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Sample records for white spot virus

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

    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.

  2. Draft Genome Sequence of White Spot Syndrome Virus Isolated from Cultured Litopenaeus vannamei in Mexico

    Rodriguez-Anaya, Libia Zulema; Gonzalez-Galaviz, Jose Reyes; Casillas-Hernandez, Ramón; Lares-Villa, Fernando; Estrada, Karel

    2016-01-01

    The first genome sequence of a Mexican white spot syndrome virus is presented here. White spot syndrome is a shrimp pandemic virus that has devastated production in Mexico for more than 10 years. The availability of this genome will greatly aid epidemiological studies worldwide, contributing to the molecular diagnostic and disease prevention in shrimp farming. PMID:26966222

  3. Proteomic Analyses of the Shrimp White Spot Syndrome Virus

    Yan-wei TAN; Zheng-li SHI

    2008-01-01

    White spot syndrome virus (WSSV), a unique member within the virus family Nimaviridae, is the most notorious aquatic virus infecting shrimp and other crustaceans and has caused enormous economic losses in the shrimp farming industry worldwide. Therefore, a comprehensive understanding of WSSV morphogenesis, structural proteins, and replication is essential for developing prevention measures of this serious parasite. The viral genome is approximately 300kb and contains more than 180 open reading frames (ORF). However, most of proteins encoded by these ORF have not been characterized. Due to the importance of WSSV structural proteins in the composition of the virion structure, infection process and interaction with host cells, knowledge of structural proteins is essential to understanding WSSV entry and infection as well as for exploring effective prevention measures. This review article summarizes mainly current investigations on WSSV structural proteins including the relative quantities, localization, function and protein-protein interactions. Traditional proteomic studies of 1D or 2D gel electrophoresis separations and mass spectrometry (MS) followed by database searches have identified a total of 39 structural proteins. Shotgun proteomics and iTRAQ were initiated to identify more structural proteins. To date, it is estimated that WSSV is assembled by at least 59 structural proteins, among them 35 are defined as the envelope fraction (including tegument proteins) and 9 as nucleocapsid proteins. Furthermore, the interaction within several major structural proteins has also been investigated. This identitification and characterization of WSSV protein components should help in the understanding of the viral assembly process and elucidate the roles of several major structural proteins.

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

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

    2015-03-06

    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.

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

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

    2000-01-01

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

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

    Hulten, van M.C.W.

    2001-01-01

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

  7. White spot syndrome virus epizootic in cultured Pacific white shrimp Litopenaeus vannamei (Boone) in Taiwan.

    Cheng, L; Lin, W-H; Wang, P-C; Tsai, M-A; Hsu, J-P; Chen, S-C

    2013-12-01

    White spot syndrome virus (WSSV) has caused significant losses in shrimp farms worldwide. Between 2004 and 2006, Pacific white shrimp Litopenaeus vannamei (Boone) were collected from 220 farms in Taiwan to determine the prevalence and impact of WSSV infection on the shrimp farm industry. Polymerase chain reaction (PCR) analysis detected WSSV in shrimp from 26% of farms. Juvenile shrimp farms had the highest infection levels (38%; 19/50 farms) and brooder shrimp farms had the lowest (5%; one of 20 farms). The average extent of infection at each farm was as follows for WSSV-positive farms: post-larvae farms, 71%; juvenile farms, 61%; subadult farms, 62%; adult farms, 49%; and brooder farms, 40%. Characteristic white spots, hypertrophied nuclei and basophilic viral inclusion bodies were found in the epithelia of gills and tail fans, appendages, cephalothorax and hepatopancreas, and virions of WSSV were observed. Of shrimp that had WSSV lesions, 100% had lesions on the cephalothorax, 96% in gills and tail fans, 91% on appendages and 17% in the hepatopancreas. WSSV was also detected in copepoda and crustaceans from the shrimp farms. Sequence comparison using the pms146 gene fragment of WSSV showed that isolates from the farms had 99.7-100% nucleotide sequence identity with four strains in the GenBank database--China (AF332093), Taiwan (AF440570 and U50923) and Thailand (AF369029). This is the first broad study of WSSV infection in L. vannamei in Taiwan.

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

    Tsai, M.F.; Lo, C.F.; Hulten, van M.C.W.; 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 e

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

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

    2015-01-01

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

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

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

    2010-01-01

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

  11. Parámetros inmunitaríos en camarones juveniles lv en piscinas camaroneras infectadas con white spot syndrom virus

    Montesdeoca, Bélgica Mercedes; Ibarra, Eulalia; Rodríguez, Jenny

    2002-01-01

    Parámetros inmunitarios en camarones juveniles LV en piscinas camaroneras infectadas con White Spot Syndrom Virus Parámetros inmunitarios en camarones juveniles LV en piscinas camaroneras infectadas con White Spot Syndrom Virus

  12. White spot syndrome virus envelope protein VP124 involved in the virus infection

    ZHU Yanbing; WU Chenglin; YANG Feng

    2008-01-01

    White spot syndrome virus(WSSV)is one of the major shrimp pathogens causing large economic losses to shrimp farming.In an attempt to identify the envelope proteins involved in the virus infection,purified WSSV virions were mixed with three antisera against WSSV envelope proteins(VP39,VP124 and VP187),individually.And then they were injected intramuscularly into crayfish (Procambarus clarkii) to conduct in vivo neutralization assays.The results showed that for groups injected with virions only and groups injected with the mixture of virions and antiserum against VP124,the crayfish mortalities were 100% and 60% on the 8th day postinfection,individually.The virus infection could be delayed or neutralized by antibody against the envelope pro-tein VP124.Quantitative PCR was used to further investigate the influence of three antisera described above on the virus infec-tion.The results showed that the antiserum against VP124 could restrain the propagation of WSSV in crayfish.All of the results suggested that the viral envelope protein VP124 played a role in WSSV infection.

  13. DETECTION OF WHITE SPOT SYNDROME VIRUS(WSSV) OF PENAEUS CHINENSIS BY IN SITU HYBRIDIZATION

    2000-01-01

    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.

  14. Production and Characterization of Monoclonal Antibodies of Shrimp White Spot Syndrome Virus Envelope Protein VP28

    Wan-gang GU; Jun-fa YUAN; Ge-lin XU; Li-juan LI; Ni LIU; Cong ZHANG; Jian-hong ZHANG; Zheng-li SHI

    2007-01-01

    BALB/c mice were immunized with purified White spot syndrome virus (WSSV).Six monoclonal antibody cell lines were selected by ELISA with VP28 protein expressed in E.coll in vitro neutralization experiments showed that 4 of them could inhibit the virus infection in crayfish.Westernblot suggested that all these monoclonal antibodies were against the conformational structure of VP28.The monoclonal antibody 7B4 was labeled with colloidal gold particles and used to locate the VP28 on virus envelope by immunogold labeling.These monoclonal antibodies could be used to develop immunological diagnosis methods for WSSV infection.

  15. Arginine kinase of Litopenaeus vannamei involved in white spot syndrome virus infection.

    Ma, Fang-fang; Liu, Qing-hui; Guan, Guang-kuo; Li, Chen; Huang, Jie

    2014-04-10

    Virus-host interaction is important for virus infection. White spot syndrome virus VP14 contains transmembrane and signal peptides domain, which is considered to be important for virus infection. Until now, the function of this protein remains undefined. In this study, we explored the interaction of VP14 with host cell. A new shrimp protein (arginine kinase of Litopenaeus vannamei, LvAK) is selected and its localization in shrimp cells is also confirmed. Cellular localization of LvAK protein in shrimp hemocytes showed that LvAK was primarily located at the periphery of hemocytes and was scarcely detectable in the nucleus. Tissue distribution indicated that arginine kinase gene was spread commonly in the tissues and was highly present in shrimp muscle tissue. The expression of LvAK mRNA in muscle was significantly up-regulated after WSSV stimulation. Indirect immunofluorescence assay showed that LvAK interacted with VP14 in WSSV-infected shrimp. Injection of LvAK protein enhanced the mortality of shrimp infected with white spot syndrome virus (WSSV). These results showed that LvAK is involved in WSSV infection. Future research on this topic will help to reveal the molecular mechanism of WSSV infection.

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

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

    2014-01-01

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

  17. Assessment of the role of brine shrimp Artemia in white spot syndrome virus (WSSV) transmission.

    Zhang, Jia-Song; Dong, Shuang-Lin; Dong, Yun-Wei; Tian, Xiang-Li; Cao, Yi-Cheng; Li, Zuo-Jia; Yan, Dong-Chun

    2010-01-01

    Challenge tests with Artemia four different development stages (nauplii, metanauplii, pseudoadults and adults) to white spot syndrome virus was carried out by immersion challenge and virus-phytoplankton adhesion route in order to asses the possibility of Artemia acting as a vector of WSSV to penaeid shrimp Litopenaeus vannamei postlarvae. The WSSV succeeded in infecting four stages Artemia, and nested-PCR detection for WSSV revealed positive results to virus-phytoplankton adhesion route. No mass mortalities were observed in penaeid shrimp postlarvae fed with WSSV-positive Artemia which exposed to WSSV by virus-phytoplankton adhesion route, whereas WSSV DNA detected in penaeid shrimp postlarvae by nested-PCR. By contrary, no WSSV-positive was detected in any animal fed with WSSV-negative Artemia. These results indicated that Artemia could serve as a vector in WSSV transmission.

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

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

    2016-01-18

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

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

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

    2017-01-01

    Background White spot disease (WSD), a major threat to sustainable aquaculture worldwide, is caused by White spot syndrome virus (WSSV). The diagnosis of WSD relies heavily on molecular detection of the virus by one-step PCR. These procedures are neither field-usable nor rapid enough considering the speed at which the virus spreads. Thus, development of a rapid, reliable and field-usable diagnostic method for the detection of WSSV infection is imperative to prevent huge economic losses. Methods/Principal Findings Here, we report on the development of a lateral flow immunoassay (LFIA) employing gold nanoparticles conjugated to a polyclonal antibody against VP28 (envelope protein of WSSV). The LFIA detected WSSV in ~20 min and showed no cross-reactivity with other shrimp viruses, viz. Monodon Baculovirus (MBV), Hepatopancreatic parvovirus (HPV) and Infectious Hypodermal and Hematopoietic Necrosis virus (IHHNV). The limit of detection (LOD) of the assay, as determined by real-time PCR, was 103 copies of WSSV. In a time course infectivity experiment, ~104 WSSV particles were injected in Litopenaeus vannamei. The LFIA could rapidly (~ 20 min) detect the virus in different tissues after 3 h (hemolymph), 6 h (gill tissue) and 12 h (head soft tissue, eye stalk, and pleopod) of infection. Based on these findings, a validation study was performed using 75 field samples collected from different geographical locations in India. The LFIA results obtained were compared with the conventional “gold standard test”, viz. one-step PCR. The analysis of results in 2x2 matrix indicated very high sensitivity (100%) and specificity (96.77%) of LFIA. Similarly, Cohen’s kappa coefficient of 0.983 suggested "very good agreement” between the developed LFIA and the conventional one-step PCR. Conclusion The LFIA developed for the rapid detection of WSSV has an excellent potential for use in the field and could prove to be a boon to the aquaculture industry. PMID:28046005

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

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

    2014-01-01

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

  1. On the role of the polychaete Dendronereis spp. i the transmission of white spot syndrome virus in shrimp ponds

    Desrina, Haryadi

    2014-01-01

    White spot syndrome virus (WSSV) is by far the most devastating shrimp virus. Control measures have lowered the WSSV incidence to various degrees, but the pathogen remains plaguing shrimp culture worldwide. Continuous exposure may cause WSSV to adapt and infect non-crustacean benthic fauna in ponds

  2. The role of white spot syndrome virus (WSSV) VP466 protein in shrimp antiviral phagocytosis.

    Ye, Ting; Zong, Rongrong; Zhang, Xiaobo

    2012-08-01

    Widespread evidence indicates that the structural proteins of virus play very important roles in virus-host interactions. However, the effect of viral proteins on host immunity has not been addressed. Our previous studies revealed that the host shrimp Rab6 (termed as PjRab previously), tropomyosin, β-actin and the white spot syndrome virus (WSSV) envelope protein VP466 formed a complex. In this study, the VP466 protein was shown to be able to bind host Rab6 protein and increase its GTPase activity in vivo and vitro. Thus, VP466 could function as a GTPase-activating protein (GAP) of Rab6. In the VP466-Rab-actin pathway, the increase of the Rab6 activity induced rearrangements of the actin cytoskeleton, resulting in the formation of actin stress fibers which promoted the phagocytosis against virus. Therefore our findings revealed that a viral protein could be employed by host to initiate the host immunity, representing a novel molecular mechanism in the virus-host interaction. Our study would help to better understand the molecular events in immune response against virus infection in invertebrates.

  3. Multiple proteins of White spot syndrome virus involved in recognition of -integrin

    Jing-Yan Zhang; Qing-Hui Liu; Jie Huang

    2014-06-01

    The recognition and attachment of virus to its host cell surface is a critical step for viral infection. Recent research revealed that -integrin was involved in White spot syndrome virus (WSSV) infection. In this study, the interaction of -integrin with structure proteins of WSSV and motifs involved in WSSV infection was examined. The results showed that envelope proteins VP26, VP31, VP37, VP90 and nucleocapsid protein VP136 interacted with LvInt. RGD-, YGL- and LDV-related peptide functioned as motifs of WSSV proteins binding with -integrin. The -integrin ligand of RGDT had better blocking effect compared with that of YGL- and LDV-related peptides. In vivo assay indicated that RGD-, LDV- and YGL-related peptides could partially block WSSV infection. These data collectively indicate that multiple proteins were involved in recognition of -integrin. Identification of proteins in WSSV that are associated with -integrin will assist development of new agents for effective control of the white spot syndrome.

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

    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.

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

    Upasana Ghosh; Somnath Chakraborty; Thangavel Balasubramanian; Punyabrata Das

    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.

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

    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.

  7. White Spot Syndrome Virus infection in Penaeus monodon is facilitated by housekeeping molecules

    Vinayak Biradar; Santosh Narwade; Mandar Paingankar; Deepti Deobagkar

    2013-12-01

    White Spot Syndrome Virus (WSSV) is a major pathogen in shrimp aquaculture, and its rampant spread has resulted in great economic loss. Identification of host cellular proteins interacting with WSSV will help in unravelling the repertoire of host proteins involved in WSSV infection. In this study, we have employed one-dimensional and two-dimension virus overlay protein binding assay (VOPBA) followed by matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) to identify the host proteins of Penaeus monodon that could interact with WSSV. The VOPBA results suggest that WSSV interacted with housekeeping proteins such as heat shock protein 70, ATP synthase subunit , phosphopyruvate hydratase, allergen Pen m 2, glyceraldehyde-3-phosphate dehydrogenase, sarcoplasmic calcium-binding protein, actin and 14-3-3-like protein. Our findings suggest that WSSV exploits an array of housekeeping proteins for its transmission and propagation in P. monodon.

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

    Shashikumar, Anumol; Desai, P V

    2013-03-01

    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 appearance, shrinkage and cell lysis. WSSV infection of cultured cells was confirmed by Nested PCR technique. The incorporation of viral DNA in cultured cells was shown by RAPD profile generated using 10-mer primers. The controls that were not exposed to WSSV did not show cytopathic effects. This work shows the usefulness of proliferating testicular cell culture for studying WSSV infection using molecular tools. Thus, this report gains significance as it opens new vistas for diagnostics and drugs for WSSV.

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

    Liu, Yingjie; Wu, Jinlu; Chen, Hu; Hew, Choy Leong; Yan, Jie

    2010-12-20

    The White Spot Syndrome Virus (WSSV) has a large circular double-stranded DNA genome of around 300kb 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.

  10. The effects of a thermophile metabolite, tryptophol, upon protecting shrimp against white spot syndrome virus.

    Zhu, Fei; Jin, Min

    2015-12-01

    White spot syndrome virus (WSSV) is a shrimp pathogen responsible for significant economic loss in commercial shrimp farms and until now, there has been no effective approach to control this disease. In this study, tryptophol (indole-3-ethanol) was identified as a metabolite involved in bacteriophage-thermophile interactions. The dietary addition of tryptophol reduced the mortality in shrimp Marsupenaeus japonicus when orally challenged with WSSV. Our results revealed that 50 mg/kg tryptophol has a better protective effect in shrimp than 10 or 100 mg/kg tryptophol. WSSV copies in shrimp were reduced significantly (P < 0.01) when supplemented with 50 mg/kg tryptophol, indicating that virus replication was inhibited by tryptophol. Consequently, tryptophol represents an effective antiviral dietary supplement for shrimp, and thus holds significant promise as a novel and efficient therapeutic approach to control WSSV in shrimp aquaculture.

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

    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.

  12. Hijacking of host calreticulin is required for the white spot syndrome virus replication cycle.

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

    2014-07-01

    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 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

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

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

    2013-10-01

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

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

    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.

  15. Optimized PCR assay for detection of white spot syndrome virus (WSSV).

    Nunan, Linda M; Lightner, Donald V

    2011-01-01

    A rapid PCR assay for detection of white spot syndrome virus (WSSV) was developed based on the nested PCR procedure described by Lo et al. (1996) and outlined as the recommended PCR diagnostic assay in the Manual of Diagnostic Tests for Aquatic Animals published by the Office of International Epizootics (OIE, 2009). The optimized procedure incorporated the second step primers used in the nested WSSV PCR. By adjusting the annealing temperature and shortening the cycling times, this modified assay is substantially faster and as sensitive as the recommended OIE protocol. The modified PCR test was compared directly to the two-step nested PCR protocol and a modified nested procedure. The sensitivity of the published assay was determined by template dilutions of semi-purified WSSV virions that had been quantitated using real-time PCR for detection of WSSV. Various isolates were tested using the modified procedure, to ensure that the assay was able to detect WSSV from different geographical locations.

  16. Identification of the immediate-early genes of white spot syndrome virus.

    Li, Fang; Li, Mingyuan; Ke, Wei; Ji, Yongchang; Bian, Xiaofang; Yan, Xiumin

    2009-03-01

    During viral infection, viral immediate-early (IE) genes encode regulatory proteins critical for the viral life cycle. Here we screened white spot syndrome virus (WSSV) IE genes with cycloheximide (CHX)-treated primary culture of crayfish hemocyte and a WSSV genome tiling microarray. Sixteen ORFs, including a known WSSV IE gene (ie1/wsv069), were identified and confirmed by RT-PCR and time course studies. The 16 identified IE proteins contain four proteins (wsv051, wsv069, wsv100, wsv079) with transcription activity, one (wsv083) with Ser/Thr kinase domain and one (wsv249) previously described to function as an ubiquitin E3 ligase. Furthermore, most of the identified WSSV IE genes cluster in a 14 kb genomic region (WSSV China isolate: 36,052 to 50,300 bp). This type of arrangement may facilitate the coordinate control and rapid expression of IE genes.

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

    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.

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

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

    2014-07-01

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

  19. White spot syndrome virus enters crayfish hematopoietic tissue cells via clathrin-mediated endocytosis.

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

    2015-12-01

    White spot syndrome virus (WSSV) is a major pathogen of aquacultured shrimp. However, the mechanism of its entry remains poorly understood. In this study, by analyzing the internalization of WSSV using crayfish hematopoietic tissue (HPT) cells, we showed that WSSV virions were engulfed by cell membrane invaginations sharing the features of clathrin-coated pits and then internalized into coated cytoplasmic vesicles. Further investigation indicated that WSSV internalization was significantly inhibited by chlorpromazine (CPZ) but not genistein. The internalized virions were colocalized with endogenous clathrin as well as transferrin which undergoes clathrin-dependent uptake. Preventing endosome acidification by ammonium chloride (NH4Cl) or chloroquine (CQ) dramatically reduced WSSV entry as well. Moreover, disturbance of dynamin activity or depletion of membrane cholesterol also blocked WSSV uptake. These data indicate that WSSV enters crayfish HPT cells via clathrin-mediated endocytosis in a pH-dependent manner, and membrane cholesterol as well as dynamin is critical for efficient viral entry.

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

    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.

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

    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.

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

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

    2011-07-01

    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.

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

    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.

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

    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 w

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

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

  6. Proteomic analysis of differentially expressed proteins in Fenneropenaeus chinensis hemocytes upon white spot syndrome virus infection.

    Wei Li

    Full Text Available 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.

  7. Transcriptional analysis of the DNA polymerase gene of shrimp white spot syndrome virus.

    Chen, Li-Li; Wang, Han-Ching; Huang, Chiu-Jung; Peng, Shao-En; Chen, Yen-Gu; Lin, Shin-Jen; Chen, Wei-Yu; Dai, Chang-Feng; Yu, Hon-Tsen; Wang, Chung-Hsiung; Lo, Chu-Fang; Kou, Guang-Hsiung

    2002-09-15

    The white spot syndrome virus DNA polymerase (DNA pol) gene (WSSV dnapol) has already been tentatively identified based on the presence of highly conserved motifs, but it shows low overall homology with other DNA pols and is also much larger (2351 amino acid residues vs 913-1244 aa). In the present study we perform a transcriptional analysis of the WSSV dnapol gene using the total RNA isolated from WSSV-infected shrimp at different times after infection. Northern blot analysis with a WSSV dnapol-specific riboprobe found a major transcript of 7.5 kb. 5'-RACE revealed that the major transcription start point is located 27 nucleotides downstream of the TATA box, at the nucleotide residue A within a CAGT motif, one of the initiator (Inr) motifs of arthropods. In a temporal expression analysis using differential RT-PCR, WSSV dnapol transcripts were detected at low levels at 2-4 h.p.i., increased at 6 h.p.i., and remained fairly constant thereafter. This is similar to the previously reported transcription patterns for genes encoding the key enzyme of nucleotide metabolism, ribonucleotide reductase. Phylogenetic analysis showed that the DNA pols from three different WSSV isolates form an extremely tight cluster. In addition, similar to an earlier phylogenetic analysis of WSSV protein kinase, the phylogenetic tree of viral DNA pols further supports the suggestion that WSSV is a distinct virus (likely at the family level) that does not belong to any of the virus families that are currently recognized.

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

    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.

  9. Purification of white spot syndrome virus by iodixanol density gradient centrifugation.

    Dantas-Lima, J J; Corteel, M; Cornelissen, M; Bossier, P; Sorgeloos, P; Nauwynck, H J

    2013-10-01

    Up to now, only a few brief procedures for purifying white spot syndrome virus (WSSV) have been described. They were mainly based on sucrose, NaBr and CsCl density gradient centrifugation. This work describes for the first time the purification of WSSV through iodixanol density gradients, using virus isolated from infected tissues and haemolymph of Penaeus vannamei (Boone). The purification from tissues included a concentration step by centrifugation (2.5 h at 60,000 g) onto a 50% iodixanol cushion and a purification step by centrifugation (3 h at 80,000 g) through a discontinuous iodixanol gradient (phosphate-buffered saline, 5%, 10%, 15% and 20%). The purification from infected haemolymph enclosed a dialysis step with a membrane of 1,000 kDa (18 h) and a purification step through the earlier iodixanol gradient. The gradients were collected in fractions and analysed. The number of particles, infectivity titre (in vivo), total protein and viral protein content were evaluated. The purification from infected tissues gave WSSV suspensions with a very high infectivity and an acceptable purity, while virus purified from haemolymph had a high infectivity and a very high purity. Additionally, it was observed that WSSV has an unusually low buoyant density and that it is very sensitive to high external pressures.

  10. In vivo neutralization assays by monoclonal antibodies against white spot syndrome virus in crayfish (Cambarus proclarkii)

    WANG Yinan; ZHAN Wenbin; XING Jing; JIANG Yousheng

    2008-01-01

    The neutralizing activities of eight monoclonal antibodies (MAbs) against white spot syndrome virus (WSSV) (2D2,2B2,1D2,1DS,1C2,4A1,6A4 and 6B4) were analyzed by in vivo experiments.Gills from WSSV-infected shrimp were homogenized and ten-fold serially diluted by PBS,and then incubated with MAbs (hybridoma culture supernatant),respectively.The mixture of WSSV and MAbs were injected into crayfish (Procambarus clarkii).Mter challenge,the death rates of crayfish were counted to determine the neutralizing activities of MAbs.At the same time,the mixture of myeloma culture supernatant and WSSV or PBS was served as positive or negative control,respectively.The results showed that,at each virus dilution,the mean time to death of the crayfish injected with MAb-treated virus was significantly longer than that in the positive control,though they all showed 100% mortality within 25 d,and meanwhile,few crayfish died in the negative control.Among the eight MAbs,2D2,2B2,1D2 and 1DS,especially the former two,delayed the mortality significantly,and 1C2,4Al and 6A4 delayed the mortality as well but not so efficiently,while MAb 6B4 was efficient only when the virus concentration increased.The results indicated that the anti-WSSV MAbs can neutralize WSSV in different virus dilutions.

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

    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.

  12. METODE DETEKSI CEPAT WHITE SPOT SYNDROME VIRUS (WSSV DAN INFECTIUOS MYONECROSIS VIRUS (IMNV MENGGUNAKAN PORTABEL/MOBILE POLYMERASE CHAIN REACTION

    Isti Koesharyani

    2015-06-01

    Full Text Available Budidaya udang Penaeus monodon dan Litopenaeus vannamei di Indonesia merupakan komoditas primadona untuk ekspor dan merupakan salah satu spesies unggulan dalam program budidaya pada Kementerian Kelautan dan Perikanan. Kendala pada budidaya udang tersebut adalah penyakit yang disebabkan oleh infeksi virus, terutama white spot syndrome virus (WSSV dan infectiuos myonecrosis virus (IMNV. Deteksi secara molekuler yang berbasis DNA merupakan metode deteksi sangat akurat dan tepat. Saat ini sudah ada metode deteksi PCR sederhana yang dapat diaplikasi langsung di lapangan, yaitu dengan menggunakan alat Portable Polymerase Chain Reaction Pockit (iiPCR. Tujuan penelitian ini adalah untuk melakukan deteksi cepat WSSV dan IMNV pada udang P. monodon dan L. vannamei dengan menggunakan portabel-PCR. Aplikasi deteksi ini diujicobakan pada sampel udang yang diawetkan dan sudah diketahui kondisinya. Hasil analisis memperlihatkan hasil yang sesuai dengan hasil analisis sebelumnya dengan menggunakan PCR konvensional. Metode ini relatif lebih sederhana, cepat (hanya memerlukan waktu kurang dari 90 menit dan dapat diketahui hasilnya tanpa adanya proses elektroforesis. Metode deteksi ini sangat membantu pembudidaya dalam monitoring penyakit di lapangan secara on the spot, serta cepat dan tepat.

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

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

    2016-10-01

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

  14. Identification of the Nucleocapsid, Tegument, and Envelope Proteins of the Shrimp White Spot Syndrome Virus Virion

    Tsai, Jyh-Ming; Wang, Han-Ching; Leu, Jiann-Horng; Wang, Andrew H.-J.; Zhuang, Ying; Walker, Peter J.; Kou, Guang-Hsiung; Lo, Chu-Fang

    2006-01-01

    The protein components of the white spot syndrome virus (WSSV) virion have been well established by proteomic methods, and at least 39 structural proteins are currently known. However, several details of the virus structure and assembly remain controversial, including the role of one of the major structural proteins, VP26. In this study, Triton X-100 was used in combination with various concentrations of NaCl to separate intact WSSV virions into distinct fractions such that each fraction contained envelope and tegument proteins, tegument and nucleocapsid proteins, or nucleocapsid proteins only. From the protein profiles and Western blotting results, VP26, VP36A, VP39A, and VP95 were all identified as tegument proteins distinct from the envelope proteins (VP19, VP28, VP31, VP36B, VP38A, VP51B, VP53A) and nucleocapsid proteins (VP664, VP51C, VP60B, VP15). We also found that VP15 dissociated from the nucleocapsid at high salt concentrations, even though DNA was still present. These results were confirmed by CsCl isopycnic centrifugation followed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and liquid chromatography-nanoelectrospray ionization-tandem mass spectrometry, by a trypsin sensitivity assay, and by an immunogold assay. Finally, we propose an assembly process for the WSSV virion. PMID:16501111

  15. Characterization and interactome study of white spot syndrome virus envelope protein VP11.

    Wang-Jing Liu

    Full Text Available 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.

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

    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.

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

    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.

  18. Characterization and interactome study of white spot syndrome virus envelope protein VP11.

    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.

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

    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

  20. Development and application of antibody microarray for white spot syndrome virus detection in shrimp

    XU Xiaoli; SHENG Xiuzhen; ZHAN Wenbin

    2011-01-01

    Detecting white spot syndrome virus (WSSV) in shrimp in high efficiency and veracity is important for disease prevention in aquaculture.Antibody-based microarray is a novel proteomic technology that can meet the requirements.In this study,we developed an antibody microarray for WSSV-detection in a specific and parallel way at multiple samples.First,seven slides each with different modifications were characterized by atomic force microscope,and were compared in the efficiency of immobilizing proteins.Of the seven,3-dimensional structured agarose gel-modified slides were chosen appropriate for the microarray for having higher signal value and superior spot size.A purified rabbit anti-WSSV antibody was arrayed as the capture antibody of the microarray on the agarose gel-modified slides,and then the microarray slides were incubated in the tissue homogenate of sampled shrimp and the antibody-antigen complex was detected by Cy3-conjugated anti-WSSV monoclonal antibody.The results were measured by a laser chipscanner and analyzed with software.To obtain satisfied fluorescence signal intensity,optimal conditions were searched.The detection limit of the antibody microarray for WSSV is 0.62 μg/mL,with a proven long shelf life for 6 months at 4℃ or 8 months at -20℃.Furthermore,concordance between antibody microarray and traditional indirect ELISA reached 100% for WSSV detection.These results suggest that the antibody microarray could be served as an effective tool for diagnostic and epidemiological studies of WSSV.

  1. Screening white spot syndrome virus (WSSV)-resistant molecular markers from Fenneropenaeus chinensis

    Wu, Yingying; Meng, Xianhong; Kong, Jie; Luan, Sheng; Luo, Kun; Wang, Qingyin; Zheng, Yongyun

    2017-02-01

    White spot syndrome virus (WSSV)-resistant molecular markers were screened from the selectively bred new variety `Huanghai No. 2' of Fenneropenaeus chinensis using unlabeled-probe high-resolution melting (HRM) technique. After the artificial infection with WSSV, the first 96 dead shrimps and the last 96 surviving shrimps were collected, representing WSSV-susceptible and -resistant populations, respectively. The genotypes at well-developed 39 single nucleotide polymorphisms (SNPs) loci were obtained. As revealed in the Chi-square test, 3 SNPs, genotype A/A of contig C364-89AT, genotype A/A of C2635-527CA and genotype C/T of contig C12355-592CT, were positively correlated with disease-resistance traits. Other 2 SNPs, genotype G/G of contig C283-145AG and genotype C/C of contig C12355-592CT, were negatively correlated. Moreover, analysis with BlastX program for disease-resistant SNPs indicated that 3 contigs, Contig283, Contig364 and Contig12355, matched to the functional genes of effector caspase of Penaeus monodon, peptide transporter family 1-like protein, and 40S ribosomal protein S2 of Perca flavescens with high sequence similarity. The results will be helpful to provide theoretical and technical supports for molecular marker-assisted selective breeding of F. chinensis.

  2. Genotyping of White Spot Syndrome Virus in Chinese Cultured Shrimp during 1998-1999

    Yan-wei Tan; Zheng-li Shi

    2011-01-01

    Recent studies showed that white spot syndrome virus(WSSV)isolates from different geographic locations share a high genetic similarity except the variable regions in ORF23/24 and ORF14/15,and variable number of tandem repeats(VNTR)within ORF94.In this study,genotyping was performed according to these three variable regions among WSSV isolates collected during 1998/1999 from Southern China.These WSSV isolates contain a deletion of 1168,5657,5898,9316 and 11093 bp,respectively in the variable region ORF23/24compared with WSSV-TW,and a deletion of 4749 or 5622 bp in the variable region ORF14/15 relative to TH-96-II.Four types of repeat units(RUs)(6,8,9 and 13 RUs)in ORF94 were detected in these isolates,with the shortest 6 RUs as the most prevalent type.Our results provide important information for a better understanding of the spatio-temporal transmission mode and the WSSV genetic evolution lineage.

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

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

    2013-11-01

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

  4. Electroless-plated gold films for sensitive surface plasmon resonance detection of white spot syndrome virus.

    Lei, Yun; Chen, Hongyu; Dai, Heping; Zeng, Zhaorui; Lin, Yi; Zhou, Feimeng; Pang, Daiwen

    2008-02-28

    The paper describes the rapid and label-free detection of the white spot syndrome virus (WSSV) using a surface plasmon resonance (SPR) device based on gold films prepared by electroless plating. The plating condition for obtaining films suitable for SPR measurements was optimized. Gold nanoparticles adsorbed on glass slides were characterized by transmission electron microscopy (TEM). Detection of the WSSV was performed through the binding between WSSV in solution and the anti-WSSV single chain variable fragment (scFv antibody) preimmobilized onto the sensor surface. Morphologies of the as-prepared gold films, gold films modified with self-assembled alkanethiol monolayers, and films covered with antibody were examined using an atomic force microscope (AFM). To demonstrate the viability of the method for real sample analysis, WSSV of different concentrations present in a shrimp hemolymph matrix was determined upon optimizing the surface density of the antibody molecules. The SPR device based on the electroless-plated gold films is capable of detecting concentration of WSSV as low as 2.5 ng/mL in 2% shrimp hemolymph, which is one to two orders of magnitude lower than the level measurable by enzyme-linked immunosorbant assays.

  5. ICP35 Is a TREX-Like Protein Identified in White Spot Syndrome Virus

    Phairoh, Panapat; Suthibatpong, Thana; Rattanarojpong, Triwit; Jongruja, Nujarin; Senapin, Saengchan; Choowongkomon, Kiattawee; Khunrae, Pongsak

    2016-01-01

    ICP35 is a non-structural protein from White spot syndrome virus believed to be important in viral replication. Since ICP35 was found to localize in the host nucleus, it has been speculated that the function of ICP35 might be involved in the interaction of DNA. In this study, we overexpressed, purified and characterized ICP35. The thioredoxin-fused ICP35 (thio-ICP35) was strongly expressed in E. coli and be able to form itself into dimers. Investigation of the interaction between ICP35 and DNA revealed that ICP35 can perform DNase activity. Structural model of ICP35 was successfully built on TREX1, suggesting that ICP35 might adopt the folding similar to that of TREX1 protein. Several residues important for dimerization in TREX1 are also conserved in ICP35. Residue Asn126 and Asp132, which are seen to be in close proximity to metal ions in the ICP35 model, were shown through site-directed mutagenesis to be critical for DNase activity. PMID:27348862

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

    Tsai, M F; Lo, C F; van Hulten, M C; Tzeng, H F; Chou, C M; Huang, C J; Wang, C H; Lin, J Y; Vlak, J M; Kou, G H

    2000-11-10

    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, respectively. RNA was isolated from WSSV-infected shrimp at different times after infection and Northern blot analysis with rr1- and rr2-specific riboprobes found major transcripts of 2.8 and 1.4 kb, respectively. 5' RACE showed that the major rr1 transcript started at a position of -84 (C) relative to the ATG translational start, while transcription of the rr2 gene started at nucleotide residue -68 (T). A consensus motif containing the transcriptional start sites for rr1 and rr2 was observed (TCAc/tTC). Northern blotting and RT-PCR showed that the transcription of rr1 and rr2 started 4-6 h after infection and continued for at least 60 h. The rr1 and rr2 genes thus appear to be WSSV "early genes."

  7. Construction of white spot syndrome virus (WSSV) whole genome phage display library

    ZHU Yanbing; YANG Feng

    2007-01-01

    A rebuilt vector pCANTAB 5 EE was obtained by inserting a 34 bp double-stranded oligonucleotide which contained a EcoRV recognition sequence into pCANTAB 5 E. White spot syndrome virus (WSSV) genome DNA was fragmented by sonication to isolate fragments mainly in the range of 0.8 ~2.0 kb, then the fragments were blunt-ended with T4 DNA polymerase and cloned into the EcoRV site of pCANTAB 5 EE. The primary recombinant clone of the library was 3.0 × 105.Colony PCR of random selected recombinants showed that the size of the inserts was 0.12 ~ 1.77 kb. After the whole library recombinant phages infected Escherichia coli HB2151 cells, the extracellular and periplasmic extracts were dropped on PVDF membranes to perform dot blot, using polyclonal mouse anti-VP24 serum,anti-WSV026 serum,anti-WSV063 serum,anti-WSV069 serum,anti-WSV112 serum, anti WSV238 serum,anti-WSV303 serum and anti-VP26 serum as the primary antibody, respectively. The results showed that the display library could express the viral proteins.

  8. White spot syndrome virus VP12 interacts with adenine nucleotide translocase of Litopenaeus vannamei.

    Ma, Fang-fang; Chou, Zhi-guang; Liu, Qing-hui; Guan, Guangkuo; Li, Chen; Huang, Jie

    2014-05-01

    White spot syndrome virus VP12 contains cell attachment motif RGD which is considered to be critical for host cell binding. Until now, the function of this protein remains undefined. In this study, we explored the interaction of VP12 with host cells. A new shrimp protein (adenine nucleotide translocase of Litopenaeus vannamei, LvANT) is selected by far-western overlay assay. Tissue distribution of adenine nucleotide translocase mRNA showed that it was commonly spread in all the tissues detected. Cellular localization of LvANT in shrimp hemocytes showed that it was primarily located in the cytoplasm of hemocytes and colocalized with mitochondria. ELISA and far-western blot assay confirmed that VP12 interacted with LvANT. In vivo neutralization assay showed that anti-LvANT antibody can significantly reduce the mortality of shrimp challenged by WSSV at 48h post-treatment. Our results collectively showed that VP12 is involved in host cell binding via interaction with adenine nucleotide translocase.

  9. Development of an immuno-based colorimetric assay for white spot syndrome virus.

    Loyprasert-Thananimit, Suchera; Saleedang, Akrapon; Deachamag, Panchalika; Waiyapoka, Thanyaporn; Neulplub, Maitee; Chotigeat, Wilaiwan

    2014-01-01

    White spot syndrome virus (WSSV) is a major cause of infectious disease in cultured shrimp. A fast and reliable method for detecting and monitoring the amount of WSSV during farming would be extremely useful. This work describes a sandwich immunoassay that uses anti-GST-VP26, a WSSV-binding protein (WBP), and modified streptavidin magnesphere paramagnetic particles (SMPPs) to develop the technique. The WBP was immobilized on SMPPs and later bound to different copies of WSSV. The binding was detected using anti-GST-VP26 conjugated to alkaline phosphatase. This enzymatic reaction successfully changed the test solution to a concentration-dependent yellow color that was measured at 405 nm. The sensitivity of this method was between 1.6 × 10(4) and 1.6 × 10(7) copies µL(-1) of WSSV. In this study, the color for detection and semiquantitative analysis is easily observed and measured and can lead to the development of a test kit for screening WSSV during shrimp farming.

  10. Hematological Changes m White Spot Syndrome Virus-Infected Shrimp, Fenneropenaeus chinensis (Osbeck)

    FENG Shouming; ZHAN Wenbin; XING Jing; LI Jun; YANG Kai; WANG Jing

    2008-01-01

    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 manifesta- tions 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 dis- integrated 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 bemocytes 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.

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

    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.

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

    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.

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

    Somnath Chakraborty; Upasana Ghosh; Thangavel Balasubramanian; Punyabrata Das

    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: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:Thirty species of marine plants were subjected to Soxhlet extraction using water, 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 1000 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:Nine plant isolates exhibited significant survivability in host. The drug MP07X thus The drug MP07X derived from Rhizophora mucronata is a potent anti-WSSV drug.

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

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

    2016-05-01

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

  15. Litopenaeus vannamei clathrin coat AP17 involved in white spot syndrome virus infection.

    Wang, Xiu-Fang; Liu, Qing-Hui; Wu, Yin; Huang, Jie

    2016-05-01

    White spot syndrome virus (WSSV) is the main pathogen of shrimp culture, and has brought great losses of the shrimp aquaculture industry every year since it has been found. However, the specific mechanism of the virus into the cell is not very clear. Recent research suggests that clathrin-mediated endocytosis is involved in WSSV infection. By sequence analysis, clathrin coat AP17 is an σ subunit of AP-2 complex which is involved in clathrin-mediated endocytosis. To obtain the full-length sequence of Clathrin coat AP17 of Litopenaeus vannamei (LvCCAP17), the rapid amplification of cDNA ends (RACE) was performed to get the sequence of 3'and 5' end and splicing by DNAMAN. The full-length sequence of LvCCAP17 is 842 bp and expected to encoding 142 amino acids, and the amino acid sequence was analyzed by online software. The mRNA expression of LvCCAP17 in different tissues was carried out with quantitative real-time PCR and the LvCCAP17 was detected in all tested tissues of Litopenaeus vannamei. The transcriptional expression level of LvCCAP17 in epithelium and hepatopancreas was significantly up-regulated after WSSV infection. Far-Western blotting and ELISA assay showed that LvCCAP17 interacted with rVP26 and rVP37. Silencing of LvCCAP17 gene by double-strand RNA (dsRNA) interference significantly delay of cumulative mortality rate in WSSV infected shrimp and reduced the expression level of immediate early gene 1(ie1) and vp28. These results indicated that clathrin-meated endocytosis is responsible for WSSV infection.

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

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

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

  17. Cloning of Litopenaeus vannamei CD63 and it's role in white spot syndrome virus infection.

    Guan, Guang-Kuo; Liu, Qing-Hui; Li, Chen; Huang, Jie

    2016-07-01

    White Spot Syndrome Virus (WSSV) is currently the most serious shrimp pathogen, which has brought huge losses to shrimp industry worldwide. CD63 of shrimp belongs to the tetraspanin superfamily, which plays an important role in signal transduction and immune process. In this paper, CD63 cDNA sequence of Litopenaeus vannamei was cloned using RACE method. The amplified sequence is 1472 bp, with its ORF 744 bp, encoding 247 amino acids. Bioinformatics analysis showed that the sequence of LvCD63 has 93% similarity with Penaeus monodon and 92% similarity with Fenneropenaeus chinensis. Real-time PCR analysis showed that the mRNA levels of LvCD63 expressed in the tissues of hemocytes, gill, epithelial tissue, heart, lymphoid, hepatopancreas, stomach, intestines, muscle and nerve. Among these tissues the highest expression level was showed in the tissue of haemolymph, followed by epithelial tissue, hepatopancreas, and nerve. The lowest expression level of LvCD63 was appeared in the muscle tissue. After WSSV challenge, the expression levels of LvCD63 were both up-regulated in the tissues of gill and epithelial. However the expression level of LvCD63 in hepatopancreas was down-regulated. Far-western blot analysis showed that LvCD63 interacts with VP28, and both VP28N and VP28C fragments interact with LvCD63. Flow cytometry analysis showed that LvCD63 was present on the surface of hemocytes and it is required for binding of WSSV virions. Neutral experiments in vivo showed that LvCD63LEL delayed WSSV infection in shrimp.

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

    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.

  19. Shrimp miRNAs regulate innate immune response against white spot syndrome virus infection.

    Kaewkascholkul, Napol; Somboonviwat, Kulwadee; Asakawa, Shuichi; Hirono, Ikuo; Tassanakajon, Anchalee; Somboonwiwat, Kunlaya

    2016-07-01

    MicroRNAs are short noncoding RNAs of RNA interference pathways that regulate gene expression through partial complementary base-pairing to target mRNAs. In this study, miRNAs that are expressed in white spot syndrome virus (WSSV)-infected Penaeus monodon, were identified using next generation sequencing. Forty-six miRNA homologs were identified from WSSV-infected shrimp hemocyte. Stem-loop real-time RT-PCR analysis showed that 11 out of 16 selected miRNAs were differentially expressed upon WSSV infection. Of those, pmo-miR-315 and pmo-miR-750 were highly responsive miRNAs. miRNA target prediction revealed that the miRNAs were targeted at 5'UTR, ORF, and 3'UTR of several immune-related genes such as genes encoding antimicrobial peptides, signaling transduction proteins, heat shock proteins, oxidative stress proteins, proteinases or proteinase inhibitors, proteins in blood clotting system, apoptosis-related proteins, proteins in prophenoloxidase system, pattern recognition proteins and other immune molecules. The highly conserved miRNA homolog, pmo-bantam, was characterized for its function in shrimp. The pmo-bantam was predicted to target the 3'UTR of Kunitz-type serine protease inhibitor (KuSPI). Binding of pmo-bantam to the target sequence of KuSPI gene was analyzed by luciferase reporter assay. Correlation of pmo-bantam and KuSPI expression was observed in lymphoid organ of WSSV-infected shrimp. These results implied that miRNAs might play roles as immune gene regulators in shrimp antiviral response.

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

    Mark P Zwart

    Full Text Available 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 dynamics underlying the process of WSSV genome shrinkage using empirical data and a simple mathematical model. METHODOLOGY/PRINCIPAL FINDINGS: We genotyped new WSSV isolates from five Asian countries, and analyzed this information together with published data. Genome size appears to stabilize over time, and deletion size in the ORF23/24 variable region was significantly related to the time of the first WSSV outbreak in a particular country. Parameter estimates derived from fitting a simple mathematical model of genome shrinkage to the data support a geometric progression (k<1 of the genomic deletions, with k = 0.371 ± 0.150. CONCLUSIONS/SIGNIFICANCE: The data suggest that the rate of genome shrinkage decreases over time before attenuating. Bioassay data provided support for a link between genome size and WSSV fitness in an aquaculture setting. Differences in genomic deletions between geographic WSSV isolates suggest that WSSV spread did not follow a smooth pattern of geographic radiation, suggesting spread of WSSV over long distances by commercial activities. We discuss two hypotheses for genome shrinkage, an adaptive and a neutral one. We argue in favor of the adaptive hypothesis, given that there is support for a link between WSSV genome size and fitness.

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

    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) β-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.

  2. A Novel Detection Platform for Shrimp White Spot Syndrome Virus Using an ICP11-Dependent Immunomagnetic Reduction (IMR) Assay.

    Liu, Bing-Hsien; Lin, Yu-Chen; Ho, Chia-Shin; Yang, Che-Chuan; Chang, Yun-Tsui; Chang, Jui-Feng; Li, Chun-Yuan; Cheng, Cheng-Shun; Huang, Jiun-Yan; Lee, Yen-Fu; Hsu, Ming-Hung; Lin, Feng-Chun; Wang, Hao-Ching; Lo, Chu-Fang; Yang, Shieh-Yueh; Wang, Han-Ching

    2015-01-01

    Shrimp white spot disease (WSD), which is caused by white spot syndrome virus (WSSV), is one of the world's most serious shrimp diseases. Our objective in this study was to use an immunomagnetic reduction (IMR) assay to develop a highly sensitive, automatic WSSV detection platform targeted against ICP11 (the most highly expressed WSSV protein). After characterizing the magnetic reagents (Fe3O4 magnetic nanoparticles coated with anti ICP11), the detection limit for ICP11 protein using IMR was approximately 2 x 10(-3) ng/ml, and the linear dynamic range of the assay was 0.1~1 x 10(6) ng/ml. In assays of ICP11 protein in pleopod protein lysates from healthy and WSSV-infected shrimp, IMR signals were successfully detected from shrimp with low WSSV genome copy numbers. We concluded that this IMR assay targeting ICP11 has potential for detecting the WSSV.

  3. Virus replication cycle of white spot syndrome virus in secondary cell cultures from the lymphoid organ of Litopenaeus vannamei.

    Li, Wenfeng; Desmarets, Lowiese M B; De Gryse, Gaëtan M A; Theuns, Sebastiaan; Van Tuan, Vo; Van Thuong, Khuong; Bossier, Peter; Nauwynck, Hans J

    2015-09-01

    The replication cycle of white spot syndrome virus (WSSV) was investigated in secondary cell cultures from the lymphoid organ of Litopenaeus vannamei. The secondary cells formed a confluent monolayer at 24 h post-reseeding, and this monolayer could be maintained for 10 days with a viability of 90 %. Binding of WSSV to cells reached a maximum (73 ± 3 % of cells and 4.84 ± 0.2 virus particles per virus-binding cell) at 120 min at 4 °C. WSSV entered cells by endocytosis. The co-localization of WSSV and early endosomes was observed starting from 30 min post-inoculation (p.i.). Double indirect immunofluorescence staining showed that all cell-bound WSSV particles entered these cells in the period between 0 and 60 min p.i. and that the uncoating of WSSV occurred in the same period. After 1 h inoculation at 27 °C, the WSSV nucleocapsid protein VP664 and envelope protein VP28 started to be synthesized in the cytoplasm from 1 and 3 h p.i., and were transported into nuclei from 3 and 6 h p.i., respectively. The percentage of cells that were VP664- and VP28-positive in their nuclei peaked (50 ± 4 %) at 12 h p.i. Quantitative PCR showed that WSSV DNA started to be synthesized from 6 h p.i. In vivo titration of the supernatants showed that the progeny WSSV were released from 12 h p.i. and peaked at 18 h p.i. In conclusion, the secondary cell cultures from the lymphoid organ were proven to be ideal for examination of the replication cycle of WSSV.

  4. Role of Marsupenaeus japonicus crustin-like peptide against Vibrio penaeicida and white spot syndrome virus infection.

    Hipolito, Sheryll Grospe; Shitara, Aiko; Kondo, Hidehiro; Hirono, Ikuo

    2014-10-01

    Crustins are important AMP that has been identified in crustaceans. In this study, the role of Marsupenaeus japonicus crustin-like peptide (MjCRS) was examined in vivo by RNA interference (RNAi) using double-stranded RNA (dsRNA). Tissue expression analysis revealed that MjCRS transcripts are expressed in different tissues tested with the highest expression observed in hemocytes. Treatment with double-stranded RNA specific to MjCRS led to a significant reduction of MjCRS transcripts within the hemocytes. When MjCRS was silenced and subsequently infected with Vibrio penaeicida final mortality was significantly higher compared with PBS and dsGFP treated groups. On the other hand, final mortalities of MjCRS silenced and PBS injected groups were not significantly different after infection with white spot virus, however, both are significantly higher compared with dsGFP treated group. V. penaeicida infection significantly decreased MjCRS expression at 3, 6, 12 and 24h followed by significant increase at 48 h post-infection. On the contrary, white spot infection significantly increased MjCRS expression at 6 and 12h and decreased at 48 h post-infection. dsRNA treatment alone decreased total hemocyte counts (THCs) and subsequent V. penaeicida or white spot virus infection further decreased THCs. VP28 gene expression was both similarly increased in PBS injected group and MjCRS silenced group at 24 and 48 h-post infection. Results suggest that MjCRS is involved in antibacterial defense and might not have critical function against viral infection.

  5. MOLECULAR DOCKING ANALYSES OF CYNODON DACTYLON DERIVED PHYTOCHEMICALS AGAINST WHITE SPOT SYNDROME VIRUS (WSSV STRUCTURAL PROTEIN VP26

    Damayanthi Devi.I

    2015-05-01

    Full Text Available White spot disease is a major infectious disease of penaeid shrimps caused by the white spot syndrome virus (WSSV. The viral structural proteins are responsib le for binding virus to the cellular membranes of the host that is being systematically infected. An In silic o attempt was made to identify the potential drug to inhibit the WSSV spread of diseases. For that an effort , was made to deduce the antiviral potentiality of Cynodon dactylon derived phytochemicals with docking tec hnique. To stimulate the structure based drug design the, 3D structure of the VP26 (PDB-ID: 2EDM, a tegument protein thought to be i nvolved in the entry of WSSV nucleocapsid into the host nucleus, is retrieved from PDB datab ase and docking studies are carried out with the sketched phytochemical structures using GOLD software. Among the phytochemicals scr eened, luteolin and apigenin shows the best binding affinity with binding energies of 42.51 and 38.92 K.cal/m ol exhibiting the potential to block VP26 (2EDM protein of WSSV. This study will be helpful in developing novel antiviral drugs from plant sources against aquatic important pathogens.

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

    Shuxia Xue

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

  7. Arabidopsis-derived shrimp viral-binding protein, PmRab7 can protect white spot syndrome virus infection in shrimp.

    Thagun, Chonprakun; Srisala, Jiraporn; Sritunyalucksana, Kallaya; Narangajavana, Jarunya; Sojikul, Punchapat

    2012-09-15

    White spot syndrome virus is currently the leading cause of production losses in the shrimp industry. Penaeus monodon Rab7 protein has been recognized as a viral-binding protein with an efficient protective effect against white spot syndrome infection. Plant-derived recombinant PmRab7 might serve as an alternative source for in-feed vaccination, considering the remarkable abilities of plant expression systems. PmRab7 was introduced into the Arabidopsis thaliana T87 genome. Arabidopsis-derived recombinant PmRab7 showed high binding activity against white spot syndrome virus and a viral envelope, VP28. The growth profile of Arabidopsis suspension culture expressing PmRab7 (ECR21# 35) resembled that of its counterpart. PmRab7 expression in ECR21# 35 reached its maximum level at 5 mg g(-1) dry weight in 12 days, which was higher than those previously reported in Escherichia coli and in Pichia. Co-injection of white spot syndrome virus and Arabidopsis crude extract containing PmRab7 in Litopenaeus vannamei showed an 87% increase in shrimp survival rate at 5 day after injection. In this study, we propose an alternative PmRab7 source with higher production yield, and cheaper culture media costs, that might serve the industry's need for an in-feed supplement against white spot syndrome infection.

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

    Arief Taslihan

    2015-06-01

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

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

    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.

  10. A study of the role of glucose transporter 1 (Glut1) in white spot syndrome virus (WSSV) infection.

    Huang, Huai-Ting; Chan, Hoi-Ling; Shih, Tsai-Yen; Chen, Li-Li

    2015-10-01

    White spot syndrome virus (WSSV) is a large enveloped DNA virus, and it causes a serious disease that has led to severe mortalities of cultured shrimps in many countries. To determine the mechanism of virus entry into the cell and to establish an antiviral strategy, the cell receptor for virus entry and receptor binding protein should be identified. A shrimp cell surface protein, glucose transporter1 (Glut1), was found to interact with WSSV in previous study. In this study, this Glut1 was confirmed to have the ability of transporting glucose, and this gene can also be found in other shrimp species. The interaction between Glut1 and some other WSSV envelope proteins in the infectome structure was verified by far western blot and His pull down assay. In vitro and in vivo neutralization using recombinant partial Glut1 revealed that the large extracellular portion of Glut1 could delay WSSV infection. Also, shrimps which were knocked-down Glut1 gene by treated with dsRNA before WSSV challenge showed decreased mortality. These results indeed provide a direction to develop efficient antiviral strategies or therapeutic methods by using Glut1.

  11. White spot viral disease in penaeid shrimp: a review

    Sangamaheswaran, A.P.; Jeyaseelan, M.J.P.

    2001-01-01

    The white spot viral disease in penaeid shrimp affects the development of the global shrimp industry. This paper reviews the viruses that cause the disease, the transmission of the virus, diagnosis and preventive measures.

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

    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.

  13. Comparison of protein expression profiles of the hepatopancreas in Fenneropenaeus chinensis challenged with heat-inactivated Vibrio anguillarum and white spot syndrome virus.

    Jiang, Hao; Li, Fuhua; Zhang, Jiquan; Zhang, Jinkang; Huang, Bingxin; Yu, Yang; Xiang, Jianhai

    2014-02-01

    Fenneropenaeus chinensis (Chinese shrimp) culture industry, like other Penaeidae culture, has been seriously affected by the shrimp diseases caused by bacteria and virus. To better understand the mechanism of immune response of shrimp to different pathogens, proteome research approach was utilized in this study. Firstly, the soluble hepatopancreas protein samples in adult Chinese shrimp among control, heat-inactivated Vibrio-challenged and white spot syndrome virus-infected groups were separated by 2-DE (pH range, 4-7; sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and pH range, 3-10; tricine-SDS-PAGE). Then the differentially expressed protein spots (≥1.5-fold or ≤0.67-fold averagely of controls) were analyzed by LC-ESI-MS/MS. Using Mascot online database searching algorithm and SEQUEST searching program, 48 and 49 differentially expressed protein spots were successfully identified in response to Vibrio and white spot syndrome virus infection, respectively. Based on these results, we discussed the mechanism of immune response of the shrimp and shed light on the differences between immune response of shrimp toward Vibrio and white spot syndrome virus. This study also set a basis for further analyses of some key genes in immune response of Chinese shrimp.

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

    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.

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

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

    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

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

    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. Thi

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

    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.

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

    Yun-Shiang Chang

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

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

    Zeliha Selamoglu Talas

    2014-07-01

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

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

    Zeliha Selamoglu Talas; Mehmet Fuat Gulhan

    2014-01-01

    Objective: To show hemolymph apoptosis in imported shrimp Litopenaeus vannamei from Hawaii to Iran. Methods: One hundred and eighty shrimps [(7.98±0.54) g] which were collected from a research shrimp farm located in Heleh site in north of Bushehr Province were distributed equally to 6 glass aquariums (50 cmí50 cmí60 cm) as group A in triplicate (imported batch in 2011, without crossing with other generations) with well clean aerated sea water (100 L per aquarium), salinity of 40 ‰ and temperature of 29 °C. Shrimps of group B (produced by crossing the adults of imported batches in 2009 up to 2011) were distributed also among 6 aquariums with the same conditions. Both shrimp groups were injected with concentration of LD50=1í105.4 white spot virus.Results:exposure and reached 100% after 36 h but no mortality was occurred up to 15 d in shrimps of group B. The slide evaluation of hemolymph of group B showed an increasing trend of apoptosis occurrence in all three types of hemolymph cells, hyalinocytes, semi-granulocytes and granulocytes from 24 h to 72 h in contrary to group A that not any apoptosis was observed during the course of the study (15 d). The results showed that in group A, the mortality began approximately 24 h after Conclusions: It is concluded that crossing among the specific pathogen free generations could induce the increasing immunity level through apoptosis to protect them against white spot disease.

  1. Studies on pathogenicity and prevalence of white spot syndrome virus in mud crab, Scylla serrata (Forskal), in Zhejiang Province, China.

    Liu, W; Qian, D; Yan, X J

    2011-02-01

    Mud crab, Scylla serrata (Forskal), is the most commercially important marine crab species in China. In recent years, serious diseases have occurred in major mud crab culture regions in SE China. PCR detection of white spot syndrome virus (WSSV) in diseased mud crabs collected from Zhejiang Province during 2006-2008 showed a prevalence of 34.82%. To study the pathogenicity of WSSV to mud crab, healthy mud crabs were injected intramuscularly with serial 10-fold dilutions of a WSSV inoculum. The cumulative mortalities in groups challenged with 10⁻¹, 10⁻², 10⁻³ and 10⁻⁴ dilutions were 100%, 100%, 66.7% and 38.9% at 10 days post-injection, respectively. All moribund and dead mud crabs except the control group were positive for WSSV by PCR. Based on the viral load of the WSSV inoculum by quantitative real-time PCR, the median lethal dose (LD50) of WSSV in S. serrata was calculated as 1.10 × 10⁶ virus copies/crab, or 7.34 × 10³ virus copies g⁻¹ crab weight. The phenoloxidase, peroxidase and superoxide dismutase activities in haemolymph of WSSV-infected moribund crabs, were significantly lower than the control group, whereas alkaline phosphatase, glutamate-pyruvate transaminase and glutamic-oxaloacetic transaminase were higher than in the control group. WSSV was mainly distributed in gills, subcuticular epithelia, heart, intestine and stomach as shown by immunohistochemical analysis with Mabs against WSSV. The epithelial cells of infected gill showed hypertrophied nuclei with basophilic inclusions. Numerous bacilliform virus particles were observed in nuclei of infected gill cells by transmission electron microscopy. It is concluded that WSSV is a major pathogen of mud crab with high pathogenicity.

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

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

    2017-01-01

    White spot syndrome virus (WSSV) is a major devastating virus in aquaculture industry. A sensitive and selective diagnostic method for WSSV is a pressing need for the early detection and protection of the aquaculture farms. Herein, we first report, a simple electrochemical immunosensor based on methylene blue dye (MB) immobilized graphene oxide modified glassy carbon electrode (GCE/GO@MB) for selective, quick (35 ± 5 mins) and raw sample analysis of WSSV. The immunosensor was prepared by sequential modification of primary antibody, blocking agent (bovine serum album), antigen (as vp28 protein), secondary antibody coupled with horseradish peroxidase (Ab2-HRP) on the GCE/GO@MB. The modified electrode showed a well-defined redox peak at an equilibrium potential (E1/2), −0.4 V vs Ag/AgCl and mediated H2O2 reduction reaction without any false positive result and dissolved oxygen interferences in pH 7 phosphate buffer solution. Under an optimal condition, constructed calibration plot was linear in a range of 1.36 × 10−3 to 1.36 × 107 copies μL−1 of vp28. It is about four orders higher sensitive than that of the values observed with polymerase chain reaction (PCR) and western blot based WSSV detection techniques. Direct electrochemical immunosensing of WSSV in raw tissue samples were successfully demonstrated as a real sample system. PMID:28393929

  3. A putative cell surface receptor for white spot syndrome virus is a member of a transporter superfamily.

    Huai-Ting Huang

    Full Text Available 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 function to transport this sugar. Tissue tropism analysis showed that Glut1 was constitutive and highly expressed in almost all organs. Glut1's localization in shrimp cells was further verified and so was its interaction with Penaeus monodon chitin-binding protein (PmCBP, which was itself identified to interact with an envelope protein complex formed by 11 WSSV envelope proteins. In vitro and in vivo neutralization experiments using synthetic peptide contained WSSV binding domain (WBD showed that the WBD peptide could inhibit WSSV infection in primary cultured hemocytes and delay the mortality in shrimps challenged with WSSV. These findings have important implications for our understanding of WSSV entry.

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

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

    2011-03-01

    Outbreaks of white spot syndrome virus (WSSV) in shrimp culture and the relationship between the virus and virulence are not well understood. Here, we provide evidence showing that 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 markers for the characterization of single- and mixed-genotype infections. The overall prevalence of mixed-genotype WSSV infections was 25.7 %. Non-outbreak ponds had a significantly higher frequency of mixed-genotype infections than outbreak ponds for all VNTR loci, both at the individual shrimp as well as at the pond level. The genetic composition of WSSV populations appears to correlate with the health status of shrimp culture in ponds. The causal relationship between genotypic diversity and disease outbreaks can now be experimentally approached.

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

    Kulkarni, Amod D; Kiron, Viswanath; Rombout, Jan H W M; Brinchmann, Monica F; Fernandes, Jorge M O; Sudheer, Naduvilamuriparampu S; Singh, Bright I S

    2014-07-01

    White spot syndrome virus (WSSV) is a pathogen that causes considerable mortality of the farmed shrimp, Penaeus monodon. Candidate 'vaccines', WSSV envelope protein VP28 and formalin-inactivated WSSV, can provide short-lived protection against the virus. In this study, P. monodon was orally intubated with the aforementioned vaccine candidates, and protein expression in the gut of immunised shrimps was profiled. The alterations in protein profiles in shrimps infected orally with live-WSSV were also examined. Seventeen of the identified proteins in the vaccine and WSSV-intubated shrimps varied significantly compared to those in the control shrimps. These proteins, classified under exoskeletal, cytoskeletal, immune-related, intracellular organelle part, intracellular calcium-binding or energy metabolism, are thought to directly or indirectly affect shrimp's immunity. The changes in the expression levels of crustacyanin, serine proteases, myosin light chain, and ER protein 57 observed in orally vaccinated shrimp may probably be linked to immunoprotective responses. On the other hand, altered expression of proteins linked to exoskeleton, calcium regulation and energy metabolism in WSSV-intubated shrimps is likely to symbolise disturbances in calcium homeostasis and energy metabolism.

  6. Application of a Label-Free Immunosensor for White Spot Syndrome Virus (WSSV) in Shrimp Cultivation Water.

    Waiyapoka, Thanyaporn; Deachamag, Panchalika; Chotigeat, Wilaiwan; Bunsanong, Nittaya; Kanatharana, Proespichaya; Thavarungkul, Panote; Loyprasert-Thananimit, Suchera

    2015-10-01

    White spot syndrome virus (WSSV) is a major pathogen affecting the shrimp industry worldwide. In a preliminary study, WSSV binding protein (WBP) was specifically bound to the VP26 protein of WSSV. Therefore, we have developed the label-free affinity immunosensor using the WBP together with anti-GST-VP26 for quantitative detection of WSSV in shrimp pond water. When the biological molecules were immobilized on a gold electrode to form a self-assembled monolayer, it was then used to detect WSSV using a flow injection system with optimized conditions. Binding between the different copies of WSSV and the immobilized biological molecules was detected by an impedance change (ΔZ″) in real time. The sensitivity of the developed immunosensor was in the linear range of 1.6 × 10(1)-1.6 × 10(6) copies/μl. The system was highly sensitive for the analysis of WSSV as shown by the lack of impedance change when using yellow head virus (YHV). The developed immunosensor could be reused up to 37 times (relative standard deviation (RSD), 3.24 %) with a good reproducibility of residual activity (80-110 %). The immunosensor was simple to operate, reliable, reproducible, and could be applied for the detection and quantification of WSSV in water during shrimp cultivation.

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

    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.

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

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

    2015-05-11

    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.

  9. Studies of the viral binding proteins of shrimp BP53, a receptor of white spot syndrome virus.

    Li, Chen; Gao, Xiao-Xiao; Huang, Jie; Liang, Yan

    2016-02-01

    The specific binding between viral attachment proteins (VAPs) of a virus and its cellular receptors on host cells mediates virus entry into host cells, which triggers subsequent viral infections. Previous studies indicate that F1 ATP synthase β subunit (named BP53), is found on the surface of shrimp cells and involved in white spot syndrome virus (WSSV) infection by functioning as a potential viral receptor. Herein, in a far-western blotting assay, three WSSV proteins with molecular weights of 28 kDa, 37 kDa, and >50 kDa were found to interact with BP53. The 28 kDa and 37 kDa proteins were identified as the envelope protein VP28 and VP37 of WSSV respectively, which could be recognized by the polyclonal antibodies. Enzyme-linked immunosorbent binding assays revealed that VP37 contributed to almost 80% of the binding capability for BP53 compared with the same amount of total WSSV protein. The relationship between BP53 and its complementary interacting protein, VP37, was visualized using a co-localization assay. Bound VP37 on the cell surface co-localized with BP53 and shared a similar subcellular location on the outer surface of shrimp cells. Pearson's correlation coefficients reached to 0.67 ± 0.05 and the Mander's overlap coefficients reached 0.70 ± 0.05, which indicated a strong relationship between the localization of BP53 and bound rVP37. This provides evidence for an interaction between BP53 and VP37 obtained at the molecular and cellular levels, supporting the hypothesis that BP53 serves as a receptor for WSSV by binding to VP37. The identification of the viral binding proteins of shrimp BP53 is helpful for better understanding the pathogenic mechanisms of WSSV to infect shrimp at the cellular level.

  10. Morphology and Molecular Biology of White Spot Syndrome Virus%白斑综合征病毒(White spot syndrome virus)的形态和分子生物学

    王吉桥; 姜玉声; 张建

    2008-01-01

    白斑综合征病毒(white spot syndrome virus,WSSV)自1992年首次发现以来,已成为威胁对虾养殖业的最主要病原.10年中它迅速蔓延至世界各养虾区,造成直接经济损失达700多亿美元.WSSV属线头病毒科Nimaviridae,完整包膜的病毒体(粒子)长210~380 nm,最宽处70~167 nm,病毒体末端具尾状附器(tail-like appendage).WSSV被膜厚6~7 nm,为脂质三层膜结构,由不透明电子层分隔成两层透明层.核衣壳位于膜内,为弹簧体型结构,由直径10 nm的球形蛋白亚基构成.WSSV基因组为环状双链DNA分子,A+T含量占59%,分布均匀,发生分为六个时期.我国WSSV基因组有9.63 kbp的不稳定区,依寄主的不同发生了不同程度的却失,使致毒力发生了变化.

  11. 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

    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. The enhancement of fluorescence was proportional to the initial concentration of the template DNA. A linear relationship was observed between the amount of input plasmid DNA and cycle threshold (CT) values over a range of 1 to 105 copies of the viral genome. To control the variation in sampling and processing among samples, the shrimp β-actin gene was amplified in parallel with the viral DNA. The CT values of IHHNV- and WSV-infected samples were used to determine absolute viral copy numbers from the standard CT curves of these viruses. For each virus and its β-actin control, the specificity of amplification was monitored by using the dissociation curve of the amplified product. Using genomic DNA as a template, SYBR Green PCR was found to be 100- to 2000-fold more sensitive than conventional PCR, depending on the virus, for the samples tested. The results demonstrate that SYBR Green PCR can be used as a rapid and highly sensitive detection and quantification method for shrimp viruses and that it is amenable to high-throughout assay. PMID:11474000

  12. Label free detection of white spot syndrome virus using lead magnesium niobate-lead titanate piezoelectric microcantilever sensors.

    Capobianco, Joseph A; Shih, Wei-Heng; Leu, Jiann-Horng; Lo, Grace Chu-Fang; Shih, Wan Y

    2010-11-15

    We have investigated rapid, label free detection of white spot syndrome virus (WSSV) using the first longitudinal extension resonance peak of five lead-magnesium niobate-lead titanate (PMN-PT) piezoelectric microcantilever sensors (PEMS) 1050-700 μm long and 850-485 μm wide constructed from 8 μm thick PMN-PT freestanding films. The PMN-PT PEMS were encapsulated with a 3-mercaptopropyltrimethoxysilane (MPS) insulation layer and further coated with anti-VP28 and anti-VP664 antibodies to target the WSSV virions and nucleocapsids, respectively. By inserting the antibody coated PEMS in a flowing virion or nucleocapsid suspension, label free detection of the virions and nucleocapsids were respectively achieved by monitoring the PEMS resonance frequency shift. We showed that positive label free detection of both the virion and the nucleocapsid could be achieved at a concentration of 100virions(nucleocapsids)/ml or 10 virions(nucleocapsids)/100 μl, comparable to the detection sensitivity of polymerase chain reaction (PCR). However, in contrast to PCR, PEMS detection was label free, in situ and rapid (less than 30 min), potentially requiring minimal or no sample preparation.

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

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

    2017-03-28

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

  14. Characterization and Diagnostic Use of a Monoclonal Antibody for VP28 Envelope Protein of White Spot Syndrome Virus

    Chong-lin Hou; Yu Cao; Rong-hui Xie; Yi-zhen Wang; Hua-hua Du

    2011-01-01

    The gene encoding the VP28 envelope protein of White spot syndrome virus (WSSV)was cloned into expression vector pET-30a and transformed into the Escherichia coli strain BL21.After induction,the recombinant VP28 (rVP28) protein was purified and then used to immunize Balb/c mice for monoclonal antibody (MAb) production.It was observed by immuno-electron microscopy the MAbs specific to rVP28 could recognize native VP28 target epitopes of WSSV and dot-blot analysis was used to detect natural WSSV infection.Competitive PCR showed that the viral level was approximately 104 copies/mg tissue in the dilution of gill homogenate of WSSV-infected crayfish at the detection limit of dot-blot assay.Our results suggest that dot-blot analysis with anti-rVP28 MAb could rapidly and sensitively detect WSSV at the early stages of WSSV infection.

  15. In vitro white spot syndrome virus (WSSV) replication in explants of the heart of freshwater crab, Paratelphusa hydrodomous.

    Nathiga Nambi, K S; Abdul Majeed, S; Sundar Raj, N; Taju, G; Madan, N; Vimal, S; Sahul Hameed, A S

    2012-08-01

    Explants from different organs of freshwater crab, Paratelphusa hydrodomous were prepared to establish an in vitro system for replication of white spot syndrome virus (WSSV) of shrimp. Heart explants were maintained for 53 days without any morphological changes in EX-CELL™ 405 medium with and without serum whereas the explants of eye muscle, gill, shell membrane and appendage muscle died within 15 days of culture period. The heart explants on different days of culture were exposed to WSSV for 10 days to study the viral replication. The infection of WSSV in explants of the heart was confirmed by PCR, RT-PCR, Western blot, histology, immunohistochemistry, bioassay and transmission electron microscopy. The WSSV was quantified by real-time PCR and indirect ELISA. The WSSV inoculum prepared from the heart explants of crab caused significant mortality in Penaeus monodon in challenge experiments and the results indicate that the WSSV which replicated in the heart explants of freshwater crab maintains its infectivity as in marine shrimp. The results indicate that the heart explants of P. hydrodomous would be a good alternative to whole animals for production of WSSV.

  16. Envelope Proteins of White Spot Syndrome Virus (WSSV Interact with Litopenaeus vannamei Peritrophin-Like Protein (LvPT.

    Shijun Xie

    Full Text Available White spot syndrome virus (WSSV is a major pathogen in shrimp cultures. The interactions between viral proteins and their receptors on the surface of cells in a frontier target tissue are crucial for triggering an infection. In this study, a yeast two-hybrid (Y2H library was constructed using cDNA obtained from the stomach and gut of Litopenaeus vannamei, to ascertain the role of envelope proteins in WSSV infection. For this purpose, VP37 was used as the bait in the Y2H library screening. Forty positive clones were detected after screening. The positive clones were analyzed and discriminated, and two clones belonging to the peritrophin family were subsequently confirmed as genuine positive clones. Sequence analysis revealed that both clones could be considered as the same gene, LV-peritrophin (LvPT. Co-immunoprecipitation confirmed the interaction between LvPT and VP37. Further studies in the Y2H system revealed that LvPT could also interact with other WSSV envelope proteins such as VP32, VP38A, VP39B, and VP41A. The distribution of LvPT in tissues revealed that LvPT was mainly expressed in the stomach than in other tissues. In addition, LvPT was found to be a secretory protein, and its chitin-binding ability was also confirmed.

  17. A novel C-type lectin from the shrimp Litopenaeus vannamei possesses anti-white spot syndrome virus activity.

    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

    2009-01-01

    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 (Glu(99)-Pro(100)-Asn(101)) motif that has a predicted ligand binding specificity for mannose. Reverse transcription-PCR analysis revealed that LvCTL1 mRNA was specifically expressed in the hepatopancreas of L. vannamei. Recombinant LvCTL1 (rLvCTL1) had hemagglutinating activity and ligand binding specificity for mannose and glucose. rLvCTL1 also had a strong affinity for WSSV and interacted with several envelope proteins of WSSV. Furthermore, we showed that the binding of rLvCTL1 to WSSV could protect shrimps from viral infection and prolong the survival of shrimps against WSSV infection. Our results suggest that LvCTL1 is a mannose-binding C-type lectin that binds to envelope proteins of WSSV to exert its antiviral activity. To our knowledge, this is the first report of a shrimp C-type lectin that has direct anti-WSSV activity.

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

    Deborah Gnana Selvam

    2012-01-01

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

  19. Envelope Proteins of White Spot Syndrome Virus (WSSV) Interact with Litopenaeus vannamei Peritrophin-Like Protein (LvPT).

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

    2015-01-01

    White spot syndrome virus (WSSV) is a major pathogen in shrimp cultures. The interactions between viral proteins and their receptors on the surface of cells in a frontier target tissue are crucial for triggering an infection. In this study, a yeast two-hybrid (Y2H) library was constructed using cDNA obtained from the stomach and gut of Litopenaeus vannamei, to ascertain the role of envelope proteins in WSSV infection. For this purpose, VP37 was used as the bait in the Y2H library screening. Forty positive clones were detected after screening. The positive clones were analyzed and discriminated, and two clones belonging to the peritrophin family were subsequently confirmed as genuine positive clones. Sequence analysis revealed that both clones could be considered as the same gene, LV-peritrophin (LvPT). Co-immunoprecipitation confirmed the interaction between LvPT and VP37. Further studies in the Y2H system revealed that LvPT could also interact with other WSSV envelope proteins such as VP32, VP38A, VP39B, and VP41A. The distribution of LvPT in tissues revealed that LvPT was mainly expressed in the stomach than in other tissues. In addition, LvPT was found to be a secretory protein, and its chitin-binding ability was also confirmed.

  20. Identification and Characterization of Nuclear Localization Signals within the Nucleocapsid Protein VP15 of White Spot Syndrome Virus

    Li-juan LI; Hua-jun ZHANG; Cong ZHANG; Zheng-li SHI

    2009-01-01

    The nucleocapsid protein VP15 of white spot syndrome virus (WSSV) is a basic DNA-binding protein. Three canonical bipartite nuclear localization signals (NLSs), called NLS1 (aa 11-27), NLS2 (aa 33-49) and NLS3 (44-60), have been detected in this protein, using the ScanProsite computer program. To determine the nuclear localization sequence of VP15, the full-length open reading frame, or the sequence of one of the three NLSs, was fused to the green fluorescent protein (GFP) gene, and transiently expressed in insect Sf9 cells. Transfection with full-length VP15 resulted in GFP fluorescence being distributed exclusively in the nucleus. NLS 1 alone could also direct GFP to the nucleus, but less efficiently. Neither of the other two NLSs (NLS2 and 3) was functional when expressed alone, but exhibited similar activity to NLS1 when they were expressed as a fusion peptide. Furthermore, a mutated VP15, in which the two basic amino acids (11RR12) of NLSI were changed to two alanines (11AA12), caused GFP to be localized only in the cytoplasm of Sf9 cells. These results demonstrated that VP15, as a nuclear localization protein, needs cooperation between its three NLSs, and that the two residues (11RR12) of NLS1 play a key role in transporting the protein to the nucleus.

  1. Graphene oxide based fluorescence resonance energy transfer and loop-mediated isothermal amplification for white spot syndrome virus detection.

    Waiwijit, U; Phokaratkul, D; Kampeera, J; Lomas, T; Wisitsoraat, A; Kiatpathomchai, W; Tuantranont, A

    2015-10-20

    Graphene oxide (GO) is attractived for biological or medical applications due to its unique electrical, physical, optical and biological properties. In particular, GO can adsorb DNA via π-π stacking or non-covalent interactions, leading to fluorescence quenching phenomenon applicable for bio-molecular detection. In this work, a new method for white spot syndrome virus (WSSV)-DNA detection is developed based on loop-mediated isothermal amplification (LAMP) combined with fluorescence resonance energy transfer (FRET) between GO and fluorescein isothiocyanate-labeled probe (FITC-probe). The fluorescence quenching efficiency of FITC-probe was found to increase with increasing GO concentration and reached 98.7% at a GO concentration of 50 μg/ml. The fluorescence intensity of FITC-probe was recovered after hybridization with WSSV LAMP product with an optimal hybridization time of 10 min and increased accordingly with increasing amount of LAMP products. The detection limit was estimated to be as low as 10 copies of WSSV plasmid DNA or 0.6 fg of the total DNA extracted from shrimp infected with WSSV. In addition, no cross reaction was observed with other common shrimp viral pathogens. Therefore, the GO-FRET-LAMP technique is promising for fast, sensitive and specific detection of DNAs.

  2. Relationship between white spot syndrome virus (WSSV) loads and characterizations of water quality in Litopenaeus vannamei culture ponds during the tropical storm

    Zhang, J. S.; Li, Z. J.; Wen, G. L.; Wang, Y. L.; Luo, L.; Zhang, H. J.; Dong, H. B.

    2016-01-01

    An in-situ experiment was conducted to investigate the effect of tropical storm on the white spot syndrome virus (WSSV) loads in Litopenaeus vannamei rearing ponds. White spot syndrome virus loads, heterotrophic bacteria, Vibrio and water quality (including temperature, dissolved oxygen (DO), salinity, pH, NH4-N, and NO2-N) were continually monitored through one tropical storm. The WSSV loads decreased when tropical storm made landfall, and substantially increased when typhoon passed. The variation of WSSV loads was correlated with DO, temperature, heterotrophic bacteria count, and ammonia-N concentrations. These results suggested that maintaining high level DO and promoting heterotrophic bacteria growth in the shrimp ponds might prevent the diseases’ outbreak after the landfall of tropical storm. PMID:27822254

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

    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 μL(-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.

  4. White spot syndrome virus induces metabolic changes resembling the warburg effect in shrimp hemocytes in the early stage of infection.

    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

    2011-12-01

    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 WSSV-infected shrimp, and the key enzyme of the pentose phosphate pathway, glucose-6-phosphate dehydrogenase (G6PDH), showed increased activity. We also found that at 12 hpi there was no alteration in the ADP/ATP ratio and that oxidative stress was lower than that in uninfected controls. All of these results are characteristic of the Warburg effect as it is present in mammals. There was also a significant decrease in triglyceride concentration starting at 12 hpi. At the late stage of the infection cycle (24 hpi), hemocytes of WSSV-infected shrimp showed several changes associated with cell death. These included the induction of mitochondrial membrane permeabilization (MMP), increased oxidative stress, decreased glucose consumption, and disrupted energy production. A previous study showed that WSSV infection led to upregulation of the voltage-dependent anion channel (VDAC), which is known to be involved in both the Warburg effect and MMP. Here we show that double-stranded RNA (dsRNA) silencing of the VDAC reduces WSSV-induced mortality and virion copy number. For these results, we hypothesize a model depicting the metabolic changes in host cells at the early and late stages of WSSV infection.

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

    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.

  6. White spot syndrome virus isolates of tiger shrimp Penaeus monodon (Fabricious) in India are similar to exotic isolates as revealed by polymerase chain reaction and electron microscopy.

    Mishra, S S; Shekhar, M S

    2005-07-01

    Microbiological analysis of samples collected from cases of white spot disease outbreaks in cultured shrimp in different farms located in three regions along East Coast of India viz. Chidambram (Tamil Nadu), Nellore (Andhra Pradesh) and Balasore (Orissa), revealed presence of Vibrio alginolyticus, Vibrio parahaemolyticus, and Aeromonas spp. but experimental infection trials in Penaeus monodon with these isolates did not induce any acute mortality or formation of white spots on carapace. Infection trials using filtered tissue extracts by oral and injection method induced mortality in healthy P. monodon with all samples and 100% mortality was noted by the end of 7 day post-inoculation. Histopathological analysis demonstrated degenerated cells characterized by hypertrophied nuclei in gills, hepatopancreas and lymphoid organ with presence of intranuclear basophilic or eosino-basophilic bodies in tubular cells and intercellular spaces. Analysis of samples using 3 different primer sets as used by other for detection of white spot syndrome virus (WSSV) generated 643, 1447 and 520bp amplified DNA products in all samples except in one instance. Variable size virions with mean size in the range of 110 x 320 +/- 20 nm were observed under electron microscope. It could be concluded that the viral isolates in India involved with white spot syndrome in cultured shrimp are similar to RV-PJ and SEMBV in Japan, WSBV in Taiwan and WSSV in Malaysia, Indonesia, Thailand, China and Japan.

  7. Dietary administration of a Gracilaria tenuistipitata extract enhances the immune response and resistance against Vibrio alginolyticus and white spot syndrome virus in the white shrimp Litopenaeus vannamei.

    Sirirustananun, Nuttarin; Chen, Jiann-Chu; Lin, Yong-Chin; Yeh, Su-Tuen; Liou, Chyng-Hwa; Chen, Li-Li; Sim, Su Sing; Chiew, Siau Li

    2011-12-01

    The haemogram, phenoloxidase (PO) activity, respiratory bursts (RBs), superoxide dismutase (SOD) activity, glutathione peroxidase (GPx) activity, lysozyme activity, and the mitotic index of haematopoietic tissue (HPT) were examined after the white shrimp Litopenaeus vannamei had been fed diets containing the hot-water extract of Gracilaria tenuistipitata at 0 (control), 0.5, 1.0, and 2.0 g kg(-1) for 7-35 days. Results indicated that these parameters directly increased with the amount of extract and time, but slightly decreased after 35 days. RBs, SOD activity, and GPx activity reached the highest levels after 14 days, whereas PO and lysozyme activities reached the highest levels after 28 days. In a separate experiment, white shrimp L. vannamei, which had been fed diets containing the extract for 14 days, were challenged with Vibrio alginolyticus at 2 × 10(6) cfu shrimp(-1) and white spot syndrome virus (WSSV) at 1 × 10(3) copies shrimp(-1), and then placed in seawater. The survival rate of shrimp fed the extract-containing diets was significantly higher than that of shrimp fed the control diet at 72-144 h post-challenge. We concluded that dietary administration of the G. tenuistipitata extract at ≤1.0 g kg(-1) could enhance the innate immunity within 14 days as evidenced by the increases in immune parameters and mitotic index of HPT in shrimp and their enhanced resistance against V. alginolyticus and WSSV infections. Shrimp fed the extract-containing diets showed a higher and continuous increase in the humoral response indicating its persistent role in innate immunity.

  8. White spot syndrome virus (WSSV) in Litopenaeus vannamei captured from the Gulf of California near an area of extensive aquaculture activity.

    Mijangos-Alquisires, Z; Quintero-Arredondo, N; Castro-Longoria, R; Grijalva-Chon, J M; Ramos-Paredes, J

    2006-07-11

    For the shrimp farming industry of Mexico, disease outbreaks caused by white spot syndrome virus (WSSV) are relatively recent. Efforts to control the virus are assisted by monitoring for its prevalence in aquaculture systems, but few attempts have been made to search for it in carriers from coastal waters. To search for WSSV carriers in the Gulf of California, we made surveys off the coast of Sinaloa, Mexico, in March 2001, November 2001, and September 2003 using polymerase chain reaction (PCR) assays and histopathology. WSSV-positive shrimp were detected only in November 2001, after hurricane Julliete. This suggested possible dispersal of WSSV to the marine environment from infected shrimp farms.

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

    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.

  10. Injected phage-displayed-VP28 vaccine reduces shrimp Litopenaeus vannamei mortality by white spot syndrome virus infection.

    Solís-Lucero, G; Manoutcharian, K; Hernández-López, J; Ascencio, F

    2016-08-01

    White spot syndrome virus (WSSV) is the most important viral pathogen for the global shrimp industry causing mass mortalities with huge economic losses. Recombinant phages are capable of expressing foreign peptides on viral coat surface and act as antigenic peptide carriers bearing a phage-displayed vaccine. In this study, the full-length VP28 protein of WSSV, widely known as potential vaccine against infection in shrimp, was successfully cloned and expressed on M13 filamentous phage. The functionality and efficacy of this vaccine immunogen was demonstrated through immunoassay and in vivo challenge studies. In ELISA assay phage-displayed VP28 was bind to Litopenaeus vannamei immobilized hemocyte in contrast to wild-type M13 phage. Shrimps were injected with 2 × 10(10) cfu animal(-1) single dose of VP28-M13 and M13 once and 48 h later intramuscularly challenged with WSSV to test the efficacy of the vaccine against the infection. All dead challenged shrimps were PCR WSSV-positive. The accumulative mortality of the vaccinated and challenged shrimp groups was significantly lower (36.67%) than the unvaccinated group (66.67%). Individual phenoloxidase and superoxide dismutase activity was assayed on 8 and 48 h post-vaccination. No significant difference was found in those immunological parameters among groups at any sampled time evaluated. For the first time, phage display technology was used to express a recombinant vaccine for shrimp. The highest percentage of relative survival in vaccinated shrimp (RPS = 44.99%) suggest that the recombinant phage can be used successfully to display and deliver VP28 for farmed marine crustaceans.

  11. Susceptibility of juvenile Macrobrachium rosenbergii to different doses of high and low virulence strains of white spot syndrome virus (WSSV).

    Corteel, Mathias; Dantas-Lima, João J; Tuan, Vo Van; Thuong, Khuong Van; Wille, Mathieu; Alday-Sanz, Victoria; Pensaert, Maurice B; Sorgeloos, Patrick; Nauwynck, Hans J

    2012-09-12

    As some literature on the susceptibility of different life stages of Macrobrachium rosenbergii to white spot syndrome virus (WSSV) is conflicting, the pathogenesis, infectivity and pathogenicity of 2 WSSV strains (Thai-1 and Viet) were investigated here in juveniles using conditions standardized for Penaeus vannamei. As with P. vannamei, juvenile M. rosenbergii (2 to 5 g) injected with a low dose of WSSV-Thai-1 or a high dose of WSSV-Viet developed comparable clinical pathology and numbers of infected cells within 1 to 2 d post-infection. In contrast, a low dose of WSSV-Viet capable of causing mortality in P. vannamei resulted in no detectable infection in M. rosenbergii. Mean prawn infectious dose 50% endpoints (PID₅₀ ml⁻¹) determined in M. rosenbergii were in the order of 100-fold higher for WSSV-Thai-1 (105.3 ± 0.4 PID₅₀ ml⁻¹) than for WSSV-Viet (103.2 ± 0.2 PID₅₀ ml⁻¹), with each of these being about 20-fold and 400-fold lower, respectively, than found previously in P. vannamei. The median lethal dose (LD₅₀ ml⁻¹) determined in M. rosenbergii was also far higher (~1000-fold) for WSSV-Thai-1 (105.4 ± 0.4 LD₅₀ ml⁻¹) than for WSSV-Viet (102.3 ± 0.3 LD₅₀ ml⁻¹). Based on these data, it is clear that juvenile M. rosenbergii are susceptible to WSSV infection, disease and mortality. In comparison to P. vannamei, however, juvenile M. rosenbergii appear more capable of resisting infection and disease, particularly in the case of a WSSV strain with lower apparent virulence.

  12. A gC1qR prevents white spot syndrome virus replication in the freshwater crayfish Pacifastacus leniusculus.

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

    2010-10-01

    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 the Hpt cell culture and live crayfish enhanced the WSSV replication. In addition, by making a recombinant PlgC1qR protein we could show that if this recombinant protein was injected in a crayfish, Pacifastacus leniusculus, followed by injection of WSSV, this significantly reduced viral replication in vivo. Furthermore, if the recombinant PlgC1qR was incubated with Hpt cells and then WSSV was added, this also reduced viral replication. These experiments clearly demonstrate that recombinant PlgC1qR reduce WSSV replication both in vivo and in vitro. The results from a far-Western overlay and glutathione S-transferase pull-down assays showed that PlgC1qR could bind to VP15, VP26, and VP28. Altogether, these results demonstrate a role for PlgC1qR in antiviral activity against WSSV.

  13. Prohibitin Interacts with envelope proteins of white spot syndrome virus and prevents infection in the red swamp crayfish, Procambarus clarkii.

    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

    2013-12-01

    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.

  14. An immersion of Gracilaria tenuistipitata extract improves the immunity and survival of white shrimp Litopenaeus vannamei challenged with white spot syndrome virus.

    Lin, Yong-Chin; Yeh, Su-Tuen; Li, Chang-Che; Chen, Li-Li; Cheng, Ann-Chang; Chen, Jiann-Chu

    2011-12-01

    The innate immunity and resistance against white spot syndrome virus (WSSV) in white shrimp Litopenaeus vannamei which received the Gracilaria tenuistipitata extract were examined. Shrimp immersed in seawater containing the extract at 0 (control), 400 and 600 mg L(-1) for 3 h were challenged with WSSV at 2 × 10(4) copies shrimp(-1). Shrimp not exposed to the extract and not received WSSV challenge served as unchallenged control. The survival rate of shrimp immersed in 400 mg L(-1) or 600 mg L(-1) extract was significantly higher than that of challenged control shrimp over 24-120 h. The haemocyte count, phenoloxidase activity, respiratory burst, superoxide dismutase activity, and lysozyme activity of shrimp immersed in 600 mg L(-1) extract were significantly higher than those of unchallenged control shrimp at 6, 6, 6, 6, and 6-24 h post-challenge. In another experiment, shrimp which had received 3 h immersion of 0, 400, 600 mg L(-1) extract were challenged with WSSV. The shrimp were then received a booster (3 h immersion in the same dose of the extract), and the immune parameters were examined at 12-120 h post-challenge. The immune parameters of shrimp immersed in 600 mg L(-1) extract, and then received a booster at 9, 21, and 45 h were significantly higher than those of unchallenged control shrimp at 12-48 h post-challenge. In conclusion, shrimp which had received the extract exhibited protection against WSSV as evidenced by the higher survival rate and higher values of immune parameters. Shrimp which had received the extract and infected by WSSV showed improved immunity when they received a booster at 9, 21, and 45 h post-WSSV challenge. The extract treatment caused less decrease in PO activity, and showed better performance of lysozyme activity and antioxidant response in WSSV-infected shrimp.

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

    Zhang, Xiaojing; Song, Xiaoling; Huang, Jie

    2016-11-01

    White spot syndrome virus (WSSV) is an important viral pathogen that infects farmed penaeid shrimp, and the threat of Vibrio parahaemolyticus infection to shrimp farming has become increasingly severe. Viral and bacterial cross or superimposed infections may induce higher shrimp mortality. We used a feeding method to infect Litopenaeus vannamei with WSSV and then injected a low dose of V. parahaemolyticus (WSSV+Vp), or we first infected L. vannamei with a low-dose injection of V. parahaemolyticus and then fed the shrimp WSSV to achieve viral infection (Vp+WSSV). The eff ect of V. parahaemolyticus and WSSV co-infection on survival of L. vannamei was evaluated by comparing cumulative mortality rates between experimental and control groups. We also spread L. vannamei hemolymph on thiosulfate citrate bile salt sucrose agar plates to determine the number of Vibrio, and the WSSV copy number in L. vannamei gills was determined using an absolute quantitative polymerase chain reaction (PCR) method. LvMyD88 and Lvakt gene expression levels were detected in gills of L. vannamei by real-time PCR to determine the cause of the diff erent mortality rates. Our results show that (1) the cumulative mortality rate of L. vannamei in the WSSV+Vp group reached 100% on day 10 after WSSV infection, whereas the cumulative mortality rate of L. vannamei in the Vp+WSSV group and the WSSV-alone control group approached 100% on days 11 and 13 of infection; (2) the number of Vibrio in the L. vannamei group infected with V. parahaemolyticus alone declined gradually, whereas the other groups showed significant increases in the numbers of Vibrio ( P<0.05); (3) the WSSV copy numbers in the gills of the WSSV+Vp, Vp+WSSV, and the WSSV-alone groups increased from 105 to 107 /mg tissue 72, 96, and 144 h after infection, respectively. These results suggest that V. parahaemolyticus infection accelerated proliferation of WSSV in L. vannamei and vice versa. The combined accelerated proliferation of both V

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

    Vorawit Ananphongmanee

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

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

    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 α-agglutinin. DNA fragments, harboring fused-gene expression cassettes under control of an alcohol oxidase I (AOX1) promoter were constructed and used to transform the yeast cells. Immunofluorescence microscopy with antibodies specific to both proteins demonstrated that mutated PmRab7 (mPmRab7) and partial VP28 (pVP28) were localized on the cell surfaces of the respective clones, and fluorescence intensity for each was significantly higher than that of control cells by flow cytometry. Enzyme-linked immunosorbant assay (ELISA) using cells displaying mPmRab7 or pVP28 revealed that the binding of specific antibodies for each was dose-dependent, and could be saturated. In addition, the binding of mPmRab7-expressing cells with free VP28, and vice versa was dose dependent. Binding between the two surface-expressed proteins was confirmed by an assay showing agglutination between cells expressing complementary mPmRab7 and pVP28. In summary, our genetically engineered P. pastoris can display biologically active mPmRab7 and pVP28 and is now ready for evaluation of efficacy in protecting shrimp against WSSV by oral

  18. Lymphoid organ cell culture system from Penaeus monodon (Fabricius) as a platform for white spot syndrome virus and shrimp immune-related gene expression.

    Jose, S; Jayesh, P; Sudheer, N S; Poulose, G; Mohandas, A; Philip, R; Singh, I S Bright

    2012-05-01

    Shrimp cell lines are yet to be reported and this restricts the prospects of investigating the associated viral pathogens, especially white spot syndrome virus (WSSV). In this context, development of primary cell cultures from lymphoid organs was standardized. Poly-l-lysine-coated culture vessels enhanced growth of lymphoid cells, while the application of vertebrate growth factors did not, except insulin-like growth factor-1 (IGF-1). Susceptibility of the lymphoid cells to WSSV was confirmed by immunofluoresence assay using monoclonal antibody against the 28 kDa envelope protein of WSSV. Expression of viral and immune-related genes in WSSV-infected lymphoid cultures could be demonstrated by RT-PCR. This emphasizes the utility of lymphoid primary cell culture as a platform for research in virus-cell interaction, virus morphogenesis, up and downregulation of shrimp immune-related genes, and also for the discovery of novel drugs to combat WSSV in shrimp culture.

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

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

    having sensitivity equal to that of 1-step PCR for detection of WSSV. Here we report the evaluation and comparison of the MAb-based immunodot test to PCR as a screening tool for WSSV in Penaeus monodon grow-out farms. © Inter-Research 2008 · www....int-res.com*Corresponding author. Email: kalkulishankar@gmail.com NOTE Evaluation of an immunodot test to manage white spot syndrome virus (WSSV) during cultivation of the giant tiger shrimp Penaeus monodon Rajreddy Patil 1 , K. J. Palaksha 1 , T. M. Anil 1 , Guruchannabasavanna 1...

  20. REVIEW ON SHRIMP WHITE SPOT SYNDROME VIRUS (WSSV) HOSTS%对虾白斑综合征病毒(WSSV)宿主研究进展

    闫冬春

    2007-01-01

    对虾白斑综合征病毒(white spot syndrome virus,WSSV)自1992年暴发以来,已在世界范围内传播开来,每年给对虾养殖业造成巨大经济损失,至今仍未得到完全控制.本文根据1995~2005年的有关文献资料,综述了对虾白斑综合征病毒宿主方面的研究进展.

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

    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.

  2. Haemocyanin content of shrimp (Fenneropenaeus chinensis) associated with white spot syndrome virus and Vibrio harveyi infection process.

    Chang, Yanhong; Xing, Jing; Tang, Xiaoqian; Sheng, Xiuzhen; Zhan, Wenbin

    2016-01-01

    Haemocyanin (Hc) is frequently reported to vary significantly by physiological status and environmental stress in Crustaceans. In this paper, the shrimp Fenneropenaeus chinensis was infected with different concentrations of white spot syndrome virus (WSSV) and Vibrio harveyi. Then, the variation of Hc and total protein content of the haemolymph (TPCH) were investigated using the established double antibody sandwich enzyme linked immunosorbent assay (DAS-ELISA) and Coomassie brilliant blue method, respectively. The results showed that the Hc content peaked at 12 h post-infection (PI) in the 10(-2), 10(-4) and 10(-6) viral supernatant (VS) groups, and the maximum was 93.03 ± 2.55 mg ml(-1), 77.57 ± 6.02 mg ml(-1) and 70.25 ± 3.96 mg ml(-1), respectively. TPCH reached the maximum of 108.18 ± 1.36 mg ml(-1) and 103.49 ± 1.33 mg ml(-1) at 12 h PI in the 10(-2) and 10(-4) VS groups, respectively. The maximum was 96.94 ± 1.06 mg ml(-1) at 24 h PI in the 10(-6) VS group. In the V. harveyi infection groups, the Hc content reached a maximum of 87.97 ± 4.39 mg ml(-1) at 36 h PI in the 10(6) CFU ml(-1) group, 73.74 ± 4.38 mg ml(-1) and 72.47 ± 2.09 mg ml(-1) at 12 h PI in the 10(7) and 10(8) CFU ml(-1) groups, respectively. TPCH reached a maximum of 111.16 ± 0.86 mg ml(-1) at 36 h PI in the 10(6) CFU ml(-1) group, 100.41 ± 0.51 mg ml(-1) and 101.94 ± 0.47 mg ml(-1) at 12 h PI in the 10(7) and 10(8) CFU ml(-1) groups, respectively. These data showed that both Hc content and TPCH varied as the same extent after infection. The up-regulation of the Hc content at 6-36 h PI might be a reference threshold for shrimp infection.

  3. Development of a monoclonal antibody-based flow-through immunoassay (FTA) for detection of white spot syndrome virus (WSSV) in black tiger shrimp Penaeus monodon.

    Patil, R; Shankar, K M; Kumar, B T N; Kulkarni, A; Patil, P; Moger, N

    2013-09-01

    A flow-through immunoassay (FTA), an improved version of immunodot, was developed using a nitrocellulose membrane baked onto adsorbent pads enclosed in a plastic cassette to detect white spot syndrome virus (WSSV) in shrimp. Sharp purple dots developed with WSSV against the white background of the nitrocellulose membrane. The detection limits of WSSV by the FTA and immunodot were 0.312 and 1.2 μg mL(-1) crude WSSV protein, respectively. The FTA could be completed in 8-10 min compared with 90 min for immunodot. The FTA was 100 times more sensitive than 1-step polymerase chain reaction (PCR) and in between that of the 1- and 2-step PCR protocol recommended by the Office of International Epizootics (OIE). In experimental, orally infected shrimp post-larvae, WSSV was first detected 14, 16 and 18 h post-infection (hpi) by FTA, immunodot and one-step PCR, respectively. The FTA detected WSSV 2 and 4 h earlier than immunodot and one-step PCR, respectively. The FTA was more sensitive (25/27) than one-step PCR (23/27) and immunodot (23/27) for the detection of WSSV from white spot disease outbreak ponds. The reagent components of the FTA were stable giving expected results for 6 m at 4-8 °C. The FTA is available as a rapid test kit called 'RapiDot' for the early detection of WSSV under field conditions.

  4. 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.

    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

  5. Interaction between Kazal serine proteinase inhibitor SPIPm2 and viral protein WSV477 reduces the replication of white spot syndrome virus.

    Ponprateep, Sirikwan; Phiwsaiya, Kornsunee; Tassanakajon, Anchalee; Rimphanitchayakit, Vichien

    2013-09-01

    White spot syndrome (WSS) is a viral disease caused by white spot syndrome virus (WSSV) which leads to severe mortality in cultured penaeid shrimp. In response to WSSV infection in Penaeus monodon, a Kazal serine proteinase inhibitor SPIPm2, normally stored in the granules of granular and semi-granular hemocytes is up-regulated and found to deter the viral replication. By using yeast two-hybrid screening, we have identified a viral target protein, namely WSV477. Instead of being a proteinase, the WSV477 was reported to be a Cys2/Cys2-type zinc finger regulatory protein having ATP/GTP-binding activity. In vitro pull down assay confirmed the protein-protein interaction between rSPIPm2 and rWSV477. Confocal laser scanning microscopy demonstrated that the SPIPm2 and WSV477 were co-localized in the cytoplasm of shrimp hemocytes. Using RNA interference, the silencing of WSV477 resulted in down-regulated of viral late gene VP28, the same result obtained with SPIPm2. In this instance, the SPIPm2 does not function as proteinase inhibitor but inhibit the regulatory function of WSV477.

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

    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.

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

    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.

  8. Expression profile of key immune-related genes in Penaeus monodon juveniles after oral administration of recombinant envelope protein VP28 of white spot syndrome virus.

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

    2016-07-01

    White spot syndrome virus (WSSV) is the most catastrophic pathogen the shrimp industry has ever encountered. VP28, the abundant envelope protein of WSSV was expressed in bacteria, the purified protein administered orally to Penaeus monodon juveniles and its immune modulatory effects examined. The results indicated significant up-regulation of caspase, penaeidin, crustin, astakine, syntenin, PmRACK, Rab7, STAT and C-type lectin in animals orally administered with this antigen. This revealed the immune modulations in shrimps followed by oral administration of rVP28P which resulted in the reduced transcription of viral gene vp28 and delay in mortality after WSSV challenge. The study suggests the potential of rVP28P to elicit a non-specific immune stimulation in shrimps.

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

    Yun-Long Tsai

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

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

    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 using 700 L. vannamei of known WSSV infection status shows that the established assayhassensitivity93.5% (95% CI: 90.61-95.56%) and specificity 97% (95% CI: 94.31-98.50%). Furthermore, no discrepancy was found between the two assays when 100 random L. vannamei samples were tested in parallel. Finally, excellent correlation was observed among test results of three batches of reagents with 64 samples analyzed in three different laboratories. Working in a portable device, IQ Plus™ WSSV Kit with POCKIT system allows reliable, sensitive and specific on-site detection of WSSV in L. vannamei.

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

    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

  12. HST image of Saturn's 'white spot'

    1990-01-01

    Saturn's 'white spot' or cloud believed to be ammonia ice crystals recorded by the Hubble Space Telescope (HST) planetary camera in blue and infrared light. HST data was computer-processed improving the image sharpness.

  13. White-spot disease of salmon fry

    Mazuranich, J.J.; Nielson, W.E.

    1959-01-01

     White-spot disease, sometimes referred to as coagulated-yolk disease, has been associated with excessive mortalities occurring among the fry and early fingerling stages of the fall chinook salmon (Oncorhynchus tshawytacha) at the U.S. Fish-Cultural Stations at Carson, Cook, Underwood, and Willard, Washington. This disease of eggs and fry should not be confused with the "white-spot" infection that is caused in fingerlings by members of the protozoan genus Ichthyophthirius.

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

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

    2013-01-01

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

  15. Per os infectivity of white spot syndrome virus (WSSV) in white-legged shrimp (Litopenaeus vannamei) and role of peritrophic membrane.

    Thuong, Khuong Van; Tuan, Vo Van; Li, Wenfeng; Sorgeloos, Patrick; Bossier, Peter; Nauwynck, Hans

    2016-02-29

    As earlier observations on peroral infectivity of WSSV in white-legged shrimp are conflicting, here, a standardized peroral intubation technique was used to examine (i) the role of the physical composition of the viral inoculum and (ii) the barrier function of the PM. In a first experiment, the infectivity of a WSSV stock was compared by determining the SID50 by intramuscular injection, peroral inoculation or via feeding. The following titers were obtained: 10(8.77) SID50/g by intramuscular injection, 10(1.23) SID50/g by peroral inoculation and 10(0.73) SID50/g by feeding. These results demonstrated that 10(7.54)-10(8.03) infectious virus is needed to infect shrimp by peroral inoculation and via feeding. Next, it was examined if damage of the PM may increase the susceptibility for WSSV by peroral route. The infectivity of a virus stock was tested upon peroral inoculation of shrimp with and without removal of the PM and compared with the infectivity upon intramuscular inoculation. The virus titers obtained upon intramuscular injection and peroral inoculation of shrimp with and without PM were 10(8.63), 10(1.13) and 10(1.53) SID50/mL, respectively. This experiment confirmed the need of 10(7.1)-10(7.5) infectious virus to infect shrimp via peroral route and showed that the removal of the PM slightly but not significantly (p > 0.05) facilitated the infection of shrimp. This study indicated that WSSV contaminated feed is poorly infectious via peroral route, whereas it is highly infectious when injected into shrimp. The PM plays a minor role as internal barrier of shrimp against WSSV infection.

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

    César M. Escobedo-Bonilla

    2015-04-01

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

  17. Sulfated galactans isolated from the red seaweed Gracilaria fisheri target the envelope proteins of white spot syndrome virus and protect against viral infection in shrimp haemocytes.

    Rudtanatip, Tawut; Asuvapongpatana, Somluk; Withyachumnarnkul, Boonsirm; Wongprasert, Kanokpan

    2014-05-01

    The present study was aimed at evaluating an underlying mechanism of the antiviral activity of the sulfated galactans (SG) isolated from the red seaweed Gracilaria fisheri against white spot syndrome virus (WSSV) infection in haemocytes of the black tiger shrimp Penaeus monodon. Primary culture of haemocytes from Penaeus monodon was performed and inoculated with WSSV, after which the cytopathic effect (CPE), cell viability and viral load were determined. Haemocytes treated with WSSV-SG pre-mix showed decreased CPE, viral load and cell mortality from the viral infection. Solid-phase virus-binding assays revealed that SG bound to WSSV in a dose-related manner. Far Western blotting analysis indicated that SG bound to VP 26 and VP 28 proteins of WSSV. In contrast to the native SG, desulfated SG did not reduce CPE and cell mortality, and showed low binding activity with WSSV. The current study suggests that SG from Gracilaria fisheri elicits its anti-WSSV activity by binding to viral proteins that are important for the process of viral attachment to the host cells. It is anticipated that the sulfate groups of SG are important for viral binding.

  18. PmTBC1D20, a Rab GTPase-activating protein from the black tiger shrimp, Penaeus monodon, is involved in white spot syndrome virus infection.

    Yingvilasprasert, Wanchart; Supungul, Premruethai; Tassanakajon, Anchalee

    2014-02-01

    TBC (TRE2/BUB2/CDC16) domain proteins contain an ≈ 200-amino-acid motif and function as Rab GTPase-activating proteins that are required for regulating the activity of Rab proteins, and so, in turn, endocytic membrane trafficking in cells. TBC domain family member 20 (TBC1D20) has recently been reported to mediate Hepatitis C virus replication. Herein, PmTBC1D20 identified from the black tiger shrimp, Penaeus monodon, was characterized and evaluated for its role in white spot syndrome virus (WSSV) infection. The full-length cDNA sequence of PmTBC1D20 contains 2003 bp with a predicted 1443 bp open reading frame encoding a deduced 480 amino acid protein. Its transcript levels were significantly up-regulated at 24 and 48 h by ≈ 2.3- and 2.1-fold, respectively, after systemic infection with WSSV. In addition, depletion of PmTBC1D20 transcript in shrimps by double stranded RNA interference led to a decrease in the level of transcripts of three WSSV genes (VP28, ie1 and wsv477). This suggests the importance of PmTBC1D20 in WSSV infection. This is the first report of TBC1D20 in a crustacean and reveals the possible mechanism used by WSSV to modulate the activity of the host protein, PmTBC1D20, for its benefit in viral trafficking and replication.

  19. Comparative transcriptome and potential antiviral signaling pathways analysis of the gills in the red swamp crayfish, Procambarus clarkii infected with White Spot Syndrome Virus (WSSV

    Zhi-Qiang Du

    Full Text Available Abstract Red swamp crayfish is an important model organism for research of the invertebrate innate immunity mechanism. Its excellent disease resistance against bacteria, fungi, and viruses is well-known. However, the antiviral mechanisms of crayfish remain unclear. In this study, we obtained high-quality sequence reads from normal and white spot syndrome virus (WSSV-challenged crayfish gills. For group normal (GN, 39,390,280 high-quality clean reads were randomly assembled to produce 172,591 contigs; whereas, 34,011,488 high-quality clean reads were randomly assembled to produce 182,176 contigs for group WSSV-challenged (GW. After GO annotations analysis, a total of 35,539 (90.01%, 14,931 (37.82%, 28,221 (71.48%, 25,290 (64.05%, 15,595 (39.50%, and 13,848 (35.07% unigenes had significant matches with sequences in the Nr, Nt, Swiss-Prot, KEGG, COG and GO databases, respectively. Through the comparative analysis between GN and GW, 12,868 genes were identified as differentially up-regulated DEGs, and 9,194 genes were identified as differentially down-regulated DEGs. Ultimately, these DEGs were mapped into different signaling pathways, including three important signaling pathways related to innate immunity responses. These results could provide new insights into crayfish antiviral immunity mechanism.

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

    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.

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

    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. White spot syndrome virus IE1 and WSV056 modulate the G1/S transition by binding to the host retinoblastoma protein.

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

    2013-12-01

    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.

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

    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 (α2M), 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.

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

    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. Development of primary cell cultures from mud crab, Scylla serrata, and their potential as an in vitro model for the replication of white spot syndrome virus.

    Deepika, A; Makesh, M; Rajendran, K V

    2014-01-01

    Primary cell cultures were developed from haemocytes and testis of Scylla serrata. Haemocytes were collected from live animals and cultured in double-strength L-15 medium (2× L-15) prepared in crab saline, supplemented with 5% foetal bovine serum and antibiotic-antimycotic solution (penicillin 100 U/mL, streptomycin 100 μg/mL and amphotericin B 0.25 μg/mL) with osmolality adjusted to 894 mOsm/kg. The haemocytes adhered within 2 h after seeding and showed proliferation up to 72 h. The disaggregated testis tissue fragments were seeded in 3× L-15 supplemented with non-essential amino acid mixture, lipid concentrate and antibiotic-antimycotic solution, with osmolality adjusted to 1,035 mOsm/kg with crab saline. Cells from the testis could be subcultured and maintained up to 21 d as suspension culture. Different dilutions of white spot syndrome virus (WSSV) inoculum (known virus copy number) prepared from infected Penaeus monodon were inoculated in the cultured cells, and the cytopathic effects like detachment, rounding of cells and clear areas of depleted cells were observed after 48 h in haemocyte cultures. However, WSSV-exposed testis cells did not show any obvious change until 72 h post-infection. WSSV was detected in both haemocyte and testis cultures at different time-points of infection by conventional and real-time PCR using WSSV-specific primers. The transcripts of WSSV were found to be much higher in haemocytes than in testis culture. The virus harvested from the cultured haemocytes after three passages could infect healthy P. monodon. The present study showed that mud crab haemocyte culture can support WSSV replication, and it can be used as an in vitro tool for WSSV replication.

  6. Immunostimulatory activity of sulfated galactans isolated from the red seaweed Gracilaria fisheri and development of resistance against white spot syndrome virus (WSSV) in shrimp.

    Wongprasert, Kanokpan; Rudtanatip, Tawut; Praiboon, Jantana

    2014-01-01

    Sulfated galactans (SG) were isolated from the red seaweed Gracilaria fisheri (G. fisheri). Chemical analysis revealed SG contains sulfate (12.7%) and total carbohydrate (42.2%) with an estimated molecular mass of 100 kDa. Structure analysis by NMR and FT-IR spectroscopy revealed that SG is a complex structure with a linear backbone of alternating 3-linked β-D-galactopyranose and 4-linked 3,6-anhydrogalactose units with partial 6-O-methylate-β-D-galactopyranose and with sulfation occurring on C4 of D-galactopyranose and C6 of L-galactopyranose units. SG treatment enhanced immune parameters including total haemocytes, phenoloxidase activity, superoxide anions and superoxide dismutase in shrimp Penaeus monodon. Shrimp fed with Artemia salina enriched with SG (100 and 200 μg ml(-1)) and inoculated with white spot syndrome virus (WSSV) showed a significantly lower mortality rate and lower viral VP 28 amplification and expression than control. The results suggest that SG from G. fisheri exhibits immune stimulatory and antiviral activities that could protect P. monodon from WSSV infection.

  7. Effect of ingestion and waterborne routes under different shrimp densities on white spot syndrome virus susceptibility in three commercially important penaeid shrimps

    Kuzhanthaivel Raja

    2015-11-01

    Full Text Available A study was undertaken to evaluate some factors that are important for better management of the white spot syndrome virus (WSSV. We examined the likelihood of that the mud crab (Scylla serrata is a potential carrier of WSSV in shrimp farms and then compared susceptibility of three shrimp species to WSSV, and finally tested the effects of shrimp stocking density and transmission routes on the intensity of WSSV infection under experimental conditions. Mud crabs were collected from the crab fattening unit of Pichavaram mangroves area, southeast coast of India. The infectivity test was conducted on Penaeus monodon, P. indicus, Litopenaeus vannamei under three different shrimp densities and two different routes of infection. Each treatment was executed in triplicate of 100 L tanks. The results showed that S. serrata is a carrier of WSSV. Among three species of shrimps, P. indicus was most susceptible and L. vannamei was most resistant in term of cumulative mortality. Ingestion route was more effective than waterborne route on WSSV outbreaks. Cumulative mortality of shrimps due to WSSV infection increased with increasing shrimp density. The result of this study is important to improve management especially to minimize the WSSV infection in the shrimp farms.

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

    Chen, Rong-Yuan; Shen, Kai-Li; Chen, Zhen; Fan, Wei-Wei; Xie, Xiao-Lu; Meng, Chuang; Chang, Xue-Jiao; Zheng, Li-Bing; Jeswin, Joseph; Li, Cheng-Hua; Wang, Ke-Jian; Liu, Hai-Peng

    2016-07-07

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

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

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

    2016-03-01

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

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

    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.

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

    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.

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

    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 IκB kinases IKKα and IKKβ and the IKK-related kinases TANK-binding kinase 1 (TBK1) and IKKε are the master regulators of the NF-κB 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, LvIKKβ and LvIKKε, from Pacific white shrimp, Litopenaeus vannamei. LvIKKβ and LvIKKε mRNAs are widely expressed in different tissues and are responsive to white spot syndrome virus (WSSV) infection. When overexpressed in Drosophila S2 cells, LvIKKβ but not LvIKKε activates the promoters of NF-κB pathway-controlled antimicrobial peptide genes (AMPs), such as the Penaeidins (PENs). In HEK 293T cells, both LvIKKβ and LvIKKε activate an NF-κB reporter. The silencing of LvIKKβ or LvIKKε using double-stranded RNA (dsRNA)-mediated RNA interference (RNAi) decreases the expression of L. vannamei AMPs, including PENs, lysozyme and crustins. Intriguingly, LvIKKβ- or LvIKKε-silenced L. vannamei are resistant to WSSV infection. We hypothesized that successful infection with WSSV requires the activation of the IKK-NF-κB 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 LvIKKβ or LvIKKε in Drosophila S2 cells. Taken together, our results reveal that LvIKKβ and LvIKKε may participate in the regulation of shrimp AMPs and that WSSV may subvert the L. vannamei IKK-NF-κB signaling pathway to facilitate viral gene expression.

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

    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 β subunit of F1-ATP synthase (ATPsyn-β) 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-β 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.

  14. Dietary supplementation of probiotic Bacillus PC465 isolated from the gut of Fenneropenaeus chinensis improves the health status and resistance of Litopenaeus vannamei against white spot syndrome virus.

    Chai, Peng-Cheng; Song, Xiao-Ling; Chen, Guo-Fu; Xu, Hua; Huang, Jie

    2016-07-01

    This study conducted a 30-day feeding trial and a subsequent 20-day anti-virus infection trial to determine the effects of probiotic Bacillus PC465 on the growth, health status, and disease resistance of Litopenaeus vannamei. Shrimp samples were fed with three practical diets prepared from shrimp feed containing varying probiotic doses [0 (control), 10(7), and 10(9) CFU g(-1)]. Probiotic supplementation significantly increased the weight gain and survival of L. vannamei (p vannamei. Likewise, probiotic treatment increased the transcription of hemocyanin in the hepatopancreas of L. vannamei. The probiotic treatment also significantly increased the transcription of prophenoloxidase (proPO) but decreased the transcription of crustin in hemocytes. By contrast, the same treatment failed to increase the transcription of Ras-related protein (Rab-6) in hemocytes. The number of species and biomass of Bacillus in the mid-gut were higher in the probiotic-fed group than in the control group. The total biomass of microbes was higher in the shrimp fed with 10(7) CFU g(-1) than in the shrimp fed with 10(9) CFU g(-1) and the control group on days 15 and 30 post-feeding. In two white spot syndrome virus (WSSV) infections, the weight gain, survival, and WSSV copies within the gills of the probiotic-treated shrimp significantly differed (p vannamei. Probiotic treatment also enhances the microbial structures in the gut, promotes the immune status of shrimp, and provides protection against viral infection. The supplementation with 10(9) CFU g(-1) can also improve the growth and survival of L. vannamei.

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

    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.

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

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

    2016-04-01

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

  17. Primary hemocyte culture of Penaeus monodon as an in vitro model for white spot syndrome virus titration, viral and immune related gene expression and cytotoxicity assays.

    Jose, Seena; Mohandas, A; Philip, Rosamma; Bright Singh, I S

    2010-11-01

    Immortal cell lines have not yet been reported from Penaeus monodon, which delimits the prospects of investigating the associated viral pathogens especially white spot syndrome virus (WSSV). In this context, a method of developing primary hemocyte culture from this crustacean has been standardized by employing modified double strength Leibovitz-15 (L-15) growth medium supplemented with 2% glucose, MEM vitamins (1×), tryptose phosphate broth (2.95 gl⁻¹), 20% FBS, N-phenylthiourea (0.2 mM), 0.06 μg ml⁻¹ chloramphenicol, 100 μg ml⁻¹ streptomycin and 100 IU ml⁻¹ penicillin and hemolymph drawn from shrimp grown under a bio-secured recirculating aquaculture system (RAS). In this medium the hemocytes remained viable up to 8 days. 5-Bromo-2'-deoxyuridine (BrdU) labeling assay revealed its incorporation in 22 ± 7% of cells at 24h. Susceptibility of the cells to WSSV was confirmed by immunofluorescence assay using a monoclonal antibody against 28 kDa envelope protein of WSSV. A convenient method for determining virus titer as MTT(50)/ml was standardized employing the primary hemocyte culture. Expression of viral genes and cellular immune genes were also investigated. The cell culture could be demonstrated for determining toxicity of a management chemical (benzalkonium chloride) by determining its IC(50). The primary hemocyte culture could serve as a model for WSSV titration and viral and cellular immune related gene expression and also for investigations on cytotoxicity of aquaculture drugs and chemicals.

  18. 赤潮异弯藻对中国对虾感染白斑综合症病毒的影响%The effects of Heterosigma akashiwo on artificial infection of white spot syndrome virus in Fenneropenaeus chinensis

    李才文; 管越强; 于仁诚; 张波; 俞志明

    2003-01-01

    @@ 1引言 白斑综合症病毒(white spot syndrome virus,WSSV)是近年来危害我国乃至世界对虾养殖业的主要流行病原[1~4].WSSV病毒暴发流行范围广、致病性强,自出现以来给对虾养殖业造成了巨大的损失.

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

    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.

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

    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.

  1. Advances of proteins interaction involved in white spot syndrome virus infection%白斑综合症病毒感染相关蛋白质相互作用的研究进展

    左华丽; 金春英; 王蔚

    2011-01-01

    白斑综合症病毒(white spot syndrome virus,WSSV)是危害对虾的主要病原,给全球水产养殖业带来了巨大经济损失,但至今仍未发现有效的防治方法.研究病毒与宿主的相互作用对于深入了解病毒的致病机理和宿主的免疫机制,从而寻找合适的抗病毒措施具有非常重要的理论意义和实际应用价值.该文主要介绍了蛋白质相互作用的研究方法,以及WSSV病毒蛋白之间、病毒一宿主蛋白之间和宿主蛋白之间相互作用的研究进展,为有效地防治WSSV及相关科研提供参考.%White spot syndrome virus (WSSV) is the causative agent of white spot syndrome disease of cultured penaeid shrimp. It causes high mortalities and severe damage to the global shrimp culture industry. So far, there is no efficient approach to control this virus. Research on the interaction of proteins encoded by virus and host can help to understand the pathopoiesis mechanism of virus infection and immune system of the host. The knowledge can provide promising strategies for protection and treatment of viral disease. This article mainly presented the approaches employed to analyze protein-protein interaction, the advances of protein interactions involved in WSSV infection, aiming to provide a useful reference for the disease control and the interrelated research.

  2. Jupiter Great Red Spot and White Ovals

    1979-01-01

    This photo of Jupiter was taken by Voyager 1 on March 1, 1979. The spacecraft was 3 million miles (5 million kilometers) from Jupiter at the time. The photo shows Jupiter's Great Red Spot (upper right) and the turbulent region immediately to the west. At the middle right of the frame is one of several white ovals seen on Jupiter from Earth. The structure in every feature here is far better than has ever been seen from any telescopic observations. The Red Spot and the white oval both reveal intricate and involved structure. The smallest details that can be seen in this photo are about 55 miles (95 kilometers) across. JPL manages and controls the Voyager project for NASA's Office of Space Science.

  3. Jupiter's Great Red Spot and White Ovals

    1979-01-01

    This photo of Jupiter was taken by Voyager 1 on the evening of March 1, 1979, from a distance of 2.7 million miles (4.3 million kilometers). The photo shows Jupiter's Great Red Spot (top) and one of the white ovals than can be seen in Jupiter's atmosphere from Earth. The white ovals were seen to form in 1939, and 1940, and have remained more or less constant ever since. None of the structure and detail evident in these features have ever been seen from Earth. The Great Red Spot is three times as large as Earth. Also evident in the picture is a great deal of atmospheric detail that will require further study for interpretation. The smallest details that can be seen in this picture are about 45 miles (80 kilometers across. JPL manages and controls the Voyager project for NASA's Office of Space Science.

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

    Xiaoqian Tang

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

  5. 对虾白斑杆状病毒(WSBV)定量PCR检测技术研究%Research of Quantitative PCR Detection Technology of White Spot Bacilliform Virus (WSBV)

    徐丽美; 王玮; 杨丰

    2001-01-01

    A DNA fragment of White Spot Bac illiform Virus which is lack of 74 bp was preperred and used as a competi tive internal sta ndard DNA which is amplified with the positive template or infected shrimp DNA tem plate to study its recovery and determine the content of WSBV in the infected sh rimp. The results showed that the sensitivity of the quantitave PCR reach to the Virus DNA of 10 molecular/μl. The recovery was estimated at about 50%.%报道了利用DNA重组技术在一段对虾白斑杆状病毒(White Spot Bacilliform Viru s,, WSBV)的基因组DNA片段中缺失74bp,重组得到的内标质粒作为竞争性内标模板,并与 阳标模板或病虾DNA模板共同扩增,建立了定量PCR检测方法。研究表明,定量PCR方法检测 灵敏度达10病毒分子DNA/μl,样品DNA模板制备回收率约50%,并能对病虾样品中的WSBV含 量进行定量分析。

  6. 对虾白斑综合征病毒研究进展%Research progresses of shrimp white spot syndrome virus

    魏克强; 许梓荣

    2005-01-01

    白斑综合征(white spot syndrome,WSS)是全球对虾养殖业所面临的危害性最大的病害之一,是国际兽医局(OIE)、联合国粮农组织(FAO)及亚太地区水产养殖发展网络中心(NACA)规定报告的重要水生动物疾病。自20世纪90年代初首先在我国台湾地区发生以来,至今已席卷了世界各国的对虾养殖地区,造成了巨大的经济损失,

  7. Research on Immune Protection of White Spot Syndrome Virus Surface Antigen Gene%对虾白斑综合征病毒表面抗原基因免疫保护性的研究

    张娜; 刘骁

    2015-01-01

    In this report, we constructed multivalent DNA vaccine by the combination of gene V P19、V P28 and tested their protective effects on the shrimp against the challenge of white Spot Syndrome Virus (wSSV ). Theresults indicated that DNA vaccines could protect shrimp fromwSSV infection. Thebest one of themwas PV AX 1.0-V P19-V P28 multivalent DNA vaccine.%将对虾白斑综合征病毒(White Spot Syndrome Virus,W SSV)的VP19、VP28基因融合克隆入pVAX1.0真核表达载体制备成多价 DNA 疫苗,并用构建的 DNA 疫苗制备成饲料免疫实验对虾,用RT-PCR 检测DNA 疫苗在对虾体内的表达,同时比较构建的多价DNA 疫苗与VP19、VP28单价DNA 疫苗对抗W SSV的保护率。结果显示构建的DNA疫苗有抗W SSV免疫保护性,且pVAX1.0-VP19-VP28多价DNA 疫苗保护率高于单价疫苗。

  8. Sustainable control of white spot disease

    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. C...... 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........ 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...

  9. Summary of Research on Structural Protein of White Spot Syndrome Virus in Shrimp%对虾白斑综合征病毒结构蛋白的研究概况

    李明; 张言召; 韩振乾

    2013-01-01

    白斑综合征病毒(White spot syndrome virus,WSSV)是严重危害对虾养殖业的病原之一.随着WSSV基因组全序列的公布及蛋白质组学的飞速发展,对WSSV结构蛋白及其相互作用的研究越来越深入.通过对WSSV结构蛋白及其相互作用的分析,进一步研究其在病毒感染过程中的作用,这为有效控制甚至阻断WSSV感染奠定基础.

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

    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

  11. Restricted spread of tomato spotted wilt virus

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

    2003-01-01

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

  12. Duodenal White Spots Mimicking Intestinal Candidiasis: Report of Case

    Ozgur Turk

    2015-12-01

    Full Text Available Duodenal white spots are mentioned in these nonspecific lesions until recently. Although there is not enough studies about duedonal white spots yet; these lesions described in a separate syndrome. Here now we reported a case that we diagnosed multiple Duodenal white spots mimicking intestinal candidiasis. Clinical manifestation and endoscopic appearance of lesions gave rise to thought intestinal candidiasis histopathological examination revealed us several duodenitis. There was no evidence of fungal infection in PAS staining. Early after endoscopy patient took treatment of Lansoprozole at the 30 mg dose and bismuth salicylate. Patients compliant declined and control endoscopy exposed white duodenal spots cleared away three months later. Duodenal white spots are becoming to be designated as a separate disease even a syndrome. Etiology of duodenal white spots must be determined carefully during endoscopy. Sometimes it is difficult to make the correct diagnosis by appearance of lesion; in such cases histopathological examination can be useful both differential diagnosis of disease and determination of etiological factor. [J Contemp Med 2015; 5(4.000: 249-252

  13. Advancements in Molecular Biology on White Spot Syndrome Virus%对虾白斑综合症病毒的分子生物学研究进展

    程伟; 刘志昕

    2005-01-01

    对虾白斑综合症病毒(wkite spot syndrome virus,WSSV)又称对虾白斑杆状病毒(white spot bacilliform virus WSBV),是对全球对虾养殖业危害最大的病原之一。该病毒分子量较大、结构特异,是已知动物病毒中最大的病毒。自1992年以来,各国学者对WSSV进行研究。由于他们研究的方法及研究的侧重点不同,对该病毒命名各异。陈秀男(1993)称之为白斑综合症杆状病毒(WSBV);荷兰Wongteerasupaya(1995)称其为系统外胚层和中胚层杆状病毒(SEMBV);

  14. Photometry of two DQ white dwarfs - search for spots

    Vornanen, Tommi; 10.1051/0004-6361/201219255

    2012-01-01

    Aims. The intensity profiles of the C_2 Swan bands in cool DQ white dwarfs cannot be adequately fitted with models that otherwise succesfully reproduce spectral features of the molecule CH in these stars. Initial modelling showed that a two-component atmosphere in the style of a spot might be able to solve the problem. Methods. We photometrically observed the two cool DQ white dwarfs GJ1117 and EGGR78 to search for variability caused by stellar spots. Results. We have not found any such variability, but we estimate the effects of hypothetical spots on lightcurves. We also estimate detection probabilities for spots in different configurations. Alternative explanations of the problem are needed and briefly discussed.

  15. [White spot lesions and orthodontic treatment. Prevention and treatment].

    Morrier, Jean-Jacques

    2014-09-01

    Decalcification of the enamel surface adjacent to fixed orthodontic appliances, in the form of white spot lesions, is a common and frequent well-known side-effect of orthodontic treatment. Fixed appliances and the bonding materials increase the retention of biofilm and encourage the formation of white spot lesions. Management of these lesions begins with a good oral hygiene regime and needs to be associated with use of fluoride agents (fluoridated toothpaste, fluoride containing mouth rinse, gel, varnish, bonding materials, elastic ligature), CPP-ACP, antiseptics, LASER, tooth whitening, resin infiltration, micro-abrasion. The purpose of this review is to access the direct evidence regarding the prevention and management of white spot lesions during and after orthodontic treatment.

  16. Distribution of White Spots after Debanding in Orthodontic Patients

    F. Arbabzadeh Zavareh

    2006-12-01

    Full Text Available Statement of problem: Fixed orthodontic appliances can interfere with removing bacterial plaques from dental surfaces which can ultimately lead to white spot formation.Purpose: The aim of this study was to evaluate the quantity of white spots and areas of decalcification following fixed orthodontic treatment.Materials and Methods: A total of 100 patients undergoing or scheduled for fixed orthodontic treatment were divided into two groups. Group A consisted of fifty volunteers before the initiation of therapy and group B included fifty patients at the end of their treatment. In group A, the buccal surfaces of the first molars in each quadrant were examined for the presence of enamel decalcifications. After removing the orthodontic bands in group B, the buccal surfaces of the first molars in each quadrant were examined for white spots. Gender, oral hygiene level, plaque index, type of cement and duration of treatment were recorded for all patients. Binomial logistic regression, chi-square and Mann-Whitney U-tests were used for statistical analysis.Results: The number of white spots in group A was 28 (14% which was significantly lower than group B with 83 (41.5% decalcified lesions (P<0.01. Oral hygiene and duration of treatment had a significant effect on the occurrence of white spots (P<0.05.Conclusion: The results showed that patients with orthodontic bands are at higher risk for white spot formation and good oral hygiene demonstrates a protective effect on their reduction. Both professional and daily oral hygiene measures can decrease thecariogenicity of bacterial plaques in these patients.

  17. 凡纳对虾白斑综合征病毒的检测和预防%Detection and prevention of the White Spot Syndrome Virus in Penaeus vannamei

    蔡生力; 陈专静

    2001-01-01

    @@ 20世纪90年代以来,日本、泰国、中国等许多亚洲国家和地区因虾病流行,对虾养殖产量锐减,造成重大经济损失.其中危害最严重的就是白斑综合征(white spot syndrome,WSS),其病原体为白斑综合征病毒(white spot syndrome virus,WSSV)[1-3].1993年5-8月,我国沿海从南到北对虾养殖场发生了大面积的对虾爆发性流行病,损失非常严重[4,5].关于WSS的研究已成为近年来国内外水产养殖病害研究的热点[6-12].随着WSSV研究的深入,有关WSS的病原、病理、诊断方法、检测技术、传播途径及综合防治技术等方面取得了较大的进展,对养殖生产的恢复起到了一定的作用.凡纳对虾( Penaeusvannamei )又名南美白对虾,是迄今所知世界养殖产量最高的三种对虾之一,我国1988年首次从美洲引进凡纳对虾,育苗和养殖在我国南北方逐步展开[13].近两年,随着育苗技术的不断成熟,养殖面积和产量也不断增加,给养殖户带来可观的经济效益.

  18. Study on experimental infection of Exopalaemon carinicauda Holehuis with white spot syndrome virus%白斑综合征病毒对脊尾白虾的致病性研究

    2013-01-01

      2010年、2011年江苏沿海多数地方海水养殖中发生了脊尾白虾大规模死亡的现象,本课题通过对患暴发性流行病的脊尾白虾(Exopalaemon carinicauda)进行病毒分子生物学检测,发现病虾体内白斑综合征病毒(White Spot Syndrome Virus,简称WSSV)检测普遍呈阳性;应用发病虾组织制备的病毒粗提液,进行了人工感染实验,实验表明该病毒对脊尾白虾具有较强的致病性,可引起68.5%的死亡率。电镜结果表明,发病虾与感染病虾鳃及肝胰脏等组织器官都发生了相同的细胞病理变化,主要表现为细胞核内染色质边聚、线粒体肿胀、内嵴消失等一系列细胞病理变化,在其细胞核与细胞质中发现了具有单层囊膜结构的对虾白斑综合征病毒粒子,病毒粒子大小约为(130~170)nm×(305~405)nm,核衣壳呈子弹形,一端较细另一端较粗,完整病毒粒子由核心、衣壳和囊膜构成,分子生物学实验表明该病毒为WSSV。实验结果符合柯赫氏法则,可以初步推断WSSV感染脊尾白虾并与引发的大规模死亡有着直接关系。%White Spot Syndrome Virus (WSSV) is one of the most devastating and virulent viral agents threatening the penaeid shrimp culture industry and has been responsible for serious economic losses in shrimp farms world-wide. By the Polymerase Chain Reaction test, the Exopalaemon carinicauda Holehuis were infected by WSSV. Quantitative challenge test was used in this study to further evaluate the pathogenic potential of WSSV, the cumula-tive mortalities of WSSV-infected group reached to 68.5%in 14 days post-inoculation. Cytopathological features of the moribund shirmp and reinfected shirmp with WSSV were indentical by transmission. Cytopathological features showed that organelles were enlarged and fragmented, and numerous viruses with different size were found in nu-clear and cytoplasm. In addition, the WSSV was detected in different tissues of reinfected shrimp

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

    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 ...

  20. A DARK SPOT ON A MASSIVE WHITE DWARF

    Kilic, Mukremin; Gianninas, Alexandros; Curd, Brandon; Wisniewski, John P. [Homer L. Dodge Department of Physics and Astronomy, University of Oklahoma, 440 W. Brooks St., Norman, OK 73019 (United States); Bell, Keaton J.; Winget, D. E.; Winget, K. I. [Department of Astronomy, University of Texas at Austin, Austin, TX 78712 (United States); Brown, Warren R. [Smithsonian Astrophysical Observatory, 60 Garden St., Cambridge, MA 02138 (United States); Hermes, J. J. [Department of Physics, University of Warwick, Coventry CV4 7AL (United Kingdom); Dufour, Patrick [Institut de recherche sur les exoplanétes (iREx), Département de Physique, Université de Montréal, C.P. 6128, Succ. Centre-Ville, Montréal, QC H3C 3J7 (Canada)

    2015-12-01

    We present the serendipitous discovery of eclipse-like events around the massive white dwarf SDSS J152934.98+292801.9 (hereafter J1529+2928). We selected J1529+2928 for time-series photometry based on its spectroscopic temperature and surface gravity, which place it near the ZZ Ceti instability strip. Instead of pulsations, we detect photometric dips from this white dwarf every 38 minutes. Follow-up optical spectroscopy observations with Gemini reveal no significant radial velocity variations, ruling out stellar and brown dwarf companions. A disintegrating planet around this white dwarf cannot explain the observed light curves in different filters. Given the short period, the source of the photometric dips must be a dark spot that comes into view every 38 minutes due to the rotation of the white dwarf. Our optical spectroscopy does not show any evidence of Zeeman splitting of the Balmer lines, limiting the magnetic field strength to B < 70 kG. Since up to 15% of white dwarfs display kG magnetic fields, such eclipse-like events should be common around white dwarfs. We discuss the potential implications of this discovery on transient surveys targeting white dwarfs, like the K2 mission and the Large Synoptic Survey Telescope.

  1. A Dark Spot on a Massive White Dwarf

    Kilic, Mukremin; Bell, Keaton J; Curd, Brandon; Brown, Warren R; Hermes, J J; Dufour, Patrick; Wisniewski, John P; Winget, D E; Winget, K I

    2015-01-01

    We present the serendipitous discovery of eclipse-like events around the massive white dwarf SDSS J152934.98+292801.9 (hereafter J1529+2928). We selected J1529+2928 for time-series photometry based on its spectroscopic temperature and surface gravity, which place it near the ZZ Ceti instability strip. Instead of pulsations, we detect photometric dips from this white dwarf every 38 minutes. Follow-up optical spectroscopy observations with Gemini reveal no significant radial velocity variations, ruling out stellar and brown dwarf companions. A disintegrating planet around this white dwarf cannot explain the observed light curves in different filters. Given the short period, the source of the photometric dips must be a dark spot that comes into view every 38 min due to the rotation of the white dwarf. Our optical spectroscopy does not show any evidence of Zeeman splitting of the Balmer lines, limiting the magnetic field strength to B<70 kG. Since up to 15% of white dwarfs display kG magnetic fields, such ecli...

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

    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 orthodontic patients. Suggestions are offered in the literature for ways to prevent this condition from manifesting itself. PMID:24987641

  3. Management of post-orthodontic white spot lesions

    Sonesson, Mikael; Bergstrand, Fredrik; Gizani, Sotiria;

    2016-01-01

    BACKGROUND/OBJECTIVES: The management of post-orthodontic white spot lesions is based on remineralization strategies or a minimal-invasive camouflage of the lesions. AIM: The aim of this systematic review was to identify and assess the quality of evidence for the various clinical technologies....... Furthermore, a follow-up period of at least 8 weeks was required. The publication bias could not be assessed due to the paucity of included trials. CONCLUSIONS/CLINICAL IMPLICATIONS: There is a lack of reliable scientific evidence to support re-mineralizing or camouflaging strategies to manage post-orthodontic...

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

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

    2013-04-01

    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 orthodontic patients. Suggestions are offered in the literature for ways to prevent this condition from manifesting itself.

  5. Endogenous retrovirus insertion in the KIT oncogene determines white and white spotting in domestic cats.

    David, Victor A; Menotti-Raymond, Marilyn; Wallace, Andrea Coots; Roelke, Melody; Kehler, James; Leighty, Robert; Eizirik, Eduardo; Hannah, Steven S; Nelson, George; Schäffer, Alejandro A; Connelly, Catherine J; O'Brien, Stephen J; Ryugo, David K

    2014-08-01

    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 (FERV1) as responsible for two distinct phenotypes of the W locus, Dominant White, and white spotting. A full-length (7125 bp) FERV1 element is associated with white spotting, whereas a FERV1 long terminal repeat (LTR) is associated with all Dominant White individuals. For purposes of statistical analysis, the alternatives of wild-type sequence, FERV1 element, and LTR-only define a triallelic marker. Taking into account pedigree relationships, deafness is genetically linked and associated with this marker; estimated P values for association are in the range of 0.007 to 0.10. The retrotransposition interrupts a DNAase I hypersensitive site in KIT intron 1 that is highly conserved across mammals and was previously demonstrated to regulate temporal and tissue-specific expression of KIT in murine hematopoietic and melanocytic cells. A large-population genetic survey of cats (n = 270), representing 30 cat breeds, supports our findings and demonstrates statistical significance of the FERV1 LTR and full-length element with Dominant White/blue iris (P < 0.0001) and white spotting (P < 0.0001), respectively.

  6. A Dark Spot on a Massive White Dwarf

    Kilic, Mukremin; Gianninas, Alexandros; Bell, Keaton J.; Curd, Brandon; Brown, Warren R.; Hermes, J. J.; Dufour, Patrick; Wisniewski, John P.; Winget, D. E.; Winget, K. I.

    2015-12-01

    We present the serendipitous discovery of eclipse-like events around the massive white dwarf SDSS J152934.98+292801.9 (hereafter J1529+2928). We selected J1529+2928 for time-series photometry based on its spectroscopic temperature and surface gravity, which place it near the ZZ Ceti instability strip. Instead of pulsations, we detect photometric dips from this white dwarf every 38 minutes. Follow-up optical spectroscopy observations with Gemini reveal no significant radial velocity variations, ruling out stellar and brown dwarf companions. A disintegrating planet around this white dwarf cannot explain the observed light curves in different filters. Given the short period, the source of the photometric dips must be a dark spot that comes into view every 38 minutes due to the rotation of the white dwarf. Our optical spectroscopy does not show any evidence of Zeeman splitting of the Balmer lines, limiting the magnetic field strength to B Tecnologia e Inovação (Brazil) and Ministerio de Ciencia, Tecnología e Innovación Productiva (Argentina).

  7. Minimally invasive treatment for esthetic enhancement of white spot lesion in adjacent tooth.

    Lee, Ji-Hye; Kim, Dae-Gon; Park, Chan-Jin; Cho, Lee-Ra

    2013-08-01

    This article describes the treatment provided to a patient with the maxillary anterior teeth exhibiting severe secondary caries beneath the previous restoration and a white spot lesion on the adjacent incisor. Two implants were placed after extraction of hopeless teeth with the guided bone regeneration technique. A white spot lesion of the adjacent incisor was treated with minimally invasive treatment. This clinical report describes the multidisciplinary treatment for the white spot lesion and esthetic restoration of missing anterior teeth.

  8. Distribution of white spots after de-bonding in orthodontic patients

    Zabokova-Bilbilova, Efka; Muratovska, Ilijana; Stojanovska, Vera; Stefafanovska, E.; Dimova, Cena

    2014-01-01

    Fixed orthodontic appliances can interfere with removing bacterial plaques from dental surfaces which can ultimately lead to white spot formation. The aim of this study was to evaluate the quantity of white spots lesions in orthodontic patients treated with fixed orthodontic appliances during different treatment periods and to reveal the teeth most commonly affected by white spot lesions. Materials and Methods: A total of 30 patients undergoing or ...

  9. Analysis of tomato spotted wilt virus genome transcription

    Knippenberg, van I.C.

    2005-01-01

    Tomato spotted wilt virus (TSWV) is the type species of the genus Tospovirus within the Bunyaviridae, a family of segmented negative strand RNA viruses. Although much ground has been covered in the past two decades, many questions concerning the mechanism of replication and transcription of this imp

  10. 白斑综合征%White spot disease

    陈爱平; 江育林; 钱冬; 陈昌福; 李安兴; 黄捷; 杨冰

    2010-01-01

    @@ 白斑综合征(white spot disease,WSD)俗称白斑病,是由白斑综合征病毒引起的对虾严重传染性疫病,20世纪90年代以来-直严重威胁着全世界对虾的养殖安全.该病的特点是发病急、死亡率高,死亡速度快.病虾停止摄食,部分濒死虾甲壳出现白色斑点,白斑综合征因此而得名,但亦可不出现白斑而虾体变红.世界动物卫生组织(OIE)将其列为强制通报疫病.

  11. White spot lesions: prevention and management during the orthodontic treatment.

    Zabokova-Bilbilova, Efka; Popovska, Lidija; Kapusevska, Biljana; Stefanovska, Emilija

    2014-01-01

    The formation of white spot lesions, or enamel demineralization, around fixed orthodontic attachments is a common complication during and following fixed orthodontic treatment, which marks the result of a successfully completed case. This article is a contemporary review of the risk factors and preventive methods of these orthodontics scars. Preventive programmes must be emphasized to all orthodontic patients. The responsibility of an orthodontist is to minimize the risk of the patient having decalcification as a consequence of orthodontic treatment by educating and motivating the patients for excellent oral hygiene practice. Prophylaxis with topical fluoride application should be implemented: high-fluoride toothpastes, fluoride mouthwashes, gels and varnishes during and after the orthodontic treatment, especially for patients at high risk of caries.

  12. Late infiltration of post-orthodontic white spot lesions.

    Neuhaus, Klaus W; Graf, Martina; Lussi, Adrian; Katsaros, Christos

    2010-11-01

    White spot lesion (WSL) infiltration has been recommended immediately after debonding of orthodontic brackets. It is however not clear if established inactive WSLs can also be masked through infiltration. Orthodontic treatment of a 19-year-old patient had to be terminated prematurely due to development of multiple WSLs of varying severity. Three months after debonding, the patient presented for lesion infiltration. After etching with 15% HCl gel and re-wetting of the dried surfaces it seemed that a good outcome could be expected. Lesion infiltration led to complete masking of less severe WSLs. The visual appearance of moderate and severe WSLs was improved but they were still visible after treatment. Inactive WSLs may not represent an increased caries risk, but patients are often bothered esthetically. Infiltration by repeated etching might be a viable approach even for inactive WSLs. Controlled clinical trials are needed to investigate the long-term performance of this technique.

  13. Rapid diagnosis of vibriosis and white spot syndrome (WSS) in the culture of shrimp, Penaeus monodon in Philippines.

    Caipang, Christopher Marlowe A; Aguana, Mary Paz N

    2010-10-01

    Viral and bacterial pathogens have raised serious concerns in the sustainability of the shrimp culture industry in the Philippines. Heavy mortality associated with luminous vibriosis and white spot syndrome virus (WSSV) infection has been the major problem besetting the industry. Using published PCR protocols for the diagnosis of vibriosis and white spot syndrome virus (WSSV) disease in shrimp, we optimized these assays that could be suited to the shrimp aquaculture setting in the Philippines. Genomic DNAs of Vibrio spp. that exhibited luminescence as well as those that grew on thiosulfate citrate bile salt sucrose agar (TCBS) were used for the PCR amplification of the ribonuclease P (RNase P) gene. There was differential amplification of the RNase P gene based on the phenotypic characters of the Vibrio spp. Similar results were also obtained using direct colony PCR of the bacterial colonies. White spot syndrome virus was also detected in the infected shrimp and there were differences in the detection frequency in relation to the tissues used for PCR amplification. Duplex PCR was also optimized that could be used for simultaneous detection of these pathogens in shrimp.

  14. Non-destructive management of white spot lesions by using tooth jewelry.

    Kim, Hee-Jin; Karanxha, Lorena; Park, Su-Jung

    2012-11-01

    Although several methods including composite resin restoration and microabrasion have been used for management of white spot lesion, tooth jewelry can be considered as another noninvasive option. This case report describes the management of white spot lesions by using tooth jewelry. This report also highlights the patients' preference for tooth jewelry as an esthetic concern.

  15. A Frameshift Mutation in KIT is Associated with White Spotting in the Arabian Camel

    Holl, Heather; Isaza, Ramiro; Mohamoud, Yasmin; Ahmed, Ayeda; Almathen, Faisal; Youcef, Cherifi; Gaouar, Semir; Antczak, Douglas F.; Brooks, Samantha

    2017-01-01

    While the typical Arabian camel is characterized by a single colored coat, there are rare populations with white spotting patterns. White spotting coat patterns are found in virtually all domesticated species, but are rare in wild species. Theories suggest that white spotting is linked to the domestication process, and is occasionally associated with health disorders. Though mutations have been found in a diverse array of species, fewer than 30 genes have been associated with spotting patterns, thus providing a key set of candidate genes for the Arabian camel. We obtained 26 spotted camels and 24 solid controls for candidate gene analysis. One spotted and eight solid camels were whole genome sequenced as part of a separate project. The spotted camel was heterozygous for a frameshift deletion in KIT (c.1842delG, named KITW1 for White spotting 1), whereas all other camels were wild-type (KIT+/KIT+). No additional mutations unique to the spotted camel were detected in the EDNRB, EDN3, SOX10, KITLG, PDGFRA, MITF, and PAX3 candidate white spotting genes. Sanger sequencing of the study population identified an additional five KITW1/KIT+ spotted camels. The frameshift results in a premature stop codon five amino acids downstream, thus terminating KIT at the tyrosine kinase domain. An additional 13 spotted camels tested KIT+/KIT+, but due to phenotypic differences when compared to the KITW1/KIT+ camels, they likely represent an independent mutation. Our study suggests that there are at least two causes of white spotting in the Arabian camel, the newly described KITW1 allele and an uncharacterized mutation. PMID:28282952

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

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

    1996-01-01

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

  17. Sensitivity comparison of three PCR-based methods for white spot syndrome virus (WSSV) detection%白斑综合征病毒(WSSV)3种PCR检测方法的灵敏度比较

    肖广侠; 李战军; 孟宪红; 张庆文; 罗坤; 孔杰

    2011-01-01

    为了探讨不同PCR检测方法的灵敏度,分别利用TaqMan实时定量PCR、世界动物卫生组织(OIE)公布的巢式PCR引物(简称OIE)、黄海水产研究所种质资源与工程育种研究室(GB)设计的引物(简称GB)及2种巢式PCR对应的一步法PCR,对具有不同白斑综合征病毒(White Spot Syndrome Virus,WSSV)含量的中国明对虾(Fenneropenaeus chinensis)样品进行检测.结果显示,当使用已知病毒含量的标准品进行检测时,TaqMan实时定量PCR方法可以检测到l0个WSSV拷贝;OIE巢式PCR与GB巢式PCR方法分别可检测到104和103个WSSV拷贝;单独使用OIE巢式PCR的外引物和内引物扩增时,分别可检测到5×104和2.5×104个WSSV拷贝;单独使用GB巢式PCR的外引物和内引物进行一步法PCR扩增时,分别可检测到104和5×103个WSSV拷贝.使用上述PCR方法分别对44份未知WSSV含量的样品进行验证,定量PCR方法检测阳性率为84.09%,OIE巢式PCR与GB巢式PCR方法检测的阳性率分别为18.18%和27.27%;单独使用OIE巢式PCR的外引物和内引物扩增检测的阳性率均为15.91%;单独使用GB巢式PCR的外引物和内引物扩增检测的阳性率分别为18.18%和20.45%.根据以上结果,PCR方法检测WSSV的灵敏度由高到低依次为:定量PCR、巢式PCR、一步法PCR.%In order to explore the limitations of different PCR methods, we compared the sensitivity of three PCR methods, including TaqMan real-time PCR, Office International Des Epizooties-designed (OIE) and Genetic resource and Breeding Lab of Yellow Sea Fisheries Research Institute-designed nested PCR (GB) and one-step PCR methods, with standard Fenneropenaeus chinensis samples with serial known WSSV load.As a result, TaqMan real-time PCR could detect with certainty 10 WSSV copies per reaction, while 104and 103 copies were needed for OIE and GB nested PCR; 5× 104 and 2.5×104 copies were needed for the outer primer and inner primer amplification of OIE nested PCR; 104 and 5×103

  18. White-spot lesions and gingivitis microbiotas in orthodontic patients.

    Tanner, A C R; Sonis, A L; Lif Holgerson, P; Starr, J R; Nunez, Y; Kressirer, C A; Paster, B J; Johansson, I

    2012-09-01

    White-spot lesions (WSL) associated with orthodontic appliances are a cosmetic problem and increase risk for cavities. We characterized the microbiota of WSL, accounting for confounding due to gingivitis. Participants were 60 children with fixed appliances, aged between 10 and 19 yrs, half with WSL. Plaque samples were assayed by a 16S rRNA-based microarray (HOMIM) and by PCR. Mean gingival index was positively associated with WSL (p = 0.018). Taxa associated with WSL by microarray included Granulicatella elegans (p = 0.01), Veillonellaceae sp. HOT 155 (p gingivitis by microarray included: Gemella sanguinis (p = 0.002), Actinomyces sp. HOT 448 (p = 0.003), Prevotella cluster IV (p = 0.021), and Streptococcus sp. HOT 071/070 (p = 0.023); and levels of S. mutans (p = 0.02) and Bifidobacteriaceae (p = 0.012) by qPCR. Species' associations with WSL were minimally changed with adjustment for gingivitis level. Partial least-squares discriminant analysis yielded good discrimination between children with and those without WSL. Granulicatella, Veillonellaceae and Bifidobacteriaceae, in addition to S. mutans and S. wiggsiae, were associated with the presence of WSL in adolescents undergoing orthodontic treatment. Many taxa showed a stronger association with gingivitis than with WSL.

  19. White spot disease risk factors associated with shrimp farming practices and geographical location in Chanthaburi province, Thailand.

    Piamsomboon, Patharapol; Inchaisri, Chaidate; Wongtavatchai, Janenuj

    2015-12-09

    Over the past 2 decades, shrimp aquaculture in Thailand has been impacted by white spot disease (WSD) caused by white spot syndrome virus (WSSV). Described here are results of a survey of 157 intensive shrimp farms in Chanthaburi province, Thailand, to identify potential farm management and location risk factors associated with the occurrence of WSD outbreaks. Logistic regression analysis of the survey responses identified WSD risks to be associated with farms sharing inlet water and culturing shrimp year round and with a single owner operating more than 1 farm. The analysis also showed WSD risks to be reduced at farms that used probiotics and applied lime to pond bottoms when fallow to neutralize acidity and kill microorganisms. Regression modeling identified no association of geographical location with WSD. The data should assist shrimp farms in mitigating the effects of WSD in Thailand.

  20. 防治对虾白斑综合征病毒(WSSV)的主要措施%A Review: Prevention and Cure of Shrimp White Spot Syndrome Virus

    闫冬春

    2006-01-01

    对虾白斑综合征病毒(white spot syndrome virus,WSSV)是迄今为止危害最为严重的一种对虾病毒,发病对虾3~10d死亡率可达到100%。该病毒自1992年爆发以来,已在世界范围内传播开来,每年给对虾养殖业造成巨大经济损失,至今仍未得到完全控制,成为目前对虾养殖业可持续发展的主要障碍之一。1995年,国际兽疫局(OIE)、联合国粮农组织(FAO)以及亚太地区水产养殖发展网络中心(NACA)将其列为需要报告的水生动物病毒性疫病之一。

  1. A vector that expresses VP28 of WSSV can protect red swamp crayfish from white spot disease.

    Mu, Yi; Lan, Jiang-Feng; Zhang, Xiao-Wen; Wang, Xian-Wei; Zhao, Xiao-Fan; Wang, Jin-Xing

    2012-02-01

    White spot disease caused by white spot syndrome virus (WSSV) leads to devastating losses in shrimp farming. The WSSV envelope protein VP28, can be used as subunit vaccines that can efficiently protect shrimp against WSSV disease. However, the function of the envelope protein VP19 was not confirmed, some researches found that VP19 could protect shrimp against WSSV, and other reports found it no any protection. To detect the functions of VP28 and VP19 and find a method to prevent this disease in red swamp crayfish Procambarus clarkii, we constructed the plasmid vectors pIevp28 and pIevp19, which contains the ie1 promoter and coding region of vp28 or vp19 of WSSV, respectively. The results of quantitative real-time PCR and western blot showed that the injected vectors could transcribe corresponding mRNAs and translate to the protein VP28 or VP19 in the crayfish. The vp28 or vp19 signal was detected on the third day post injection, and maintained its expression for 30days. The mortality of the crayfish with pIevp28 showed obvious decline compared with the controls (pIe and PBS injection). However, pIevp19 seems did not affect the mortality of the crayfish compared with the controls. Furthermore, only VP28 was found tightly bound to the host haemocytes under immunocytochemistry. The results suggest that the VP28 protein might protect shrimp from the virus through competitive inhibition. We also found that oral administration of Escherichia coli with pIevp28 could protect crayfish from white spot disease, but the E. coli with pIevp19 was not. Therefore, we think that oral administration of bacteria with pIevp28 is a potentially easy therapeutic way against white spot disease in aquaculture.

  2. Pigmented spots in the wool-bearing skin on white merino sheep induced by ultraviolet light.

    Forrest, J W; Fleet, M R

    1986-01-01

    Black-grey pigmented skin spots, some of which contained pigmented wool fibres, were observed in a flock of 8.5-year-old white Merino ewes. The spots were concentrated along the backline and increased in number following shearing, suggesting exposure to sunlight to be of importance in the development of these non-congenital pigmented skin spots in genetically white Merino sheep. To test the effect of ultraviolet light, white Merino sheep, ranging in age from 3 to 8 years, had a closely clipped midside area of wool-bearing skin irradiated on each of 28 consecutive days. Pigmented skin spots developed in 6 of the 16 white Merino sheep irradiated. Spots first appeared after 10 days of irradiation, the number subsequently increasing with time, and two skin spots were found to contain sparse numbers of black-grey pigmented wool fibres. Histological examination showed both the naturally occurring and irradiation-induced pigmented skin spots resulted from an increase in both number and activity of melanocytes localized along the epidermal-dermal border of the epidermis. With time, the melanocytes were observed to have entered, to varying depths, the outer-root sheath of follicles still producing white wool fibres. These ultraviolet-light-induced changes to epidermal melanocytes in white Merino sheep presumably occur due to alterations within the local tissue environment in which the melanocytes lie.

  3. Prevention of white spot lesions in orthodontic practice: a contemporary review.

    Sudjalim, T R; Woods, M G; Manton, D J

    2006-12-01

    The development of white spot demineralization associated with fixed appliance orthodontic treatment is a significant clinical problem. Both established and experimental methods for prevention of such lesions in day-to-day clinical practice are presented and discussed.

  4. Fluoride-Releasing Materials to Prevent White Spot Lesions around Orthodontic Brackets: A Systematic Review.

    Nascimento, Patrícia Layane de Menezes Macêdo; Fernandes, Micaelle Tenório Guedes; Figueiredo, Fabricio Eneas Diniz de; Faria-E-Silva, André Luis

    2016-01-01

    The relation between orthodontic fixed appliances use and enamel demineralization is well established. Different preventive approaches have been suggested to this problem, but controversy remains about which is the best. The aim of this study was to perform a systematic review of clinical trials that investigated the effectiveness of materials containing fluorides to lute brackets or cover the bonding interface in order to inhibit the development and progression of white spot lesions. The null hypothesis was that fluoride materials do not affect the incidence of white spot lesions around brackets. A MEDLINE search was conducted for randomized clinical trials evaluating the development of white spot lesions in patients using fixed orthodontic appliances, followed by meta-analysis comparing the results for patients for whom dental materials containing fluorides were used (experimental group) to those for whom these materials were not used (control group). The pooled relative risk of developing white spot lesions for the experimental group was 0.42 (95% confidence interval: 0.25 to 0.72); hence, when fluoride-releasing materials are used, the patient has 58% less risk of white spot lesion development. Regarding white spot lesion extent, the pooled mean difference between the experimental and control groups was not statistically significant (-0.12; 95% confidence interval: -0.29 to 0.04). In conclusion, the results of the present systematic review suggest that fluoride-releasing materials can reduce the risk of white spot lesions around brackets. However, when white spot lesions had already occurred, there is no evidence that fluoride-releasing materials reduce the extent of these lesions.

  5. [Inheritance of white spots on the shell of Littorina obtusata (Gastropoda, Prosobranchia)].

    Kozminskiĭ, E V; Lezin, P A; Fokin, M V

    2010-12-01

    The hypothesis on monogenic inheritance of white spot pattern on the shell of the gastropod snail Littorina obtusata was tested. Although in most cases our results did not contradict this assumption, the hypothesis cannot explain the observed segregation in general. An alternative hypothesis was suggested, according to which the presence of spot pattern is controlled by two complementary biallelic genes.

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

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

  7. The 2010 Saturn's Great White Spot: Observations and models

    Sanchez-Lavega, A.

    2011-12-01

    On December 5, 2010, a major storm erupted in Saturn's northern hemisphere at a planetographic latitude of 37.7 deg [1]. These phenomena are known as "Great White Spots" (GWS) and they have been observed once per Saturn year since the first case confidently reported in 1876. The last event occurred at Saturn's Equator in 1990 [2]. A GWS differs from similar smaller-scale storms in that it generates a planetary-scale disturbance that spreads zonally spanning the whole latitude band. We report on the evolution and motions of the 2010 GWS and its associated disturbance during the months following the outbreak, based mainly on high quality images obtained in the visual range submitted to the International Outer Planet Watch PVOL database [3], with the 1m telescope at Pic-du-Midi Observatory and the 2.2 m telescope at Calar Alto Observatory. The GWS "head source" extinguished by June 2011 implying that it survived about 6 months. Since this source is assumed to be produced by water moist convection, a reservoir of water vapor must exist at a depth of 10 bar and at the same time a disturbance producing the necessary convergence to trigger the ascending motions. The high temporal sampling and coverage allowed us to study the dynamics of the GWS in detail and the multi-wavelength observations provide information on its cloud top structure. We present non-linear simulations using the EPIC code of the evolution of the potential vorticity generated by a continuous Gaussian heat source extending from 10 bar to about 1 bar, that compare extraordinary well to the observed cloud field evolution. Acknowledgements: This work has been funded by Spanish MICIIN AYA2009-10701 with FEDER support and Grupos Gobierno Vasco IT-464-07. The presentation is done on behalf of the team listed in Reference [1]. [1]Sánchez-Lavega A., et al., Nature, 475, 71-74 (2011) [2]Sánchez-Lavega A., et al., Nature, 353, 397-401 (1991) [3]Hueso R., et al., Planet. Space Sci., 58, 1152-1159 (2010).

  8. Susceptibility to infection and pathogenicity of White Spot Disease (WSD) in non-model crustacean host taxa from temperate regions.

    Bateman, K S; Tew, I; French, C; Hicks, R J; Martin, P; Munro, J; Stentiford, G D

    2012-07-01

    Despite almost two decades since its discovery, White Spot Disease (WSD) caused by White Spot Syndrome Virus (WSSV) is still considered the most significant known pathogen impacting the sustainability and growth of the global penaeid shrimp farming industry. Although most commonly associated with penaeid shrimp farmed in tropical regions, the virus is also able to infect, cause disease and kill a wide range of other decapod crustacean hosts from temperate regions, including lobsters, crabs, crayfish and shrimp. For this reason, WSSV has recently been listed in European Community Council Directive 2006/88. Using principles laid down by the European Food Safety Authority (EFSA) we applied an array of diagnostic approaches to provide a definitive statement on the susceptibility to White Spot Syndrome Virus (WSSV) infection in seven ecologically or economically important crustacean species from Europe. We chose four marine species: Cancer pagurus, Homarus gammarus, Nephrops norvegicus and Carcinus maenas; one estuarine species, Eriocheir sinensis and two freshwater species, Austropotamobius pallipes and Pacifastacus leniusculus. Exposure trials based upon natural (feeding) and artificial (intra-muscular injection) routes of exposure to WSSV revealed universal susceptibility to WSSV infection in these hosts. However, the relative degree of susceptibility (measured by progression of infection to disease, and mortality) varied significantly between host species. In some instances (Type 1 hosts), pathogenesis mimicked that observed in penaeid shrimp hosts whereas in other examples (Types 2 and 3 hosts), infection did not readily progress to disease, even though hosts were considered as infected and susceptible according to accepted principles. Results arising from challenge studies are discussed in relation to the potential risk posed to non-target hosts by the inadvertent introduction of WSSV to European waters via trade. Furthermore, we highlight the potential for

  9. New knowledge about 'white spots' in superalloys

    Jackman, L.A. (Teledyne Allvac, Monroe, N.C. (United States)); Maurer, G.E. (Special Metals Corp., New Hartford, N.Y. (United States)); Widge, S. (Carpenter Technology Corp., Reading, PA (United States))

    1993-05-01

    In April 1991, the first in a series of workshops was held to discuss ways in which the gas turbine industry could better understand defects in nickel-base superalloys. The group's primary objective was to better define, and expand knowledge about, segregation in superalloys such as Alloy 718 and Waspaloy,with emphasis on light-etching areas referred to as solute-lean defects or 'white spots'. This 'White Spots Committee' formed four subcommittees to focus efforts on classification, inspection, mechanisms, and mechanical properties. Completion of the tasks that these subcommittees have undertaken should greatly improve the gas turbine industry's understanding of the physical and mechanical nature of white spots. The primary purpose of this article is to formalize the characterization and classification of white spots in high-strength superalloys so that the metallurgical community can begin to use a common vocabulary when referring to them. An overview of formation mechanisms is presented along with a brief description of detection methods. Also discussed are preliminary test results, which should help shed light on the effects of solute-lean microstructures on tensile and fatigue properties. Although white spots are not limited to any single superalloy or class of superalloy, Alloy 718 is emphasized because it is so widely used, and because its relatively large solidus-liquidus temperature interval ([approximately]75 C, 135 F) and high niobium content ([approximately]5.3% Nb) make it prone to segregation. Three distinct types of white spots have been identified and named by the committee: discrete, dendritic, and solidification white spots.

  10. White spots in pharmaceutical pipelines-EMA identifies potential areas of unmet medical needs.

    Papaluca, Marisa; Greco, Martina; Tognana, Enrico; Ehmann, Falk; Saint-Raymond, Agnès

    2015-05-01

    Unmet medical needs are a priority for organizations such as the WHO and major public-private initiatives, such as Innovative Medicines Initiative, were established to speed up the development of better and safer medicines for patients. To assist such projects, the EMA in its 'Road Map to 2015' considered the mapping of unmet medical needs as a priority. This study has identified medical conditions for which the EMA could not identify developments in the pharmaceutical pipelines, that is, 'white spots'. Our analysis was made using external data sources as well as mining data of the EMA. The main areas for white spots were oncology, infectious diseases and certain psychiatric conditions. According to our data and a review of literature, in a number of these white spots, diagnostic tools may even be missing. The identification of those conditions will benefit stakeholders, including regulators, research funding bodies and patients' organizations.

  11. White spot formation under orthodontic bands cemented with glass ionomer with or without Fluor Protector.

    van der Linden, R P; Dermaut, L R

    1998-06-01

    The purpose of this study was to determine whether an additional application of Fluor Protector before band cementation with glass ionomer cement reduces white spot formation compared with band cementation with glass ionomer cement. In the in vitro study, 80 premolars were divided in half, creating a control and a test group. All specimens were divided into four different groups to simulate different clinical situations and stored in a demineralizing solution to induce white spot formation. In the in vivo investigation, 18 orthodontic patients were incorporated in the study. One lower and one upper first molar band (randomly selected) were coated with Fluor Protector and then cemented with a glass ionomer cement (test group). The other two uncoated first molars were cemented with glass ionomer cement and served as the control group. The application of Fluor Protector in combination with Aquacem did not contribute to a reduction of white spot formation underneath molar bands compared with the use of Aquacem for banding.

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

    Bröchner, Ann; Christensen, Carsten; Kristensen, Bjarne;

    2010-01-01

    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...... 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...

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

    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.

  14. A review on prevention and treatment of post-orthodontic white spot lesions - evidence-based methods and emerging technologies

    Bergstrand, Fredrik; Twetman, Svante

    2011-01-01

    The aim of this paper was to update the evidence for primary and secondary prevention (treatment) of white spot lesions (WSL) adjacent to fixed orthodontic appliances.......The aim of this paper was to update the evidence for primary and secondary prevention (treatment) of white spot lesions (WSL) adjacent to fixed orthodontic appliances....

  15. Efficacy of Ichthyophthirius vaccines in channel catfish against white spot disease

    Ichthyophthirius (Ich) is a protozoan that causes white spot disease in many cultured fish and lead to severe losses in aquaculture. Two trials were conducted to determine the efficacy and serum antibody response of different formulation of Ich vaccines in channel catfish. In trial I, catfish were i...

  16. Quantification of white spot lesions around orthodontic brackets with image analysis.

    Livas, C.; Kuijpers-Jagtman, A.M.; Bronkhorst, E.; Derks, A.; Katsaros, C.

    2008-01-01

    OBJECTIVE: To investigate the use of image analysis for diagnosis and quantification of artificial white spot lesions on digital photographs before and after removal of orthodontic brackets. MATERIALS AND METHODS: Enamel demineralization was artificially induced on the labial surface of 20 teeth bon

  17. Management of Post-orthodontic White Spot Lesions and Subsequent Enamel Discoloration with Two Microabrasion Techniques

    Arezoo Jahanbin

    2015-03-01

    Full Text Available Statement of the Problem: Demineralization of enamel adjacent to orthodontic appliances frequently occurs, commonly due to insufficient oral hygiene. Purpose: The aim of this study was to compare two microabrasion techniques on improving the white spot lesions as well as subsequent enamel discoloration. Materials and Method: Sixty extracted premolar teeth without caries and hypo-plasia were selected for this study. White spot lesions were artificially induced on the buccal surface of each tooth. Teeth were randomly assigned to three treatment groups, each treated with pumice powder as the control, microabrasion with 18% HCl, and microabrasion with 37% H3PO4. Subsequently, the three groups were daily immersed for five minutes in a tea-coffee solution for a period of one week. Colorimetric evaluation was done before and after formation of white spot lesions, after microabrasion, and after immersion in the colored solution; then the color differences (∆E were calculated. Statistical analysis was performed by multiple measurement analysis and the Tukey’s test. Results: This study showed that ∆E between the stages of white spot formation and microabrasion for H3PO4 was more than other groups and for the pumice powder group it was less than the others. Furthermore, there was a significant difference between ∆E of the three study groups (p= 0.017. Additionally, ∆E after placing the teeth in the colored solution and microabrasion was the highest for the HCl group and the lowest for the pumice powder group. There was also a significant difference between the three groups (p= 0.000. Conclusion: Pumice powder alone had similar effects as 18% HCl on removing the white spot lesions. Nevertheless, 18% HCl makes the enamel susceptible for subsequent color staining more than the other microabrasion methods.

  18. 南中国斑节对虾养殖中控制白斑综合症病的理论和策略%Theory and Strategies for Controlling White Spot Syndrome (WSS) of Cultured Penaeus monodon in South China

    2000-01-01

    @@ White spot syndrome virus (WSSV) is the causative agent of white spot syndrome (WSS) of cultured penaeid shrimps. WSS breaks out and prevails in cultured penaeid shrimps in many countries and regions of the world, especially in southeast Asia. WSSV is the virus most severe damaging to the cultured penaeid shrimps in the world. At the present time, the control of the outbreaks of WSSV will recover and develop the penaeid shrimp cultures in China and even in the world.

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

    de Jesús Durán-Avelar, Ma; 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.

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

    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.

  1. 环介导恒温扩增技术快速检测对虾白斑综合征病毒方法的建立%Establishment of Loop-mediated Isothermal Amplification for Detection of White spot syndrome virus

    李明云; 丁文超; 陈炯; 史雨红

    2011-01-01

    DNA环介导恒温扩增(loop-mediated isothermal amplification,LAMP)技术是一种特异、灵敏、快速的新型基因检测技术,其检测结果可以通过琼脂糖凝胶电泳观察,也可以通过加入核酸染料SYBR Green I进行肉眼观察.本研究根据对虾白斑综合征病毒(White spot syndrome virus,WSSV)的结构蛋白VP26基因序列,设计一套引物,成功建立了针对WSSV的LAMP检测方法.应用该方法,在65℃温育1h的条件下快速扩增病毒基因组DNA,琼脂糖凝胶电泳得到特异性梯度条带;在反应体系中添加SYBR Green I染料后,可通过肉眼观察有无荧光直接判定结果.该研究建立的LAMP法能特异性检出WSSV,其检测下限比PCR法低一个数量级,相当于0.563 pg/μL的质粒印MD19-T-VP26)浓度.表明所建立的WSSV LAMP检测法特异性好、灵敏度高、操作简单方便,有望作为WSSV快速诊断的常规方法.%Loop-mediated isothermal amplification (LAMP) method is a novel gene detection technique with specificity, sensitive and rapidity, the products of which can be electrophoresed in 1.5% agarose gels, or visually inspected by adding nucleic acid dye (SYBR Green Ⅰ ). In this paper, a one step loop-mediated isothermal amplification (LAMP) assay was developed for detection of White spot syndrome virus (WSSV). A set of primers were designed from the VP26 gene sequence of WSSV. The assay was established to amplify WSSV genomic DNA by incubation at 65℃ for only 1 h, and required only a simple water bath or heating block to provide a constant temperature of 65℃. LAMP amplification products had a ladder-like appearance when electrophoresed on an agarose gel. With the addition of SYBR Green Ⅰ, the presence of the target DNA could be detected by naked eyes. The detection limit of plasmid DNA (pMD19-T-VP26) by LAMP was 0.563 pg/μL which was found to be lower than that of the commonly used PCR method. The results indicate the suitability and simplicity of the test as

  2. Case Report:White-spot disease of Chinese soft-shelled turtles (Trionyx sinens) caused by Paecilomyces lilacinus

    Xiao-liang LI; Chu-long ZHANG; Wei-huan FANG; Fu-cheng LIN

    2008-01-01

    Chinese soft-shelled turtles (Trionyx sinens) in culture farms using an artificial warming system in Zhejiang, China,often show typical signs of white-spot disease such as white spots on their bodies, skin lesions, anorexia and eventually death. The sick turtles were mostly 5~80 g in weight. A suspected fungai pathogen was isolated from the sick turtles and verified as Pae-cilomyces lilacinus by sequence analysis of the internal transcribed spacer (ITS) of its ribosomal DNA (rDNA). Detailed mor-phological examinations were also conducted to confirm the white-spot disease.

  3. Effect of Fluoridated Toothpaste on White Spot Lesions in Postorthodontic Patients

    2013-01-01

    ABSTRACT Introduction: This article illustrates a new treatment approach and evaluates the effect of use of fluoridated toothpaste on the remineralization of white spot lesions (WSLs) diagnosed at debonding. Materials and methods: Thirty-one orthodontic patients (mean age: 19.6 years), with a minimum of four WSLs on the buccal surfaces of the maxillary and mandibular incisors, canines, premolars and first molars after orthodontic therapy, were enrolled in a double-blind, randomized, longitudi...

  4. Fluoride-Releasing Materials to Prevent White Spot Lesions around Orthodontic Brackets: A Systematic Review

    Nascimento,Patrícia Layane de Menezes Macêdo; Fernandes,Micaelle Tenório Guedes; de Figueiredo, Fabricio Eneas Diniz; Faria-e-Silva,André Luis

    2016-01-01

    Abstract The relation between orthodontic fixed appliances use and enamel demineralization is well established. Different preventive approaches have been suggested to this problem, but controversy remains about which is the best. The aim of this study was to perform a systematic review of clinical trials that investigated the effectiveness of materials containing fluorides to lute brackets or cover the bonding interface in order to inhibit the development and progression of white spot lesions...

  5. Fundus white spots and acquired night blindness due to vitamin A deficiency.

    Genead, Mohamed A; Fishman, Gerald A; Lindeman, Martin

    2009-12-01

    To report a successfully treated case of acquired night blindness associated with fundus white spots secondary to vitamin A deficiency. An ocular examination, electrophysiologic testing, as well as visual field and OCT examinations were obtained on a 61-year-old man with vitamin A deficiency who had previously undergone gastric bypass surgery. The patient had a re-evaluation after treatment with high doses of oral vitamin A. The patient was observed to have numerous white spots in the retina of each eye. Best-corrected visual acuity was initially 20/80 in each eye, which improved to 20/40-1 OU after oral vitamin A therapy for 2 months. Full field electroretinogram (ERG) testing, showed non-detectable rod function and a 34 and 41% reduction for 32-Hz flicker and single flash cone responses, respectively, below the lower limits of normal. Both rod and cone functions markedly improved after initiation of vitamin A therapy. Vitamin A deficiency needs to be considered in a patient with white spots of the retina in the presence of poor night vision.

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

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

    2016-01-01

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

  7. cDNA微阵列结合抑制性差减杂交研究对虾白斑综合征病毒部分表达基因%Analysis of Partial Expressed Genes of Penaeid Shrimp White Spot Syndrome Virus with Suppression Subtractive Hybridization and cDNA Microarray

    曾勇; 陆承平

    2004-01-01

    通过抑制性差减杂交技术建立了包含对虾白斑综合征病毒表达性基因的差减文库,并用cDNA微阵列技术进行了鉴定,得到255个正向克隆.对其中的184个正向阳性克隆进行了测序,测序结果通过BLAST与GenBank中序列进行比对,共得到WSSV(white spot syndrome virus,WSSV)基因30个.此次首次鉴定了5个,其中WSV184具有调控蛋白的结构特征(Cys2/Cys2型锌指),WSV321和WSV322含跨膜结构,且存在可能的糖基化位点.有3个被其它研究推断无polyA结构的阅读框所处的mRNA应有polyA结构.进一步用Dot Northern blot对克隆号PCI118(含WSSV阅读框WSV321和WSV322)进行鉴定,表明确实存在该基因的转录.进而根据已报道的WSSV基因序列设计两条引物,用快速扩增cDNA末端技术,扩增WSV321和WSV322两个阅读框所处的cD-NA的5′端片段和3′端片段,分析得到其全长共1 109bp,与已报道的WSSV全基因序列(AF332093)的相关序列完全相同.该mRNA存在polyA,并有加尾信号AATAAA;两个阅读框都没有自己的TATA盒,但都有病毒RNA聚合酶Ⅱ的结合位点-CCAAT盒;它们编码的蛋白质分别有117和227个氨基酸,都存在可能的糖基化位点,其中WSV321一个,WSV322两个.

  8. Flow fields within Jupiter's Great Red Spot and White Oval BC

    Mitchell, J. L.; Garneau, G. W.; Beebe, R. F.; Ingersoll, A. P.

    1981-01-01

    Voyager 1 high-resolution images of Jupiter's Great Red Spot (GRS) and White Oval BC are used to map flow fields within these two areas. The relative vorticity is computed as a function of semi-major axis length and position angle in a coordinate system consisting of concentric ellipses of equal eccentricity. Wind speeds of 110-120 m/s are observed near the outer edge of both features, and along their minor axes relative vorticity profiles reach a maximum of 0.00006/s. Maximum Rossby numbers of 0.36 are computed for flows within both features, and are found to be low, indicating geostrophic constraints on the flow. The difference in streamline curvature within the GRS and the Oval BC is found to compensate for the difference in planetary vorticity at the respective latitudes of the features. Finally, motions within the central region of the GRS are slower and more random than around the spot's outer portion.

  9. First Report of the Natural Occurrence of Tomato chlorotic spot virus in Peanuts in Haiti

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

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

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

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

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

    2007-01-01

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

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

    Geerts-Dimitriadou, C.; Lu, Y.Y.; Geertsema, C.; Goldbach, R.W.; Kormelink, R.J.M.

    2012-01-01

    The intergenic region (IR) of ambisense RNA segments from animal- and plant-infecting (-)RNA viruses functions as a bidirectional transcription terminator. The IR sequence of the Tomato spotted wilt virus (TSWV) ambisense S RNA contains stretches that are highly rich in A-residues and U-residues and

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

    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,

  14. IMMUNE-RELATED GENES EXPRESSION AND PHAGOCYTOSIS AGAINST WHITE SPOT SYNDROME VIRUS AFTER ORAL DELIVERY OF VP28 USING BACILLUS SUBTILIS AS VEHICLES IN LITOPENAEUS VANNAMEI%以枯草芽孢杆菌递呈VP28对南美白对虾免疫相关基因表达和细胞特异性吞噬的影响

    丁晶; 王彦波; 傅玲琳

    2013-01-01

    以枯草芽孢杆菌(Bacillus subtilis)为活载体口服递呈对虾白斑综合征病毒(WSSV)囊膜蛋白 VP28,评价其抗病毒感染能力、对南美白对虾免疫相关基因表达以及血淋巴细胞对病毒特异性吞噬的影响。经口服免疫枯草重组菌株B. subtilis-VP28攻毒后,对虾的相对存活率达83.3%。为探讨重组菌株的抗病机理,比较研究了免疫相关基因-proPO(酚氧化酶原)、Peroxinectin(PE)和脂多糖-β-1,3-葡聚糖结合蛋白(LGBP)基因的表达差异,并进一步分析了血淋巴细胞吞噬活性和特异性。结果表明, B. subtilis-VP28菌液能显著提高(P<0.05)对虾proPO、PE和LGBP mRNA的表达水平和血细胞对WSSV的吞噬活性, B. subtilis组对免疫相关基因也有一定的激活作用,而B. subtilis-VP28发酵上清液则能增加血细胞吞噬活性;此外, B. subtilis-VP28菌液组血细胞对WSSV具有特异性吞噬作用。研究为枯草重组菌株B. subtilis-VP28抗WSSV感染作用及其作为特殊功能水产微生态制剂的应用提供了一定的科学依据。%The regulation of immune-related genes expression and phagocytosis of White Spot Syndrome Virus (WSSV) were evaluated by oral delivery of VP28 using Bacillus subtilis as vehicles in Litopenaeus vannamei. In our initial ex-periment, by oral delivery of B. subtilis spores harboring VP28 (B. subtilis-VP28) to L. vannamei, the extremely high survival (Relative Percent Survival:83.3%) upon challenge with WSSV can be observed. The differences of genes ex-pression levels of proPO, Peroxinectin (PE) and lipopolysaccharide-and beta-1, 3-glucan-binding protein (LGBP) were demonstrated among experimental groups of B. subtilis-VP28 bacterial spores, B. subtilis-VP28 supernatants, B. subtilis and control. The result showed that immune-related genes (proPO, PE and LGBP) were significantly (P<0.05) upregu-lated in both B. subtilis-VP28 bacterial spores and B. subtilis feeding groups compared to B. subtilis-VP28

  15. Bonding brackets on white spot lesions pretreated by means of two methods

    Vianna, Julia Sotero; Marquezan, Mariana; Lau, Thiago Chon Leon; Sant'Anna, Eduardo Franzotti

    2016-01-01

    Abstract Objective: The aim of this study was to evaluate the shear bond strength (SBS) of brackets bonded to demineralized enamel pretreated with low viscosity Icon Infiltrant resin (DMG) and glass ionomer cement (Clinpro XT Varnish, 3M Unitek) with and without aging. Methods: A total of 75 bovine enamel specimens were allocated into five groups (n = 15). Group 1 was the control group in which the enamel surface was not demineralized. In the other four groups, the surfaces were submitted to cariogenic challenge and white spot lesions were treated. Groups 2 and 3 were treated with Icon Infiltrant resin; Groups 4 and 5, with Clinpro XT Varnish. After treatment, Groups 3 and 5 were artificially aged. Brackets were bonded with Transbond XT adhesive system and SBS was evaluated by means of a universal testing machine. Statistical analysis was performed by one-way analysis of variance followed by Tukey post-hoc test. Results: All groups tested presented shear bond strengths similar to or higher than the control group. Specimens of Group 4 had significantly higher shear bond strength values (p < 0.05) than the others. Conclusion: Pretreatment of white spot lesions, with or without aging, did not decrease the SBS of brackets. PMID:27275613

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

    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.

  17. Effect of acid-etching on remineralization of enamel white spot lesions.

    Al-Khateeb, S; Exterkate, R; Angmar-Månsson, B; ten Cate, J M; ten Cate, B

    2000-02-01

    This in vitro study aimed at investigating whether full remineralization would occur in white spot lesions when the surface porosity was increased by acid-etching. The effect of fluoride was also investigated. Enamel blocks with in vitro produced white spot lesions were used. Group A was exposed to a remineralizing solution only. In group B, the lesions were etched with 35% phosphoric acid for 30 s, then treated as in group A. Group C was treated as group A + daily treatment with a fluoride toothpaste slurry (1,000 ppm) for 5 min. Group D was treated as group B + the daily fluoride treatment of group C. The remineralization was measured weekly with Quantitative Light-induced Fluorescence during the experimental period. After 10 weeks of remineralization, mineral profiles were assessed with transverse microradiography. The enamel fluorescence was partly regained. There were significant differences in the lesion depth, mineral content at the surface layer, and integrated mineral loss between the groups. Addition of fluoride accelerated the remineralization only in the beginning; in later stages the process leveled out and even reached a plateau in all the groups. It was concluded that full remineralization was not achieved by etching, by the addition of fluoride, nor by the combination of both treatments in this in vitro study.

  18. Bonding brackets on white spot lesions pretreated by means of two methods

    Julia Sotero Vianna

    2016-04-01

    Full Text Available Abstract Objective: The aim of this study was to evaluate the shear bond strength (SBS of brackets bonded to demineralized enamel pretreated with low viscosity Icon Infiltrant resin (DMG and glass ionomer cement (Clinpro XT Varnish, 3M Unitek with and without aging. Methods: A total of 75 bovine enamel specimens were allocated into five groups (n = 15. Group 1 was the control group in which the enamel surface was not demineralized. In the other four groups, the surfaces were submitted to cariogenic challenge and white spot lesions were treated. Groups 2 and 3 were treated with Icon Infiltrant resin; Groups 4 and 5, with Clinpro XT Varnish. After treatment, Groups 3 and 5 were artificially aged. Brackets were bonded with Transbond XT adhesive system and SBS was evaluated by means of a universal testing machine. Statistical analysis was performed by one-way analysis of variance followed by Tukey post-hoc test. Results: All groups tested presented shear bond strengths similar to or higher than the control group. Specimens of Group 4 had significantly higher shear bond strength values (p < 0.05 than the others. Conclusion: Pretreatment of white spot lesions, with or without aging, did not decrease the SBS of brackets.

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

    Bröchner, Ann; Christensen, Carsten; Kristensen, Bjarne; Tranæus, Sofia; Karlsson, Lena; Sonnesen, Liselotte; Twetman, Svante

    2011-06-01

    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 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.

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

    Paiva, Lilian Fernanda Santos; Fidalgo, Tatiana Kelly da Silva; Maia, Lucianne Cople

    2014-01-01

    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-ionomers 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 white spot lesion formation was evaluated macroscopically (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 white spot formation around restorations in comparison with Ionomaster R™ (p white spot formation and presented a preventive effect (p = 0.051). GICs presented a greater percentage of fluoride, aluminum, and strontium, and proved effective in white spot lesion prevention around restorations.

  1. The hemic response of white-spotted bamboo sharks (Chiloscyllium plagiosum) with inflammatory disease.

    Alexander, Amy B; Parkinson, Lily A; Grant, Krystan R; Carlson, Eric; Campbell, Terry W

    2016-05-01

    As elasmobranch medicine becomes more commonplace, there continues to be confusion with techniques and evaluation of the shark hemogram and it remains unknown if they are able to mount an inflammatory hemic response. The aims of this study were to compare two total white blood cell (WBC) count techniques, establish a reference interval for captive white-spotted bamboo sharks (Chiloscyllium plagiosum), and determine if elasmobranchs are capable of mounting an inflammatory hemic response. Correlation statistics were performed on hematologic results for healthy female bamboo sharks to assess the use of Natt-Herrick's and phloxine methods. Total WBC counts and differentials were obtained from males with severe traumatic clasper wounds and compared to the healthy females. We elected clasper amputation as the preferred treatment intervention and post-operative hematology was performed one month later. There was poor correlation of leukocyte counts between the two WBC count methods. Hematologic values were established for the females and males pre- and post-operatively. Males with wounds had a marked leukocytosis and heterophilia. Post-operative blood work showed a resolution of total WBC count and a trend toward resolution of the heterophilia. This study provides hematologic values for white-spotted bamboo sharks and confirms that the Natt-Herrick's method is preferred for lymphocytic species. Hematologic differences present in males with clasper wounds suggests that elasmobranchs do mount an inflammatory hemic response. Treatment via clasper amputation proved to be a safe and efficient means for clinical treatment that led to a trend toward resolution of the inflammatory leukogram. Zoo Biol. 35:251-259, 2016. © 2016 Wiley Periodicals, Inc.

  2. Genetic Dissection of Novel QTLs for Resistance to Leaf Spots and Tomato Spotted Wilt Virus in Peanut (Arachis hypogaea L.)

    Pandey, Manish K.; Wang, Hui; Khera, Pawan; Vishwakarma, Manish K.; Kale, Sandip M.; Culbreath, Albert K.; Holbrook, C. Corley; Wang, Xingjun; Varshney, Rajeev K.; Guo, Baozhu

    2017-01-01

    Peanut is an important crop, economically and nutritiously, but high production cost is a serious challenge to peanut farmers as exemplified by chemical spray to control foliar diseases such as leaf spots and thrips, the vectors of tomato spotted wilt virus (TSWV). The objective of this research was to map the quantitative trait loci (QTLs) for resistance to leaf spots and TSWV in one recombinant inbred line (RIL) mapping population of “Tifrunner × GT-C20” for identification of linked markers for marker-assisted breeding. Here, we report the improved genetic linkage map with 418 marker loci with a marker density of 5.3 cM/loci and QTLs associated with multi-year (2010–2013) field phenotypes of foliar disease traits, including early leaf spot (ELS), late leaf spot (LLS), and TSWV. A total of 42 QTLs were identified with phenotypic variation explained (PVE) from 6.36 to 15.6%. There were nine QTLs for resistance to ELS, 22 QTLs for LLS, and 11 QTLs for TSWV, including six, five, and one major QTLs with PVE higher than 10% for resistance to each disease, respectively. Of the total 42 QTLs, 34 were mapped on the A sub-genome and eight mapped on the B sub-genome suggesting that the A sub-genome harbors more resistance genes than the B sub-genome. This genetic linkage map was also compared with two diploid peanut physical maps, and the overall co-linearity was 48.4% with an average co-linearity of 51.7% for the A sub-genome and 46.4% for the B sub-genome. The identified QTLs associated markers and potential candidate genes will be studied further for possible application in molecular breeding in peanut genetic improvement for disease resistance. PMID:28197153

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

    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.

  4. Prevention and Treatment of White Spot Lesions During and After Treatment with Fixed Orthodontic Appliances: a Systematic Literature Review

    Kristina Lopatiene

    2016-06-01

    Full Text Available Objectives: The aim of the systematic literature review is to update the evidence for the prevention of white spot lesions, using materials containing fluoride and/or casein phosphopeptide-amorphous calcium phosphate during and after treatment with fixed orthodontic appliances. Material and Methods: Information search for controlled studies on humans published between January 2008 and February 2016 was performed in PubMed, ScienceDirect, Embase, The Cochrane Library. Inclusion criteria were: the English language, study on humans, patients undergoing orthodontic treatment with fixed appliances, randomized or quasi-randomized controlled clinical studies fluoride-containing product or casein derivates used throughout the appliance therapy or straightaway after debonding. Results: 326 articles were reviewed (Embase 141, PubMed 129, ScienceDirect 41, Cochrane 15. Twelve clinical studies fulfilled all inclusion criteria. Use of fluoridated toothpaste had a remineralizing effect on white spot lesions (WSLs (P < 0.05; fluoride varnish and casein supplements were effective in prevention and early treatment of WSLs (P < 0.05. Conclusions: Early detection of white spot lesions during orthodontic treatment would allow implementing preventive measures to control the demineralization process before lesions progress. The systemic review has showed that the usage of fluoride and casein supplements in ameliorating white spot lesions during and after fixed orthodontic treatment is significantly effective. However the use of casein phosphopeptide-amorphous calcium phosphate can be more beneficial than fluoride rinse in the reduction of demineralization spots.

  5. Four kinds of ENU-induced white spot mice and chromosome locations of the mutant genes

    WU Baojin; MAO Huihua; SHAO Yixiang; XUE Zhengfeng; LI Houda

    2003-01-01

    Phenotype-driven is the name for an approach used to study gene functions through mutagenesis, location and cloning of the mutant gene. In this study, 150 male C57BL/6J(B6) mice were treated with ENU and reproduced a total offspring of 3860. Of these descendants, 210 exhibited mutation phenotypes by screening, and more than 10 of them are hereditable. Four kinds of mutant mice, named Wbct, W-1Bao, W-2Bao, and W-3Bao, showed dominant hereditary white spot mutation with partial albinism on their belly, distal limbs and tail terminal. To map these mutant genes, 39 microsatellites, equally distributed on the mouse genome and with difference between B6 and DBA/2J (D2), were selected to scan the genome after discrimination of the white spots in the F2 mice [(B6×D2)×D2]. It is found that, the log odds score (LODS) between W-1Bao and D5Mit168 is 0.56, and the LODS of W-1Bao and D5Mit352 is 4.47. With the gradual application of microsatellites D5Mit290, D5Mit312, D5Mit308 and D5Mit356 that are close to the mutant gene, and the number of F2 mice going up to 537, the mutant W-1Bao is located between D5Mit356 and D5Mit308 on chromosome 5, about 42.19 cM from the centromere. In the same way, W-2Bao and W-3Bao are mapped nearby W-1Bao, and Wbct is located on chromosome 1, about 41.6 cM from the centromere. After searching for the mouse genome database (MGD) and performing a one-by-one study of all genes located on chromosome subregion, it is believed that the kit gene is an excellent candidate for the white spot mutations of W-1Bao, W-2Bao and W-3Bao.

  6. Prevalence of white-spot lesions before and during orthodontic treatment with fixed appliances.

    Lucchese, Alessandra; Gherlone, Enrico

    2013-10-01

    The aim of the study was to determine the prevalence of white-spot lesions (WSLs) in patients with fixed orthodontic appliances. The cross-sectional study sample consisted of three groups of patients: group I, 59 patients treated orthodontically for 6 months; group II, 64 patients treated for 12 months; group 0 (control), 68 patients examined immediately before appliance placement. All groups were treated with a 0.022-inch slot preadjusted appliance and they wore a functional fixed appliance. The presence of WSLs was evaluated by visual examination using the scoring system proposed by Gorelick. The groups were evaluated for differences in the prevalence of at least one WSL using Fisher's exact test, followed by Bonferroni pairwise comparisons. The prevalence of WSLs by tooth type was evaluated with logistic regression (P orthodontic bonding. Considering how quickly these lesions can develop and become irreversible, early diagnosis is of critical importance.

  7. Measuring color change of tooth enamel by in vitro remineralization of white spot lesion.

    Betina Tolcachir

    2015-12-01

    Full Text Available Objective colour determination is based on calculating the colorimetric distance (ΔE within a colour space. So far, the most used colour space in dentistry is CIE L*a*b (Comission Internationale de l´Éclairage. CIE L*C*h* has been recently developed, showing a better correlation with the perception of the human eye. Objective: To determine the ability of an in vitro remineralisation substance to blend the colour of white spot lesions (WSL with sound enamel, determining ΔE by using the CIE L*C*h* colour space. Methods: In vitro WSL was generated by immersing 10 samples obtained from human third molars in a demineralization solution for 72h. Amorphous calcium phosphate stabilized by casein phosphopeptide (CPP-ACP was then applied for 60 days while maintaining the samples in artificial saliva at 37ºC. To evaluate the colour of enamel, images were taken from the samples placed in specifically designed silicone moulds after generating the WSL (pre-stage and after remineralisation by scanning, applying the colorimetric distance equation (ΔE*CMC according to the Colour Measurement Committee. Results: Treatment with CPP-ACP caused a significant ΔE decrease with respect to the pre-stage (p<0.001, while the analysis of parameters that make up the colour showed a reduction in the difference of hue (∆H (p<0.001 and brightness (∆L (p<0.01 after applying CPP-ACP. Discussion: CPP-ACP penetrated to the depth of the white spot lesion, making its appearance similar to that of the sound enamel, probably because of the formation of different mineral phases than that of the original structure, although pores were not completely filled.

  8. Current technology and techniques in re-mineralization of white spot lesions: A systematic review

    Podray, Susan S.

    White Spot lesions are a common iatrogenic occurrence on patients who are treated with fixed orthodontic appliances. There is a dynamic chemical interaction between enamel and saliva at the tooth surface that allow a lesion to have phase changes involving demineralization of enamel and reminerlization. This is due to calcium and phosphate dissolved in saliva that is deposited onto the tooth surface or removed depending on the surrounding pH. Caseinphosphopeptide-amorphous calcium phosphate (CPP-ACP) is gaining popularity in dentistry as a way to increase the available level of calcium and phosphate in plaque and saliva to improve the chemical gradient so that if favors reminerlization. The aim of our investigation is to search the available current literature and formulate a recommendation for use of CPP-ACP in orthodontics. Publications from the following electronic databases were searched: PubMed, Web of Science, Cochrane Library and Science Direct. Searches from August 2010 to April 1st 2012 were performed under the terms "MI Paste OR Recaldent OR caseinphosphopeptide-amorphous calcium phosphate OR CPP-ACP or tooth mousse". The searches yielded 155 articles, These were reviewed for relevance based on inclusion and exclusion criteria. Articles with inappropriate study design or no outcome measures at both baseline and end point were also excluded. 13 articles were deemed of relevance with a high quality study design and were included in this study for evaluation. The current literature suggests a preventative treatment regimen in which MI Paste Plus is used. It should be delivered once daily prior to bed after oral hygiene for 3 minutes in a fluoride tray, throughout orthodontic treatment. It should be recommended for high risk patients determined by poor oral hygiene, as seen by the inability to remove plaque from teeth and appliances. This protocol may prevent or assist in the remineralization of enamel white spot lesions during and after orthodontic treatment.

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

    Maris, P.C.

    2004-01-01

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

  10. First report of Pelargonium zonate spot virus from tomato in the United States

    Pelargonium zonate spot virus (PZSV) was first isolated from tomato in southern Italy in 1982, and later was also reported from Spain and France. Infected tomato plants showed stunting, malformation, yellow rings and line patterns on the leaves, and concentric chlorotic ringspots on the stems. In Ju...

  11. In vitro transcription of Tomato spotted wilt virus is independent of translation

    Knippenberg, van I.C.; Goldbach, R.W.; Kormelink, R.J.M.

    2004-01-01

    Ongoing transcription in vitro of Tomato spotted wilt virus (TSWV) has previously been demonstrated to require the presence of reticulocyte lysate. This dependence was further investigated by testing the occurrence of transcription in the presence of two translation inhibitors: edeine, an inhibitor

  12. Two spinefoot colour morphs: mottled spinefoot Siganus fuscescens and white-spotted spinefoot Siganus canaliculatus are synonyms.

    Hsu, T-H; Adiputra, Y T; Burridge, C P; Gwo, J-C

    2011-11-01

    Mottled spinefoot Siganus fuscescens and white-spotted spinefoot Siganus canaliculatus are two similar species that differ subtly in colouration and morphology. Three major mtDNA clades were identified for these species, but individuals were clustered by amplified fragment length polymorphism (AFLP) according to geography rather than morphology, suggesting that the colour morphs are interbreeding.

  13. REGRESSION OF WHITE SPOT ENAMEL LESIONS - A NEW OPTICAL METHOD FOR QUANTITATIVE LONGITUDINAL EVALUATION IN-VIVO

    OGAARD, B; TENBOSCH, JJ

    1994-01-01

    This article describes a new nondestructive optical method for evaluation of lesion regression in vivo. White spot caries lesions were induced with orthodontic bands in two vital premolars of seven patients. The teeth were banded for 4 weeks with special orthodontic bands that allowed plaque accumul

  14. Esthetic improvement of white spot lesions and non-pitted fluorosis using resin infiltration technique: Series of four clinical cases

    Neeraj Gugnani

    2014-01-01

    Full Text Available White opacities have always been a major concern of esthetics for patients and can have varying etiology. In general, white discolorations of enamel might be due to dental fluorosis, early caries (white spot lesions, developmental defects etc. Conventional treatment options available for such opacities include non-invasive and invasive approaches. Recently, a new "micro-invasive" technique has been introduced as an alternative therapeutic approach that improves such opacities esthetically, in a single sitting, is painless and exhibits no complications. This case series illustrates the use of resin infiltration to treat fluorosis stains and WSLs, exhibiting significant improvement in esthetics.

  15. Characterization of a detective form of tomato spotted wilt virus

    Verkleij, F.N.

    1982-01-01

    The work described in this thesis was aimed at the elucidation of the nature of a defective form of TSWV which does not form complete particles during infection.Properties of TSWV and the existence of a defective form of this virus are described in Chapter 1. A survey of the literature on three diff

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

    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...

  17. 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

    2012-01-01

    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 seve...

  18. A Frameshift Mutation in KIT is Associated with  White Spotting in the Arabian Camel

    Heather Holl

    2017-03-01

    Full Text Available While the typical Arabian camel is characterized by a single colored coat, there are rare populations with white spotting patterns. White spotting coat patterns are found in virtually all domesticated species, but are rare in wild species. Theories suggest that white spotting is linked to the domestication process, and is occasionally associated with health disorders. Though mutations have been found in a diverse array of species, fewer than 30 genes have been associated with spotting patterns, thus providing a key set of candidate genes for the Arabian camel. We obtained 26 spotted camels and 24 solid controls for candidate gene analysis. One spotted and eight solid camels were whole genome sequenced as part of a separate project. The spotted camel was heterozygous for a frameshift deletion in KIT (c.1842delG, named KITW1 for White spotting 1, whereas all other camels were wild‐type (KIT+/KIT+. No additional mutations unique to the spotted camel were detected in the EDNRB, EDN3, SOX10, KITLG, PDGFRA, MITF, and PAX3 candidate white spotting genes. Sanger sequencing of the study population identified an additional five kITW1/KIT+ spotted camels. The frameshift results in a premature stop codon five amino acids downstream, thus terminating KIT at the tyrosine kinase domain. An additional 13 spotted camels tested KIT+/KIT+, but due to phenotypic differences when compared to the KITW1/KIT+ camels, they likely represent an independent mutation. Our study suggests that there are at least two causes of white spotting in the Arabian camel, the newly described KITW1 allele and an uncharacterized mutation.

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

    Anna Rudzińska-Langwald

    2014-01-01

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

  20. In vitro transcription of Tomato spotted wilt virus is independent of translation

    Knippenberg, van, B.M.; Goldbach, R.W.; Kormelink, R.J.M.

    2004-01-01

    Ongoing transcription in vitro of Tomato spotted wilt virus (TSWV) has previously been demonstrated to require the presence of reticulocyte lysate. This dependence was further investigated by testing the occurrence of transcription in the presence of two translation inhibitors: edeine, an inhibitor that still allows scanning of nascent mRNAs by the 40S ribosomal subunit, and cycloheximide, an inhibitor that completely blocks translation including ribosome scanning. Neither of these inhibitors...

  1. Potential vorticity and layer thickness variations in the flow around Jupiter's Great Red Spot and White Oval BC

    Dowling, Timothy E.; Ingersoll, Andrew P.

    1988-01-01

    Using Voyager images, layer thickness variations in the flow around Jupiter's Great Red Spot (GRS) and White Oval BC were investigated by treating potential vorticity as a conserved tracer. Fluid trajectories around the GRS and the White Oval BC were calculated assuming the flow to be frictionless, adiabatic, hydrostatic, and steady in the reference frame of the vortex. The data obtained constitute a useful diagnostic which will help to differentiate between models of Jovian vortices. Implications of the observations were studied in the context of a one-layer quasi-geostrophic model in which a thin upper weather layer, which contains the vortex, is supported hydrostatically by a much deeper lower layer.

  2. Pepper necrotic spot virus, a new tospovirus infecting solanaceous crops in Peru.

    Torres, Roger; Larenas, Javiera; Fribourg, Cesar; Romero, Javier

    2012-04-01

    Two virus isolates, T1 and T2, causing necrotic spots on leaves and stems of pepper and tomato, respectively, were isolated in the La Joya valley, Arequipa, Peru, in 2010. These two isolates were inoculated to differential hosts for tospoviruses and showed differential fitness: T1 induced necrotic local lesions in Vigna unguiculata, whereas T2 produced only chlorotic spots. The complete nucleotide sequence of the small (S) RNA from T2 and 1863 bp of the S RNA from T1 were determined. The deduced N protein sequence showed high amino acid identity (97%) between the isolates, indicating that the T1 and T2 are isolates of the same virus. Sequence comparisons indicated that the amino acid sequence of the N protein shared 53.49-87.98% identity with known American tospoviruses. Phylogenetic analysis of both the NSs and N proteins revealed that this new tospovirus belongs to the American group. We conclude that this tospovirus should be considered a member of a new species. The name Pepper necrotic spot virus (PNSV) is proposed.

  3. Deproteinization of tooth enamel surfaces to prevent white spot lesions and bracket bond failure: A revolution in orthodontic bonding

    Roberto Justus

    2016-01-01

    Orthodontic treatment success is jeopardized by the risk of development of white spot lesions (WSLs) around orthodontic brackets. Unfortunately, the formation of WSLs still remains a common complication during treatment in patients with poor oral hygiene. Nearly 75% of orthodontic patients are reported to develop enamel decalcification because of prolonged plaque retention around brackets. It is the orthodontist′s responsibility to minimize the risk of patients having enamel decalcifications ...

  4. Prevention and Treatment of White Spot Lesions During and After Treatment with Fixed Orthodontic Appliances: a Systematic Literature Review

    Kristina Lopatiene; Marija Borisovaite; Egle Lapenaite

    2016-01-01

    ABSTRACT Objectives The aim of the systematic literature review is to update the evidence for the prevention of white spot lesions, using materials containing fluoride and/or casein phosphopeptide-amorphous calcium phosphate during and after treatment with fixed orthodontic appliances. Material and Methods Information search for controlled studies on humans published between January 2008 and February 2016 was performed in PubMed, ScienceDirect, Embase, The Cochrane Library. Inclusion criteria...

  5. Enhancement of superoxide dismutase and catalase activity in juvenile brown shrimp, Farfantepenaeus californiensis (Holmes, 1900, fed β-1.3 glucan vitamin E, and β-carotene and infected with white spot syndrome virus Incremento de la actividad superóxido dismutasa y catalasa en juveniles de camarón café Farfantepenaeus californiensis (Holmes, 1900 alimentados con β-1,3 glucano vitamina E y β-caroteno e infectados con el virus de la mancha blanca

    Rosario Pacheco

    2011-11-01

    Full Text Available The effect of dietary β-Ο-glucan, vitamin E, and β-carotene supplements in juvenile brown shrimp, Farfantepenaeus californiensis, inoculated with white spot syndrome virus (WSSV was evaluated. Groups of 30 organisms (weighing 1 ± 0.5 g were cultured in 60 L fiberglass tanks and fed daily with β-1.3-glucan (0.1%, vitamin E (0.01%, and β-carotene (0.01% for 23 days; the specimens were then inoculated with WSSV. The antioxidant activity of the enzymes superoxide dismutase (SOD and catalase (CAT were determined in the hepatopancreas and muscle at 0, 1, 6, 12, 24, and 48 h after inoculation. Shrimp fed with β-1.3-glucan, vitamin E, and β-carotene significantly increased SOD activity in the hepatopancreas and muscle at 12 and 24 h post-infection, respectively. Shrimp fed with vitamin E and β-1.3-glucan registered an increment in SOD activity from 12 to 48 h post-infection. Shrimp fed with β-carotene increased SOD activity before infection with WSSV, and shrimp fed with β-1.3-glucan and vitamin E increased CAT activity, also before infection. The CAT activity response in shrimp muscle increased with respect to the control group for all treatments tested from 1 to 6 h after inoculation with WSSV. The highest antioxidant response was registered in shrimp fed with vitamin E. Juvenile shrimp fed with vitamin E and later inoculated with WSSV registered 100% mortality at 72 h, but shrimp fed with β-Ο-glucan and β-carotene showed greater resistance to WSSV, with mortality at 144 h post-infection. This study demonstrated the capacity of juvenile Farfantepenaeus californiensis fed β-Ο-glucan, vitamin E, or β-carotene to increase the antioxidant response before and after viral infection.Se evaluó el efecto de β-1,3-glucano, vitamina E y β-caroteno en la dieta de juveniles de camarón café Farfantepenaeus californiensis inoculados con virus del síndrome de la mancha blanca (WSSV. Se colocaron grupos de 30 camarones (peso 1 ± 0,5 g en

  6. Dynamics of Saturn’s 2010 Great White Spot from high-resolution Cassini ISS observations

    Hueso, Ricardo; Sánchez-Lavega, A.; del Río-Gaztelurrutia, T.

    2012-10-01

    On December 5th 2010 a storm erupted in Saturn’s North Temperate latitudes which were experiencing early spring season. The storm quickly developed to a planet-wide disturbance of the Great White Spot type. The ISS instrument onboard Cassini acquired its first images of the storm on 23th December 2010 and performed repeated observations with a variety of spatial resolutions over the nearly 10 months period the storm continued active. Here we present an analysis of two of the image sequences with better spatial resolution of the mature storm when it was fully developed and very active. We used an image correlation algorithm to measure the cloud motions obtained from images separated 20 minutes and obtained 16,000 wind tracers in a domain of 60 degrees longitude per 20 degrees in latitude. Intense zonal and meridional motions accompanied the storm and reached values of 120 m/s in particular regions of the active storm. The storm released a chain of anticyclonic and cyclonic vortices at planetocentric latitudes of 36° and 32° respectively. The short time difference between the images results in estimated wind uncertainties of 15 m/s that did not allow to perform a complete analysis of the turbulence and kinetic spectrum of the motions. We identify locations of the updrafts and link those with the morphology in different observing filters. The global behaviour of the storm was examined in images separated by 10 hours confirming the intensity of the winds and the global behaviour of the vortices. Acknowledgments: This work was supported by the Spanish MICIIN project AYA2009-10701 with FEDER funds, by Grupos Gobierno Vasco IT-464-07 and by Universidad País Vasco UPV/EHU through program UFI11/55.

  7. Morphology and mechanics of the teeth and jaws of white-spotted bamboo sharks (Chiloscyllium plagiosum).

    Ramsay, Jason B; Wilga, Cheryl D

    2007-08-01

    The teeth of white-spotted bamboo sharks (Chiloscyllium plagiosum) are used to clutch soft-bodied prey and crush hard prey; however, the dual function is not evident from tooth morphology alone. Teeth exhibit characteristics that are in agreement with a clutching-type tooth morphology that is well suited for grasping and holding soft-bodied prey, but not for crushing hard prey. The dual role of this single tooth morphology is facilitated by features of the dental ligament and jaw joint. Tooth attachment is flexible and elastic, allowing movement in both sagittal and frontal planes. During prey capture spike-like tooth cusps pierce the flesh of soft prey, thereby preventing escape. When processing prey harder than the teeth can pierce the teeth passively depress, rotating inward towards the oral cavity such that the broader labial faces of the teeth are nearly parallel to the surface of the jaws and form a crushing surface. Movement into the depressed position increases the tooth surface area contacting prey and decreases the total stress applied to the tooth, thereby decreasing the risk of structural failure. This action is aided by a jaw joint that is ventrally offset from the occlusal planes of the jaws. The offset joint position allows many teeth to contact prey simultaneously and orients force vectors at contact points between the jaws and prey in a manner that shears or rolls prey between the jaws during a bite, thus, aiding in processing while reducing forward slip of hard prey from the mouth. Together the teeth, dental ligament, and jaws form an integrated system that may be beneficial to the feeding ecology of C. plagiosum, allowing for a diet that includes prey of varying hardness and elusiveness.

  8. Molecular variability analyses of Apple chlorotic leaf spot virus capsid protein

    T Rana; V Chandel; Y Kumar; R Ram; V Hallan; A A Zaidi

    2010-12-01

    The complete sequences of the coat protein (CP) gene of 26 isolates of Apple chlorotic leaf spot virus (ACLSV) from India were determined. The isolates were obtained from various pome (apple, pear and quince) and stone (plum, peach, apricot, almond and wild Himalayan cherry) fruit trees. Other previously characterized ACLSV isolates and Trichoviruses were used for comparative analysis. Indian ACLSV isolates among themselves and with isolates from elsewhere in the world shared 91–100% and 70–98% sequence identities at the amino acid and nucleotide levels, respectively. The highest degree of variability was observed in the middle portion with 9 amino acid substitutions in contrast to the N-terminal and C-terminal ends, which were maximally conserved with only 4 amino acid substitutions. In phylogenetic analysis no reasonable correlation between host species and/or geographic origin of the isolates was observed. Alignment with capsid protein genes of other Trichoviruses revealed the TaTao ACLSV peach isolate to be phylogenetically closest to Peach mosaic virus, Apricot pseudo chlorotic leaf spot virus and Cherry mottle leaf virus. Recombination analysis (RDP3 ver.2.6) done for all the available ACLSV complete CP sequences of the world and Indian isolates indicate no significant evidence of recombination. However, one recombination event among Indian ACLSV-CP isolates was detected. To the best of our knowledge, this is the first report of complete CP sequence variability study from India and also the first evidence of homologous recombination in ACLSV.

  9. 斑点杂交检测对虾白斑综合征病毒青岛株在螯虾体内的动态分布%Dynamic Distribution of Shrimp White spot syndrome virus (WSSV) Qingdao Strain in Crawfishes by Dot Hybridization

    朱建中; 夏晓勤; 陆承平; 郭福生

    2001-01-01

    A fragment sized 400bp of White spot syndrome virus(WS SV,formerly de signated NOSV),recovered from recombinant plasmid pAFD, was labeled with Digox igenin as a probe to detect dynamic distribution of WSSV within 120h and 72h in crawfishes(Cambarus proclarkii) inoculated WSSV by oral taking and injecti on r espectively. Stomach epithelium, intestine epithelium, heart, gill, haemolymph, muscle, hepatopancreas, hypoderm, connective tissue and ovary of infected crawfi shes were examined for WSSV. In both groups, WSSV was first detected in heamoly mph at 12h p.i. and then disappeared. Again it was detected at 96h p.i. only in oral infection group and maintained till 120h p.i., but it didn't appear at 72h p.i. in injection group. WSSV in heart, muscle was detected at 36h p.i. in oral infection group and 24h p.i. in injection group respectively, and then increased generally. In addition, WSSV in intestine epithelium, connective tissue, ovary of oral infection group and intestine epithelium, hypoderm, ovary of injection g roup could also be detected. In dead crawfishes after 120h and 72h p.i. in two groups, WSSV could be detected in all the examined tissues and it demonstrated t hat systemic infection occurred in the animales. The tissue containing more amo unts of WSSV was hypoderm in oral infection group, while intestine epithelium, g ill, hypoderm, ovary in injection infection group. It deduced that WSSV first a ppears in haemolymph and then goes into heart, muscle and other tissues and prol iferates in them. Once again, WSSV is released into heamolymph resulting in syst emic infection till crawfishes' death.

  10. 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

    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

  11. A prospective randomized clinical trial into the capacity of a toothpaste containing NovaMin to prevent white spot lesions and gingivitis during orthodontic treatment

    Hoffman, Derek A.; Clark, Andrew E.; Rody, Wellington J.; McGORRAY, SUSAN P.; Wheeler, Timothy T.

    2015-01-01

    Background White spot lesions and gingivitis represent common, yet challenging, dilemmas for orthodontists. Fluoride has shown some benefit as a protective measure against demineralization; however, this is usually insufficient for orthodontic patients with less than ideal oral hygiene. Dentifrices containing calcium sodium phosphosilicate bioactive glass (NovaMin) have been proposed to aid in prevention of white spot lesions and gingival inflammation. Thus, the purpose of this study was to d...

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

    Pamella Akoth Ogada

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

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

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

    2016-01-01

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

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

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

    2007-01-01

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

  15. The use of SD-OCT in the differential diagnosis of dots, spots and other white retinal lesions

    Zaharova E

    2011-10-01

    Full Text Available Elena Zaharova1, Jerome Sherman1-31State University of New York's State College of Optometry, University Eye Center, New York, NY, USA; 2SUNY Eye Institute, New York, NY, USA; 3New York Eye Institute and Laser Center, New York, NY, USAPurpose: To demonstrate the utility of a retinal imaging technique using spectral domain optical coherence tomography (SD-OCT for creating a B-scan layer-by-layer analysis to aid in the differential diagnosis of various retinal dots, spots, and other white lesions.Design: Review.Methods: A retrospective review of imaging studies performed with SD-OCT (Topcon, 3DOCT-2000, Oakland, NJ at SUNY State College of Optometry.Results: B-scan layer-by-layer analysis and unique SD-OCT reflectivity patterns of the following retinal white lesions are reviewed in the order of their retinal layer localization: myelinated nerve fiber layer, cotton wool spot, exudates, edema residues, drusen, fundus albipunctatus, Stargardt disease, Bietti crystalline dystrophy, punctate inner choroidopathy (PIC, presumed ocular histoplasmosis syndrome (POHS, post-photocoagulation chorioretinal scarring, and osseous choristoma.Conclusion: The reviewed images demonstrate the utility of SD-OCT in the identification of the unique characteristics of the presented retinal pathologies. SD-OCT is ideal for retinal layer localization of lesions, thus enhancing the differential diagnosis of retinal dots, spots, and other white lesions. Even though true pathognomonic patterns are rare, highly suggestive findings of certain retinal abnormalities often facilitate immediate recognition and diagnosis.Keywords: SD-OCT, photoreceptor integrity line, retinal pigment epithelium, white dot syndrome, retinal pathology, imaging

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

    Hae-Ryun Kwak

    2015-12-01

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

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

    Kwak, Hae-Ryun; Kim, Jeong-Soo; Cho, Jeom-Deog; Lee, Joong-Hwan; Kim, Tae-Sung; Kim, Mi-Kyeong; Choi, Hong-Soo

    2015-12-01

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

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

    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

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

    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.

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

    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.

  1. Deproteinization of tooth enamel surfaces to prevent white spot lesions and bracket bond failure: A revolution in orthodontic bonding

    Roberto Justus

    2016-01-01

    Full Text Available Orthodontic treatment success is jeopardized by the risk of development of white spot lesions (WSLs around orthodontic brackets. Unfortunately, the formation of WSLs still remains a common complication during treatment in patients with poor oral hygiene. Nearly 75% of orthodontic patients are reported to develop enamel decalcification because of prolonged plaque retention around brackets. It is the orthodontist′s responsibility to minimize the risk of patients having enamel decalcifications as a consequence of orthodontic treatment. This can be achieved by using hybrid, fluoride-releasing, glass ionomer cement to bond brackets, with deproteinization of the enamel surface before phosphoric acid etching.

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

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

    2015-04-01

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

  3. Immunity to nervous necrosis virus infections of orange-spotted grouper (Epinephelus coioides) by vaccination with virus-like particles.

    Lin, Kebing; Zhu, Zhihuang; Ge, Hui; Zheng, Leyun; Huang, Zhongchi; Wu, Shuiqing

    2016-09-01

    Nervous necrosis virus (NNV) is a kind of the betanodaviruses, which can cause viral nervous necrosis (VNN) and massive mortality in larval and juvenile stages of orange-spotted grouper (Epinephelus coioides). Due to the lack of viral genomes, virus-like particles (VLPs) are considered as one of the most promising candidates in vaccine study to control this disease. In this study, a type of VLPs, which was engineered on the basis of orange-spotted grouper nervous necrosis virus (OGNNV), was produced from prokaryotes. They possessed the similar structure and size to the native NNV. In addition, synthetic oligodeoxynucleotide (ODN) containing CpG motif was added in vaccines, and the expression patterns of several genes were analyzed after injecting with VLP and VLP with adjuvant (VA) to assess the regulation effect of vaccine for inducing immune responses. RT-PCR assays showed that six related genes in healthy tissues were ubiquitously expressed in all nine tested tissues. The vaccine alone was able to enhance the expression of genes, including MHCIa, MyD88, TLR3, TLR9 and TLR22 after vaccination, indicating that the vaccine was able to induce immune response in grouper. In liver, spleen and kidney, the gene expressions of VA group were all significantly higher than that of VLP group at 72 h post-stimulation, showing that the fish of VA challenge group obtained the longer-lasting protective immunity and resistance to pathogen challenge than that of VLP group. The data indicated that the efficacy of vaccine could be further enhanced by CpG ODN after vaccination and provided the reference for the development of future viral vaccine in grouper.

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

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

    2016-04-01

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

  5. Mitochondrial DNA population structure of the white-spotted Charr (Salvelinus leucomaenis) in the Lake Biwa water system.

    Kikko, Takeshi; Kuwahara, Masayuki; Iguchi, Kei'ichiro; Kurumi, Seiji; Yamamoto, Shoichiro; Kai, Yoshiaki; Nakayama, Kouji

    2008-02-01

    A phylogeographic analysis of mitochondrial DNA sequences was performed in order to elucidate the origin, dispersal process, and genetic structure of white-spotted charr in the Lake Biwa water system. Two haplotypes were most common in the Lake Biwa water system, and were also common in the adjacent inlet rivers of the Sea of Japan. These results suggest that in the glacial periods of the Pleistocene, white-spotted charr dispersed into the northern inlet rivers of Lake Biwa from adjacent inlet rivers of the Sea of Japan by watershed exchanges, colonizing the whole of the Lake Biwa water system. Mitochondrial DNA diversity contrasted sharply between the western and eastern parts of the system, suggesting that the populations in the western part might be more reduced than those in the eastern part in relation to the smaller habitat size. The high overall FST estimate (0.50), together with pairwise comparisons of FST, indicated significant genetic divergence between populations due to isolation and small population size. Hierarchical analysis (AMOVA) also showed that genetic variation was more pronounced among regions (28.39%) and among populations within regions (47.24%) than within populations (24.37%). This suggests that each population in and around the Lake Biwa water system should be treated as a significant unit for conservation and management.

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

    Hodi, Anna Maria

    2014-02-01

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

  7. Hypoxia increases susceptibility of Pacific white shrimp to whitespot syndrome virus (WSSV

    M. Lehmann

    2016-04-01

    Full Text Available The present study aimed to evaluate the mortality, reactive oxygen species production (ROS and total hemocyte counts (THC of the marine shrimp Litopenaeus vannamei infected with the white spot syndrome virus (WSSV at three levels of oxygen saturation. For this, 360 shrimp (20±2g were distributed in 24 tanks (60L, divided in two groups (infected and non-infected, which were subjected to 30, 60 and 100% of dissolved oxygen saturation (in quadruplicate. During 96 hours after infection, daily hemolymph samples were collected for hemato-immunological parameter evaluation (THC and ROS and dead animals were removed and computed to assess cumulative mortality rates. In the infected group, animals subjected to 100% saturation showed higher ROS production (P<0.05 after 48 hours, while THC was significantly reduced (P<0.05, regardless of oxygen saturation. The hypoxia resulted in high mortality when compared to 100% saturation condition. In the uninfected group, no significant differences were observed in all evaluated parameters. Thus, the hypoxia condition increased the susceptibility of shrimp to the infection of WSSV, which may be partly related to the low ROS production showed by the animals subjected to 30% oxygen saturation.

  8. Cause Analysis of White Spots Formed on Tinplate Surface%镀锡板表面小白点原因分析

    孙悦庆; 朱国和; 杨洁

    2012-01-01

    The forming cause of the white spots was preliminarily determined by SEM and EDS analyses as well as the observation of the micro-morphology and element types at the white spot defects. The analogue electroplating tests indicate that the oxides, oil stains and adhered rubber on strip steel surface are the cause for formation of the white spot defects.%通过对镀锡板表面产生的小白点进行扫描电镜和能谱分析,观察小白点缺陷处的微观形貌和存在的元素种类,初步确定小白点形成的原因.模拟电镀试验表明:带钢表面的氧化物、油污以及黏附的橡胶是产生小白点缺陷的原因.

  9. A comparison of the antigens present on the surface of virus released artificially from chick cells infected with vaccinia virus, and cowpox virus and its white pock mutant

    Baxby, Derrick

    1972-01-01

    Antisera prepared against vaccinia and cowpox viruses were absorbed with purified suspensions of vaccinia virus, red cowpox and white cowpox viruses. They were then tested for their ability to neutralize the viruses, and to precipitate the virus soluble antigens. The results showed that some virus specific antigens were not virus surface components and that some components were present on the surface of all three viruses. However, certain components were detected on the surface of vaccinia virus but not on the surface of cowpox virus, and vice versa. Some evidence for the existence of a vaccinia-specific surface component was also obtained. Comparisons between results of cross-neutralization tests and immunodiffusion tests on the absorbed sera indicated that antibody to a number of antigens, including the classical LS, and the cowpox-specific d antigen play no part in the process of poxvirus neutralization. ImagesFig. AFig. BFig. CFig. DFig. EFig. FFig. G PMID:4624399

  10. Spot phosphor concept applied to the remote phosphor configuration of a white phosphor-converted LED

    Acuña, Paula; Correia, António; Ryckaert, Jana; Meuret, Youri; Deconinck, Geert; Hanselaer, Peter

    2016-04-01

    Although the remote phosphor technology outperforms the conformal phosphor technology for mid-power applications, one of the limiting factors is the amount of phosphor required and its impact on the total cost. Besides, an important loss mechanisms in remote phosphor LED technology is the re-absorption of converted light. An obvious solution to this issue is enabling a light path for the converted light, such that further interactions with the phosphor element are avoided. In order to explore such a configuration, a simulation model of a phosphor element is devised and validated based on experimental data and the application of the inverse adding-doubling method. The resulting configuration, denoted as spot concept, along with a long-pass filter is shown to be a potential solution to reduce the phosphor usage. Since the moderate change in the light extraction ratio when applying the spot concept is partly attributed to the losses in the secondary optics needed to narrow the LED beam, the application of the spot concept configuration with a directional light source such as a laser diode could be a powerful combination for the enhancement of the light extraction ratio.

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

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

    2011-01-01

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

  12. Requirements for ER-Arrest and Sequential Exit to the Golgi of Tomato Spotted Wilt Virus Glycoproteins

    Ribeiro, D.M.O.G.; Goldbach, R.W.; Kormelink, R.J.M.

    2009-01-01

    The envelope glycoproteins Gn and Gc are major determinants in the assembly of Tomato spotted wilt virus (TSWV) particles at the Golgi complex. In this article, the ER-arrest of singly expressed Gc and the transport of both glycoproteins to the Golgi upon co-expression have been analyzed. While prel

  13. PRODUCTION OF POLYCLONAL ANTIBODIES AGAINST PELARGONIUM ZONATE SPOT VIRUS COAT PROTEIN EXPRESSED IN ESCHERICHIA COLI AND APPLICATION FOR IMMUNODIAGNOSIS

    Pelargonium zonate spot virus (PZSV), a new emerging disease on tomato in the United States, has been classified as the first member of new proposed genus, Anulavirus, within the family Bromoviridae and characterized as having unstable virions with weakly immunogenic properties. To develop serologic...

  14. 对虾白斑综合症的研究综述%Reserch on White Spot Syndrome of Penaeid Shrimp

    许雅香; 许梓荣

    2002-01-01

    对虾白斑综合症是由对虾白斑综合症病毒(white spot syndrome virus WSSV)引起的暴发性流行病。典型症状是病虾甲壳内侧出现0.5~2mm大小不等的白色斑点,此病病程短,死亡率高。白斑综合症病毒(WSSV)的形态结构与杆状病毒相似,杆状病毒分子分布于患病组织细胞核内。WSSV不仅水平传播,还存在垂直传播的可能性。到目前为上,诊断技术已较为进步,但尚无有效手段控制该病的发生。今后的研究重点在防治上。

  15. White spot lesions after orthodontic treatment assessed by clinical photographs and by quantitative light-induced fluorescence imaging; a retrospective study

    Beerens, M.W.; Boekitwetan, F.; van der Veen, M.H.; ten Cate, J.M.

    2015-01-01

    Objective. White spot lesions (WSL) are an important side-effect of orthodontic multi-bracket (MB) treatment. Standardized monitoring of such WSL may help in caries management. Materials and methods. In this retrospective study the discriminatory power of caries assessment on routine digital oral ph

  16. Effects of casein phosphopeptide amorphous calcium fluoride phosphate paste on white spot lesions and dental plaque after orthodontic treatment: a 3-month follow-up

    Beerens, M.W.; van der Veen, M.H.; van Beek, H.; ten Cate, J.M.

    2010-01-01

    The effects of casein phosphopeptide amorphous calcium fluoride phosphate (CPP-ACFP) paste vs. control paste on the remineralization of white spot caries lesions and on plaque composition were tested in a double-blind prospective randomized clinical trial. Fifty-four orthodontic patients, with multi

  17. The Vertical Structure of Major Meteorological Features on Jupiter: The Great Red Spot and White Ovals BC and DE

    Baines, Kevin H.; Carlson, Robert W.

    1999-01-01

    Multi-spectral imagery of Jupiter's Great Red Spot (GRS) and two White ovals acquired by the Galileo/NIMS are used to constrain the spatial variability of the vertical aerosol structure and the distribution of ammonia in and around these most-prominent anti-cyclonic features. All three features exhibit a high-altitude core spanning about 3/4 of their visual size when viewed with moderate absorption wavelengths, indicating a bulk elliptical, "wedding cake" shape in their overall three-dimensional cloud structure. A distinctive spiral pattern within the GRS core is seen in moderate methane and hydrogen absorption bandpasses. This pattern - which has been modelled to show a 2 km variation in cloudtop pressure within the GRS - is inconsistent with a different spiral-shaped pattern observed in ammonia-sensitive wavelengths, thus indicating spatial variability not only in the column abundance of ammonia within the GRS, but in its mixing ratio as well. White Ovals BC and DE were observed in February 1997, just a year before their unusual merger into a single feature. At the time of these observations, the centers of the two anti-cyclones were about 16 degrees apart, separated by a complex cyclonic feature which exhibited unusual spatial variability in its appearance in images acquired at ammonia-sensitive wavelengths. In particular, the northern half of this feature has the largest ammonia column abundance seen within the environs around the white ovals, indicating unusual variability in either cloud structure/altitude and/or ammonia humidity within the cyclone.

  18. Experimental susceptibility of Caspian white fish, Rutilus frisii kutum to Spring viraemia of carp virus.

    Zamani, H; Ghasemi, M; Hosseini, S M; Haghighi Karsidani, S

    2014-01-01

    Caspian white fish (Rutilus frisii kutum) is a fish of the family Cyprinidae, which is commercially harvested from the Caspian Sea. Experimental infection with Spring viraemia of carp virus (SVCV) was conducted in order to examine susceptibility of caspian White Fish and clinical impacts of infection. Fingerling fish were injected intra-peritoneally or waterborne-exposed with SVCV and were monitored daily for 7 weeks. Dead fish and those survived at the end of experimental period were collected for virus isolation and reverse transcriptase polymerase chain reaction analysis. Epithelioma papulosum cyprini cell line was used to re-isolate the virus and indirect fluorescent antibody test was conducted to identify the isolated virus. Infection trials showed that SVCV was highly pathogenic for the Caspian White Fish with mortality rate ranging from 75 to 85 %, depending on the viral challenge model. SVCV genome was detected from dead and apparently healthy fish tissues of both virus exposure models, which showed Caspian White Fish not only can be regarded as a susceptible host, but also serve as a vector of the virus.

  19. Haemolymph pathological research on white spot syndrome(WSS) of Litopenaeus vannamei%凡纳滨对虾白斑综合征血液病理研究

    冯守明; 杨先乐; 李军; 王蓉; 赵燕超

    2006-01-01

    对自然发病、投喂和注射感染的凡纳滨对虾白斑综合征(white spot syndrome, WSS)血液病理进行研究,结果发现:不同感染方式患病对虾的血液病理变化相似,表现为:1.患病对虾血细胞总数、透明细胞数量极显著减少,小颗粒细胞、大颗粒细胞极显著增加.2.显微病理变化主要表现为血涂片中血细胞明显减少且分布不均匀,破损或解体的细胞增多,呈典型的溶血状态.3.超微病理变化表现为,大部分血细胞坏死,少数血细胞呈不典型的凋亡.患病对虾的血细胞核中可见大量白斑综合征病毒(white spot syndrome virus, WSSV)粒子.病理变化表明血细胞是WSSV的主要靶细胞.

  20. Chlorotic spots on Clerodendrum, a disease caused by a nuclear type of Brevipalpus (Acari:Tenuipalpidae) transmitted virus

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

    2008-01-01

    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, Hi...

  1. Experimental susceptibility of Caspian white fish, Rutilus frisii kutum to Spring viraemia of carp virus

    Zamani, H.; Ghasemi, M.; Hosseini, S.M.; Haghighi Karsidani, S.

    2013-01-01

    Caspian white fish (Rutilus frisii kutum) is a fish of the family Cyprinidae, which is commercially harvested from the Caspian Sea. Experimental infection with Spring viraemia of carp virus (SVCV) was conducted in order to examine susceptibility of caspian White Fish and clinical impacts of infection. Fingerling fish were injected intra-peritoneally or waterborne-exposed with SVCV and were monitored daily for 7 weeks. Dead fish and those survived at the end of experimental period were collect...

  2. Protection of Penaeus monodon against white spot syndrome by continuous oral administration of a low concentration of Bacillus subtilis spores expressing the VP28 antigen.

    Pham, K-C; Tran, H T T; Van Doan, C; Le, P H; Van Nguyen, A T; Nguyen, H A; Hong, H A; Cutting, S M; Phan, T-N

    2017-03-01

    In this study, Bacillus subtilis spores expressing a chimeric protein, CotB-VP28, were used as a probiotic vaccine to protect black tiger shrimps (Penaeus monodon) against white spot syndrome virus (WSSV) infection. Oral administration of pellets coated with CotB-VP28 spores (at ≥1 × 10(9 ) CFU per g pellet) to shrimps induced immune-relating phenoloxydase activity (PO) in shrimps after 14 days of feeding (prior challenge) and at day 3 post challenge (1·26 and 1·70 fold increase respectively). A 75% protection rate was obtained by continuous feeding of the spore-coated pellets at ≥1 × 10(9 ) CFU per g for 14 days prior to WSSV challenge and during all the postchallenge period. Even when the amount of CotB-VP28 spores in feed pellets was reduced down to ≥5 × 10(7)  CFU per g and ≥1 × 10(6)  CFU per g, relatively high protection rates of 70 and 67·5%, respectively, were still obtained. By contrast, feeding pellets without spores (untreated group) and with naked spores (PY79 group) at ≥1 × 10(9)  CFU per g could not protect shrimps against WSSV. These data suggest that supplementation of CotB-VP28 spores at low dose of ≥1 × 10(6)  CFU per g could be effective as a prophylactic treatment of WSS for black tiger shrimps.

  3. Role of casein phosphopeptide amorphous calcium phosphate in remineralization of white spot lesions and inhibition of Streptococcus mutans?

    Vashisht, Ruchi; Indira, Rajamani; Ramachandran, S; Kumar, Anil; Srinivasan, Manali Ramakrishnan

    2013-01-01

    Introduction: To promote the remineralization by ionic exchange mechanism instead of invasive techniques many remineralizing agents can be used. Objective: To evaluate the remineralization effects of casein phosphopeptide-amorphous calcium phosphate (CPP-ACP) on white spot lesions (WSLs) and its inhibitory effect on Streptococcus mutans colonization. Materials and Methods: The study group consisted of 60 subjects exhibiting at least 1-WSL. Subjects were randomly divided into 2 groups: A test group using CPP-ACP cream (GC-Tooth Mousse, Leuven, Belgium) and a control group using only fluoride containing toothpaste for a period of 3-month. Baseline WSLs were scored using DIAGNOdent device (KaVo Germany) and the saliva samples were collected to measure S. mutans counts. After the 3-month period the WSLs were again recorded and the saliva collection was repeated. Result: DIAGNOdent measurements were increased by time (P = 0.002) in the control group and no statistically significant difference (P = 0.217) was found in the test group by the 3-month period. In both groups, the mutans counts were decreased in the 3-month experimental period. Conclusion: These clinical and laboratory results suggested that CPP-ACP containing cream had a slight remineralization effect on the WSL in the 3-month evaluation period however, longer observation is recommended to confirm whether the greater change in WSLs is maintained. PMID:23956538

  4. Role of casein phosphopeptide amorphous calcium phosphate in remineralization of white spot lesions and inhibition of Streptococcus mutans?

    Ruchi Vashisht

    2013-01-01

    Full Text Available Introduction: To promote the remineralization by ionic exchange mechanism instead of invasive techniques many remineralizing agents can be used. Objective: To evaluate the remineralization effects of casein phosphopeptide-amorphous calcium phosphate (CPP-ACP on white spot lesions (WSLs and its inhibitory effect on Streptococcus mutans colonization. Materials and Methods: The study group consisted of 60 subjects exhibiting at least 1-WSL. Subjects were randomly divided into 2 groups: A test group using CPP-ACP cream (GC-Tooth Mousse, Leuven, Belgium and a control group using only fluoride containing toothpaste for a period of 3-month. Baseline WSLs were scored using DIAGNOdent device (KaVo Germany and the saliva samples were collected to measure S. mutans counts. After the 3-month period the WSLs were again recorded and the saliva collection was repeated. Result: DIAGNOdent measurements were increased by time (P = 0.002 in the control group and no statistically significant difference (P = 0.217 was found in the test group by the 3-month period. In both groups, the mutans counts were decreased in the 3-month experimental period. Conclusion: These clinical and laboratory results suggested that CPP-ACP containing cream had a slight remineralization effect on the WSL in the 3-month evaluation period however, longer observation is recommended to confirm whether the greater change in WSLs is maintained.

  5. Observation of white spot lesions using swept source optical coherence tomography (SS-OCT): in vitro and in vivo study.

    Ibusuki, Takahide; Kitasako, Yuichi; Sadr, Alireza; Shimada, Yasushi; Sumi, Yasunori; Tagami, Junji

    2015-01-01

    This study aimed to assess swept source optical coherence tomography (SS-OCT) for in vitro and in vivo detection of enamel white spot lesion (WSL). WSLs without surface breakdown on 33 extracted human posterior teeth were non-invasively scanned using SSOCT. The teeth were then cross-sectioned and imaged under confocal laser scanning microscope (CLSM) and light microscopy (LM). SS-OCT cross-sectional images were compared with CLSM and LM. WSL shapes in SS-OCT images closely corresponded to those of LM. There were significant correlations (pSS-OCT and LM (r=0.92), SS-OCT and CLSM (r=0.80) and CLSM and LM (r=0.85). Six WSLs were also evaluated clinically using SS-OCT; clear in-depth images of these natural WSLs were obtained in vivo. SS-OCT appears to be an effective tool for observation of the internal structure of WSLs, enabling quantitative assessment of WSL depth. Such data can be considered in the clinical management of WSLs.

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

    Manuel Restrepo

    2015-01-01

    Full Text Available 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.

  7. Control of white spot lesion adjacent to orthodontic bracket with use of fluoride varnish or chlorhexidine gel.

    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.

  8. Fontes de resistência em tomateiro aos begomovírus bissegmentados Tomato yellow spot virus e Tomato severe rugose virus

    2012-01-01

    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 seve...

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

    Berniak Hanna

    2016-06-01

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

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

    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 μg g(-1) fish body weight, FBW) and immunizing frequencies (administration once, twice and thrice). A single vaccination with the dosage of 1.5 μg 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.

  11. Virus diseases risk-factors associated with shrimp farming practices in rice-shrimp and intensive culture systems in Mekong Delta Viet Nam

    Duc, P.M.; Tuyet Hoa, T.T.; Nguyen Thanh Phuong,; Bosma, R.H.; Huynh V., Hien; Tran N., Tuan

    2015-01-01

    In Mekong Delta, viral infection, including white spot syndrome virus (WSSV), monodon baculovirus (MBV), heptopancreatic parvovirus (HPV), infectious hypodermal and hematopoietic necrosis virus (IHHNV) and gill-associated nidovirus (GAV) frequently infect cultured shrimp starting at the postlarvae s

  12. Dried blood spot sampling for hepatitis B virus serology and molecular testing.

    Sofiane Mohamed

    Full Text Available BACKGROUND AIMS: Dried blood spots (DBS on filter paper have been successfully used to diagnose and monitor several infectious diseases. The aim was to investigate the performance of DBS in hepatitis B virus (HBV diagnosis using commercial tests in comparison to standard methods. METHODS: Paired DBS and plasma samples were collected from 200 patients: 100 patients with HBsAg negative status and 100 patients with HBsAg positive status. In the latter patient, HBeAg reactivity was tested. Ten samples of anti-HBs were collected from people vaccinated against HBV. We also studied 50 patients with positive HBV DNA viral load in plasma and 10 HBV DNA negative patients. HBV genotypes and gene polymerase mutations were determined in 10 randomly selected HBV-infected patients. The DBS sample consisted of 50 µL of whole blood, i.e. a 12-mm paper card. RESULTS: The sensitivity thresholds of HBsAg and anti-HBs antibody were 0.30 ± 0.08 IU/mL and 18.11 ± 6.05 IU/mL, respectively, for DBS with 98% sensitivity and 100% specificity. Sensitivity was 98% and specificity 100% for the detection of HBV DNA on a blotter, considering an HBV DNA threshold of 914.1 ± 157.8 IU/ml. Ten patients had an HBeAg positive status in plasma, all were detected positive using DBS. HBV genotyping and mutation detection were successfully performed on DBS, with full concordance between the 10 paired DBS and plasma samples. CONCLUSION: This study shows DBS is a reliable alternative to plasma specimens for quantifying and detecting HBsAg, anti-HBs, HBeAg and genotyping. DBS may increase the opportunities for HBV testing and treatment follow-up in hard-to-reach individuals.

  13. Effectiveness of MI Paste Plus and PreviDent fluoride varnish for treatment of white spot lesions: A randomized controlled trial

    Huang, Greg J.; Roloff-Chiang, Brie; Mills, Brian E.; Shalchi, Salma; Spiekerman, Charles; Korpak, Anna M.; Starrett, Jeri L.; Greenlee, Geoffrey M.; Drangsholt, Ross J.; Matunas, Jack C.

    2014-01-01

    Introduction White spot lesions are a common sequela of orthodontic therapy. In this parallel-group randomized trial, we assessed the effectiveness of 2 agents commonly used to ameliorate white spot lesions compared with a normal home-care regimen. Methods Patients aged 12 to 20 years were recruited from the offices of orthodontists and dentists who belonged to the Practice-based Research Collaborative in Evidence-based Dentistry network. The patients had their orthodontic appliances removed within the past 2 months and had at least 1 white spot lesion affecting their maxillary incisors. The subjects were randomized to 1 of 3 arms: (1) an 8-week regimen of MI Paste Plus (GC America, Alsip, Ill), (2) a single application of PreviDent fluoride varnish (Colgate Oral Pharmaceuticals, New York, NY), and (3) usual home care (control). Photographs were taken at enrollment and 8 weeks later. Two panels consisting of 5 dental professionals and 5 laypersons assessed the before-and-after pairs of photographs in a blinded fashion. Objective assessments and self-assessments were also performed. Results One hundred fifteen subjects completed the study; 34 were assigned to the MI Paste Plus group, 40 to the fluoride varnish group, and 41 to the control group. The mean improvements assessed by the professional panel were 21%, 29%, and 27% in the MI Paste Plus, fluoride varnish, and control groups, respectively. The results from the lay panel were 29%, 31%, and 25%, respectively. Objective improvements in the surface affected were 16%, 25%, and 17%, respectively; self-assessments of improvement were 37% in all 3 groups. No assessments indicated significant differences between subjects in the active arms compared with the control arm. Conclusions MI Paste Plus and PreviDent fluoride varnish do not appear to be more effective than normal home care for improving the appearance of white spot lesions over an 8-week period. PMID:23273358

  14. Paternity testing using the poisonous sting in captive white-spotted eagle rays Aetobatus narinari: a non-invasive tool for captive sustainability programmes.

    Janse, M; Kappe, A L; Van Kuijk, B L M

    2013-03-01

    A group of captive white-spotted eagle rays Aetobatus narinari produced 20 offspring, with an unknown father. Part of the poisonous sting was removed from each fish and DNA was extracted from the epidermis for paternity research using eight microsatellite markers of which four were from another species Aetobatus flagellum. This non-invasive sampling technique can be applied on all members of Myliobatiformes.

  15. BT-14钛合金舱口外盖白斑问题%White Spots of Some BT-14 Titanium Alloy Hatch

    周传忠; 俞川; 郭文龙; 范小龙; 于雷

    2013-01-01

    BT-14 titanium alloy plate is the structure material of some hatch product. By the inspection before welding, the titanium alloy surface presences some white spots. The white spots surface pattern, microstructure as well as mechanical property were analyzed. The influence of the white spots organization on titanium plate us⁃ability was studies with emphasis, and precaution in the production follow-up was put forword.%  BT-14钛板为某产品舱口盖结构用材料,焊前检查时,发现钛板表面有数量不等的白色圆状斑点。文章从白斑的外观形貌、微观组织以及力学性能等方面进行分析,重点研究了白斑组织对钛板使用性能的影响,并提出后续生产中的预防措施。

  16. Identification of defense-related genes associated with tomato Sw-7 line against Tomato spotted wilt virus in tomato through transcriptome analysis

    Tomato spotted wilt virus (TSWV) is a highly infectious tospovirus, and one of the most damaging plant viruses infecting tomatoes worldwide. Developing a tomato cultivar with TSWV-resistance would be the most effective approach for disease management. Comparative analysis of differential expression ...

  17. Postsynaptic alpha 2-adrenoceptors mediate melanosome aggregation in melanophores of the white-spotted rabbitfish (Siganus canaliculatus).

    Amiri, M H

    2009-01-01

    The present investigation was undertaken to study the nature of neuro-melanophore junction in the white-spotted rabbit fish Siganus canaliculatus. In vitro experiments using split fin preparation indicated that melanophores of S. canaliculatus 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 phentolamine (non-specific alpha adrenergic antagonist). The melanophore responses to repeated potassium stimulation (up to 20 stimuli) did not show any sign of fatigue. However, statistically significant enhancement was observed in responses to potassium that followed the first five stimulations. Adrenergic agonists acted in a time and concentration-dependent manner and their relative potency had the following rank order: clonidine (alpha-2 specific agonist) > norepinephrine (non-specific adrenergic agonist) > phenylephrine (alpha-1 specific agonist) > methoxamine (alpha-1-specific agonist). Yohimbine exerted a more potent inhibiting effect on norepinephrine induced melanosome aggregation compared to phentolamine. Prazosine (alpha-1 specific antagonist) had no effect on such aggregation. Chemically denervated melanophores displayed hypersensitivity to alpha-adrenergic agonists but were refractive to potassium ion stimulation. The refractivity of denervated melanophores to potassium indicates the effect of potassium ion is not direct on melanophores but it is rather through depolarization effect of potassium on the neuro-melanophore peripheral sympathetic fibers and hence release of norepinephrine. In denervated melanophores, similar to intact melanophores, only phentolamine and yohimbine but not prazosine, significantly inhibited melanosome aggregation effect of norepinephrine, indicating that norepinephrine effect is through postsynaptic alpha-2 adrenoceptors. The present data demonstrate

  18. Efficacy of Glass Ionomer Cements for Prevention of White Spot Lesions During Orthodontic Banding: A Randomized Clinical Trial

    Masoud Fallahinejad Ghajari

    2016-08-01

    Full Text Available Objectives: This study aimed to compare the incidence of white spot lesions (WSLs around orthodontic bands following the application of two glass ionomer (GI cements namely GC Gold Label and GC Fuji Plus for six to 12 months.Materials and Methods: A total of 186 permanent first molars of orthodontic patients requiring banding of at least two permanent first molars were chosen. The teeth were examined for caries and presence of WSLs by visual inspection and by DIAGNOdent (scores 0-29. Orthodontic bands were randomly cemented to the right or left molars using GC Gold Label or GC Fuji Plus GI cements. Samples were randomly divided into three groups and bands were removed after six, nine and 12 months in groups 1, 2 and 3, respectively. The teeth were then examined for caries and presence of WSLs by visual inspection. DIAGNOdent was used on the buccal and lingual surfaces to determine the presence of WSLs. The data were statistically analyzed using one-way ANOVA, multivariate repeated measures ANOVA, the Kruskal Wallis and the Mann-Whitney tests.Results: Totally 174 teeth were evaluated. DIAGNOdent scores were not significantly different before cementation and after removal of bands in buccal and lingual surfaces of the teeth in the two cement groups. Lesions simulating WSLs were seen in 21 out of 174 teeth but DIAGNOdent scores were not indicative of caries.Conclusion: Remarkable WSLs were not detected visually or by DIAGNOdent at six, nine or 12 months following the cementation of bands with two GI cements.

  19. Randomized controlled trial on fluoride varnish application for treatment of white spot lesion after fixed orthodontic treatment.

    Du, Minquan; Cheng, Ning; Tai, Baojun; Jiang, Han; Li, Jing; Bian, Zhuan

    2012-04-01

    The purpose of this study was to determine the efficacy of fluoride varnish (5% sodium fluoride, Duraphat(®), Colgate) in reverting white spot lesions (WSLs) after fixed orthodontic treatment. This study was a randomized, parallel group, controlled clinical trial. Using saline solution as control, 110 participants (mean age ± standard deviation: 16.6 ± 3.2 years) ranging from 12 to 22 years old were randomly assigned to either the test group (group 1) or the control group (group 2). Application of fluoride varnish or saline was applied onto tooth surfaces with WSLs every month during the first 6 months after debonding. The labial (buccal) surfaces of the teeth were assessed by the use of a DIAGNOdent pen (DD) at the baseline, 3- and 6-month follow-up visits. After 6 months, 96 subjects with a total of 209 study teeth (47 subjects, 104 teeth in group 1; 49 subjects, 105 teeth in group 2) remained. The WSLs had a mean DD reading at baseline of 17.66 ± 5.36 in group 1 and 16.19 ± 5.70 in group 2, which decreased by 5.78 and 2.44, respectively, at the 3-month follow-up visit and decreased by 7.56 and 3.09, respectively, at the 6-month follow-up visit. The mean baseline DD readings in the two groups were similar (t test, P > 0.05). There was statistically significant differences between the mean DD readings of the two groups at the 3-month (P orthodontic treatment.

  20. Effects of various remineralizing agents on the outcome of post-orthodontic white spot lesions (WSLs: a clinical trial

    Sombir Singh

    2016-08-01

    Full Text Available Abstract Background One of the most undesirable side effects of comprehensive orthodontic treatment is white spot lesions (WSLs. Despite many attempts at prevention of WSLs, its prevalence remains very high on debonding. There are many agents like fluoride toothpastes, fluoride varnishes, and fluoride mouth rinses, and casein phosphopeptide-amorphous calcium phosphate (CPP-ACP is frequently used for the remineralization of WSLs. However, there is no consensus in the literature with respect to the success rates of these agents. Thus, the present study was designed to evaluate the efficacy of fluoride toothpaste alone and in combination with fluoride varnish and CPP-ACP plus crème in the remineralization of post-orthodontic WSLs. Methods Forty-five subjects in the age range of 16–25 years having at least one post-orthodontic WSL were included in the study. All the subjects were randomly divided into three groups (toothpaste group, varnish group, and CPP-ACP group. The efficacy of various remineralizing agents on the remineralization of WSLs was evaluated clinically and by DIAGNOdent immediately after debonding and subsequently after 1, 3, and 6 months of their use. Results Twice daily use of fluoride toothpaste alone had no significant effect on remineralization of WSLs at various intervals of observations (P = 0.078. Application of fluoride varnish along with twice daily use of fluoride toothpaste for 6 months significantly decreased the severity of WSLs (P  0.05. Conclusions The use of fluoride varnish and CPP-ACP plus crème in addition to twice daily use of fluoride toothpaste had no additional benefit in the remineralization of post-orthodontic WSLs.

  1. Caspian White Fish (Rutilus frisii kutum) as a host for Spring Viraemia of Carp Virus.

    Ghasemi, M; Zamani, H; Hosseini, S M; Haghighi Karsidani, S; Bergmann, S M

    2014-06-04

    Rutilus frisii kutum is a fish of the Cyprinidae Family which is native in Caspian Sea and commercially cultured in Iran. This study was conducted to investigate susceptibility of Caspian White Fish to Spring Viraemia of Carp Virus (SVCV) infection and to evaluate influence of different challenge routes on virulence of the virus. Fingerlings were infected by immersion, intra-peritoneal (i.p.) injection, cohabitation and orally. Dead and surviving fish were collected for histological examination as well as for virus re-isolation by cell culture, Reverse Transcriptase Polymerization Chain Reaction (RT-PCR) and Indirect Fluorescent Antibody Test (IFAT) analysis. The results indicated that immersion was the best infectious route of transmission with the highest mortality, whereas oral transmission showed the lowest mortality. The virus was also re-isolated from dead fish and identified by IFAT. In addition, histopathological changes including branchial, hepatic and splenic necrosis as well as glomerulonephritis and necrosis in kidney were observed in diseased fish tissues but not in the survivors. RT-PCR on samples obtained from surviving fish tissues detected viral genome in the fish surviving from immersion, i.p. injection and cohabitation challenges but not in the fish infected orally. In conclusion, Caspian White Fish are susceptible to infection by SVCV and virulence of the virus could be influenced by route of transmission. In addition, SVCV could persist in surviving fish, which may serve as reservoirs of the virus, transmitting infection to healthy fish population.

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

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

    2016-01-01

    Thrips-transmitted tospoviruses are economically important viruses affecting a wide range of field and horticultural crops worldwide. Tomato spotted wilt virus is the type member of the Tospovirus genus with a broad host range of more than 900 plant species. Interactions between these viruses and their plant hosts and insect vectors via RNA interference pathways are likely a key determinant of pathogenicity. The current investigation, for the first time, compares biogenesis of small RNAs be...

  3. Pupil - white spots

    ... disease can be fatal. When to Contact a Medical Professional Contact your health care provider if you notice any color changes in the ... to Expect at Your Office Visit The health ... exam and ask about your symptoms and medical history. The physical exam will include a detailed ...

  4. 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

    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.

  5. 凤仙花坏死斑病毒及其防治%Impatiens necrotic spot virus and its control

    汪鑫; 郭京泽

    2004-01-01

    凤仙花坏死斑病毒(Impatiens necrotic spot virus)简称INSV,是危害观赏植物的重要病害之一,它最早从有症状的凤仙花上分离鉴定出来,能侵染许多植物。凤仙花坏死斑病毒能随着带毒植物种苗广泛传播,经研究证明昆虫是这种病毒的重要传毒介体。这

  6. Monthly occurrence and prevalence of viral haemorrhagic septicaemia virus (VHSV) in whiting Merlangius merlangus.

    Altuntaş, C; Ogut, H

    2010-01-25

    A field survey was carried out to determine the occurrence and spread of viral haemorrhagic septicaemia virus (VHSV) in whiting Merlangius merlangus (L.), a key prey species in the southeastern Black Sea region. In March, April, September and December 2007 and monthly in 2008, 70 to 105 M. merlangus captured by gill nets were examined in pools of 7 fish per pool (total number of fish = 1603). After determining the seasonal occurrence of VHSV in whiting, 43, 80 and 88 whiting were sampled individually in January, February and March 2009, respectively, to determine age and size of whiting commonly infected with VHSV. The samples were tested in duplicate using BF-2 and EPC cell lines for virus isolation. Positive cultures (those demonstrating cytopathic effect, [CPE]), were tested further with a commercial ELISA kit to confirm that the observed CPE was caused by VHSV. VHSV was detected in M. merlangus only between February and April following a drop in the surface water temperature to 9 degrees C. All infected fish from the sampled whiting were asymptomatic carriers with low levels of virus. The estimated prevalence from pooled samples ranged between 1.35% (2.5 to 97.5 percentiles; 0.04 to 7.32%) to 3.14% (0.36 to 10.95%), whereas prevalence estimated from individual sampling was 7, 5 and 1% in January, February and March, respectively. Individual sampling also revealed that prevalence was higher among young fish (1 to 1.5 yr old) and decreased with increasing age. This is the first report of VHSV infection in whiting in the Black Sea.

  7. Dietary chitosan nanoparticles protect crayfish Procambarus clarkii against white spot syndrome virus (WSSV) infection.

    Sun, Baozhen; Quan, Haizhi; Zhu, Fei

    2016-07-01

    Chitosan nanoparticles have exhibited potential antibacterial activity or anticancer activity as their unique character. In this study, we investigated the effect of chitosan nanoparticles protect crayfish Procambarus clarkii against WSSV. Chitosan (from crab shell) nanoparticles were prepared by ultrafine milling. The physicochemical properties of the nanoparticles were determined by particle size measure, zeta potential analysis and scanning electron microscope observation. The total hemocyte count (THC), phenoloxidase (PO) and superoxide dismutase (SOD) activity were measured at days 1, 4, 9 and 12, and the survival rate was also recorded after WSSV challenge. The results showed that chitosan nanoparticles could enhance the survival rate of WSSV-challenged crayfish. And crayfish fed diets supplemented with 10 mg/g chitosan nanoparticles (65% mortality) showed a significantly higher survival rate when compared to the control group (100% mortality). The analysis of immunological parameters revealed that 10 mg/g chitosan nanoparticles showed significantly higher level of prophenoloxidase (proPO), superoxide dismutase (SOD) and total hemocyte count (THC) when compared to the control group. It was found that chitosan nanoparticles could inhibit WSSV replication in crayfish. Our results demonstrated that dietary chitosan nanoparticles effectively improve innate immunity and survival of P. clarkii challenged with WSSV.

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

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

    concentrations were found to be effective in controlling and treating the viral infection. However, 15 ppm concentration was found to be more effective as improvements in both clinical and behavioural aspects were noticed within two days of the treatment...

  9. PENYEBARAN DAN PREVALENSI WHITE SPOT SYNDROME VIRUS (WSSV PADA BUDI DAYA UDANG WINDU (Penaeus monodon

    Muliani Muliani

    2016-11-01

    The aims of this experiment was to know the distribution and prevalences of WSSV on tiger shrimp. Sample collection and WSSV detection conducted with PCR method was carried out during April 2004 to November 2006. Tiger shrimp broodstock samples were collected from Central Java, South Sulawesi, Gorontalo, Kalimantan, and Timika waters, and the other samples (tiger shrimp post larvae, juveniles, and cultured shrimp were collected from several region in South Sulawesi. The pleopod, pereiopod, eye stalk, carapax, gill, tail, muscle of broodstock, juveniles, and cultured shrimp, and 30 pcs of postlarvae were extracted using lysis buffer to collect genomic DNA. WSSV DNA amplification was carried out using first and nested PCR technique by specific sequence amplification kit (IQ2000TM  Detection and Prevention system. The WSSV DNA was visualized by gell documentation system. The result showed that more than 33% of broodstock resources of Indonesia waters and 90% of shrimp culture area of South Sulawesi were contaminated by WSSV. WSSV was also infected tiger shrimp broodstock, postlarvae, juveniles, and tiger shrimp cultured with the highest prevalence (40.4% was on tiger shrimp cultured and the lowest prevalence (5.4% was on postlarvae.

  10. PENYEBARAN DAN PREVALENSI WHITE SPOT SYNDROME VIRUS (WSSV) PADA BUDI DAYA UDANG WINDU (Penaeus monodon)

    2016-01-01

    Penelitian ini bertujuan untuk mengetahui penyebaran dan tingkat prevalensi serangan WSSV pada budi daya udang windu. Pengumpulan sampel dan deteksi WSSV dengan teknik PCR dilakukan dari bulan April 2004 sampai November 2006. Sampel induk udang windu yang dikumpulkan berasal dari perairan Jawa Tengah, Sulawesi Selatan, Gorontalo, Kalimantan Timur, dan Timika. Sedangkan benur, tokolan, dan udang yang dibudidayakan dikumpulkan dari beberapa lokasi di Sulawesi Selatan. Sampel udang diambil bagia...

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

    Tu, Jiagang; Chen, Wenjie; Fu, Xiaozhe; Lin, Qiang; Chang, Ouqin; Zhao, Lijuan; Lan, Jiangfeng; Li, Ningqiu; Lin, Li

    2016-05-19

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

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

    Jiagang Tu

    2016-05-01

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

  13. Research advances of Melon necrotic spot virus%甜瓜坏死斑点病毒研究进展

    吴会杰; 古勤生

    2015-01-01

    Cucumis melo is one of the important Cucurbitaceae in the tropical and temperate regions and also a major eco⁃nomic crop in our country. Melon necrotic spot virus(MNSV)cause significant economic losses in melon. The taxonomic sta⁃tus,characteristics and distribution,analysis of genes and pathogenicity,virus cell-to-cell movement and influence factor of transmission and resistance of MNSV were reviewed. Although there were many researches on MNSV,but there are few re⁃ports on the virus pathogenic mechanism. Therefore,it is necessary to find the host factors related to the virus and reveal the pathogenicity mechanisms of MNSV.%甜瓜(Cucumis melo)是世界上热带和温带地区的一种重要葫芦科作物,也是我国重要的经济作物,甜瓜感染甜瓜坏死斑点病毒(Melon necrotic spot virus,MNSV)会造成重大经济损失。本文对甜瓜坏死斑点病毒的分类地位、特性及发生分布、基因与致病性研究、病毒在细胞间的运动、影响传播因素及抗性研究等进展做一综述。目前,关于该病毒的研究国内外已有不少报道,但病毒致病机制的研究仍在探索,因此揭示其致病机制成为研究该病毒的首要任务,为该病毒的抗性研究奠定基础。

  14. Study on the Distribution of Disease-Resistant Shrimp Identified by DNA Markers in Respect to WSSV Infection in Different Seasons Along the Entire East Coast of India Aiming to Prevent White Spot Disease in Penaeus monodon.

    Mallik, A; Chakrabarty, U; Dutta, S; Mondal, D; Mandal, N

    2016-02-01

    White spot disease caused by white spot syndrome virus (WSSV) is responsible for harming shrimp aquaculture industry and results in a pandemic throughout the world. Undeniably, the knowledge on geographic distribution, transmission, virulence, and seasonal prevalence of this disease alongside information on the distribution of disease-resistant shrimps may be helpful to understand important aspects of disease biology. This study was intended to estimate WSSV prevalence by qualitative and quantitative PCR method among the Penaeus monodon samples collected from four different places namely Digha, West Bengal; Chilika, Orissa; Visakhapatnam, Andhra Pradesh; and Chennai, Tamil Nadu at three different seasons in the period of 2011-2013 from east coast of India. Along with this, the disease-resistant prevalence was also investigated using earlier developed 71 bp microsatellite and 457 bp RAPD-SCAR DNA marker among the collected shrimps. Qualitative PCR depicted that the cumulative WSSV prevalence at four places was the lowest (0%) at pre-monsoon, whereas, it was the highest (21.2%) during post-monsoon season. Quantitative real-time PCR showed the average copy number of WSSV to be the highest (~10(3) copy μg(-1) shrimp genomic DNA) at post-monsoon season. Additionally, estimated disease-resistant prevalence was the highest in Visakhapatnam (79%) and lowest in Digha (21%). It is well known to all that a trait cannot be identified using a single genetic pattern. This study will significantly contribute insight to develop specific pathogen-resistant (SPR) seeds of P. monodon simultaneously using two DNA markers that would be a cost-effective and safer approach towards disease prevention instead of conventional trends of seed generation from unselected wild broodstock.

  15. White matter tract injury and cognitive impairment in human immunodeficiency virus-infected individuals.

    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

    2009-04-01

    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 neuropsychological evaluation to investigate the relationship of white matter integrity to cognitive impairment and antiretroviral treatment variables. Participants included 39 HIV-infected individuals (49% with acquired immunodeficiency syndrome [AIDS]; mean CD4 = 529) and 25 seronegative subjects. Diffusion tensor imaging indices were mapped onto a common whole-brain white matter tract skeleton, allowing between-subject voxelwise comparisons. The total HIV-infected group exhibited abnormal white matter in the internal capsule, inferior longitudinal fasciculus, and optic radiation; whereas those with AIDS exhibited more widespread damage, including in the internal capsule and the corpus callosum. Cognitive impairment in the HIV-infected group was related to white matter injury in the internal capsule, corpus callosum, and superior longitudinal fasciculus. White matter injury was not found to be associated with HIV viral load or estimated CNS penetration of antiretrovirals. Diffusion tensor imaging was useful in identifying changes in white matter tracts associated with more advanced HIV infection. Relationships between diffusion alterations in specific white matter tracts and cognitive impairment support the potential utility of diffusion tensor imaging in examining the anatomical underpinnings of HIV-related cognitive impairment. The study also confirms that CNS injury is evident in persons infected with HIV despite effective antiretroviral treatment.

  16. Remineralization effect of casein phosphopeptide-amorphous calcium phosphate on enamel white spot lesions. A quantitative energy dispersive X ray elemental analysis: An in vitro study

    Fabrizio Guerra

    2014-06-01

    Full Text Available Background: The objective of this study was to evaluate, by means of elemental analysis the mineral density, calcium, and phosphorus weight percent of sound enamel, demineralized and CPP-ACP treated enamel. Elemental analysis allows elemental and isotopic composition of a biologic sample. It can be qualitative (determining what elements are present, and quantitative (determining how much of each are present. INCA Energy 250, Oxford Analytical Instruments Ltd. (UK, energy-dispersive X-ray spectroscopy system for elemental analysis was performed on random assigned samples. Methods: 12 sound premolars were extracted for orthodontic reason. Each tooth was sectioned by using a double-faced diamond microtome under water cooling into three section for a total of 36 samples and randomly assigned to three groups: Group 1 (control, Group 2 (WS: white spot , Group 3 (WST white spot treated of 12 samples each. Samples (Group 2 and Group 3 underwent equally to 24 h and 48 h of acid bath duration. Then all the treated samples (Group 3 were coated with CPP-ACP for 5 min before immersion into water twice a day. Group 2 served as control for enamel damage evaluation. Inca Point & ID, an analytic platform software for SEM was used for elemental analysis on samples from Group 1 (C, 2 (WS and Group 3 (WST in order to determine the weight % and atomic % presence of Ca and P. Results: The results of the samples analysis from the three Groups show different weight % and atomic% of Ca and P, and clearly reflect the different mineralization rates. Conclusions: 10% Casein phosphopeptide-amorphous calcium phosphate (CPP-ACP complex, promotes remineralization in vitro. The results of this in vitro study completely agree with this statement. Clinical studies to investigate the intraoral effectiveness of topical applications of CPP-ACP on white spot lesions are required to confirm these results.

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

    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.

  18. 十二指肠白点综合征的内镜诊断%Endoscopic diagnosis of duodenal white spot syndrome

    李保会; 田京灵; 余睿; 王志雄; 祝小林; 董晓菁; 李晓荣

    2014-01-01

    Objective To discuss the standard of endoscopic diagnosis of duodenal white spot syndrome and to test its accuracy by pathological diagnosis standard. Methods A total of 6,995 patients undergoing gastroscopy were detailedly examined through gastrofiberscope or electrogastroscope and endoscopic diagnosis were made by the standard of endoscopic diagnosis of duodenal white spot syndrome. Then, pathological examination was done to test and verify the accuracy of the standard of endoscopic diagnosis. Results There were 533 patients suffering from duodenal white spot pathological changes, accounting for 7.62%. And duodenal mucositis was confirmed by pathological findings in all 90 cases of the patients undergone their biopsy at the same time. Conclusions The appropriate standard of endoscopic diagnosis can improve the detection rate of duodenal white spot syndrome and there is a high coincidence rate between endoscopic diagnosis and pathological diagnosis.%目的:探讨十二指肠白点综合征的内镜诊断标准,并以病理诊断确定所用标准的准确率,以便确定并推广合适的内镜诊断标准,从而提高十二指肠白点综合征诊断率和准确率。方法:对6995例行纤维胃镜或电子胃镜检查的患者,根据所定的十二指肠白点综合征的内镜诊断标准进行观察诊断,并对同意病理活检者活体组织进行病理检查,比较内镜诊断与病理诊断的符合率。结果:在6995例行胃镜检查的患者中533例内镜诊断为十二指肠白点综合征,占7.62%;其中90例患者行病理活检,结果均证实为十二指肠黏膜炎症性病变。结论使用合适的内镜诊断标准可以使十二指肠白点综合征的内镜诊断具有较高的检出率且有很高的病理诊断符合率。

  19. Characterization of H3N6 avian influenza virus isolated from a wild white pelican in Zambia.

    Simulundu, Edgar; Mweene, Aaron S; Tomabechi, Daisuke; Hang'ombe, Bernard M; Ishii, Akihiro; Suzuki, Yuka; Nakamura, Ichiro; Sawa, Hirofumi; Sugimoto, Chihiro; Ito, Kimihito; Kida, Hiroshi; Saiwana, Lewis; Takada, Ayato

    2009-01-01

    We characterized an influenza virus isolated from a great white pelican in Zambia. Phylogenetic analysis showed that all of its gene segments belonged to the Eurasian lineage and that they appear to have evolved in distinct geographical regions in Europe, Asia, and Africa, suggesting reassortment of virus genes maintained in wild aquatic birds whose flyways overlap across these continents. It is notable that this virus might possess some genes of the same origin as those of highly pathogenic H7 and H5 viruses isolated in Eurasia. The present study underscores the need for continued monitoring of avian influenza viruses in Eurasia and Africa.

  20. Egg whites from eggs of chickens infected experimentally with avian hepatitis E virus contain infectious virus, but evidence of complete vertical transmission is lacking.

    Guo, H; Zhou, E M; Sun, Z F; Meng, X-J

    2007-05-01

    Avian hepatitis E virus (HEV) is genetically and antigenically related to human HEV. Vertical transmission of HEV has been reported in humans, but not in other animals. In this study, we showed that avian HEV could be detected in chicken egg-white samples. Subsequently, avian HEV in egg white was found to be infectious, as evidenced by the appearance of viraemia, faecal virus shedding and seroconversion in chickens inoculated with avian HEV-positive egg white, but not in chickens inoculated with HEV-negative egg white. To further assess the possibility of vertical transmission of avian HEV, batches of embryonated eggs from infected hens were hatched, and hatched chicks were monitored for evidence of avian HEV infection. However, no virus was detected in samples collected from the hatched chicks throughout this study, suggesting that avian HEV could not complete the vertical transmission cycle. The possible implications of our findings are also discussed.

  1. 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

    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.

  2. Effects of casein phosphopeptide amorphous calcium fluoride phosphate paste on white spot lesions and dental plaque after orthodontic treatment: a 3-month follow-up.

    Beerens, M W; van der Veen, M H; van Beek, H; ten Cate, J M

    2010-12-01

    The effects of casein phosphopeptide amorphous calcium fluoride phosphate (CPP-ACFP) paste vs. control paste on the remineralization of white spot caries lesions and on plaque composition were tested in a double-blind prospective randomized clinical trial. Fifty-four orthodontic patients, with multiple white spot lesions observed upon the removal of fixed appliances, were followed up for 3 months. Subjects were included and randomly assigned to either CPP-ACFP paste or control paste, for use supplementary to their normal oral hygiene. Caries regression was assessed on quantitative light-induced fluorescence (QLF) images captured directly after debonding and 6 and 12 wk thereafter. The total counts and proportions of aciduric bacteria, Streptococcus mutans, and Lactobacillus spp. were measured in plaque samples obtained just before debonding, and 6 and 12 wk afterwards. A significant decrease in fluorescence loss was found with respect to baseline for both groups and no difference was found between groups. The size of the lesion area did not change significantly over time or between the groups. The percentages of aciduric bacteria and of S. mutans decreased from 47.4 to 38.1% and from 9.6 to 6.6%, respectively. No differences were found between groups. We observed no clinical advantage for use of the CPP-ACFP paste supplementary to normal oral hygiene over the time span of 12 wk.

  3. Response of white leghorn chickens to infection with avian leukosis virus subgroup J and infectious bursal disease virus.

    Williams, Susan M; Sellers, Holly S

    2012-03-01

    The effects of viral-induced immunosuppression on the infectious status (viremia and antibody) and shedding of avian leukosis virus (ALV) were studied. Experimental white leghorn chickens were inoculated with ALV subgroup J (ALV-J) and infectious bursal disease virus (IBDV) at day of hatch with the ALV-J ADOL prototype strain Hcl, the Lukert strain of IBDV, or both. Appropriate groups were exposed a second time with the Lukert strain at 2 wk of age. Serum samples were collected at 2 and 4 wk of age for IBDV antibody detection. Samples for ALV-J viremia, antibody detection, and cloacal shedding were collected at 4, 10, 18, and 30 wk of age. The experiment was terminated at 30 wk of age, and birds were necropsied and examined grossly for tumor development. Neoplasias detected included hemangiomas, bile duct carcinoma, and anaplastic sarcoma of the nerve. Control birds and IBDV-infected birds were negative for ALV-J-induced viremia, antibodies, and cloacal shedding throughout experiment. By 10 wk, ALV-J-infected groups began to develop antibodies to ALV-J. However, at 18 wk the incidence of virus isolation increased in both groups, with a simultaneous decrease in antibody levels. At 30 wk, 97% of birds in the ALV-J group were virus positive and 41% were antibody positive. In the ALV-J/IDBV group, 96% of the birds were virus positive at 30 wk, and 27% had antibodies to ALV-J. In this study, infection with a mild classic strain of IBDV did not influence ALV-J infection or antibody production.

  4. Research on Causes and Precautions of White Spots and White Lines on Hard Anodized Surface%硬质阳极化表面白点白道原因及预防措施

    马永红

    2015-01-01

    通过对铝制精密活门偶件的加工工艺方法的研究,分析了目前铝制精密活门偶件硬质阳极化表面产生白点白道缺陷的原因及预防措施,解决长期以来铝制精密活门偶件一次交检合格率低的问题,提升了精密偶件类的加工能力。%The study reported the causes and precautions of white spots and white lines defect on hard anodized surface of aluminum precision valve coupled parts by research on processing method. The problem of low rate of qualified inspection for aluminum precision valve coupled parts was resolved. The processing capacity for precision parts was improved.

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

    Punya Nachappa

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

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

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

    2000-01-01

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

  7. First report of a resistance-breaking strain of Tomato spotted wilt virus infecting tomatoes with the Sw-5 tospovirus-resistance gene in California

    Management of Tomato spotted wilt virus (TSWV) with the Sw-5 resistance gene in tomato is highly effective. However, in certain regions of the world where resistant tomatoes have been continually planted, resistance-breaking strains of TSWV have emerged. In spring 2016 resistant tomatoes were obse...

  8. Characterization of two different apricot latent virus variants associated with peach asteroid spot and peach sooty ringspot diseases.

    Gentit, P; Foissac, X; Svanella-Dumas, L; Peypelut, M; Candresse, T

    2001-08-01

    Peach asteroid spot (PAS) and peach sooty ringspot (PSRS) are two diseases of stone fruit trees of unknown aetiology. The use of a cRNA probe of the newly described Apricot latent virus (ApLV), a tentative member of the Foveavirus genus, indicated the presence of cross-hybridizing agents in PAS isolate LA2 and in PSRS isolates Caserta 12 and Clava J4. Analysis of dsRNA patterns revealed in each case the presence of a major dsRNA band of about 9.6 kbp. The purified dsRNAs were used to obtain cDNA clones for isolates LA2 and Caserta 12. Sequence analysis of a 1.1 kbp cDNA clone from isolate LA2 showed very high homology with the known ApLV sequence, indicating that this isolate represents a closely related variant of ApLV. Sequence analysis of a 3.06 kbp Caserta 12 cDNA clone representing the 3' region of the genome revealed a genomic organization similar to that reported for other members of the Foveavirus genus, including the triple gene block and a large, 43.6 kDa coat protein. Sequence comparison with the CP gene of ApLV, the only sequenced region so far for this virus, showed an overall homology of 78%. These results indicate that the foveavirus represented by the Caserta 12 isolate of PSRS disease may be regarded as a distant variant of ApLV. The present results indicate that the viral agents associated with peach asteroid spot and peach sooty ringspot diseases might be variants of the recently described ApLV.

  9. BONE STRENGTH (ossis tibiae OF NATIVE PIGS ZŁOTNICKA SPOTTED BREED AND CROSSBREDS OF POLISH LARGE WHITE AND PoLISH LANDRACE PIGS

    Wojciech KAPELAŃSKI

    2012-01-01

    Full Text Available The purpose of this study was to compare the bone strength traits in two groups of pigs different in terms of genetic value, fast growth capability and meat deposition in the body. The study covered 33 fatteners of the Złotnicka Spotted breed (ZŁP and 20 crossbreds F1 (Polish Large White x Polish Landrace. Tibial bones were obtained following slaughter and evaluated in terms of properties and geometry, as well as bending and compressive strength. Mineral composition of the bone tissue was also established, i.e.: ash, Ca, P, Na, K, Zn and Mg. The significance of differences between the traits demonstrated by pigs in both groups subject to the study was estimated, with calculated overall correlations between the primary bone properties. The results confirmed significant differences in terms of geometry of the bones in both animal groups. In the ZŁP breed pigs, compared to the F1 crossbred group (PLW x PL, the outside and inside diameters of the tibial shaft were smaller (P ≤ 0.01, yet the average thickness of the wall of the bone was slightly larger. The bending force required to fracture the bone turned out to be lower in Złotnicka Spotted pigs, the opposite of the results obtained with regard to the compressive strength. Furthermore, calcium content, calcium-phosphorus ratio (Ca : P and sodium content were highly statistically or significantly larger in the bone tissue of the Złotnicka Spotted pigs. Consequently, the final results show that the bone strength demonstrated by modern pig breeds subject to intensive selection focused on high growth rate and increased meat deposition is in no way lower than the same trait recognized in the Złotnicka Spotted breed kept in preservative breeding conditions.

  10. Transient asymptomatic white matter lesions following Epstein-Barr virus encephalitis

    Yoo Young Jang

    2011-09-01

    Full Text Available We present the case of a patient with Epstein-Barr virus (EBV encephalitis who developed abnormal white matter lesions during the chronic phases of the infection. A 2-year-old-boy was admitted for a 2 day history of decreased activity with ataxic gait. The results of the physical examination were unremarkable except for generalized lethargy and enlarged tonsils with exudates. Brain magnetic resonance imaging (MRI at admission showed multiple high signal intensities in both basal ganglia and thalami. The result of EBV polymerase chain reaction (PCR of the cerebral spinal fluid was positive, and a serological test showed acute EBV infection. The patient was diagnosed with EBV encephalitis and recovered fully without any residual neurologic complications. Subsequently, follow-up MRI at 5 weeks revealed extensive periventricular white matter lesions. Since the patient remained clinically stable and asymptomatic during the follow-up period, no additional studies were performed and no additional treatments were provided. At the 1-year follow-up, cranial MRI showed complete disappearance of the abnormal high signal intensities previously seen in the white matter. The patient continued to remain healthy with no focal neurologic deficits on examination. This is the first case of asymptomatic self-limited white matter lesions seen in serial MRI studies in a Korean boy with EBV encephalitis.

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

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

    2014-01-01

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

  12. 铝合金微弧氧化膜白色斑点成因分析%Analysis of white spot formation on the micro-arc oxidation coating of aluminum alloy

    王青; 裴舒; 白利红; 武海玲; 苗成

    2011-01-01

    When micro-arc oxidation coatings were formed on aluminum alloy in silicate system electrolyte, white spots often appear on the coating, and the quality and appearance of work piece will be impaired. The composition of the white spots was analyzed by EDS and XRD. It is found that the white spots is SiO2. Furthermore, the formation reason was analyzed.%用硅酸盐体系电解液进行铝合金微弧氧化时,氧化膜表面出现白色斑点状物,影响零件质量和外观,对这种白色斑点状物进行EDS和XRD成分测试,同时还对形成原因进行分析.结果表明,白色斑点状物是由SiO2聚集而成.

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

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

    2016-01-01

    centers, a prison and outpatient clinics and included blood donors as negative controls. Five drops of finger capillary blood were spotted on filter paper, and a venous blood sample was obtained. The samples were analyzed for HBsAg, anti-HBc, anti-HBs, anti-HCV, and anti-HIV levels as well as subjected......, but correctly classified 95% of the anti-HCV-positive patients with chronic and past infections. Anti-HBc and anti-HBS showed low sensitivity in DBS (68% and 42%). CONCLUSION: DBS sampling, combined with an automated analysis system, is a feasible screening method to diagnose chronic viral hepatitis and HIV...

  14. Protective immunity of orange-spotted grouper(Epinephelus coioids) against a nervous necrosis virus isolated from China, and determination of the complete sequences of the virus

    CHEN Xiaoyan; WENG Shaoping; CHEN Shengping; CHEN Zhaoming; HE Jianguo

    2007-01-01

    On the basis of the sequence and analysis of genome from the orange-spotted nervous necrosis virus(OGNNV), China strain, a pair of special primers were designed according to the nucleotide sequences of RNA2 from OGNNV. The major capsid protein(MCP)gene of OGNNV was cloned by means of reverse-transcriptase polymerase chain reaction (RT-PCR)and ligated into the pET32a expression plasmid. The MCP gene of OGNNV was 1 017 bases, encoded a protein of 338 amino acid with a molecular mass of 37.1 kDa. Recombinant protein with a molecular mass of 57.4 kDa was expressed in E.coli BL21(DE3). Vaccine was prepared from the recombinant protein expressed in recombinant cells. The juvenile orange-spotted groupers (8 cm in average length) were immunized by intraperitoneal injection. Group A was challenged with infected tissue filtrates 25 d post-vaccination. The mortality in the vaccined group (A1, 30%) was a little higher than the unvaccined group (B2, 27.8%). Group B was challenged after three vaccine injections. The mortality in the vaccined group (B1, 16.7%) was lower than the unvaccined group (B2, 27.8%), And the relative percentage survival (RPS) value of vaccined group, compared with the unvaccined group, was 40%. The anti-recombinant protein sera with a 1∶100 dilution were mixed with double volume of infected tissue filtrates and incubated at 4 ℃ for 12 h and then intramuscularly injected into the juvenile orange-spotted grouper. Treatment of infected tissue filtrates with anti-recombinant protein serum resulted in a significantly lower mortality of fish (Group C1, mortality of 18.18%), compared with the fish (Group C2, mortality of 40%) which received infected tissue filtrates treated with control serum. Results implied the potential use of the capsid protein in immunization against OGNNV.

  15. Perceptions of White Spot Lesions During Fixed Orthodontic Treatment of Patients, Parents, Orthodontists and Dentists%不同人群对正畸固定矫治中白垩斑的认知调查

    林佳强; 程晶晶; 毛美清; 林非欧

    2014-01-01

    Objective To assess the perceptions of patients , parents, orthodontists and general dentists on the development of white spot lesions during fixed orthodontic treatment , to prevent the development of white spot lesions and provide a reference for doctor -patient communication .Methods A survey about the perceptions of white spot lesions within orthodontic patients ( n =323 ) , parents ( n =295 ) , orthodontists (n=63) and dentists (n=117) was carried out, then analyzed.Results Four surveyed populations indicated that white spot lesions detracted the appearance of teeth .The major responsibility for the prevention of white spot lesions was owed to the patient , but not the general dentists .There is significant difference as regards the treatment of white spot lesions between dental professionals and laypeople ( P<0.01 ) .Conclusions Four populations had similar perception that white spot lesions detracted the appearance of teeth and the major responsibility for the prevention of white spot lesions was due to the patient .The patients , parents , orthodontists , and general dentists should strengthen the communication , eliminate the doctor-patient contradiction hidden dangers .%目的:调查正畸患者、患者家长、正畸医生和全科牙医对固定正畸矫治中白垩斑的认知度,为正畸治疗中白垩斑的防治及医患沟通提供参考。方法对323名正畸患者、295名患者家长、63名正畸医生及117名全科牙医进行白垩斑认知度的问卷调查,对结果进行统计分析。结果4组人群均认为白垩斑使矫治后牙齿的美观度降低,患者要对白垩斑的发生负最主要责任,患者和家长与正畸医师和牙医对白垩斑的治疗看法迥异。结论不同人群均认为白垩斑降低了牙齿的美观度,其主要责任在于患者自身,医患双方应当加强沟通,消除医患矛盾隐患。

  16. Oral Administration of CpG ODNs andYarrowia lipolytica Harboring VP28 in Farm Ponds to Protect Shrimp From White Spot Syndrome Virus Infection%自然养殖水体投喂CpG ODN和表面展示VP28的解脂耶罗维亚酵母对凡纳滨对虾(Litopenaeus vannamei)抗WSSV的免疫保护作用

    衣启麟; 刘瑞; 孙瑞; 王玲玲; 刘聪辉; 迟振明; 宋林生

    2015-01-01

    Frequent outbreaks of viral and bacterial diseases have caused severe economic loss in shrimp farming that has been an important part in aquaculture. The white spot syndrome is one of the most harmful diseases in shrimp culture. It is crucial to develop novel strategies that efficiently protect shrimp against WSSV. In this study, we performed oral administration of diet supplemented with CpG ODNs prepared with large-scale fermentation andYarrowia lipolytica harboring VP28 (VP28-yl) to farmedLitopenaeus vannamei. After 30-day feeding we challenged the shrimp with WSSV to evaluate the protective effects of CpG ODN and VP28-yl. Shrimp fed with CpG ODN displayed a relative weight gain of (65.8±7.8)% (P<0.05), indicating that CpG ODN might promote the growth of shrimp. After the WSSV challenge, the copies of WSSV in shrimp fed with CpG ODN and VP28-yl were both significantly less compared to the control (P<0.05), and the RPS were 26.7% and 36.7% respectively. The respiratory burst level of shrimp fed with CpG ODN was increased after feeding trial and WSSV challenge. After the WSSV challenge, there was a gradual increase in the apoptosis of haemocytes in the control shrimp. In the contrast, the apoptosis of haemocytes in shrimp fed with VP28-yl was significantly suppressed. Moreover, the expression ofSTAT mRNA in shrimp fed with CpG ODN and VP28-yl was significantly increased post the WSSV challenge (P<0.05), and there was an opposite trend in the control group (P<0.05). These results suggested that oral administration of CpG ODN and VP28-yl could enhance the antiviral immunity against WSSV, and thus help control viral diseases. These two immunoactive substances could be potentially new therapies in shrimp aquaculture industry.%用添加CpG寡聚核苷酸(CpG ODN)和表面展示VP28的解脂耶罗维亚酵母(VP28-yl)的饵料投喂凡纳滨对虾,进行田间中试实验.投喂30 d后进行WSSV感染实验,评估其对凡纳滨对虾的免疫保护作

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

    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.

  18. Results of calibrations of the NOAA-11 AVHRR made by reference to calibrated SPOT imagery at White Sands, N.M

    Nianzeng, Che; Grant, Barbara G.; Flittner, David E.; Slater, Philip N.; Biggar, Stuart F.; Jackson, Ray D.; Moran, M. S.

    1991-01-01

    The calibration method reported here makes use of the reflectances of several large, uniform areas determined from calibrated and atmospherically corrected SPOT Haute Resolution Visible (HRV) scenes of White Sands, New Mexico. These reflectances were used to predict the radiances in the first two channels of the NOAA-11 Advanced Very High Resolution Radiometer (AVHRR). The digital counts in the AVHRR image corresponding to these known reflectance areas were determined by the use of two image registration techniques. The plots of digital counts versus pixel radiance provided the calibration gains and offsets for the AVHRR. A reduction in the gains of 4 and 13 percent in channels 1 and 2 respectively was found during the period 1988-11-19 to 1990-6-21. An error budget is presented for the method and is extended to the case of cross-calibrating sensors on the same orbital platform in the Earth Observing System (EOS) era.

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

    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.

  20. Characterization of White Corrosion Spots on Souvenir Silver Coin%银纪念币表面滋生白斑机理研究

    康今哲; 于宏

    2011-01-01

    采用光学显微镜、扫描电镜、电子探针、X射线衍射、XPS等方法对银质纪念币在大气中放置出现的白斑的成份、结构进行了宏观和微观的分析检查,并结合湿热加速试验及银币生产现场情况,综合分析白斑的生成机理.结果表明:在湿润环境中,Cl-、O2、H2S是导致银币表面产生白斑的主要物质;银币表面白斑腐蚀产物主要由AgCl、Ag2O和Ag2S构成,腐蚀产物的颜色并不随腐蚀白斑长大而改变;银币表面存在的供氧差异腐蚀电池是银币表面白斑形核、长大的主要诱因.%Morphology and chemical composition of the corrosion spots on the surface of souvenir silver coins were examined by optical microscope(OM), scanning electron microscope(SEM), electron microprobe (EMPA), X- ray photoelectron spectroscopy(XPS), and X-ray diffraction(XRD).It was found that the silver was sensitive to the presence of Cl- 、 O2 and H2S in humid atmosphere.XPS and XRD analytical results showed that the white spots were consisted of AgC1、 Ag2O and Ag2S. The white tarnished complex on the surface of silver was a series of electrochemical process that required the presence of chloride, oxygen and sulfur.

  1. In vitro quantification of white spot enamel lesions adjacent to fixed orthodontic appliances using quantitative light-induced fluorescence and DIAGNOdent.

    Aljehani, Abdulaziz; Tranaeus, Sofia; Forsberg, Carl-Magnus; Angmar-Månsson, Birgit; Shi, Xie-Qi

    2004-12-01

    The aims of this in vitro study were 2-fold: 1) to evaluate two fluorescence methods (DIAGNOdent and QLF (quantitative light-induced fluorescence)) for quantification of white spot lesions adjacent to fixed orthodontic appliances; and 2) to determine the inter-observer agreement of the DIAGNOdent and QLF methods for quantification of incipient enamel lesions adjacent to fixed orthodontic appliances. Forty-one premolar teeth with visually sound smooth surfaces or visually white spot enamel lesions were included in the study. Orthodontic brackets were fixed adjacent to the lesions, thus simulating the position of fixed appliances during orthodontic treatment. All teeth were measured using both the DIAGNOdent and QLF methods. Of the 41 teeth, 20 smooth surfaces were randomly selected and analyzed by 4 operators using both DIAGNOdent and QLF. The teeth were sectioned into 300-microm-thick slices using a water-cooled diamond saw and the slices manually ground to 80-100 microm thickness. Histopathology and transverse microradiography were performed to provide the gold standards for verification of lesion depth and mineral loss, respectively. The Spearman rank correlation coefficients between lesion depth determined by histopathology and the DIAGNOdent and QLF were 0.76 and 0.82, respectively, whereas the Pearson correlation coefficients between mineral loss and the two methods were 0.64 and 0.84, respectively. Inter-observer agreement was found to be 0.80 and 0.93 for DIAGNOdent and QLF, respectively. In conclusion, QLF may be a suitable method for quantifying incipient carious lesions adjacent to fixed orthodontic appliances.

  2. Dried-blood spots: a cost-effective field method for the detection of Chikungunya virus circulation in remote areas.

    Soa Fy Andriamandimby

    Full Text Available BACKGROUND: In 2005, there were outbreaks of febrile polyarthritis due to Chikungunya virus (CHIKV in the Comoros Islands. CHIKV then spread to other islands in the Indian Ocean: La Réunion, Mauritius, Seychelles and Madagascar. These outbreaks revealed the lack of surveillance and preparedness of Madagascar and other countries. Thus, it was decided in 2007 to establish a syndrome-based surveillance network to monitor dengue-like illness. OBJECTIVE: This study aims to evaluate the use of capillary blood samples blotted on filter papers for molecular diagnosis of CHIKV infection. Venous blood samples can be difficult to obtain and the shipment of serum in appropriate temperature conditions is too costly for most developing countries. METHODOLOGY AND PRINCIPAL FINDINGS: Venous blood and dried-blood blotted on filter paper (DBFP were collected during the last CHIKV outbreak in Madagascar (2010 and as part of our routine surveillance of dengue-like illness. All samples were tested by real-time RT-PCR and results with serum and DBFP samples were compared for each patient. The sensitivity and specificity of tests performed with DBFP, relative to those with venous samples (defined as 100% were 93.1% (95% CI:[84.7-97.7] and 94.4% (95% CI:[88.3-97.7], respectively. The Kappa coefficient 0.87 (95% CI:[0.80-0.94] was excellent. CONCLUSION: This study shows that DBFP specimens can be used as a cost-effective alternative sampling method for the surveillance and monitoring of CHIKV circulation and emergence in developing countries, and probably also for other arboviruses. The loss of sensitivity is insignificant and involved a very small number of patients, all with low viral loads. Whether viruses can be isolated from dried blood spots remains to be determined.

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

    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.

  4. 浸渍胶膜纸饰面刨花板板面干花刍议%Discussion on Surface White Spots of Particleboard Coated with Melamine-impregnated Paper

    刘光新; 王世伟

    2016-01-01

    Discussed the causes for the white spots on the surface of particleboard coated with melamine impregnated paper and proposed solutions.%介绍浸渍胶膜纸饰面刨花板板面干花产生的原因及消除其板面干花的措施。

  5. 渗透性树脂治疗牙釉质白垩斑的研究进展%The Treatment of White Spot Lesion of Enamel Using the Resin Infiltration

    陈洪(综述); 何静(审校)

    2016-01-01

    White spot lesion is an early clinical manifestation of enamel caries .It is one of common com-plications during the orthodontic treatment.It affects the appearance of the teeth.The resin infiltration is a new kind of resin with low viscosity,which is an effective and non-invasive method of treating the white spot lesion of enamel.In recent years,many scholars pay attention to it and are spreading it in clinical application . Here is to discuss the reason of formation of white spot lesion and the properties of this infiltration resin ,and analyze the clinical effect and the theoretical basis of this resin treating the white spot lesion ,and validate the effectiveness of this resin for treating the white spot lesion .%白垩斑是釉质龋早期的临床表现,是正畸治疗中常见的并发症之一,影响美观。渗透性树脂作为一种新型的树脂具有低黏性,是近年来治疗早期釉质龋白垩斑有效且无创的新方法。该法受到许多学者的关注和口腔临床医师的推广运用。该文通过论述白垩斑发生的原因和渗透性树脂的特性,阐述了渗透性树脂治疗白垩斑的理论依据,并分析渗透性树脂的临床疗效,证实了渗透性树脂治疗白垩斑的有效性。

  6. West nile virus in American white pelican chicks: transmission, immunity, and survival

    Sovada, Marsha A.; Pietz, Pamela J.; Hofmeister, Erik K.; Bartos, Alisa J.

    2013-01-01

    West Nile virus (WNV) causes significant mortality of American White Pelican chicks at northern plains colonies. We tested oropharyngeal/cloacal swabs from moribund chicks for shed WNV. Such shedding could enable chick-to-chick transmission and help explain why WNV spreads rapidly in colonies. WNV was detected on swabs from 11% of chicks in 2006 and 52% of chicks in 2007; however, viral titers were low. Before onset of WNV mortality, we tested blood from maternal antibodies. Among near-fledged chicks, 41% tested positive for anti-WNV antibodies, indicating that they survived infection. Among years and colonies, cumulative incidence of WNV in chicks varied from 28% to 81%, whereas the proportion of chicks surviving WNV (i.e., seropositive) was 64–75%. Our data revealed that WNV kills chicks that likely would fledge in the absence of WNV, that infection of chicks is pervasive, and that significant numbers of chicks survive infection.

  7. Influence of boat noises on escape behaviour of white-spotted eagle ray Aetobatus ocellatus at Moorea Island (French Polynesia).

    Berthe, Cecile; Lecchini, David

    2016-02-01

    The present study tested different sounds that could disturb eagle rays (Aetobatus ocellatus) during their foraging activities at Moorea, French Polynesia. Results showed that artificial white sound and single-frequency tones (40 Hz, 600 Hz or 1 kHz) did not have an effect on rays (at least 90% of rays continued to forage over sand), while playbacks of boat motor sound significantly disturbed rays during foraging activity (60% exhibited an escape behaviour). Overall, our study highlighted the negative effect of boat noises on the foraging activity of eagle rays. These noises produced by boat traffic could, however, have some positive effects for marine aquaculture if they could be used as a deterrent to repel the eagle rays, main predators of the pearl oysters.

  8. Comparison of immune response of Pacific white shrimp, Litopenaeus vannamei, after multiple and single infections with WSSV and Vibrio anguillarum

    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 [1]. For better understanding the defense mechanism against white spot syndrome virus (WSSV...

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

    Feijó, Rubens Galdino; Maggioni, Rodrigo; Cunha 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.

  10. The incidence of caries and white spot lesions in orthodontically treated adolescents with a comprehensive caries prophylactic regimen--a prospective study.

    Hadler-Olsen, Sigurd; Sandvik, Kristin; El-Agroudi, Mohammad A; Øgaard, Bjørn

    2012-10-01

    The aim of the study was to assess the effect of a comprehensive prophylactic regimen in reducing the incidence of white spot lesions (WSL) and caries during orthodontic treatment. Eighty consecutive patients, scheduled for fixed appliance treatment in both jaws were compared with a non-orthodontic matched-control group. The oral hygiene regimen consisted of brushing two to three times daily, flossing, fluoride rinse, and plaque disclosing tablets. Patients were requested to avoid carbonated soft drinks/acidic juices and candies. The WSL index of Gorelick et al. (Gorelick L, Geiger A M, Gwinnett A J 1982 Incidence of white spot formation after bonding and banding. American Journal of Orthodontics 81: 93-98) was used. Caries were scored according to Amarante et al. (Amarante E, Raadal M, Espelid I 1998 Impact of diagnostic criteria on the prevalence of dental caries in Norwegian children aged 5, 12 and 18 years. Community Dental Oral Epidemiology 26: 87-94). We collected data from all finished cases. It comprised 40 subjects in the orthodontic group (mean age: 13.6 years, range: 12-16 years) and 40 matched controls. The average treatment time was 18 months (range: 9-25 months). The mean increase in WSL index in the orthodontic group was 1.9 and 0.4 in the control group (P = 0.001). The mean increase in dentine caries was 0.5 lesions and 0.7 lesions in the in the orthodontic group and control group, respectively (P = 0.62). Twenty-three per cent of treated patients showed good compliance, 68 per cent moderate compliance, and 9 per cent poor compliance. The mean increase in WSL was 1.0, 1.4, and 3.3 in the good, moderate, and poor compliance group, respectively (P = 0.155). Orthodontically treated patients have significantly higher risk for developing WSL than untreated patients, while there is no difference with respect to development of new dentinal caries lesions. This study showed that a possible relationship between compliance and WSL development existed.

  11. Assessment of the risk of White Sturgeon to become infected and potential carriers of infectious pancreatic necrosis virus.

    LaPatra, Scott E; Mead, Sherry

    2013-12-01

    Little scientific information is available to assess whether White Sturgeon Acipenser transmontanus can become infected and potential carriers of infectious pancreatic necrosis virus (IPNV). To assess this risk, monitoring results of adult and progeny White Sturgeon were examined from waters historically stocked with salmonid fish known to be IPNV carriers. From 1999 through 2004 White Sturgeon from a total of 30 separate families whose parentage came from waters historically stocked with IPNV carrier fish were tested. Duplicate groups of 25 juvenile Snake River White Sturgeon were waterborne exposed to 1.0×10(4) 50% tissue culture infective dose (TCID50)/mL of water for 1 h and an additional group was injected intraperitoneally with 1.0×10(5) TCID50/fish. A negative control group was handled similarly but was not exposed to the virus. No morbidity was detected in any of the treatment groups or the negative control. At 34, 40, 47, and 54 d postexposure to IPNV, virus reisolation was attempted on five fish from each group, and an additional five fish from each group were examined for histological changes consistent with an IPNV infection. At 34 and 40 d postinjection with IPNV, 20% (one of five) of the fish tested positive for the virus per sample interval; however, fish from groups that were waterborne-exposed to IPNV were all negative. At 47 and 54 d after exposure or injection with IPNV an additional five fish from each group were tested at each sample interval and all results were negative. Histological analysis of target tissue obtained from five fish per group at 34 and 54 d postinfection also failed to detect any consistent change associated with an IPNV infection. These results suggest that the risk of White Sturgeon to become infected and develop into potential carriers of IPNV is negligible.

  12. 中外白糖期现货价格波动及共生关系研究%Study of White Sugar Future and Spot Price's Fluctuations and Symbiotic Relationships at Home and Abroad

    袁庆禄

    2012-01-01

    构建GARCH模型,揭示糖11期指、郑糖期指和柳糖现指的波动规律及共生关系。估计结果表明:三种指数均具有尖峰厚尾特征;糖11期指和郑糖期指均存在杠杆效应;国内郑糖期指和柳糖现指表现出明显的共生关系,中外白糖指数之间的共牛关系不明显。%This paper established GARCH model, and comparatively analysed the fluctuation characteristics and symbiotic relationships of ICE sugar No.l 1 tuture index, Zhengzhou white sugar future index and Liuzhou white sugar spot index. The results showed that three indices had the characteristics of higher peak and fat tail. Both ICE sugar No. 11 future index and Zhengzhou white sugar future index had asymmetry effect. There was a symbiotic relationship between Zhengzhou white sugar future index and Liuzhou white sugar spot index obviously, and wasn't a symbiotic relationship between domestie index and foreign index.

  13. Efficiency of resin infiltration versus fluride varnish for treatment of post-orthodontic white spot lesions%渗透性树脂与氟漆治疗正畸后白垩斑的疗效比较

    王璐; 简洁; 陆慧芳

    2013-01-01

      背景:牙釉质脱矿所引起的白垩斑是正畸治疗后常见的不良反应之一。近年来,渗透性树脂在对正畸后白垩斑的治疗中取得了很好的效果。但是渗透性树脂与常规治疗方法氟漆对正畸后白垩斑的疗效有何差别尚未见报道。目的:比较渗透性树脂与氟漆治疗正畸后白垩斑的疗效。方法:选取29例正畸结束后白垩斑患者,共70颗病患牙,分别采取渗透性树脂和氟漆治疗;收集患者白垩斑治疗前以及治疗6个月后的正面口内照片,分别经口腔医生与非口腔专业的大学生对白垩斑改善程度进行评估。结果与结论:除1例氟漆治疗患者失访外,其余患者均按期完成了复诊。口腔医生和非口腔专业的大学生评分结果显示,两组治疗后白垩斑均显著改善(P<0.05);且渗透性树脂对于正畸治疗后白垩斑的治疗效果要优于氟漆治疗。可见渗透性树脂治疗具有复诊次数少,治疗效果好等优点,是治疗正畸后白垩斑的较理想方法。%BACKGROUND:White spot lesions caused by enamel demineralization are a common side effect of orthodontic treatment. Resin infiltration treatment for white spot lesions has obtained favorable effects, but there is no report on the difference between resin infiltration and fluoride varnish treatment for post-orthodontic white spot lesions. OBJECTIVE:To compare the efficiency of resin infiltration and fluride varnish for treatment of post-orthodontic white spot lesions. METHODS:Twenty-nine patients with post-orthodontic white spot lesions were randomly divided into two groups:one group accepted the resin infiltration treatment and the other group accepted fluoride varnish treatment. Frontal intraoral photos before treatment and 6 months after treatment were col ected for study. The difference in efficiency of resin infiltration and fluride varnish for treatment of post-orthodontic white spot lesion was analyzed

  14. Prevalence of white spot lesion in a section of Indian population undergoing fixed orthodontic treatment: An in vivo assessment using the visual International Caries Detection and Assessment System II criteria

    Nandikolla Sagarika; Sundaramoorthy Suchindran; S C Loganathan; Velayutham Gopikrishna

    2012-01-01

    Background: Enamel demineralization resulting in White Spot Lesion (WSL) is one of the most common disease outcome associated with fixed orthodontic therapy with high prevalence rates in western countries. However, no such studies have been done on Indian population. Aim: The aim of this study was to evaluate the prevalence of WSLs in a section of urban Indian population between the age group of 12-20 years undergoing fixed orthodontic therapy for a period of 12-15 months. Materials an...

  15. Inactivation of Anisakis simplex L3 in the flesh of white spotted conger (Conger myriaster) by high hydrostatic pressure and its effect on quality.

    Lee, Ki-Hoon; Park, Shin Young; Ha, Sang-Do

    2016-06-01

    Koreans consume much seafood; the country is surrounded on the east, west and south by the sea. Koreans have eaten raw sashimi for a long time. However, a concern in the raw sea food industry is that the parasitic nematode Anisakis simplex L3 occurs naturally in marine fish. Thus, the fishery industry needs a non-thermal processing method. High hydrostatic pressure (HPP) has been demonstrated to be effective. White spotted conger flesh containing 20 live larvae was exposed to different pressures (150 and 200 MPa for 1 and 5 min; 250 and 300 MPa each for 1 min). The viability of A. simplex L3 was significantly (p controls or that treated at 200 MPa for 5 min. No significant changes (p > 0.05) in any of the Hunter colour ('L', 'a' and 'b') values were found after HPP at 200 MPa for 5 min. The fresh treated at 300 MPa for 1 min scored quality) of flavour, texture and overall acceptability in untrained sensory evaluation using a seven-point hedonic scale. However, the flesh treated at 200 MPa for 5 min scored > 5.0 ('like') for all sensory parameters. This study suggested that HPP at 200 MPa for 5 min could potentially be used for the inactivation of A. simplex L3 in raw fishery food products without any concomitant changes in their colour or sensory qualities.

  16. 白斑综合征病毒(White spot syndrome virus)的形态和分子生物学

    王吉桥; 姜玉声; 张建

    2008-01-01

    白斑综合征病毒(White spot syndrome virus,WSSV)自1992年首次发现以来,已成为威胁对虾养殖业的最主要病原。10年中它迅速蔓延至世界各养虾区,造成直接经济损失达700多亿美元。WSSV属线头病毒科Nimaviridae,完整包膜的病毒体(粒子)长210~380nm,最宽处70—167nm,病毒体末端具尾状附器(tail-like appendage)。WSSV被膜厚6—7nm,为脂质三层膜结构,由不透明电子层分隔成两层透明层。核衣壳位于膜内,为弹簧体型结构,由直径10nm的球形蛋白亚基构成。WSSV基因组为环状双链DNA分子,A+T含量占59%,分布均匀,发生分为六个时期。我国WSSV基因组有9.63kbp的不稳定区,依寄主的不同发生了不同程度的却失,使致毒力发生了变化。

  17. Research on the White Spots Appeared on Gold Ornaments' Wearing Process%金饰品在佩戴过程中出现白色斑点的问题探究

    那勃; 曹维宇; 孟建华; 崔建军

    2015-01-01

    The white spots appeared on gold ornaments in wearing process were studied. In the experiment, gold pieces were soaked into mercury ion solutions with different concentrations. The results showed that mercury ion alone can not change gold color into white; under the conditions of adding reduction agent and the concentration of mercury ion reaching a certain value, the white spots appeared on gold pieces. The relationship between concentration of mercury ion and white changing degree of gold pieces was studied.%针对金饰品在佩戴过程中形成白色斑点的问题进行了研究.将金片浸泡到不同浓度的汞离子溶液中,实验结果表明,单纯的汞离子无法令黄金变白;在加入还原剂,且汞离子达到一定浓度条件下,金片出现明显变色现象;研究了汞离子浓度和金片变白程度的关系.

  18. Diagnosis of nervous necrosis virus in orange-spotted grouper, Epinephelus coioides, by a rapid and convenient RT-PCR method

    MU Yinnan; LIN Kebing; CHEN Xinhua; AO Jingqun

    2013-01-01

    Viral nervous necrosis (VNN) causes high mortality in marine fish, especially in the grouper, worldwide and in China. Since there is no effective vaccine or drug to deal with VNN, early detection and prevention is important to block its outbreak. In this study, a reverse transcription-polymerase chain reaction (RT-PCR) was developed for the rapid, convenient, and sensitive detection of the VNN pathogen, nervous necro-sis virus (NNV ), in the grouper. The whole process was completed within 3.5 h from the RNA extraction to PCR product visualization. The detection limit of this method was 200 copies of NNV RNA standard, which corresponded to 200 copies of virus particles. This RT-PCR method was specific to the NNV detection with no cross-reactivity to other fish viral disease pathogens, such as infectious pancreatic necrosis virus (IPNV ), infectious hematopoietic necrosis virus (IHNV ), spring viraemia of carp virus (SVCV ), epizootic haematopoietic necrosis virus (EHNV ), and large yellow croaker iridovirus (LYCIV ). With this method, the orange-spotted grouper (Epinephelus coioides) fry from hatcheries with or without incidence of the VN-N epidemic in Fujian Province were detected. The results showed that all or 93%of the fry from the two hatcheries with incidence of the epidemic were diagnosed as positive, while 40%or 25%of fry from the t-wo hatcheries without the VNN epidemic were also detected as NNV positive, indicating that this RT-PCR method can be used for rapid, sensitive detection of NNV infection and applied in the VNN epidemic alert.

  19. BEBERAPA METODE STRESSING UNTUK MENGINDUKSI PERKEMBANGAN WHITE SPOT SYNDROME VIRUS (WSSV PADA BENUR UDANG WINDU (Penaeus monodon

    Muliani Muliani

    2012-12-01

    Full Text Available Penelitian ini bertujuan untuk mengetahui beberapa model metode stressing yang dapat menginduksi perkembangan WSSV pada benur, sehingga dapat terdeteksi dengan PCR konvensional. Penelitian dilakukan di Laboratorium Basah Balai Penelitian dan Pengembangan Budidaya Air Payau (BPPBAP. Wadah yang digunakan adalah kontainer plastik bervolume 40 L diisi dengan air laut sebanyak 30 L dan dilengkapi dengan aerasi kecuali pada perlakuan oksigen. Setiap wadah ditebari benur (PL-12 sebanyak 500 ekor yang diambil dari panti perbenihan komersil di Kabupaten Barru yang sebelumnya telah diketahui negatif WSSV. Perlakuan yang dicobakan adalah: (A stressing dengan suhu yaitu: 10oC±2oC, 28oC±2oC, dan 40oC±2oC; (B stressing dengan salinitas yaitu: 5 ppt, 28 ppt, dan 51 ppt; (C stressing dengan oksigen yaitu: 1 mg/L, 3 mg/L, dan 5 mg/L; (D stressing dengan pH yaitu: pH 5, pH 6, dan pH 7; dan (E stressing dengan formalin yaitu: 100 mg/L, 200 mg/L, dan 300 mg/L. Sampling dilakukan 1 jam, 3 jam, dan 5 jam setelah perendaman untuk pengecekan WSSV dengan mengambil contoh benur sebanyak 50-100 ekor/wadah lalu dicek dengan PCR untuk WSSV-nya. Hasil penelitian menunjukkan bahwa stressing dengan oksigen (1 mg/L, 3 mg/L, dan 5 mg/L memberikan hasil yang terbaik dalam memicu perkembangan WSSV pada benur, disusul dengan suhu (suhu 10oC dan 40oC, pH 5; kemudian penggunaan formalin, dan salinitas. Hasil PCR menunjukkan bahwa benur yang awalnya  negatif menjadi positif dengan kategori ringan hingga berat. Semakin lama waktu perendaman maka semakin memicu perkembangan WSSV dalam badan benur.

  20. [Identification of Host Factors Interacting with the Movement Protein of Apple Chlorotic Leaf Spot Virus by Yeast Two-Hybrid System].

    He, Yikun; Zhong, Min; Zhang, Yu; Wang, Yanan; Cao, Keqiang

    2015-03-01

    In order to identify host factors which interact with the movement protein (MP) of Apple chlorotic leaf spot virus (ACLSV), ACLSV MP was cloned into the bait vector pGBKT7 and used to screen a cDNA library of Malus sylvestris cv. R12740-7A, which had previously been constructed by yeast two-hybrid sequencing transformation. The protein functions of the identified host factors were determined according to their gene annotations in GenBank. The result showed that the bait plasmid pGBKT7-MP showed no virulence or self-activating effect on yeast strain Y2H Gold. Sixty-nine interactor proteins were identified, which were divided into the following 10 classes according to their described functions: hydrolases; pathogenesis-related proteins; DNA binding proteins; phosphatases; ligases; proteins with catalytic activity; phenylalanine ammonialyases; peroxidases; NAD binding proteins; and proteins of unknown function. Bioinformatic analysis of gene homology suggested that phosphatases, pathogenesis-related proteins and glyceraldehyde-3-phosphate dehydrogenase A may play an important role in the interaction between virus and host. This study may provide a theoretical basis for the further study of viral pathogenesis and virus-host interaction mechanisms.

  1. Sequence diversity of the nucleoprotein gene of iris yellow spot virus (genus Tospovirus, family Bunyaviridae) isolates from the western region of the United States.

    Pappu, H R; du Toit, L J; Schwartz, H F; Mohan, S K

    2006-05-01

    Iris yellow spot virus (IYSV), a tentative virus species in the genus Tospovirus and family Bunyaviridae, is considered a rapidly emerging threat to onion production in the western United States (US). The present study was undertaken to determine the sequence diversity of IYSV isolates from infected onion plants grown in California, Colorado, Idaho, Oregon, Utah and Washington. Using primers derived from the small RNA of IYSV, the complete sequence of the nucleoprotein (NP) gene of each isolate was determined and the sequences compared. In addition, a shallot isolate of IYSV from Washington was included in the study. The US isolates of IYSV shared a high degree of sequence identity (95 to 99%) with one another and to previously reported isolates. Phylogenetic analyses showed that with the exception of one isolate from central Oregon and one isolate from California, all the onion and shallot isolates from the western US clustered together. This cluster also included onion and lisianthus isolates from Japan. A second distinct cluster consisted of isolates from Australia (onion), Brazil (onion), Israel (lisianthus), Japan (alstroemeria), The Netherlands (iris) and Slovenia (leek). The IYSV isolates evaluated in this study appear to represent two distinct groups, one of which largely represents isolates from the western US. Understanding of the population structure of IYSV would potentially provide insights into the molecular epidemiology of this virus.

  2. 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

    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.

  3. Empirical study on relationship between futures price and spot price of white sugar in China%我国白糖期货价格和现货价格关系的实证研究∗

    罗丹程; 黄月; 王添钰

    2016-01-01

    选取2006年3月1日—2016年3月31日的白糖期货和现货市场数据,通过相关性检验、ADF单位根检验、VAR模型、Johansen协整检验和格兰杰因果检验等分析方法,指出白糖期货价格与其现货价格存在长期均衡关系,且两者高度相关;白糖期货价格对未来白糖现货价格具有引导作用,但这个引导作用并不是相互的。同时,提出建立完善的期货市场制度及市场环境的建议,以充分发挥白糖期货市场的价格发现功能,形成更有效、更公平的白糖价格体系。%The market data of the futures prices and spot prices of white sugar are sampled from March 1st, 2006 to March 31st, 2016. Such analysis methods are used as correlation test, ADF unit root test, VAR model, Johansen co-integration test, and Grainger causality test. It is proposed that the futures prices and spot prices of white sugar have the relationship in the long-run equilibrium, and are highly correlated; and that futures price of white sugar has a guiding role on the spot price of white sugar in the future, but not vice versa. Suggestions are proposed concerning the establishment of perfect futures market system and market environment, so as to give full play to the price discovery function of white sugar futures market, and to form a more effective and equitable system of white sugar prices.

  4. Age Spots

    ... How to Choose the Best Skin Care Products Age Spots Treatment Options Learn more about treatment options ... or black, flat patches of skin. Why treat age spots Physical, emotional and social reasons for treating ...

  5. Virus Survey of Commercial Cowpea Fields in the U. S. and Evaluation of the Core of the USDA Cowpea Germplasm Collection for Resistance to Tomato Spotted Wilt Virus

    Cowpea (Vigna unguiculata) is an important vegetable crop in the southern United States. Cowpea plants are susceptible to over 20 viruses and many of them are seed-borne. A survey was carried out to evaluate the current virus status in commercial cowpea fields in five States (MO, NM, SC, TN and TX...

  6. Effect of resin infiltration treatment on the colour of white spot lesions%渗透树脂治疗对釉质白斑样病损颜色的影响

    赵晓一; 高学军

    2014-01-01

    目的:研究渗透树脂治疗对釉质白斑样病损颜色的影响。方法选择人磨牙60颗,树脂包埋,以牙尖方向作为观察面,每个样本打磨出2个釉质小平面,随机分为A、B点。60颗样本牙随机分为第1、2、3组,每组20颗,经人工龋脱矿液分别脱矿24、48、72h;对各样本的A点进行渗透树脂处理,B点行0.1%氟化钠涂氟处理30d,B点涂氟处理后再行渗透树脂处理。应用电子分光光度比色仪测量A、B点脱矿及处理前后的L*值。结果各组A、B点脱矿前L*值没有明显差异,脱矿后均明显增加;经渗透树脂处理后,各组A点的L*值均较脱矿后降低,第1、2组的L*值恢复到脱矿前水平,第3组L*值与第1、2组存在差异(P0.05), whereas the L* values of spots A and B were significantly increased after demineralization. The L* values of A spots recovered signifi-cantly after treatment by resin infiltration (P0.05) after fluoride treatment compared with that after demineralization. After resin infil-tration on B spots, the L* values recovered but could not reach the baseline nor the level of A spots treated by resin infiltration only. Conclusion Resin infiltration is a more effective treatment for masking white spot lesions than traditional fluoride treatment. The effect of masking white spot lesions has certain relationships with the degree of demineralization and activity of the lesion.

  7. Large-scale codon de-optimisation of the p29 replicase gene by synonymous substitutions causes a loss of infectivity of melon necrotic spot virus.

    Usami, Atsushi; Mochizuki, Tomofumi; Tsuda, Shinya; Ohki, Satoshi T

    2013-09-01

    The effect of synonymous substitutions in the melon necrotic spot virus p29 replicase gene on viral pathogenicity was investigated. The codons in the p29 gene were replaced by the least frequently used synonymous codons in Arabidopsis thaliana or melons. Mechanical inoculation of melon with p29 variants resulted in a loss of viral infectivity when all, one-half, or one-quarter of the gene was de-optimised. The effect of the de-optimisation in one-sixth of the gene was different depending on the de-optimised region. These results demonstrate that large-scale codon bias de-optimisation without amino acid substitutions of the p29 gene alter viral infectivity.

  8. Tomato yellow spot virus, a tomato-infecting begomovirus from Brazil with a closer relationship to viruses from Sida sp., forms pseudorecombinants with begomoviruses from tomato but not from Sida.

    Andrade, E C; Manhani, G G; Alfenas, P F; Calegario, R F; Fontes, E P B; Zerbini, F M

    2006-12-01

    Geminiviruses are characterized by a circular, single-stranded DNA genome and twinned icosahedral particles. Begomoviruses (whitefly-transmitted geminiviruses) are a major constraint to crop production worldwide. In Brazil, tomato-infecting begomoviruses emerged as serious pathogens over the last 10 years, due to the introduction of a new biotype of the insect vector. Tomato yellow spot virus (ToYSV) is a newly described begomovirus originally isolated from tomato, but phylogenetically closer to viruses from Sida sp. A study was performed to determine the viability of pseudorecombinants formed between the DNA components of ToYSV and other weed- and tomato-infecting begomoviruses from Brazil. Despite its closer relationship to weed-infecting viruses, ToYSV was only capable of forming viable pseudorecombinants with tomato viruses. An infectious pseudorecombinant formed between ToYSV DNA-A and tomato crinkle leaf yellows virus (TCrLYV) DNA-B induced severe symptoms in Nicotiana benthamiana. This was attributed, at least in part, to the fact that the origins of replication of both components had identical Rep-binding sequences. However, this was not the case for another infectious pseudorecombinant formed between tomato golden mosaic virus (TGMV) DNA-A and ToYSV DNA-B, which have different Rep-binding sequences. These results reinforce the notion that pseudorecombinant formation cannot be explained solely on the basis of phylogenetic relationships and conserved iteron sequences, and suggest that the TGMV Rep protein may be more versatile in terms of recognizing heterologous DNA components than that of ToYSV.

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

    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

  10. Biological impacts of "hot-spot" mutations of hepatitis B virus X proteins are genotype B and C differentiated

    Xu Lin; Xiao Xu; Qing-Ling Huang; Yu-Qing Liu; Da-Li Zheng; Wan-Nan Chen; Jian-Yin Lin

    2005-01-01

    AIM: To investigate the biological impacts of "hot-spot"mutations on genotype B and C HBV X proteins (HBx).METHODS: Five types of"hot-spot" mutations of genotype B or C HBV X genes, which sequentially lead to the amino acid substitutions of HBx as I127T, F132Y, K130M+V131I,I127T+K130M+V131I, or K130M+V131I+F132Y, respectively,were generated by means of site-directed mutagenesis.To evaluate the anti-proliferative effects, HBx or related mutants' expression vectors were transfected separately to the Chang cells by lipofectamine, and the cells were cultured in hygromycin selective medium for 14 d, drug-resistant colonies were fixed with cold methanol, stained with Giemsa dyes and scored (increase of the colonies indicated the reduction of the anti-proliferation activity,and vice versa). Different types of HBx expression vectors were co-transfected separately with the reporter plasmid pCMVβ to Chang cells, which were lysed 48 h post-transfection and the intra-cellular β-galactosidase activities were monitored (increase of the β-galactosidase activities indicated the reduction of the transactivation activity, and vice versa). All data obtained were calculated by paired-samples t-test.RESULTS: As compared to standard genotype B HBx,mutants of I127T and I127T+K130M+V131I showed higher transactivation and anti-proliferative activities, while the mutants of F132Y, K130M+V131I, and K130M+V131I+F132Y showed lower activities. As compared to standard genotype C HBx, I127T mutant showed higher transactivation activity, while the other four types of mutants showed no differences. With regard to anti-proliferative activity,compared to standard genotype C HBx, F132Y and K130M+V131I mutants showed lower activities, and K130M+V131I +F132Y mutant, on the other hand, showed higher activity,while the mutants of I127T and I127T+K130M+V131I showed no differences.CONCLUSION: "Hot-spot" mutations affect the antiproliferation and transactivation activities of genotype B and/or C HBx, and

  11. Characterization of Imjin virus, a newly isolated hantavirus from the Ussuri white-toothed shrew (Crocidura lasiura).

    Song, Jin-Won; Kang, Hae Ji; Gu, Se Hun; Moon, Sung Sil; Bennett, Shannon N; Song, Ki-Joon; Baek, Luck Ju; Kim, Heung-Chul; O'Guinn, Monica L; Chong, Sung-Tae; Klein, Terry A; Yanagihara, Richard

    2009-06-01

    Until recently, the single known exception to the rodent-hantavirus association was Thottapalayam virus (TPMV), a long-unclassified virus isolated from the Asian house shrew (Suncus murinus). Robust gene amplification techniques have now uncovered several genetically distinct hantaviruses from shrews in widely separated geographic regions. Here, we report the characterization of a newly identified hantavirus, designated Imjin virus (MJNV), isolated from the lung tissues of Ussuri white-toothed shrews of the species Crocidura lasiura (order Soricomorpha, family Soricidae, subfamily Crocidurinae) captured near the demilitarized zone in the Republic of Korea during 2004 and 2005. Seasonal trapping revealed the highest prevalence of MJNV infection during the autumn, with evidence of infected shrews' clustering in distinct foci. Also, marked male predominance among anti-MJNV immunoglobulin G antibody-positive Ussuri shrews was found, whereas the male-to-female ratio among seronegative Ussuri shrews was near 1. Plaque reduction neutralization tests showed no cross neutralization for MJNV and rodent-borne hantaviruses but one-way cross neutralization for MJNV and TPMV. The nucleotide and deduced amino acid sequences for the different MJNV genomic segments revealed nearly the same calculated distances from hantaviruses harbored by rodents in the subfamilies Murinae, Arvicolinae, Neotominae, and Sigmodontinae. Phylogenetic analyses of full-length S, M, and L segment sequences demonstrated that MJNV shared a common ancestry with TPMV and remained in a distinct out-group, suggesting early evolutionary divergence. Studies are in progress to determine if MJNV is pathogenic for humans.

  12. Characterization of Imjin Virus, a Newly Isolated Hantavirus from the Ussuri White-Toothed Shrew (Crocidura lasiura)▿

    Song, Jin-Won; Kang, Hae Ji; Gu, Se Hun; Moon, Sung Sil; Bennett, Shannon N.; Song, Ki-Joon; Baek, Luck Ju; Kim, Heung-Chul; O'Guinn, Monica L.; Chong, Sung-Tae; Klein, Terry A.; Yanagihara, Richard

    2009-01-01

    Until recently, the single known exception to the rodent-hantavirus association was Thottapalayam virus (TPMV), a long-unclassified virus isolated from the Asian house shrew (Suncus murinus). Robust gene amplification techniques have now uncovered several genetically distinct hantaviruses from shrews in widely separated geographic regions. Here, we report the characterization of a newly identified hantavirus, designated Imjin virus (MJNV), isolated from the lung tissues of Ussuri white-toothed shrews of the species Crocidura lasiura (order Soricomorpha, family Soricidae, subfamily Crocidurinae) captured near the demilitarized zone in the Republic of Korea during 2004 and 2005. Seasonal trapping revealed the highest prevalence of MJNV infection during the autumn, with evidence of infected shrews' clustering in distinct foci. Also, marked male predominance among anti-MJNV immunoglobulin G antibody-positive Ussuri shrews was found, whereas the male-to-female ratio among seronegative Ussuri shrews was near 1. Plaque reduction neutralization tests showed no cross neutralization for MJNV and rodent-borne hantaviruses but one-way cross neutralization for MJNV and TPMV. The nucleotide and deduced amino acid sequences for the different MJNV genomic segments revealed nearly the same calculated distances from hantaviruses harbored by rodents in the subfamilies Murinae, Arvicolinae, Neotominae, and Sigmodontinae. Phylogenetic analyses of full-length S, M, and L segment sequences demonstrated that MJNV shared a common ancestry with TPMV and remained in a distinct out-group, suggesting early evolutionary divergence. Studies are in progress to determine if MJNV is pathogenic for humans. PMID:19357167

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

    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.

  14. Complete sequence of three different biotypes of tomato spotted wilt virus (wild type, tomato Sw-5 resistance-breaking and pepper Tsw resistance-breaking) from Spain.

    Debreczeni, Diana E; López, Carmelo; Aramburu, José; Darós, José Antonio; Soler, Salvador; Galipienso, Luis; Falk, Bryce W; Rubio, Luis

    2015-08-01

    Tomato spotted wilt virus (TSWV) occurs worldwide and causes production losses in many important horticultural crops such as tomato and pepper. Breeding resistant cultivars has been the most successful method so far for TSWV disease control, but only two genes have been found to confer resistance against a wide spectrum of TSWV isolates: Sw-5 in tomato and Tsw in pepper. However, TSWV resistance-breaking isolates have emerged in different countries a few years after using resistant cultivars. In this paper, we report the first complete nucleotide sequences of three Spanish TSWV isolates with different biotypes according to their abilities to overcome resistance: LL-N.05 (wild type, WT), Pujol1TL3 (Sw-5 resistance breaking, SBR) and PVR (Tsw resistance-breaking, TBR). The genome of these TSWV isolates consisted of three segments: L (8913-8914 nt), M (4752-4825 nt) and (S 2924-2961 nt). Variations in nucleotide sequences and genomic RNA lengths among the different virus biotypes are reported here. Phylogenetic analysis of the five TSWV open reading frames showed evidence of reassortment between genomic segments of LL-N.05 and Pujol1TL3, which was supported by analysis with different recombination-detecting algorithms.

  15. The movement protein (NSm) of Tomato spotted wilt virus is the avirulence determinant in the tomato Sw-5 gene-based resistance.

    Peiró, Ana; Cañizares, M Carmen; Rubio, Luis; López, Carmelo; Moriones, Enrique; Aramburu, José; Sánchez-Navarro, Jesús

    2014-10-01

    The avirulence determinant triggering the resistance conferred by the tomato gene Sw-5 against Tomato spotted wilt virus (TSWV) is still unresolved. Sequence comparison showed two substitutions (C118Y and T120N) in the movement protein NSm present only in TSWV resistance-breaking (RB) isolates. In this work, transient expression of NSm of three TSWV isolates [RB1 (T120N), RB2 (C118Y) and non-resistance-breaking (NRB)] in Nicotiana benthamiana expressing Sw-5 showed a hypersensitive response (HR) only with NRB. Exchange of the movement protein of Alfalfa mosaic virus (AMV) with NSm supported cell-to-cell and systemic transport of the chimeric AMV RNAs into N. tabacum with or without Sw-5, except for the constructs with NBR when Sw-5 was expressed, although RB2 showed reduced cell-to-cell transport. Mutational analysis revealed that N120 was sufficient to avoid the HR, but the substitution V130I was required for systemic transport. Finally, co-inoculation of RB and NRB AMV chimeric constructs showed different prevalence of RB or NBR depending on the presence or absence of Sw-5. These results indicate that NSm is the avirulence determinant for Sw-5 resistance, and mutations C118Y and T120N are responsible for resistance breakdown and have a fitness penalty in the context of the heterologous AMV system.

  16. 白车身电阻点焊质量控制技术%Quality measurements for resistance spot-weld and control technologies in body-in-white

    张勇; 付玉生

    2012-01-01

    Several technology methods and their applications for spot-weld quality measurement and control were briefly described in this article.The single channel ultra-sonic back wave of different characteristic on weld-spots is list.And latest matrix ultra-sonic aenor information, dynamic resistance self-adaption control technology are described in the article.The application and development Irend of ultra-sonic measure technology and dynamic resistance self-adaption control technology in quality assurance system of spot-weld in Body-in-White also introduced.%概述了白车身电阻点焊质量检测控制的几种技术手段,列举了常见的不同特征焊点的单通道超声波回馈波形,介绍目前最新的矩阵式超声波传感器的特点和动态电阻自适应控制技术工作原理,以及超声波检测技术和动态电阻自适应控制技术在车身电阻点焊质量保证系统中的应用情况及其趋势.

  17. Effects of fixed appliances during orthodontic treatment on the incidence of white spot lesions of enamel inviw.%固定正畸矫治器对牙釉质龋白斑患病率的影响

    王丽芬; 黄志刚; 张霞

    2011-01-01

    目的:研究固定正畸矫治器治疗 6 个月和 12 个月时牙釉质龋白斑的患病率,为有效控制牙釉质的早期脱矿提供依据.方法:将 114 名进行固定正畸矫治病例分为三组:6 月组、12 月组和空白对照组,检查各组患者靠近托槽龈方牙面的牙釉质龋白斑患病率和龋白斑指数.对各组间牙釉质龋白斑患病率和龋白斑指数进行单因素方差分析,Logistic 回归分析检验治疗时间和性别对牙釉质龋白斑患病率的多重影响.结果:76%的男性患者和 24%的女性患者出现了牙釉质龋白斑,其患病率的差异具有统计学意义(P0.05),但与对照组间的差异均具有统计学意义(P<0.05).结论:患者行固定矫治器治疗时牙釉质龋白斑的患病率在最初 6 个月时急剧上升,因此,临床医生应注意在患者进行固定矫治器治疗的最初数月应密切关注其口腔卫生状况,必要时应采取措施防止牙釉质的脱矿.%Objective: The aim of this in vivo study was to investigate the effects of fixed appliances during orthodontic treatment on the incidence of white spot lesions of enamel. Method: 114 patients who were examined for the presence of enamel demineralization were divided into three groups: 6-month group, 12-month group and the control group. Tooth surfaces gingival to the archwire were examined for the presence of white spot lesions of enamel,and a visual score based on the extent of demineralization were given. Result: Overall,76 % of subjects in the study who had white spot lesion were males and 24 % were females. In the 6- and 12-month groups,the percentages of individuals having visible white spot lesion were 37 % and 49 %, respectively. In the control group, only 9 % of the sample had white spot lesion. The 6-month and 12-month groups were significantly different from the control group but were not significantly different from each other.Conclusion: It is of great importance to evaluate the oral

  18. Disparities in herpes simplex virus type 2 infection between black and white men who have sex with men in Atlanta, GA.

    Okafor, Netochukwu; Rosenberg, Eli S; Luisi, Nicole; Sanchez, Travis; del Rio, Carlos; Sullivan, Patrick S; Kelley, Colleen F

    2015-09-01

    HIV disproportionately affects black men who have sex with men, and herpes simplex virus type 2 is known to increase acquisition of HIV. However, data on racial disparities in herpes simplex virus type 2 prevalence and risk factors are limited among men who have sex with men in the United States. InvolveMENt was a cohort study of black and white HIV-negative men who have sex with men in Atlanta, GA. Univariate and multivariate cross-sectional associations with herpes simplex virus type 2 seroprevalence were assessed among 455 HIV-negative men who have sex with men for demographic, behavioural and social determinant risk factors using logistic regression. Seroprevalence of herpes simplex virus type 2 was 23% (48/211) for black and 16% (38/244) for white men who have sex with men (p = 0.05). Education, poverty, drug/alcohol use, incarceration, circumcision, unprotected anal intercourse, and condom use were not associated with herpes simplex virus type 2. In multivariate analyses, black race for those ≤25 years, but not >25 years, and number of sexual partners were significantly associated. Young black men who have sex with men are disproportionately affected by herpes simplex virus type 2, which may contribute to disparities in HIV acquisition. An extensive assessment of risk factors did not explain this disparity in herpes simplex virus type 2 infection suggesting differences in susceptibility or partner characteristics.

  19. Liver spots

    Liver spots are changes in skin color that occur in older skin. The coloring may be due to aging, exposure to the sun or other sources of ultraviolet light, or causes that are not known. Liver spots are very common after age 40. They occur ...

  20. The 50-kDa protein of Apple chlorotic leaf spot virus interferes with intracellular and intercellular targeting and tubule-inducing activity of the 39-kDa protein of Grapevine berry inner necrosis virus.

    Isogai, M; Saitou, Y; Takahashi, N; Itabashi, T; Terada, M; Satoh, H; Yoshikawa, N

    2003-03-01

    To understand why transgenic Nicotiana occidentalis plants expressing a functional movement protein (MP) of Apple chlorotic leaf spot virus (ACLSV) show specific resistance to Grapevine berry inner necrosis virus (GINV), the MPs of ACLSV (50KP) and GINV (39KP) were fused to green, yellow, or cyan fluorescent proteins (GFP, YFP, or CFP). These fusion proteins were transiently expressed in leaf cells of both transgenic (50KP) and nontransgenic (NT) plants, and the intracellular and intercellular trafficking and tubule-inducing activity of these proteins were compared. The results indicate that in epidermal cells and protoplasts from 50KP plant leaves, the trafficking and tubule-inducing activities of GINV-39KP were specifically blocked while those of ACLSV-50KP and Apple stem grooving virus MP (36KP) were not affected. Additionally, when 39KP-YFP and 50KP-CFP were coexpressed in the leaf epidermis of NT plants, the fluorescence of both proteins was confined to single cells, indicating that 50KP-CFP interferes with the cell-to-cell trafficking of 39KP-YFP and vice versa. Mutational analyses of 50KP showed that the deletion mutants that retained the activities described above still blocked cell-to-cell trafficking of 39KP, but the dysfunctional 50KP mutants could no longer impede cell-to-cell movement of 39KP. Transgenic plants expressing the functional 50KP deletion mutants showed specific resistance against GINV. In contrast, transgenic plants expressing the dysfunctional 50KP mutants did not show any resistance to the virus. From these results, we conclude that the specific resistance of 50KP plants to GINV is due to the ability of the 50KP to block intracellular and intercellular trafficking of GINV 39KP.

  1. INFECTIOUS MYONECROSIS VIRUS (IMNV IN PACIFIC WHITE SHRIMP, Litopenaeus vannamei IN INDONESIA

    Taukhid Taukhid

    2008-12-01

    Full Text Available The aquaculture industry in Indonesia has been growing rapidly and plays an important role in rural development and export earning. Penaeid shrimp culture in Indonesia has become a leading export earning in fisheries sector. The main constraint encountered with shrimp culture has always been associated with disease outbreaks, especially, caused by viral agents. The Pacific white shrimp (Litopenaeus vannamei was unofficially introduced to Indonesia in 1999, and officially approved by Indonesian government in 2001. By the end of 2007, the Pacific white shrimp has been cultured in more than 17 provinces. The Taura Syndrome (TS disease was detected in Indonesia in 2002, and the disease is currently found in at least 10 provinces. The Infectious Myonecrosis (IMN is an emerging disease for L. vannamei in Indonesia, first detected in May-June 2006, causing significant mortalities in grow-out ponds. The IMN is characterized by an acute onset of gross signs: focal to extensive whitish necrotic areas in the striated muscle, especially on the distal abdominal segments and tail fan. White necrotic areas become reddened similar to the color of cooked shrimp. The outbreak resulted in elevated mortalities was initially associated with a chronic course of persistent low level mortalities. Up to date, IMN was detected in East Java, Bali, and West Nusa Tenggara provinces. This paper is a brief review of the epidemiological study of IMN disease of Pacific white shrimp in Indonesia: the status of outbreaks, surveillance, and disease diagnosis, and control measures.

  2. Peanut yellow spot virus is a member of a new serogroup of Tospovirus genus based on small (S) RNA sequence and organization.

    Satyanarayana, T; Gowda, S; Reddy, K L; Mitchell, S E; Dawson, W O; Reddy, D V

    1998-01-01

    Peanut yellow spot virus (PYSV) represents a distinct tospovirus species based on serology and nucleic acid hybridization. THe sequence of the S RNA was 2970 nucleotides with 22 nucleotide long inverted repeats (with three mismatches) at the termini. The coding was ambisense with a long open reading frame (ORF) in each strand. The 5'-large ORF (1,400 nucleotides in the viral sense (v) strand) encoded a protein with a predicted size of 53.2 kDa that was identified as the nonstructural (NSs) protein based on 16-21% sequence identity and 42-48% sequence similarity with other tospoviruses. A 3' ORF (741 nucleotides) in the virus complementary (vc) sense encoded a 28.0 kDa protein that was identified as the nucleocapsid (N) gene based on immuno-blot analysis of the in vitro expressed protein with PYSV polyclonal antiserum. The predicted N protein had 24-28% amino acid sequence identity and 44-51% sequence similarity with the members of other serogroups. In contrast to other tospoviruses, a third ORF (204 nucleotides) occurred in the vc strand, which could encode a protein with a predicted size of 7.5 kDa with two strong hydrophobic regions. The low degree of homology of N and NSs protein sequences with other serogroup members coupled with an additional ORF suggests that PYSV should be classified as a distinct species of the Tospovirus genus. This conclusion also is supported by the absence of serological cross reaction with other serogroups, and biological characteristics including thrips transmission, symptoms and host range.

  3. Effect of infiltration resin on the color masking of labial enamel white spot lesions%渗透树脂对前牙唇面白垩斑治疗效果的临床评价

    冯朝华; 刘荣芳; 赵奇; 楚小玉

    2013-01-01

    Objective To evaluate the effect of infiltration resin on masking white spot lesions by assessing the change in the white spot area. Methods Seventy-four maxillary anterior teeth with post-orthodontic decalcification teeth were investi-gated in this study. All teeth were treated with infiltration resin according to manufacturer recommendation. Standardized digital images were taken before, immediately after, and one week after treatment. The results were classified into three groups: com-pletely masked, partially masked and unchanged. The images of partially masked teeth were analyzed using an image analysis software. The size of the white spot lesion (W) and the whole-tooth facial surface (T) were measured, and the W︰T ratio(%) was calculated. Statistical evaluation of the lesions was performed using Kruskal-Wallis and Mann-Whitney tests. Results Among the 74 teeth, 20 (27%) teeth were classified as completely masked and 54 (73%) teeth were classified as partially masked; no tooth was unchanged. The W︰T ratio significantly decreased from 39.28% before treatment to 9.46% after treat-ment (P<0.05). Conclusion Resin infiltration is an effective treatment for masking white spot lesion. However, the masking effect depends on the lesion depth and activity.%目的: 评价渗透树脂对白垩斑颜色的改善效果。方法 选取正畸治疗后前牙白垩斑患牙74颗进行渗透树脂治疗。术前、术后即刻、术后1周拍摄照片。效果分为3级:有效、部分有效和无效。部分有效的患牙测量术前、术后白垩斑面积(W)及患牙唇面的面积(T),计算出W︰T比值(%)。采用Kruskal-Wallis and Mann-Whitney统计方法进行分析。结果 有效的患牙20颗(27%),部分有效的患牙54颗(73%),无效的患牙0颗。部分有效的患牙术前W︰T平均值是39.28%,术后W︰T平均值是9.46%(P<0.05)。结论 渗透树脂能明显改善白垩斑患牙的美观,改善程度与患牙脱矿程度有关。

  4. The Cucumber leaf spot virus p25 auxiliary replicase protein binds and modifies the endoplasmic reticulum via N-terminal transmembrane domains

    Ghoshal, Kankana [University of British Columbia, Faculty of Land and Food Systems, Vancouver, British Columbia, Canada V6T 1Z4 (Canada); Theilmann, Jane; Reade, Ron; Sanfacon, Helene [Agriculture and Agri-Food Canada Pacific Agri-Food Research Centre, 4200 Hwy 97, Summerland, British Columbia, Canada V0H 1Z0 (Canada); Rochon, D’Ann, E-mail: dann.rochon@agr.gc.ca [University of British Columbia, Faculty of Land and Food Systems, Vancouver, British Columbia, Canada V6T 1Z4 (Canada); Agriculture and Agri-Food Canada Pacific Agri-Food Research Centre, 4200 Hwy 97, Summerland, British Columbia, Canada V0H 1Z0 (Canada)

    2014-11-15

    Cucumber leaf spot virus (CLSV) is a member of the Aureusvirus genus, family Tombusviridae. The auxiliary replicase of Tombusvirids has been found to localize to endoplasmic reticulum (ER), peroxisomes or mitochondria; however, localization of the auxiliary replicase of aureusviruses has not been determined. We have found that the auxiliary replicase of CLSV (p25) fused to GFP colocalizes with ER and that three predicted transmembrane domains (TMDs) at the N-terminus of p25 are sufficient for targeting, although the second and third TMDs play the most prominent roles. Confocal analysis of CLSV infected 16C plants shows that the ER becomes modified including the formation of punctae at connections between ER tubules and in association with the nucleus. Ultrastructural analysis shows that the cytoplasm contains numerous vesicles which are also found between the perinuclear ER and nuclear membrane. It is proposed that these vesicles correspond to modified ER used as sites for CLSV replication. - Highlights: • The CLSV p25 auxiliary replicase targets the endoplasmic reticulum (ER). • Targeting of CLSV p25 is associated with ER restructuring. • Restructuring of the ER occurs during CLSV infection. • CLSV p25 contains 3 predicted transmembrane domains 2 of which are required for ER targeting. • Vesicles derived from the ER may be sites of CLSV replication.

  5. 西花蓟马传播番茄斑萎病毒研究进展%Reaearch advances on transmission of tomato spotted wilt virus by Frankliniella occidentalis (Pergande)

    杨帆; 刘万学; 张国; 万方浩

    2011-01-01

    Frankliniella occidentalis is one of the important quarantine pests in the world.This pest not only damages crops, but also transmits virus, thus causing serious economic loss to crops.As the F.occidentalis invaded into China, and the tomato spotted wilt virus it transmitted was found in various regions in China, it seriously threatened the cash crops due to its wide suitable distribution areas.This article summarizes the process that F.occidentalis gets, carries and transmits the tomato spotted wilt virus, and the cycle of the virus in F.occidentalis and the associated mechanism.This article also reviews the F.occidentalis - virus - plant interaction and its effect on the fitness of F.occidentalis, so as to provide a guide to the fundamental research and application in the transmission of tomato spotted wilt virus mediated by F.occidentalis.%西花蓟马Frankliniella occidentalis是世界性重要检疫性害虫之一,不仅直接取食危害作物而且传播病毒,从而造成极为严重的经济损失.由于西花蓟马在我国具有广泛的适生范围,随其入侵我国并随之传播的番茄斑萎病毒(Tomato spotted wilt virus)已在我国不同地域发现,对经济作物已形成严重威胁.本文综述了西花蓟马对番茄斑萎病毒的获取、携带和传播扩散过程及其病毒在蓟马体内的循环过程和机理,总结了影响西花蓟马传播番茄斑萎病毒效率的因素,并评述了"西花蓟马-病毒-植物"这一互作系统及其对西花蓟马生长发育适合度的影响,以期为我国西花蓟马传播番茄斑萎病毒的基础研究和防控提供理论依据与指导.

  6. Transcriptome analysis of Pacific white shrimp (Litopenaeus vannamei hepatopancreas in response to Taura syndrome Virus (TSV experimental infection.

    Digang Zeng

    Full Text Available BACKGROUND: The Pacific white shrimp, Litopenaeus vannamei, is a worldwide cultured crustacean species with important commercial value. Over the last two decades, Taura syndrome virus (TSV has seriously threatened the shrimp aquaculture industry in the Western Hemisphere. To better understand the interaction between shrimp immune and TSV, we performed a transcriptome analysis in the hepatopancreas of L. vannamei challenged with TSV, using the 454 pyrosequencing (Roche technology. METHODOLOGY/PRINCIPAL FINDINGS: We obtained 126919 and 102181 high-quality reads from TSV-infected and non-infected (control L. vannamei cDNA libraries, respectively. The overall de novo assembly of cDNA sequence data generated 15004 unigenes, with an average length of 507 bp. Based on BLASTX search (E-value <10-5 against NR, Swissprot, GO, COG and KEGG databases, 10425 unigenes (69.50% of all unigenes were annotated with gene descriptions, gene ontology terms, or metabolic pathways. In addition, we identified 770 microsatellites and designed 497 sets of primers. Comparative genomic analysis revealed that 1311 genes differentially expressed in the infected shrimp compared to the controls, including 559 up- and 752 down- regulated genes. Among the differentially expressed genes, several are involved in various animal immune functions, such as antiviral, antimicrobial, proteases, protease inhibitors, signal transduction, transcriptional control, cell death and cell adhesion. CONCLUSIONS/SIGNIFICANCE: This study provides valuable information on shrimp gene activities against TSV infection. Results can contribute to the in-depth study of candidate genes in shrimp immunity, and improves our current understanding of this host-virus interaction. In addition, the large amount of transcripts reported in this study provide a rich source for identification of novel genes in shrimp.

  7. SPOT Program

    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.

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

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

    2007-08-01

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

  9. Winter weeds as inoculum sources of tomato spotted wilt virus and as reservoirs for its vector, Frankliniella fusca (Thysanoptera: Thripidae) in farmscapes of Georgia.

    Srinivasan, Rajagopalbabu; Riley, David; Diffie, Stan; Shrestha, Anita; Culbreath, Albert

    2014-04-01

    Thrips-transmitted Tomato spotted wilt virus (TSWV) has a broad host range including crops and weeds. In Georgia, TSWV is known to consistently affect peanut, tomato, pepper, and tobacco production. These crops are grown from March through November. In the crop-free period, weeds are presumed to serve as a green bridge for thrips and TSWV. Previous studies have identified several winter weeds as TSWV and thrips hosts. However, their ability to influence TSWV transmission in crops is still not completely understood. To further understand these interactions, population dynamics of two prevalent vectors, viz., Frankliniella fusca (Hinds) and Frankliniella occidentalis (Pergande), on selected winter weeds were monitored from October through April in four counties from 2004 to 2008. Peak populations were typically recorded in March. F. fusca and F. occidentalis adults were found on winter weeds and their percentages ranged from 0 to 68% in comparison with other adults. Immatures outnumbered all adults. Microcosm experiments indicated that the selected winter weeds differentially supported F. fusca reproduction and development. The time required to complete one generation (adult to adult) ranged from 11 to 16 d. Adult recovery ranged from 0.97 to 2.2 per female released. In addition, transmission assays revealed that thrips efficiently transmitted TSWV from peanut to weeds, the incidence of infection ranged from 10 to 55%. Back transmission assays with thrips from TSWV-infected weeds resulted in up to 75% TSWV infection in peanut. These whole-plant transmission and back transmission assays provide the basis for TSWV persistence in farmscapes year round.

  10. 渗透树脂改善前牙正畸治疗后釉质白垩斑的初步研究%Clinical performance of infiltrated vestibular white spot lesions after orthodontic treatment

    李德利; 舒广; 陈立; 谭建国

    2012-01-01

    Objective To compare the efficacy of infiltration of vestibular white spot lesions (incipient caries) in incisors and canines shortly after removal of orthodontic brackets.Methods In this study,thirty subjects with age between 15 and 18 years old were selected and treated with one kind of infiltration resin:Icon-infiltrant (DMG,Hamburg,Germany).Standardized intraoral images were taken before and after treatment for 1 month,3 months and 6 months.Image analysis software:Image-Pro Plus 6.0 (Media Cybernetics,U S A) was used to quantify the visible areas of the demineralized lesions.The measurement data before and after using the infiltrant resin for 1 month,3 months and 6months were analyzed using SPSS 20.0 software package.Results In the incisor,the aera of white spot lesions was greatly reduced after using it[(29.29 ± 9.62)%] and for 1 month[(22.21 ±6.94)%],3 months [(26.43 ± 7.85) %] and 6 months [(22.82 ± 8.11) %] compare to that before treatment [(41.36 ± 13.21)%] (P< 0.001).And the outcome of the lateral incisor and the canine was similar to the incisor.Conclusions During the observing period of this study,infiltrant resin is effective in reducing the white spot lesion size,but further researches are needed for long-term results.%目的 评价树脂渗透技术改善前牙正畸治疗后脱矿形成的釉质白垩斑的美学修复效果及其稳定性.方法 选取已完成正畸治疗的前牙区釉质白垩斑患者30例,采用自身配对实验设计,使用渗透树脂(Icon-Infiltrant(R))对白垩斑病损进行治疗.通过拍摄标准化数码照片,使用Image Pro Plus6.0图像分析软件记录渗透树脂治疗前,治疗后即刻、1个月、3个月及6个月时白垩斑面积的变化.结果 各牙位釉质白斑面积占牙冠部唇面面积百分比的变化表现相同,中切牙治疗前[(41.36 ±13.21)%]与治疗后即刻[(29.29±9.62)%]、治疗后1个月[(22.21±6.94)%]、3个月[(26.43±7.85)%]及6个月[(22.82±8.11)

  11. Optimization for Prokaryotic Expression of MCP Gene of Red-spotted Grouper Nervous Necrosis Virus%赤点石斑鱼神经坏死病毒MCP基因原核表达条件优化

    苏友禄; 冯娟; 孙秀秀; 郭志勋; 闫云锋; 黄剑南

    2008-01-01

    [Objective] To optimize the prokaryotic expression of MCP gene of red-spotted grouper nervous necrosis virus. [Method] The MCP gene was amplified from red-spotted grouper nervous necrosis viral genome by RT-PCR. The recombinant expression vector pRSET A-MCP was constructed and transformed into BL21(DE3)plysS to express proteins with induction in different media, at different pH, or at different temperatures. [Result] The expression level of recombinant bacteria reached a peak with induction under the following condition: SOB or LB medium, pH 7.0, 37 ℃ while the fusion protein was about 44.5 kD in molecular weight. [Conclusion] This study provided a basis for the development of RGNNV-MCP vaccine.

  12. Virus-phytoplankton adhesion: a new WSSV transmission route to zooplankton

    2007-01-01

    The pathogenicity of white spot syndrome virus (WSSV) to zooplankton species, rotifer Brachionus urceus (Linnaeus), copepod Acartia clausi (Giesbrecht) and mysid shrimp Neomysis awatschensis (Brandt), was estimated by immersion challenge and virus-phytoplankton adhesion route to investigate a potential new transmission route of WSSV to zooplankton. WSSV succeeded in infecting these zooplankton species and nested-PCR revealed positive results for the virus-phytoplankton adhesion route, whereas WSSV cannot infect zooplankton by immersion challenge. These results indicated that virus-phytoplankton adhesion route is a successful new transmission route of WSSV to zooplankton and also implied that phytoplankton could be a carrier in WSSV transmission.

  13. 渗透树脂用于正畸后白垩斑治疗的研究进展%Research of infiltration resin for the treatment of post-orthodontic white spot lesions

    李昕彧(综述); 沈兰花; 邵帅(审校)

    2016-01-01

    釉质脱矿在正畸固定矫治器的正畸期间发病率较高,形成白垩斑,影响牙齿美观。渗透树脂以其微创、无痛的特点使患者易于接受。它可阻断病变继续发展,并且可改善白垩斑外观。本文就渗透树脂颜色稳定性、显微硬度等特点进行阐述。%Enamel demineralization usually occurs during orthodontic treatment which is found white spots that af⁃fect the esthetic of teeth. Characterized by minimally invasive and painless, infiltration resin can be easily accepted by the patients which blocks the development of caries and improves the appearance of the teeth. This paper discusses the characteristics of infiltration resin such as color stability and micro⁃hardness.

  14. Efficacy of one year treatment of icon infiltration resin on post-orthodontic white spots%渗透树脂治疗正畸后牙面白垩斑的一年疗效观察

    冯朝华; 楚小玉

    2013-01-01

    To clinically evaluate the effectiveness of icon infiltration resin on masking post-orthodontic white spots. Methods; Eight post-orthodontic patients with 6 maxillary anterior teeth showing signs of decalcification (total 48 teeth) were enrolled in this study. All teeth were treated with icon resin infiltration according to manufacturer' s recommendation. Standardized digital photographs were taken before, immediately after and 1 week, 6 and 12 months after treatment. Before taking pictures the assigned teeth were cleaned using pumice and rubber polishing cups. The results were classified into three groups: completely masked, partially masked, and unchanged. Pictures of partially masked teeth were analyzed using image analysis software (Image-pro plus 6.0) , size of the white spot lesion (W) and the whole tooth facial surface (T) were measured, then W/T ratio (in % ) was calculated. The images were imported into image analysis software ( Photoshop) which presented the images into histograms of gray scale from (0 to 255). Results; Among the 48 teeth, 11 teeth (22. 9% , 11/48) were classified as completely masked, whereas 37 teeth (11. I % , 37/48) were classified as partially masked and no tooth unchanged. For partially masked teeth, W/T ratio decreased significantly after treatment from 31. 37% to 7.99% (by Wilcoxon's signed rank test, P 0. 05) in comparison to 1 week after treatment. Conclusion: Resin infiltration was proven to be an effective treatment for masking white spot lesions. The surface color of infiltrated lesions remained stable after 12 months.%目的:评价渗透树脂对正畸后牙面脱矿(白垩斑)的治疗效果及颜色稳定性.方法:对正畸治疗后6颗上颌前牙均出现脱矿的患者8名(患牙共48颗)进行渗透树脂治疗.术前、术后即刻、术后1周、6个月和12个月分别在同等条件下拍摄照片.效果分为3级:有效、部分有效和无效.部分有效的患牙测量术前、术后白垩斑(white spot,W)及

  15. Efficacy of CMX001 as a Prophylactic and Presymptomatic Antiviral Agent in New Zealand White Rabbits Infected with Rabbitpox Virus, a Model for Orthopoxvirus Infections of Humans

    Richard W. Moyer

    2011-01-01

    Full Text Available CMX001, a lipophilic nucleotide analog formed by covalently linking 3‑(hexdecyloxypropan-1-ol to cidofovir (CDV, is being developed as a treatment for smallpox. CMX001 has dramatically increased potency versus CDV against all dsDNA viruses and, in contrast to CDV, is orally available and has shown no evidence of nephrotoxicity in healthy volunteers or severely ill transplant patients to date. Although smallpox has been eliminated from the environment, treatments are urgently being sought due to the risk of smallpox being used as a bioterrorism agent and for monkeypox virus, a zoonotic disease of Africa, and adverse reactions to smallpox virus vaccinations. In the absence of human cases of smallpox, new treatments must be tested for efficacy in animal models. Here we first review and discuss the rabbitpox virus (RPV infection of New Zealand White rabbits as a model for smallpox to test the efficacy of CMX001 as a prophylactic and early disease antiviral. Our results should also be applicable to monkeypox virus infections and for treatment of adverse reactions to smallpox vaccination.

  16. MONEY LAUNDERING AS AN ISOLATED PHENOMENON AND LEADING FACTOR IN WHITE-COLLAR CRIMINALITY, PUTS MOLDOVA CASE JUDICIARY ON THE SPOT

    Jacob RUB

    2015-11-01

    Full Text Available Money laundering is a leading factor in white-collar criminality, with a big effect upon Gross National Product of the Israeli and Moldova economies. Israel and Moldova have made the first steps In order to cope with money laundering phenomenon. The model of handling the phenomenon in the USA has been studied that constitutes a role model of fighting money laundering. The vast majority (77% of all frauds were committed by individuals working in one of six departments: accounting, operations, sales, executive/upper management, customer service and purchasing. In Moldova, money laundering for the most part is connected to traditional forms and activities sources of illegal proceeds .Moldova is a transit country for money laundering. A money-laundering scandal is casting Moldova’s judiciary in an unfavorable light and is raising concerns about the government’s commitment to reforms needed to keep European Union integration on track. It is a need to address corruption as a national priority, and the priority of the relations between the EU and Moldova. The money laundering scandal indicates that Moldova is reluctant to wade deeply into judicial reform. In fact, the laundered of $20 billion, is an amount more than twice the size of Moldova’s GDP in 2013, and may be that it is just the tip of the iceberg, probably. White-collar criminality in Israel and Moldova means success of coping of the enforcement and judicial systems. In order to cope with this phenomenon, Israel makes its first steps. Anyway, the message must pave the way through a creation of a new model for dealing with reduction the isolated phenomenon of money laundering in both states.SPĂLAREA DE BANI, UN FENOMEN IZOLAT ŞI UN FACTOR IMPORTANT ÎN CRIMINALITATEA GULERELOR ALBE, PUNE PRACTICA JUDICIARĂ DIN REPUBLICA MOLDOVA INTR-O SITUAŢIE DIFICILĂSpălarea de bani este un factor important în criminalitatea gulerelor albe, având un efect semnificativ asupra produ-sului na

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

    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.

  18. Occurrence and Distribution of Papaya ring spot virus and Papaya leaf-distortion mosaic virus in Hainan Province%海南番木瓜PRSV和PLDMV病毒发生情况及分子鉴定

    张雨良; 黄启星; 郭安平; 曾会才; 赵辉; 孔华; 刘志昕

    2013-01-01

    对海南昌江、乐东、澄迈、文昌和三亚5个主要番木瓜产区的病毒病发生情况进行调查.对76份番木瓜疑似病毒样品进行检测,检测结果表明海南番木瓜病毒病主要有2种:由番木瓜环斑病毒(Papaya ring spot virus,PRSV)引起的番木瓜环斑病毒病和由番木瓜畸形花叶病毒(Papaya leaf-distortion mosaic virus,PLDMV)引起的番木瓜畸形花叶病毒病.其中,番木瓜环斑病毒病最为常见,检出率达77.63%,是影响海南地区番木瓜产业健康发展的主要病毒病:畸形花叶病毒病发生率很低,仅检出2例番木瓜畸形花叶病毒病样品.以PRSV病毒基因组P1和Hc-Pro作为靶标基因进行分子克隆、测序,结果表明海南地区PRSV病毒可明显分为2个株系:HNPRSV-1株系和HN-PRSV-2株系.此外,基于CP基因的系统进化树分析结果表明在海南新发现的PLDMV病毒可能源于台湾和日本.本研究可为开展番木瓜花叶病的防控和创制抗花叶病毒番木瓜新种质提供数据参考.

  19. Mongolian spots

    Divya Gupta

    2013-01-01

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

  20. 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

    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, ab

  1. 凡纳滨对虾感染与暴发白斑综合征(WSD)的密度因素分析%Investigation of the Effect of Aquaculture Density on the Infection and Outbreak of White Spot Disease (WSD) in Litopenaeus vannamei

    周俊芳; 杨先乐; 万夕和; 胡琳琳; 周帅; 房文红

    2011-01-01

    White spot syndrome viruse (WSSV) carried by cultured Litopenaeus vannamei in several districts in Shanghai, Jiangsu and Hainan in China was detected by polymerase chain reaction (PCR). Data showed that up to 40%-58% of the sampled shrimp were infected with WSSV, indicating that the infection rate of WSSV in L. vannamei was still high in the latest years even if the shrimp were of SPF or SPR origin. As a possible risk factor associated with the consequence, the current aquaculture model characterized by high culture density, was considered in this study, and the roles of high and low culture density during shrimp infection with WSSV and outbreak of WSD were investigated, respectively. Analysis of the shrimp mortality indicated that the mortality of shrimp cultured in the high density was 96.9% (with salinity unchanged) or 98.1% (with abrupt salinity reduction), whereas that of those cultured in the low density was 5.0 % (with salinity unchanged) or 26.7 % ( with abrupt salinity reduction), suggesting that the difference in mortality between shrimp cultured with high and low density was remarkable;data from real time PCR also indicated that WSSV multiplicated much more quickly in shrimp cultured in the high density than that in the low density. Taken together, the present study demonstrates that the high culture density plays an important role during shrimp infection with WSSV and outbreak of WSD in current intensive aquaculture systems.%采用PCR检测技术分别对上海、江苏、海南部分养殖地区凡纳滨对虾携带重要病毒性病原--白斑综合征病毒(WSSV)的情况进行了调查分析.结果显示40%~58%的受检测对虾携带WSSV,表明近年在养殖SPF(无特定病原)甚至SPR(抗特定病原)亲虾子代的情况下,凡纳滨对虾的WSSV感染率仍然很高.为探讨现行集约化、高密度养殖模式对凡纳滨对虾感染与暴发白斑综合征(white spot disease,WSD)的影响,设高、低密度2个养殖模拟

  2. Esthetic effect of resin infiltration for the treatment of post-orthodontic white spot lesions%渗透树脂治疗正畸后白垩斑美学效果的临床研究

    谷希; 高源; 杨琳; 张君平; 李继遥

    2015-01-01

    目的:分析渗透树脂治疗正畸后白垩斑的美学效果并与微研磨法进行比较。方法:筛选符合纳入排除标准的正畸后白垩斑患者10名(66颗患牙),采用自身对照设计,将不同区域患牙随机分为渗透树脂组和微研磨组(n =33)。处理后采用口内照相和图像软件分析治疗前(T0)、1周复诊(T2)、3月复诊(T3)时患牙的病损面积及唇面总面积,计算出病损面积比(R值)及治疗有效率(SR 值),比较2种方法治疗正畸后白垩斑的牙齿美学恢复情况。采用重复测量单因素方差分析对结果进行统计。结果:渗透树脂可以显著减患牙小白垩斑病损面积(P <0.05),且3个月内保持稳定;渗透树脂组和微研磨组治疗有效率分别为88.68%和64.67%(P <0.05)。结论:渗透树脂改善正畸后白垩斑病损的美观效果优于微研磨法。%Objective:To evaluate the effects of resin infiltration in the treatment of post-orthodontic white spot lesions (WSLs)and compare it with microabrasion.Methods:1 0 subjects with 66 teeth with noncavitated and unrestored WSLs after multibracket treatment were recruited.A simple randomized,split-mouth and controlled design was used to allocate WSLs to the resin infiltration and micro-abrasion groups(n =33).The area of the WSLs were measured photographically by using image analysis software before treatment (T0),1 week(T2)and 3 months(T3)after treatment.Data were analyzed with multifactorial analysis of the variances with repeated measures at an a level of 5% and a power of 80%.Results:The area of the white spot lesions(R)decreased significantly in both groups(P <0.05).At T3 the success rate of resin infiltration group(88.68%)was significantly higher than that of microabrasion group (64.67%)(P <0.05).Conclusion:Resin infiltration is more effective than microabrasion in the treatment of post-orthodontic WSLs.

  3. Intracellular distribution, cell-to-cell trafficking and tubule-inducing activity of the 50 kDa movement protein of Apple chlorotic leaf spot virus fused to green fluorescent protein.

    Satoh, H; Matsuda, H; Kawamura, T; Isogai, M; Yoshikawa, N; Takahashi, T

    2000-08-01

    The 50 kDa protein (50KP) encoded by ORF2 of Apple chlorotic leaf spot virus (ACLSV) fused to green fluorescent protein (GFP) was expressed transiently in cells of Nicotiana occidentalis and Chenopodium quinoa leaves. Its intracellular distribution, cell-to-cell trafficking in leaf epidermis and tubule formation on the surface of protoplasts were analysed. The 50KP-GFP fluorescence was distributed as small irregular spots or a fibrous network structure on the periphery of epidermal cells and protoplasts of both plant species. In leaf epidermis of N. occidentalis, the protein spread from the cells that produced it into neighbouring cells in both young and mature leaves and targetted plasmodesmata in these cells. In contrast, GFP was restricted to single cells in most cases in mature leaves. When 50KP and GFP were co-expressed in leaf epidermis of N. occidentalis, GFP spread more widely from the initial cells that produced it than when GFP was expressed alone, suggesting that 50KP facilitated the cell-to-cell trafficking of GFP. 50KP-GFP was able to complement local spread of 50KP-deficient virus when expressed transiently in leaf epidermis of C. quinoa. Expression of 50KP-GFP in protoplasts resulted in the production of tubular structures protruding from the surface. Mutational analyses showed that the C-terminal region (aa 287-457) was not essential for localization to plasmodesmata, cell-to-cell trafficking, complementation of movement of 50KP-deficient virus or tubule formation on protoplasts. In contrast, deletions in the N-terminal region resulted in the complete disruption of all these activities.

  4. Pathogenesis of spring viremia of carp virus in emerald shiner Notropis atherinoides Rafinesque, fathead minnow Pimephales promelas Rafinesque and white sucker Catostomus commersonii (Lacepede).

    Misk, E; Garver, K; Nagy, E; Isaac, S; Tubbs, L; Huber, P; Al-Hussinee, L; Lumsden, J S

    2016-06-01

    Spring viremia of carp (SVC) is a reportable disease to the World Organization of Animal Health (OIE) as it is known to cause significant international economic impact. In Canada, the first and only isolation of SVC virus (SVCV) was in 2006, from common carp Cyprinus carpio L., at Hamilton Harbour, Lake Ontario. The susceptibility of fathead minnow Pimephales promelas Rafinesque, emerald shiner Notropis atherinoides Rafinesque and white sucker Catostomus commersonii (Lacepede) to intraperitoneal injection of the Canadian isolate (HHOcarp06) was evaluated using experimental infection, virus isolation, quantitative reverse transcription polymerase chain reaction (qRT-PCR), histopathology and immunohistochemistry (IHC). Emerald shiner and fathead minnow were most susceptible with 43 and 53% cumulative mortality, respectively, compared with koi at 33%. Quantitative RT-PCR demonstrated that koi had high viral loads throughout the experiment. At 34 days post-infection, SVCV was detected from sampled emerald shiner and white sucker in very low titre and was not detected from fathead minnow. Koi, fathead minnow and emerald shiner had gross lesions typical of SVC disease. The histopathological picture was mostly dominated by necrotic changes in kidney, spleen, liver, pancreas and intestine. IHC further confirmed SVCV infection, and staining was largely correlated with histological lesions.

  5. Rocky Mountain Spotted Fever

    ... facebook share with twitter share with linkedin Rocky Mountain Spotted Fever Rocky Mountain spotted fever is a ... New Mexico. Why Is the Study of Rocky Mountain Spotted Fever a Priority for NIAID? Tickborne diseases ...

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

    Witteveldt, J.; Vlak, J.M.; Hulten, van 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 Pen

  7. White and red blood cell picture in rabbits experimentally infected with strains of the rabbit haemorrhagic disease (RHD) virus without or with variable haemagglutination capacity.

    Niedźwiedzka-Rystwej, P; Tokarz-Deptuła, B; Deptuła, W

    2016-12-01

    The aim of the study was to establish if haemagglutination of rabbit haemorrhagic disease virus (RHDV) affects haematological picture of peripheral blood in rabbits and the pathogenicity of the virus. The study analyzed white and red blood cell picture in rabbits experimentally infected with two non-haemagglutinating (HA-) RHDV strains (Frankfurt and Asturias) and one strain with variable haemagglutination capacity (HA+/-) (Hagenow). Studies with HA- and HA +/- are rare and relate only to 4 HA- strains (2 RHDV: BLA and Rainham; 2 RHDVa: Pv97 and 9905) and 1 HA+/- RHDV strain: ŻD, where less changes in haematological indices and less pathogenicity were observed. We found that changes caused by HA- Frankfurt strain were related to the number of neutrophils and thrombocytes, while in HA- strain Asturias, in thrombocytes and leukocytes. Changes evoked by HA+/- Hagenow strain pertained to the number of eosinophils, thrombocytes, leukocytes, monocytes, and concentration of hemoglobin. Mortality caused by the Frankfurt strain was 100% between 36 and 48 h post infection (p.i.), while that caused by Asturias strain was 100% between 24 and 36 h p.i., and that observed in case of Hagenow strain was 90% between 36 and 48 h p.i. The changes in haematological picture caused by the HA- and HA+/- RHDV strains were less intensive than those found in case of the HA+ RHDV strains, which cannot be confirmed for pathogenicity, and is not in line with the existing hypothesis suggesting higher pathogenicity in HA+ viruses.

  8. Are 'hot spots' hot spots?

    Foulger, Gillian R.

    2012-07-01

    The term 'hot spot' emerged in the 1960s from speculations that Hawaii might have its origins in an unusually hot source region in the mantle. It subsequently became widely used to refer to volcanic regions considered to be anomalous in the then-new plate tectonic paradigm. It carried with it the implication that volcanism (a) is emplaced by a single, spatially restricted, mongenetic melt-delivery system, assumed to be a mantle plume, and (b) that the source is unusually hot. This model has tended to be assumed a priori to be correct. Nevertheless, there are many geological ways of testing it, and a great deal of work has recently been done to do so. Two fundamental problems challenge this work. First is the difficulty of deciding a 'normal' mantle temperature against which to compare estimates. This is usually taken to be the source temperature of mid-ocean ridge basalts (MORBs). However, Earth's surface conduction layer is ˜200 km thick, and such a norm is not appropriate if the lavas under investigation formed deeper than the 40-50 km source depth of MORB. Second, methods for estimating temperature suffer from ambiguity of interpretation with composition and partial melt, controversy regarding how they should be applied, lack of repeatability between studies using the same data, and insufficient precision to detect the 200-300 °C temperature variations postulated. Available methods include multiple seismological and petrological approaches, modelling bathymetry and topography, and measuring heat flow. Investigations have been carried out in many areas postulated to represent either (hot) plume heads or (hotter) tails. These include sections of the mid-ocean spreading ridge postulated to include ridge-centred plumes, the North Atlantic Igneous Province, Iceland, Hawaii, oceanic plateaus, and high-standing continental areas such as the Hoggar swell. Most volcanic regions that may reasonably be considered anomalous in the simple plate-tectonic paradigm have been

  9. Investigation into the incidence and severity of white spot lesions after fixed orthodontic treatment through Medical Image J software%医学图像处理软件Image J在评估正畸固定矫治后牙釉质脱矿白斑程度中的作用

    于海涛; 古力巴哈·买买提力; 陈文君

    2014-01-01

    Objective The purpose of this study was to investigate the incidence and the mean coverage rate of white spot lesions through medical Image J software and to evaluate the severity of enamel demineraliza-tion exactly.Methods 208 clinical digital photos of patients who received fixed orthodontic treatment and the gingival index of patients examined in oral were collected.Then the incidence and the mean coverage rate of white spot lesions of the labial surface of 8 maxillary anterior teeth were evaluated through medical Image J software.Based on the data,this paper analyzed and compared whether the white spot lesions would appear on the particular tooth positions and whether the area of the white spot lesions would be lar-ger on the particular tooth positions.Whether the difference of the incidence and the mean coverage rate of white spot lesions between different genders exist or not would be examined.Comprehensive evaluation of relationship would be carried out between the incidence and the mean coverage rate of white spot lesions and the gingival index of patients.Results The incidence of at least 1 white spot lesion on the labial sur-face of the maxillary anterior teeth was 31.3% and the average white spot lesion coverage was 8.4%.The order of incidence was lateral incisor(34.8%),canine (28.8%),premolar (26.9%),and central incisor (20.7%).It was no difference between sex .The higher the gingival index of patients was,the higher the grade of white spot lesion was.Conclusion The incidence and severity of white spot lesions on the maxil-lary anterior teeth could be evaluated effectively through medical Image J software.%目的:探讨医学图像处理软件 Image J在评估正畸固定矫治后牙釉质脱矿白斑的发生率、平均覆盖率及严重程度中的作用。方法收集208例患者完成固定正畸矫治后的临床数码照片和口内检查患者的牙龈指数,应用医学图像处理软件 Image J评估8颗上颌前牙颊面的牙釉质脱

  10. The efficacy of varnish XT and ICON resin infiltration treatment on the color of white spot lesions%Varnish XT和ICON渗透树脂对釉质龋白斑颜色的影响

    郭静; 姚悦; 包旭英; 赵晓一

    2015-01-01

    目的:研究Varnish XT和ICON渗透树脂两种微创治疗材料对釉质龋白斑颜色的影响。方法:选择牛门齿100颗,将唇面向下作为观察面,环氧树脂包埋,打磨。暴露至少6mm×10mm的釉质牙面,随机分A,B,C,D,E 5个区,E区用抗酸指甲油封闭。并分别采用人工龋脱矿液后A区不做处理,B区经Varnish XT处理,C区经ICON渗透树脂处理,D区经氟化物处理后,用电子分光光度比色仪测量L*值。结果:脱矿后样本A、 B、 C、 D较E区升高, L*值有统计学差异(P0.05),尚不能认为两区的L*值不同。%Objective:To study the effect of two minimally invasive surgical treatment materials, Varnish XT and ICON penetrating resin on the color alteration of enamel white spots.Methods:100 incisors of cow were selected, leaving lip side down as the observation side, embedded with ethoxyline resin, and polished. Exposed area of each specimen was at least 6mm×10mm on the enamel face, and divided into five regions (A-E). Region E was sealed with antiacid nail polish before demineralization. The artificial caries was realized using a demineralization solution., Each region was treated with Varnish XT for region B, ICON resin infiltration for region C, and fluoride for region D, respectively, while Region A received no treatment. Electronic spectrophotometer was used to measure L*value, and micro hardness test of the surface micro-hardness of each region was analyzed.Results:After demineralization L* value of A, B, C, D increased remarkably. After treatment, each L*value of Region B, C and D was significantly lower than that of region A (P0.05).

  11. Planning for Rift Valley fever virus: use of geographical information systems to estimate the human health threat of white-tailed deer (Odocoileus virginianus-related transmission

    Sravan Kakani

    2010-11-01

    Full Text Available Rift Valley fever (RVF virus is a mosquito-borne phlebovirus of the Bunyaviridae family that causes frequent outbreaks of severe animal and human disease in sub-Saharan Africa, Egypt and the Arabian Peninsula. Based on its many known competent vectors, its potential for transmission via aerosolization, and its progressive spread from East Africa to neighbouring regions, RVF is considered a high-priority, emerging health threat for humans, livestock and wildlife in all parts of the world. Introduction of West Nile virus to North America has shown the potential for “exotic” viral pathogens to become embedded in local ecological systems. While RVF is known to infect and amplify within domestic livestock, such as taurine cattle, sheep and goats, if RVF virus is accidentally or intentionally introduced into North America, an important unknown factor will be the role of local wildlife in the maintenance or propagation of virus transmission. We examined the potential impact of RVF transmission via white-tailed deer (Odocoileus virginianus in a typical north-eastern United States urban-suburban landscape, where livestock are rare but where these potentially susceptible, ungulate wildlife are highly abundant. Model results, based on overlap of mosquito, human and projected deer densities, indicate that a significant proportion (497/1186 km2, i.e. 42% of the urban and peri-urban landscape could be affected by RVF transmission during the late summer months. Deer population losses, either by intervention for herd reduction or by RVF-related mortality, would substantially reduce these likely transmission zones to 53.1 km2, i.e. by 89%.

  12. The efficacy of ICON resin infiltration treatment on the color and microhardness of white spot lesions%ICON渗透树脂对釉质龋白斑颜色及表面微硬度的影响

    郭静; 周瑾; 黄晓琳; 吴佩玲

    2015-01-01

    Objective To study the effect of two minimally invasive surgical treatment materials including Varnish XT and ICON resin infiltration,on the color and surface microhardness of enamel caries white spot.Methods 100 incisors of cow were selected,leaving lip side down as the observation side,ethoxyline resin was embedded and polished.Exposed an area of at least 6 mm×10 mm on the enamel face,and divid-ed into A,B,C,D four regions,after treated with demineralization liquid for artificial caries,B region was adopted no treatment for,C region was treated with fluoride D region was treated with ICON resin in-filtration,and antiacid nail polish sealing was used before demineralization.Electronic spectrophotometer was used to measure L*value,and microhardness meter test of the surface micro-hardness of each re-gions was also undertaken.Results After demineralization,L* value of B,C,D of all samples increased remarkably;After treatment,L*values of C,D of all samples decreased compared with B region,and it was statistical significance (P 0.05).Based on test of the surface micro-hardness of each regions,after treatment,SMH of B,C,D of all samples decreased remarkably.SMH of C,D of all samples increased compared with B re-gion,Meanwhile Bwhite spot of caries and has obvious improvement effect on SMH,ICON resin infiltration is superior to fluoride.%目的:研究ICON渗透树脂微创治疗材料对釉质龋白斑颜色和表面微硬度的影响。方法选择牛门齿100颗,将唇面向下作为观察面,环氧树脂包埋、打磨。暴露至少6 mm×10 mm的釉质牙面,分 A、B、C、D 4个区,A区用抗酸指甲油封闭。采用人工龋脱矿液脱矿后,B区不做处理,C区经氟化物处理,D区经 ICON渗透树脂处理,用电子分光光度比色仪测量 L*值,用维氏硬度仪测量表面显微硬度值。结

  13. Varnish XT和ICON渗透树脂对釉质龋白斑微硬度的影响%Effect of Varnish XT and ICON resin infiltration treatment on the microhardness of white spot lesions

    郭静; 赵晓一; 吴佩玲

    2014-01-01

    背景:早期龋的治疗方法中,可以采用氟化物对龋白斑进行再矿化处理。长效释氟玻璃离子保护膜(Varnish XT)和ICON渗透树脂是两种新型的材料,Varnish XT是一种新型的树脂加强型玻璃离子,可以作为再矿化材料的新选择。ICON渗透树脂是一种流动性较好的高渗透系数树脂,借助毛细作用渗入到脱矿釉质的微孔中,利用低黏性的渗透树脂取代由于脱矿而丧失的硬组织并占据微孔的空间,填补釉质脱矿区并阻止病损的进一步发展。目的:观察Varnish XT和ICON渗透树脂两种微创治疗材料对釉质龋白斑微硬度的影响。方法:选择牛门齿100颗,将唇面向下作为观察面,环氧树脂包埋,打磨。暴露至少6 mm×10 mm的釉质牙面,由切端至颈部依次分A,B,C,D,E 5个区,E区用抗酸指甲油封闭。分别采用人工龋脱矿液后,A区不做处理,B区经Varnish XT处理,C区经ICON渗透树脂处理,D区经氟化物处理后,用韦氏硬度仪测量表面显微硬度值。结果与结论:脱矿样本经处理后A,B,C,D区的表面显微硬度值较E区均明显降低(P B区、D区(P 0.05)。说明ICON渗透树脂、Varnish XT和氟化物对龋白斑的微硬度都有明显的改善,ICON渗透树脂的改善较Varnish XT和氟化物明显,Varnish XT和氟化物之间无明显差异。%BACKGROUND:In the treatment of early caries, fluoride can be used for remineralization of white spot lesions. Varnish XT (durable fluoride-releasing coating) and ICON penetration resins are two new materials. Varnish XT as a new type of resin reinforced glass ionomer can be selected as mineralized material. ICON penetration resin is a high-permeability resin with good liquidity that can infiltrate by capilary action into the pores created by enamel demineralization. Low-viscosity resin is used to replace the lost hard tissue due to demineralization and occupy the micropores, so as

  14. 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

    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.

  15. 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

    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

  16. Rocky Mountain spotted fever

    ... Mountain spotted fever is caused by the bacteria Rickettsia rickettsii (R. Rickettsii) , which is carried by ticks. ... Saunders; 2014:chap 212. Walker DH, Blaton LS. Rickettsia rickettsii and other spotted fever group rickettsiae (Rocky ...

  17. [Genetic characterization of viruses from the antigenic complex Tyuleniy (Flaviviridae, Flavivirus): Tyuleniy virus (TYUV) (ID GenBank KF815939) isolated from ectoparasites of colonial seabirds--Ixodes (Ceratixodes) uriae White, 1852, ticks collected in the high latitudes of Northern Eurasia--and Kama virus (KAMV) isolated from the Ixodes lividus Roch, 1844, collected in the digging colonies of the middle part of Russian plane].

    L'vov, D K; Al'khovskiĭ, S V; Shchelkanov, M Iu; Shchetinin, A M; Deriabin, P G; Samokhvalov, E I; Gitel'man, A K; Botikov, A G

    2014-01-01

    Genetic research into the Tyuleniy virus (TYUV) (ID GenBank KF815939) isolated in high latitudes from the Ixodes uriae White, 1852, ticks collected in the nesting colonies of the Alcidae (Leach, 1820) birds and Kama virus (KAMV) (ID GenBank KF815940) isolated from the I. lividus ticks collected in the nesting bird colonies in the middle part of the Russian Plane was carried out. Full-genome comparative analysis revealed 70% homology between KAMV and TYUV on the nucleotide level and 74% on the amino acid level. Thus, KAMV is a new member of the TYUV complex belonging to the seabird tick-borne virus group (STBVG) of Flavivirus (Flaviviridae). KAMV is a separate virus and forms separate phylogenetic line together with the TYUV, Meaban virus (MEAV), and Saumarez Reef virus (SREV).

  18. Aangifteplichtige schaaldierziekten, deel 2: White Spot Syndrome of White Spot Disease

    Haenen, O.L.M.; Engelsma, M.Y.

    2008-01-01

    De nieuwe aquacultuurrichtlijn 2006/88/EG, die in de herfst van 2006 is goedgekeurd door het Europese Parlement bevat een lijst van aangifteplichtige vis-, schaal en schelpdierziekten. In deze aflevering gaan we in op een virale ziekte van schaaldieren die op delijst van niet-exotische ziekten staat

  19. Expression and Purification of Major Capsid Protein of Orange-spotted Grouper Nervous Necrosis Virus%斜带石斑鱼神经坏死病毒主衣壳蛋白的原核表达与纯化

    陈晓艳; 翁少萍; 殷志新; 何建国

    2005-01-01

    将含有斜带石斑鱼Epinephelus coioides神经坏死病毒(orange-spotted grouper nervous necrosis virus,OGNNV)主衣壳蛋白(major capsid protein, MCP)基因的重组表达质粒载体pET32a-MCP转化到大肠杆菌BL21(DE3)中进行融合表达,经SDS-PAGE分析和Western-blot鉴定,证实了重组大肠杆菌融合表达了斜带石斑鱼神经坏死病毒主衣壳蛋白.表达的融合蛋白主要以不可溶的包涵体形式存在,提取的包涵体中融合蛋白含量占60%以上,经柱层析纯化蛋白,纯化度达90%以上.

  20. 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)

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

    2008-01-01

    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, Hi...

  1. Transferring cucumber mosaic virus-white leaf strain coat protein gene into Cucumis melo L. and evaluating transgenic plants for protection against infections

    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.

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

    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/μg of total RNA in the hemolymph, 5.096×10(5) to 1.26×10(3)copies/μg in the pleopods, 6.85×10(8) to 3.09×10(4)copies/μg in muscle and 8.15×10(6) to 3.90×10(3)copies/μg in gills. Different viral loads of IMNV were found with greater values in the hemolymph and muscle, followed by the pleopods and gills.

  3. Introduction to application of spot welding sealants in manufacture of white body%浅谈点焊密封胶在白车身制造中的应用

    王晓宁

    2013-01-01

    The spot welding sealant should be applied within the welding joints to ensure the leak and air tightness of the finished automobiles and meet the increasingly stringent requirements of NVH and rust prevention.The classification,using process and performance of the spot welding sealants were introduced.The actual production experience showed that the selection of suitable spot welding sealants,curing process and sealant spreading has an important role to assure the quality and performance of the finished aotomobiles.%为保证整车的密封性,满足日益严格的NVH与防锈要求,需在焊缝间涂布点焊密封胶。介绍了点焊密封胶分类、使用工艺及性能。实际生产经验表明,选择合适的点焊密封胶、固化工艺及涂胶量,对保证整车品质和性能具有重要作用。

  4. 渗透树脂治疗固定矫治后前牙唇面白垩斑的临床观察%Clinical observation on the effect of infiltration resin for the labial white spot lesions of anterior teeth after orthodontic treatment

    杨红丽; 唐睿; 黄珊珊; 杨雁

    2016-01-01

    目的:观察渗透树脂治疗固定矫治后前牙唇面白垩斑的临床效果。方法:选取32例179颗患牙,参照ICDAS评估方法分为A、B、C组,观察治疗后即刻、1周及1年的治疗效果。结果:经ANOVA及Duncan多重比较,3组患牙疗效具有极显著差异(P<0.01),术前、术后1周及术后1年比较显示治疗效果不会随时间变化产生显著差异(P=0.91)。结论:渗透树脂治疗能改善患牙的美观,临床效果与患牙的病损程度有关,治疗1年后牙面颜色稳定。ICDAS方法是简单、有效、易掌握的评估方法。%Objective:To evaluate the effect of infiltration resin on the labial white spot lesions of anterior teeth after orthodontic treatment. Method:179 teeth with white spot lesions from 32 patient,and divided into A,B,C groups based on ICDAS criterion,and treated with infiltration resin. The teeth were examined immediate,one week and one year after the treatment. Result:There were significant difference among the three groups after infiltration resin treatment (ANOVA and Duncan test,P<0.01). The difference were insignificant before,one week or one year after the treatment(P=0.91). Conclu-sion:The infiltration resin treatment was effective for the white spot lesion at the early caries and could enhance the aesthetic appearance,which was stable after one-year recall. This investigation demonstrates ICDAS criterion as both a highly clinic-feasible and efficient method for caries grading.

  5. Meteorology of Jupiter's Equatorial Hot Spots and Plumes from Cassini

    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...

  6. Modulatory effect of CpG oligodeoxynucleotide on a DNA vaccine against nervous necrosis virus in orange-spotted grouper (Epinephelus coioides).

    Chen, Shiang-Peng; Peng, Ran-Hong; Chiou, Pinwen P

    2015-08-01

    We report the development of a DNA vaccine pcMGNNV2 against nervous necrosis virus (NNV), a leading cause of mass mortality in grouper larvae. In addition, the modulatory effect of CpG oligodeoxynucleotide (ODN), a Toll-like receptor 9 agonist, on the DNA vaccine was evaluated. The DNA vaccine alone elicited the production of NNV-specific antibodies, indicating that the vaccine was capable of triggering adaptive humoral response. Furthermore, significant induction of TLR9, Mx and IL-1β was observed in the spleen on day 7 post-vaccination, supporting that the vaccine could trigger TLR9 signaling. The incorporation of CpG ODN at high dose did not significantly affect the level of NNV-specific antibodies, but was able to moderately enhance the expression of Mx and IL-1β on day 7, indicating its ability in modulating innate response. After challenge with NNV, the vaccine alone enhanced the survival rate in infected larvae at both 1 and 2 weeks post-vaccination. The combination of CpG ODN further increased the survival rate at week 1 but not week 2. Interestingly, at week 2 the ODN appeared to induce a Th1-like response, as indicated by upregulation of T-bet (a Th1 marker) and downregulation of GATA-3 (a Th2 marker). Thus, the results suggest that the boosted Th1 response by CpG ODN does not augment the protection efficacy of pcMGNNV2 vaccine. To our best knowledge, this is the first report of a successful DNA vaccine against NNV in grouper.

  7. Spotted Seal Distribution Map

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

  8. Mononucleosis spot test

    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. Identification and Sequence Analysis of Tembusu Virus Isolated from White-feather Mule Ducks%白羽半番肉鸭坦布苏病毒的分离鉴定及结构蛋白基因分析

    傅光华; 陈红梅; 黄瑜; 万春和; 傅秋玲; 程龙飞; 施少华; 林芳; 林建生

    2012-01-01

    A viral isolate (named as ZZ40) was abtained from liver of white-feather mule ducks with irregular hemorrhagic hepatitis, and were identified as duck tembus virus (DTMUV) by RT-PCR. The virus was specifically neutralized by anti-DTMUV hyperimmune serum. Gene fregments were abtained by using three sets of primers aimed at viral structural genes, and shared no less than 99.1% nucleotide sequence identity with those of avian tembus viruses isolated recently, and no more than 88. 6% with that of Mosquito-borne tembus virus MM-1775 strain. Phylogenetic analysis showed that ZZ40 strain clustal into the same clade with those duck viruses isolated recently. These results indicated that the white-feather mule ducks in this case was infected with duck tembusu virus, which has no abviously variation in antigenicity compared with those of the early isolates of duck tembusu virus from egg-laying duck.%自肝脏表现不规则出血的白羽半番肉鸭脏器中分离获得1株病毒(命名为ZZ40株),经(RT-) PCR检测为鸭坦布苏病毒,可被鸭坦布苏病毒高免阳性血清特异性中和.应用鸭坦布苏病毒特异性引物从该株病毒基因组中成功获得结构蛋白基因片段,该片段与近年来我国分离的禽源坦布苏病毒株核酸序列同源性均在99.1%以上,而与蚊媒源坦布苏病毒MM-1775株同源性仅为88.6%;分子进化分析表明该株病毒与BYD-1等近年来的鸭坦布苏病毒分离株共处于一个进化分支.以上结果表明,我国白羽半番肉鸭存在坦布苏病毒感染,且该分离毒株的抗原性与我国早期分离于种(蛋)鸭的坦布苏病毒差异不明显.

  10. The role of shrimp miR-965 in virus infection.

    Shu, Le; Li, Changrun; Zhang, Xiaobo

    2016-07-01

    RNAi, mediated by microRNAs (miRNAs), has attracted increasing attention for its important role in cross-talk between host and virus. However, the role of host miRNA in the virus infection in vivo has not been intensively investigated. In this study, the effects of a shrimp miRNA (miR-965) on the white spot syndrome virus (WSSV) infection were characterized. The results indicated that the expression of miR-965 was significantly upregulated in shrimp in response to the WSSV challenge, suggesting its involvement in the virus infection. The miR-965 silencing led to significant increases of WSSV copies and virus-infected shrimp mortality, while the miR-965 overexpression resulted in the decreased WSSV copies and virus-infected shrimp mortality, indicating that miR-965 played a negative role in the WSSV infection. The further data revealed that miR-965 inhibited the virus infection by targeting the viral wsv240 gene, an important gene required for the WSSV infection in shrimp. The results demonstrated that miR-965 could promote the shrimp phagocytosis against virus infection by targeting the shrimp ATG5 (autophagy related 5) gene. Therefore, our findings presented novel evidence to better understand the anfractuous host-virus interactions in vivo.

  11. Efficacy of CMX001 as a Post Exposure Antiviral in New Zealand White Rabbits Infected with Rabbitpox Virus, a Model for Orthopoxvirus Infections of Humans

    Scott Foster

    2011-01-01

    Full Text Available CMX001, a lipophilic nucleotide analog formed by covalently linking 3-(hexdecyloxypropan-1-ol to cidofovir (CDV, is being developed as a treatment for smallpox. In the absence of human cases of smallpox, new treatments must be tested for efficacy in animal models. Previously, we demonstrated the efficacy of CMX001 in protecting New Zealand White rabbits from mortality following intradermal infection with rabbitpox virus as a model for smallpox, monkeypox and for treatment of adverse reactions to smallpox vaccination. Here we extend these studies by exploring different dosing regimens and performing randomized, blinded, placebo-controlled studies. In addition, because rabbitpox virus can be transmitted via naturally generated aerosols (animal to animal transmission, we report on studies to test the efficacy of CMX001 in protecting rabbits from lethal rabbitpox virus disease when infection occurs by animal to animal transmission. In all cases, CMX001 treatment was initiated at the onset of observable lesions in the ears to model the use of CMX001 as a treatment for symptomatic smallpox. The results demonstrate that CMX001 is an effective treatment for symptomatic rabbitpox virus infection. The rabbitpox model has key similarities to human smallpox including an incubation period, generalized systemic disease, the occurrence of lesions which may be used as a trigger for initiating therapy, and natural animal to animal spread, making it an appropriate model.

  12. Identification of Tomato spotted wilt virus and Screening for Resistant Sources in Tomato%番茄斑萎病毒TSWV的鉴定及抗病种质的筛选

    邱树亮; 王孝宣; 杜永臣; 高建昌; 国艳梅; 朱德蔚; 胡鸿; 李宝聚; 石延霞

    2012-01-01

    Fifteen tomato plants which seemed infected Tomato spotted wilt virus (TSWV) were selected randomly from the tomato cultivation greenhouse of Shunyi District, Beijing. Then the leaves and fruits were obtained from each plant and tested for the presence of TSWV using a DAS-ELISA kit. The results showed positive, indicating that 15 tomato materials had been infected with TSWV. A genetic analysis of 442 tomato materials with co-dominant SCAR markers developed from Sw-5, TSWV resistance gene originated from Solanum peruvianum, was issued and identified 24 resistant materials, which including 4 wild tomatoes, 1 hybrid, 16 breeding inbred lines and 3 S. pennellii introgression lines. Those 4 wild tomatoes are not suitable as breeding materials because of the poor agronomic characters, i.e. small fruits, low production, poor quality and bad flavor, while the other 20 tomato materials can be used for developing cultivars resistant to TSWV due to their better agronomic characters.%在北京顺义地区番茄栽培温室中随机抽取了15份疑似感染番茄斑萎病毒(Tswv)的植株叶片和果实,利用TSwv试剂盒进行DAS-ELISA检测,结果均为阳性,表明其感染了Tswv。利用来自秘鲁番茄的抗TSWV,基因Sw-5的共显性SCAR标记,对442份番茄材料进行分析,筛选出24份含有Sw-5的材料,其中4份为野生番茄,1份为杂交种,16份为高代育种自交系,3份为野生潘那利番茄渐渗系。4份野生番茄综合农艺性状较差,如果实小、产量低、品质和风味差等,不能直接应用于育种;其余20份含Sw-5的材料综合农艺性状较好,可作为番茄抗TSWV的育种材料。

  13. Prevalence of Three Shrimp Viruses in Zhejiang Province in 2008

    Xiao-wei Yu; Jian-ping Wang; Wei Zhang; Zheng-li Shi

    2011-01-01

    White spot syndrome virus(WSSV), Taura syndrome virus(TSV)and Infectious hypodermal and haematopoietic necrosis virus(IHHNV)are three shrimp viruses responsible for major pandemics affecting the shrimp farming industry. Shrimps samples were collected from 12 farms in Zhejiang province, China, in 2008 and analyzed by PCR to determine the prevalence of these viruses. From the 12 sampling locations, 8 farms were positive for WSSV, 8 for IHHNV and 6 for both WSSV and IHHNV. An average percentage of 57.4% of shrimp individuals were infected with WSSV, while 49.2% were infected with IHHNV. A high prevalence of co-infection with WSSV and IHHNV among samples was detected from the following samples: Bingjiang(93.3%), liuao(66.7%), Jianshan(46.7%)and Xianxiang(46.7%). No samples exhibited evidence of infection with TSV in collected samples. This study provides comprehensive information of the prevalence of three shrimp viruses in Zhejiang and may be helpful for disease prevention control in this region.

  14. Development of a Liquid Chip Technique to Simultaneously Detect Taura Syndrome Virus( TSV) and Yellow Head Disease Virus( YHDV)

    Yin; Weili; Zhang; Sihua; Yue; Zhiqin; Zheng; Xiaolong; Liu; Hong

    2014-01-01

    The aim is to develop a liquid chip technique to detect Taura syndrome virus( TSV) and yellow head disease virus( YHDV) on Penaeus orientalis simultaneously. The CP2 gene of TSV and N gene of YHDV in Gen Bank was analysed by using the software DNAStar 7. 0 to design the TSV-and YHDV-specific primers. The primers were labeled with biotin and subjected to amination modification. They were then coupled with fluorescence-coded microspheres and then used for hybridization with RT- PCR products of TSV and YHDV. The liquid chip detection technique for detection of TSV and YHDV was established by using BD FACSArray to detect fluorescence signal in the reaction system. This assay system had a high sensitivity to TSV and YHDV,with the detection of limit of 100 pg. Moreover,the assay was specific for the detection of TSV,YHDV and was not susceptible to cross with other viruses,including white spot syndrome virus( WSSV),spring viremia of carp virus( SVCV),infectious haematopoietic necrosis virus( IHNV). In conclusion,the liquid chip assay technique established in this study is highly sensitive and specific to TSV and YHDV detection. Moreover,it provides a novel,convenient and rapid approach for the detection of TSV and YHDV.

  15. 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

    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

  16. SpotADAPT

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

    2015-01-01

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

  17. Arc spot grouping: An entanglement of arc spot cells

    Kajita, Shin, E-mail: kajita.shin@nagoya-u.jp [EcoTopia Science Institute, Nagoya University, Nagoya 464-8603 (Japan); Hwangbo, Dogyun; Ohno, Noriyasu [Graduate School of Engineering, Nagoya University, Nagoya 464-8603 (Japan); Tsventoukh, Mikhail M. [Lebedev Physical Institute, Russian Academy of Sciences, Moscow 119991 (Russian Federation); Barengolts, Sergey A. [Prokhorov General Physics Institute, Russian Academy of Sciences, Moscow 119991 (Russian Federation)

    2014-12-21

    In recent experiments, clear transitions in velocity and trail width of an arc spot initiated on nanostructured tungsten were observed on the boundary of the thick and thin nanostructured layer regions. The velocity of arc spot was significantly decreased on the thick nanostructured region. It was suggested that the grouping decreased the velocity of arc spot. In this study, we try to explain the phenomena using a simple random walk model that has properties of directionality and self-avoidance. And grouping feature was added by installing an attractive force between spot cells with dealing with multi-spots. It was revealed that an entanglement of arc spot cells decreased the spot velocity, and spot cells tend to stamp at the same location many times.

  18. SURVEY OF VIRAL DISEASES OF PACIFIC WHITE SHRIMP, Litopenaeus vannamei IN INDONESIA

    Taukhid Taukhid

    2008-06-01

    Full Text Available Penaeid shrimp culture is a major contributor to foreign exchange earning in Indonesia. It has significant impact on economic development of fisheries sector, and leads to be one of prime mover to improve social prosperity. However, shrimp industry particularly black tiger shrimp (Penaeus monodon has been facing unpredictable situation due to disease problem. The main constrain in correlation to the development of shrimp industry is disease outbreak, especially caused by viral agents. White Spot Syndrome Virus (WSSV occurred in 1994, causing mass mortality of black tiger shrimp almost in all of the middle and western part of Indonesia. Due to the disease problem, it is estimated that in year 2000, more than 50% of shrimp pond were idle. Pacific white shrimp (Litopenaeus vannamei or “udang vanamei” was introduced to Indonesia at the end of 1999, and released officially in July, 2001. Response of shrimp farmers to the shrimp rapidly accepted and distributed to many provinces in the country. At the end of 2006, distribution of white shrimp culture was encountered in more than 15 provinces. The seeds are mainly produced from hatcheries located in East Java and Lampung. The information of TSV in Indonesia was reported firstly from East Java at the end of 2002, without a clear history. Since then, survey of TSV distribution was conducted intensively in white shrimp production areas. Beside TSV, population of white shrimp coming to Indonesia also susceptible to White Spot Syndrome Virus (WSSV and Infectious Hypodermal and Hematopoietic Necrosis Virus (IHHNV infection. A survey with the aim to know significant viral diseases of white shrimp is needed to set up an alternative strategy to control them. The survey was conducted, firstly in the main production centers of white shrimp; and planned to be continued throughout the country. Samples collection, diagnostic method and data compiled in this study were collected from both

  19. Meteorology of Jupiter's Equatorial Hot Spots and Plumes from Cassini

    Choi, David Sanghun; Showman, Adam P.; 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 relatively cloud-free regions that emit strongly at 5 lm; improved knowledge of these features is crucial for fully understanding Galileo probe measurements taken during its descent through one. Hot spots are quasistable, 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 cirrus-like 'scooter' clouds flow rapidly through the plumes before disappearing within the dark areas. These clouds travel at 150-200 m/s, much faster than the 100 m/s hot spot and plume drift speed. This raises the possibility that the scooter clouds may be more illustrative of the actual jet stream speed at these latitudes. Most previously published zonal wind profiles represent the drift speed of the hot spots at their latitude from pattern matching of the entire longitudinal image strip. If a downward branch of an equatorially-trapped Rossby wave controls the overall appearance of hot spots, however, the westward phase velocity of the wave leads to underestimates of the true jet stream speed.

  20. A New Definition of Virus

    HE Hongjun; CAO Sihua; LUO Li; FENG Tao; PAN Li; ZOU Zhiji

    2006-01-01

    Security experts have not formally defined the distinction between viruses and normal programs. The paper takes user's intension as the criteria for malice, gives a formal definition of viruses that aim at stealing or destroying files, and proposes an algorithm to detect virus correctly. Compared with traditional definitions, this new definition is easy to understand, covers more malwares, adapts development of virus technology, and defines virus on the spot. The paper has also analyzed more than 250 real viruses and finds that they are all in the domain of the new definition, this implies that the new definition has great practical significance.

  1. Spot- Zombie Filtering System

    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.

  2. El spot electoral negativo

    Palma Peña-Jiménez

    2011-01-01

    Full Text Available l spot político tiene durante la campaña un objetivo final inequívoco: la consecución del voto favorable. Se dirige al cuerpo electoral a través de la televisión y de Internet, y presenta, en muchos casos, un planteamiento negativo, albergando mensajes destinados a la crítica frontal contra el adversario, más que a la exposición de propuestas propias. Este artículo se centra en el análisis del spot electoral negativo, en aquellas producciones audiovisuales construidas sin más causa que la reprobación del contrincante. Se trata de vídeos que, lejos de emplearse en difundir las potencialidades de la organización y las virtudes de su candidato –además de su programa electoral–, consumen su tiempo en descalificar al oponente mediante la transmisión de mensajes, muchas veces, ad hominem. Repasamos el planteamiento negativo del spot electoral desde su primera manifestación, que en España data de 1996, año de emisión del conocido como vídeo del dóberman, sin olvidar otros ejemplos que completan el objeto de estudio.

  3. Spot- Zombie Filtering System

    Arathy Rajagopal

    2015-10-01

    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.

  4. [Molecular-genetic characterization of the Okhotskiy virus (OKHV) and Aniva virus (ANIV) (Orbivirus, Reoviridae) isolated from the ticks Ixodes (Ceratixodes) uriae White, 1852 in high latitudes of the Northern Eurasia].

    L'vov, D K; Al'khovskiĭ, S V; Shchelkanov, M Iu; Shchetinin, A M; Deriabin, P G; Gitel'man, A K; Samokhvalov, E I; Botikov, A G

    2014-01-01

    Molecular-genetic characteristics of the Okhotskiy virus (OKHV) and Aniva virus (ANIV) were studied (ID GenBank KF981623-32). These viruses are distributed over the shelf and Island areas in the high latitudes in the Okhotsk, Bering, and Barents seas and linked with nesting colonies of Alcidae seabirds and their obligatory parasites, the Ixodes uriae (Ixodidae) ticks. OKHV and ANIV are observed to be independent species within the limits of the Great Island virus (GIV) group of the Orbivirus genus of the Reoviridae family. The majority of the genes of OKHV and ANIV have high homology (VP1 - 96%, T2 - 99%, VP7 (T13) - 98%, NS1 - 94%, NS2 - 98%, NS3 - 72%, VP6 - 93%). Nevertheless, the envelope proteins containing the main specific antigenic determinants (VP2 and VP5) of OKHV and ANIV are sufficiently different (62% and 68% homology for amino acid sequences, respectively).

  5. Ceres' Yellow Spots - Observations with Dawn Framing Camera

    Schäfer, Michael; Schäfer, Tanja; Cloutis, Edward A.; Izawa, Matthew R. M.; Platz, Thomas; Castillo-Rogez, Julie C.; Hoffmann, Martin; Thangjam, Guneshwar S.; Kneissl, Thomas; Nathues, Andreas; Mengel, Kurt; Williams, David A.; Kallisch, Jan; Ripken, Joachim; Russell, Christopher T.

    2016-04-01

    The Framing Camera (FC) onboard the Dawn spacecraft acquired several spectral data sets of (1) Ceres with increasing spatial resolution (up to 135 m/pixel with nearly global coverage). The FC is equipped with seven color filters (0.4-1.0 μm) plus one panchromatic ('clear') filter [1]. We produced spectral mosaics using photometrically corrected FC color filter images as described in [2]. Even early FC color mosaics obtained during Dawn's approach unexpectedly exhibited quite a diversity of surface materials on Ceres. Besides the ordinary cerean surface material, potentially composed of ammoniated phyllosilicates [3] or some other alteration product of carbonaceous chondrites [4], a large number of bright spots were found on Ceres [5]. These spots are substantially brighter than the average surface (exceeding its triple standard deviation), with the spots within Occator crater being the brightest and most prominent examples (reflectance more than 10 times the average of Ceres). We observed bright spots which are different by their obvious yellow color. This yellow color appears both in a 'true color' RGB display (R=0.65, G=0.55, B=0.44 μm) as well as in a false color display (R=0.97, G=0.75, B=0.44 μm) using a linear 2% stretch. Their spectra show a steep red slope between 0.44 and 0.55 μm (UV drop-off). On the contrary to these yellow spots, the vast majority of bright spots appears white in the aforementioned color displays and exhibit blue sloped spectra, except for a shallow UV drop-off. Thus, yellow spots are easily distinguishable from white spots and the remaining cerean surface by their high values in the ratio 0.55/0.44 μm. We found 8 occurrences of yellow spots on Ceres. Most of them (>70 individual spots) occur both inside and outside crater Dantu, where white spots are also found in the immediate vicinity. Besides Dantu, further occurrences with only a few yellow spots were found at craters Ikapati and Gaue. Less definite occurrences are found at 97

  6. 青海湖裸鲤体表白点病病原菌的分离与药敏试验%Identification and Drug Sensitivity Test of Pathogenic Bacteria of White-spot Disease From Skin Surface of Gymnocypris przewaiskii

    李增魁; 李长忠; 康明; 圈华; 张勤文; 刘道鑫; 马东红; 夏若措毛; 李英

    2015-01-01

    To assure the pathogenic bacteria of white-spot disease from skin surface in Gymnocypris prze-waiskii and treat the disease,three bacterial strains (Citrobacter freundii ,Aeromonas veronii ,Morganella fulton)were isolated from naturally infected Gymnocypris przewaiskii and identified by morphology,cul-tural characteristics,physical and chemical characters,and 16 S rRNA sequence analysis.The three strains were Gram negative.Colony morphologies were consistent.Drug sensitivity test revealed that the three strains were highly sensitive to cefepime,amikacin and gentamicin,which will benefit the treatment of dis-ease in Gymnocypris przewaiskii .%为确定养殖青海湖裸鲤(Gymnocypris przewaiskii )体表白点病的病原,分别对患病鱼白色点状囊泡样物进行了细菌分离、生理生化特性鉴定、16 S rRNA 基因分析和药敏感试验,分离到了以弗氏柠檬酸杆菌(Citrobacter freundii )、维氏气单胞菌(Aeromonas veronii ,AV)和摩氏摩根菌(Morganella fulton )为代表的菌落形态一致、革兰阴性细菌;且分离到的3种菌都对头孢吡肟、丁胺卡那霉素、庆大霉素等药物敏感。

  7. Spots and White Light Flares in an L Dwarf

    2013-01-01

    Program GN-2012A-Q-37) GMOS spectrograph (Hook et al. 2004) when a series of flares occurred. A spectrum of the most powerful flare in its impulsive...10:14 Hα HeI HeI HeI OI Fig. 4. Gemini-North GMOS spectra of W1906+40 in quiescence (below) and in flare. Note the broad Hα, atomic emission lines

  8. 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

    A. S. Costa

    1957-01-01

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

  9. Turbulent spots in hypervelocity flow

    Jewell, Joseph S.; Leyva, Ivett A.; Shepherd, Joseph E.

    2017-04-01

    The turbulent spot propagation process in boundary layer flows of air, nitrogen, carbon dioxide, and air/carbon dioxide mixtures in thermochemical nonequilibrium at high enthalpy is investigated. Experiments are performed in a hypervelocity reflected shock tunnel with a 5-degree half-angle axisymmetric cone instrumented with flush-mounted fast-response coaxial thermocouples. Time-resolved and spatially demarcated heat transfer traces are used to track the propagation of turbulent bursts within the mean flow, and convection rates at approximately 91, 74, and 63% of the boundary layer edge velocity, respectively, are observed for the leading edge, peak, and trailing edge of the spots. A simple model constructed with these spot propagation parameters is used to infer spot generation rates from observed transition onset to completion distance. Spot generation rates in air and nitrogen are estimated to be approximately twice the spot generation rates in air/carbon dioxide mixtures.

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

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

    1994-01-01

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

  11. Deep Sequencing Analysis of Apple Infecting Viruses in Korea

    In-Sook Cho; Davaajargal Igori; Seungmo Lim; Gug-Seoun Choi; John Hammond; Hyoun-Sub Lim; Jae Sun Moon

    2016-01-01

    Deep sequencing has generated 52 contigs derived from five viruses; Apple chlorotic leaf spot virus (ACLSV), Apple stem grooving virus (ASGV), Apple stem pitting virus (ASPV), Apple green crinkle associated virus (AGCaV), and Apricot latent virus (ApLV) were identified from eight apple samples showing small leaves and/or growth retardation. Nucleotide (nt) sequence identity of the assembled contigs was from 68% to 99% compared to the reference sequences of the five respective viral genomes. S...

  12. The SPOT satellite

    Fouquet, J.-P.

    1981-03-01

    The background, objectives and data products of the French SPOT remote sensing satellite system are presented. The system, which was developed starting in 1978 with the subsequent participation of Sweden and Belgium, is based on a standard multimission platform with associated ground control station and a mission-specific payload, which includes two High-Resolution Visible range instruments allowing the acquisition of stereoscopic views from different orbits. Mission objectives include the definition of future remote sensing systems, the compilation of a cartographic and resources data base, the study of species discrimination and production forecasting based on frequent access and off-nadir viewing, the compilation of a stereoscopic data base, and platform and instrument qualification, for possible applications in cartography, geology and agriculture. Standard data products will be available at three levels of preprocessing: radiometric correction only, precision processing for vertical viewing, and cartographic quality processing.

  13. Divide and conquer spot noise

    Leeuw, W.C. de; Liere, R. van

    1997-01-01

    The design and implementation of an interactive spot noise algorithm is presented. Spot noise is a technique which utilizes texture for the visualization of flow fields. Various design tradeoffs are discussed that allow an optimal implementation on a range of high end graphical workstations. Two app

  14. Black-spot poison ivy.

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

    2008-01-01

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

  15. European Whiteness?

    Blaagaard, Bolette

    2008-01-01

    Born out of the United States’ (U.S.) history of slavery and segregation and intertwined with gender studies and feminism, the field of critical whiteness studies does not fit easily into a European setting and the particular historical context that entails. In order for a field of European...

  16. 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

    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

  17. Development of a novel baculovirus titration method using the Enzyme-linked immunosorbent spot (ELISPOT) assay.

    Wang, Wei; Cheng, Tong; Ma, Ke; Xia, Dezhen; Wang, Yongmei; Liu, Jian; Du, Hailian; Shih, James Wai Kuo; Zhang, Jun; Zhao, Qinjian; Xia, Ningshao

    2013-03-01

    The baculovirus expression vector system (BEVS) is one of the most powerful methods for production of recombinant proteins for research or commercial purposes. Titration of viable virus in insect cell culture is often required when BEVS is used for basic research or bioprocessing. An enzyme-linked immunosorbent spot (ELISPOT) assay using monoclonal antibodies against the major capsid protein VP39 of both Autographa californica nuclear polyhedrosis virus (AcMNPV) and Bombyx mori nuclear polyhedrosis virus (BmNPV) was developed for baculovirus quantitation at 48h post-infection. The titer was determined by visualizing infected insect cells as blue spots and automated spot counting was achieved with ELISPOT hardware and software. Log-scale comparison of the results between the ELISPOT assay and a conventional end point dilution assay using a fluorescent marker showed a good correlation for both AcMNPV (R(2)=0.9980, pspot counting.

  18. Coulomb explosion of "hot spot"

    Oreshkin, V I; Chaikovsky, S A; Artyomov, A P

    2016-01-01

    The study presented in this paper has shown that the generation of hard x rays and high-energy ions, which are detected in pinch implosion experiments, may be associated with the Coulomb explosion of the hot spot that is formed due to the outflow of the material from the pinch cross point. During the process of material outflow, the temperature of the hot spot plasma increases, and conditions arise for the plasma electrons to become continuously accelerated. The runaway of electrons from the hot spot region results in the buildup of positive space charge in this region followed by a Coulomb explosion. The conditions for the hot spot plasma electrons to become continuously accelerated have been revealed and estimates have been obtained for the kinetic energy of the ions generated by the Coulomb explosion.

  19. Development and evaluation of a loop-mediated isothermal amplification assay for rapid detection of lymphocystis disease virus.

    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.

  20. White Paranoia

    Jørholt, Eva

    2017-01-01

    Inspired by Alain Robbe-Grillet’s novel La Jalousie (1957), the essay contends that Michael Haneke’s Caché (2005) takes its viewers inside a postcolonial white paranoia which is, arguably, the root cause of the exclusion, segregation and racist discrimination that many immigrants from the former....... The construction of the film, however, also allows Haneke to critically comment upon his protagonist’s paranoia and demonstrate its ill-foundedness, for instance by pointing to a possible ‘banal conviviality’ (Paul Gilroy) between people of different ethnicities, cultures and religions....