

  10. Persistence of hepatitis C virus in a white population: associations with human leukocyte antigen class 1.

    LENUS (Irish Health Repository)

    Fanning, Liam J

    2012-02-03

    The aim of this study was to define novel associations between human leukocyte antigen (HLA) class 1 alleles and persistence or clearance of the hepatitis C virus (HCV) in a white population. All individuals in the study were seropositive for anti-HCV antibodies. Viral status was determined by the Roche HCV Amplicor test. HLA-A, -B, -C allelic group profile was molecularly defined by reverse line probe hybridization. The strongest individual allelic group associations with persistent HCV infection were HLA A*11 (p = 0.044) and Cw*04 (p = 0.006). However, only the HLA C*04 association survived correction for multiple comparisons. Further analysis of alleles in linkage with HLA Cw*04 revealed that the haplotype HLA A*11, Cw*04 was present in 11 individuals, 10 of whom were viremic (p = 0.05). No gene dosage effect was observed. No association between HLA class 1 allelic groups and aviremia and virus load was evident in this white population. HLA B*44 is associated with low virus load in human immunodeficiency virus disease, but this association was not evident in this HCV-infected population. Novel HLA class 1 alleles associated with persistence of HCV have been identified.

  11. First Report of Impatiens necrotic spot virus in Blackberry in the United States

    Science.gov (United States)

    Blackberry yellow vein disease (BYVD) has emerged in the last six years in the south and southeastern United States and can be devastating. It is caused by the synergistic effect of several viruses, with Blackberry yellow vein associated virus being a consistent component of the disorder. In an eff...

  12. Viral MicroRNAs Targeting Virus Genes Promote Virus Infection in Shrimp In Vivo

    OpenAIRE

    He, Yaodong; Kai YANG; Zhang, Xiaobo

    2014-01-01

    Viral microRNAs (miRNAs), most of which are characterized in cell lines, have been found to play important roles in the virus life cycle to avoid attack by the host immune system or to keep virus in the latency state. Viral miRNAs targeting virus genes can inhibit virus infection. In this study, in vivo findings in Marsupenaeus japonicus shrimp revealed that the viral miRNAs could target virus genes and further promote the virus infection. The results showed that white spot syndrome virus (WS...

  13. Effect of infection by chlorotic spot virus on 14CO2 fixation in leaves of groundnut Arachis hypogea L

    International Nuclear Information System (INIS)

    Photosynthetic incorporation of 14CO2 into leaves of groundnut infected by chlorotic spot virus (GCSV) was slightly more at stages 2 and 5 less at stage 4 as compared to control. 14C incorporation into the alcohol soluble fraction of infected leaves followed the same trend as total 14CO2 fixation but in the alcohol-insoluble fraction the same was less at all the sampled stages. 14C in the alcohol-soluble fraction of fed leaves of both types (stage 5) decreased with time along with simultaneous increase in alcohol-insoluble fraction. The proportion of 14C incorporated into organic acids, amino acids and sugars was same in both the samples at stage 2, greater into organic and amino acids and less into sugars at stages 4 and 5, and at 12 and 24 hr time periods of stage 5 of virus infected leaves when compared to healthy ones. 14C incorporated into total sugars and organic acids of infected leaves followed that of total 14C fixation, and varied in individual sugars and organic acids. 14C in sugars of both type of leaves decreased with time and with simultaneous increase in organic and amino acids. 14C incorporated into virus infected leaf proteins was more when compared to healthy leaves. (auth.)

  14. Replication of white spot syndrome virus (WSSV) in the polychaete Dendronereis spp

    OpenAIRE

    Desrina, H.; Verreth, J.A.J.; S.B. Prayitno; Rombout, J.H.W.M.; J. M. Vlak; Verdegem, M. C. J.

    2013-01-01

    This study investigated whether WSSV replicates in naturally infected Dendronereis spp., a common polychaete (Nereididae) species in shrimp ponds in Indonesia. To detect WSSV replication, (i) immunohistochemistry (IHC) using a monoclonal antibody against WSSV VP28 protein and (ii) nested RT-PCR using specific primers set for the vp28 gene to detect WSSV-specific mRNA were applied. WSSV immunoreactive-nuclei were detected in the gut epithelium of the polychaete and WSSV mRNA was detected with ...

  15. White spot syndrome virus VP51 interact with ribosomal protein L7 of Litopenaeus vannamei.

    Science.gov (United States)

    Liu, Qing-Hui; Ma, Fang-Fang; Guan, Guang-Kuo; Wang, Xiu-Fang; Li, Chen; Huang, Jie

    2015-05-01

    The interaction between viral structural proteins and host plays key functions in viral infection. In previous studies, most research have been undertaken to explore the interaction of envelope structural proteins with host molecules. However, how the nucleocapsid proteins of WSSV interacted with host molecules remained largely unknown. In this study, the interaction of nucleocapsid protein VP51 and ribosomal protein L7 of Litopenaeus vannamei (LvRPL7) was reported. Furthermore, the mRNA transcriptional response of LvRPL7 to WSSV was investigated. The results showed that LvRPL7 was widely distributed in all analyzed tissues of L. vannamei. The high expression levels of LvRPL7 were found in the tissues of muscle and gills. The temporal expression of LvRPL7 in WSSV-challenged shrimp showed that LvRPL7 was up-regulated (P WSSV challenge and then restored to the normal levels. But the LvRPL7 expression was up-regulated (P WSSV challenge and down-regulated at 12 h and 24 h post WSSV challenge. Indirect immunofluorescence assay indicated that LvRPL7 was mainly located on the surface and cytoplasm of hemocytes. Far-Western blotting showed that VP51 bound with LvRPL7. Moreover, ELISA results appeared that LvRPL7 interacted with VP51 in concentration dependent manner. Neutralization assay in vivo showed that anti-LvRPL7 antibody significantly delayed WSSV infection. Our results reveal that LvRPL7 was involved in WSSV infection. PMID:25736720

  16. 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.

    2006-01-01

    and reared for two weeks. Three repl i- cates were maintained for eac h feed. Mortality during this p e riod was observed to be negligible in all the tanks. Fibre reinforced tanks (30 l capacity) were used for the experiment. Water quality... . Therefore, after feeding on experime n tal diet for Table 1. Rearing conditions and water quality Initial body weight (average) 0.02166 g Stocking density 25 PL/tank Tank capacity 30 l Volum e of water 15 l Feeding level 15 ? 20...

  17. 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.

    2004-01-01

    and the mortality was also negligible (less than 5%). Histopathology of the treated shrimps also showed improvement in the health. The study also indicated the prophylactic property of the extract against WSSV infection. However, the study was carried out only under...

  18. Reação de híbridos de milho e comparação de métodos para avaliação da Cercosporiose e Mancha Branca / Reaction of commercial corn hybrids and comparison of methods for evaluation of maize White Spot and Gray Leaf Spot

    Scientific Electronic Library Online (English)

    André H. de, Brito; Renzo G. von, Pinho; Álvaro de O., Santos; Sidnei dos, Santos.

    2011-02-01

    Full Text Available O objetivo desse trabalho foi avaliar o nível de resistência de 12 híbridos comerciais de milho à cercosporiose, causada por Cercospora zeae-maydis, e à Mancha Branca, causada por Pantoea ananatis e comparar a eficiência das estimativas da área abaixo da curva de progresso das doenças e da estabilid [...] ade fenotípica, na avaliação da resistência a esses patógenos. Foram conduzidos três experimentos, no ano agrícola de 2007/2008, em três locais. Utilizou-se o delineamento em blocos ao acaso com três repetições. Foram realizadas cinco avaliações da severidade das doenças, em intervalos de sete dias, a partir dos 80 dias da emergência, por meio de uma escala de notas variando de 1 (altamente resistente) a 9 (altamente suscetível). Foram estimadas a área abaixo da curva de progresso da doença (AACPD) e o coeficiente de regressão linear (b1) entre a variável independente época de avaliação (x) e a variável dependente severidade da doença (y) e o coeficiente de determinação (R²). Observou-se que, ambas as metodologias utilizadas mostraram-se eficientes na discriminação do nível de resistência dos híbridos avaliados à Cercosporiose e à Mancha Branca, permitindo a classificação de modo semelhante. Os híbridos mais resistentes às duas doenças foram o AG7088, AG7010 e 2B707 e os mais suscetíveis o 30F44, 30F53 e AG8021. Os híbridos 30K64, DKB177, DKB390 e Impacto apresentaram comportamento variável em função dos locais de avaliação. Abstract in english With the objective of evaluating the resistance level of commercial corn hybrids and comparing the efficiency of the estimates of the area under the disease progress curve (AUDPC) and that of the phenotypical stability parameters in the evaluation of resistance maize white spot and gray leaf spot, t [...] hree experiments were carried out in the agricultural year of 2007/2008 in three environments. Twelve commercial maize hybrids were used, in a randomized block experimental design with three replications. Five evaluations of disease severity (maize white spot and gray leaf spot) based on visual symptoms were performed at seven-day intervals from the 80th day after maize emergence, ranging from 1 (highly resistant) to 9 (highly susceptible). The area under the disease progress curve (AUDPC) was estimated, as were the phenotypical stability parameters, i.e., the linear regression coefficient (b i) between the independent variable evaluation time (x), and the dependent variable, disease severity (y) and the determination coefficient (R²). It was found that in the case of maize white spot and maize gray leaf spot, both methodologies used proved to be effective in the discrimination of the resistance level of the hybrids, enabling them to be ranked in a similar way. The most resistant hybrids to maize white spot and maize gray leaf spot were AG7088, AG 7010 and 2B707, and the most susceptible were 30F44, 30F53 and AG8021. The 30K64, DKB177, DKB390 and Impacto hybrids showed variable levels of resistance with the environment.

  19. Thripinema fuscum (Tylenchida: Allantonematidae) parasitism reduces both the feeding of Frankliniella fusca (Thysanoptera: Thripidae) on peanut and the transmission of Tomato spotted wilt virus

    Science.gov (United States)

    Frankliniella fusca (Hinds) is the predominant thrips species found inhabiting and reproducing in peanut (Arachis hypogaea L.) and is one of at least nine thrips species to transmit Tomato spotted wilt virus (TSWV). The entomogenous nematode Thripinema fuscum Tipping & Nguyen, a natural enemy of F....

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

    Science.gov (United States)

    Nachappa, Punya; Margolies, David C; Nechols, James R; Whitfield, Anna E; Rotenberg, Dorith

    2013-01-01

    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. PMID:24058708

  1. Surveillance for West Nile virus in American white pelicans, Montana, USA, 2006-2007.

    Science.gov (United States)

    Johnson, Gregory; Nemeth, Nicole; Hale, Kristina; Lindsey, Nicole; Panella, Nicholas; Komar, Nicholas

    2010-03-01

    West Nile virus (WNV)-associated deaths of American white pelican (Pelecanus erythrorhynchos) chicks have been recognized at various nesting colonies in the United States since 2002. We evaluated American white pelican nesting colonies in Sheridan County, Montana, USA, for an association between WNV-positive pelican carcasses and human West Nile neuroinvasive disease. Persons in counties hosting affected colonies had a 5x higher risk for disease than those in counties with unaffected colonies. We also investigated WNV infection and blood meal source among mosquitoes and pelican tissue type for greatest WNV detection efficacy in carcasses. WNV-infected Culex tarsalis mosquitoes were detected and blood-engorged Cx. tarsalis contained pelican DNA. Viral loads and detection consistency among pelican tissues were greatest in feather pulp, brain, heart, and skin. Given the risks posed to wildlife and human health, coordinated efforts among wildlife and public health authorities to monitor these pelican colonies for WNV activity are potentially useful. PMID:20202414

  2. Hepatitis B virus core protein with hot-spot mutations inhibit MxA gene transcription but has no effect on inhibition of virus replication by interferon ?.

    Science.gov (United States)

    Zhijian, Yu; Zhen, Huang; Fan, Zhang; Jin, Yang; Qiwen, Deng; Zhongming, Zeng

    2010-01-01

    It has been reported that hepatitis B virus (HBV) core protein (HBc) can inhibit the transcription of human interferon-induced MxA gene. In this study, we investigated whether HBc protein mutations at hot spots (L60V, S87G and I97L) could still inhibit MxA transcription and the potential significance of this inhibition in virus replication in vitro. Our data indicated that the IFN-induced MxA mRNA expression level and MxA promoter activity was significantly down-regulated by mutant protein of HBc(I97L), compared to WT and the other two mutated HBc proteins(L60V or S87G). However, in Huh7 cells stably expressing WT or the mutated HBc proteins (L60V, S87G or I97L), IFN-? could inhibit the extra- and intracellular HBV DNA level and HBsAg secretion to a similar level compared to that in cells transfected with control plasmids. In conclusion, HBc protein with I97L mutation may play an special role in suppressing the transcription of MxA gene. Moreover, the inhibitory effect on MxA gene transcription by the WT or mutated HBc proteins (L60V, S87G and I97L) has no impact on inhibition of HBV replication by IFN-? in Huh7 cells. The clinical significance of the inhibitory effect of MxA gene transcription by HBc protein requires further study. PMID:20959021

  3. Pink spot, white spot: the pineal skylight of the leatherback turtle (Dermochelys coriacea Vandelli 1761) skull and its possible role in the phenology of feeding migrations

    Science.gov (United States)

    Davenport, John; Jones, T. Todd; Work, Thierry M.; Balazs, George H.

    2014-01-01

    Leatherback turtles, Dermochelys coriacea, which have an irregular pink area on the crown of the head known as the pineal or ‘pink spot’, forage upon jellyfish in cool temperate waters along the western and eastern margins of the North Atlantic during the summer. Our study showed that the skeletal structures underlying the pink spot in juvenile and adult turtles are compatible with the idea of a pineal dosimeter function that would support recognition of environmental light stimuli. We interrogated an extensive turtle sightings database to elucidate the phenology of leatherback foraging during summer months around Great Britain and Ireland and compared the sightings with historical data for sea surface temperatures and day lengths to assess whether sea surface temperature or light periodicity/levels were likely abiotic triggers prompting foraging turtles to turn south and leave their feeding grounds at the end of the summer. We found that sea temperature was too variable and slow changing in the study area to be useful as a trigger and suggest that shortening of day lengths as the late summer equilux is approached provides a credible phenological cue, acting via the pineal, for leatherbacks to leave their foraging areas whether they are feeding close to Nova Scotia or Great Britain and Ireland.

  4. Pathology of porcine peripheral white blood cells during infection with African swine fever virus

    Directory of Open Access Journals (Sweden)

    Karalyan Zaven

    2012-02-01

    Full Text Available Abstract Background African swine fever virus (ASFV is the causative agent of African swine fever (ASF that is the significant disease of domestic pigs. Several studies showed that ASFV can influence on porcine blood cells in vitro. Thus, we asked ourselves whether ASFV infection results in changes in porcine blood cells in vivo. A series of experiments were performed in order to investigate the effects of ASFV infection on porcine peripheral white blood cells. Nine pigs were inoculated by intramuscular injection with 104 50% hemadsorbing doses of virus (genotype II distributed in Armenia and Georgia. The total number of fifteen cell types was calculated during experimental infection. Results Although band-to-segmented neutrophils ratio became much higher (3.5 in infected pigs than in control group (0.3, marked neutropenia and lymphopenia were detected from 2 to 3 days post-infection. In addition to band neutrophils, the high number of other immature white blood cells, such as metamyelocytes, was observed during the course of infection. From the beginning of infection, atypical lymphocytes, with altered nuclear shape, arose and became 15% of total cells in the final phase of infection. Image scanning cytometry revealed hyperdiploid DNA content in atypical lymphocytes only from 5 days post-infection, indicating that DNA synthesis in pathological lymphocytes occurred in the later stages of infection. Conclusion From this study, it can be concluded that ASFV infection leads to serious changes in composition of white blood cells. Particularly, acute ASFV infection in vivo is accompanied with the emergence of immature cells and atypical lymphocytes in the host blood. The mechanisms underlying atypical cell formation remain to be elucidated.

  5. Febre maculosa das Montanhas Rochosas: ensaios negativos de transmissão experimental do virus por Triatomideos Rocky Mountain spotted fever: failure of Triatomid bugs to transmit the virus experimentally

    Directory of Open Access Journals (Sweden)

    Cornelius B. Philip

    1938-01-01

    Full Text Available 1. - As seguintes especies de Triatomideos não puderam transmitir pela picada o virus da febre maculosa das Montanhas Rochosas a cobayas normaes, nos seguintes prazos após a sucção de animal infectado: Eutriatoma uhleri, 33, 47, 75 e 141 dias (1 exemplar; Triatoma protracta, 15 e 37 dias (1 exemplar, Triatoma infestans, 8 dias (15 exemplares, e Rhodnius prolixus, 2 dias (1 exemplar. Foi demonstrado por inoculaçao que o ultimo insecto ainda continha o virus vivo. 2. - Experiencias de transmissão mechanica, por picada interrompida em animal infectado e continuada immediatamente em animal são, foram tambem negativas, com as especies T. protracta e R. prolixus. Um unico insecto da primeira especie picou 2 vezes cada animal, emquanto que 22 exemplares da segunda especie picaram de 1 a 3 vezes cada cobaya. 3. - A inoculação de gottas de dejecções de um R. prolixus eliminadas 2 dias depois de sugar animal infectado, não produziu a infecção em cobaya normal, não obstante ter sido demonstrada a presença do virus no organismo do barbeiro, por inoculação do conteúdo estomacal em outra cobaya. 4. - Foram feitas experiencias para determinar o tempo de sobrevivencia do virus nos barbeiros, inoculando-se conteúdo intestinal a diversos intervallos depois da sucção de cobayas infectadas, com os seguintes resultados: T. infestans: positivo 1 vez em 24 horas e 2 vezes em 48 horas; negativo 2 vezes em 72. 96, 120 e 192 horas. P. megistus: positivo 3 vezes em 24 horas, 2 vezes em 48 horas e 1 vez em 72 horas; negativo 1 vez em 72 e 96 horas; resultado duvidoso ou sem valor (infecção intercorrente 1 vez em 48 horas e 2 vezes a 72, 96 e 144 horas cada um. R. prolixus: positivo 1 vez em 24, 48 e 72 horas e negativo em 96 horas. 5. - Em vista dos resultados destas experiencias, feitas com 5 especies pertecentes a 4 generos de Triatomideos, torna-se muito pouco provavel que estes Hemipteros possam transmitir pela picada o virus da febre maculosa das Montanhas Rochosas, ou retel-o em seu organismo, em estado virulento, por mais de 2 a 4 dias.1. - The following species of blood-sucking triatomids failed to transmit the virus of Rocky Mountain spotted fever to susceptible guinea pigs by feeding at the following respective time intervals after the infective feeding: Eutriatoma uhleri, 33, 47, 75, and 141 days (one bug; Triatoma protracta, 15 and 37 days (one bug; T. infestans, 8 days (15 bugs; and Rhodnius prolixus, 2 days (1 bug. The last was shown to contain virus. 2. - Mechanical transmission tests by undelayed, interrupted feedings of 3 species, T. protracta and R. prolixus, were also negative. One insect of the former species accepted 2 infective and 2 normal (test feedings, while 22 bugs of the latter species accepted alternate blood-meals one to 3 times each on infected and normal guinea pigs. 3. - Fecal droplets collected from one R. prolixus 2 days after an infected feeding failed to infect when injected into a susceptible guinea pig, although virus was shown to be present inthe bug by subsequent injection of the viscera into another test animal. 4. - The period of survival of the virus in the bugs was determined by injection of gut contents at various short intervals after infected feedings. T. infestans: Positive once a 24 hours and twice at 48 hours; negative twice at 72, 96, 120 and 192 hours each. Panstrongylus megistus: Positive 3 times at 24 hours, twice at 48 hours, and once at 72 hours; negative once each at 72 and 96 hours; tests doubtful or valueless once at 48 hours, and twice each at 72, 96 and 144 hours. R. prolixus: Positive once each at 24, 48 and 72 hours, and negative at 96 hours. 5. - From these data, involving species of 4 genera of the Triatomidae, it appears unlikely that triatomids can either transmit Rocky Mountain spotted fever by their bites, or retain virulent virus within their bodies for longer than 2 to 4 days.

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

    OpenAIRE

    Ma Hao; Song Congfeng; Borth Wayne; Sether Diane; Melzer Michael; Hu John

    2011-01-01

    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 petun...

  7. Engineering cherry rootstocks with resistance to Prunus necrotic ring spot virus through RNAi-mediated silencing.

    Science.gov (United States)

    Song, Guo-qing; Sink, Kenneth C; Walworth, Aaron E; Cook, Meridith A; Allison, Richard F; Lang, Gregory A

    2013-08-01

    Prunus necrotic ringspot virus (PNRSV) is a major pollen-disseminated ilarvirus that adversely affects many Prunus species. In this study, an RNA interference (RNAi) vector pART27-PNRSV containing an inverted repeat (IR) region of PNRSV was transformed into two hybrid (triploid) cherry rootstocks, 'Gisela 6' (GI 148-1) and 'Gisela 7'(GI 148-8)', which are tolerant and sensitive, respectively, to PNRSV infection. One year after inoculation with PNRSV plus Prune Dwarf Virus, nontransgenic 'Gisela 6' exhibited no symptoms but a significant PNRSV titre, while the transgenic 'Gisela 6' had no symptoms and minimal PNRSV titre. The nontransgenic 'Gisela 7' trees died, while the transgenic 'Gisela 7' trees survived. These results demonstrate the RNAi strategy is useful for developing viral resistance in fruit rootstocks, and such transgenic rootstocks may have potential to enhance production of standard, nongenetically modified fruit varieties while avoiding concerns about transgene flow and exogenous protein production that are inherent for transformed fruiting genotypes. PMID:23521804

  8. 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.

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

    Directory of Open Access Journals (Sweden)

    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.

  10. Sintomas atípicos em frutos de meloeiro e de melancia ocasionados por Watermelon mosaic virus-2

    Directory of Open Access Journals (Sweden)

    LIMA J. ALBÉRSIO A.

    2002-01-01

    Full Text Available The Northeast of Brazil presents great potencial for melon (Cucumis melo and watermelon (Citrullus lanatus production. Melon fruit with white spots and watermelon fruits showing chlorotic spots were demonstrated to be caused by Watermelon mosaic virus-2, through indirect enzyme linked-immuno sorbent assay.

  11. Reconstructing the History of Maize Streak Virus Strain A Dispersal To Reveal Diversification Hot Spots and Its Origin in Southern Africa ? †

    OpenAIRE

    Monjane, Ade?rito L.; Harkins, Gordon W.; Martin, Darren P.; Lemey, Philippe; Lefeuvre, Pierre; Shepherd, Dionne N.; Oluwafemi, Sunday; Simuyandi, Michelo; Zinga, Innocent; Komba, Ephrem K.; Lakoutene, Didier P.; Mandakombo, Noella; Mboukoulida, Joseph; Semballa, Silla; Tagne, Appolinaire

    2011-01-01

    Maize streak virus strain A (MSV-A), the causal agent of maize streak disease, is today one of the most serious biotic threats to African food security. Determining where MSV-A originated and how it spread transcontinentally could yield valuable insights into its historical emergence as a crop pathogen. Similarly, determining where the major extant MSV-A lineages arose could identify geographical hot spots of MSV evolution. Here, we use model-based phylogeographic analyses of 353 fully sequen...

  12. Hepatitis B virus core protein with hot-spot mutations inhibit MxA gene transcription but has no effect on inhibition of virus replication by interferon ?

    Directory of Open Access Journals (Sweden)

    Qiwen Deng

    2010-10-01

    Full Text Available Abstract It has been reported that hepatitis B virus (HBV core protein (HBc can inhibit the transcription of human interferon-induced MxA gene. In this study, we investigated whether HBc protein mutations at hot spots (L60V, S87G and I97L could still inhibit MxA transcription and the potential significance of this inhibition in virus replication in vitro. Our data indicated that the IFN-induced MxA mRNA expression level and MxA promoter activity was significantly down-regulated by mutant protein of HBc(I97L, compared to WT and the other two mutated HBc proteins(L60V or S87G. However, in Huh7 cells stably expressing WT or the mutated HBc proteins (L60V, S87G or I97L, IFN-? could inhibit the extra- and intracellular HBV DNA level and HBsAg secretion to a similar level compared to that in cells transfected with control plasmids. In conclusion, HBc protein with I97L mutation may play an especial role in suppressing the transcription of MxA gene. Moreover, the inhibitory effect on MxA gene transcription by the WT or mutated HBc proteins (L60V, S87G and I97L has no impact on inhibition of HBV replication by IFN-? in Huh7 cells. The clinical significance of the inhibitory effect of MxA gene transcription by HBc protein requires further study.

  13. Parâmetros de cultivo e a enfermidade da mancha-branca em fazendas de camarões de Santa Catarina Cultivation parameters and the white spot disease in shrimp farms in Santa Catarina, Brazil

    Directory of Open Access Journals (Sweden)

    Sérgio Winckler da Costa

    2010-12-01

    Full Text Available O objetivo deste trabalho foi avaliar a relação entre parâmetros físico-químicos e biológicos do cultivo de camarão com a enfermidade causada pelo vírus da síndrome da mancha-branca (WSSV. Foram avaliadas a mortalidade, as características físicas e químicas da água e do solo, e foram realizados os exames e as análises microscópicas a fresco, presença de víbrios na hemolinfa, além do diagnóstico histopatológico e molecular (PCR do WSSV, em viveiros de oito fazendas de Santa Catarina, entre 2008 e 2009. O vírus foi detectado em cinco fazendas com registros da enfermidade em ciclos anteriores. A temperatura da água foi semelhante entre as fazendas com e sem WSSV, e a mortalidade ocorreu a intervalos ascendentes entre 24,6 e 29,3ºC. Não foram encontradas diferenças significativas nos parâmetros de análises microscópicas a fresco, tempo de coagulação e presença de víbrios na hemolinfa e sinais clínicos, entre viveiros com e sem WSSV. As concentrações de nitrito, sílica, fenol e alcalinidade na água e pH do solo apresentaram diferenças significativas entre os viveiros afetados ou não por WSSV. Os valores de nitrito, sílica e pH do solo estavam dentro dos limites recomendados para o cultivo de camarões, no entanto, isto não ocorreu com a alcalinidade e a concentração de fenol, o que sugere uma relação dos últimos com a manifestação da enfermidade da mancha-branca.The objective of this work was to evaluate the relationship among physicochemical and biological parameters in shrimp cultivation with the disease caused by the white spot syndrome virus (WSSV. The occurrence of mortality, physical and chemical characteristics of water and soil were evaluated, and clinical and microscopic analyses and the presence of vibrio in hemolymph, were performed as well as the histopathological and molecular diagnosis (PCR of the WSSV in ponds of eight farms in Santa Catarina, Brazil, between 2008 and 2009. The virus was detected in five farms, which had displayed the disease in previous cycles. The temperature of the water was similar among farms independently of the presence of the virus, and mortality occurred at ascendant temperature intervals, between 24.6 and 29.3ºC. No significant differences were found for the parameters of microscopic analyses among ponds either with or without the virus. Nitrite, silica, phenol and water-alkalinity concentrations, as well as soil pH values, showed significant differences among the ponds affected or not by the WSSV. Nitrite, silica and pH values were within the recommended limits for shrimp cultivation, however this did not occur for the alkalinity and phenol concentrations, wich suggests a relationship of the latter parameters with manifestation of the white spot disease.

  14. Parâmetros de cultivo e a enfermidade da mancha-branca em fazendas de camarões de Santa Catarina / Cultivation parameters and the white spot disease in shrimp farms in Santa Catarina, Brazil

    Scientific Electronic Library Online (English)

    Sérgio Winckler da, Costa; Luiz Rodrigo Mota, Vicente; Talita Medeiros de, Souza; Edemar Roberto, Andreatta; Maria Risoleta Freire, Marques.

    2010-12-01

    Full Text Available O objetivo deste trabalho foi avaliar a relação entre parâmetros físico-químicos e biológicos do cultivo de camarão com a enfermidade causada pelo vírus da síndrome da mancha-branca (WSSV). Foram avaliadas a mortalidade, as características físicas e químicas da água e do solo, e foram realizados os [...] exames e as análises microscópicas a fresco, presença de víbrios na hemolinfa, além do diagnóstico histopatológico e molecular (PCR) do WSSV, em viveiros de oito fazendas de Santa Catarina, entre 2008 e 2009. O vírus foi detectado em cinco fazendas com registros da enfermidade em ciclos anteriores. A temperatura da água foi semelhante entre as fazendas com e sem WSSV, e a mortalidade ocorreu a intervalos ascendentes entre 24,6 e 29,3ºC. Não foram encontradas diferenças significativas nos parâmetros de análises microscópicas a fresco, tempo de coagulação e presença de víbrios na hemolinfa e sinais clínicos, entre viveiros com e sem WSSV. As concentrações de nitrito, sílica, fenol e alcalinidade na água e pH do solo apresentaram diferenças significativas entre os viveiros afetados ou não por WSSV. Os valores de nitrito, sílica e pH do solo estavam dentro dos limites recomendados para o cultivo de camarões, no entanto, isto não ocorreu com a alcalinidade e a concentração de fenol, o que sugere uma relação dos últimos com a manifestação da enfermidade da mancha-branca. Abstract in english The objective of this work was to evaluate the relationship among physicochemical and biological parameters in shrimp cultivation with the disease caused by the white spot syndrome virus (WSSV). The occurrence of mortality, physical and chemical characteristics of water and soil were evaluated, and [...] clinical and microscopic analyses and the presence of vibrio in hemolymph, were performed as well as the histopathological and molecular diagnosis (PCR) of the WSSV in ponds of eight farms in Santa Catarina, Brazil, between 2008 and 2009. The virus was detected in five farms, which had displayed the disease in previous cycles. The temperature of the water was similar among farms independently of the presence of the virus, and mortality occurred at ascendant temperature intervals, between 24.6 and 29.3ºC. No significant differences were found for the parameters of microscopic analyses among ponds either with or without the virus. Nitrite, silica, phenol and water-alkalinity concentrations, as well as soil pH values, showed significant differences among the ponds affected or not by the WSSV. Nitrite, silica and pH values were within the recommended limits for shrimp cultivation, however this did not occur for the alkalinity and phenol concentrations, wich suggests a relationship of the latter parameters with manifestation of the white spot disease.

  15. RNAi-based inhibition of infectious myonecrosis virus replication in Pacific white shrimp Litopenaeus vannamei.

    Science.gov (United States)

    Feijó, Rubens Galdino; Maggioni, Rodrigo; Martins, Pedro Carlos; de Abreu, Keuly Ladislau; Oliveira-Neto, João Mafaldo; Guertler, Cristhiane; Justino, Emily Bruna; Perazzolo, Luciane Maria; Marins, Luis Fernando

    2015-05-21

    Disease in Pacific white shrimp Litopenaeus vannamei caused by the infectious myonecrosis virus (IMNV) causes significant socioeconomic impacts in infection-prone shrimp aquaculture regions. The use of synthetic dsRNA to activate an RNA interference (RNAi) response is being explored as a means of disease prophylaxis in farmed shrimp. Here, survival was tracked in L. vannamei injected with long synthetic dsRNAs targeted to IMNV open reading frame (ORF) 1a, ORF1b, and ORF2 genome regions prior to injection challenge with IMNV, and real-time RT-PCR was used to track the progress of IMNV infection and mRNA expression levels of the host genes sid1, dicer2, and argonaute2. Injection of dsRNAs targeting the ORF1a and ORF1b genes but not the ORF2 gene strongly inhibited IMNV replication over a 3 wk period following IMNV challenge, and resulted in 90 and 83% shrimp survival, respectively. Host gene mRNA expression data indicated that the Sid1 protein, which forms a transmembrane channel involved in cellular import/export of dsRNA, increased in abundance most significantly in shrimp groups that were most highly protected by virus-specific dsRNA injection. Subclinical IMNV infections present in the experimental L. vannamei used increased markedly in the 2 d between injection of any of the 4 virus-specific or non-specific dsRNAs tested and IMNV challenge. While handling and injection stress are implicated in increasing IMNV replication levels, the underlying molecular factors that may have been involved remain to be elucidated. PMID:25993884

  16. 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

    OpenAIRE

    Garci?a-marcos, Alberto; Pacheco, Remedios; Manzano, Aranzazu; Aguilar, Emmanuel; Tenllado, , Francisco

    2013-01-01

    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 necros...

  17. Hot spot computational identification: Application to the complex formed between the hen egg white lysozyme (HEL) and the antibody HyHEL-10

    Science.gov (United States)

    Moreira, I. S.; Fernandes, P. A.; Ramos, M. J.

    The definition and comprehension of the hot spots in an interface is a subject of primary interest for a variety of fields, including structure-based drug design. Therefore, to achieve an alanine mutagenesis computational approach that is at the same time accurate and predictive, capable of reproducing the experimental mutagenesis values is a major challenge in the computational biochemistry field. Antibody/protein antigen complexes provide one of the greatest models to study protein-protein recognition process because they have three fundamentally features: specificity, high complementary association and a small epitope restricted to the diminutive complementary determining regions (CDR) region, while the remainder of the antibody is largely invariant. Thus, we apply a computational mutational methodological approach to the study of the antigen-antibody complex formed between the hen egg white lysozyme (HEL) and the antibody HyHEL-10. A critical evaluation that focuses essentially on the limitations and advantages between different computational methods for hot spot determination, as well as between experimental and computational methodological approaches, is presented.

  18. Southern rice black-streaked dwarf virus: a white-backed planthopper transmitted fijivirus threadening rice production in Asia

    Directory of Open Access Journals (Sweden)

    GuohuiZhou

    2013-09-01

    Full Text Available Southern rice black-streaked dwarf virus (SRBSDV, a nonenveloped icosahedral virus with a genome of 10 double-stranded RNA segments, is a novel species in the genus Fijivirus (family Reoviridae first recognized in 2008. Rice plants infected with this virus exhibit symptoms similar to those caused by Rice black-streaked dwarf virus. Since 2009, the virus has rapidly spread and caused serious rice losses in East and Southeast Asia. Significant progress has been made in recent years in understanding this disease, especially about the functions of the viral genes, rice–virus–insect interactions, and epidemiology and control measures. The virus can be efficiently transmitted by the white-backed planthopper (WBPH, Sogatella furcifera in a persistent circulative propagative manner but cannot be transmitted by the brown planthopper (Nilaparvata lugens and small brown planthopper (Laodelphax striatellus. Rice, maize, Chinese sorghum (Coix lacryma-jobi and other grass weeds can be infected via WBPH. However, only rice plays a major role in the virus infection cycle because of the vector's preference. In Southeast Asia, WBPH is a long-distance migratory rice pest. The disease cycle can be described as follows: SRBSDV and its WBPH vector overwinter in warm tropical or sub-tropical areas; viruliferous WBPH adults carry the virus from south to north via long-distance migration in early spring, transmit the virus to rice seedlings in the newly colonized areas, and lay eggs on the infected seedlings; the next generation of WBPHs propagate on infected seedlings, become viruliferous, disperse, and cause new disease outbreaks. Several molecular and serological methods have been developed to detect SRBSDV in plant tissues and individual insects. Control measures based on protection from WBPH, including seedbed coverage, chemical seed treatments, and chemical spraying of seedlings, have proven effective in China.

  19. Resistencia genética de híbridos de tomate (Solanum lycopersicum L. (Mill.) Al virus del bronceado (TSWV) / Genetic resistance of tomato hybrids (Solanum lycopersicum L. (Mill.) to tomato spotted wilt virus (TSWV)

    Scientific Electronic Library Online (English)

    Julio, Gabriel; Daniel, Sanabria; Silene, Veramendi; Giovanna, Plata; Ada, Angulo; Mario, Crespo.

    2013-06-01

    Full Text Available La presente investigación se realizó en el invernadero y laboratorio de la Fundación PROINPA en Cochabamba - Bolivia en el 2012. El objetivo fue evaluar la resistencia y suscep-tibilidad de plantas a los virus Tomato spotted wilt virus - TSWV, Tomato cholorotic spot virus - TCSV y Groundnut ringspot [...] virus - GRSV en 10 híbridos de tomate mediante evaluación feno-típica y del patrón molecular (marcador SCAR Sw- 421), que distingue los homocigotos y hete-rocigotos resistentes del susceptible. Los resul-tados mostraron que el marcador SW-421 se co-localizó con el gen Sw-5 de resistencia a TSWV. Se observó la presencia de la banda de resistencia (R) para TSWV a 940 bp en las variedades PROINPA 2 (Aguaí) y PROINPA 9 (Bonita) en estado homocigoto dominante (Sw-5/Sw-5). Las variedades PROINPA 1 (Andinita), PROINPA 3 (Arami), PROINPA 4 (Yara), PROINPA 5 (Pintona), PROINPA 6 (Jasuka), y PROINPA 10 (Bola Pera), mostraron la banda resistencia (H) a TSWV a 900-940 bp en estado heterocigoto (Sw-5/Sw-5+). Solamente la variedad PROINPA 7 (Redonda), el padre 71 89S LACHING SW-5 y la variedad Shannon mostraron el gen de suscep-tibilidad (S) al TSWV a 900 bp en estado homo-cigoto recesivo (Sw-5+/Sw-5+). Los análisis de severidad y de DAS-ELISA fueron confirmados con el análisis molecular. Abstract in english This research was conducted at the PROINPA Foundation’s greenhouse and laboratory in Cochabamba, Bolivia in 2012. Its objective was to evaluate the resistance and susceptibility to Tomato spotted wilt virus - TSWV, Tomato cholorotic spot virus - TCSV and Groundnut ringspot virus - GRSV in 10 tomato [...] hybrids. Phenotypic and molecular pattern (SCAR marker SW-421) evaluations were performed in order to differentiate homozygous and heterozygous resistant from susceptible plants. Results showed that molecular marker Sw421 is co -located with the TSWV-resistance Sw-5 gene. A TSWV-resistance band (R) was observed at 940 bp and showed the homozygous presence of the Sw-5 allele (Sw-5/Sw-5) in PROINPA 2 (Aguai) and PROINPA 9 (Bonita) varieties. PROINPA 1 (Andinita), PROINPA 3 (Arami), PROINPA 4 (Yara), PROINPA 5 (Pintona), PROINPA 6 (Jasuka) and PROINPA 10 (Bola Pera) varieties, showed a TSWV resistance band (H) at 900-940 bp in the heterozygous state (Sw-5/Sw-5+). Only PROINPA 7 (Redonda), the male parent 71 LACHING 89S Sw-5 and the variety Shannon showed TSWV susceptibility gene (S) at 900 bp in the homozygous-recessive state (Sw-5+/Sw-5+). The results of the severity analysis and of DAS-ELISA were confirmed by the molecular analysis.

  20. Serological Evidence for Eastern Equine Encephalitis Virus Activity in White-Tailed Deer, Odocoileus virginianus, in Vermont, 2010

    OpenAIRE

    Berl, Erica; Eisen, Rebecca J.; Macmillan, Katherine; Swope, Bethany N.; Saxton-shaw, Kali D.; Graham, Alan C.; Turmel, Jon P.; Mutebi, John-paul

    2013-01-01

    Serum samples from 489 free-ranging white-tailed deer (Odocoileus virginianus) were screened for antibodies against the Eastern equine encephalitis virus (EEEV) using plaque reduction neutralization tests (PRNTs). EEEV antibodies were detected in 10.2% of serum samples. This is the first evidence that EEEV is present in Vermont. Serum was collected from deer in all 14 counties in the state, and positive EEEV sera were found in 12 (85%) of 14 counties, suggesting statewide EEEV activity in Ver...

  1. Manchas por alimentación de Frankliniella occidentalis (Thysanoptera: Thripidae) en uva de mesa blanca / Ring spots by feeding of Frankliniella occidentalis (Thysanoptera: Thripidae) on white table grapes

    Scientific Electronic Library Online (English)

    JAIME E., ARAYA; CARLOS, MERINO; FELIPE, SANTIBÁÑEZ; LUIS, SAZO.

    2014-06-01

    Full Text Available Entre 2006-2008 se hicieron siete ensayos en el Valle Central de Chile para verificar que la alimentación de Frankliniella occidentalis, después de la floración, causa manchas en uva de mesa blanca con pérdidas de hasta 50% en algunas áreas. Las bayas se protegieron con spinosad a 5,76 g ia/hL, apli [...] cado por inmersión de los racimos o por aspersión, cubriendo diversos períodos del desarrollo desde el cierre de racimos y terminaron en cosecha temprana. El daño a la cosecha alcanzó 70-99% en el control sin tratar y sólo 1-6% en los racimos que se protegieron de la alimentación del trips durante el cierre. Las aplicaciones desde la pinta de las bayas presentaron 22-55% de incidencia en los racimos, y aquellas temprano en la cosecha tuvieron daño similar al control. En todos los ensayos, los resultados de proteger los racimos contra los trips sólo al cierre fueron similares a los de aplicaciones múltiples en todos los estados fenológicos. En un ensayo en 2007-2008, una aplicación durante el cierre de racimos redujo significativamente el daño. Sin embargo, de los seis sitios seleccionados en 2006 por presentar las manchas en temporadas previas, sólo tres tuvieron daño significativo en el control sin tratar. La aparición variable de estas manchas debería inducir a los agricultores a evaluar la densidad de trips temprano en el cierre de racimos o hacer aplicaciones por calendario en este período para evitar daños. Abstract in english Seven trials were conducted during 2006-2008 in the Central Valley of Chile to verify that Frankliniella occidentalis feeding after flowering induce ring spots on white table grapes shortly before harvest, resulting in economic losses up to 50% in some areas. Treatment of spinosad at 5.76 g ai/hL we [...] re applied by dipping grape bunches or by spraying, across a range of fruit growth stages including bunch closure and veraison, and were stopped at early harvest. Ring spot incidence in bunches at harvest reached 70-99% in the untreated control compared to 1-6% in treatments where bunch closure was treated and protected from thrips. Applications starting at veraison resulted in 22- 55% incidence in bunches, while those started at early harvest time were equal in damage to the control. In all trials, results achieved by protecting against thrips feeding only at bunch closure were equal to multiple applications at all fruit growth stages. Damage levels in another trial during 2007-2008 for thrips control during bunch closure found that a single application at this stage can provide commercial ring spot russet reduction. However, of the six sites selected in 2006 due to ring spot russet damage previously, only three had significant damage in the untreated control. The variable appearance of ring spot russet should prompt growers to monitor thrip populations in bunches at early bunch closure or to do calendar sprays during this stage to avoid damage.

  2. Reconstructing the history of maize streak virus strain a dispersal to reveal diversification hot spots and its origin in southern Africa.

    Science.gov (United States)

    Monjane, Adérito L; Harkins, Gordon W; Martin, Darren P; Lemey, Philippe; Lefeuvre, Pierre; Shepherd, Dionne N; Oluwafemi, Sunday; Simuyandi, Michelo; Zinga, Innocent; Komba, Ephrem K; Lakoutene, Didier P; Mandakombo, Noella; Mboukoulida, Joseph; Semballa, Silla; Tagne, Appolinaire; Tiendrébéogo, Fidèle; Erdmann, Julia B; van Antwerpen, Tania; Owor, Betty E; Flett, Bradley; Ramusi, Moses; Windram, Oliver P; Syed, Rizwan; Lett, Jean-Michel; Briddon, Rob W; Markham, Peter G; Rybicki, Edward P; Varsani, Arvind

    2011-09-01

    Maize streak virus strain A (MSV-A), the causal agent of maize streak disease, is today one of the most serious biotic threats to African food security. Determining where MSV-A originated and how it spread transcontinentally could yield valuable insights into its historical emergence as a crop pathogen. Similarly, determining where the major extant MSV-A lineages arose could identify geographical hot spots of MSV evolution. Here, we use model-based phylogeographic analyses of 353 fully sequenced MSV-A isolates to reconstruct a plausible history of MSV-A movements over the past 150 years. We show that since the probable emergence of MSV-A in southern Africa around 1863, the virus spread transcontinentally at an average rate of 32.5 km/year (95% highest probability density interval, 15.6 to 51.6 km/year). Using distinctive patterns of nucleotide variation caused by 20 unique intra-MSV-A recombination events, we tentatively classified the MSV-A isolates into 24 easily discernible lineages. Despite many of these lineages displaying distinct geographical distributions, it is apparent that almost all have emerged within the past 4 decades from either southern or east-central Africa. Collectively, our results suggest that regular analysis of MSV-A genomes within these diversification hot spots could be used to monitor the emergence of future MSV-A lineages that could affect maize cultivation in Africa. PMID:21715477

  3. Reconstructing the History of Maize Streak Virus Strain A Dispersal To Reveal Diversification Hot Spots and Its Origin in Southern Africa ? †

    Science.gov (United States)

    Monjane, Adérito L.; Harkins, Gordon W.; Martin, Darren P.; Lemey, Philippe; Lefeuvre, Pierre; Shepherd, Dionne N.; Oluwafemi, Sunday; Simuyandi, Michelo; Zinga, Innocent; Komba, Ephrem K.; Lakoutene, Didier P.; Mandakombo, Noella; Mboukoulida, Joseph; Semballa, Silla; Tagne, Appolinaire; Tiendrébéogo, Fidèle; Erdmann, Julia B.; van Antwerpen, Tania; Owor, Betty E.; Flett, Bradley; Ramusi, Moses; Windram, Oliver P.; Syed, Rizwan; Lett, Jean-Michel; Briddon, Rob W.; Markham, Peter G.; Rybicki, Edward P.; Varsani, Arvind

    2011-01-01

    Maize streak virus strain A (MSV-A), the causal agent of maize streak disease, is today one of the most serious biotic threats to African food security. Determining where MSV-A originated and how it spread transcontinentally could yield valuable insights into its historical emergence as a crop pathogen. Similarly, determining where the major extant MSV-A lineages arose could identify geographical hot spots of MSV evolution. Here, we use model-based phylogeographic analyses of 353 fully sequenced MSV-A isolates to reconstruct a plausible history of MSV-A movements over the past 150 years. We show that since the probable emergence of MSV-A in southern Africa around 1863, the virus spread transcontinentally at an average rate of 32.5 km/year (95% highest probability density interval, 15.6 to 51.6 km/year). Using distinctive patterns of nucleotide variation caused by 20 unique intra-MSV-A recombination events, we tentatively classified the MSV-A isolates into 24 easily discernible lineages. Despite many of these lineages displaying distinct geographical distributions, it is apparent that almost all have emerged within the past 4 decades from either southern or east-central Africa. Collectively, our results suggest that regular analysis of MSV-A genomes within these diversification hot spots could be used to monitor the emergence of future MSV-A lineages that could affect maize cultivation in Africa. PMID:21715477

  4. Selection of shrimp breeders free of white spot syndrome and infectious hypodermal and hematopoietic necrosis / Seleção de reprodutores de camarão livres da síndrome da mancha-branca e da necrose infecciosa hipodermal e hematopoiética

    Scientific Electronic Library Online (English)

    Carlos Cesar de, Mello Junior; Gael Yvan Leclercq, Delsol; Emmerik, Motte; Virna Alexia Cedeño, Escobar; Pedro Filipe, Rey; Mauricio Laterça, Martins; Luis Alejandro Vinatea, Arana; Giovanni Lemos de, Mello; Alvaro Pestana de, Farias; Xavier Antonio Serrano, Arguello; John Erick Montaño, Maridueña.

    2011-05-01

    Full Text Available O objetivo deste trabalho foi selecionar reprodutores de Litopenaeus vannamei sobreviventes de um surto do vírus da síndrome da mancha-branca (WSSV), adaptados às condições locais e diagnosticados negativamente para WSSV e para o vírus da necrose infecciosa hipodermal e hematopoiética (IHHNV), e ava [...] liar se esta extratégia é uma alternativa viável para produção em Santa Catarina. Foram selecionados fenotipicamente 800 machos e 800 fêmeas, de um viveiro. Análises de nested-PCR de 487 fêmeas e de 231 machos, sexualmente maduros, mostraram que 63% das fêmeas e 55% dos machos estavam infectados com IHHNV. Os animais livres de IHHNV foram testados para WSSV, e os considerados duplo negativos destinados à reprodução. As pós-larvas produzidas foram estocadas em nove berçários, para análise. Das 45 amostras, com 50 pós-larvas cada, apenas duas foram positivas para IHHNV e nenhuma para WSSV. Os lotes de pós-larvas diagnosticadas livres de vírus por nested-PCR foram encaminhados para seis fazendas. Foi realizada análise comparativa em viveiros de engorda, entre pós-larvas locais e pós-larvas do Nordeste do Brasil. Também foram analisados caranguejos (Chasmagnathus granulata), siris (Callinectes sapidus) e lebres do mar (Aplysia brasiliana), que são possíveis vetores dos vírus. A média de sobrevivência foi de 55% para as pós-larvas locais e de 23,4% para as pós-larvas do Nordeste. As lebres do mar apresentaram prevalência de 50% e os caranguejos de 67% de WSSV. Abstract in english The objective of this work was to select surviving breeders of Litopenaeus vannamei from white spot syndrome virus (WSSV) outbreak, adapted to local climatic conditions and negatively diagnosed for WSSV and infectious hypodermal and hematopoietic necrosis virus (IHHNV), and to evaluate if this strat [...] egy is a viable alternative for production in Santa Catarina, Brazil. A total of 800 males and 800 females were phenotypically selected in a farm pond. Nested-PCR analyses of 487 sexually mature females and 231 sexually mature males showed that 63% of the females and 55% of the males were infected with IHHNV. Animals free of IHHNV were tested for WSSV, and those considered double negative were used for breeding. The post-larvae produced were stocked in nine nursery tanks for analysis. From the 45 samples, with 50 post-larvae each, only two were positive for IHHNV and none for WSSV. Batches of larvae diagnosed free of virus by nested-PCR were sent to six farms. A comparative analysis was carried out in growth ponds, between local post-larvae and post-larvae from Northeast Brazil. Crabs (Chasmagnathus granulata), blue crabs (Callinectes sapidus), and sea hares (Aplysia brasiliana), which are possible vectors of these viruses, were also evaluated. The mean survival was 55% for local post-larvae against 23.4% for post-larvae from the Northeast. Sea hares showed prevalence of 50% and crabs of 67% of WSSV.

  5. Selection of shrimp breeders free of white spot syndrome and infectious hypodermal and hematopoietic necrosis Seleção de reprodutores de camarão livres da síndrome da mancha-branca e da necrose infecciosa hipodermal e hematopoiética

    Directory of Open Access Journals (Sweden)

    Carlos Cesar de Mello Junior

    2011-05-01

    Full Text Available The objective of this work was to select surviving breeders of Litopenaeus vannamei from white spot syndrome virus (WSSV outbreak, adapted to local climatic conditions and negatively diagnosed for WSSV and infectious hypodermal and hematopoietic necrosis virus (IHHNV, and to evaluate if this strategy is a viable alternative for production in Santa Catarina, Brazil. A total of 800 males and 800 females were phenotypically selected in a farm pond. Nested-PCR analyses of 487 sexually mature females and 231 sexually mature males showed that 63% of the females and 55% of the males were infected with IHHNV. Animals free of IHHNV were tested for WSSV, and those considered double negative were used for breeding. The post-larvae produced were stocked in nine nursery tanks for analysis. From the 45 samples, with 50 post-larvae each, only two were positive for IHHNV and none for WSSV. Batches of larvae diagnosed free of virus by nested-PCR were sent to six farms. A comparative analysis was carried out in growth ponds, between local post-larvae and post-larvae from Northeast Brazil. Crabs (Chasmagnathus granulata, blue crabs (Callinectes sapidus, and sea hares (Aplysia brasiliana, which are possible vectors of these viruses, were also evaluated. The mean survival was 55% for local post-larvae against 23.4% for post-larvae from the Northeast. Sea hares showed prevalence of 50% and crabs of 67% of WSSV.O objetivo deste trabalho foi selecionar reprodutores de Litopenaeus vannamei sobreviventes de um surto do vírus da síndrome da mancha-branca (WSSV, adaptados às condições locais e diagnosticados negativamente para WSSV e para o vírus da necrose infecciosa hipodermal e hematopoiética (IHHNV, e avaliar se esta extratégia é uma alternativa viável para produção em Santa Catarina. Foram selecionados fenotipicamente 800 machos e 800 fêmeas, de um viveiro. Análises de nested-PCR de 487 fêmeas e de 231 machos, sexualmente maduros, mostraram que 63% das fêmeas e 55% dos machos estavam infectados com IHHNV. Os animais livres de IHHNV foram testados para WSSV, e os considerados duplo negativos destinados à reprodução. As pós-larvas produzidas foram estocadas em nove berçários, para análise. Das 45 amostras, com 50 pós-larvas cada, apenas duas foram positivas para IHHNV e nenhuma para WSSV. Os lotes de pós-larvas diagnosticadas livres de vírus por nested-PCR foram encaminhados para seis fazendas. Foi realizada análise comparativa em viveiros de engorda, entre pós-larvas locais e pós-larvas do Nordeste do Brasil. Também foram analisados caranguejos (Chasmagnathus granulata, siris (Callinectes sapidus e lebres do mar (Aplysia brasiliana, que são possíveis vetores dos vírus. A média de sobrevivência foi de 55% para as pós-larvas locais e de 23,4% para as pós-larvas do Nordeste. As lebres do mar apresentaram prevalência de 50% e os caranguejos de 67% de WSSV.

  6. PRELIMINARY RESULTS OF THE STUDY ON THE SPREAD OF APPLE CHLOROTIC LEAF SPOT VIRUS (ACLSV IN DIFFERENT FRUIT TREE SPECIES IN KYUSTENDIL REGION OF BULGARIA

    Directory of Open Access Journals (Sweden)

    Aneliya BORISOVA

    2005-08-01

    Full Text Available The survey was carried out mostly in one of the main fruit tree growing regions of Bulgaria - Kyustendil, during the period of 2004-2005. A total of 632 trees corresponding to 50 apple, 27 pear, 19 plum, 4 peach, 9 sweet cherry and 4 sour cherry cultivars and 21 apricot elites were tested for the presence of Apple chlorotic leaf spot virus (ACLSV and six other viruses by ELISA. Samples for analyses were collected from different organs of plants: flowers, leaves or phloem tissues. In the present research, ACLSV was not found in pear, plum, apricot and sour cherry. The highest infection rate of the virus among the infected fruit tree species was in apple (73 % followed by sweet cherry (13.3 % and peach (11.76 %. The frequency of mixed infection was 43.2 % in the infected apple, 33.4 % in sweet cherry and 12.5 % in peach trees. The highest concentrations of ACLSV, measured by ELISA, were observed in naturally grown flower petals of apple and sweet cherry trees in May.

  7. Prohibitin Interacts with Envelope Proteins of White Spot Syndrome Virus and Prevents Infection in the Red Swamp Crayfish, Procambarus clarkii

    OpenAIRE

    Lan, Jiang-Feng; Li, Xin-Cang; Sun, Jie-Jie; GONG, JING; Wang, Xian-Wei; Shi, Xiu-Zhen; Shi, Li-Jie; Weng, Yu-Ding; Zhao, Xiao-Fan; Wang, Jin-Xing

    2013-01-01

    Prohibitins (PHBs) are ubiquitously expressed conserved proteins in eukaryotes that are associated with apoptosis, cancer formation, aging, stress responses, cell proliferation, and immune regulation. However, the function of PHBs in crustacean immunity remains largely unknown. In the present study, we identified a PHB in Procambarus clarkii red swamp crayfish, which was designated PcPHB1. PcPHB1 was widely distributed in several tissues, and its expression was significantly upregulated by wh...

  8. Identification of orange-spotted grouper (Epinephelus coioides) interferon regulatory factor 3 involved in antiviral immune response against fish RNA virus.

    Science.gov (United States)

    Huang, Youhua; Huang, Xiaohong; Cai, Jia; OuYang, Zhengliang; Wei, Shina; Wei, Jingguang; Qin, Qiwei

    2015-02-01

    Interferon regulatory factor 3 (IRF3) is an important transcription factor which regulates the expression of interferon (IFN) and IFN-stimulated genes (ISGs) following virus recognition. In this study, a novel IRF3 gene was cloned from grouper Epinephelus coioides (EcIRF3) and its effects against Singapore grouper iridovirus (SGIV) and red spotted grouper nervous necrosis virus (RGNNV) was investigated. The full-length of EcIRF3 cDNA was composed of 2513 bp and encoded a polypeptide of 458 amino acids which shared 82% identity with European seabass (Dicentrarchus labrax). EcIRF3 contained three conserved domains including a DNA-binding domain (DBD), an IRF associated domain (IAD) and a serine-rich domain. Expression profile analysis revealed that EcIRF3 was abundant in head kidney, kidney, spleen and gill. Upon different stimuli in vitro, the transcript of EcIRF3 was significantly up-regulated after RGNNV infection or treatment with polyinosin-polycytidylic acid (poly I:C). During SGIV infection, the increase of the EcIRF3 transcription was only detected at the late stage, suggesting that EcIRF3 was differently regulated by different stimuli. Immune fluorescence assay indicated that the fluorescence signal of EcIRF3 was increased significantly after infection with RGNNV or treatment with poly I:C, but moderately at the late stage of SGIV infection. Reporter gene assay showed that EcIRF3 activated zebrafish type I IFN and type III IFN promoter in vitro. The viral gene transcription and virus production of RGNNV were significantly decreased in EcIRF3 overexpressing cells. However, the ectopic expression of EcIRF3 did not affect the gene transcription and virus production of SGIV. Moreover, the mRNA expression levels of type I IFN and IFN-inducible genes (MxI, ISG15 and ISG56) were increased in RGNNV infected EcIRF3 overexpressing cells compared to empty vector transfected cells. Together, our results demonstrated that IFN immune response mediated by grouper IRF3 was exerted crucial roles for fish RNA virus, but not for DNA virus replication. PMID:25463297

  9. Analysis of resistance to Yam mosaic virus, genus Potyvirus in white guinea yam (Dioscorea rotundata Poir. genotypes

    Directory of Open Access Journals (Sweden)

    Babajide Odu O.

    2011-01-01

    Full Text Available Resistance to Yam mosaic virus (YMV, genus Potyvirus was studied in 10 populations of selected white Guinea yam (Dioscorea rotundata. Plants of resistant genotypes: TDr 35, TDr 1621, TDr 93-1, TDr 93-32, TDr 95-107, TDr 93-23, and susceptible ones: TDr 87/00211, TDr 87/00571 and TDr 95-127 were screened for their reaction to the pathogen by symptom severity scoring scale of 1-5, and by quantifying virus multiplication by triple antibody sandwich enzyme-linked immunosorbent assay (TAS-ELISA. Controlled crosses were made among the genotypes within and between the groups according to reactions to the pathogen. The resultant F1 progenies were evaluated for the infection by disease symptom development and by TAS ELISA to detect a symptomless infection in an insect-proof screenhouse for the assessment of inheritance of resistance to YMV. A genetic analysis of the reactions of progenies derived from the D. rotundata genotypes to inoculation with YMV strongly suggests that resistance to the virus is a dominantly inherited trait. Segregation ratios obtained from the families indicate that at least two dominant genes are involved.

  10. Cache Valley and Potosi viruses (Bunyaviridae) in white-tailed deer (Odocoileus virginianus): experimental infections and antibody prevalence in natural populations.

    Science.gov (United States)

    Blackmore, C G; Grimstad, P R

    1998-11-01

    Cache Valley virus (CVV) and Potosi virus (POTV) are two closely related mosquito-borne viruses (Bunyaviridae: Bunyamwera group) that appear to circulate in several regions of the United States, especially the Midwest. We determined the prevalence of specific neutralizing antibodies to both viruses in Indiana white-tailed deer and conducted infection experiments to assess whether deer could serve as an vertebrate-amplifying host. Cross-infection experiments also were carried out to investigate the level of antibody cross-reactivity and cross-protection between the two viruses. The seroprevalence rate was high for both CVV (> 66%) and POTV (> 43%) in adult deer statewide. Antibodies neutralizing CVV were more common among deer harvested in the northern part of Indiana whereas the prevalence of POTV antibodies suggested a more southern distribution for this virus. Experimental infections of captive deer showed that they may serve as amplifying hosts for either virus. Deer infected with CVV or POTV developed a 1-3-day viremia with 3.0 and 4.1 log10 plaque-forming units/ml mean peak titers, respectively. However, significant levels of antibody cross-reactivity between the two viruses were observed. Viremia was lower and shorter when animals immune to either CVV or POTV were cross-infected with the alternate virus and antibody responses following cross-infections resembled original antigenic sin with higher titers of antibodies against the primary agent. PMID:9840585

  11. Serological evidence for eastern equine encephalitis virus activity in white-tailed deer, Odocoileus virginianus, in Vermont, 2010.

    Science.gov (United States)

    Berl, Erica; Eisen, Rebecca J; MacMillan, Katherine; Swope, Bethany N; Saxton-Shaw, Kali D; Graham, Alan C; Turmel, Jon P; Mutebi, John-Paul

    2013-01-01

    Serum samples from 489 free-ranging white-tailed deer (Odocoileus virginianus) were screened for antibodies against the Eastern equine encephalitis virus (EEEV) using plaque reduction neutralization tests (PRNTs). EEEV antibodies were detected in 10.2% of serum samples. This is the first evidence that EEEV is present in Vermont. Serum was collected from deer in all 14 counties in the state, and positive EEEV sera were found in 12 (85%) of 14 counties, suggesting statewide EEEV activity in Vermont. Analysis of the spatial distribution of PRNT-positive samples revealed a random distribution of EEEV throughout the state. The results indicate widespread EEEV activity in Vermont and suggest that EEEV is not a recent introduction to the state but that EEEV activity has not been detected until now. PMID:23208886

  12. Experimental infection of white-tailed deer (Odocoileus virginianus) with Northern European bluetongue virus serotype 8

    Science.gov (United States)

    Bluetongue (BT) is an insect-transmitted, economically important disease of domestic and wild ruminants. Although only five of the 26 reported bluetongue virus (BTV) serotypes are considered endemic to the USA, 10 exotic serotypes have been isolated primarily in the southeastern region of the count...

  13. Correlation between Human Immunodeficiency Virus Type 1 (HIV-1) RNA Measurements Obtained with Dried Blood Spots and Those Obtained with Plasma by Use of Nuclisens EasyQ HIV-1 and Abbott RealTime HIV Load Tests ?

    OpenAIRE

    Garrido, Carolina; Zahonero, Natalia; Corral, Ange?lica; Arredondo, Miguel; Soriano, Vincent; Mendoza, Carmen

    2009-01-01

    The plasma human immunodeficiency virus (HIV) RNA load is used in the clinical routine for the monitoring of HIV infection and the patient's response to antiretroviral therapy. Other body fluids or dried blood spots (DBS) can be used, however, to assess the level of viremia. The use of DBS may be especially helpful for the monitoring of HIV-infected patients in resource-poor settings, where access to adequate laboratory facilities is often difficult. However, the correlation between the HIV R...

  14. Strategies to facilitate the development of uncloned or cloned infectious full-length viral cDNAs: Apple chlorotic leaf spot virus as a case study

    Directory of Open Access Journals (Sweden)

    Marais Armelle

    2011-10-01

    Full Text Available Abstract Background Approaches to simplify and streamline the construction of full-length infectious cDNA clones (FL-cDNAs are needed. Among desirable improvements are the ability to use total nucleic acids (TNA extracts from infected hosts (to bypass viral purification limitations for the direct one-step amplification of large FL-cDNAs, the possibility to inoculate plants with uncloned FL-cDNAs and the simplified cloning of these large molecules. Results Using the 7.55 kb genome of Apple chlorotic leaf spot trichovirus (ACLSV approaches allowing the rapid generation from TNA extracts of FL-cDNAs under the control of the T7 promoter and the successful inoculation of plants using in vitro transcripts obtained from these uncloned amplification products have been developed. We also show that the yeast homologous recombination system permits efficient cloning of FL-cDNAs and the simultaneous one-step tailoring of a ternary Yeast-Escherichia coli-Agrobacterium tumefaciens shuttle vector allowing efficient inoculation of both herbaceous and woody host plants by agroinfiltration. Conclusions The fast and efficient strategies described here should have broad applications, in particular for the study of "difficult" plant viruses, such as those infecting woody hosts, and potentially for other, non plant-infecting viral agents.

  15. Measurement of Kodaikanal White-Light Images: Relaxation of Tilts of Spot Groups as Indicator of Subsurface Dynamics of Parent Flux Loops

    Science.gov (United States)

    Sivaraman, K. R.; Gokhale, M. H.; Sivaraman, Hari; Gupta, S. S.; Howard, Robert F.

    2007-03-01

    We reexamine the evolution of the observed tilts ? of spot groups with life spans 2-7 days in the two latitude belts 13°. Using an iterative procedure, we refine the linear fit between ? and the daily tilt angle changes ?? and obtain reliable estimates of the fit coefficients. We interpret our results in light of the scenario implied by the theoretical model of Longcope & Choudhuri for the subsurface dynamics of parent flux loops of bipolar magnetic regions and arrive at the following conclusions: (1) the parent flux tubes of spot groups possess a nonzero tilt at the onset of rise from the depths of their origin; these ``inborn tilts'' are ~4°-11° in latitudes 13° (2) during the rise the tilt of the omega loops of spot groups living 2-7 days get reduced to ~2°-6° in both the latitude belts, and this calls for reexamination of the role of Coriolis force as understood so far; (3) after emergence of the top of the loop above the surface, magnetic tension in the legs tends to restore the tilt to the inborn tilt on timescales of ~5 to 14 days; and (4) these timescales correspond to field strengths in the range ~14-40 kG for the parent flux loops and are close to the limits set by Fan et al.

  16. Southern rice black-streaked dwarf virus: a white-backed planthopper-transmitted fijivirus threatening rice production in Asia

    OpenAIRE

    GuohuiZhou

    2013-01-01

    Southern rice black-streaked dwarf virus (SRBSDV), a nonenveloped icosahedral virus with a genome of 10 double-stranded RNA segments, is a novel species in the genus Fijivirus (family Reoviridae) first recognized in 2008. Rice plants infected with this virus exhibit symptoms similar to those caused by Rice black-streaked dwarf virus. Since 2009, the virus has rapidly spread and caused serious rice losses in East and Southeast Asia. Significant progress has been made in recent years in underst...

  17. Southern rice black-streaked dwarf virus: a white-backed planthopper transmitted fijivirus threadening rice production in Asia

    OpenAIRE

    GuohuiZhou

    2013-01-01

    Southern rice black-streaked dwarf virus (SRBSDV), a nonenveloped icosahedral virus with a genome of 10 double-stranded RNA segments, is a novel species in the genus Fijivirus (family Reoviridae) first recognized in 2008. Rice plants infected with this virus exhibit symptoms similar to those caused by Rice black-streaked dwarf virus. Since 2009, the virus has rapidly spread and caused serious rice losses in East and Southeast Asia. Significant progress has been made in recent years in underst...

  18. Tomato spotted wilt virus (TS WV), weeds and thrip vectors in the tomato (Solanum lycopersicum L.) in the Andean region of Cundinamarca (Colombia) / Tomato spotted wilt virus (TSWV), malezas y vectores de trips en el tomate (Solanum lycopersicum L.) en la región andina de Cundinamarca (Colombia)

    Scientific Electronic Library Online (English)

    Everth E, Ebratt R; Rocio, Acosta A; Olga Y, Martínez B; Omar, Guerrero G; Walther, Turizo A.

    2013-04-01

    Full Text Available La presencia y distribución de TSWV, arvenses y los trips vectores en las principales zonas productoras de tomate en la región andina del departamento de Cundinamarca (provincias de Oriente, Sumapaz y Ubaté), se confirmó mediante la técnica DAS ELISA , se evaluó la presencia del virus TSWV en tejido [...] de tomate, arvenses y trips asociados. Se observaron incidencias altas en diferentes provincias de la región andina del departamento de Cundinamarca. La incidencia viral alcanzó promedios de 23,3% en el Sumapaz, el 19,4% en Oriente y el 4% en Ubaté. Los síntomas característicos observados correspondieron a manchas de color café y anillos concéntricos en el área foliar, tallos y frutos, bronceado, manchas en flor y marchitez en hojas, tallos y flores. Las especies de trips con mayor presencia fueron Frankliniella occidentalis, seguida de Thrips palmi y Thrips tabaci. Se determinó el importante papel de las arvenses como fuente de inóculo y reservorio de vectores en las especies Emilia sonchifolia y Amaranthus dubius Abstract in english 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 w [...] eeds. 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

  19. Immunological-based assays for specific detection of shrimp viruses

    Science.gov (United States)

    Chaivisuthangkura, Parin; Longyant, Siwaporn; Sithigorngul, Paisarn

    2014-01-01

    Among shrimp viral pathogens, white spot syndrome virus (WSSV) and yellow head virus (YHV) are the most lethal agents, causing serious problems for both the whiteleg shrimp, Penaeus (Litopenaeus) vannamei, and the black tiger shrimp, Penaeus (Penaeus) monodon. Another important virus that infects P. vannamei is infectious myonecrosis virus (IMNV), which induces the white discoloration of affected muscle. In the cases of taura syndrome virus and Penaeus stylirostris densovirus (PstDNV; formerly known as infectious hypodermal and hematopoietic necrosis virus), their impacts were greatly diminished after the introduction of tolerant stocks of P. vannamei. Less important viruses are Penaeus monodon densovirus (PmDNV; formerly called hepatopancreatic parvovirus), and Penaeus monodon nucleopolyhedrovirus (PemoNPV; previously called monodon baculovirus). For freshwater prawn, Macrobrachium rosenbergii nodavirus and extra small virus are considered important viral pathogens. Monoclonal antibodies (MAbs) specific to the shrimp viruses described above have been generated and used as an alternative tool in various immunoassays such as enzyme-linked immunosorbent assay, dot blotting, Western blotting and immunohistochemistry. Some of these MAbs were further developed into immunochromatographic strip tests for the detection of WSSV, YHV, IMNV and PemoNPV and into a dual strip test for the simultaneous detection of WSSV/YHV. The strip test has the advantages of speed, as the result can be obtained within 15 min, and simplicity, as laboratory equipment and specialized skills are not required. Therefore, strip tests can be used by shrimp farmers for the pond-side monitoring of viral infection. PMID:24567913

  20. Immunological-based assays for specific detection of shrimp viruses.

    Science.gov (United States)

    Chaivisuthangkura, Parin; Longyant, Siwaporn; Sithigorngul, Paisarn

    2014-02-12

    Among shrimp viral pathogens, white spot syndrome virus (WSSV) and yellow head virus (YHV) are the most lethal agents, causing serious problems for both the whiteleg shrimp, Penaeus (Litopenaeus) vannamei, and the black tiger shrimp, Penaeus (Penaeus) monodon. Another important virus that infects P. vannamei is infectious myonecrosis virus (IMNV), which induces the white discoloration of affected muscle. In the cases of taura syndrome virus and Penaeus stylirostris densovirus (PstDNV; formerly known as infectious hypodermal and hematopoietic necrosis virus), their impacts were greatly diminished after the introduction of tolerant stocks of P. vannamei. Less important viruses are Penaeus monodon densovirus (PmDNV; formerly called hepatopancreatic parvovirus), and Penaeus monodon nucleopolyhedrovirus (PemoNPV; previously called monodon baculovirus). For freshwater prawn, Macrobrachium rosenbergii nodavirus and extra small virus are considered important viral pathogens. Monoclonal antibodies (MAbs) specific to the shrimp viruses described above have been generated and used as an alternative tool in various immunoassays such as enzyme-linked immunosorbent assay, dot blotting, Western blotting and immunohistochemistry. Some of these MAbs were further developed into immunochromatographic strip tests for the detection of WSSV, YHV, IMNV and PemoNPV and into a dual strip test for the simultaneous detection of WSSV/YHV. The strip test has the advantages of speed, as the result can be obtained within 15 min, and simplicity, as laboratory equipment and specialized skills are not required. Therefore, strip tests can be used by shrimp farmers for the pond-side monitoring of viral infection. PMID:24567913

  1. Performance of a Novel Human Immunodeficiency Virus (HIV) Type 1 Total Nucleic Acid-Based Real-Time PCR Assay Using Whole Blood and Dried Blood Spots for Diagnosis of HIV in Infants?

    OpenAIRE

    Stevens, Wendy; Erasmus, Linda; Moloi, Matsidisho; Taleng, Thabo; Sarang, Somaya

    2008-01-01

    The new Cobas AmpliPrep/Cobas TaqMan HIV-1 Qual test offers advanced automation for the detection of human immunodeficiency virus type 1 (HIV-1) RNA and DNA in dried blood spots (DBS) and whole blood. An analytical evaluation using an HIV-1 secondary standard yielded limits of detection of 514, 710, and 1,090 HIV RNA copies/ml for EDTA plasma, whole blood, and DBS, respectively. The precision and reproducibility of HIV-1 detection was equivalent for DBS and whole blood. Inclusivity was demons...

  2. The two envelope membrane glycoproteins of Tomato spotted wilt virus show differences in lectin-binding properties and sensitivities to glycosidases

    International Nuclear Information System (INIS)

    Tomato spotted wilt virus (TSWV, Genus: Tospovirus, Family: Bunyaviridae) is a major constraint to the production of several different crops of agronomic and horticultural importance worldwide. The amino acid sequence of the two envelope membrane glycoproteins, designated as GN (N-terminal) and GC (C-terminal), of TSWV contain several tripeptide sequences, Asn-Xaa-Ser/Thr, suggesting that the proteins are N-glycosylated. In this study, the lectin-binding properties of the viral glycoproteins and their sensitivities to glycosidases were examined to obtain information on the nature of potential oligosaccharide moieties present on GN and GC. The viral proteins were separated by sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) and probed by affinoblotting using a battery of biotinylated lectins with specificity to different oligosaccharide structures. GC showed strong binding with five mannose-binding lectins, four N-acetyllactosamine-binding lectins and one fucose-binding lectin. GN was resolved into two molecular masses and only the slow migrating form showed binding, albeit to a lesser extent than GC, with three of the five mannose-binding lectins. The N-acetyllactosamine- and fucose-specific lectins did not bind to either molecular mass form of GN. None of the galactose-, N-acetylgalactosamine-, or sialic acid-binding lectins tested showed binding specificity tectins tested showed binding specificity to GC or GN. Treatment of the denatured virions with endoglycosidase H and peptide:N-glycosidase F (PNGase F) resulted in a significant decrease in the binding of GC to high mannose- and N-acetyllactosamine-specific lectins. However, no such differences in lectin binding were apparent with GN. These results indicate the presence of N-linked oligosaccharides of high mannose- and complex-type on GC and possibly high mannose-type on GN. Differences in the extent of binding of the two envelope glycoproteins to different lectins suggest that GC is likely to be more heavily N-glycosylated than GN. No evidence was observed for the presence of O-linked oligosaccharides on GN or GC

  3. An Invertebrate Warburg Effect: A Shrimp Virus Achieves Successful Replication by Altering the Host Metabolome via the PI3K-Akt-mTOR Pathway

    OpenAIRE

    Su, Mei-An; Huang, Yun-Tzu; Chen, I-Tung; Lee, Der-Yen; Hsieh, Yun-Chieh; Li, Chun-Yuan; Ng, Tze Hann; Liang, Suh-Yuen; LIN, SHU-YU; Huang, Shiao-Wei; Chiang, Yi-An; Yu, Hon-Tsen; Khoo, Kay-Hooi; Chang, Geen-Dong; Lo, Chu-Fang

    2014-01-01

    In this study, we used a systems biology approach to investigate changes in the proteome and metabolome of shrimp hemocytes infected by the invertebrate virus WSSV (white spot syndrome virus) at the viral genome replication stage (12 hpi) and the late stage (24 hpi). At 12 hpi, but not at 24 hpi, there was significant up-regulation of the markers of several metabolic pathways associated with the vertebrate Warburg effect (or aerobic glycolysis), including glycolysis, the pentose phosphate pat...

  4. WSSV ie1 promoter is more efficient than CMV promoter to express H5 hemagglutinin from influenza virus in baculovirus as a chicken vaccine

    OpenAIRE

    Yu Li; Ho YuenFern; He Fang; Kwang Jimmy

    2008-01-01

    Abstract Background The worldwide outbreak of influenza A (H5N1) viruses among poultry species and humans highlighted the need to develop efficacious and safe vaccines based on efficient and scaleable production. Results White spot syndrome virus (WSSV) immediate-early promoter one (ie1) was shown to be a stronger promoter for gene expression in insect cells compared with Cytomegalovirus immediate-early (CMV) promoter in luciferase assays. In an attempt to improve expression efficiency, a rec...

  5. Import risk analysis: A case study of white shrimp in Thailand

    Directory of Open Access Journals (Sweden)

    Supamattaya, K.

    2005-02-01

    Full Text Available As the culture industry of black tiger shrimp in Thailand has encountered several problems causing unsuccessful shrimp culture over decades, a new non-indigenous marine species, i.e. Pacific white shrimp (Penaeus vannamei was imported into Thailand as an alternation. However, imported white shrimp may carry some infectious agents which pose serious threats on aquatic species native to Thailand. Therefore, in the present study the import risk analysis (IRA was conducted to identify any hazard and estimate the risk presented by importation of white shrimp. The process involves the risk analysis steps of hazard identification and characterization, risk assessment and risk management. The risks associated with individual diseases and disease agents of white shrimp have been evaluated. Risk assessment conducted using risk evaluation matrix indicated high risk of Taura syndrome virus (TSV, White spot syndrome virus (WSSV and Infectious hypodermal and haematopoietic necrosis virus (IHHNV in imported white shrimp. The iterative process of risk management leads to a set of acceptable measures or strategies for each identified hazard for which the unrestricted risk is considered higher than appropriate level of protection. These measures or strategies will reduce risk to a level that is considered acceptable. Where measures or strategies that reduce the risk associated with a particular hazard to an acceptable level cannot be identified, permission to import the relevant commodity will be denied. The measures implemented in the control of white shrimp imports constitute quarantine and health certificate issued by exporting countries.

  6. SPOT Program

    Science.gov (United States)

    Smith, Jason T.; Welsh, Sam J.; Farinetti, Antonio L.; Wegner, Tim; Blakeslee, James; Deboeck, Toni F.; Dyer, Daniel; Corley, Bryan M.; Ollivierre, Jarmaine; Kramer, Leonard; Zimmerman, Patrick L.; Khatri, Reshma

    2010-01-01

    A Spacecraft Position Optimal Tracking (SPOT) program was developed to process Global Positioning System (GPS) data, sent via telemetry from a spacecraft, to generate accurate navigation estimates of the vehicle position and velocity (state vector) using a Kalman filter. This program uses the GPS onboard receiver measurements to sequentially calculate the vehicle state vectors and provide this information to ground flight controllers. It is the first real-time ground-based shuttle navigation application using onboard sensors. The program is compact, portable, self-contained, and can run on a variety of UNIX or Linux computers. The program has a modular objec-toriented design that supports application-specific plugins such as data corruption remediation pre-processing and remote graphics display. The Kalman filter is extensible to additional sensor types or force models. The Kalman filter design is also strong against data dropouts because it uses physical models from state and covariance propagation in the absence of data. The design of this program separates the functionalities of SPOT into six different executable processes. This allows for the individual processes to be connected in an a la carte manner, making the feature set and executable complexity of SPOT adaptable to the needs of the user. Also, these processes need not be executed on the same workstation. This allows for communications between SPOT processes executing on the same Local Area Network (LAN). Thus, SPOT can be executed in a distributed sense with the capability for a team of flight controllers to efficiently share the same trajectory information currently being computed by the program. SPOT is used in the Mission Control Center (MCC) for Space Shuttle Program (SSP) and International Space Station Program (ISSP) operations, and can also be used as a post -flight analysis tool. It is primarily used for situational awareness, and for contingency situations.

  7. Southern rice black-streaked dwarf virus: a white-backed planthopper-transmitted fijivirus threatening rice production in Asia

    OpenAIRE

    Zhou, Guohui; Xu, Donglin; Xu, Dagao; Zhang, Maoxin

    2013-01-01

    Southern rice black-streaked dwarf virus (SRBSDV), a non-enveloped icosahedral virus with a genome of 10 double-stranded RNA segments, is a novel species in the genus Fijivirus (family Reoviridae) first recognized in 2008. Rice plants infected with this virus exhibit symptoms similar to those caused by Rice black-streaked dwarf virus. Since 2009, the virus has rapidly spread and caused serious rice losses in East and Southeast Asia. Significant progress has been made in recent years in unders...

  8. A comparison of ectoparasite prevalence and occurrence of viral haemorrhagic septicemia virus (VHSV) in whiting Merlangius merlangus euxinus / Una comparación de la prevalencia de ectoparásitos y ocurrencia de virus septicémico hemorrágico viral (VHSV) en el merlán Merlangius merlangus euxinus

    Scientific Electronic Library Online (English)

    Hamdi, Ogut; Nejmettin, Cavus.

    2014-04-01

    Full Text Available Se examinaron 784 individuos del merlán Merlangius merlangus euxinus para su análisis virológico y parasitológico para establecer el efecto de la intensidad media de ectoparásitos sobre la ocurrencia del virus septicémico hemorrágico viral (VHSV) en peces del Mar Negro. El homogenizado de órganos in [...] ternos (riñón, bazo e hígado) fue inoculado con células BF-2. Las células que mostraron efecto citopático (CPE) fueron probadas con ELISA para determinar la especie de virus. Las intensidades y prevalencia promedio de los ectoparásitos (Trichodina spp. y Gyrodactylus alviga) fueron determinadas sobre las branquias de los mismos peces. Los peces infectados con VHSV presentaron intensidades similares de Trichodina spp. (10,0 ± 1,8) a aquellos libres de VHSV (7,2 ± 1,0). Las intensidades medias de Gyrodactylus alviga fueron similares en los peces infectados con VHSV (1,8 ± 0,1) que en aquellos no infectados (3,3 ± 0,2). Estos resultados indican que los ectoparásitos no son los únicos responsables de la ocurrencia observada de VHSV. Otros factores, como el estrés del desove, fluctuaciones imprevistas en los parámetros de la calidad del agua por sí solo, o aditivamente con los ectoparásitos, pueden ser responsables de la ocurrencia de VHSV en el merlán. Abstract in english Seven hundred and eighty-four whiting (Merlangius merlangus euxinus) were individually sampled for virological and parasitological examination to assess the effect of ectoparasite mean intensity on the occurrence of viral haemorrhagic septicemia virus (VHSV) in whiting in the Black Sea. The pooled h [...] omogenates of internal organs (kidney, spleen and liver) were inoculated onto BF_2 cells. The cells exhibiting cytopathic effect (CPE) were tested with ELISA to determine the virus species. The mean intensities and prevalence of ectoparasites (Trichodina spp. and Gyrodactylus alviga) were also determined on the gills of the same fish. The whiting infected with VHSV contained similar counts of Trichodina spp. (10.0 ± 1.8) than VHSV-free whiting (7.2 ± 1.0). The mean intensities of Gyrodactylus alviga were similar on the VHSV infected (1.8 ± 0.1) than uninfected fish (3.3 ± 0.2). The results indicate that ectoparasites were not alone responsible for the observed occurrence of VHSV. Other factors, such as spawning stress, sudden fluctuations in water quality parameters alone or additively to ectoparasites may be responsible for the occurrence of VHSV in whiting.

  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 por ácaros Brevipalpus (VTB. Brevipalpus phoenicis coletados de C. x speciosum sintomático e transferidos para plantas sadias de C. x speciosum reproduziram as lesões. O ácaro também transmitiu o patógeno para C. thomsonae, Gomphrena globosa, Hibiscus cannabinus, H. coccineus, H. schizopetalus, Salvia leucantha, Spathiphyllum wallasi e Tetragonia expansa, as quais exibiram manchas cloróticas e/ou necróticas. O vírus também foi transmitido mecanicamente para Chenopodium amaranticolor, C. quinoa, G. globosa, H. cannabinus, H. coccineus e T. expansa, além de C. x speciosum. Plantas de C. amaranticolor e C. quinoa mantidas a 28 - 30ºC desenvolveram infecção sistêmica. Em todos os tecidos sintomáticos das plantas-teste inoculadas, examinados ao microscópio eletrônico, foram encontrados efeitos citopáticos do tipo nuclear causado por VTB. O vírus foi purificado a partir de folhas com infecção sistêmica de C. amaranticolor e C. quinoa. Injeções de preparações purificadas em coelho geraram um anti-soro policlonal que reagiu especificamente com o antígeno homólogo em teste de ELISA. As evidências obtidas indicam que as manchas cloróticas do Clerodendrum estão associadas a um VTB do tipo nuclear, tentativamente denominado de vírus da mancha clorótica do Clerodendrum (Clerodendrum chlorotic spot virus- ClCSV.

  10. Spot Thirteen

    Science.gov (United States)

    This interactive applet affords students the opportunity to work systematically in making and testing hypotheses. Learners receive points by choosing 13 spots on a 5x5 grid. They must figure out the scoring system in order to maximize their scores. Ideas for implementation, extension and support are included along with a printable sheet of grids.

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

    OpenAIRE

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

    2012-01-01

    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 virulentus spores in a nutrient film technique (NFT) system. Lettuce plants showing BVD are generally infected by two viruses: Mirafiori lettuce big-vein virus (MiLBVV, genus Ophiovirus) and LBVaV....

  12. Analysis of the populations genetic variability of Tagosodes orizicolus (Homoptera: Delphacidae), virus vector of the rice white leaf

    International Nuclear Information System (INIS)

    Tagosodes orizicolus (Homoptera: Delphacidae), is a monophagous insect of the rice and virus vector of the white leaf (RHBV). It is distributed in America Central, El Caribe, part of the America del Sur and in Costa Rica it is in all the producing zones of rice. The genetic variability was analyzed by means of RAPD-PCR of individuals from three populations of Costa Rica: Parrita, Guanacaste (Liberia) y San Carlos (Santa Clara), that they are found separated geographically. The technique consisted of amplifying regions at random of the genome of these insects utilizing five primers. A total of 72 polymorphic bands were obtained, that upon being analyzed statistically by means of the multivariate analysis program of numerical taxonomy could show a clear genetic distancing among said populations. The genetic distance observed in the molecular analysis can be explained for the climatic and/or geographical isolation of the populations or by the incident of Wolbachia, riquettsia that induces cytoplasmic sterility in insects. These symbionts are transmitted of generation in generation, of the mother to their offspring and they cause reproductive alterations as cytoplasmic incompatibility, parthenogenesis and feminization. The presence of Wolbachia was determined by means of transmission electronic microscopy being observed in the greasy and muscular weave of the abdomen of T. orizicolus. Rickettsias present pleomorphic morphology and form small groups, that are characterized form small groups, that are characterized for the presence of electrondense material semidetached to the cell wall with an interior electronlucent. Its size in transverse cuts ranged between 520 nm of length X 470 nm of width. In addition, its presence was detected by means of the amplification by PCR of the genomic DNA of the insects; a specific primer for the DNA ribosomal 16S of Wolbachia was utilized for it. A 86% of insects of the San Carlos population were positive, a 96% was determined for Guanacaste, a 37% for Parrita and a 100% for Colombia. Individual insects originating from populations of T. orizicolus of the zones of Parrita and Guanacaste, maintained in greenhouse, they were crossed among them in the greenhouse. Of the crossing infected males with uninfected females progeny was not obtained, what it seems to indicate that is cytoplasmic incompatibility unidirectional. Also, a reduction or a drastic absence of offspring was observed in those crossings where both insects were infected, what could be explained by the incidence of more than a race of the bacterium (bidirectional mutual incompatibility). This last phenomenon could be a head in part of the genetic isolation observed among populations of T. orizicolus originating from different localities. (author)

  13. Symantec: Virus Alerts and Hoaxes

    Science.gov (United States)

    This website by Symantec (makers of Norton AntiVirus) provides information on the latest virus threats, security advisories, updates for Symantec products and removal tools, as well as some basic information on viruses. The Reference Area includes FAQ, a Glossary, Newsletter, White Papers, a section where you can Submit Virus Samples, postings of Hoaxes, a Security Database, Virus Encyclopedia, and Virus Calendar.

  14. 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)

    Scientific Electronic Library Online (English)

    Elliot Watanabe, Kitajima; Karen Sumire, Kubo; Paulo de Tarso Oliveira, Ferreira; Berenice Kussumoto de, Alcântara; Alessandra Jesus, Boari; Renata Takassugi, Gomes; Juliana, Freitas-Astua; Jorge Alberto Marques, Rezende; Gilberto José de, Morais; Renato Barbosa, Salaroli.

    2008-02-01

    Full Text Available 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 por ácaros Brevipalpus (VTB). Brevipalpus phoenicis coletados de C. x speciosum sintomático e transferidos para plantas sadias de C. x speciosum reproduziram as lesões. O ácaro também transmitiu o patógeno para C. thomsonae, Gomphrena globosa, Hibiscus cannabinus, H. coccineus, H. schizopetalus, Salvia leucantha, Spathiphyllum wallasi e Tetragonia expansa, as quais exibiram manchas cloróticas e/ou necróticas. O vírus também foi transmitido mecanicamente para Chenopodium amaranticolor, C. quinoa, G. globosa, H. cannabinus, H. coccineus e T. expansa, além de C. x speciosum. Plantas de C. amaranticolor e C. quinoa mantidas a 28 - 30ºC desenvolveram infecção sistêmica. Em todos os tecidos sintomáticos das plantas-teste inoculadas, examinados ao microscópio eletrônico, foram encontrados efeitos citopáticos do tipo nuclear causado por VTB. O vírus foi purificado a partir de folhas com infecção sistêmica de C. amaranticolor e C. quinoa. Injeções de preparações purificadas em coelho geraram um anti-soro policlonal que reagiu especificamente com o antígeno homólogo em teste de ELISA. As evidências obtidas indicam que as manchas cloróticas do Clerodendrum estão associadas a um VTB do tipo nuclear, tentativamente denominado de vírus da mancha clorótica do Clerodendrum (Clerodendrum chlorotic spot virus- ClCSV). Abstract in english 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 vir [...] uses (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).

  15. Barreras de maíz en una estrategia de manejo integral para controlar epidemias del virus mancha anular del papayo (PRSV-P) / Corn barriers in an integrated management strategy to control epidemics of papaya ring spot virus (PRSV-P)

    Scientific Electronic Library Online (English)

    Elías, Hernández-Castro; J. Antonio, Villanueva-Jiménez; J. Antonio, Mora-Aguilera; Cristian, Nava-Díaz.

    2010-05-01

    Full Text Available Las epidemias ocasionadas por el virus de la mancha anular del papayo (PRSV-P) (Carica papaya L.) se han logrado retrasar mediante un sistema de manejo integral (MIP), aunque no se ha medido la efectividad individual de algunos componentes. Para ello se evaluó el establecimiento de barreras de maíz [...] (Zea mays L.) en el control del PRSV-P en Veracruz, México. El MIP incluyó la protección del vivero con malla de polipropileno, 2743 plantas ha-1, la erradicación de plantas enfermas, la fertilización mineral y el control de arvenses. Los tratamientos fueron: manejo integral con barreras de maíz (MIP), manejo integral sin barreras (MIZ) y manejo regional (MR, 1600 plantas ha-1, con insecticidas, sin barreras ni eliminación de plantas enfermas). El modelo logístico explicó adecuadamente la incidencia en MIP, MIZ y MIR, y la severidad en MIZ (R²=0.92 a 0.97); el exponencial y Gompertz explicaron la severidad (R²=0.92 y 0.98) en el MIP y MR. La mayor tasa epidémica se observó en el MR (0.054), mientras que el MIP la redujo en 42 % (0.031). Las barreras indujeron menor incidencia y severidad promedio (Yp-MIP= 18.4 y 7.1 %; Yp-MIZ= 23.4 y 11.7 %) y fnal (Yf-MIP= 81.0 y 35.7 %; Yf-MIZ= 94.11 y 43.0 %). La severidad máxima a 238 d después del trasplante (ddt) se observó en MR (69.0 %) y la menor en MIP (35.7 %). La ausencia de barreras incrementó las epidemias; en MR la máxima incidencia (100 %) y severidad (60 %) ocurrió 175 ddt, mientras que en MIP 238 ddt (incidencia 80 %, severidad 35 %). Las barreras mejoraron la sanidad del MIP en 14.0 % (Yf-MIP = 81.0 % Abstract in english Te epidemics caused by papaya ring spot virus (PRSV-P) (Carica papaya L.) have been delayed by an integral management system (MIP), although the individual efectiveness of some components has not been measured. Te establishment of corn (Zea mays L.) barriers was evaluated in the control of PRSV-P in [...] Veracruz, Mexico. Te MIP included the protection of the nursery with polypropylene mesh, 2743 plants ha-1, eradication of diseased plants, mineral fertilization and weed control. Treatments were: integral management with corn barriers (MIP), integral management without barriers (MIZ) and regional management (MR, 1600 plants ha-1, using insecticides, without barriers and elimination of diseased plants). Te logistic model adequately explained the incidence in MIP, MIZ and MR, and severity (R²=0.92 to 0.97) in MIZ; the exponential and Gompertz models explained severity (R²= 0.92 and 0.98) in MIP and MR. Te highest epidemic rate was observed in MR (0.054), while MIP reduced it by 42 % (0.031). Barriers induced lower incidence and mean severity (Yp -MIP = 18.4 and 7.1 %; Yp-MIZ = 23.4 and 11.7 %) and final (Yf-MIP = 81.0 and 35.7 %; Yf-MIZ = 94.11 and 43.0 %). Maximum severity at 238 d after transplant (dat) was observed in MR (69.0 %) and the lowest in MIP (35.7 %). Te absence of barriers increased the epidemics; in MR the maximum incidence (100 %) and severity (60 %) occurred 175 dat, while in MIP, 238 dat (incidence 80 %, severity 35 %). Barriers improved papaya health under MIP by 14 % (Yf-MIP =81.0 %

  16. Presence and Distribution of Tobacco Viruses in Montenegro

    OpenAIRE

    Jelena Zindovi?; Nataša Duki?; Aleksandra Bulaji?; Jelena Latinovi?; Ivana ?eki?; Bojan Duduk; Branka Krsti?

    2007-01-01

    Seven important tobacco viruses were investigated in Montenegro in 2005: Tobacco Mosaic Virus (TMV), Tomato Spotted Wilt Virus (TSWV), Cucumber Mosaic Virus (CMV), Potato Virus Y (PVY), Alfalfa Mosaic Virus (AMV), Tobacco Ring Spot Virus (TRSV) and Potato Virus X(PVX). This investigation included sample collection from four tobacco growing regions in Montenegro and their serological testing by DAS-ELISA test. Presence of different strains of PVY was investigated as well using DAS ELISA test w...

  17. Presence and Distribution of Tobacco Viruses in Serbia

    OpenAIRE

    Nataša Duduk; Aleksandra Bulaji?; Janoš Berenji; Ivana ?eki?; Bojan Duduk; Branka Krsti?

    2006-01-01

    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 invest...

  18. Penetración de tres adhesivos en lesiones interproximales de caries de mancha blanca: estudio in vitro / Penetration of three adhesives in proximal white- spot carious lesions: an in vitro study

    Scientific Electronic Library Online (English)

    Olga Lucía, Zarta; Alba Lucía, Zuluaga Valencia; María Cristina, Huertas Díaz; Juan Sebastián, Lara; Ingrid Katherine, Quintero; Marisol, Téllez; Margarita, Úsuga; Stefanía, Martignon.

    2012-12-01

    Full Text Available INTRODUCCIÓN: el tratamiento de lesiones interproximales de caries temprana representa un reto en la concepción moderna de la cariología y la operatoria. Como respuesta, además del diagnóstico temprano, emergen nuevos materiales para controlar la progresión de la caries temprana. El objetivo de este [...] estudio fue evaluar in vitro el grado de penetración de tres adhesivos: Excite (Ivoclar-Vivadent), Prime & Bond NT (Dentsply) y Single Bond (3M-ESPE), en el sellado de lesiones interproximales no cavitacionales de mancha blanca. MÉTODOS: 117 premolares humanos extraídos con lesión interproximal de mancha blanca Icdas-II 2 montados en yeso, simulando la situación en cavidad oral, fueron asignados aleatoriamente a uno de los tres grupos; se sellaron las lesiones con cada material previamente mezclado con azul de metileno; se obtuvieron secciones longitudinales de 250 µm y se analizaron en fotografías obtenidas bajo estereomicroscopía. Se valoró cubrimiento total de la lesión, profundidad histológica de la lesión en zonas y en micrómetros y, penetración máxima del adhesivo. RESULTADOS: se analizaron 116 dientes. Se encontró cubrimiento total de la lesión en 52,5%; 75% se ubicaron en la mitad interna del esmalte, con profundidad promedio de 854 ± 493 µm. La máxima penetración promedio del adhesivo en la lesión fue de 697 ± 412 µm. No hubo diferencias estadísticamente significativas entre grupos para las variables (Kruskal Wallis y correlación de Spearman; p > 0,05). CONCLUSIONES: estos tres adhesivos de baja viscosidad, con alto contenido de TEGDMA y HEMA, lograron el cubrimiento y la penetración de material para el sellado de lesiones tempranas de caries interproximal. Abstract in english INTRODUCTION: the treatment of early proximal carious lesions represents a challenge to modern conceptions of cariology and operative dentistry. The possible solutions include, besides an early diagnosis, development of new materials to control early caries progression. The objective of this in vitr [...] o study was to evaluate the degree of penetration of three adhesives: Excite (Ivoclar-Vivadent), Prime & Bond NT (Dentsply), and Single Bond (3M-ESPE), in the process of sealing white-spot non-cavitated proximal lesions. METHODS: 117 extracted human premolars with ICDAS 2 white-spot non-cavitated proximal lesions mounted on plaster, to simulate oral cavity conditions, were randomly sorted out in each of the three groups. The lesions were sealed with each of the materials previously mixed with methylene blue; longitudinal sections of 250 µm were obtained, and they were later analyzed on photographs taken with a stereo microscope. These variables were evaluated: total lesion coating, lesion's histologic depth (in micrometers), and the adhesive's maximum penetration. RESULTS: a total of 116 teeth were analyzed. Entire lesion coating was found in 52.5% of the cases; 75% were located in the enamel's internal half, with an average depth of 854 ± 493 µm. The adhesive's maximum average lesion penetration was 697 ± 412 µm. No statistical significant differences were found among the groups (Kruskal Wallis and Spearman correlation; p > 0.05). CONCLUSION:these three low-viscosity adhesives with high amounts of TEGDMA and HEMA effectively achieved covering and penetrating of the material for sealing early proximal carious lesions.

  19. A possible dominant white gene in Jersey cattle

    Directory of Open Access Journals (Sweden)

    Sponenberg D Phillip

    2001-01-01

    Full Text Available Abstract A white heifer ("Snow" was born in 1991 from coloured registered Jersey parents. She produced six calves sired by coloured Jersey bulls: three white bull calves, two white heifer calves, and one coloured bull calf. One of the white bull calves was mated with 40 Hereford × Friesian yearling heifers (white face, predominantly black body with some white patches. The 38 resulting calves included 16 white and 22 coloured calves. Twelve of the 16 white calves were heifers and four were bulls. Red or black spotting was recorded on some white calves. The results are consistent with an autosomal dominant mutant causing the white phenotype. The mutation appears to have arisen spontaneously in Snow, then passing to her white progeny and white grand-progeny. The white individuals varied from entirely white in a few cases, to most having some residual small areas of red or black pigmentation in patterns not typical of other reported white spotting patterns of cattle.

  20. Activating Transcription Factor 4 and X Box Binding Protein 1 of Litopenaeus vannamei Transcriptional Regulated White Spot Syndrome Virus Genes Wsv023 and Wsv083

    OpenAIRE

    Li, Xiao-Yun; Pang, Li-Ran; Chen, Yong-gui; Weng, Shao-ping; Yue, Hai-Tao; Zhang, Ze-Zhi; Chen, Yi-Hong; He, Jian-guo

    2013-01-01

    In response to endoplasmic reticulum (ER) stress, the signaling pathway termed unfolded protein response (UPR) is activated. To investigate the role of UPR in Litopenaeus vannamei immunity, the activating transcription factor 4 (designated as LvATF4) which belonged to a branch of the UPR, the [protein kinase RNA (PKR)-like ER kinase, (PERK)]-[eukaryotic initiation factor 2 subunit alpha (eIF2?)] pathway, was identified and characterized. The full-length cDNA of LvATF4 was 1972 bp long, with a...

  1. KARAKTERISASI DAN KLONING GEN PENGKODE VP28 WHITE SPOT SYNDROME VIRUS ( WSSV) ISOLAT INDONESIA SEBAGAI KANDIDAT VAKSI REKOMBINAN UNTUK PENGENDALIAN PENYAKIT BINTIK PUTIH PADA UDANG WINDU (PENAEUS MONODON )

    OpenAIRE

    Sriwulan; Irmawati; Debbi S.Retnoningrum; Ernawati G.R; Hilal Anshary

    2007-01-01

    Penelitian bertujuan untuk mendapatkan gen VP28 WSSV dari udang yang terinfeksi WSSV, Karakterisasi dan kloning gen VP 28. Sampel yang digunakan adalah udang yang terinfeksi WSSV pada seluruh sentra produksi udang di Sulawesi Selatan dan ekstrak DNA udang yang positif WSSV dengan IQ 2000. Untuk mendapatkan gen VP28 dilakukan dilakukan ektraksi DNA dengan metode Wizard, karakterisasi dilakukan dengan sekuensing dan kloning dengan menggunakan vektor kloning p-GEMT yang dilakukan untuk memperban...

  2. Prevalence of Taura syndrome virus (TSV and Infectious hypodermal and haematopoietic necrosis virus (IHHNV in white shrimp (Penaeus vannamei populations and susceptibility to infection of some aquatic species native to Thailand

    Directory of Open Access Journals (Sweden)

    Supamattaya, K.

    2005-02-01

    Full Text Available This study aimed to survey the prevalence of some infectious diseases e.g. Taura syndrome virus (TSV and Infectious hypodermal and haematopoietic necrosis virus (IHHNV in white shrimp (Penaeus vannamei populations and to assess the impact of such infectious agents to indigenous aquatic animals in Thailand. Samples of both larval and juvenile or adult shrimp from each region of the country were collected and screened for TSV and IHHNV using the polymerase chain reaction (PCR technique. Viruses isolated from affected shrimp were used for determine the susceptibility to infection of some aquatic species native to Thailand.A total of 163 samples of larval shrimp from hatcheries were screened. The results showed infection with TSV and IHHNV in 3.68 and 44.17%, respectively. As high as 7.32% TSV infection was detected in shrimp samples collected from the South Eastern coast, followed by the Eastern and Central regions with percentages of 5.56 and 4.53, respectively. Shrimp with the highest rate of IHHNV infection, 55.56% were collected from the Eastern region. A total of 192 samples of shrimp reared in grow-out ponds were also collected. The results showed shrimp were infected with TSV and IHHNV with percentages of 6.67 and 67.19, respectively. The highest prevalence of IHHNV (up to 90% was found in samples collected from the lower Southern region. The highest prevalence of TSV infection (11.29% was reported in shrimp from the Central region. A study of the susceptibility to TSV and IHHNV infection of some indigenous aquatic species of Thailand was also carried out. The results showed many aquatic species native to Thailand e.g. black tiger shrimp (Penaeus monodon, speckled shrimp (Metapenaeus monoceros, dwarf prawn (Macrobrachium equideus, krill (Acetes sp., mantis lobster (Chloridopsis immaculatus, freshwater prawn (Macrobrachium lanchesteri and M. rosenbergii, mangrove crab (Sesarma sp. and mud crab (Scylla serrata were susceptible to viruses and died due to infection. The mortality of affected species associated with a causative agent was confirmed in most species, except the mud crab and freshwater prawn (Marcrobrachium rosenbergii. However, viral particles can be still detected in surviving animals 10 days after infection. The results of this study will be a helpful tool employed in establishing measures on disease control and reduction of risk with the importation of white shrimp broodstock.

  3. Viral MicroRNAs Targeting Virus Genes Promote Virus Infection in Shrimp In Vivo

    Science.gov (United States)

    He, Yaodong; Yang, Kai

    2014-01-01

    Viral microRNAs (miRNAs), most of which are characterized in cell lines, have been found to play important roles in the virus life cycle to avoid attack by the host immune system or to keep virus in the latency state. Viral miRNAs targeting virus genes can inhibit virus infection. In this study, in vivo findings in Marsupenaeus japonicus shrimp revealed that the viral miRNAs could target virus genes and further promote the virus infection. The results showed that white spot syndrome virus (WSSV)-encoded miRNAs WSSV-miR-66 and WSSV-miR-68 were transcribed at the early stage of WSSV infection. When the expression of WSSV-miR-66 and WSSV-miR-68 was silenced with sequence-specific anti-miRNA oligonucleotides (AMOs), the number of copies of WSSV and the WSSV-infected shrimp mortality were significantly decreased, indicating that the two viral miRNAs had a great effect on virus infection. It was revealed that the WSSV wsv094 and wsv177 genes were the targets of WSSV-miR-66 and that the wsv248 and wsv309 genes were the targets of WSSV-miR-68. The data demonstrate that the four target genes play negative roles in the WSSV infection. The targeting of the four virus genes by WSSV-miR-66 and WSSV-miR-68 led to the promotion of virus infection. Therefore, our in vivo findings show a novel aspect of viral miRNAs in virus-host interactions. PMID:24198431

  4. Café-au-lait spots in schoolchildren.

    OpenAIRE

    Burwell, R G; James, N J; Johnston, D I

    1982-01-01

    This paper reports a study of café-au-lait spots of a minimum diameter of 1 cm in 732 white schoolchildren. Three groups were identified, according to the number of café-au-lait spots on each child: (1) those with none (74%), (2) those with fewer than 5 (25%), and (3) those with at least 5 (5 children, 2 considered to be normal, and 3 siblings each presumed to have neurofibromatosis, one having died from leukaemia). Excluding the last group, the number of café-au-lait spots in the sample was ...

  5. 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

    Full Text Available Entre cêrca de 200 variedades de feijoeiro ensaiadas, a Manteiga foi mais sensitiva aos vírus de vira-cabeça e da necrose branca, formando, nas fôlhas primárias, lesões locais adequadas para contagens. Com o vírus de vira-cabeça as lesões locais apareceram de 3 -6 dias após as inoculações, sob a forma de manchas cloróticas, às vezes com anéis necróticos; com o vírus da necrose branca as lesões se manifestaram em 1 - 4 dias, sob a forma de pontuações necróticas, pequenos anéis necróticos ou, em casos de certas estirpes do vírus, como pintas cloróticas. Feijoeiros com as fôlhas primárias tendo 2/3 de sua expansão ou mais desenvolvidas deram melhores resultados para o vírus de vira-cabeça; para o vírus da necrose branca os melhores resultados foram obtidos com fôlhas que tinham 2/3 ou ligeiramente menos de sua expansão total. O vírus de vira-cabeça não se tornou sistêmico em plantas inoculadas mecanicamente. A maioria das estirpes do vírus da necrose branca também não se tornou sistêmica em feijoeiros inoculados mecanicamente. Uma estirpe amarela deste tem a capacidade de se tornar sistêmica em feijoeiros sob determinadas condições. O sulfito de sódio em solução 0,0125 M, usado na extração do inóculo, aumentou ligeiramente o número de lesões formadas nas fôlhas do feijoeiro Manteiga pelo vírus de vira-cabeça; aumento consideravelmente maior foi obtido com a extração em presença de búfer de fosfato 0,1 M com pH 7, sendo êsse efeito atribuído a um aumento da sensitividade da planta-teste. A mistura de ambos causou aumento igual ou ligeiramente menor que o búfer só. Para o vírus da necrose branca a presença de sulfito de sódio 0,0125 M durante a extração aumentou consideravelmente o número de lesões, sendo esse efeito atribuído à diminuição na perda de atividade do vírus, devida à oxidação; o búfer sozinho ocasionou pequeno aumento em comparação com a testemunha, mas a 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 viruses. Bean plants kept for 48 hours under reduced light gave a higher number of lesions th

  6. Scientific Opinion on the risk to plant health posed by Tomato spotted wilt virus to the EU territory with identification and evaluation of risk reduction options

    Directory of Open Access Journals (Sweden)

    EFSA Panel on Plant Health (PLH

    2012-12-01

    Full Text Available The Panel on Plant Health conducted a pest risk assessment for Tomato spotted wilt virus (TSWV including the identification and evaluation of risk reduction options. The plants for planting pathway is considered the most important one and rated as likely, with medium uncertainty. TSWV, reported in many Member States, has one of the widest host range for plant viruses and has three widely distributed thrips vector species in the risk assessment area. For countries falling under the provisions of 2000/29/EC Annex IIAII legislation, the probability of establishment is rated as very likely, with low uncertainty. For Finland and Sweden, protected zone under the 2000/29/EC Annex IB legislation, the probability of establishment is rated unlikely to moderately likely. Spread can occur through the activity of thrips vectors and through trade of infected hosts. The probability of spread in Member States under Annex IIAII provisions is evaluated as likely, with low uncertainty. In Finland and Sweden, spread is rated as unlikely to moderately likely, with medium uncertainty. Since TSWV may cause both yield and quality losses in a wide range of hosts, a major impact is predictable in the absence of appropriate control measures, with medium uncertainty. The risk reduction options evaluated by the Panel are those addressing either the sanitary status of the propagation material (including the provisions of 2000/29/EC Annex IIAII and Annex IB, statutory certification system and the use of voluntary industry standards or those addressing the control of TSWV epidemic development through actions directed at the virus or at its vectors (including cultural methods to reduce crop infestation and chemical or biological control of thrips vectors. Limitations of the current measures and the potential consequences of repealing the current legislation are discussed.

  7. A multiplex nested PCR assay for simultaneous detection of Corchorus golden mosaic virus and a phytoplasma in white jute (Corchorus capsularis L.).

    Science.gov (United States)

    Biswas, C; Dey, P; Satpathy, S

    2013-05-01

    A multiplex nested PCR assay was developed by optimizing reaction components and reaction cycling parameters for simultaneous detection of Corchorus golden mosaic virus (CoGMV) and a phytoplasma (Group 16Sr V-C) causing little leaf and bunchy top in white jute (Corchorus capsularis). Three sets of specific primers viz. a CoGMV specific (DNA-A region) primer, a 16S rDNA universal primer pair P1/P7 and nested primer pair R16F2n/R2 for phytoplasmas were used. The concentrations of the PCR components such as primers, MgCl2 , Taq DNA polymerase, dNTPs and PCR conditions including annealing temperature and amplification cycles were examined and optimized. Expected fragments of 1 kb (CoGMV), 674 bp (phytoplasma) and 370 bp (nested R16F2n/R2) were successfully amplified by this multiplex nested PCR system ensuring simultaneous, sensitive and specific detection of the phytoplasma and the virus. The multiplex nested PCR provides a sensitive, rapid and low-cost method for simultaneous detection of jute little leaf phytoplasma and CoGMV. Based on BLASTn analyses, the phytoplasma was found to belong to the Group 16Sr V-C. PMID:23413927

  8. Functional Analysis of a Crustacean MicroRNA in Host-Virus Interactions

    OpenAIRE

    Huang, Tianzhi; Zhang, Xiaobo

    2012-01-01

    Growing evidence from mammals suggests that host microRNAs (miRNAs) play important roles in the antiviral immune response. However, the roles of invertebrate miRNAs in response to virus infection remain to be investigated. Based on our previous studies, the shrimp miR-7 was found to be upregulated in response to white spot syndrome virus (WSSV) infection. In this study, the results showed that shrimp miR-7 could target the 3?-untranslated region (3?UTR) of the WSSV early gene wsv477, implying...

  9. Retinal vasculitis in a mother and her son with human T-lymphotropic virus type 1 associated myelopathy.

    OpenAIRE

    Hayasaka, S.; Takatori, Y.; Noda, S.; Setogawa, T.; Hayashi, H.

    1991-01-01

    A 53-year-old woman had difficulty in walking, raised titres to human T-lymphotropic virus type 1 (HTLV-1) in serum and cerebrospinal fluid, and yellowish white retinal lesions and vasculitis in the right eye. Her 20-year-old son also had difficulty in walking, raised titres to HTLV-1 in serum and cerebrospinal fluid, and retinal vasculitis and multiple whitish vitreoretinal spots in both eyes.

  10. Low white blood cell count and cancer

    Science.gov (United States)

    ... the white blood cells as well as the cancer. Other causes of a low white blood cell count include: Crohn's disease Infections, such as tuberculosis (TB) or certain viruses like HIV Lupus (also ...

  11. Characterization of the yeast form symbiant of Togosodes orizicolus (Hemiptera: delphacidae), vector of the rice white leaf virus

    International Nuclear Information System (INIS)

    The purpose of this investigation was to characterize the yeast forms symbionts of Togo soles Orizicolus (YLSTo), through its morphologic description and locating in situ through microscopy of light, electronic microscopy of transmission and of sweeping and immuno microscopy. Likewise, molecular tests were carried out to classify phylogenetically the symbionts, utilizing partial sequences of the ribosomal DNA 18S. This study will permit to determine, the existence or not of interactions among the YLSTo and the insect vector, in the future. The paper of that interaction in the mechanism of trans ovarial transmission of the virus and the search of alternatives for the control of the disease in the rice. (S. Grainger)

  12. Virus infections in Gentiana species

    OpenAIRE

    Verhoeven, J. Th J.; Ding, Y. M.; Lesemann, D. E.; Vlugt, R. A. A.; Roenhorst, J. W.

    2002-01-01

    Gentiana species generally are of little importance to horticulture. This may explain that little attention has been given to virus infections in these ornamentals, so far. However, during the last couple of years the number of samples for virus diagnosis increased. The main symptoms recorded, were growth reduction, necrotic lesions and decline. The virus isolated most frequently was broad bean wilt virus 2, while the related broad bean wilt virus 1 was not detected. Impatiens necrotic spot v...

  13. Effect of three innovative culture systems on water quality and whitespot syndrome virus (WSSV) viral load in WSSV-fed Penaeus monodon cultured in indoor tanks

    OpenAIRE

    Alapide-tendencia, E. V.; Bosma, R. H.; Rose Sorio, L.

    2012-01-01

    White spot syndrome virus is the most important among the shrimp diseases. It has been devastating the shrimp industry for more than 3 decades. Previous studies reported that greater percentage of yellow colonies on thiosulfate citrate bile salt sucrose agar (yellow vibrios) in the rearing water, abundant supply of natural food such as Chlorella, and the use of the greenwater technology (GW) are some ecological ways of preventing WSSV outbreak. The aim of this study was to investigate the eff...

  14. Quantitation of infectious myonecrosis virus in different tissues of naturally infected Pacific white shrimp, Litopenaeus vannamei, using real-time PCR with SYBR Green chemistry.

    Science.gov (United States)

    da Silva, Suzianny Maria Bezerra Cabral; Pinheiro, Ana Cristina de Aguiar Saldanha; Coimbra, Maria Raquel Moura

    2011-11-01

    The Pacific white shrimp, Litopenaeus vannamei, is the most important shrimp species in volume in world aquaculture. However, in recent decades, outbreaks of diseases, especially viral diseases, have led to significant economic losses, threatening the sustainability of shrimp farming worldwide. In 2004, Brazilian shrimp farming was seriously affected by a new disease caused by the Infectious myonecrosis virus (IMNV). Thus, disease control based on rapid and sensitive pathogen detection methods has become a priority. In this study, a specific quantitation method for IMNV was developed using real-time PCR with SYBR Green chemistry and viral load of the principal target tissues of chronically infected animals was quantified. The quantitative analysis revealed that mean viral load ranged from 5.08×10(8) to 1.33×10(6)copies/?g of total RNA in the hemolymph, 5.096×10(5) to 1.26×10(3)copies/?g in the pleopods, 6.85×10(8) to 3.09×10(4)copies/?g in muscle and 8.15×10(6) to 3.90×10(3)copies/?g in gills. Different viral loads of IMNV were found with greater values in the hemolymph and muscle, followed by the pleopods and gills. PMID:21854806

  15. Transferring cucumber mosaic virus-white leaf strain coat protein gene into Cucumis melo L. and evaluating transgenic plants for protection against infections

    Energy Technology Data Exchange (ETDEWEB)

    Gonsalves, C.; Xue, B.; Yepes, M.; Fuchs, M.; Ling, K.; Namba, S. (Cornell Univ., Geneva, NY (United States). Dept. of Plant Pathology)

    1994-03-01

    A single regeneration procedure using cotyledon examples effectively regenerated five commercially grown muskmelon cultivars. This regeneration scheme was used to facilitate gene transfers using either Agrobacterium tumefaciens or microprojectile bombardment methods. In both cases, the transferred genes were from the T-DNA region of the binary vector plasmid pGA482GG/cp cucumber mosaic virus-white leaf strain (CMV-WL), which contains genes that encode neomycin phosphotransferase II (NPT II), [beta]-glucuronidase (GUS), and the CMV-WL coat protein (CP). Explants treated with pGA482GG/cpCMV-WL regenerated shoots on Murashige and Skoog medium containing 4.4 [mu]m 6-benzylaminopurine (BA), kanamycin (Km) at 150 mg[center dot]liter[sup [minus]1] and carbenicillin (Cb) at 500 mg[center dot]liter[sup [minus]1]. The authors' comparison of A. tumefaciens- and microprojectile-mediated gene transfer procedures shows that both methods effectively produce nearly the same percentage of transgenic plants. R[sub 0] plants were first tested for GUS or NPT II expression, then the polymerase chain reaction (PCR) and other tests were used to verify the transfer of the NPT II, GUS, and CMV-WL CP genes.

  16. Anticuerpos contra el virus de estomatitis vesicular en Huanganas (Tayassu Pecari) en Madre de Dios, Perú / Antibodies against vesicular estomatitis virus in white-lipped peccaries (Tayassu Pecari) in Madre de Dios, Peru

    Scientific Electronic Library Online (English)

    Susan, Carruitero H; Hermelinda, Rivera G; Mercy, Ramírez V; Juan, More B; Alfonso, Zúñiga H; Mónica, Romero S.

    Full Text Available El objetivo del presente estudio fue determinar la presencia de anticuerpos neutralizantes contra los serotipos New Jersey (NJ) e Indiana subtipo 1 (IND-1) del virus Estomatitis Vesicular (VEV) en huanganas (Tayassu pecari) de vida libre de las localidades de Boca de Manu (n=30), Concesión para la C [...] onservación Los Amigos (n=10) y La Reserva Nacional Tambopata/Parque Nacional Bahuaja Sonene (n=48) en el departamento de Madre de Dios. La presencia de anticuerpos contra el VEV fue determinado mediante la prueba de neutralización viral en las 88 muestras de suero de huanganas machos y hembras adultos de apariencia normal. El 53.4% (47/88) y 18.2% (16/88) de las muestras fue positiva a anticuerpos contra los serotipos IND-1 y NJ, respectivamente, en tanto que el 29.5 y el 2.3% de las muestras tuvieron anticuerpos neutralizantes igual o mayor a 1:32 contra los serotipos IND-1 y NJ, respectivamente. No hubo asociación significativa entre las variables presencia de anticuerpos contra el VEV y lugar de procedencia de las muestras. Abstract in english The aim of the study was to determine the presence of antibodies against New Jersey (NJ) and Indiana subtype 1 (IND-1) Vesicular Stomatitis Virus (VSV) in free-living white-lipped peccaries (Tayassu pecari) in three localities of Madre de Dios, Peru. The presence of antibodies against VSV by virus n [...] eutralization test was determined in 88 serum samples of adult male and females in apparent good health condition. Results showed that 53.4% (47/88) and 18.2% (16/88) of samples were positive to antibodies against serotypes IND-1 and NJ respectively, whereas 55.3 and 12.6% of the serum samples had neutralizing antibodies titers equal or greater than 1:32 against serotype IND-1 and NJ respectively. There was a no significant association between seropositivity of VSV and source of samples.

  17. Rocky Mountain spotted fever

    Science.gov (United States)

    ... Mountain spotted fever is caused by the bacteria Rickettsia rickettsii (R. Rickettsii) , which is carried by ticks. ... Pa.: Elsevier Mosby; 2011:chap 51. Walker DH. Rickettsia rickettsii and other spotted fever group rickettsiae (rocky ...

  18. Performance of a Novel Human Immunodeficiency Virus (HIV) Type 1 Total Nucleic Acid-Based Real-Time PCR Assay Using Whole Blood and Dried Blood Spots for Diagnosis of HIV in Infants?

    Science.gov (United States)

    Stevens, Wendy; Erasmus, Linda; Moloi, Matsidisho; Taleng, Thabo; Sarang, Somaya

    2008-01-01

    The new Cobas AmpliPrep/Cobas TaqMan HIV-1 Qual test offers advanced automation for the detection of human immunodeficiency virus type 1 (HIV-1) RNA and DNA in dried blood spots (DBS) and whole blood. An analytical evaluation using an HIV-1 secondary standard yielded limits of detection of 514, 710, and 1,090 HIV RNA copies/ml for EDTA plasma, whole blood, and DBS, respectively. The precision and reproducibility of HIV-1 detection was equivalent for DBS and whole blood. Inclusivity was demonstrated for a reference panel of HIV-1 subtypes A to N. A clinical evaluation of the Cobas AmpliPrep/Cobas TaqMan HIV-1 Qual test was performed at a center for routine diagnostics in Johannesburg, South Africa, using 1,013 clinical specimens from HIV-1 exposed children. The Amplicor HIV-1 DNA test v1.5 with the MagNApure DNA isolation procedure was used as the reference method. A total of 995 valid results for whole blood with both methods yielded 691 and 303 concordant negative and positive results for the Cobas AmpliPrep/Cobas TaqMan HIV-1 Qual test, respectively. For the 800 valid DBS specimen results, 495 and 300 concordant negative and positive results were obtained, respectively. The resulting clinical specificities and sensitivities of the new test were 100% and 99.7% for whole blood and DBS, respectively. The new test was characterized by its robustness, enhanced automation, and improved sample throughput. The Cobas AmpliPrep/Cobas TaqMan HIV-1 Qual test will support early, reliable diagnosis of HIV in children in routine laboratory settings. PMID:18923017

  19. Meteorology of Jupiter's Equatorial Hot Spots and Plumes from Cassini

    CERN Document Server

    Choi, David S; Vasavada, Ashwin R; Simon-Miller, Amy A

    2013-01-01

    We present an updated analysis of Jupiter's equatorial meteorology from Cassini observations. For two months preceding the spacecraft's closest approach, the Imaging Science Subsystem (ISS) onboard regularly imaged the atmosphere. We created time-lapse movies from this period in order to analyze the dynamics of equatorial hot spots and their interactions with adjacent latitudes. Hot spots are quasi-stable, rectangular dark areas on visible-wavelength images, with defined eastern edges that sharply contrast with surrounding clouds, but diffuse western edges serving as nebulous boundaries with adjacent equatorial plumes. Hot spots exhibit significant variations in size and shape over timescales of days and weeks. Some of these changes correspond with passing vortex systems from adjacent latitudes interacting with hot spots. Strong anticyclonic gyres present to the south and southeast of the dark areas appear to circulate into hot spots. Impressive, bright white plumes occupy spaces in between hot spots. Compact...

  20. Anatomia de lesões foliares causadas pelo vírus da Mancha Clorótica do Clerodendrum, transmitido pelo ácaro Brevipalpus phoenicis em diferentes espécies / Anatomy of leaf lesions caused by Clerodendrum Chlorotic Spot Virus (ClCSV) transmitted by the mite Brevipalpus phoenicis in different host species

    Scientific Electronic Library Online (English)

    Renata Takassugui, Gomes; Elliot Watanabe, Kitajima; Francisco André Osamu, Tanaka; João Paulo Rodrigues, Marques; Beatriz, Appezzato-da-Glória.

    2010-12-01

    Full Text Available O gênero botânico Clerodendrum pertence à família Lamiaceae e compreende várias espécies ornamentais, Manchas cloróticas e necróticas em folhas de coração-sangrento foram observadas pela primeira vez em um jardim de Piracicaba, SP, associadas à infestação com Brevipalpus phoenicis (Acari: Tenuipalpi [...] dae). Exames de secções de tecidos das lesões foliares ao microscópio eletrônico revelaram ocorrência de efeitos citopáticos do tipo nuclear e concluiu-se que os sintomas eram causados por um vírus transmitido por Brevipalpus (VTB), o qual foi designado de mancha clorótica de Clerodendrum (Clerodendrum Chlorotic Spot Virus- ClCSV). O ClCSV é transmitido mecanicamente de coração-sangrento para coração-sangrento. Em ensaios preliminares foi transmitido por B. phoenicis e mecanicamente para várias outras plantas, além da ocorrência de sua disseminação natural por este ácaro para outras espécies. Visando complementar a caracterização do ClCSV foram feitos estudos sobre alterações anatômicas em folhas de plantas infectadas pelo ClCSV. Foram examinadas secções histológicas de folhas sadias e infectadas pelo ClCSV de C. x speciosum e de outras hospedeiras como Hibiscus schizopetalus, Salvia leucantha, Malvaviscus arboreus e Annona muricata. Constatou-se que o ClCSV causa alterações celulares semelhantes nas diferentes hospedeiras e os sintomas causados por este vírus são similares aos causados por outros vírus transmitidos por Brevipalpus como o vírus da leprose dos citros citoplasmático (Citrus Lepros Virus Cytoplasmic- CiLV-C) e nuclear (Citrus Leprosis Virus Nuclear- CiLV-N), mancha anular do cafeeiro (Coffee Ringspot Virus- CoRSV), mancha anular de Solanum violaefolium (Solanum violaefolium Ringspot Virus- SvRSV) e "Orchid Fleck Vírus" (OFV), representadas por hipertrofia e hiperplasia frequentemente acompanhadas de necrose nos tecidos do parênquima paliçádico e lacunoso. Abstract in english The botanical genus Clerodendrum belongs to the family Lamiaceae and includes several ornamental species. Necrotic and chlorotic spots on leaves of bleeding heart were observed for the first time in a residential garden from Piracicaba Municipality, São Paulo State, Brazil, associated with an infest [...] ation by the flat mite Brevipalpus phoenicis (Acari: Tenuipalpidae). Tissue section analysis of the leaf lesions under transmission electron microscope showed characteristic cytopathic effects of the nuclear type, indicating that the symptoms were caused by a Brevipalpus-transmitted virus (BTV), named Clerodendrum Chlorotic Spot Virus (ClCSV). This virus was shown to be transmitted mechanically and by B. phoenicis to several plant species. In order to complete ClCSV characterization, anatomical changes in the leaf lesions of ClCSV-infected leaves were evaluated. Histological sections of healthy, control leaves and leaf lesions of ClCSV-infected plants of C. x speciosum, Hibiscus schizopetalus, Salvia leucantha, Malvaviscus arboreus and Annona muricata were analyzed. ClCSV infection resulted in similar tissue alterations caused by other BTV as Citrus Leprosies Virus Cytoplasmic (CiLV-C) and Nuclear (CiLV-N), Coffee Ringspot Virus (CoRSV), Solanum violaefolium Ringspot Virus (SvRSV) and Orchid Fleck Virus (OFV). Such symptoms consisted of hypertrophy and hyperplasia frequently followed by necrosis in palisade and spongy parenchyma tissues.

  1. Anatomia de lesões foliares causadas pelo vírus da Mancha Clorótica do Clerodendrum, transmitido pelo ácaro Brevipalpus phoenicis em diferentes espécies Anatomy of leaf lesions caused by Clerodendrum Chlorotic Spot Virus (ClCSV transmitted by the mite Brevipalpus phoenicis in different host species

    Directory of Open Access Journals (Sweden)

    Renata Takassugui Gomes

    2010-12-01

    Full Text Available O gênero botânico Clerodendrum pertence à família Lamiaceae e compreende várias espécies ornamentais, Manchas cloróticas e necróticas em folhas de coração-sangrento foram observadas pela primeira vez em um jardim de Piracicaba, SP, associadas à infestação com Brevipalpus phoenicis (Acari: Tenuipalpidae. Exames de secções de tecidos das lesões foliares ao microscópio eletrônico revelaram ocorrência de efeitos citopáticos do tipo nuclear e concluiu-se que os sintomas eram causados por um vírus transmitido por Brevipalpus (VTB, o qual foi designado de mancha clorótica de Clerodendrum (Clerodendrum Chlorotic Spot Virus- ClCSV. O ClCSV é transmitido mecanicamente de coração-sangrento para coração-sangrento. Em ensaios preliminares foi transmitido por B. phoenicis e mecanicamente para várias outras plantas, além da ocorrência de sua disseminação natural por este ácaro para outras espécies. Visando complementar a caracterização do ClCSV foram feitos estudos sobre alterações anatômicas em folhas de plantas infectadas pelo ClCSV. Foram examinadas secções histológicas de folhas sadias e infectadas pelo ClCSV de C. x speciosum e de outras hospedeiras como Hibiscus schizopetalus, Salvia leucantha, Malvaviscus arboreus e Annona muricata. Constatou-se que o ClCSV causa alterações celulares semelhantes nas diferentes hospedeiras e os sintomas causados por este vírus são similares aos causados por outros vírus transmitidos por Brevipalpus como o vírus da leprose dos citros citoplasmático (Citrus Lepros Virus Cytoplasmic- CiLV-C e nuclear (Citrus Leprosis Virus Nuclear- CiLV-N, mancha anular do cafeeiro (Coffee Ringspot Virus- CoRSV, mancha anular de Solanum violaefolium (Solanum violaefolium Ringspot Virus- SvRSV e "Orchid Fleck Vírus" (OFV, representadas por hipertrofia e hiperplasia frequentemente acompanhadas de necrose nos tecidos do parênquima paliçádico e lacunoso.The botanical genus Clerodendrum belongs to the family Lamiaceae and includes several ornamental species. Necrotic and chlorotic spots on leaves of bleeding heart were observed for the first time in a residential garden from Piracicaba Municipality, São Paulo State, Brazil, associated with an infestation by the flat mite Brevipalpus phoenicis (Acari: Tenuipalpidae. Tissue section analysis of the leaf lesions under transmission electron microscope showed characteristic cytopathic effects of the nuclear type, indicating that the symptoms were caused by a Brevipalpus-transmitted virus (BTV, named Clerodendrum Chlorotic Spot Virus (ClCSV. This virus was shown to be transmitted mechanically and by B. phoenicis to several plant species. In order to complete ClCSV characterization, anatomical changes in the leaf lesions of ClCSV-infected leaves were evaluated. Histological sections of healthy, control leaves and leaf lesions of ClCSV-infected plants of C. x speciosum, Hibiscus schizopetalus, Salvia leucantha, Malvaviscus arboreus and Annona muricata were analyzed. ClCSV infection resulted in similar tissue alterations caused by other BTV as Citrus Leprosies Virus Cytoplasmic (CiLV-C and Nuclear (CiLV-N, Coffee Ringspot Virus (CoRSV, Solanum violaefolium Ringspot Virus (SvRSV and Orchid Fleck Virus (OFV. Such symptoms consisted of hypertrophy and hyperplasia frequently followed by necrosis in palisade and spongy parenchyma tissues.

  2. Poisson's spot with molecules

    International Nuclear Information System (INIS)

    In the Poisson-spot experiment, waves emanating from a source are blocked by a circular obstacle. Due to their positive on-axis interference an image of the source (the Poisson spot) is observed within the geometrical shadow of the obstacle. In this paper we report the observation of Poisson's spot using a beam of neutral deuterium molecules. The wavelength independence and the weak constraints on angular alignment and position of the circular obstacle make Poisson's spot a promising candidate for applications ranging from the study of large molecule diffraction to patterning with molecules.

  3. Nota sobre a molestia de virus do fumo denominada faixa das nervuras

    Directory of Open Access Journals (Sweden)

    A. S. Costa

    1942-02-01

    Full Text Available The present paper deals with the virus disease of the tobacco plant recently described by Kramer & Silberschmidt (12 under the name "faixa das nervuras" (veinbanding. New host plants are added to the list of suscepts, as follows : Nicotiana repanda Willd., JV. angustifolia (*, N. Gossey Domin, N. Sanderae Hort. ex. W. Watson, N. glutinosa L., N. longiflora Cav., N. paniculata L., Lycopersicon esculentum Mill., L. pimpinellifolium Mill., and Cyphomandra betacea Sendt. Datura stramonium L., already studied by Kramer & Silberschmidt (12 and Nicotiana glauca Grah., Nicandra physaloides Gaertn., Solanum nodiflorum (**, S. nigrum L., Lactuca sativa L., Brassica oleracea L., Vigna sinensis Endl. var. Black., and Dolichos lablab L., var. purpurens were found not to be susceptible to the virus. (* This species was received with this name from the Division of Tobacco and Plant Nutrition, U.S.D.A. (** One plant of S. nodiflorum out of 22 which were inoculated gave symptoms of veinbanding. However as no backinoculation was made it can not be stated with certainty if it was truly affected by this virus. The symptomatology on tobacco described by the above mentioned authors agrees as a whole with which has been observed that is : clearing of the veins, chlorotic spotting and veinbanding. In addition it is stated that the white necrotic spotting found associated with the disease on certain tobacco varieties, v. g. Sumatra and Turkish must be attributed to the same virus. On potato plants of the varieties President and Green Mountain the virus causes local streak-like symptoms followed by chlorotic mottling of young leaves. Late symptoms take the form of leaf-drop streak. The virus of "faixa das nervuras" is easily transmissible through the sap giving practically 100% of success. The physical properties of the virus are : tolerance to dilution 1:10.000, thermal death point 54°C. and logevity in vitro 72 hours. No vector of the virus is known. The statement of Kramer & Silberschmidt (12 that the virus of "faixa das nervuras" belongs to the group of the potato virus Y is confirmed. A discussion is given about the position of the virus within the group. Our results do not permit to state that the virus is more related to the veinbanding virus as claimed by these authors. It is also pointed out that the differences found in the litterature between the Y and veinbanding viruses are slight, justifying in the maximum the separation of both, but as strains of the same virus.

  4. Nota sobre a molestia de virus do fumo denominada faixa das nervuras

    Scientific Electronic Library Online (English)

    A. S., Costa; R., Forster.

    1942-02-01

    Full Text Available [...] Abstract in english The present paper deals with the virus disease of the tobacco plant recently described by Kramer & Silberschmidt (12) under the name "faixa das nervuras" (veinbanding). New host plants are added to the list of suscepts, as follows : Nicotiana repanda Willd., JV. angustifolia (*), N. Gossey Domin, N. [...] Sanderae Hort. ex. W. Watson, N. glutinosa L., N. longiflora Cav., N. paniculata L., Lycopersicon esculentum Mill., L. pimpinellifolium Mill., and Cyphomandra betacea Sendt. Datura stramonium L., already studied by Kramer & Silberschmidt (12) and Nicotiana glauca Grah., Nicandra physaloides Gaertn., Solanum nodiflorum (**), S. nigrum L., Lactuca sativa L., Brassica oleracea L., Vigna sinensis Endl. var. Black., and Dolichos lablab L., var. purpurens were found not to be susceptible to the virus. (*) This species was received with this name from the Division of Tobacco and Plant Nutrition, U.S.D.A. (**) One plant of S. nodiflorum out of 22 which were inoculated gave symptoms of veinbanding. However as no backinoculation was made it can not be stated with certainty if it was truly affected by this virus. The symptomatology on tobacco described by the above mentioned authors agrees as a whole with which has been observed that is : clearing of the veins, chlorotic spotting and veinbanding. In addition it is stated that the white necrotic spotting found associated with the disease on certain tobacco varieties, v. g. Sumatra and Turkish must be attributed to the same virus. On potato plants of the varieties President and Green Mountain the virus causes local streak-like symptoms followed by chlorotic mottling of young leaves. Late symptoms take the form of leaf-drop streak. The virus of "faixa das nervuras" is easily transmissible through the sap giving practically 100% of success. The physical properties of the virus are : tolerance to dilution 1:10.000, thermal death point 54°C. and logevity in vitro 72 hours. No vector of the virus is known. The statement of Kramer & Silberschmidt (12) that the virus of "faixa das nervuras" belongs to the group of the potato virus Y is confirmed. A discussion is given about the position of the virus within the group. Our results do not permit to state that the virus is more related to the veinbanding virus as claimed by these authors. It is also pointed out that the differences found in the litterature between the Y and veinbanding viruses are slight, justifying in the maximum the separation of both, but as strains of the same virus.

  5. Identification of Potato Virus Y Strains in Tobacco Crops

    OpenAIRE

    Jelena Zindovi?; Janoš Berenji; Milena Paukovi?; Ivana ?eki?; Aleksandra Bulaji?; Branka Krsti?

    2007-01-01

    Five viruses: Potato Virus Y (PVY), Tomato Spotted Wilt Virus, Cucumber Mosaic Virus, Tobacco Mosaic Virus and Alfalfa Mosaic Virus, of which PVY was predominant, were detected by serological testing of tobacco samples collected from many localities in Vojvodina in 2006. Viruses are the most important pathogens in tobacco and PVY causes considerable economic damages all over the world. A PVY population comprises several different strain groups, strain subgroups and recombinant strains. Among ...

  6. Investigation of likely causes of white patch formation on irradiated WWER fuel rod claddings

    International Nuclear Information System (INIS)

    The information concerning white patches observed on fuel cladding surfaces has been analytically treated. The analysis shows at least three kinds of the white patch appearance: bright white spots which appear to be loose corrosion product deposits disclosing corrosion pits upon spalling; indistinct streaks with separate pronounced spots 1-2 in dia. The spots seem to be thin superficial deposits; light-coloured dense uniform crud distributed over the surface of fuel claddings and fuel assembly jackets. (author)

  7. Models of the Accretion Spots in Magnetic Cataclysmic Variables

    OpenAIRE

    Beuermann, K.

    2003-01-01

    The structure of the near-polar accretion spots on accreting magnetic white dwarfs has been studied theoretically and observationally in numerous papers over the last decade. Detailed treatments are available for the regime of low mass flux, usually termed the bombardment case, and for higher mass fluxes which create a strong shock standing above the photosphere of the white dwarf. No general treatment is so far available for the case of shocks buried deep in the photosphere...

  8. 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 ...

  9. SPOT4 Management Centre

    Science.gov (United States)

    Labrune, Yves; Labbe, X.; Roussel, A.; Vielcanet, P.

    1994-01-01

    In the context of the CNES SPOT4 program CISI is particularly responsible for the development of the SPOT4 Management Centre, part of the SPOT4 ground control system located at CNES Toulouse (France) designed to provide simultaneous control over two satellites. The main operational activities are timed to synchronize with satellite visibilities (ten usable passes per day). The automatic capability of this system is achieved through agenda services (sequence of operations as defined and planned by operator). Therefore, the SPOT4 Management Centre offers limited, efficient and secure human interventions for supervision and decision making. This paper emphasizes the main system characteristics as degree of automation, level of dependability and system parameterization.

  10. Sequence comparison and trasmission of Blackcurrant reversion virus isolates in black, red and white currants with black currant reversion disease and full blossom disease symptoms.

    Czech Academy of Sciences Publication Activity Database

    P?ibylová, Jaroslava; Špak, Josef; Petrzik, Karel; Kubelková, Darina; Špaková, Vlastimila

    2008-01-01

    Ro?. 121, ?. 1 (2008), s. 67-75. ISSN 0929-1873 R&D Projects: GA MŠk 1P05OC051 Institutional research plan: CEZ:AV0Z50510513 Keywords : plant diseases * reversion virus * Sequence comparison Subject RIV: EE - Microbiology, Virology Impact factor: 1.648, year: 2008

  11. Rapid Detection of the Philippine Isolate of Infectious Hypodermal and Hematopoietic Necrosis Virus (IHHNV in Shrimp, Penaeus monodon Using Loop-Mediated Isothermal Amplification (LAMP

    Directory of Open Access Journals (Sweden)

    Christopher Marlowe A. Caipang

    2012-06-01

    Full Text Available The aim of this study was to standardize a Loop-Mediated isothermal Amplification (LAMP assay for the detection of the Philippine isolate of Infectious Hypodermal and Hematopoietic Necrosis Virus (IHHNV in postlarvae of shrimp, Penaeus monodon. The assay was optimized at an incubation time of 1 h at 63oC. The assay was highly specific for IHHNV and did not cross-react with other shrimp viruses including Hepatopancreatic Parvovirus (HPV, Monodon Baculovirus (MBV and White Spot Syndrome Virus (WSSV. The limit of detection of the IHHNV using the LAMP assay was 10 pg of DNA/mL or 10 fg of the genomic DNA per LAMP reaction and was 10 times more sensitive than conventional PCR in detecting the viral pathogen from infected samples. These results demonstrated that LAMP is a simple and sensitive diagnostic technique that has potential application for routine detection of IHHNV infections in shrimp hatcheries in the Philippines.

  12. The Little Red Spot: Closest View Yet

    Science.gov (United States)

    2007-01-01

    This is a mosaic of three New Horizons images of Jupiter's Little Red Spot, taken with the spacecraft's Long Range Reconnaissance Imager (LORRI) camera at 17:41 Universal Time on February 26 from a range of 3.5 million kilometers (2.1 million miles). The image scale is 17 kilometers (11 miles) per pixel, and the area covered measures 33,000 kilometers (20,000 miles) from top to bottom, two and one-half times the diameter of Earth. The Little Red Spot, a smaller cousin of the famous Great Red Spot, formed in the past decade from the merger of three smaller Jovian storms, and is now the second-largest storm on Jupiter. About a year ago its color, formerly white, changed to a reddish shade similar to the Great Red Spot, perhaps because it is now powerful enough to dredge up reddish material from deeper inside Jupiter. These are the most detailed images ever taken of the Little Red Spot since its formation, and will be combined with even sharper images taken by New Horizons 10 hours later to map circulation patterns around and within the storm. LORRI took the images as the Sun was about to set on the Little Red Spot. The LORRI camera was designed to look at Pluto, where sunlight is much fainter than it is at Jupiter, so the images would have been overexposed if LORRI had looked at the storm when it was illuminated by the noonday Sun. The dim evening illumination helped the LORRI camera obtain well-exposed images. The New Horizons team used predictions made by amateur astronomers in 2006, based on their observations of the motion of the Little Red Spot with backyard telescopes, to help them accurately point LORRI at the storm. These are among a handful of Jupiter system images already returned by New Horizons during its close approach to Jupiter. Most of the data being gathered by the spacecraft are stored onboard and will be downlinked to Earth during March and April 2007.

  13. Detection of bluetongue virus RNA in field-collected Culicoides spp. (Diptera: Ceratopogonidae) following the discovery of bluetongue virus serotype 1 in white-tailed deer and cattle in Louisiana

    Science.gov (United States)

    In November 2004, bluetongue virus (family Reoviridae, genus Orbivirus, BTV) serotype 1 (BTV-1) was detected for the first time in the United States from a hunter-killed deer in St. Mary Parish, LA. In 2005, sera surveys were conducted on three cattle farms near the area where the deer was found, an...

  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. Monitoreo de Tagosodes Orizicolus M. e incidencia del virus de la hoja blanca "VHB" en el cultivo de arroz en calabozo, estado guárico, Venezuela / Monitoring of sogata Tagosodes Orizicolus M. and incidence of white leaf virus "WLV" in rice in calabozo, guárico, Venezuela

    Scientific Electronic Library Online (English)

    Luis E, Vivas C; Dilcia, Astudillo; Judith, Poleo.

    2009-12-01

    Full Text Available Se presenta la información sobre la incidencia del virus de la hoja blanca (VHB) y poblaciones del insecto sogata, Tagosodes orizicolus (Muir) 1926, ejecutada en 10 parcelas comerciales del sistema de riego río Guárico (S.R.R.G.), el cual se dividió en tres (3) sectores durante la época de sequía en [...] tre los años 2003 y 2007. Se presenta la información del VHB en 3 parcelas en las épocas de sequía y lluvias. El objetivo del trabajo fue: Evaluar la incidencia en plantas de arroz del VHB y las poblaciones del insecto vector de la enfermedad en siembras comerciales del S.R.R.G. El porcentaje de plantas afectadas por el virus de la hoja blanca alcanzo niveles muy bajos, comprendidos entre 0,56% y 1,08% y las poblaciones del insecto resultaron en promedio de 3,90 individuos por pase doble de malla Abstract in english The incidence of white leaf virus (WLV) and insect population density of sogata, Tagosodes orizicolus (Muir) 1926, was studied in 10 commercial plots of rice located in the Rio Guárico Irrigation System (RG.I.S). The area under the study was divided into three (3) sections during the dry season from [...] 2003 to 2007. The incidence of WLV also was registered in three commercial plots of rice, both in the dry and rainy season. The objective of this study was the evaluation of WLV effect on rice plants as well as the effect of sogata population density as a vector of the disease in commercial plantings (R.G.I.S). The percentage of plants affected by the virus was very low, ranging between 0.56% and 1.08%, and the insect population density of sogata averaged 3.90 individuals per insect net (double pass)

  16. Development and evaluation of a loop-mediated isothermal amplification assay for rapid detection of lymphocystis disease virus.

    Science.gov (United States)

    Li, Qiong; Yue, Zhiqin; Liu, Hong; Liang, Chengzhu; Zheng, Xiaolong; Zhao, Yuran; Chen, Xiao; Xiao, Xizhi; Chen, Changfu

    2010-02-01

    A loop-mediated isothermal amplification (LAMP) assay was developed for the detection of lymphocystis disease virus (LCDV). A set of five specific primers, two inner and two outer primers and a loop primer, were designed on the basis of the major capsid protein gene of LCDV. The reaction time and temperatures were optimized for 60 min at 63 degrees C, respectively. LAMP amplification products were detected by a ladder-like appearance on agarose gel electrophoresis or a naked-eye inspection of a color change in the reaction tube by addition of SYBR Green I. The assay was specific for LCDV, and there was no cross-reactivity with white spot syndrome virus (WSSV) or six other Iridoviridae viruses (epizootic hematopoietic necrosis virus, EHNV; tiger frog virus, TFV; Bohle iridovirus, BIV; soft-shelled turtle iridovirus, STIV; infectious spleen and kidney necrosis virus, ISKNV; red sea bream iridovirus, RSIV). The detection limit of the LAMP assay was 15 fg, which was similar to that of real-time quantitative polymerase chain reaction (PCR) and 10-fold higher than the conventional PCR. The LAMP assay was evaluated using 109 clinical samples, and the results indicated the suitability and simplicity of the test as a rapid, field diagnostic tool for detection of LCDV. The LCDV LAMP assay has potential for early diagnosis of LCDV infection. PMID:19891986

  17. Host defense against DNA virus infection in shrimp is mediated by the siRNA pathway.

    Science.gov (United States)

    Huang, Tianzhi; Zhang, Xiaobo

    2013-01-01

    The RNA interference (RNAi) system of eukaryotes using siRNAs has been documented as an immune response against invasion by RNA viruses. However, whether the siRNA pathway can be triggered by the infection with DNA viruses in animals remains to be investigated. In the present study, we show that Marsupenaeus japonicus shrimp can generate an antiviral siRNA (vp28-siRNA) in response to infection by a double-stranded DNA virus, white spot syndrome virus (WSSV). After challenging with WSSV, vp28-siRNA is detected in all the WSSV-infected organs and tissues of shrimp as early as 24 h postinfection (p.i.). The results indicate that the host Dicer2 and Ago2 proteins are required for the biogenesis and function of vp28-siRNA, respectively. We show further that vp28-siRNA predominates in the cytoplasm of shrimp hemocytes at 48 h p.i. Knockdown of Dicer2 by special siRNA or inhibition of vp28-siRNA with locked nucleic acid antisense oligonucleotides both lead to a significant increase in WSSV copy number at 24-48 h p.i. Our study highlights a novel aspect of the siRNA pathway in the immune response of animals against infection by DNA viruses. PMID:23065729

  18. Inhibition of Taura syndrome virus replication in Litopenaeus vannamei through silencing the LvRab7 gene using double-stranded RNA.

    Science.gov (United States)

    Ongvarrasopone, Chalermporn; Saejia, Pipop; Chanasakulniyom, Mayuree; Panyim, Sakol

    2011-07-01

    Taura syndrome virus (TSV) is a major cause of high mortality in Pacific white shrimp (Litopenaeus vannamei, Lv). Previously, silencing of Penaeus monodon Rab7 (PmRab7) by injecting double-stranded RNA corresponding to PmRab7 (dsRNA-PmRab7) prevented white spot syndrome virus or yellow head virus infection. Rab7 is proposed to be involved in intracellular trafficking of the viruses. This study aimed to investigate whether knockdown of Rab7 in L. vannamei by dsRNA-PmRab7 could inhibit replication of TSV. RNA interference (RNAi) technology using dsRNA targeting the LvRab7 gene was used to silence the mRNA expression of LvRab7. The silencing of the LvRab7 gene inhibited TSV replication dramatically when compared to groups receiving dsRNA-GFP or NaCl. This is the first demonstration that dsRNA targeting the endogenous shrimp gene LvRab7 strongly reduces TSV replication. It provides further evidence that LvRab7 is involved in the endosomal trafficking pathway of viruses infecting penaeid shrimp. PMID:21347841

  19. Spotting a fake

    International Nuclear Information System (INIS)

    Diamonds are highly prized for their dazzling appearance and hardness, but would you be able to spot one that had been created in the laboratory? Simon Lawson describes how physics-based techniques can distinguish between natural and synthetic stones. For the last 50 years or so we have been able to make synthetic diamonds that replicate the superlative physical and chemical properties of natural diamonds, and these are used largely for industrial applications. But in the mind of the consumer, there is far more to a diamond than its hardness or brilliance. Research commissioned by the Diamond Trading Company (DTC) has shown that 94% of women surveyed prefer natural diamonds over synthetic ones as a symbol of love, possibly as a result of the immense age of natural stones. One of the key research activities at the DTC is therefore to ensure that synthetic diamonds can be spotted easily. (U.K.)

  20. Spot- Zombie Filtering System

    Directory of Open Access Journals (Sweden)

    Arathy Rajagopal

    2014-01-01

    Full Text Available A major security challenge on the Internet is the existence of the large number of compromised machines. Such machines have been increasingly used to launch various security attacks including spamming and spreading malware, DDoS, and identity theft. These compromised machines are called “Zombies”. In general E-mail applications and providers uses spam filters to filter the spam messages. Spam filtering is a technique for discriminating the genuine message from the spam messages. The attackers send the spam messages to the targeted machine by exalting the filters, which causes the increase in false positives and false negatives. We develop an effective spam zombie detection system named SPOT by monitoring outgoing messages of a network. SPOT focuses on the number of outgoing messages that are originated or forwarded by each computer on a network to identify the presence of Zombies. SPOT is designed based on a powerful statistical tool called Sequential Probability Ratio Test, which has bounded false positive and false negative error rates.

  1. WHITE DWARFS

    OpenAIRE

    Horn, H.

    1980-01-01

    The thermal properties and neutrino emissivity of matter in the dense, degenerate cores of white dwarfs, together with efficient heat transport through the high-density envelopes of these stars, affect the theoretical white dwarf luminosity function. The most recent observational data show departures from the results of elementary cooling theory at both high and low luminosities. Extreme UV and soft x-ray observations from space show the necessity of neutrino cooling for white dwarfs, have be...

  2. Avaliação de produtos químicos comerciais, in vitro e in vivo, no controle da doença foliar, mancha branca do milho, causada por Pantoea ananatis Evaluation of commercial chemical products, in vitro and in vivo in the control of foliar disease, maize white spot, caused by Pantoea ananais

    Directory of Open Access Journals (Sweden)

    Cleide Aparecida Bomfeti

    2007-03-01

    Full Text Available Uma bactéria identificada como Pantoea ananatis foi recentemente isolada de lesões jovens da doença mancha branca do milho de plantas naturalmente infectadas. Esta bateria reproduziu sintomas semelhantes aos da doença quando inoculada em plantas de milho em casa de vegetação. Estudos anteriores realizados por outros autores demonstraram que o controle desta doença em condições de campo foi obtido pelo uso de fungicidas, principalmente o Mancozeb, nas fases iniciais de seu desenvolvimento. O objetivo deste estudo foi avaliar a freqüência de isolamento da bactéria P. ananatis a partir de plantas infectadas coletadas na região de Londrina, Estado do Paraná, e reproduzir sintomas da doença através de inoculações artificiais em plantas de milho em casa de vegetação. Utilizando os produtos químicos testados anteriormente por outros autores para o controle desta doença a campo, foi também objetivo deste trabalho avaliar o potencial destes produtos na inibição da bactéria tanto em condições de laboratório como em condições de infecção natural. Os resultados mostraram que P. ananatis foi isolada em 40% das lesões jovens coletadas a campo e quando inoculada em casa de vegetação sob condições controladas reproduziu sintomas semelhantes aos observados a campo. Entre os produtos químicos testados, o fungicida Mancozeb mostrou-se eficiente no controle da doença a campo, em concordância com os relatos anteriores. Este produto inibiu completamente o crescimento da bactéria em laboratório, explicando os resultados obtidos a campo. Os demais produtos não foram eficientes no controle a campo e eles também não inibiram a bactéria em laboratório. Estes resultados representam evidências adicionais de que a bactéria P. ananatis é o agente causal da doença mancha branca do milho.A bacterium identified as Pantoea ananatis was recently isolated from young lesions of the disease maize white spot from infected plants in the field. This bacterium produced similar symptoms of the disease when inoculated on maize plants in greenhouse. Previous studies carried out by other authors demonstrated that the control of this disease in field conditions has been achieved through the use of fungicides mainly with the Mancozeb in the initial phases of its development. The aim of this study was to evaluate the frequency of isolation of P. ananatis from infected plants in the region of "Londrina, Paraná State" and to reproduce symptoms of the disease through artificial inoculation of maize plants in greenhouse. Using the chemical products tested previously by other authors for the control of this disease in the field, it was also objective of this work to evaluate the potential of these products to inhibit the bacterium, at both in laboratory and in condition of natural infection. The results showed that P. ananatis was successfully isolated from 40% of the young lesions collected in the field and when inoculated in greenhouse (controlled conditions, it reproduced symptoms similar to what was observed in the field. Among the tested chemical products, the fungicide Mancozeb was the best in the control of the disease under field conditions, in agreement with previous reports. This product inhibited completely the bacterium in laboratory, explaining the results found in the field. The others products were not efficient to control the disease and they did not also inhibit the bacterium in laboratory. The results represent additional evidences that the bacterium P. ananatis is the causal agent of the maize white spot disease.

  3. Avaliação de produtos químicos comerciais, in vitro e in vivo, no controle da doença foliar, mancha branca do milho, causada por Pantoea ananatis / Evaluation of commercial chemical products, in vitro and in vivo in the control of foliar disease, maize white spot, caused by Pantoea ananais

    Scientific Electronic Library Online (English)

    Cleide Aparecida, Bomfeti; Walter Fernandes, Meirelles; Edneia Aparecida, Souza-Paccola; Carlos Roberto, Casela; Alexandre da Silva, Ferreira; Ivanildo Evódio, Marriel; Luzia Doretto, Paccola-Meirelles.

    2007-03-01

    Full Text Available Uma bactéria identificada como Pantoea ananatis foi recentemente isolada de lesões jovens da doença mancha branca do milho de plantas naturalmente infectadas. Esta bateria reproduziu sintomas semelhantes aos da doença quando inoculada em plantas de milho em casa de vegetação. Estudos anteriores real [...] izados por outros autores demonstraram que o controle desta doença em condições de campo foi obtido pelo uso de fungicidas, principalmente o Mancozeb, nas fases iniciais de seu desenvolvimento. O objetivo deste estudo foi avaliar a freqüência de isolamento da bactéria P. ananatis a partir de plantas infectadas coletadas na região de Londrina, Estado do Paraná, e reproduzir sintomas da doença através de inoculações artificiais em plantas de milho em casa de vegetação. Utilizando os produtos químicos testados anteriormente por outros autores para o controle desta doença a campo, foi também objetivo deste trabalho avaliar o potencial destes produtos na inibição da bactéria tanto em condições de laboratório como em condições de infecção natural. Os resultados mostraram que P. ananatis foi isolada em 40% das lesões jovens coletadas a campo e quando inoculada em casa de vegetação sob condições controladas reproduziu sintomas semelhantes aos observados a campo. Entre os produtos químicos testados, o fungicida Mancozeb mostrou-se eficiente no controle da doença a campo, em concordância com os relatos anteriores. Este produto inibiu completamente o crescimento da bactéria em laboratório, explicando os resultados obtidos a campo. Os demais produtos não foram eficientes no controle a campo e eles também não inibiram a bactéria em laboratório. Estes resultados representam evidências adicionais de que a bactéria P. ananatis é o agente causal da doença mancha branca do milho. Abstract in english A bacterium identified as Pantoea ananatis was recently isolated from young lesions of the disease maize white spot from infected plants in the field. This bacterium produced similar symptoms of the disease when inoculated on maize plants in greenhouse. Previous studies carried out by other authors [...] demonstrated that the control of this disease in field conditions has been achieved through the use of fungicides mainly with the Mancozeb in the initial phases of its development. The aim of this study was to evaluate the frequency of isolation of P. ananatis from infected plants in the region of "Londrina, Paraná State" and to reproduce symptoms of the disease through artificial inoculation of maize plants in greenhouse. Using the chemical products tested previously by other authors for the control of this disease in the field, it was also objective of this work to evaluate the potential of these products to inhibit the bacterium, at both in laboratory and in condition of natural infection. The results showed that P. ananatis was successfully isolated from 40% of the young lesions collected in the field and when inoculated in greenhouse (controlled conditions), it reproduced symptoms similar to what was observed in the field. Among the tested chemical products, the fungicide Mancozeb was the best in the control of the disease under field conditions, in agreement with previous reports. This product inhibited completely the bacterium in laboratory, explaining the results found in the field. The others products were not efficient to control the disease and they did not also inhibit the bacterium in laboratory. The results represent additional evidences that the bacterium P. ananatis is the causal agent of the maize white spot disease.

  4. Influence of temperature on Mx gene expression profiles and the protection of sevenband grouper, Epinephelus septemfasciatus, against red-spotted grouper nervous necrosis virus (RGNNV) infection after poly (I:C) injection.

    Science.gov (United States)

    Thanasaksiri, Kittipong; Sakai, Nichika; Yamashita, Hirofumi; Hirono, Ikuo; Kondo, Hidehiro

    2014-10-01

    Influence of temperature on the susceptibility of fish against virus infection has been studied for a decade. Recent reports have been shown the effects of rearing temperatures on the fish immune system against virus infection. However, the roles of temperature in regulation of type I interferon (IFN) system has not yet been investigated. Thus, the effects of temperature on type I IFN response were investigated in this study using poly (I:C) injection in sevenband grouper and Mx gene was used as a marker for type I IFN expression. Quantitative real-time PCR (qPCR) result showed that Mx expression profiles were moderately different between temperatures. The highly up-regulated Mx transcripts at 3 h post injection (hpi) were observed in high temperatures (25 °C and 30 °C) but not in low temperatures (15 °C and 20 °C). Meanwhile, low temperatures (15 °C and 20 °C) could detect the highly up-regulated Mx transcripts at 24 hpi. Expression of Mx transcripts was also observed at 72 hpi at 15 °C. Poly (I:C)-injected fish were challenged with RGNNV after 72 and 168 hpi. At 72 hpi, 100% of fish survived at all temperatures, whereas 95% survival rate was observed at 168 hpi at 25 °C during 14 days of observation. To further verify the duration period of an antiviral state at different temperatures, qPCR and endpoint dilution assay were used to quantify the number of virus in fish challenged with RGNNV. The reduction of viral copy numbers and viral titers could be observed at 72 and 168 hpi. However, high viral copy numbers and viral titers could be detected at 168 hpi at 30 °C. These results demonstrate that temperatures influenced on the Mx expression profiles and the duration period of an antiviral state efficiently interfered with virus replication at different temperatures. PMID:25107694

  5. Models of the Accretion Spots in Magnetic Cataclysmic Variables

    CERN Document Server

    Beuermann, K

    2003-01-01

    The structure of the near-polar accretion spots on accreting magnetic white dwarfs has been studied theoretically and observationally in numerous papers over the last decade. Detailed treatments are available for the regime of low mass flux, usually termed the bombardment case, and for higher mass fluxes which create a strong shock standing above the photosphere of the white dwarf. No general treatment is so far available for the case of shocks buried deep in the photosphere. I review the theoretical foundations, present some applications of theory, and discuss in short the open questions which still need to be addressed.

  6. Presença do vírus da síndrome da mancha branca em crustáceos decápodes silvestres em lagoas costeiras no Sul do Brasil Presence of the white spot syndrome virus (WSSV) in wild decapods crustaceans in coastal lagoons in southern Brazil

    OpenAIRE

    S.W. Costa; A.P.M. Fraga; A.S. Zamparetti; M. R. F. MARQUES; E.R. Andreatta

    2012-01-01

    A presença do vírus da síndrome da mancha branca (em inglês WSSV) nas principais espécies de camarões, siris e caranguejos de cinco lagoas que recebem o efluente de fazendas afetadas pela enfermidade foi detectada por nested PCR, e inclusões virais nos camarões por histologia. Pela nested PCR encontrou-se a presença de WSSV em 13 de 16 (81,2%) amostras de camarões da espécie Farfantepenaeus paulensis, em 13 de 14 (92,8%) de Litopenaeus schmitti, em uma de duas de Farfantepenaeus brasiliensis ...

  7. Top 10 plant viruses in molecular plant pathology.

    Science.gov (United States)

    Scholthof, Karen-Beth G; Adkins, Scott; Czosnek, Henryk; Palukaitis, Peter; Jacquot, Emmanuel; Hohn, Thomas; Hohn, Barbara; Saunders, Keith; Candresse, Thierry; Ahlquist, Paul; Hemenway, Cynthia; Foster, Gary D

    2011-12-01

    Many scientists, if not all, feel that their particular plant virus should appear in any list of the most important plant viruses. However, to our knowledge, no such list exists. The aim of this review was to survey all plant virologists with an association with Molecular Plant Pathology and ask them to nominate which plant viruses they would place in a 'Top 10' based on scientific/economic importance. The survey generated more than 250 votes from the international community, and allowed the generation of a Top 10 plant virus list for Molecular Plant Pathology. The Top 10 list includes, in rank order, (1) Tobacco mosaic virus, (2) Tomato spotted wilt virus, (3) Tomato yellow leaf curl virus, (4) Cucumber mosaic virus, (5) Potato virus Y, (6) Cauliflower mosaic virus, (7) African cassava mosaic virus, (8) Plum pox virus, (9) Brome mosaic virus and (10) Potato virus X, with honourable mentions for viruses just missing out on the Top 10, including Citrus tristeza virus, Barley yellow dwarf virus, Potato leafroll virus and Tomato bushy stunt virus. This review article presents a short review on each virus of the Top 10 list and its importance, with the intent of initiating discussion and debate amongst the plant virology community, as well as laying down a benchmark, as it will be interesting to see in future years how perceptions change and which viruses enter and leave the Top 10. PMID:22017770

  8. Virus fitopatógenos en insectos asociados al ajo / Plant viruses in garlic-associated insects

    Scientific Electronic Library Online (English)

    Octavio Gustavo, García-Rodríguez; Luis, Pérez-Moreno; Martha Juana, Navarro-León; Manuel Darío, Salas-Araiza; Oscar Alejandro, Martínez-Jaime; Ma. Fabiola, León-Galván; Héctor Gordon, Núñez-Palenius.

    2014-08-01

    Full Text Available La propagación vegetativa del ajo (Allium sativum) es la principal vía de transmisión de virus para este cultivo. No obstante, no se debe descartar la dispersión por insectos vectores. El objetivo del presente estudio fue detectar, mediante la prueba de ELISA, la presencia de virus en insectos colec [...] tados en plantas de ajo. El experimento se llevó a cabo durante el ciclo Otoño-Invierno 2008-2009. La toma de muestras de insectos se realizó en tres fechas después de la siembra. La identificación de las especies de insectos se realizó utilizando un estereomicroscopio Zeiss de 30X y claves taxonómicas. La detección de virus fue mediante la técnica de DAS-ELISA para los potyvirus Virus del rayado amarillo del puerro (Leek yellow spot virus: LYSV) y Virus del enanismo amarillo de la cebolla (Onion yellow dwarf virus: OYDV); los carlavirus Virus latente común del ajo (Garlic common latent virus: GCLV) y Virus latente del chalote (Shallot latent virus: SLV); y el tospovirus Virus del manchado amarillo del iris (Iris yellow spot virus: IYSV). Se identificaron 19 especies de insectos, de los que destacaron Thrips tabaci Lindeman, como positivo en 18 muestras para GCLV y dos muestras para IYSV, y Collops quadrimaculatus, como positivo en una muestra para GCLV. Abstract in english Vegetative propagation of garlic (Allium sativum) is the main route of virus transmission for this crop. However, insect-vector spread should not be ruled out. The aim of this study was to detect, by means of the ELISA test, the presence of five viruses in insects collected in garlic plants. The exp [...] eriment was conducted during the fall-winter (2008-2009) cycle. Insect samples were taken on three dates: 45, 110 and 140 days after garlic sowing. Insect species identification was performed using a Zeiss (30X) stereomicroscope and the O'Brien and Wilson (1985) and Mound and Kibby (1998) taxonomy keys. The serology test for virus detection was the DAS-ELISA technique. Coat-protein virus antibodies were applied for the potyviruses: Leek yellow spot virus (LYSV) and Onion yellow dwarf virus (OYDV); for the carlaviruses: Garlic common latent virus (GCLV) and Shallot latent virus (SLV); and for the tospovirus: Iris yellow spot virus (IYSV). Of the 19 insect species identified, Thrips tabaci Lindeman tested positive in 18 samples for GCLV and in 2 samples for IYSV, and Collops quadrimaculatus tested positive in one sample for GCLV.

  9. Advances in spot curing technology

    International Nuclear Information System (INIS)

    A brief review of spot curing technology was presented. The process which a spot of energy of a specific wavelength bandwidth and irradiance is used to cause a coating, encapsulant or adhesive to change from a liquid to a solid state

  10. Japanese Spotted Fever, South Korea

    OpenAIRE

    Chung, Moon-hyun; Lee, Seung-hyun; Kim, Mi-jeong; Lee, Jung-hee; Kim, Eun-sil; Lee, Jin-soo; Kim, Mee-kyung; Park, Mi-yeoun; Kang, Jae-seung

    2006-01-01

    We describe the first case of Japanese spotted fever and the first isolate of spotted fever group rickettsia from a patient in South Korea. The isolated rickettsia from the patient was identified as Rickettsia japonica by analysis of the nucleotide sequences of 16S rRNA, gltA, ompA, ompB, and sca4 genes.

  11. A New Age Approach to the Management of Tomato Spotted Wilt? Effects of Plant Essential Oils and Particle Films on Tomato Spotted Wilt in Tomatoes

    Science.gov (United States)

    Because the thrips-vectored Tomato spotted wilt virus is a limiting factor in tomato production in the southern USA, we are investigating novel control methods that would be effective and environmentally non-disruptive. In laboratory choice tests, we found that three plant essential oils, geraniol, ...

  12. Programming balanced optical beam splitters in white paint

    OpenAIRE

    Huisman, S. R.; Huisman, T. J.; Goorden, S. A.; Mosk, A. P.; Pinkse, P. W. H.

    2014-01-01

    Wavefront shaping allows for ultimate control of light propagation in multiple-scattering media by adaptive manipulation of incident waves. We shine two separate wavefront-shaped beams on a layer of dry white paint to create two enhanced output speckle spots of equal intensity. We experimentally confirm by interference measurements that the output speckle spots are almost correlated like the two outputs of an ideal balanced beam splitter. The observed deviations from the pha...

  13. Control of aphid-vectored and thrips-borne virus spread in lily, tulip, iris and dahlia by sprays of mineral oil, polydimethylsiloxane and pyrethroid insecticide in the field

    OpenAIRE

    Asjes, J.; Blom-Barnhoorn, G.J.

    2001-01-01

    In this study control of spread by insect vectors of non-persistent Lily symptomless virus and Lily mottle virus in lily, Tulip breaking virus in tulip, Iris mild mosaic virus, Narcissus latent virus and Iris severe mosaic virus in bulbous iris, and semi-persistent Dahlia mosaic virus and persistent Tomato spotted wilt virus in dahlia has been evaluated with weekly sprays of mineral oil, beta-pinene emulsion, polydimethylsiloxane emulsions and pyrethroid insecticide. In lily, beta-pinene in '...

  14. Characterization of host microRNAs that respond to DNA virus infection in a crustacean

    Directory of Open Access Journals (Sweden)

    Huang Tianzhi

    2012-04-01

    Full Text Available Abstract Background MicroRNAs (miRNAs are key posttranscriptional regulators of gene expression that are implicated in many processes of eukaryotic cells. It is known that the expression profiles of host miRNAs can be reshaped by viruses. However, a systematic investigation of marine invertebrate miRNAs that respond to virus infection has not yet been performed. Results In this study, the shrimp Marsupenaeus japonicus was challenged by white spot syndrome virus (WSSV. Small RNA sequencing of WSSV-infected shrimp at different time post-infection (0, 6, 24 and 48?h identified 63 host miRNAs, 48 of which were conserved in other animals, representing 43 distinct families. Of the identified host miRNAs, 31 were differentially expressed in response to virus infection, of which 25 were up-regulated and six down-regulated. The results were confirmed by northern blots. The TargetScan and miRanda algorithms showed that most target genes of the differentially expressed miRNAs were related to immune responses. Gene ontology analysis revealed that immune signaling pathways were mediated by these miRNAs. Evolutionary analysis showed that three of them, miR-1, miR-7 and miR-34, are highly conserved in shrimp, fruit fly and humans and function in the similar pathways. Conclusions Our study provides the first large-scale characterization of marine invertebrate miRNAs that respond to virus infection. This will help to reveal the molecular events involved in virus-host interactions mediated by miRNAs and their evolution in animals.

  15. White Pelican

    Science.gov (United States)

    The American white pelican is still considered endangered in Alberta, Canada, where the population is increasing but fewer than half of the 20 known historic nesting islands are still in use. The site provides information on this magnificent bird: habitat, general biological data, risk factors, and management. External links to Canadian parks, nonprofit groups, and other species profiles also included.

  16. 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.

  17. Identification of a JAK/STAT pathway receptor domeless from Pacific white shrimp Litopenaeus vannamei.

    Science.gov (United States)

    Yan, Muting; Li, Chaozheng; Su, Ziqi; Liang, Qianhui; Li, Haoyang; Liang, Shizhong; Weng, Shaoping; He, Jianguo; Xu, Xiaopeng

    2015-05-01

    The Janus kinase/signal transducers and activators of transcription (JAK/STAT) signaling pathway was known to participate in dozens of immune responses in organisms. Domeless, first identified in Drosophila melanogaster, is a unique receptor involved in invertebrate JAK/STAT pathway. In this study, a cytokine receptor (LvDOME) was identified in Litopenaeus vannamei. The LvDOME cDNA was 5178bp in length with an Open Reading Frame (ORF) of 4191bp. LvDOME contained two cytokine binding modules (CBMs) and three fibronectin-type-III-like (FNIII) domains, similar to most vertebrate IL-6 receptors. LvDOME was expressed highest in shrimp muscle and could be up-regulated in the late stage of white spot syndrome virus (WSSV) infection. LvDOME could significantly enhance the activity of the WSSV wsv069 gene promoter through acting on the STAT-binding motif, suggesting LvDOME could activate the JAK/STAT pathway. Moreover, knockdown of LvDOME resulted in lower cumulative mortality of shrimps and less WSSV copies, suggesting LvDOME may be hijacked by WSSV to benefit virus replication. To our knowledge, this is the first report on the receptor of JAK/STAT pathway in shrimp. PMID:25659232

  18. Mopeia Virus-related Arenavirus in Natal Multimammate Mice, Morogoro, Tanzania

    DEFF Research Database (Denmark)

    Günther, Stephan; Hoofd, Guy

    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 Morogoro virus, although a large proportion shows specific antibodies.

  19. Polarization from an orbiting spot

    Science.gov (United States)

    Dov?iak, Michal; Karas, Vladimír; Matt, Giorgio

    2007-04-01

    The polarization from a spot orbiting around Schwarzschild and extreme Kerr black holes is studied. We assume different models of local polarization. Firstly, as a toy model we set local polarization vector either normal to the disc plane, or we assume strictly azimuthal direction. Then we examine more realistic situation with a spot arising due to the emission from the primary source above the disc. We employ either Rayleigh single scattering or Compton multiple scattering approximations. Overall flux, degree and angle of polarization integrated over the whole orbit as well as their time dependence during the spot revolution are examined as functions of the observer's inclination angle. The gravitational and Doppler shifts, lensing effect as well as time delays are taken into account.

  20. NCBI nr-aa BLAST: CBRC-CREM-01-1232 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-CREM-01-1232 ref|NP_478006.1| wsv484 [Shrimp ... white spot syndrome virus] gb|AAL33485.1| wsv4 ... 84 [shrimp ... white spot syndrome virus] gb|AAL88879.1| WSSV011 ... [shrimp ... white spot syndrome virus] NP_478006.1 8.9 27% ...

  1. NCBI nr-aa BLAST: CBRC-DNOV-01-2338 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-DNOV-01-2338 ref|NP_477662.1| wsv140 [Shrimp ... white spot syndrome virus] gb|AAL33144.1| wsv1 ... 40 [shrimp ... white spot syndrome virus] gb|AAL89063.1| WSSV195 ... [shrimp ... white spot syndrome virus] NP_477662.1 0.54 32% ...

  2. NCBI nr-aa BLAST: CBRC-OPRI-01-1405 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-OPRI-01-1405 ref|NP_477758.1| wsv236 [Shrimp ... white spot syndrome virus] gb|AAL33240.1| wsv2 ... 36 [shrimp ... white spot syndrome virus] gb|AAL89160.1| WSSV292 ... [shrimp ... white spot syndrome virus] NP_477758.1 2.4 29% ...

  3. NCBI nr-aa BLAST: CBRC-OGAR-01-1331 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-OGAR-01-1331 ref|NP_477761.1| wsv239 [Shrimp ... white spot syndrome virus] gb|AAL33243.1| wsv2 ... 39 [shrimp ... white spot syndrome virus] gb|AAL89163.1| WSSV295 ... [shrimp ... white spot syndrome virus] NP_477761.1 7e-05 36% ...

  4. NCBI nr-aa BLAST: CBRC-EEUR-01-1367 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-EEUR-01-1367 ref|NP_477616.1| wsv094 [Shrimp ... white spot syndrome virus] gb|AAL33098.1| wsv0 ... 94 [shrimp ... white spot syndrome virus] gb|AAL89018.1| WSSV150 ... [shrimp ... white spot syndrome virus] NP_477616.1 1.7 37% ...

  5. Gene : CBRC-OGAR-01-1331 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-OGAR-01-1331 Novel UN D UNKNOWN EXTN_TOBAC 0.004 34% ref|NP_477761.1| wsv239 [Shrimp ... white ... spot syndrome virus] gb|AAL33243.1| wsv239 [shrimp ... white spot syndrome virus] gb|AAL89163.1| WSSV295 ... [shrimp ... white spot syndrome virus] 7e-05 36% MALDVKTKVLTMT ...

  6. HOT SPOT Health Physics codes

    International Nuclear Information System (INIS)

    The newly developed HOT SPOT Health Physics codes allow fast assessment of accidents involving radioactive materials. These codes run on the Hewlett-Packard HP-41 family of hand-held computers, and the software resides on a single 128K minicassette. This compact system is intended for easy field use of the HOT SPOT codes. At present, 13 separate programs exist, ranging from general programs for downwind assessment following the release of radioactive material to more specific programs dealing with the release of plutonium, uranium, or tritium. Other programs estimate the dose commitment from the inhalation of various radionuclides, and estimate the effects of a surface-burst nuclear weapon

  7. Color tunable LED spot lighting

    Science.gov (United States)

    Hoelen, C.; Ansems, J.; Deurenberg, P.; van Duijneveldt, W.; Peeters, M.; Steenbruggen, G.; Treurniet, T.; Valster, A.; ter Weeme, J. W.

    2006-08-01

    A new trend in illumination is to use dynamic light to set or dynamically vary the ambience of a room or office. For this we need color tunable spots that can reliably vary over at least a wide range of color temperatures, and preferably also more saturated colors. LEDs are in principle ideally suited for this application thanks to their nature of emitting light in a relatively narrow band. For color tunable spot lighting based on the concept of mixing RGB LED colors, the key results have been presented before. Limitations of these 3-intrinsic-color mixing systems with high color rendering properties are found in a limited operating temperature range due to wavelength shifts, a limited color temperature range, and a low maximum operating temperature due to a strong flux decrease with increasing temperature. To overcome these limitations, a 3-color R pcGB system with phosphor-converted red (R pc) and a 4-color RAGB system have been investigated. With both systems, a CRI of at least 80 can be maintained over the relevant color temperature range of approximately 2700 K to 6500 K. In this paper we compare these concepts on overall system aspects and report on the performance of prototype spot lamps. The main features of the RAGB and R pcGB spot lamp concepts can be summarized as: 1) The RAGB spot overcomes CRI and gamut shortcomings of RGB light sources and gives much freedom in wavelength selection, but suffers from temperature sensitivity and complex controls; 2) The R pcGB spot overcomes shortcomings concerning CRI and thermal dependence of RGB sources and enables relatively simple controls, but needs an improved overall red efficacy. With both color concepts, prototype spot lamps have been built. The amber to red emitting nitridosilicate-based phosphors can be wavelength-tuned for optimal performance, which is found at a peak emission around 610 nm for high color quality systems. This results in a simple and very robust system with good color consistency. For the RAGB system, a spot lamp has been developed, consisting of a 4-chip light source, an optical system with mixing rod that provides color homogenization and beam shaping, and an electronic drive and control unit based on temperature feed forward. Flux- and color-rendering performance can be tuned according to the application requirements.

  8. 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.

  9. Is this Red Spot the Blue Spot (locus ceruleum)?

    International Nuclear Information System (INIS)

    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.

  10. Is this Red Spot the Blue Spot (locus ceruleus)?

    OpenAIRE

    Choe, Wonsick; Lee, YuKyung; Lee, Minkyung; Hwang, Kyung Hoon

    2010-01-01

    The authors report brain images of 18F-FDG-PET in a case of schizophrenia. The images showed strikingly increased bilateral uptake in the locus ceruleus. The locus ceruleus is called the blue spot and known to be a center of the norepinephrinergic system.

  11. Rocky Mountain Spotted Fever, Panama

    OpenAIRE

    Estripeaut, Dora; Aramburú, María Gabriela; Sáez-Llorens, Xavier; Thompson, Herbert A; Dasch, Gregory A.; Paddock, Christopher D; Zaki, Sherif; Eremeeva, Marina E.

    2007-01-01

    We describe a fatal pediatric case of Rocky Mountain spotted fever in Panama, the first, to our knowledge, since the 1950s. Diagnosis was established by immunohistochemistry, PCR, and isolation of Rickettsia rickettsii from postmortem tissues. Molecular typing demonstrated strong relatedness of the isolate to strains of R. rickettsii from Central and South America.

  12. Tomato chocolàte virus: a new plant virus infecting tomato and a proposed member of the genus Torradovirus

    OpenAIRE

    Verbeek, M.; Dullemans, A. M.; Heuvel, J. F. J. M.; Maris, P. C.; Vlugt, 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) and Tomato marchitez virus (ToMarV), both members of the genus Torradovirus. The name tomato chocolàte virus is proposed for this new torradovirus. M. Verbeek and A. Dullemans contributed to this re...

  13. An online real time ultrasonic NDT system for the quality control of spot welding in the automotive industry

    International Nuclear Information System (INIS)

    Resistance spot welding is the main joining technique used for the fabrication of body-in-white structures in the automotive industry. The quality of the welds depends on the profile of the spot welding electrode cap. The increased use of zinc coated steel in the industry increases wear rate of the caps, making quality control more difficult. This paper presents a novel online real time ultrasonic NDE system for resistance spot welding which evaluates every weld as it is formed. SEM results are presented to show the alloying of the electrode caps.

  14. RASPBERRY MOSAIC DISEASE, AN EXPANDING VIRUS UNIVERSE

    Science.gov (United States)

    Raspberries are susceptible to multiple viruses, some of unknown etiology. Four of them are spread almost exclusively by aphids in the genus Amphorophora and make up the raspberry mosaic complex, which consists of Black raspberry necrosis, Rubus yellow net, Raspberry leaf spot, and Raspberry leaf m...

  15. The genome of Oryctes rhinoceros nudivirus provides novel insight into the evolution of nuclear arthropod-specific large circular double-stranded DNA viruses.

    Science.gov (United States)

    Wang, Yongjie; Bininda-Emonds, Olaf R P; van Oers, Monique M; Vlak, Just M; Jehle, Johannes A

    2011-06-01

    The Oryctes rhinoceros nudivirus (OrNV) is a dsDNA virus with enveloped, rod-shaped virions. Its genome is 127,615 bp in size and contains 139 predicted protein-coding open reading frames (ORFs). In-depth genome sequence comparisons revealed a varying number of shared gene homologues, not only with other nudiviruses (NVs) and baculoviruses, but also with other arthropod-specific large dsDNA viruses, including the so-called Monodon baculovirus (MBV), the salivary gland hypertrophy viruses (SGHVs) and white spot syndrome virus (WSSV). Nudivirus genomes contain 20 baculovirus core gene homologues associated with transcription (p47, lef-8, lef-9, lef-4, vlf-1, and lef-5), replication (dnapol and helicase), virus structure (p74, pif-1, pif-2, pif-3, 19kda/pif-4, odv-e56/pif-5, vp91, vp39, and 38K), and unknown functions (ac68, ac81, and p33). Most strikingly, a set of homologous genes involved in peroral infection (p74, pif-1, pif-2, and pif-3) are common to baculoviruses, nudiviruses, SGHVs, and WSSV indicating an ancestral mode of infection in these highly diverged viruses. A gene similar to polyhedrin/granulin encoding the baculovirus occlusion body protein was identified in non-occluded NVs and in Musca domestica SGHV evoking the question of the evolutionary origin of the baculovirus polyhedrin/granulin gene. Based on gene homologies, we further propose that the shrimp MBV is an occluded member of the nudiviruses. We conclude that baculoviruses, NVs and the shrimp MBV, the SGHVs and WSSV share the significant number of conserved genetic functions, which may point to a common ancestry of these viruses. PMID:21380757

  16. Improvements on analytic modelling of stellar spots

    CERN Document Server

    Montalto, M; Oshagh, M; Boisse, I; Bruno, G; Santos, N C

    2014-01-01

    In this work we present the solution of the stellar spot problem using the Kelvin-Stokes theorem. Our result is applicable for any given location and dimension of the spots on the stellar surface. We present explicitely the result up to the second degree in the limb darkening law. This technique can be used to calculate very efficiently mutual photometric effects produced by eclipsing bodies occulting stellar spots and to construct complex spot shapes.

  17. The spot market and the spot price: applicability and limitations

    International Nuclear Information System (INIS)

    The subject of spot prices and their relationship to long-term contracting is addressed. The author is associated with Nuexco, which originally was called the Nuclear Exchange Corporation. They use the term Exchange Value which originated in the idea that Nuexco operated an exchange 'bank' - those with too much uranium could 'bank it', those with short-term needs could borrow from the 'bank'. If the borrower repaid slightly more or less the difference was settled using the 'exchange value'. This became used for longer-term transactions and now settling the monthly value is an important part of Nuexco's activities. The exact nature of the Exchange Value is defined. Now more and more buyers are insisting on spot market related pricing even where this is not meaningfully related to uranium production costs. (U.K.)

  18. TIME RESOLVED X-RAY SPOT DIAGNOSTIC

    International Nuclear Information System (INIS)

    A diagnostic was developed for the determination of temporal history of an X-ray spot. A pair of thin (0.5 mm) slits image the x-ray spot to a fast scintillator which is coupled to a fast detector, thus sampling a slice of the X-Ray spot. Two other scintillator/detectors are used to determine the position of the spot and total forward dose. The slit signal is normalized to the dose and the resulting signal is analyzed to get the spot size. The position information is used to compensate for small changes due to spot motion and misalignment. The time resolution of the diagnostic is about 1 ns and measures spots from 0.5 mm to over 3 mm. The theory and equations used to calculate spot size and position are presented, as well as data. The calculations assume a symmetric, Gaussian spot. The spot data is generated by the ETA II accelerator, a 2kA, 5.5 MeV, 60 ns electron beam focused on a Tantalum target. The spot generated is typically about 1 mm FWHM. Comparisons are made to an X-ray pinhole camera which images the X-Ray spot (in 2D) at four time slices

  19. WSSV ie1 promoter is more efficient than CMV promoter to express H5 hemagglutinin from influenza virus in baculovirus as a chicken vaccine

    Directory of Open Access Journals (Sweden)

    Yu Li

    2008-12-01

    Full Text Available Abstract Background The worldwide outbreak of influenza A (H5N1 viruses among poultry species and humans highlighted the need to develop efficacious and safe vaccines based on efficient and scaleable production. Results White spot syndrome virus (WSSV immediate-early promoter one (ie1 was shown to be a stronger promoter for gene expression in insect cells compared with Cytomegalovirus immediate-early (CMV promoter in luciferase assays. In an attempt to improve expression efficiency, a recombinant baculovirus was constructed expressing hemagglutinin (HA of H5N1 influenza virus under the control of WSSV ie1 promoter. HA expression in SF9 cells increased significantly with baculovirus under WSSV ie1 promoter, compared with CMV promoter based on HA contents and hemagglutination activity. Further, immunization with baculovirus under WSSV ie1 promoter in chickens elicited higher level anti-HA antibodies compared to CMV promoter, as indicated in hemagglutination inhibition, virus neutralization and enzyme-linked immunosorbent assays. By immunohistochemistry, strong HA antigen expression was observed in different chicken organs with vaccination of WSSV ie1 promoter controlled baculovirus, confirming higher efficiency in HA expression by WSSV ie1 promoter. Conclusion The production of H5 HA by baculovirus was enhanced with WSSV ie1 promoter, especially compared with CMV promoter. This contributed to effective elicitation of HA-specific antibody in vaccinated chickens. This study provides an alternative choice for baculovirus based vaccine production.

  20. Glare Spot Phase Doppler Anemometry

    OpenAIRE

    Hespel, Camille; Ren, Kuan Fang; Gre?han, Ge?rard; Onofri, Fabrice

    2006-01-01

    The Phase Doppler anemometry has been developed to measure simultaneously the velocity and the size of droplets. The measurement of the refractive index is also necessary since it depends on the temperature and the composition of the particle and its measurement permits both to increase the quality of the diameter measurement and to obtain information on the temperature and/or the composition of the droplets. In this paper, we introduce a Glare Spot Phase Doppler Anemometry which uses two lar...

  1. Measurement of laser spot quality

    Science.gov (United States)

    Milster, T. D.; Treptau, J. P.

    1991-01-01

    Several ways of measuring spot quality are compared. We examine in detail various figures of merit such as full width at half maximum (FWHM), full width at 1/(e exp 2) maximum, Strehl ratio, and encircled energy. Our application is optical data storage, but results can be applied to other areas like space communications and high energy lasers. We found that the optimum figure of merit in many cases is Strehl ratio.

  2. Lightning Hot Spots in Canada

    Science.gov (United States)

    Developed by the Meteorological Service of Canada, this Web site is devoted to lightning phenomena. Occurring 2.7 million times a year in Canada, visitors can find maps of Hot Spots and of current Lightning Activity over Canada. Educators and students can obtain lots of materials dealing with lightning properties, occurrences, and destructive capabilities. The site provides information on lightning activity differences among various geographic regions in Canada. Families can benefit from the high-quality and thorough, Lightning Safety Tips.

  3. 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…

  4. Smith-McGuckin spot.

    OpenAIRE

    Smith, C. W.

    1985-01-01

    Direct inspection of the tympanic membrane, especially with the help of the microscope, continues to be an important part of the examination of the patient suffering from ear disease. Accurate assessment of the varied appearances can be helpful in anticipating the prognosis in SMOM and may give information about the pathology of CSOM and cholesteatoma. The Smith-McGuckin spot was first described 20 years ago (Smith 1964, R Thomas 1965, personal communication) and its significance is reviewed....

  5. The Bandana Game: Spotted Dolphins

    Science.gov (United States)

    Jonathan Bird Productions

    2007-03-01

    In the open ocean around the Bahamas, pods of wild Spotted Dolphins frolic in the sunshine. Sometimes, they get bored and approach boats. In this educational video, Jonathan joins dolphin expert Wayne Scott Smith to learn how dolphins interact with each other. Jonathan learns how to play the Bandana Game, a game of -keep away- that the dolphins invented and like to play with Scott. Please see the accompanying study guide for educational objectives and discussion points.

  6. Recombinant expression and characterization of a serine protease inhibitor (Lvserpin7) from the Pacific white shrimp, Litopenaeus vannamei.

    Science.gov (United States)

    Liu, Yongjie; Hou, Fujun; Wang, Xianzong; Liu, Xiaolin

    2015-02-01

    Serine protease inhibitors (serpins) are widely known to its inhibitory role on proteases involved in the immune responses. Herein, a novel serine protease inhibitor (Lvserpin7), encoding for 411 amino acids with calculated molecular mass of 46.29 kDa and isoelectric point of 6.98 was characterized from the Pacific white shrimp Litopenaeus vannamei. Lvserpin7 shared 92.9% identities to Penaeus monodon serpin7. Among the tested tissues, Lvserpin7 was mainly expressed in hemocytes and gill. The expression profiles analysis indicated that Lvserpin7 was significantly up-regulated in the early stage upon Vibrio anguillarum, Micrococcus lysodeikticus or White Spot Syndrome Virus (WSSV) infection. Fusion protein expression was induced by IPTG, and the purified recombinant Lvserpin7 protein (rLvserpin7) binds to both the Gram-positive and Gram-negative bacteria. Also rLvserpin7 exhibited inhibitory activity against the proteases secreted by Bacillus subtilis. Moreover, rLvserpin7 showed inhibition role on prophenoloxidase activation. To recap, we proposed that Lvserpin7 was implicated in the shrimp immunity via the inhibition of bacterial proteases and proteases involved in prophenoloxidase system. PMID:25462553

  7. Comparison of immune response of Pacific white shrimp, Litopenaeus vannamei, after multiple and single infections with WSSV and Vibrio anguillarum.

    Science.gov (United States)

    Qiao, Guo; Xu, De-Hai; Wang, Zishen; Jang, In-Kwon; Qi, Zhitao; Zhang, Mingming; Kim, Su-Kyoung

    2015-05-01

    Our previous study demonstrated that Pacific white shrimp (Litopenaeus vannamei) infected by multiple pathogens showed higher mortality and death occurred more quickly than those infected by a single pathogen (Jang et al., 2014). For better understanding the defense mechanism against white spot syndrome virus (WSSV) and Vibrio anguillarum, immune responses of shrimp were evaluated in this study. The mRNA expression levels of five immune-related genes were analyzed by quantitative reverse real-time PCR, which included proPO-activating enzyme 1 (PPAE1), PPAE2, proPO activating factor (PPAF), masquerade-like serine proteinase (Mas) and ras-related nuclear gene (Ran). Results demonstrated that the transcription was suppressed more intensively in the multiple infection group than those in single infection groups. The transcriptional suppression was directly related to the higher mortality. The hypoimmunity could benefit pathogen invasion, replication and release of toxin in vivo. Results in this study will help to understand immune defense mechanism after shrimp were infected by multiple pathogens in aquaculture. PMID:25700782

  8. Identification, characterization and functional analysis of a serine protease inhibitor (Lvserpin) from the Pacific white shrimp, Litopenaeus vannamei.

    Science.gov (United States)

    Liu, Yongjie; Hou, Fujun; He, Shulin; Qian, Zhaoying; Wang, Xianzong; Mao, Aitao; Sun, Chengbo; Liu, Xiaolin

    2014-03-01

    As important arthropod immune responses, prophenoloxidase (proPO) activation and Toll pathway initiation are mediated by serine proteinase cascades and regulated by serpins. Herein, a serine protease inhibitor (Lvserpin), encoding for 415 amino acids with calculated molecular weight of 46,639 Da and isoelectric point of 7.03 was characterized from the Pacific white shrimp Litopenaeus vannamei. Multiple sequence alignment revealed that Lvserpin shared the highest similarity with Penaeus monodon serpin6 (87%). Quantitative real-time PCR (qRT-PCR) results showed that the transcripts of Lvserpin were detected in all the examined tissues and most highly expressed in gill. The expression profiles of Lvserpin were greatly fluctuated upon infection of Vibrio anguillarum, Micrococcus lysoleikticus or White Spot Syndrome Virus (WSSV). Double stranded RNA-mediated suppression of Lvserpin resulted in a significant increase in the transcripts of two clip-domain serine proteinases (PPAE and PPAF), prophenoloxidase (proPO), anti-lipopolysaccharide factor (ALF), Crustin and penaeidin3 (Pens3) and also increased the high cumulative mortality post V. anguillarum injection. Besides, the recombinant Lvserpin protein (rLvserpin) was purified and exhibited inhibitory activity against trypsin. Also the rLvserpin showed inhibition on prophenoloxidase activation and bacterial growth. Hence, we proposed that the Lvserpin played important role in the shrimp innate immunity. PMID:24211341

  9. [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. PMID:25876358

  10. Presence and Distribution of Tobacco Viruses in Montenegro

    Directory of Open Access Journals (Sweden)

    Jelena Zindovi?

    2007-01-01

    Full Text Available Seven important tobacco viruses were investigated in Montenegro in 2005: Tobacco Mosaic Virus (TMV, Tomato Spotted Wilt Virus (TSWV, Cucumber Mosaic Virus (CMV, Potato Virus Y (PVY, Alfalfa Mosaic Virus (AMV, Tobacco Ring Spot Virus (TRSV and Potato Virus X(PVX. This investigation included sample collection from four tobacco growing regions in Montenegro and their serological testing by DAS-ELISA test. Presence of different strains of PVY was investigated as well using DAS ELISA test with specific monoclonal antibodies.Serological results proved the presence of four tobacco viruses (TMV, CMV, PVY and AMV, while TSWV, TRSV and PVX were not found in the tested samples of tobacco crops in Montenegro. The results also showed that TMV and CMV were the most frequent (44.6% and 41.5% of tested samples, respectively followed by PVY (15.4% and the least frequent AMV (3.1%. Most samples were infected with one of the examined viruses. In the PVY population found in Montenegro, its necrotic strain (PVYN was absolutely predominant.The results indicated the significance of TMV and CMV concerning tobacco viral infections in Montenegro, as well as a necessity of their detailed characterization at biological and molecular level.

  11. CHLORELLA VIRUSES

    OpenAIRE

    Yamada, Takashi; ??, ?.; ????, ???; Onimatsu, Hideki; Etten, James L.

    2006-01-01

    Chlorella viruses or chloroviruses are large, icosahedral, plaque?forming, double?stranded?DNA—containing viruses that replicate in certain strains of the unicellular green alga Chlorella. DNA sequence analysis of the 330?kbp genome of Paramecium bursaria chlorella virus 1 (PBCV?1), the prototype of this virus family (Phycodnaviridae), predict ?366 protein?encoding genes and 11 tRNA genes. The predicted gene products of ?50% of these genes resemble proteins of known function...

  12. Integrating Plant Essential Oils and Kaolin for the Sustainable Management of Thrips and Tomato Spotted Wilt on Tomato

    Science.gov (United States)

    Thrips-vectored Tomato spotted wilt virus is one of the most devastating pest complexes affecting tomato in the southern USA and elsewhere. Field trials were conducted over two years to determine the effects of volatile plant essential oils and kaolin based particle films on the incidence of Tomato...

  13. Energy is not Coffee. An assessment of blind spots on energy spot-markets

    International Nuclear Information System (INIS)

    This study was to be the first in a series of studies on the title subject. It specifically focuses on the differences and similarities with a number of other spot-markets and aims to frame the energy spot markets and their potential development into a broader perspective. Main conclusion is that energy spot-markets differ from several other physical and non-physical spot-markets in many ways. This implies that 'perfect' energy spot-markets may inherently be (much) less perfect than other spot-markets that have approximated the stage of theoretical perfection

  14. Energy is not Coffee. An assessment of blind spots on energy spot-markets

    Energy Technology Data Exchange (ETDEWEB)

    Jepma, C.J.; Spijker, E.; Van der Gaast, W.; De Jong, F. [Foundation Joint Implementation Network JIN, Paterswolde (Netherlands); Overmars, P. (ed.) [Energy Delta Institute EDI, Groningen (Netherlands)

    2006-01-15

    This study was to be the first in a series of studies on the title subject. It specifically focuses on the differences and similarities with a number of other spot-markets and aims to frame the energy spot markets and their potential development into a broader perspective. Main conclusion is that energy spot-markets differ from several other physical and non-physical spot-markets in many ways. This implies that 'perfect' energy spot-markets may inherently be (much) less perfect than other spot-markets that have approximated the stage of theoretical perfection.

  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. ECHO virus

    Science.gov (United States)

    Enteric cytopathic human orphan (ECHO) viruses are a group of viruses that lead to gastrointestinal infection and skin rashes. ... Echovirus is one of several families of viruses that affect the ... are common. In the United States, they are most common in ...

  17. Spot it(R) Solitaire

    OpenAIRE

    Dietz, Donna A.

    2013-01-01

    The game of Spot it(R) is based on an order 7 finite projective plane. This article presents a solitaire challenge: extract an order 7 affine plane and arrange those 49 cards into a square such that the symmetries of the affine and projective planes are obvious. The objective is not to simply create such a deck already in this solved position. Rather, it is to solve the inverse problem of arranging the cards of such a deck which has already been created shuffled.

  18. Exploitation of the SPOT system

    Science.gov (United States)

    Calhes, G.; Trempat, Y.

    After briefly describing the organizational structure, the paper describes the different components of the SPOT ground system, which include the Mission and Operations Control Center in charge of controlling all the exploitation operations, the Space Imagery Receiving Station which collects the raw data from the satellite telemetry, and the Space Imagery Rectification Center, which performs the preprocessing of these data into the standard products to be offered to the customers. Indications are given regarding the capacity of the system in terms of the number of images which it can acquire and process per year and per receiving station, as a function of their latitude.

  19. Chemical composition on cacao leaves infected by viruses

    International Nuclear Information System (INIS)

    Chemical analysis on cacao leaves that have chlorosis spots caused by cocoa swollen shoot viruses were carried out. It can be shown that leaves with chlorosis spots contain less chlorophyl and lipides than those without, but both do not show any significant difference in the concentration of water, glucose, saccharides, amino acid and proteins. It can be concluded that transport systems in the infected leaves are good so that the water and saccharides distribution in them are not disturbed. (author tr.)

  20. Inhibitory effect of Newcastle disease virus on hepatic fibrosis induced by CCl4

    Directory of Open Access Journals (Sweden)

    Ya-lin LI

    2013-11-01

    Full Text Available Objective?To explore the inhibitory effect of Newcastle diseases virus (NDV on CCl4-induced liver fibrosis in mice. Methods?Liver fibrosis model was reproduced in 30 Kunming mice by intraperitoneal injection of CCl4/peanut oil solution for 2 times a week, and the total treatment lasted for 8 weeks. Three days after last injection, NDV was injected through tail vein for 1 or 3 times (24h intervals. Twenty-four hours after NDV infusion, mice were sacrificed and the livers were removed for gross morphology observation. The liver tissue sections were stained by HE and Sirius red dyeing. ?-smooth muscle actin (?-SMA expression was detected by Western blotting. Results?After CCl4 induction for 8 weeks, obvious fibrosis symptoms appeared in the liver of model mice, and the surface of liver tissue became hard with rough, with white patches on it. HE staining showed that there was loosening of tissue and enlarged perisinusoidal spaces in liver with fibrosis. Sirius red dyeing displayed abnormal collagen deposition in the fibrotic liver tissues. After NDV injection for 3 times, white spots on the surface of mouse liver were significantly reduced, and collagen deposition was lowered. Western blotting showed that ?-SMA levels decreased with increasing frequency of NDV injection. Conclusion?NDV may effectively suppress the development of CCl4-induced liver fibrosis in mice. DOI: 10.11855/j.issn.0577-7402.2013.11.003

  1. Activation of PmRelish from Penaeus monodon by yellow head virus.

    Science.gov (United States)

    Visetnan, Suwattana; Supungul, Premruethai; Hirono, Ikuo; Tassanakajon, Anchalee; Rimphanitchayakit, Vichien

    2015-02-01

    Humoral innate immune response against pathogenic infection is partly responsible by the Imd pathway in which a transcription factor Relish relays the infection signals to the nuclei for the expression of antimicrobial proteins. A PmRelish gene which encoded a protein of 1195 amino acids was cloned. The PmRelish was constitutively expressed in all tissues tested and mostly up-regulated upon YHV infection. In hemocytes, the PmRelish expression was up-regulated upon Vibrio harveyi, yellow head virus (YHV) and white spot syndrome virus (WSSV) challenges. Using dsRNA silencing of PmRelish gene, it was shown that the expression of penaeidin5 but not anti-lipopolysaccharide factor ALFPm3, crustinPm1 and penaeidin3 was under the regulation of Imd pathway. Under PmRelish silencing, the shrimp were more susceptible to infection by YHV with the 50% survival rate reduced from about 72 h to 42 h. The PmRelish was detected in the cytoplasm of all the hemocytes from both uninfected and YHV-infected shrimp. The accumulation of activated PmRelish in the nuclei was not clearly observed but the activated PmRelish was detected in the YHV-infected hemocytes by Western blot analysis. Thus, the PmRelish and, hence, the Imd pathway respond to the YHV infection. PMID:25463289

  2. Transcriptome analysis of orange-spotted grouper (Epinephelus coioides) spleen in response to Singapore grouper iridovirus

    OpenAIRE

    Huang Youhua; Huang Xiaohong; Yan Yang; Cai Jia; Ouyang Zhengliang; Cui Huachun; Wang Peiran; Qin Qiwei

    2011-01-01

    Abstract Background Orange-spotted grouper (Epinephelus coioides) is an economically important marine fish cultured in China and Southeast Asian countries. The emergence of infectious viral diseases, including iridovirus and betanodavirus, have severely affected food products based on this species, causing heavy economic losses. Limited available information on the genomics of E. coioides has hampered the understanding of the molecular mechanisms that underlie host-virus interactions. In this...

  3. Computational Prediction of Hot Spot Residues

    OpenAIRE

    Morrow, John Kenneth; Zhang, Shuxing

    2012-01-01

    Most biological processes involve multiple proteins interacting with each other. It has been recently discovered that certain residues in these protein-protein interactions, which are called hot spots, contribute more significantly to binding affinity than others. Hot spot residues have unique and diverse energetic properties that make them challenging yet important targets in the modulation of protein-protein complexes. Design of therapeutic agents that interact with hot spot residues has pr...

  4. Spot market power and future market trading

    OpenAIRE

    Muermann, Alexander; Shore, Stephen H.

    2005-01-01

    When a spot market monopolist participates in the futures market, he has an incentive to adjust spot prices to make his futures market position more pro…table. Rational futures market makers take this into account when they set prices. Spot market power thus creates a moral hazard problem which parallels the adverse selection problem in models with inside information. This moral hazard not only reduces the optimal amount of hedging for those with and without market power, but also makes com...

  5. Turbulent spots in a Stokes boundary layer

    International Nuclear Information System (INIS)

    The turbulent spots which form in a boundary layer generated by the harmonic oscillations of an incompressible fluid are investigated by numerical means. In order to allow the formation of turbulent spots, the dimensions of the computational box have been increased with respect to previous numerical investigations (Costamagna et al. (2003)). The boundaries of the spots are identified and the speeds of the head, tail, leftmost and rightmost points, are computed. The computed speeds well compare with those measured in steady boundary layers.

  6. Oil futures and spot markets

    International Nuclear Information System (INIS)

    In the last decade, the oil futures market has risen to prominence and has become a major factor in influencing oil market psychology and the crude oil market. On a normal day, over 92 thousand contracts, the equivalent of 92 million barrels per day, change hands on the New York Mercantile Exchange, NYMEX. This market has provided a vehicle for hedging against risk. At the same time, it has also created opportunities for speculation. Those who previously were unable to participate in oil market transactions can now become involved through the futures market. The large number of participants in the future market and the availability of information has made this market more efficient and transparent, relative to the crude oil market. While there has been considerable in-depth analysis of other future markets, relatively little theoretical attention has focused on that of oil. This paper looks at the following issues. First, what is the relationship between futures and spot oil prices? And secondly, are futures prices a good predictor of spot crude prices in the future? (author)

  7. Poisson’s spot with molecules

    OpenAIRE

    Reisinger, Thomas; Patel, Amil A.; Reingruber, Herbert; Fladischer, Katrin; Ernst, Wolfgang E.; Bracco, Gianangelo; Smith, Henry I.; Holst, Bodil

    2009-01-01

    In the Poisson-spot experiment, waves emanating from a source are blocked by a circular obstacle. Due to their positive on-axis interference an image of the source the Poisson spot is observed within the geometrical shadow of the obstacle. In this paper we report the observation of Poisson’s spot using a beam of neutral deuterium molecules. The wavelength independence and the weak constraints on angular alignment and position of the circular obstacle make Poisson’s spot a pr...

  8. Gene expression analysis identifies new candidate genes associated with the development of black skin spots in Corriedale sheep.

    Science.gov (United States)

    Peñagaricano, Francisco; Zorrilla, Pilar; Naya, Hugo; Robello, Carlos; Urioste, Jorge I

    2012-02-01

    The white coat colour of sheep is an important economic trait. For unknown reasons, some animals are born with, and others develop with time, black skin spots that can also produce pigmented fibres. The presence of pigmented fibres in the white wool significantly decreases the fibre quality. The aim of this work was to study gene expression in black spots (with and without pigmented fibres) and white skin by microarray techniques, in order to identify the possible genes involved in the development of this trait. Five unrelated Corriedale sheep were used and, for each animal, the three possible comparisons (three different hybridisations) between the three samples of interest were performed. Differential gene expression patterns were analysed using different t-test approaches. Most of the major genes with well-known roles in skin pigmentation, e.g. ASIP, MC1R and C-KIT, showed no significant difference in the gene expression between white skin and black spots. On the other hand, many of the differentially expressed genes (raw P-value?spots. The gene expression of C-FOS and KLF4, transcription factors involved in the cellular response to external factors such as ultraviolet light, was validated by quantitative polymerase chain reaction (PCR). This exploratory study provides a list of candidate genes that could be associated with the development of black skin spots that should be studied in more detail. Characterisation of these genes will enable us to discern the molecular mechanisms involved in the development of this feature and, hence, increase our understanding of melanocyte biology and skin pigmentation. In sheep, understanding this phenomenon is a first step towards developing molecular tools to assist in the selection against the presence of pigmented fibres in white wool. PMID:21952730

  9. Dried Blood Spots - Preparing and Processing for Use in Immunoassays and in Molecular Techniques

    Science.gov (United States)

    Grüner, Nico; Stambouli, Oumaima; Ross, R. Stefan

    2015-01-01

    The idea of collecting blood on a paper card and subsequently using the dried blood spots (DBS) for diagnostic purposes originated a century ago. Since then, DBS testing for decades has remained predominantly focused on the diagnosis of infectious diseases especially in resource-limited settings or the systematic screening of newborns for inherited metabolic disorders and only recently have a variety of new and innovative DBS applications begun to emerge. For many years, pre-analytical variables were only inappropriately considered in the field of DBS testing and even today, with the exception of newborn screening, the entire pre-analytical phase, which comprises the preparation and processing of DBS for their final analysis has not been standardized. Given this background, a comprehensive step-by-step protocol, which covers al the essential phases, is proposed, i.e., collection of blood; preparation of blood spots; drying of blood spots; storage and transportation of DBS; elution of DBS, and finally analyses of DBS eluates. The effectiveness of this protocol was first evaluated with 1,762 coupled serum/DBS pairs for detecting markers of hepatitis B virus, hepatitis C virus, and human immunodeficiency virus infections on an automated analytical platform. In a second step, the protocol was utilized during a pilot study, which was conducted on active drug users in the German cities of Berlin and Essen. PMID:25867233

  10. Analysis on the expression and function of syndecan in the Pacific white shrimp Litopenaeus vannamei.

    Science.gov (United States)

    Yang, Hui; Li, Shihao; Li, Fuhua; Wen, Rong; Xiang, Jianhai

    2015-08-01

    Syndecan is considered to be a multifunctional protein which functions as a cell surface receptor involved in cell adhesion, migration, cytoskeleton organization and differentiation. Previous bioinformatic analysis has revealed that syndecan in shrimp might interact with white spot syndrome virus (WSSV). In the present study, we experimentally studied the function of syndecan in shrimp immunity. The syndecan from Litopenaeus vannamei (LvSDC) was cloned and analyzed. The full-length cDNA of LvSDC was 1005?bp, consisting of 59?bp 5'-UTR, 253?bp 3'-UTR, and 693?bp open reading frame encoding 230 amino acids. LvSDC consisted of an extracellular domain (ED), a transmembrane domain (TM) and a cytoplasmic domain (CD). TM and CD shared high similarities with those of syndecan proteins from other species. LvSDC was ubiquitously expressed in all tested tissues, with the highest level in Oka. After WSSV challenge, the transcription level of LvSDC in Oka was apparently up-regulated. Recombinant LvSDC protein and its rabbit polyclonal antibody were prepared for detecting the location of LvSDC in hemocytes using immunocytochemistry approach. Data showed that LvSDC mainly located at the cell membrane and the cytoplasm of hemocytes. After silencing of LvSDC with siRNA, the WSSV copy numbers and mortality of shrimp after WSSV infection were both significantly decreased. These data provide useful information for understanding the immune mechanism of shrimp to WSSV infection. PMID:25847874

  11. RealSpot: software validating results from DNA microarray data analysis with spot images.

    Science.gov (United States)

    Chen, Zhongming; Liu, Lin

    2005-04-14

    The spot images from DNA microarray highly affect the discovery of biological knowledge from gene expression data. However, results from quality analysis, normalization, differential expression, and cluster analysis are rarely validated with spot images in current data analysis methods or software packages. We designed RealSpot, a software package, to validate the results by directly associating spot quality and data with spot images in a spreadsheet table. RealSpot splits hybridization images into individual spots stored in a spreadsheet table. It subsequently associates microarray data with spot images and performs data validation through the standard table operation such as sorting, searching, and editing. RealSpot has several built-in functions to facilitate data validation, including spot quality analysis, data organization, one-way ANOVA, gene ontology association, verification, import, and export. We used RealSpot to evaluate 77 slides (30,000 features each) from real hybridization experiments and to validate results from each step of data analysis. It took approximately 10 min to validate results of spot quality after initial evaluation and correct approximately 0.3% of falsely assigned qualities of 10,000 spots. We validated 1,641 of 2,110 differentially expressed genes identified by SAM analysis in approximately 1/2 h by comparing each gene with its respective spot image. Furthermore, we found that 6 of 48 genes in one cluster from k-mean clustering method showed inconsistent trends of spot images. RealSpot is efficient for validating microarray results and thus helpful for improving the reliability of the whole microarray experiment for experimentalists. PMID:15687483

  12. Hot White Dwarfs

    OpenAIRE

    Sion, Edward M.

    2011-01-01

    The article covers the physical properties and evolution of single white dwarfs ranging in temperature from 20,000K to 200,000 and higher, the hottest know electron-degenerate stars. After discussing the classification of their spectra, the author reviews the known properties, parameters, evolutionary state, as well as persisting and new puzzles regarding all spectroscopic subclasses of hot white dwarfs: the hot DA white dwarfs, the DAO white dwarfs, the PG1159 degenerates, ...

  13. [Semilethal spotted mutants of Chlorella].

    Science.gov (United States)

    Shabanova, E A

    1975-01-01

    Mutant colonies of Chorella vulgaris Beijer, having the region of colourless cells either in the center or in the periphery of a colony, are found. Spontaneous mutation rate varied within 1-40-10(5). The size of a mutant colony comprised about 1/2-1/3 of the diameter of a middle initial strain colony. Colonies with a mutant sector were also observed besides completely mutant colonies. Ways of the formation of mutant phenotypes are studied. It is found that from the initial spotted green colony those of the type "death in the periphery" and, through a number of intermediate stages, "death in the center" are formed. The colony development can stop at some successive stage or to result in the death of all the cells. The period of forming the mutant phenotype is found to depend on the colony growth rate. PMID:1228059

  14. Hot spots of mutualistic networks.

    Science.gov (United States)

    Gilarranz, Luis J; Sabatino, Malena; Aizen, Marcelo A; Bascompte, Jordi

    2014-11-17

    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. PMID:25402941

  15. A spot market for electricity

    International Nuclear Information System (INIS)

    Electric utilities in many parts of the world are being transformed from regulated monopolies to competitive enterprises. They are buying power from sources they don't control, and experiencing new competitive pressures in selling. In response, utilities have begun to use such techniques as ''active demand-side management'' and ''real-time pricing'' to improve their responsiveness to customers and to create new opportunities for revenue. As an extension of the industry's evolution so far, this article describes how the utilities industry might develop a ''spot market'' for electricity, with the utility providing value in the form of a service for arbitrating requests and demands from various generators and users. The author presents a possible future organization of the electricity market, for which there is no model currently known. Most of its elements do exist in some form at one or more power companies around the world

  16. Function and regulation domains of a newly isolated putative ?-actin promoter from pacific white shrimp.

    Science.gov (United States)

    Shi, Yingli; Soderlund, Marcus; Xiang, Jianhai; Lu, Yuanan

    2015-01-01

    Current development of transgenic shrimp research has been hampered due to the lack of the suitable promoters and efficient transfection methods for crustaceans. A 1642 bp sequence, containing 5'-upstream sequence, exon 1, intron 1 and partial exon 2, which is responsible for transcriptional initiation of the newly reported shrimp ?-actin (actinT1), has been isolated from the Pacific white shrimp (Litopenaeus vannamei) and named as SbaP. To determine its function and potential application in marine biotechnology, the sequence and functional domains were examined by constitutive expression of the luciferase reporter gene. We have identified 5' regions that play a central role in the expression of the ?-actin gene. The proximal promoter (-1642/-1325) contains two highly conserved transcriptional sites, CCAAT box and CArG motif. Two negative (-1140/-924, -222/-21) and one positive (-810/-425) regulatory elements have been identified in intron1. Transient transfection assay with a construct containing proximal promoter and enhancer (SbaP?-222/+1?-1325/-924) regions of the shrimp ?-actin coupled with luciferase and EGFP (enhanced green fluorescent protein) showed that the promoter was not only functional in sf21 cells, but promoter activity was more than 8-fold higher than a viral-origin promoter (ie1, white spot syndrome virus immediate early gene promoter). Furthermore, SbaP?-222/+1?-1325/-924 drove a successful expression of luciferase injection assay in vivo injection and also showed higher promoter activity than the ie1 promoter, suggesting that the expression vectors constructed with SbaP?-222/+1?-1325/-924 have important potential in gene transfer studies for shrimp and other crustacean species. PMID:25835297

  17. Ambystoma maculatum (spotted salamander). Reproduction

    Science.gov (United States)

    Glorioso, Brad M.; Waddle, Hardin; Hefner, Jeromi

    2012-01-01

    The Spotted Salamander is a wide-ranging salamander of the eastern United States that typically breeds in winter or early spring in ephemeral pools in lowland forests. Ambystoma maculatum is known to deposit 2-4 egg masses per year, each containing 1-250 eggs. As part of ongoing research into the ecology and reproductive biology of Spotted Salamanders in the Kisatchie District of Kisatchie National Forest in Natchitoches Parish, Louisiana, USA, we have been counting the number of embryos per egg mass. We captured seven female A. maculatum in a small pool, six of which were still gravid. We took standard measurements, including SVL, and then implanted a Passive Integrated Transponder (PIT tag) into each adult female as was the protocol. About an hour after processing these animals we marked new A. maculatum egg masses found in the same small pool using PVC pin flags pushed carefully through the outer jelly. We did not have enough time to process them that evening, and it was not until a few days later that we photographed those masses. We discovered that one of the masses contained a PIT tag in the outer jelly that corresponded to one of the six gravid females that were marked that same evening. To our knowledge, this is the first report of PIT tags being the means, albeit coincidentally, by which a particular egg mass of Ambystoma maculatum has been assigned to a particular female. For our purposes, losing the PIT tag from the adult female is counter to the goals of our study of this population, and we will no longer be implanting PIT tags into gravid females.

  18. Rethinking White Supremacy

    OpenAIRE

    Gillborn, David

    2006-01-01

    Abstract The article addresses the nature of power relations that sustain and disguise white racial hegemony in contemporary ‘western’ society. Following the insights offered by critical race theory (CRT), white supremacy is conceived as a comprehensive condition whereby the interests and perceptions of white subjects are continually placed centre stage and assumed as ‘normal’. These ...

  19. Probabilistic modelling of the high-pressure arc cathode spot displacement dynamic

    CERN Document Server

    Coulombe, S

    2003-01-01

    A probabilistic modelling approach for the study of the cathode spot displacement dynamic in high-pressure arc systems is developed in an attempt to interpret the observed voltage fluctuations. The general framework of the model allows to define simple, probabilistic displacement rules, the so-called cathode spot dynamic rules, for various possible surface states (un-arced metal, arced, contaminated) and to study the resulting dynamic of the cathode spot displacements over one or several arc passages. The displacements of the type-A cathode spot (macro-spot) in a magnetically rotating arc using concentric electrodes made up of either clean or contaminated metal surfaces is considered. Experimental observations for this system revealed a 1/f sup - sup t sup i sup l sup d sup e sup 1 signature in the frequency power spectrum (FPS) of the arc voltage for anchoring arc conditions on the cathode (e.g. clean metal surface), while it shows a 'white noise' signature for conditions favouring a smooth movement (e.g. ox...

  20. Rocky Mountain spotted fever in Argentina

    Science.gov (United States)

    Cases of epidemic typhus have been documented in Argentina since 1919; however, no confirmed reports of spotted fever rickettsiosis were described in this country until 1999. We describe the first molecular confirmation of Rickettsia rickettsii, the etiologic agent of Rocky Mountain spotted fever (R...

  1. 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.

  2. Hot Spot Removal System: System description

    International Nuclear Information System (INIS)

    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

  3. Rocky Mountain Spotted Fever in Argentina

    Science.gov (United States)

    We describe the first molecular confirmation of Rickettsia rickettsii, the cause of Rocky Mountain spotted fever (RMSF), from a tick vector, Amblyomma cajennense, and from a cluster of fatal spotted fever cases in Argentina. Questing A. cajennense ticks were collected at or near sites of presumed or...

  4. Modelling of the Resistance Spot Welding Process

    OpenAIRE

    Govik, Alexander

    2009-01-01

    A literature survey on modelling of the resistance spot welding process has been carried out and some of the more interesting models on this subject have been reviewed in this work. The underlying physics has been studied and a brief explanation of Heat transfer, electrokinetics and metallurgy in a resistance spot welding context have been presented.\

  5. Spotted knapweed, Centaurea stoebe, control options

    Science.gov (United States)

    Spotted knapweed is a non native perennial forb that is spreading rapidly in the Western United States. This plant species produces a compound that retards the growth of many native plants, giving it a competitive advantage. Spotted knapweed has been identified in several areas of Alaska. A descript...

  6. An Invertebrate Warburg Effect: A Shrimp Virus Achieves Successful Replication by Altering the Host Metabolome via the PI3K-Akt-mTOR Pathway

    Science.gov (United States)

    Su, Mei-An; Huang, Yun-Tzu; Chen, I-Tung; Lee, Der-Yen; Hsieh, Yun-Chieh; Li, Chun-Yuan; Ng, Tze Hann; Liang, Suh-Yuen; Lin, Shu-Yu; Huang, Shiao-Wei; Chiang, Yi-An; Yu, Hon-Tsen; Khoo, Kay-Hooi; Chang, Geen-Dong; Lo, Chu-Fang; Wang, Han-Ching

    2014-01-01

    In this study, we used a systems biology approach to investigate changes in the proteome and metabolome of shrimp hemocytes infected by the invertebrate virus WSSV (white spot syndrome virus) at the viral genome replication stage (12 hpi) and the late stage (24 hpi). At 12 hpi, but not at 24 hpi, there was significant up-regulation of the markers of several metabolic pathways associated with the vertebrate Warburg effect (or aerobic glycolysis), including glycolysis, the pentose phosphate pathway, nucleotide biosynthesis, glutaminolysis and amino acid biosynthesis. We show that the PI3K-Akt-mTOR pathway was of central importance in triggering this WSSV-induced Warburg effect. Although dsRNA silencing of the mTORC1 activator Rheb had only a relatively minor impact on WSSV replication, in vivo chemical inhibition of Akt, mTORC1 and mTORC2 suppressed the WSSV-induced Warburg effect and reduced both WSSV gene expression and viral genome replication. When the Warburg effect was suppressed by pretreatment with the mTOR inhibitor Torin 1, even the subsequent up-regulation of the TCA cycle was insufficient to satisfy the virus's requirements for energy and macromolecular precursors. The WSSV-induced Warburg effect therefore appears to be essential for successful viral replication. PMID:24945378

  7. Susceptibility to an inoculum of infectious hypodermal and haematopoietic necrosis virus (IHHNV) in three batches of whiteleg shrimp Litopenaeus vannamei (Boone, 1931)

    Science.gov (United States)

    Escobedo-Bonilla, César Marcial; Rangel, José Luis Ibarra

    2014-01-01

    Abstract The present study evaluated the susceptibility of three different batches of whiteleg shrimp Litopenaeus vannamei from Mexico to an inoculum of infectious hypodermal and haematopoietic necrosis virus (IHHNV). Each of the three shrimp batches came from a different hatchery. Because of their origin, it was possible that the genetic makeup of these batches was different among each other. The three batches tested showed differences in IHHNV susceptibility. Here, susceptibility is defined as the capacity of the host to become infected, and it can be measured by the infectivity titer. Susceptibility to IHHNV was observed in decreasing order in shrimp from batch 1 (hatchery from El Rosario, Sinaloa), batch 3 (hatchery from Nayarit) and batch 2 (hatchery from El Walamo, Sinaloa), respectively. The largest susceptibility difference between batches was 5012 times, and that between early and late juveniles from the same batch was 25 times. These results indicate that within a species, susceptibility to a pathogen such as IHHNV can have large differences. Susceptibility to pathogens is an important trait to consider before performing studies on pathogenesis. It may influence virological parameters such as speed of replication, pathogenicity and virus titer. In order to evaluate the potential use of IHHNV as a natural control agent against white spot syndrome virus (WSSV), it is necessary to know host susceptibility and the kinetics of IHHNV infection. These features can help to determine the conditions in which IHHNV could be used as antagonist in a WSSV infection. PMID:25561847

  8. A mammalian spot test: induction of genetic alterations in pigment cells or mouse embryos with X-rays and chemical mutagens

    International Nuclear Information System (INIS)

    Embryos heterozygous for five recessive coat-color genes from the cross C57 BL/6 J Han x T-stock were X-irradiated with 100 r or treated in utero with 50 mg/kg methyl methanesulfonate (MMS) and ethyl methanesulfonate (EMS), respectively. Controls consisted of irradiated embryos of C57 BL x C57 BL matings homozygous wild-type for the genes under study, and non-treated offspring of both types of mating. The colors of the spots observed in the adult fur were either due to expression of the recessive coat genes or were white. 1) Irradiated and mutagen-treated offspring of C57 BL x T-stock matings had almost exclusively nonwhite spots, distributed randomly over the mouse surface. 2) Irradiated offspring of C57 BL x C57 BL matings had only white spots which were always midventral. 3) In non-treated offspring of both types of mating no spot could be observed. It is discussed that the white midventral spots are preferentially the result of pigment cell killing, while the nonwhite spots are preferentially the result of gene mutations or recombinational processes like mitotic crossing over and mitotic gene conversion. (orig./BSC)

  9. Jupiter's New Red Spot - An Indication of Climate Change?

    Science.gov (United States)

    Marcus, Philip; Jiang, Chung-Hsiang; Asay-Davis, Xylar

    2007-11-01

    Jupiter now has two red spots. The new Red Oval, is similar to the Great Red Spot and is Jupiter's second largest storm. The Oval was originally white, but turned red in 2005. This color change was the first sign of the current jovian ``upheaval'' at the latitudes north of the Oval. Using a new method, Data Assimilated Correlation Image Velocimetry, we derive the Oval's velocities with unprecedented accuracy from spacecraft images. These data show that the velocities of the Oval did not between 2000 and 2005 and therefore cannot account for its color change. Although the dynamics of upheavals and other jovian climate cycles are not well understood, in 2001 we predicted that a large-scale warming at, and north of, the Oval, was about to begin, with effects becoming visible in 2006. Currently, no instrument can directly confirm whether the average jovian temperatures changed, and therefore we do not know whether a warming was responsible for the current upheaval or for the Oval's red color. However, we show that the Oval's color change is consistent with the location, magnitude, and timescale of the predicted temperature change. We show also that other proposed explanations for the color change are not plausible.

  10. Element spots in HgMn stars

    CERN Document Server

    Korhonen, Heidi

    2013-01-01

    A fraction of late B-type stars, the so-called HgMn stars, exhibit enhanced absorption lines of certain chemical elements, notably Hg and Mn, combined with an underabundance of He. For about a decade now the elements with anomalously high abundances in HgMn stars are known to be distributed inhomogeneously over the stellar surface. Temporal evolution of these elemental spots have been reported in a few HgMn stars, first secular evolution of the mercury spots in alpha And, and recently also a fast evolution of yttrium and strontium spots in HD 11753. The fast evolution of spots in HD 11753 is combined with a slower change in the overall abundance of the affected elements. In this paper I review what is known of elemental spots in HgMn stars and their secular and fast temporal evolution.

  11. Ultrasonic hammer produces hot spots in solids

    Science.gov (United States)

    You, Sizhu; Chen, Ming-Wei; Dlott, Dana D.; Suslick, Kenneth S.

    2015-04-01

    Mechanical action can produce dramatic physical and mechanochemical effects when the energy is spatially or temporally concentrated. An important example of such phenomena in solids is the mechanical initiation of explosions, which has long been speculated to result from ‘hot spot’ generation at localized microstructures in the energetic material. Direct experimental evidence of such hot spots, however, is exceptionally limited; mechanisms for their generation are poorly understood and methods to control their locations remain elusive. Here we report the generation of intense, localized microscale hot spots in solid composites during mild ultrasonic irradiation, directly visualized by a thermal imaging microscope. These ultrasonic hot spots, with heating rates reaching ~22,000?K?s?1, nucleate exclusively at interfacial delamination sites in composite solids. Introducing specific delamination sites by surface modification of embedded components provides precise and reliable control of hot spot locations and permits microcontrol of the initiation of reactions in energetic materials including fuel/oxidizer explosives.

  12. Análise ultra-estrutural de folhas de três espécies de Solanaceae após inoculação com o vírus da necrose branca do tomateiro (VNBT - Tymovirus) / Ultrastructural analysis of leaves from three species of Solanaceae after inoculation with the tomato white necrosis virus (TWNV - Tymovirus)

    Scientific Electronic Library Online (English)

    FRANCISCO ANDRÉ OSSAMU, TANAKA; SÍLVIA RODRIGUES, MACHADO; MARIA MÉRCIA, BARRADAS.

    2000-06-01

    Full Text Available Foi estudada a ultra-estrutura foliar das solanáceas Nicotiana glutinosa L., Lycopersicon pimpinellifolium L. e Physalis angulata L. inoculadas com o vírus da necrose branca do tomateiro (VNBT - Tymovirus). As plantas mantidas em casa-de-vegetação com temperatura constante de 25 °C foram inoculadas [...] quando apresentavam três a quatro folhas totalmente expandidas. Quinze dias após a inoculação, foram coletadas amostras do terço médio do limbo da 3ª ou da 4ª folha a partir do ápice. As amostras foram preparadas para análise em microscopia eletrônica de transmissão segundo técnicas convencionais. A análise ultra-estrutural das células do clorênquima revelou principalmente vesiculação e vacuolação dos cloroplastos e de mitocôndrias, além da ocorrência de corpos multivesiculares e ligeira dilatação dos plasmodesmos em N. glutinosa e L. pimpinellifolium. Nestas duas espécies foram observadas "virus-like-particles" no citoplasma e nos vacúolos. Plantas de P. angulata não mostraram alterações de ultra-estrutura. As observações macroscópicas, os testes de retroinoculação e as análises ultra-estruturais revelaram sintomas locais e sistêmicos em N. glutinosa, latência em L. pimpinellifolium e imunidade em P. angulata. Abstract in english The leaf ultrastructure of Nicotiana glutinosa L., Lycopersicon pimpinellifolium L., and Physalis angulata L. inoculated with the tomato white necrosis virus (TWNV - Tymovirus) was analysed. Plants were kept in a greenhouse at 25 °C and inoculated with the virus when tree to four leaves were totally [...] developed. Fifteen days after inoculation, samples were collected from the median third of the 3rd or 4th leaf from the apex. The samples were prepared for analysis using conventional transmission electron microscope techniques. The ultrastructural analysis of the chlorenquima cells showed vesiculation and vacuolation of the chloroplasts and mitochondria, presence of multivesicular bodies, and a slight dilation of the plasmodesmata in N. glutinosa and L. pimpinellifolium. In these two species, virus-like particles were seen in the cytoplasm and vacuoles. P. angulata plants did not show subcellular alterations. Macroscopic observations, retroinoculation tests, and ultrastructural analysis showed the presence of local and systemic symptoms in N. glutinosa, latency in L. pimpinellifolium, and immunity in P. angulata.

  13. Live Virus Smallpox Vaccine

    Science.gov (United States)

    ... Index SMALLPOX FACT SHEET The Live Virus Smallpox Vaccine The vaccinia virus is the "live virus" used ... cannot cause smallpox. What is a "live virus" vaccine? A "live virus" vaccine is a vaccine that ...

  14. Obesity Virus

    Science.gov (United States)

    Science Update (AAAS; )

    2007-06-12

    Obesity has many causes, but there is growing evidence that common viruses may contribute to the condition in some people. Recently, Nikhil Dhurandhar and his colleagues at the Pennington Biomedical Research Center infected human stem cells with Ad-36, a common virus known to be associated with obesity in humans. They found that the cells they exposed to the virus accumulated a much higher amount of fat than uninfected cells.

  15. Virus Diseases of Cucurbits in Gaziantep-Turkey

    OpenAIRE

    Mehmet Ozaslan; Turkan Aytekin; Berna Bas; Halil Kilic, I.; Didem Afacan, I.; Dag, D. S.

    2006-01-01

    Cucurbit growing is affected negatively due to diseases caused by cucurbit viruses. In order to prevent this damage cucurbit viruses were identified by serologically. Due to this study, it is usually difficult to give definitive diagnosis based on symptoms but occasionally symptoms; are curling, wrinkling, spot mosaics, yellowing, shape deformation on leaves, smaller leaves than normal, buff-colored mosaics, observed on younger leaves of cucurbits and stunting , distortion and fruit deformati...

  16. Detection and Identification of the First Viruses in Chia (Salvia hispanica

    Directory of Open Access Journals (Sweden)

    Marcos G. Celli

    2014-09-01

    Full Text Available Chia (Salvia hispanica, an herbaceous plant native to Latin America, has become important in the last 20 years due to its beneficial effects on health. Here, we present the first record and identification of two viruses in chia plants. The comparison of the complete nucleotide sequences showed the presence of two viral species with the typical genome organization of bipartite New World begomovirus, identified as Sida mosaic Bolivia virus 2 and Tomato yellow spot virus, according to the ICTV taxonomic criteria for begomovirus classification. DNA-A from Sida mosaic Bolivia virus 2 exhibited 96.1% nucleotide identity with a Bolivian isolate of Sida micrantha, and Tomato yellow spot virus showed 95.3% nucleotide identity with an Argentine bean isolate. This is the first report of begomoviruses infecting chia as well as of the occurrence of Sida mosaic Bolivia virus 2 in Argentina.

  17. Superluminal Spot Pair Events in Astronomical Settings: Sweeping Beams

    CERN Document Server

    Nemiroff, Robert J

    2014-01-01

    Sweeping beams of light can cast spots moving with superluminal speeds across scattering surfaces. Such faster-than-light speeds are well-known phenomena that do not violate special relativity. It is shown here that under certain circumstances, superluminal spot pair creation and annihilation events can occur that provide unique information to observers. These spot pair events are {\\it not} particle pair events -- they are the sudden creation or annihilation of a pair of relatively illuminated spots on a scattering surface. Real spot pair illumination events occur unambiguously on the scattering surface when spot speeds diverge, while virtual spot pair events are observer dependent and perceived only when real spot radial speeds cross the speed of light. Specifically, a virtual spot pair creation event will be observed when a real spot's speed toward the observer drops below $c$, while a virtual spot pair annihilation event will be observed when a real spot's radial speed away from the observer rises above $c...

  18. Full local elastic strain tensor from Laue micro-diffraction: simultaneous Laue pattern and spot energy measurement

    International Nuclear Information System (INIS)

    In sample-scanning Laue micro-diffraction, the local crystal orientation and local deviatoric strain tensor are obtained by illuminating the polycrystalline sample with a broadband 'white' (5-30 keV) X-ray microbeam and analyzing the spot positions in the resulting local Laue pattern. Mapping local hydrostatic strain is usually slower, owing to the need to alternate between white and tunable-energy monochromatic microbeams. A technique has been developed to measure hydrostatic strain while keeping the white beam. The energy of one of the Laue spots of the grain of interest is measured using an energy-dispersive point detector, while simultaneously recording the Laue pattern on the two-dimensional detector. The experimental spot energy, E-exp, is therefore measured at the same time as E-theor, the theoretical spot energy for zero hydrostatic strain, which is derived from the analysis of the Laue pattern. The performance of the technique was compared with that of the monochromatic beam technique in two test cases: a Ge single crystal and a micrometre-sized UO2 grain in a polycrystal. Accuracies on the hydrostatic strain ?a/a of ± 0.4 * 10-4 and ± 1.3 * 10-4 were obtained for Ge and UO2, respectively. Measurement strategies to limit the remaining uncertainties on E-theor are discussed. (authors)

  19. Incidence and common locations of Mongolian spots in newborns: a university hospital

    Directory of Open Access Journals (Sweden)

    Shajari H

    2007-11-01

    Full Text Available Background: Mongolian spots are the most frequently encountered pigmented lesions in newborns. The patches appear at birth or shortly there after, rarely later the MS in term newborns in always present at birth. The shape of MS was commonly either irregular or indefinite, with its borders gradually blending with the surrounding skin. The color most frequently observed in all ethnic groups was blue- green. For the Negro population the color was commonly greenish- blue the next most common color in the total population was blue- gray. Brown coloration in the form of brown specks on a back ground of blue was present in ten percent Negro Newborns. The most common location is the sacra- gluteal region, which frequently is the only part affected. MS occasionally are found in the extremities in those cases with extensive involvement, particularly in the shoulders. The presence of MS in the head or neck has been called aberrant Mongolian spot. The macula has been variously described as irregularly round, oval, roughly triangular, heart shaped, resembling a tennis racket, and angular. The size may vary from a dot of a few millimeters to six or more centimeters in diameter the mark of ten disappears during the first or second year of life. Those marks distant from the sacral region are said to be more apt to persist than the typical sacral one and the buttocks was the site of predilection. Its incidence varies from over 80% in Asians (Mongolian and Chinese to 10% of white infants. Only a limited number of studies were carried out in Iran. Our objective was to study Mongolian spots incidence and common locations in newborns at Shariati hospital."nMethods: During 2004-06, 2305 consecutive newborns were examined at Shariati hospital. Diagnosis of Mongolian spot was based on clinical impression with Pediatricians."nResults: Mongolian spot was observed in 11.4% neonates. The most frequent site of involvement is the sacral, followed by the gluteal area. Mongolian spot did not show a significant relationship to sex, gestational age, mother's age groups and delivery type (p>0.05 but the relationship between Mongolian spot and birth weight groups was significant (p<0.05."nConclusion: Incidence of mongolian spots in our patients was similar to white races. Anthropologic study is necessary for patients admitting to our hospital.

  20. ASTRO-H White Paper - White Dwarf

    OpenAIRE

    Mukai, K.; Yuasa, T.; Harayama, A.; Hayashi, T.; Ishida, M.; Long, K. S.; Terada, Y.; Tsujimoto, M.; Group, On Behalf Of The Astro-h Science Working

    2014-01-01

    Interacting binaries in which a white dwarf accretes material from a companion --- cataclysmic variables (CVs) in which the mass loss is via Roche-lobe overflow, and symbiotic stars in which the white dwarf captures the wind of a late type giant --- are relatively commonplace. They display a wide range of behaviors in the optical, X-rays, and other wavelengths, which still often baffles observers and theorists alike. They are likely to be a significant contributor to the Gal...

  1. Does an Average White Dwarf Have Enough Mass to Prevent an Accretion Disk Tilt?

    OpenAIRE

    Montgomery, M. M.

    2010-01-01

    In a recent publication, we introduce the lift force as a common source to accretion disk tilt that is likely relevant to accretion disk systems. Lift is generated by slightly different supersonic gas stream speeds flowing over and under the disk at the bright spot. In this conference proceeding, we focus on whether the average white dwarf has enough mass to prevent a disk tilt in non-magnetic Cataclysmic Variables (CVs) with accretion disks. Assuming a white dwarf mass of 0...

  2. Connections between Tilted Accretion Disks around White Dwarfs and Substellar Companions

    OpenAIRE

    Montgomery, M. M.

    2011-01-01

    Accretion disks in white dwarf systems are believed to be tilted. In a recent publication, the lift force has been suggested to be a source to disk tilt, a source that is likely relevant to all accretion disk systems. Lift is generated by slightly different supersonic gas stream speeds flowing over and under the disk at the bright spot. In this conference proceeding, we focus on whether a brown dwarf donor star accreting onto a white dwarf primary has enough mass to contribu...

  3. Modelling turbulent spots in swept boundary layers

    International Nuclear Information System (INIS)

    Highlights: • A linear perturbation method can capture the important flow features within a turbulent spot. • The horseshoe vortex in the perturbed velocity field is the dominant flow feature. • Sweep leads to skewing of the turbulent spot and calmed region. • The effects of pressure gradient are generally reduced by sweep. -- Abstract: A computational technique is presented for determining the fully 3-d viscid unsteady perturbation to a non-developing laminar swept boundary layer. For zero pressure gradient, unswept boundary layers, the perturbation method reveals a strongly three dimensional flow within the turbulent spot and its associated calmed region which is very similar to that observed in experiments and full DNS calculations. The perturbation method cannot predict turbulent motion but nevertheless provides a simple yet accurate means of studying and understanding the development of turbulent spot geometry. The most influential flow feature is the horseshoe vortex observed in the fluctuation velocity field, which is responsible for delivering the fluid found in the calmed region between its trailing legs. The upwards flow around the outer periphery of the vortex is also responsible for delivering low momentum fluid to the spot, but additional high momentum fluid also enters the spot from its rear through the downward sweeping motion of fluid between the vortex legs. The effect of an adverse streamwise pressure gradient is to increase the size of the spot and calmed region whereas a favourable pressure gradient has the opposite effect. When sweep is introduced to the boundary layer the spot is skewed for all non-zero pressure gradients, but the changes in size of the spot and calmed region due to pressure gradient are reduced. For favourable pressure gradients the skew increases monotonically with sweep, but this is not the case for adverse pressure gradients where the effect of sweep is more complex

  4. Resistance Spot Welding of dissimilar Steels

    OpenAIRE

    Ladislav Kola?ík; Miroslav Sahul; Marie Kola?íková; Martin Sahul; Milan Tur?a; Michal Felix

    2012-01-01

    This paper presents an analysis of the properties of resistance spot welds between low carbon steel and austenitic CrNi stainless steel. The thickness of the welded dissimilar materials was 2 mm. A DeltaSpot welding gun with a process tape was used for welding the dissimilar steels. Resistance spot welds were produced with various welding parameters (welding currents ranging from 7 to 8 kA). Light microscopy, microhardness measurements across the welded joints, and EDX analysis were used to e...

  5. Clinical anatomy of the G-spot.

    Science.gov (United States)

    Pan, Shu; Leung, Cynthia; Shah, Jaimin; Kilchevsky, Amichai

    2015-04-01

    The existence of the G-Spot has never been unequivocally confirmed. With increased public exposure and a trend towards sexual gratification, however, the impetus to elucidate this structure is greater than ever. This review will focus on research that has been conducted on the clinical anatomy of the G-Spot. Ultimately this review will show that while the distal area of the anterior vaginal wall appears to be the most sensitive region of the vagina, the existence of an anatomical "G-spot" remains to be demonstrated. PMID:25740385

  6. CMB Cold Spot from Inflationary Feature Scattering

    CERN Document Server

    Wang, Yi

    2015-01-01

    We propose a "feature-scattering" mechanism to explain the cosmic microwave background cold spot seen from {\\it WMAP} and {\\it Planck} maps. If there are hidden features in the potential of multi-field inflation, the inflationary trajectory can be scattered by such features. The scattering is controlled by the amount of isocurvature fluctuations, and thus can be considered as a mechanism to convert isocurvature fluctuations into curvature fluctuations. This mechanism predicts localized cold spots (instead of hot ones) on the CMB. In addition, it may also bridge a connection between the cold spot and a dip on the CMB power spectrum at $\\ell \\sim 20$.

  7. Digital Sun Sensor Multi-Spot Operation

    OpenAIRE

    Michele Grassi; Giancarlo Rufino

    2012-01-01

    The operation and test of a multi-spot digital sun sensor for precise sun-line determination is described. The image forming system consists of an opaque mask with multiple pinhole apertures producing multiple, simultaneous, spot-like images of the sun on the focal plane. The sun-line precision can be improved by averaging multiple simultaneous measures. Nevertheless, the sensor operation on a wide field of view requires acquiring and processing images in which the number of sun spots and the...

  8. Virus Diseases of Cucurbits in Gaziantep-Turkey

    Directory of Open Access Journals (Sweden)

    Mehmet Ozaslan

    2006-01-01

    Full Text Available Cucurbit growing is affected negatively due to diseases caused by cucurbit viruses. In order to prevent this damage cucurbit viruses were identified by serologically. Due to this study, it is usually difficult to give definitive diagnosis based on symptoms but occasionally symptoms; are curling, wrinkling, spot mosaics, yellowing, shape deformation on leaves, smaller leaves than normal, buff-colored mosaics, observed on younger leaves of cucurbits and stunting , distortion and fruit deformation on the plants. After this, samples collected and taken to laboratory to determine the virus which caused this symptoms and DAS -ELISA tests were performed to determine CMV (Cucumber mosaic virus, CABYV (Cucurbit aphid borne yellow mosaic virus, ZYMV (Zucchini yellow mosaic virus, ToMV (Tomato mosaic virus, PMMV (Pepper mild mottle virus, PXV (Potato X virus, PYV (Potato Y virus on the samples collected during July-August in 2004. At the end of this test out of 56 samples, 10 were found to be infected with one or more virus. As a result of this study 20 samples were infected by CMV and 22 samples were infected by ZYMV and 3 samples were infected by PVY.

  9. Kalanchoë blossfeldiana, a new host for Sonchus yellow net virus

    OpenAIRE

    Bouwen, I.; Schoen, C. D.; Balen, E.; Vlugt, R. A. A.

    2002-01-01

    The agent causing chlorotic spots in Kalanchoë blossfeldiana `Isabella¿ was investigated. A virus isolated from this naturally infected kalanchoë was mechanically transmissible to several indicator plants. Observation of suspension preparations in the electron microscope revealed rhabdovirus-like particles. On the basis of symptoms on indicator plants, serology, electron microscopy, molecular characterisation and back inoculation to K. blossfeldiana 'Isabella', the causal agent was identif...

  10. Accreting White Dwarfs

    OpenAIRE

    Hernanz, Margarita; Jose?, Jordi

    2008-01-01

    Thermonuclear (type Ia) supernovae are explosions in accreting white dwarfs, but the exact scenario leading to these explosions is still unclear. An important step to clarify this point is to understand the behaviour of accreting white dwarfs in close binary systems. The characteristics of the white dwarf (mass, chemical composition, luminosity), the accreted material (chemical composition) and those related with the properties of the binary system (mass accretion rate), are...

  11. White matter dementia

    OpenAIRE

    Filley, Christopher M.

    2012-01-01

    White matter dementia (WMD) is a syndrome introduced in 1988 to highlight the potential of cerebral white matter disorders to produce cognitive loss of sufficient severity to qualify as dementia. Neurologists have long understood that such a syndrome can occur, but the dominance of gray matter as the locus of higher function has strongly directed neurobehavioral inquiry to the cerebral cortex while white matter has received less attention. Contemporary neuroimaging has been crucial in enablin...

  12. Physical white chaos generation

    OpenAIRE

    Wang, Anbang; Wang, Yuncai; Wang, Bingjie; Li, Lei; Zhang, Mingjiang; Zhang, Wendong

    2014-01-01

    Physical chaos is a fascinating prospect for high-speed data security by serving as a masking carrier or a key source, but suffers from a colored spectrum that divulges system's intrinsic oscillations and degrades randomness. Here, we demonstrate that physical chaos with a white spectrum can be achieved by the optical heterodyning of two delayed-feedback lasers. A white chaotic spectrum with 1-dB fluctuation in a band of 11 GHz is experimentally obtained. The white chaos als...

  13. White Men Write Now

    OpenAIRE

    Manson, Richard

    2005-01-01

    Contemporary literature by white men in the United States about the identity "white man" largely focuses on two themes: destabilized structures of identity and their reinforcement. This concern with the threat to identity structures of race and gender is an old one in American literature. However, it takes on a new meaning in the present postmodern climate that challenges master narratives and their reinscription by forces which sustain them. White Men Write Now repeatedly returns to the thes...

  14. Price dynamics in electricity spot markets

    OpenAIRE

    Florentina Paraschiv; Michael Schürle

    2013-01-01

    We propose a novel regime-switching approach for the simulation of electricity spot prices that is inspired by the class of fundamental models and takes into account the relation between spot and forward prices. Additionally the model is able to reproduce spikes and negative prices. Market prices are derived given an observed forward curve. We distinguish between a base regime and an upper as well as a lower spike regime. The model parameters are calibrated using historical hourly price forwa...

  15. X-ray spot film device

    International Nuclear Information System (INIS)

    Improvements are described in an X-ray spot film device which is used in conjunction with an X-ray table to make a selected number of radiographic exposures on a single film and to perform fluoroscopic examinations. To date, the spot film devices consist of two X-ray field defining masks, one of which is moved manually. The present device is more convenient to use and speeds up the procedure. (U.K.)

  16. Actor based framework for SunSPOTs

    OpenAIRE

    Askirk, David Christian

    2009-01-01

    This project covers the development of an actor-based framework for the SunSPOT platform, a wireless sensor from Sun Microsystems. It covers the possibility for implementing a framework, given a hardware unit with built in communication stack. Different languages will be compared to the actor definition and whether they can be used on the J2ME platform. Implementation of load balancing will be attempted. Communication on the SunSPOT is tested and the results show that once a...

  17. Concentrator hot-spot testing, phase 1

    Science.gov (United States)

    Gonzalez, C. C.

    1987-01-01

    Results of a study to determine the hot-spot susceptibility of concentrator cells, to provide a hot-spot qualification test for concentrator modules, and to provide guidelines for reducing hot-spot susceptibility are presented. Hot-spot heating occurs in a photovoltaic module when the short-circuit current of a cell is lower than the string operating current forcing the cell into reverse bias with a concurrent power dissipation. Although the basis for the concentrator module hot-spot qualification test is the test developed for flat-plate modules, issues, such as providing cell illumination, introduce additional complexities into the testing procedure. The same general guidelines apply for protecting concentrator modules from hot-spot stressing as apply to flat-plate modules. Therefore, recommendations are made on the number of bypass diodes required per given number of series cells per module or source circuit. In addition, a new method for determining the cell temperature in the laboratory or in the field is discussed.

  18. MR imaging of white matter lesions in AIDS

    International Nuclear Information System (INIS)

    Autopsy reports have shown white-matter abnormalities from infection of the brain by the human immunodeficiency virus (HIV), the agent that causes acquired immunodeficiency syndrome (AIDS). The authors observed abnormal signal on T2-weighted images in the white matter of approximately one third of all AIDS patients. Of 50 patients with white-matter lesions, approximately two thirds had no clinical or biopsy evidence of cytomegalovirus, toxoplasmosis, PML, or lymphoma. Several patients were shown at autopsy to have isolated evidence of HIV encephalitis. The authors conclude that white-matter lesions are common in AIDS and are frequently caused by infection with HIV. Some MR findings may be helpful in characterizing these lesions, but the various etiologies are often indistinguishable

  19. HIV virus

    Science.gov (United States)

    Carl Henderson (National Institutes of Health; )

    2005-12-09

    HIV is a virus that can be transmitted through fluids exchanged in sexual activity. HIV eventually causes AIDS. AIDS patients have compromised immune systems and they eventually die from diseases that healthy humans would normally fight off very easily.

  20. TAT improves in vitro transportation of fortilin through midgut and into hemocytes of white shrimp Litopenaeus vannamei

    Science.gov (United States)

    Zhou, Yi; Zhang, Wenbing; Mai, Kangsen; Xu, Wei; Zhang, Yanjiao; Ai, Qinghui; Wang, Xiaojie

    2012-06-01

    Fortilin is a multifunctional protein implicated in many important cellular processes. Since injection of Pm-fortilin reduces shrimp mortality caused by white spot syndrome virus (WSSV), there is potential application of fortilin in shrimp culture. In the present study, in order to improve trans-membrane transportation efficiency, the protein transduction domain of the transactivator of transcription (TAT) peptide was fused to fortilin. The Pichia pastoris yeast expression system, which is widely accepted in animal feeds, was used for production of recombinant fusion protein. Green fluorescence protein (GFP) was selected as a reporter because of its intrinsic visible fluorescence. The fortilin, TAT and GFP fusion protein were constructed. Their trans-membrane transportation efficiency and effects on immune response of shrimp were analyzed in vitro. Results showed that TAT peptide improved in vitro uptake of fortilin into the hemocytes and midgut of Litopenaeus vannamei. The phenoloxidase (PO) activity of hemocytes incubated with GFP-Fortilin or GFP-Fortilin-TAT was significantly increased compared with that in the control without expressed fortilin. The PO activity of hemocytes incubated with 200 ?g mL-1 GFP-Fortilin-TAT was significantly higher than that in the group with the same concentration of GFP-Fortilin. Hemocytes incubated with GFP-Fortilin-TAT at all concentrations showed significantly higher nitric oxide synthase (NOS) activity than those in the control or in the GFP-Fortilin treatment. The present in vitro study indicated that TAT fusion protein improved the immune effect of fortilin.

  1. Carbon in white dwarfs

    International Nuclear Information System (INIS)

    The problem of obtaining a considerable carbon enhancement in nova envelopes to account for the occurrence of fast novae is examined in detail. No mechanism seems completely satisfactory from the point of view of stellar evolution and white dwarf structure, even considering evidence from single white dwarf observations

  2. Ryan White Program

    Science.gov (United States)

    ... legislation_hist.htm. ? Return to text Ryan White HIV/AIDS Treatment Modernization Act of 2006 (P.L. 109-415). ? Return to text Ryan White HIV/AIDS Treatment Extension Act of 2009 (P.L. 111-87). ? ...

  3. White Teachers Talking Race

    Science.gov (United States)

    Segall, Avner; Garrett, James

    2013-01-01

    In light of the increasing racial diversity in American schools and the consistently homogenous teacher workforce in the United States, understanding the ways white teachers consider and attend to racial issues is of crucial importance to the educational landscape. This paper, based on a qualitative study, explores five white American…

  4. SPOTTED STAR LIGHT CURVES WITH ENHANCED PRECISION

    International Nuclear Information System (INIS)

    The nearly continuous timewise coverage of recent photometric surveys is free of the large gaps that compromise attempts to follow starspot growth and decay as well as motions, thereby giving incentive to improve computational precision for modeled spots. Due to the wide variety of star systems in the surveys, such improvement should apply to light/velocity curve models that accurately include all the main phenomena of close binaries and rotating single stars. The vector fractional area (VFA) algorithm that is introduced here represents surface elements by small sets of position vectors so as to allow accurate computation of circle-triangle overlap by spherical geometry. When computed by VFA, spots introduce essentially no noticeable scatter in light curves at the level of one part in 10,000. VFA has been put into the Wilson-Devinney light/velocity curve program and all logic and mathematics are given so as to facilitate entry into other such programs. Advantages of precise spot computation include improved statistics of spot motions and aging, reduced computation time (intrinsic precision relaxes needs for grid fineness), noise-free illustration of spot effects in figures, and help in guarding against false positives in exoplanet searches, where spots could approximately mimic transiting planets in unusual circumstances. A simple spot growth and decay template quantifies time profiles, and specifics of its utilization in differential corrections solutions are given. erential corrections solutions are given. Computational strategies are discussed, the overall process is tested in simulations via solutions of synthetic light curve data, and essential simulation results are described. An efficient time smearing facility by Gaussian quadrature can deal with Kepler mission data that are in 30 minute time bins.

  5. Spotted fever group rickettsial infection in dogs from eastern Arizona: how long has it been there?

    Science.gov (United States)

    Nicholson, William L; Gordon, Rondeen; Demma, Linda J

    2006-10-01

    A serosurvey of free-roaming dogs for antibodies to spotted fever group rickettsiae was conducted using archival samples that had been collected in the White Mountain region of eastern Arizona during a plague study in 1996. Immunoglobulin G antibodies to Rickettsia rickettsii (5.1%) and to R. rhipicephali (3.6%) were demonstrated, and no cross-reactive samples were identified. This study indicates that R. rickettsii was likely present in the dog populations in this area prior to the recognition of human cases of Rocky Mountain spotted fever (RMSF). The role of dogs as short-term reservoirs and primary hosts for the vector tick, Rhipicephalus sanguineus, should receive closer attention. PMID:17114769

  6. Computer Viruses. Technology Update.

    Science.gov (United States)

    Ponder, Tim, Comp.; Ropog, Marty, Comp.; Keating, Joseph, Comp.

    This document provides general information on computer viruses, how to help protect a computer network from them, measures to take if a computer becomes infected. Highlights include the origins of computer viruses; virus contraction; a description of some common virus types (File Virus, Boot Sector/Partition Table Viruses, Trojan Horses, and…

  7. Dried blood spot UHPLC-MS/MS analysis of oseltamivir and oseltamivircarboxylate-a validated assay for the clinic

    OpenAIRE

    Hooff, G.P.; Meesters, R. J. W.; Kampen, J.J.A. van; Huizen, N.A. van; Koch, B.; Al Hadithy, A F Y; Gelder, T., van; Osterhaus, A. D. M. E.; Gruters, R. A.; Luider, T. M.

    2011-01-01

    The neuraminidase inhibitor oseltamivir (Tamiflu®) is currently the first-line therapy for patients with influenza virus infection. Common analysis of the prodrug and its active metabolite oseltamivircarboxylate is determined via extraction from plasma. Compared with these assays, dried blood spot (DBS) analysis provides several advantages, including a minimum sample volume required for the measurement of drugs in whole blood. Samples can easily be obtained via a simple, non-invasive finger ...

  8. LEOPARD syndrome: what are café noir spots?

    Science.gov (United States)

    Rodríguez-Bujaldón, Alfonso; Vazquez-Bayo, Carmen; Jimenez-Puya, Rafael; Galan-Gutierrez, Manuel; Moreno-Gimenez, José; Rodriguez-Garcia, Alfonso; Tercedor, Jesus; Velez-Garcia, Antonio

    2008-01-01

    Lentigines, electrocardiographic abnormalities, ocular hypertelorism, pulmonary stenosis, abnormalities of genitalia, retarded growth, and deafness syndrome (multiple lentigines syndrome) is most often characterized by multiple lentigines and cardiac conduction defects. Café noir spot is a term proposed, by analogy to café au lait spots, for the larger and darkly pigmented patches that are frequently observed in patients with this syndrome. Although presumed by some authors to represent lentigines, the histologic features of café noir spots have not been well documented in the literature. Only two previous cases have been reported in which a biopsy of the café noir spots than melanocytic nevi. We describe the histologic characteristics of seven café noir spots in six patients with lentigines, electrocardiographic abnormalities, ocular hypertelorism, pulmonary stenosis, abnormalities of genitalia, retarded growth, and deafness syndrome. Three lesions represented melanocytic nevi (one with dysplastic features), and four were compatible with lentigo simplex. These findings help our understanding of the histologic spectrum of pigmented lesions in lentigines, electrocardiographic abnormalities, ocular hypertelorism, pulmonary stenosis, abnormalities of genitalia, retarded growth, and deafness syndrome. PMID:18789084

  9. 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 (<~0.03 Hz), may enhance the 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.

  10. Autoradiographic localization of the synthetic sites of tomato spoted wilt virus and potato virus Y

    International Nuclear Information System (INIS)

    The biosynthesis sites were investigated of two morfologically different viruses - the Tomato Spotted Wilt Virus (TSWV-spherical particle) and the Potato Virus Y (PVY - long and flexuous particle) in order to discuss the hypothesis of De Zoeten and Schlegel about the relationship between virus morphology and the location of the viral biosynthesis. Samples from uninfected or infected leaves were immersed in distilled water or an aqueous solution and transfered to uridine tritiated solution. After washing in distilled water the samples were fixed, dehydrated and embedded in Epon 812 for electron microscopy conventional techniques. Ultrathin sections were covered with Ilford L-4 photographic emulsion and exposed for two months before photographic development, staining and examinated in the electron microscope. The number of silver grains per unit areas (grain density) in the electronphotomicrographs was used to compare the grains densities of some cells regions of tissues treated or not with AMD. The result indicated the endoplasmic reticulum as the most likely location of the TSWV-RNA replication. The same comparison made with tobacco cells infected with PVY showed that the cytoplasmic area is the most probable site of the PVY-RNA replication. The results obtained seem to show that the rule proposed by De Zoeten and Schlegel cannot be used for all plant viruses because the TSWV replicates in the cytoplasm of infected cell. These viruses seem to be exceptions to that rul viruses seem to be exceptions to that rule. (Author)

  11. A combined oligonucleotide and protein microarray for the codetection of nucleic acids and antibodies associated with human immunodeficiency virus, hepatitis B virus, and hepatitis C virus infections.

    Science.gov (United States)

    Perrin, Agnès; Duracher, David; Perret, Magali; Cleuziat, Philippe; Mandrand, Bernard

    2003-11-15

    A multiplexed assay based on the codetection of nucleic acids and antibodies in human serum infected by human immunodeficiency virus (HIV), hepatitis B virus (HBV) or hepatitis C virus was proposed. The combined immuno- and oligosorbent array (CombOLISA) microarray is prepared in 96-well standard microplates by spotting (1). nucleic probes specific for a virus genome, (2). viral proteins for the capture of serum antibodies, and (3). nonspecific proteins for verifying specificity. Experimental assay conditions were optimized so that both DNA hybridization and immunological reactions can be achieved simultaneously in the same well and buffer and all at the same temperature. A generic detection system based on the precipitation of an insoluble colorimetric substrate in the presence of enzyme-labeled antibodies or streptavidin was proposed. The optical density of each spot was correlated to the corresponding analyte concentration. The influence of critical parameters on CombOLISA performance such as serum concentration was studied. Calibration curves and sensitivity thresholds were established for each parameter. Serial dilutions of serum were correlated to results obtained with validated immunoassay platforms such as a microplate enzyme-linked immunosorbent assay or the VIDAS automat. Also, several HIV- and HBV-infected serum samples were tested independently by CombOLISA and VIDAS. Coefficients of variation for genomic and proteomic parameters vs spot density were below 15%. PMID:14596821

  12. Zeeman tomography of magnetic white dwarfs, I. Reconstruction of the field geometry from synthetic spectra

    CERN Document Server

    Euchner, F; Beuermann, K; Gänsicke, B T; Hessman, F V

    2002-01-01

    We have computed optical Zeeman spectra of magnetic white dwarfs for field strengths between 10 and 200MG and effective temperatures between 8000 and 40000K. They form a database containing 20628 sets of flux and circular polarization spectra. A least-squares optimization code based on an evolutionary strategy can recover relatively complex magnetic field topologies from phase-resolved synthetic Zeeman spectra of rotating magnetic white dwarfs. We consider dipole and quadrupole components which are non-aligned and shifted off-centre. The model geometries include stars with a single high-field spot and with two spots separated by approx. 90 degrees. The accuracy of the recovered field structure increases with the signal-to-noise ratio of the input spectra and is significantly improved if circular polarization spectra are included in addition to flux spectra. We discuss the strategies proposed so far to unravel the field geometries of magnetic white dwarfs.

  13. Molecular interactions between hepatitis B virus and delta virus

    Science.gov (United States)

    Shirvani-Dastgerdi, Elham; Tacke, Frank

    2015-01-01

    As a deficient virus due to the lack of envelope proteins, hepatitis D virus (HDV) causes chronic or fulminant “delta hepatitis” only in people with simultaneous hepatitis B virus (HBV) infection. HBV encodes three types of surface proteins known as small (S), medium (M) and large (L) envelope proteins. All three types of HBV surface antigens (HBsAgs) are present on HDV virions. The envelopment process of HDV occurs through interactions between the HDV ribonucleoprotein (RNP) complex and HBV HBsAgs. While HBsAg is the only protein required by HDV, the exact interaction sites between the S protein and pre-mature HDV are not well defined yet. In fact, these sites are distributed along the S protein with some hot spots for the envelopment process. Moreover, in most clinically studied samples, HDV infection is associated with a dramatically reduced HBV viral load, temporarily or permanently, while HBsAg resources are available for HDV packaging. Thus, beyond interacting with HBV envelope proteins, controlling mechanisms exist by which HDV inhibits HBV-DNA replication while allowing a selective transcription of HBV proteins. Here we discuss the molecular interaction sites between HBsAg and the HDV-RNP complex and address the proposed indirect mechanisms, which are employed by HBV and HDV to facilitate or inhibit each other’s viral replication. Understanding molecular interactions between HBV and HDV may help to design novel therapeutic strategies for delta hepatitis. PMID:25964870

  14. Simulating acoustic waves in spotted stars

    CERN Document Server

    Papini, Emanuele; Gizon, Laurent; Hanasoge, Shravan M

    2015-01-01

    Acoustic modes of oscillation are affected by stellar activity, however it is unclear how starspots contribute to these changes. Here we investigate the non-magnetic effects of starspots on global modes with angular degree $\\ell \\leq 2$ in highly active stars, and characterize the spot seismic signature on synthetic light curves. We perform 3D time-domain simulations of linear acoustic waves to study their interaction with a model starspot. We model the spot as a 3D change in the sound speed stratification with respect to a convectively stable stellar background, built from solar Model S. We perform a parametric study by considering different depths and perturbation amplitudes. Exact numerical simulations allow investigation of the wavefield-spot interaction beyond first order perturbation theory. The interaction of the axisymmetric modes with the starspot is strongly nonlinear. As mode frequency increases, the frequency shifts for radial modes exceed the value predicted by linear theory, while the shifts for...

  15. White Pratt from Luton

    OpenAIRE

    White, Laura; Allan, Dominic; Pratt, Robert

    2011-01-01

    Three person exhibition. 34 Toynebee Street, London. 'White Pratt From Luton' landed on No.34 Toynebee Street on Friday 24th June, the moment Dominic Allan took receipt of keys from East End Homes. A four-week contract, four walls and a sink. Half decent lighting, peeling paint and raw plugs showing their arses. A space that’s given everything of itself to artists Laura White, Robert Pratt and Dominic Allan. A bespoke A3 lightbox sits in the window. ‘White Pratt From Luton’ is c...

  16. Analysis of reproduction and litter performance of the Zlotnicka Spotted breed and its different crossbreeds

    OpenAIRE

    D Stanis?awski; Zbigniew Sobek; Anna Jankowska; Damian Knecht; Anna Panek; Ewa Skrzypczak; Karolina Szulc

    2011-01-01

    Zlotnicka Spotted (ZS) is a native Polish pig breed. It is characterized by low reproduction and litter performance. This study aimed to determine the suitability of ZS pigs when crossed with Duroc and Polish Large White (PLW) according to reproduction and litter performance. Data concerning the number of piglets in the litter, litter weight, litter balance and piglet wastage were evaluated at birth, on the 7th and 21st days, and during weaning. ZS sows in purebred litters had 8.82 piglets at...

  17. Volatility modelling of CO2 spot prices

    OpenAIRE

    Benschop, Thijs

    2013-01-01

    In this paper we analyse the short-term spot price of European Union Allowances (EUAs), which is of particular importance in the transition of energy markets and for the development of new risk management strategies. We use daily spot market data from the second trading period of the EU ETS. Emphasis is given to short-term forecasting of prices and volatility. Due to the characteristics of the price process, such as volatility modelling, breaks in the volatility process and heavy-tailed ...

  18. Statistical Hot Spot Model for Explosive Detonation

    Energy Technology Data Exchange (ETDEWEB)

    Nichols III, A L

    2004-05-10

    The Non-local Thermodynamic Equilibrium Statistical Hot Spot Model (NLTE SHS), a new model for explosive detonation, is described. In this model, the formation, ignition, propagation, and extinction of hot spots is explicitly modeled. The equation of state of the explosive mixture is treated with a nonlocal equilibrium thermodynamic assumption. A methodology for developing the parameters for the model is discussed, and applied to the detonation velocity diameter effect. Examination of these results indicates where future improvements to the model can be made.

  19. Photodynamic treatment of Herpes simplex virus infection in vitro

    International Nuclear Information System (INIS)

    The effects of photodynamic action on in vitro herpes simplex virus infections of CV-1 monkey kidney fibroblasts or human skin fibroblasts were determined using proflavine sulfate and white fluorescent lamps. Photodynamic treatment of confluent cell monolayers prior to virus infection inactivated cell capacity, i.e. the capacity of the treated cells to support subsequent virus growth as measured by plaque formation. The capacity of human cells was more sensitive to inactivation than the capacity of monkey cells when 6 ?M proflavine was used. Treated cell monolayers recovered the capacity to support virus plaque formation when virus infection was delayed four days after the treatment. Experiments in which the photodynamically treated monolayers were infected with UV-irradiated virus demonstrated that this treatment induced Weigle reactivation in both types of cells. This reactivation occurred for virus infection just after treatment or 4 days later. A Luria-Latarjet-type experiment was also performed in which cultures infected with unirradiated virus were photodynamically treated at different times after the start of infection. The results showed that for the first several hours of the virus infection the infected cultures were more sensitive to inactivation by photodynamic treatment than cell capacity. By the end of the eclipse period the infected cultures were less sensitive to inactivation than cell capacity. Results from extracellular inactivation of virus growth tracellular inactivation of virus growth in monkey cells at 6 ?M proflavine indicated that at physiological pH the virus has a sensitivity to photodynamic inactivation similar to that for inactivation of cell capacity. The combined data indicated that photodynamic treatment of the cell before or after virus infection could prevent virus growth. Thus, photodynamic inactivation of infected and uninfected cells may be as important as inactivation of virus particles when considering possible mechanisms in clinical photodynamic therapy for herpes simplex. (author)

  20. New Experimental Hosts of Tobacco streak virus and Absence of True Seed Transmission in Leguminous Hosts

    OpenAIRE

    Vemana, K.; Jain, R. K.

    2010-01-01

    Of 70 plant species tested, 50 species were susceptible to Tobacco streak virus (TSV) on sap inoculation. Both localized (necrotic and chlorotic spots) and systemic (necrotic spots, axillary shoot proliferation, stunting, total necrosis and wilt) symptoms are observed by majority of plant species. Eleven new experimental hosts were identified viz., Amaranthus blitum var. oleracea (Chaulai sag), Celosia cristata (Cocks comb), Beta vulgaris var. bengalensis (Palak/Indian spinach), Calendula off...