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1
Franks, David. "Occurrence and Evaluation of White Spot Lesions in Orthodontic Patients: A Pilot Study." Master's thesis, Temple University Libraries, 2014. http://cdm16002.contentdm.oclc.org/cdm/ref/collection/p245801coll10/id/286676.
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Oral BiologyM.S.
Orthodontic treatment may cause an increase in the rate of enamel decalcification on tooth surfaces, producing White Spot Lesions (WSL). Orthodontic patients are at a higher risk for decalcification because orthodontic appliances retain food debris which leads to increased plaque formation. Dental plaque, an oral biofilm formed by factors including genetics, diet, hygiene, and environment, contains acid producing bacterial strains with a predominance of Mutans Streptococcus (MS). MS and others metabolize oral carbohydrates during ingestion, the byproducts of which acidify the biofilm to begin a process of enamel decalcification and formation of WSL. This study tests if patients in orthodontic treatment at Temple University can be used as subjects for further longitudinal study of WSL risk factors. Twenty patients between the ages of ten to eighteen after three months or greater of treatment were enrolled to determine if duration of treatment, hygiene, sense of coherence, obesity, diet frequencies, age and gender correlated with development of WSL. Of these, age is positively correlated with the number of untreated decayed surfaces. WSL and plaque levels may negatively correlate with increased brushing frequency and duration, while flossing frequency demonstrated a statistically significant negative correlation. This population may be suitable for further study because of its high incidence of WSL (75%), however difficulty in enrollment and patient attrition necessitates that future studies be modified.
Temple University--Theses
2
Dixon, Julian. "Prevalence of White Spot Lesions during Orthodontic Treatment." VCU Scholars Compass, 2009. http://scholarscompass.vcu.edu/etd/1843.
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The reported prevalence of decalcification in orthodontic patients varies from 2 to 96% mainly due to the lack of a standard examination technique. The aims of this study were: 1) to determine the prevalence of white spot lesions around brackets using visual examination and the DIAGNOdent; 2) to determine which teeth were the most susceptible to decalcification; and 3) to test the accuracy of the DIAGNOdent by comparing to the visual examination. The presence of white spot lesions was determined in two groups of patients who were 6 and 12 months into orthodontic treatment, respectively. The control group consisted of patients who were examined for white spot lesions immediately after having their braces placed on their teeth. The prevalence of white spot lesions was 38%, 46%, and 11% for the 6-month, 12-month, and control groups, respectively. There was a statistically significant correlation (r = 0.71) between the DIAGNOdent measurements and the visual examination.
3
Wulc, Daniel. "Treatment of Orthodontic White Spots: Etiology of Orthodontic White Spot Lesions and Interventional Fluoride Varnish Treatment: A Randomized Control Trial." Master's thesis, Temple University Libraries, 2015. http://cdm16002.contentdm.oclc.org/cdm/ref/collection/p245801coll10/id/328850.
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Oral BiologyM.S.
Fixed orthodontic appliances harbor plaque and perpetuate the formation of early non-cavitated carious lesions. However, not all patients with poor hygiene develop them. It has been established that fluoride varnish can be used to promote enamel remineralization. The study aimed to assess the efficacy of fluoride varnish in remineralizing early non-cavitated lesions among orthodontic patients. A second goal of this study was to elucidate if BMI and obesity increased susceptibility to development of white spot lesions. A randomized control trial was conducted among 25 patients attending the Orthodontic clinic at Temple University. Patients were ages 11-18 and had fixed orthodontic appliances for a minimum of three months. Eleven were randomly assigned to a test group (Enamel Pro® Varnish fluoride varnish application to white spot lesions every two months) and 14 randomly assigned to a control group (reinforcement of oral hygiene instructions). Data collection was completed every two months over a six-month time period. White spot lesion size was measured using the International Caries Detection and Assessment System (ICDAS). Oral hygiene was assessed using Plaque Index (PI) and S. mutans levels were measured using Stripmutans plaque/salivary tests (Dentocult®). Both the control and experimental group had non-significant decreases in non-cavitated carious lesion count. The control group displayed significant increases in Stripmutans salivary scores (p0.05). PI scores decreased in the control group and increased in the experimental group (p>0.05). There was no correlation between BMI and lesion count in the control or experimental group (p>0.05). A 5% sodium fluoride varnish containing Amorphous Calcium Phosphate (Enamel Pro® Varnish) fluoride varnish application was not efficacious in reducing early non-cavitated carious lesions when compared to reinforcing oral hygiene. There is no correlation with BMI and white spot susceptibility.
Temple University--Theses
4
Jiravanichpaisal, Pikul. "White Spot Syndrome Virus Interaction with a Freshwater Crayfish." Doctoral thesis, Uppsala : Acta Universitatis Upsaliensis : Univ.-bibl. [distributör], 2005. http://urn.kb.se/resolve?urn=urn:nbn:se:uu:diva-5776.
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You, Zerong. "A multifaceted study of White Spot Syndrome Virus (WSSV) a shrimp pathogen." Thesis, University of Hawaii at Manoa, 2003. http://proquest.umi.com/pqdweb?index=0&did=765959821&SrchMode=1&sid=1&Fmt=2&VInst=PROD&VType=PQD&RQT=309&VName=PQD&TS=1208557872&clientId=23440.
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Maxfield, Blake. "Perceived Responsibility for the Development of White Spot Lesions during Orthodontic Treatment." VCU Scholars Compass, 2009. http://scholarscompass.vcu.edu/etd/1842.
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White spot lesions (WSLs) or decalcifications remain a common complication in orthodontic patients with poor oral hygiene. The purpose of this study was to compare attitudes regarding the development of WSLs among patients, parents, orthodontists and general dentists and improve prevention and treatment protocols through better communication. A survey was developed to evaluate and compare the current opinions of orthodontic patients (n=315), parents (n=279), orthodontists (n=305) and general dentists (n=191) regarding the significance, prevention and treatment of WSLs. All four groups indicated that WSLs did detract from the overall appearance of straight teeth. All four groups indicated that patients were the most responsible for the prevention of WSLs. All four groups indicated that the general dentist should be more responsible for the treatment of WSLs than the orthodontist. General dentists were significantly more likely to indicate that the orthodontist was most responsible for the prevention of WSLs (P <0.005).
7
Inman, Alan John. "The biology and epidemiology of white leaf spot (Pseudocercosporella capsellae) on oilseed rape." Thesis, Queen Mary, University of London, 1992. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.411685.
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8
Enaia, Mahmoud [Verfasser]. "White spot lesions during multibracket appliance treatment : a challenge for clinical excellence / Mahmoud Enaia." Gießen : Universitätsbibliothek, 2011. http://d-nb.info/1063110416/34.
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9
Podray, Susan. "Current Technology and Techniques in Re-mineralization of White Spot Lesions: A Systematic Review." Master's thesis, Temple University Libraries, 2012. http://cdm16002.contentdm.oclc.org/cdm/ref/collection/p245801coll10/id/170366.
Full textAbstract:
Oral BiologyM.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 remineralization. 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 remineralization. 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.
Temple University--Theses
10
Bergdoll, Allison S. "Icon caries infiltrant resin and MI Paste Plus for the treatment of white spot lesions." Thesis, Birmingham, Ala. : University of Alabama at Birmingham, 2010. https://www.mhsl.uab.edu/dt/2010m/bergdoll.pdf.
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Al-Khateeb, Susan. "Studies on the remineralization of white spot lesions : longitudinal assessment with quantitative light-induced fluorescence /." Stockholm, 1998. http://diss.kib.ki.se/1998/19980331alkh.
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12
Corsin, Flavio. "An epidemiological study of white spot disease in the rice-shrimp farming system of Vietnam." Thesis, University of Liverpool, 2001. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.390630.
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13
Clark, Kristin Dumboski. "The efficacy of 37% phosphoric acid + Mi Paste Plus on remineralization of enamel white spot lesions." Thesis, University of Iowa, 2011. https://ir.uiowa.edu/etd/938.
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Purpose: This in vitro study evaluated the effectiveness of using a 37% phosphoric acid liquid etchant along with MI Paste Plus™ powered technology compared to using MI Paste Plus™ alone or to an artificially created saliva solution in decreasing the demineralization and enhancing the remineralization of artificial carious lesions created on extracted human teeth. The teeth were analyzed and compared using polarized light microscopy, quantitative light-induced fluorescence, and digital photography.
Materials and Methods: One hundred three recently extracted non-carious human third molar teeth without observable white-spot lesions, decalcification, or dental fluorosis were selected for this twelve day study and randomly divided into four treatment groups as follows:
Group 1 (Control) - Artificial saliva solution (27 teeth)
Group 2 (MIP) - MI Paste Plus™ application for 30 minutes daily for 12 days (26 teeth)
Group 3 (15MIP) - 15 second etch every third day and MI Paste Plus™ application for 30 minutes daily for 12 days (25 teeth)
Group 4 (1MIP) - 1 minute etch on day one ONLY and application of MI Paste Plus™ for 30 minutes daily for 12 days (25 teeth).
Results: Results of one<–>way ANOVA revealed there was a significant effect for the type of treatment on the lesion depth (p = 0.0027). The post-hoc Tukey-Kramer's test indicated there was a statistically significant difference between the two groups (15MIP and 1MIP) that incorporated an acid etch in combination with MI Paste Plus™ and the group with exposure to MI Paste Plus™ alone (MIP). In addition, results of one<–>way ANOVA showed that there was no statistically significant effect for type of treatment on the change in fluorescence (p = 0.1417) or the change in density (p = 0.1934).
Conclusions: The results of the present study revealed there was a significant effect for the type of treatment on the lesion depth (p = 0.0027). However, the only significant difference found was between the two groups (15MIP and 1MIP) that incorporated an acid etch in combination with MI Paste Plus™ and the group with exposure to MI Paste Plus™ alone (MIP). Thus, daily applications of MI Paste Plus™, with or without an acid etch, did not produce a statistically significant difference in mean lesion depth when compared to the control (artificial saliva group). In addition, the results of the present study showed that there was no statistically significant effect for type of treatment on the change in fluorescence (p = 0.1417) or the change in density (p = 0.1934). Further research is needed to evaluate MI Paste Plus™ capability in prevention of demineralization and/or enhancement of remineralization by conducting randomized clinical trials.
14
Sarkhouh, Shaima Mansour. "Investigating the ultrastructure of enamel white spot lesions (WSL) using Optical Coherence Tomography at different length scales." Thesis, University College London (University of London), 2017. http://discovery.ucl.ac.uk/10040047/.
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White spot lesion (WSL) is the clinical presentation of early caries, which is a demineralisation that occurs at subsurface level, with a well-mineralised surface layer enclosing the lesion. Early diagnosis and treatment of WSL is crucial to prevent further destruction of tooth structure. The aim of this research is to investigate the potential of optical coherence tomography (OCT) to be used as an adjunct diagnostic clinical tool to evaluate the severity of such lesions. This research also compared the OCT outputs with traditional histology, X-ray Microtomography (XMT), Synchrotron X-ray Diffraction (SXRD) and Scanning Electron microscope (SEM). All specimens were collected from patients undergoing dental treatment at Eastman Dental Hospital with informed consent following ethical approvall. Initially, Artificial WSLs were induced on sound enamel surfaces using a buffered methylcellulose gel system at pH 4.6 for 7 and 14 days. Type-matched native WSL and healthy control teeth were selected based on ICDAS for comparison. Imaging of samples was obtained using OCT of whole teeth and by polarised microscopy, SXRD, XMT and SEM of polished 250 μm thick sections. Polarised microscope, XMT and SEM confirmed the findings of the OCT results. Images showed that the more back scattered signals recorded, the deeper the destruction throughout enamel thickness. SXRD results showed changes in enamel texture, which was interpreted from measuring crystallite orientations and lattice parameter. SXRD result showed some correlation with OCT images, however more investigation is required to confirm the findings. In conclusion, the variations observed in the back-scattered light in OCT experiment were because of mineral density variation within enamel structure, as well as the changes in prismatic structure and may be related to crystallite texture and orientation. OCT has shown to be a reliable non-destructive technique, that can investigate the internal structure, by measuring the back-scattered light from materials such as enamel and dentine. In healthy samples, OCT B-scans showed a homogenous pattern of scattering intensity throughout enamel structure, indicating healthy structure, while in both natural and induced white spot lesions, a non homogenous scattering intensity was observed, indicating changes in enamel structure.
15
Verbruggen, Bas. "Generating genomic resources for two crustacean species and their application to the study of White Spot Disease." Thesis, University of Exeter, 2016. http://hdl.handle.net/10871/25535.
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Over the last decades the crustacean aquaculture sector has been steadily growing, in order to meet global demands for its products. A major hurdle for further growth of the industry is the prevalence of viral disease epidemics that are facilitated by the intense culture conditions. A devastating virus impacting on the sector is the White Spot Syndrome Virus (WSSV), responsible for over US $10 billion in losses in shrimp production and trade. The Pathogenicity of WSSV is high, reaching 100 % mortality within 3-10 days in penaeid shrimps. In contrast, the European shore crab Carcinus maenas has been shown to be relatively resistant to WSSV. Uncovering the basis of this resistance could help inform on the development of strategies to mitigate the WSSV threat. C. maenas has been used widely in studies on ecotoxicology and host-pathogen interactions. However, like most aquatic crustaceans, the genomic resources available for this species are limited, impairing experimentation. Therefore, to facilitate interpretations of the exposure studies, we first produced a C. maenas transcriptome and genome scaffold assembly. We also produced a transcriptome for the European lobster (Homarus gammarus), an ecologically and commercially important crustacean species in United Kingdom waters, for use in comparing WSSV responses in this, a susceptible species, and C. maenas. For the C. maenas transcriptome assembly we isolated and pooled RNA from twelve different tissues and sequenced RNA on an Illumina HiSeq 2500 platform. After de novo assembly a transcriptome encompassing 212,427 transcripts was produced. Similar, the H. gammarus transcriptome was based on RNA from nine tissues and contained 106,498 transcripts. The transcripts were filtered and annotated using a variety of tools (including BLAST, MEGAN and RSEM) and databases (including GenBank, Gene Ontology and KEGG). The annotation rate for transcripts in both transcriptomes was around 20-25 % which appears to be common for aquatic crustacean species, as a result of the lack of well annotated gene sequences for this clade. Since it is likely that the host immune system would play an important role in WSSV infection we characterized the IMD, JAK/STAT, Toll-like receptor and other innate immune system pathways. We found a strong overlap between the immune system pathways in C. maenas and H. gammarus. In addition we investigated the sequence diversity of known WSSV interacting proteins amongst susceptible penaeid shrimp/lobster and the more resistant C. maenas. There were differences in viral receptor sequences, like Rab7, that correlate with a less efficient infection by WSSV. To produce the genome scaffold assembly for C. maenas we isolated DNA from muscle tissue and produced both paired-end and mate pair libraries for processing on the Illumina HiSeq 2500 platform. A de novo draft genome assembly consisting of 338,980 scaffolds and covering 362 Mb (36 % of estimated genome size) was produced, using SOAP-denovo2 coupled with the BESST scaffolding system. The generated assembly was highly fragmented due to the presence of repetitive areas in the C. maenas genome. Using a combination of ab initio predictors, RNA-sequencing data from the transcriptome datasets and curated C. maenas sequences we produced a model encompassing 10,355 genes. The gene model for C. maenas Dscam, a gene potentially involved in (pan)crustacean immune memory, was investigated in greater detail as manual curation can improve on the results of ab initio predictors. The scaffold containing C. maenas Dscam was fragmented, thus only contained the latter exons of the gene. The assembled draft genome and transcriptomes for C. maenas and H. gammarus are valuable molecular resources for studies involving these and other aquatic crustacean species. To uncover the basis of their resistance to WSSV, we infected C. maenas with WSSV and measured mRNA and miRNA expression for 7 time points spread over a period of 28 days, using RNA-Seq and miRNA-Seq. The resistance of C. maenas to WSSV infection was confirmed by the fact that no mortalities occurred. In these animals replicating WSSV was latent and detected only after 7 days, and this occurred in five of out 28 infected crabs only. Differential expression of transcripts and miRNAs were identified for each time point. In the first 12 hours post exposure we observed decreased expression of important regulators in endocytosis. Since it is established that WSSV enters the host cells through endocytosis and that interactions between the viral protein VP28 and Rab7 are important in successful infection, it is likely that changes in this process could impact WSSV infection success. Additionally we observed an increased expression of transcripts involved in RNA interference pathways across many time points, indicating a longer term response to initial viral exposure. miRNA sequencing showed several miRNAs that were differentially expressed. The most striking finding was a novel C. maenas miRNA that we found to be significantly downregulated in every WSSV infected individual, suggesting that it may play an important role in mediating the response of the host to the virus. In silico target prediction pointed to the involvement of this miRNA in endocytosis regulation. Taken together we hypothesize that C. maenas resistance to WSSV involves obstruction of viral entry by endocytosis, a process probably regulated through miRNAs, resulting in inefficient uptake of virions.
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Bateman, Kelly Simone. "Susceptibility of European crustaceans to White Spot Syndrome Virus (WSSV), a non-exotic, EC Directive-listed pathogen." Thesis, University of Southampton, 2014. https://eprints.soton.ac.uk/378999/.
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This project provides a definitive statement on the susceptibility of ecologically and economically important European crustacean species to White Spot Syndrome Virus (WSSV). Exposure trials revealed universal susceptibility to WSSV infection in seven hosts, and that relative susceptibility varies significantly between species. To determine whether WSSV infection would remain in the tissues of crabs as persistent infections (or whether these crabs would clear the viral infection from their systems over time) crabs were fed with WSSV-infected tissues and then observed in tanks for 3 months. Results suggested that the carcasses of infected (but not diseased) shore and edible crab crabs appear to pose a limited risk of transmission to susceptible hosts within the 3 month timeframe of the study. The European shore crab (Carcinus maenas) was shown to display a lower suscept ibility to White Spot Syndrome Virus (WSSV) when compared to other European decapod species. Despite showing signs of infection with WSSV, the shore crab appeared resistant to the development of disease and was highlighted as a possible asymptomatic carrier of the virus. Carcinus maenas individuals which had been injected with WSSV and then exposed to varying temperature stress conditions were compared. This suggested that crabs could be divided into two groups (high and low responders) according to differences in pathogenesis and viral replication. The response was not dependent on the presence or absence of an external stressor but was more likely an inherent capacity within individual crabs. A small scale survey of supermarket commodity shrimp confirmed the presence of WSSV-contaminated products within shrimp imported for human consumpt ion, demonstrating that frozen commodity shrimp is a route of entry for WSSV into Europe. Experimental trials showed that this virus is viable and that the commodity products were infective and could passage the infection to naive crustaceans that are known to be susceptible to the virus, highlighting this practice needs to be considered and controlled in risk assessments . Analysis of WSSV Variable Nucleotide Tandem Repeats (VNTR) within ORF 94 following passage through different hosts revealed subtle differences in WSSV VNTR types present in crab and crayfish tissues suggesting that the host may influence which viral type propagates during infections. These studies also identified and described two novel virus infections. The viral infections mimic the appearance of WSSV and highlight the importance of being able to fully characterise virus infections when they are first identified.
17
Fernandes, Ana Rita da Silva. "Terapêuticas das White Spot Lesions: revisão sistemática." Master's thesis, 2016. http://hdl.handle.net/10316/35402.
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Trabalho final do 5º ano com vista à atribuição do grau de mestre no âmbito do ciclo de estudos de Mestrado Integrado em Medicina Dentária apresentado à Faculdade de Medicina da Universidade de Coimbra.Introdução: A cárie dentária é uma das patologias mais comum e evitável. As primeiras manifestações da progressão da cárie dentária são denominadas por white spot lesions. Estas são definidas como uma desmineralização da superfície e subsuperfície do esmalte, sem que ocorra cavitação e com capacidade de serem revertidas. Objetivo: O objetivo desta revisão sistemática foi investigar quais os agentes de remineralização que são eficazes para o tratamento das white spot lesions. Materiais e Métodos: Para a realização desta revisão sistemática foi efetuada uma pesquisa bibliográfica nas bases de dados Pubmed, Cochrane Library e ScienceDirect com as seguintes palavras-chave: “white spot”, "tooth demineralization", "tooth remineralization", "fluorides”, conjugadas com os conectores boleanos “AND” e “OR”. Os critérios de inclusão foram: estudos clínicos, meta-análises, revisões de literatura e sistemáticas, de setembro de 2005 a setembro de 2015, redigidos em português ou inglês, com resumo disponível. Resultados: Da pesquisa inicial resultaram 273 referências. Após a eliminação de artigos duplicados resultaram 236 referências. Foram excluídas todas as referências não relevantes para a revisão sistemática, resultando 45 estudos potencialmente relevantes. Após leitura do texto integral resultou um total de 13 referências. Conclusão: São necessários mais estudos de evidência científica de modo a preconizar o método terapêutico mais adequado para o tratamento da desmineralização da superfície e subsuperfície do esmalte. Introduction: Dental caries are one of the most common and preventable diseases. The first sign of dental caries are denoted by white spot lesions (WSLs) which can be defined as a demineralization of the enamel surface and subsurface although these lesions can be reversed and do not form cavities. Objectives: The aim of this systematic review was investigate which remineralization agents are effective for the treatment of WSLs. Materials and Methods: For this systematic review a literature search was conducted on Pubmed, Cochrane Library and ScienceDirect with the following keywords: “white spot”, “tooth demineralization”, “tooth remineralization”, “fluorides”, combined with Boolean operators “AND” and “OR”. The inclusion criteria were: clinical trials, meta-analyzes, systematic and literature reviews, from September 2005 to September 2015, written in Portuguese or English, with available abstract. Results: The initial search resulted in 273 references. After elimination of duplicate articles resulted 236 references. After reading titles and abstracts, all non-relevant results were excluded, resulting in 45 potentially relevant studies. After reading the full text, 13 references were included. Conclusion: More studies are required for scientific evidence in order to reach a conclusion of the most suitable therapeutic method for the treatment of surface and subsurface demineralization of the enamel.
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Lin, An-Ting, and 林安婷. "Identification of white spot syndrome virus latency-associated genes." Thesis, 2004. http://ndltd.ncl.edu.tw/handle/37165342893905245543.
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碩士國立臺灣大學
動物學研究所
92
For recent years, the detection of white spot syndrome virus (WSSV) of general cultured black tiger shrimps in Taiwan reveals that as high as 90% shrimps are WSSV carriers. Latent viral infection generates a long-term relationship between virus and its host. Under the latent state, few viral genes are expressed, which establishes the latency stage. The study examines the transcriptional expression of WSSV 532 predictive open reading frames and further identifies the WSSV latent-associated genes by the reverse transcriptase- DNA polymerase chain reaction. It was found that wssv194, wssv313 and wssv369 express in the WSSV carried Penaues monodom, and the expression level increases with the infection time. The protein sequence analysis reveals that some predictive functional motifs of gene regulatory protein exist in the WSSV latency-associated genes. These three latency-associated genes might possibly be involved in the transcriptional regulation of WSSV latent stage.
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Huang, Ming-Hung, and 黃明宏. "Expression of three white spot syndrome virus (WSSV) genes." Thesis, 2005. http://ndltd.ncl.edu.tw/handle/23684908455959838090.
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碩士嘉南藥理科技大學
生物科技系暨研究所
93
White Spot Syndrome Virus (WSSV) is a large, circular dsDNA virus , which may infect shrimps and other crustaceans. This virus is a member of the genus Whispovirus within a new virus family called Nimaviridae, referred to the thread-like polar extension on the virus particle. The complete sequences of the WSSV genomes have now been published as three different isolates. However, most of WSSV open reading frames , which can encode proteins, have no homology to any known proteins or motifs. With in the genome of the Taiwan WSSV isolate, RT-PCR and microarry analyses showed that these ORFs (WSSV071, WSSV302,WSSV338) were putative as very late genes. Therefore, in this study, we try to construct the recombinant plasmids containing the ORFs, and to express them by transformation into Escherichia coli, BL-21(DE3). Until now, the His-tagged fusion proteins containing the WSSV071, WSSV302 and WSSV338 respectively were expressed successfully and purified to produce antiserums for Western blot analysis. In addition, we construct the recombinant plasmids containing the ORFs, and to express them by transfection into Spodoptera frugiperda (Sf9) insect cells .The efficiency of transfection were detected with EGFP expression using the fluorescent microscopy.
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Lin, Shinn-Tsuen, and 林信村. "Enzymatic Activity of Shrimp White Spot Syndrome Virus Ribonucleotide Reductase." Thesis, 2004. http://ndltd.ncl.edu.tw/handle/94565888632831263713.
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博士國立臺灣大學
動物學研究所
92
Infection of shrimps with white spot syndrome virus (WSSV) characterizes in fast virus replication. Therefore, the viral enzymes involved in DNA synthesis and metabolism must be very important. Among these enzymes, WSSV-encoded ribonucleotide reductase (RR) catalyzes the conversion of ribonucleoside diphosphates (NDP) to the corresponding deoxyribonucleoside diphosphates (dNDP), and thus plays a major role in DNA synthesis regulation. Moreover, previous studies show that the mRNA expression of both RR subunits increases concomitantly with the advancement of WSSV infection. A baculovirus/insect system was used to express the two recombinant subunits, RR1 and RR2. The SDS-PAGE electrophoresis showed high expression level of both recombinant subunits. The co-immunoprecipitation assay demonstrated that recombinant WSSV-RR1 and —RR2 reassembled in vitro. The DNA polymerase coupled RR activity assay showed a marked increase in RR activity when cell extracts containing recombinant RR1 and RR2, which are both required for activity, were combined. These experiment results demonstrated that recombinant WSSV-RR1 and —RR2 reassembled in vitro and formed active holoenzyme to catalyze the reaction. We found that EDTA and hydroxyurea inhibited rWSSV-RR activity. This suggests that metal ions and free radicals are required for its activity which is similar to the cases on eukaryotic cellular RRs. The reduction of cytidine 5’-diphophate (CDP) catalyzed by the recombinant WSSV RR was accelerated by ATP, a general activator of RR, in an optimal range of concentration. However, WSSV RR is different from the insect cellular RRs by its insensitivity to allosteric inhibition by dTTP and dATP. It is suggested that the tolerance of rWSSV-RR and cellular RR in feedback inhibition is different. RR activity increased in the gills and heart of experimentally infected shrimp as infection advanced. The increase is much higher in the gills than in the heart. Western blot analysis using specific polyclonal antibody raised against recombinant protein shows a concomitant increase in RR protein expression in WSSV-infected shrimp stomach tissues as the infection progresses. These results suggest that WSSV RR is functionally involved during WSSV infection.
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Wang, Han-Ching, and 王涵青. "Transcriptional analysis of four white spot syndrome virus (WSSV) genes." Thesis, 2002. http://ndltd.ncl.edu.tw/handle/06609840441061597814.
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碩士國立臺灣大學
海洋研究所
90
This study focused on a transcriptional analysis of four white spot syndrome virus genes ,vp28, vp22, WSSV262 and dnapol. Result from RT-PCR, which used for a temporal expression analysis, showed that WSSV262 and dnapol are early genes expressed 2 hrs post-infection, while vp28 and vp22 are late genes expressed 12 hrs post-infection. All four WSSV genes were monogenic and produced only one species of transcript, which suggests that these genes use their own specific promoters that are not shared with other genes. An analysis of the 5’ untranslated region (UTR) of WSSV262, dnapol and other early genes showed that the major transcriptional start points were located between 26th ~ 28th nucleotides downstream of the TATA box. Comparison of the 5’ UTR of these early genes revealed a consensus sequence of (a/t)CA(g/c/t)T(a/c),which is similar to the common initiator of arthropod genes and the mammalian TdT (terminal deoxynucleotidyltransferase) gene family. No consensus sequence was found from the 5’ UTR of WSSV late genes, vp28 and vp22. In addition to the conserved poly A signal in the 3’UTR of WSSV genes, two other consensus regions were identified around the poly A addition site of these genes: the poly A addition site is located one nucleotide downstream of a conserved TTT sequence in the AAATTTA element, while several nucleotides downstream of the poly A addition site, there is the conserved sequence TTTTATT, which is similar to the TTTTNT early transcriptional termination signal of poxvirus.
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Lai, ei-Chun, and 賴維川. "Development of the Detection Method for White Spot Baculovirus (WSBV)." Thesis, 1998. http://ndltd.ncl.edu.tw/handle/35770508503775724807.
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碩士國立中興大學
獸醫微生物學研究所
86
White spot baculovirus(WSBV) belongs to the genus Baculovirus of the family Baculoviridae. Currently, this virus is the most commonly found virus on shrimp farms and causes severe mortality in Asia and North America. In the past, diagnosis for WSBV was based on clinical signs and histopathological observation ; however, these methods such as the smear test, histopathological observation, or excrement test often result in the misjudgment in diagnosis. Recently, the advance of molecular biological techniques has offered another rapid method for diagnosis of illness related to shrimp disease with higher specificity and sensitivity. An accurate diagnosis and early prevention for WSBV should assist tominimize the mortality rate and reduce the economic losses. In this study, a Sal I restriction fragment approximately 4.6 kb in size of the WSBV genomic DNA was cloned and completely sequenced for nucleotide sequence analysis. Two primer pairs(3 and 4) were designed according to the nucleotide sequence of this particular DNA fragment and were subjected to polymerase chain reaction(PCR) experiments. PCR with primer pair 3 followed by a nested PCR with primer pair 4 could amplified a specific DNA fragment to WSBV genomic DNA approximately 2.3 kb in size. Furthermor, a DNA oligonucleotide probe specific to this WSBV genomic DNA fragment was labeled using DIG labeling system and used to detect the presence of WSBV DNA in the infected shrimp by dot blot hybridization or Southern blot hybridization. The results demonstrated that as little as 100 ng WSBV DNA could be detected by both methods confirmming the specificity and sensitivity of this probe. Additionally, development of serological reagents for WSBV is also important for diagnosis of this disease. Thus, the nucleotide sequence of this WSBV 4.6 kb Sal I restriction fragment was further analyzed by the computer GCG program and revealed six open reading frames : ORF 1 : 1~282, ORF 2 : 291~1493, ORF 3 : 2073~ 1750 (reverse), ORF 4 : 2360~2617, ORF 5 : 2971~3687, ORF 6: 3894~4284, respectively. The ORF 2 which encoded 400 amino acids was further cloned onto the expression vector pET28b(+) for production large amounts of this protein in E.coli. The expressed product of the recombinant plasmid pET28b(+)/pMS321B was a fusion protein with the molecular weight approximately 70 kDa which was identified by SDS-polyacrylamide gel electrophoresis and western blotting analysis. This expression product was further purified by His-bind affinity chromatography and then used as antigen to immunize mice for preparing specific antibody against WSBV.
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Chiu, Ya-Lin, and 邱雅琳. "The Infection of White Spot Syndrome Virus (WSSV) in Cultured." Thesis, 1997. http://ndltd.ncl.edu.tw/handle/97739995702253002678.
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碩士國立臺灣大學
動物學系
85
White spot syndrome virus (WSSV) is the causative agent of a disease which has been causing mass mortalities of cultured shrimps. The principle clinical sign of this disease is the presence of white spots on the exoskeleton, especially on the carapace of moribund shrimp. A Two-step WSSV diagnostic polymerase chain reaction (PCR), based upon the specific primer sets, was routinely used to detect WSSV in shrimps. Recently, white spots had also been observed on the 4th pleopod (ambulatory leg) of cultured crabs (Scylla serrata) and the carapace of wild-caught portunid crabs (Charybdis feriatus). When DNA was extracted from crabs collected from three sources: cultured crabs, wild-caught crabs and a non-cultured pest crab (Helice tridens), the PCR products showed the expected mobility, that is, they were coincident with the products amplified from the DNA prepared from WSSV-infected Penaeus monodon. The restriction profiles of these PCR products cleaved with HaeIII, HpaII, RsaI and Sau3AI were also the same. This is already strong evidence for the presence of WSSV in these various population. WSSV diagnostic PCR was also used to investigate the infection rate and the infected tissues of WSSV in wild- caught crabs. Almost all the tissues or organs of the seriously infected crabs showed WSSV positive after only one-step of amplification while tissues or organs of the lightly infected crabs were only positive in two-step WSSV PCR. The infection rate was particularly high in the gill, ambulatory leg and hemolymph, and only slightly less high in the stomach, eyestalk and maxilliped. The histopathological changes in the seriously infected tissues or organs were characterized by degenerated cells with hypertrophied nuclei. These nuclei were stained homogeneously by H & E. WSSV was confirmed as the causative agent by using in situ hybridization with a WSSV-specific probe. Various tissues from the mesoderm and ectoderm, such as connective tissue, epithelium, nervous tissue and muscle, could be infected by WSSV. Based on the number of the positive signals, the grades of infection could be distinguished. The gill, stomach and heart were the most seriously infected organs in crabs. Again, as with wild-caught shrimps which sometimes developed a patent infection after capture, the stress inherent in the cultured environment probably contributed to the wild- caught crabs﹐which were originally only lightly infected becoming seriously infected. In transmission electron micrographs of ultra-thin sections of infected tissues taken from infected wild-caught crabs (Portunus sanguinolentus), enveloped viral particles were readily observed in the nucleus of the infected cells. The viral agent was purified by CsCl gradient centrifugation. Negatively stained preparations showed the virus to be rod-shaped. The nucleocapsid measured 60-70 nm at its broadest point and was 325-350 nm long. The capsid was apparently composed of rings of subunits in a stacked series. The rings were aligned perpendicular to the longitudinal axis of the capsid. The thickness of the rings was very constant, usually being 20 nm. The morphological characteristics of this virus were thus very the similar to the viral agent purified from WSSV infected Penaeus monodon. Finally, healthy juvenile cultured crab, Scylla serrata, were exposed by immersion to epidermal filtrate from diseased P. monodon. Cumulative mortalities reached 40% within 2 weeks and WSSV was detected to be present in those experimentally infected crabs as early as 2 days post-infection. In situ hybridization with a WSSV-specific probe gave positive signals at 48 hr post- infection in the stomach, gill, cuticular epidermis and hepatopancreas. By 6 days post infection, almost all organs were heavily infected with WSSV. All of these results indicated that the causative agents of white spot syndrome of shrimps and crabs were in fact the same or closely related.
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Cheng-ShunCheng and 鄭丞舜. "Role of lipid metabolism in white spot syndrome virus replication." Thesis, 2016. http://ndltd.ncl.edu.tw/handle/bn268b.
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碩士國立成功大學
生物科技研究所
104
Global metabolic changes in WSSV-infected shrimp were recently clarified by us (using proteomics and metabolomics). In infected shrimp, re-routing host metabolism (analogous to the Warburg effect in cancer cells) increased availability of energy and building blocks in host cells at the genome replication stage (12 hours post infection; hpi). Moreover, WSSV switched lipid metabolism of host from lipolysis at 12 hpi to lipogenesis at 24 hpi and used it to complete the viral replication cycle and morphogenesis. At the replication stage (12 hpi), lipolysis induced by WSSV in hepatopancreas released free fatty acids that were rapidly assimilated by WSSV target tissues (e.g. hemocytes and stomach). Lipolysis switched to hemocytes and stomach until lipid in hepatopamcreas was exhausted at a late stage (24 hpi). Furthermore, beta-oxidation may be triggered during WSSV infection in shrimp. We determined that WSSV may trigger lipolysis in various tissues during viral replication, and that released free fatty acids may be absorbed by target tissues. Conversely, WSSV failed to complete its replication cycle after beta-oxidation was inhibited; this phenomenon was also observed following inhibition of fatty acid synthetase (FAS; a key enzyme of lipogenesis), during WSSV infection. Therefore, we inferred that alteration of lipid metabolism might be essential for WSSV virion formation. Our study provided new insights into important changes in host lipid metabolism triggered by an invertebrate virus.
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Lin, Chun-Wei, and 林君威. "Applicability Of Photodynamic Treatment To Inactivate White Spot Syndrome Virus." Thesis, 2014. http://ndltd.ncl.edu.tw/handle/04178525906923928178.
Full textAbstract:
碩士輔仁大學
生命科學系碩士班
102
White Spot Syndrome Virus (WSSV) is a serious viral disease to the shrimp culture industry. This study aims to create a powerful and energy-efficient method to inactive WSSV: Photodynamic treatment (PDT), to control the spread out of WSSV in shrimp’s culture pond. The WSSV was produced by Procambarus clarkii after muscle injection for 7 days .The energy-efficient LED and photosensitizing dyes were used in PDT for WSSV inactive. One hundred folds LD50 of WSSV were treated with PDT and injected into Litopenaeus vannamei’ muscle. The data showed the mortality rate were down from 100 to 33 % if the PDT is carry out with 160 mW /cm2 power consumed, 80 sec irradiation and 0.1 mM photosensitizing dyes. Excitingly, the mortality rate will down to zero only with the irradiate duration were increased to 100 sec. If the power consumed up to 225 mW /cm2, the mortality rate were down from 100 to 3 % with 20 sec irradiation. And the mortality rate also down to zero only if the irradiation time were up to 40 sec. Ten days after infection, the analysis of PCR for shrimp’s blood show that all of survival organisms don’t have any WSSV. On the other hands, if we take the healthy L. vannamei to carry out the PDT which could fully inactive WSSV (WPDT), results showed that the WPDT has not effect on the survival of shrimp during experiments. Under these results, we confirmed that the PDT has highly application potential to control the spread out of WSSV.
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Kroker, Tessa. "In- vitro- Untersuchung der Effektivität verschiedener Lacke zur Prävention von White- Spot- Läsionen im Rahmen kieferorthopädischer Behandlungen." Doctoral thesis, 2013. http://hdl.handle.net/11858/00-1735-0000-0015-9C00-4.
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Yen-I, Wu, and 吳彥儀. "Metabolic Effects of Infection of White Spot Syndrome Virus in the Pacific White Shrimp Penaeus vannamei." Thesis, 2014. http://ndltd.ncl.edu.tw/handle/88408059906972462832.
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碩士國立彰化師範大學
生物學系
102
The present study examined the effects of white spot syndrome virus (WSSV) on the Pev 20 system. Eyestalk ganglia Pev 20 levels were increased after WSSV infection, followed by a significant rise in hemolymph glucose levels; in addition, lactate levels in hemolymph did not change significantly after infection, but glucose levels in hemocyte increased significantly 24 hr after infection. On the other hand, glucose levels in muscle decreased significantly 36 hr after infection; glycogen levels in muscle did not change significantly after infection, whereas lactate levels in muscle increased significantly 12 hr after infection. With regard to glucose levels in hepatopancreas were significantly decreased 24 hr after infection, whereas glycogen levels in hepatopancreas did not change significantly after infection. Triglyceride levels in hepatopancreas were significantly decreased 6 hr, 12 hr after infection with 0 hr；glucose evels in gill and epidermis did not change significantly after infection； glycogen levels in gill and epidermis decreased 24 hr, 12 hr after infection, respectively. The combined results indicate that the Pev 20 systems are activated by WSSV, and downstream metabolite change. These WSSV-activated Pev 20 systems might be involved in response to pathogen-related stresses.
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Jyh-Ming, Tsai. "Investigations on the structural proteins of shrimp white spot syndrome virus." 2006. http://www.cetd.com.tw/ec/thesisdetail.aspx?etdun=U0001-2001200611134400.
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Lin, Yi-Hsing, and 林薏欣. "Expression analysis of white spot syndrome virus (WSSV) late gene p150." Thesis, 2002. http://ndltd.ncl.edu.tw/handle/87162558602676987185.
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碩士國立臺灣大學
動物學研究所
90
One of the white spot syndrome virus (WSSV) genes, p150, that encodes an acidic protein was identified and sequenced from the predicted open reading frame, WSSV067, from the Taiwan isolate. The p150 gene that contains 3,906 nucleotides potentially encodes a protein of 1,301 amino acids with two glutamic acid-rich domains in the N-terminus and three transmembrane regions in the C-terminus. The predicted molecular weight of P150 is 143.9 kDa. The transcript of p150, which was detected at 12 hr post-infection and reached a maximal level at 18 hr, belonged to a late gene of WSSV. The mRNA size of p150 gene is about 4.3 kb, and the poly A addition site is 15 bp downstream of the poly A signal. When compare the region between poly A signal and poly A addition site of known WSSV genes of p150, rr1, DNApol and the predicted open reading frame WSSV202 and WSSV217, a consensus sequence was found, (T/A)ACATATA(T/A)A(T/A). The product of in vitro transcription and translation of p150 gene was detected as a 150 kDa protein. Immunofluorescent staining of P150 protein in insect cell line showed that the P150 localized in cytoplasm, and might bind to some organelles. P150 did not associate with lysosome and mitochondria according to the observation under Confocal microscope, but might be colocalized with endoplasmic reticulum (ER) under fluorescence microscope. The P150 of WSSV may participate in virus assembly.
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Hsieh, Pei-Chein, and 謝佩倩. "The study of shrimp white spot syndrome virus - WSSV162 gene expression." Thesis, 2007. http://ndltd.ncl.edu.tw/handle/27916774048023930639.
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碩士嘉南藥理科技大學
生物科技系暨研究所
95
White spot syndrome virus (WSSV) is a widespread viral agent in shrimps and other crustaceans population. It has caused severe mortalities and huge enconomic losses to the shrimp farming industry, not only in Asia but also globally. Sequence analysis revealed a low level of homology between most WSSV ORFs and known genes from GeneBank, and this virus has been a member of the genus Whispovirus within a new virus family called Nimaviridae. This study is to investigate the function of one open reading frame (designated wssv162) from the WSSV genome of Taiwan isolate. We try to construct the recombinant plasmids containing the ORFs, and to express them by transformation into Escherichia coli, BL-21(DE3). Until now, the 6x His-tagged fusion proteins containing the WSSV162 were expressed successfully and used to prepare a specific antibody for western blotting. On the other hand, we transfected EGFP-WSSV162 construct into insect cells, and examined the cellular localization of the WSSV162 protein. The numbers of transfected insect cells, which expressed EGFP-WSSV162 fusion proteins, are low. But, it is interesting that EGFP-WSSV162 fusion proteins are mainly expressed in the cytoplasma of transfected insect cells. Then we also found that some EGFP-WSSV162 fusion proteins seem to be expressed as inclusion bodies. In order to study the interactions between WSSV162 protein and host shrimp proteins. We used the pull-down assay to analyze the possibilities of ineteractions between His6-WSSV162 fusion proteins from E. coli and shrimp proteins.
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Zhao, Zi-Lin, and 趙梓霖. "Characterization of White Spot Syndrome Virus (WSSV) Structural Protein VP38A (ORF314)." Thesis, 2010. http://ndltd.ncl.edu.tw/handle/70110167742224357379.
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碩士大葉大學
分子生物科技學系碩士班
98
White spot syndrome virus (WSSV) is a large DNA virus, which comprises three structural layers surrounding its core DNA, these layers include, an outer envelope, a tegument and a nucleocapsid. This study characterized a structural protein VP38A (WSSV-T1 ORF314, GenBank accession no. AF440570). Western blotting and immuno electronmicroscopy performed on the different salt concentrations Triton X-100 solution treated purified WSSV vireons, identified VP38A as an envelope protein. Membrane topology analysis showed that VP38A is exposed outside the virion but lacks a transmembrand domain. Co-immunoprecipitation assays demonstrated that VP38A could interact with VP28 and VP51A but not with VP19, VP24, VP26, VP32, VP37 and itself. Previous yeast two-hybrid screening experiment performed on a WSSV gene library displayed that VP38A was with autoactivation activity. This phenomenon was also confirmed there, implying that VP38A might act as a transcriptional factor. The above results demonstrate that VP38A is important, even essential, in WSSV infection and replication.
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Shiung, Hui-Jui, and 熊慧叡. "Characterization of White Spot Syndrome Virus (WSSV) Structural Protein VP11 (ORF394)." Thesis, 2010. http://ndltd.ncl.edu.tw/handle/70231360448356349845.
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碩士大葉大學
分子生物科技學系碩士班
98
White spot syndrome virus (WSSV) is a large enveloped virus. The WSSV viral particle consists of three structural layers that are surrounding its core DNA: an outer envelope, a tegument and a nucleocapsid. This study characterized a WSSV structural protein, VP11 (WSSV-T1 ORF394, GenBank accession no. AF440570). Immuno electromicroscopic analysis and Western blot hybridization of the intact viral particles and the separated viral components, the results showed that VP11 is an envelope protein. Membrane topology predition demonstrated that VP11 is a type transmembrane protein with a highly hydrophobic transmembrane domain at its N-terminal, and a C-terminal exposed on the virion surface. Immunofluorescence assay performed on VP11 transfected Sf9 cells demonstrated a similar result. However, trypsin digestion analysis of virions gave a controversial conclusion. Binary Co-immunopresipitation assays performed between VP11 and other a major WSSV structural proteins, respectively, VP11 was found to interact with both VP51A and itself. Yeast two-hybrid assay revealed that VP11 possessed autoactivation ability. We hypothesized that VP11 might act as a transcriptional factor. VP11 thus might play an important role in the replication of WSSV, not only in viral particle assembly but also in regulating at viral gene expression.
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Ye, YuJiun, and 葉鈺君. "Characterization of White Spot Syndrome Virus (WSSV) Structural Protein VP41B (ORF298)." Thesis, 2011. http://ndltd.ncl.edu.tw/handle/78941996078950323379.
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碩士大葉大學
分子生物科技學系碩士班
99
White spot syndrome virus (WSSV; genus Whispovirus, family Nimaviridae) is a widely occuring virus which attacks cultured shrimp and many other crustaceans and has caused severe mortalities and huge economic losses to the shrimp farming industry globally. WSSV is a large enveloped DNA virus. The protein components of the WSSV virion have been established and at least 58 structural proteins are currently known. In this study, a novel WSSV structural protein, VP41B (WSSV-TW ORF298) was characterized. vp41B is composed of 903 bp encoding a polypeptide of 300 aa with a theoretical mass of 41 kDa. Recombinant VP41B (r41B) was expressed and antibody against r41B was successfully produced. Western blot analysis of viral protein fractions suggested that VP41B is a viral envelope protein. Membrane topology assay demonstrated that the VP41B is located on the inner surface of envelope. Yeast two-hybrid assays revealed that VP41B associated directly with other 12 WSSV structural proteins and most of them are envelope proteins. VP41B is also with self-association activity forming several types of polymer. VP41B can also interact with 3 Penaeue monodon receptor proteins, including pmCBP, pmRACK1, and F1 ATP synthase beta subunitbeta subunit beta subunit. The above data suggest that the VP41B may be important in virion assembly and play a role in cell recognition, as well as in attaching the virus to the cell.
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Tsai, Jyh-Ming, and 蔡志明. "Investigations on the structural proteins of shrimp white spot syndrome virus." Thesis, 2006. http://ndltd.ncl.edu.tw/handle/26235175389852420555.
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博士國立臺灣大學
動物學研究研究所
94
White spot syndrome virus (WSSV) virions were purified from the hemolymph of experimentally infected crayfish Procambarus clarkii, and their proteins were separated by 8 to 18% gradient sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) to give a protein profile. The visible bands were then excised from the gel, and following trypsin digestion of the reduced and alkylated WSSV proteins in the bands, the peptide sequence of each fragment was determined by liquid chromatography–nano-electrospray ionization tandem mass spectrometry (LC-nanoESI-MS/MS) using a quadrupole/time-of-flight mass spectrometer. Comparison of the resulting peptide sequence data against the nonredundant database at the National Center for Biotechnology Information identified 33 WSSV structural genes, 20 of which are reported here for the first time. Since there were six other known WSSV structural proteins that could not be identified from the SDS-PAGE bands, there must therefore be a total of at least 39 (33 ＋ 6) WSSV structural protein genes. However, several details of the virus structure and assembly remain controversial, including the role of one of the major structural proteins, VP26. After isolating WSSV nucleocapsids by treatment with Triton X-100 and CsCl isopycnic equilibrium centrifugation, mass spectrometry identified only VP664 and four other minor structural proteins, VP160A, VP160B, VP60B, VP51C. Surprisingly, VP15 was not detected in this fraction. To locate the other structural proteins, 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, VP26, VP36A, VP39A, and VP95 were all identified as tegument proteins as 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 the above identified nucleocapsid component proteins using proteomic methods, by a trypsin sensitivity assay, and by an immunogold assay.
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Dixon, Julian Spencer. "Prevalence of white spot lesions during orthodontic treatment with fixed appliances /." 2009. http://hdl.handle.net/10156/2542.
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Ho, Cheng-Hung, and 何政鴻. "Construction of engineered bacterium that binds to white spot syndrome virus." Thesis, 2014. http://ndltd.ncl.edu.tw/handle/ktdh92.
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碩士國立高雄海洋科技大學
水產養殖研究所
102
White spot syndrome virus (WSSV) is a serious pathogen in shrimp culture, and there is no effective prevention method to resolve it so far. In order to reduce the concentration of WSSV in a breeding environment, this study aims to develop a bacterium that can bind with WSSV and remove it from water. Previous studies indicated that the WSSV Binding Protein (WBP) and Rab7 can bind to the VP26 and VP28 of WSSV, separately, in addition, the grouper Mx protein was also be demonstrated that Mx protein could bind with nervous necrosis virus (NNV) and against the infection of NNV. Therefore, we expressed WBP, Rab7 and Mx protein on bacterial surface to grenerate WSSV-binding bacterium and examine whether the bacterium can bind to the WSSV. The DNA fragments of WBP and Mx were amplified by assemble PCR, Rab7 gene was cloned by RT-PCR (Reverse Transcriptase PCR) and the RNA extracted from shrimp infected with WSSV.Subsequently, the virus binding genes were fused to C termini of lppompA or N-termini of AIDA, the Gram-negative surface display system, and formed lpp-WBP, lpp-Rab7, lpp-Mx, WBP-AIDA, Rab7-AIDA and Mx-AIDA, separatively. These recombinant plasmids were transformed into bacteria to generate lpp-WBP/BL21, lpp-Rab7/BL21, lpp-Mx/BL21, WBP-AIDA/BL21, Rab7-AIDA/BL21, and Mx-AIDA/BL21, separatively. Finally, we found that these bacteria can express protein on surface but failed to bind with WSSV by ELISA.
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Chun-YuanLi and 李春媛. "The role of glutaminolysis in White Spot Syndrome Virus (WSSV) pathogenesis." Thesis, 2015. http://ndltd.ncl.edu.tw/handle/5zq2cb.
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Tsai, Chia-Yi, and 蔡佳怡. "The study of shrimp white spot syndrome virus – wssv 044 gene expression." Thesis, 2008. http://ndltd.ncl.edu.tw/handle/89763811417482627534.
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碩士嘉南藥理科技大學
生物科技系暨研究所
96
White spot syndrome virus (WSSV), genus Whispovirus, family Nimaviridae, is a major shrimp pathogen that is highly virulent in penaeid shrimp and can also infect most species of crustacean. Because of the large size of WSSV genome and the uniqueness of viral proteins , only a few WSSV genes have been studied beyond sequence analysis. In this study, the ORF of wssv044 gene contained 201 nucleotides, encoding 67 a.a. The wssv044 gene was expressed in E. coli with His6-WSSV044 fusion protein ( 11-kDa ). The His6-WSSV044 were confirmed using Western bolt analysis with mouse anti-His antibodies. After 1mM IPTG and 37℃ induction for 3 hours, we found the His6-WSSV044 fusion proteins were express with inclusion body form. Subsequently, the expression of WSSV genes by fusing enhanced green fluorescent protein (EGFP) gene as a reporter gene were investigated in the insect Sf9 cells. First, we constructed and isolated the recombinant plasmids containing different WSSV genes respectively, which fused with N-terminal or C-terminal of EGFP. Then these recombinant plasmids were transfected into Sf9 cells and observed under a inverted fluorescence microscope. The results revealed that the fusion proteins containing different WSSV genes might be expressed diversely in transfected insect cells. A His pull-down assay has been used further to confirm the interactions between WSSV044 and shrimps proteins.
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Chou, Tsung-Lu, and 周宗錄. "Characterization of Shrimp White Spot Syndrome Virus (WSSV) Structural Protein VP51A (ORF294)." Thesis, 2007. http://ndltd.ncl.edu.tw/handle/68569959901509748570.
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碩士大葉大學
分子生物科技學系碩士班
95
White spot syndrome virus (WSSV) is an important crustacean virus causing high mortality in cultured shrimp. WSSV is a double-stranded DNA virus with a genome size of about 300 kbp. So far, 58 viral structural proteins were identified. In this research, one of the structural protein translated form ORF294 (GeneBank accession no. AF440570), the VP51A, was studied. Gene structure analysis showed that the transcription initiation site of vp51A was located 135 bp upstream of the translation start codon ATG. TATA box, or its related consensus sequence was not recognized is 5’ untranslated region of this gene. The poly-A addition signal was overlapped with the translation stop codon TAA and the poly-A tail was added 23 bp downstream of the stop codon.The vp51A transcripts was observed 6 hours after virus infection and the expression levels increasing with the infection time course. Computer software anlysis discovered a conserved sequence of the nuclear localization signal (NLS) between 37 and 43 of VP51A coding region, but such prediction wasn’t confirmed by the following in vitro analysis performed in Sf9 cells. Immunoelectron microscopy analysis and Western blot hybridization performed on intact virus particle and separated viral components showed that the VP51A is an envelope protein. Furthermore, Western blot analysis of WSSV virion also demonstrated that except the expected 53 kDa band, there were another protein bands such as an obvious signal around 72 kDa and some other small molecular weight proteins exist. Similar result was found in the Western blot results performed on WSSV infect shrimp tissues and recombinant VP51A expressed insect cells. But when using the in vitro transcription and translation system to express the recombinant VP51A it demonstrated a 72 kDa protein only. This result showed that the VP51A gene might expresses a large molecular weight protein first and it will then be processed into another lower molecular weight ones. Other experiments, including the predict protein cutting site mutation of VP51A and Western blot hybridization by VP51A different region fragments derived antibadies suggested that most of the cutting sites of VP51A might distribute closer to the N-terminal region. The is processed and what are the biological meanings of there different types of VP51A proteins, still left to be elocudate.
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Lin, Wei-Lun, and 林瑋倫. "Study on the Antiviral Mechanisms of Lactoferrin Against White Spot Syndrome Virus." Thesis, 2010. http://ndltd.ncl.edu.tw/handle/25707562105832332124.
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碩士大葉大學
分子生物科技學系碩士班
98
White spot syndrome virus (WSSV) is an important pathogen affecting cultured shrimp. To date these is no effective method to treat this disease caused by WSSV. Lactoferrin (Lf) is a multifunctional glycoprotein that is important in immune regulation and defense mechanisms against bacteria, fungi and viruses. Our previous research demonstrated that bovine Lf (bLf) treated shrimp are resistant to WSSV infection. In the present study, experiment performed on Autographa californica multiple nucleopolyhedrovirus (AcMNPV) and Sf9 insect cells and showed that bLf also interferes in AcMNPV infection. BLf treatment induced high level expression of certain immune-related genes in shrimp. Immonofluoresence assays demonstrated bLf was detected both in the nucleus and on the cell surface of bLf treated Sf9 cells and the hemocytes from bLf treated shrimp. Immunoelectromicroscopy, Far-Western blot and immunoprecipitation assays showed that the bLf was able to bind to the viral particle surface, and possibly via interaction with envelope proteins VP28. These results imply that the anti-viral effects of bLf on WSSV may be involved in multiple mechanisms, including upregulation of immune related gene expression, competing with WSSV to bind host cell surface molecules, and associating with viral particles to directly interfere in their the targeting on host cells.
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Chang, Po-Yuan, and 張博淵. "Interaction between White Spot Syndrome Virus Nonstructural Protein WSSV071 and Shrimp Transglutaminase." Thesis, 2013. http://ndltd.ncl.edu.tw/handle/45418792987974891483.
Full textAbstract:
碩士嘉南藥理科技大學
生物科技系
101
WSSV is a new large double-stranded DNA virus that infects aquatic crustaceans, and causes damages and losses to cultured shrimp. The around 300-kb genome of WSSV Taiwan isolate ( WSSV-TW ) encodes 532 putative open reading frames ( ORFs ). Some of them have been characterized encoding structural proteins of WSSV virions and many nonstructural proteins transiently expression during virus infection. It is study, time-course study using by real-time PCR showed that the wssv071 ORF of WSSV-TW was expressed after 24 hours of infection. The result suggested that wssv071 might be a late gene. Furthermore, Yeast-two hybrid analysis showed that WSSV071 protein might interact with the N-terminal and C-terminal peptide fragments of shrimp transglutaminase ( TGase ).
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Chen, Pei-Jung, and 陳姵蓉. "Identification and analysis of white spot syndrome virus (WSSV) envelope protein VP41A." Thesis, 2013. http://ndltd.ncl.edu.tw/handle/63158716935783347768.
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Yi, Der-Ming, and 易德明. "Transcript screening for very late genes of white spot syndrome virus(WSSV)." Thesis, 2004. http://ndltd.ncl.edu.tw/handle/35709161215136405381.
Full textAbstract:
碩士國立臺灣大學
動物學研究所
92
Microarray of time-course white spot syndrome virus (WSSV)-infected Penaeus monodon presents a transcription profile. The result reveals 13 open reading frames(orfs) that transcribed at approximately 36 hours post infection. Temporal analysis of 13 orfs by RT-PCR confirms the result of WSSV microarray and reveals that 8 orfs may be the very late gene candidates of WSSV. Motif and domain prediction results of WSSV very late gene candidates significantly discovered three types of regions that may contribute to the phenomena of WSSV infection very late stage. WSSV very late gene may function as anti-apoptosis genes or host immune evasions, which were advantageous in virion replication. Discovery of WSSV very late gene candidates provide a novel field of WSSV genomic research. In addition, functional analysis of WSSV very late gene candidates gives more implications about biological interaction between WSSV and its hosts. WSSV very late gene candidates may be crucial factors for providing solutions for the epidemic of WSSV.
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Liu, Wang-Jing, and 劉宛菁. "Studies on shrimp white spot syndrome virus(WSSV) immediate-early gene ie1." Thesis, 2007. http://ndltd.ncl.edu.tw/handle/36411224214247084722.
Full textAbstract:
博士國立臺灣大學
動物學研究研究所
95
Here we report for the first time the successful use of cycloheximide (CHX) as an inhibitor to block de novo viral protein synthesis during white spot syndrome virus (WSSV) infection. Sixty candidate immediate early (IE) genes were identified using a global analysis microarray technique. RT-PCR showed that the genes corresponding to ORF126, ORF242 and ORF418 in the Taiwan isolate were consistently CHX-insensitive, and these genes were designated ie1, ie2 and ie3, respectively. The sequences for these IE genes also appear in the two other WSSV isolates that have been sequenced. Three corresponding ORFs were identified in the China WSSV isolate, but only an ORF corresponding to ie1 was predicted in the Thailand isolate. In a promoter activity assay in Sf9 insect cells using enhanced green fluorescence protein (EGFP) as a reporter, ie1 showed very strong promoter activity, producing higher EGFP signals than the insect Orgyia pseudotsugata multicapsid nucleopolyhedrovirus ie2 promoter. Although the Janus kinase-signal transducer and activator of transcription (JAK-STAT) signaling pathway is part of the anti-viral response in arthropods such as Drosophila, here we show that WSSV uses shrimp STAT as a transcription factor to enhance viral gene expression in host cells. In a series of deletion and mutation assays using the WSSV immediate early gene ie1 promoter, which is active in shrimp cells and also in insect Sf9 cells, an element containing a STAT binding motif was shown to be important for the overall level of WSSV ie1 promoter activity. In the Sf9 insect cell line, a specific protein-DNA complex was detected by using electrophoresis mobility shift assays (EMSA) with the 32P-labeled STAT binding motif of the WSSV ie1 promoter as the probe. When recombinant Penaeus monodon STAT (rPmSTAT) was overexpressed in Sf9 cells, EMSA with specific antibodies confirmed that the STAT was responsible for the formation of the specific protein-DNA complex. Another EMSA showed that in WSSV-infected P. monodon, levels of activated PmSTAT were higher than in WSSV-free P. monodon. A transactivation assay of the WSSV ie1 promoter demonstrated that increasing the level of rPmSTAT led to dose-dependent increases in ie1 promoter activity. These results show that STAT directly transactivates WSSV ie1 gene expression and contributes to its high promoter activity. We conclude that WSSV has successfully annexed a putative shrimp defense mechanism, which it now uses to enhance the expression of viral immediate early genes.
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Liu, Wang-Jing, and 劉宛菁. "Study of the protein kinase gene of shrimp white spot syndrome virus." Thesis, 2000. http://ndltd.ncl.edu.tw/handle/32612336996494562116.
Full textAbstract:
碩士國立臺灣大學
動物學研究所
88
In the present study, a 3.4 kbp DNA fragment from white spot syndrome virus (WSSV) genomic HindIII library was studied. Following DNA sequencing and computer program prediction, five open reading frames (ORFs) were identified in this fragment. ORF1 has a high similarity to the protein kinase (PK) catalytic domain. ORF2 is identical to a previously reported WSSV envelope protein (VP28). Orf3, orf4 and orf5 potential encode for polypeptides of 193, 200 and 229 amino acids. Temporal analysis of the ORFs showed that ORF1, VP28 and ORF3 were first expressed at an early stage, but ORF4 and ORF5 were expressed at late stage. PK plays an important role in the pathogenicity of viruses, so this study focused on the cloning, sequencing, and characterization of WSSV pk1 gene. Northern blot analysis with an orf1-specific riboprobe detected a major transcript of 2.7 kb, which indicated that the 3.4 kbp fragment contained only part of WSSV pk1 gene. The full length WSSV pk1 gene was found in an 8421bp fragment from an EcoRI library. It contains 2193 nucleotides and encodes an 81-kDa protein of 730 amino acids. WSSV PK1 contains serine-threonine protein kinase consensus motifs I through XI. Compared to the PKs of other species, amino acid similarities are 24~41% for subdomains I and II; 69~92% for subdomain VI; 44~77% for subdomain VII; 45~63% for subdomain VIII; and 21~51% for subdomain IX. Analysis of WSSV pk1 gene structure indicated two TATA boxes at positions 75~80 and 14~19 nucleotides upstream of the translation start site, and a poly A signal (AATAAA) also located at the 3’ terminal of the gene. Expression of WSSV PK1 with a rabbit reticulocyte generated an 87-kDa protein, which is in close agreement with the predicted size of 84-kDa from the amino acids sequence. This in vitro transcription and translation product was recognized by anti-WSSV PK1 recombinant protein antiserum.
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Maxfield, Blake J. "Perceived responsibility for the development of white spot lesions during orthodontic treatment /." 2009. http://hdl.handle.net/10156/2541.
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Lin, Ting-An, and 林庭安. "Characterization of an immediate-early protein WSSV108 of white spot syndrome virus." Thesis, 2017. http://ndltd.ncl.edu.tw/handle/q2aesq.
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Gomes, Maria Luísa Lima da Quinta. "Agentes remineralizantes fluoretados e não fluoretados na abordagem das White Spot Lesions." Master's thesis, 2020. http://hdl.handle.net/10284/9355.
Full textAbstract:
Objetivo: Rever a bibliografia científica disponível sobre a abordagem preventivoterapêutica mais atual das White Spot Lesions (WSL) em pacientes pediátricos
recorrendo a agentes remineralizantes fluoretados e não fluoretados.
Metodologia: Pesquisa bibliográfica de artigos científicos publicados nas bases de
dados: PubMed, B-On e Web Of Science, entre 1999 e 2019. Foram articulados termos
de pesquisa através dos marcadores booleanos AND e NOT e definidos critérios de
inclusão e exclusão para a seleção dos manuscritos. No total 17 artigos foram
analisados.
Tópico abordado: As WSL representam um tipo de lesão bastante prevalente nas
crianças. O sucesso na abordagem preventivo-terapêutica destas lesões depende do
diagnóstico precoce e da aplicação de agentes remineralizantes, cujo objetivo deve
residir na prevenção e interceção destas lesões em detrimento de tratamentos mais
invasivos. Os agentes remineralizantes atuais proporcionam não só uma intervenção
eficaz com o aumento do potencial de remineralização das lesões, mas também
contribuem para uma melhoria da estética associada.Objective: Review the available scientific literature on the most current preventivetherapeutic approach to White Spot Lesions (WSL) in pediatric patients using fluoridated and non-fluoridated remineralizing agents. Methods: Bibliographic search of scientific articles published in the databases: PubMed, B-On and Web Of Science, between 1999 and 2019. Search terms were articulated using the Boolean AND and NOT markers and defined inclusion and exclusion criteria for the selection of manuscripts. In total, 17 articles were analyzed. Subject: WSL represents a type of injury that is quite prevalent in children. The success in the preventive-therapeutic approach of these lesions depends on the early diagnosis and the application of remineralizing agents, whose objective should be to prevent and intercept these lesions to the detriment of more invasive treatments. The current remineralizing agents provide not only an effective intervention with increasing the potential for remineralization of the lesions, but also contribute to an improvement of the associated aesthetics.
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Lin, Si-Wei, and 林思維. "Developing a Probiotics-Based Oral Vaccine for White Spot Syndrome in Prawns." Thesis, 2014. http://ndltd.ncl.edu.tw/handle/69700360507028417929.
Full textAbstract:
碩士國立高雄海洋科技大學
海洋生物技術研究所
102
Taiwan, a country renowned for its world-leading production of tiger prawns, has unfortunately been struck with white spot syndrome (WSS), severely reducing production. As the disease is currently incurable, developing a vaccine has become a top priority. This research sought to use probiotics to transport the envelope protein VP28 of the white spot syndrome virus (WSSV) as a type of vaccine. First, the characteristics of Pseudomonas sp. were analyzed, and the results showed that a salinity level of 0%~4% is suitable for the growth of Pseudomonas sp. and that its nitrite degradation ability is better than that of Bacillus sp. VP28 gene was cloned into pBBR1MCS-2 which does not require IPTG to induce, and then used electroporation to transform pMCS-VP28 into Pseudomonas sp. Two days after administering the VP28 vaccine to tiger prawns, VP28 was found in their intestines, and it was also found in their muscles after 4 days. A WSSV virus diluted to 5x105 was then used in the challenge tests. The 100ppm and 200ppm vaccine groups exhibited additional survival rates of 10% and 20%, respectively, compared to the control group. According to the immune parameter analysis, the haemocyte count, prophenoloxidase (proPO) activity, and phagocytosis ability of the vaccinated groups were all higher than those of the control group. This experiment verified that using the VP28 vaccine can improve the survival rate as well as the immunity of tiger prawns. After altering its medium, low-cost mass production of the vaccine can commence, allowing for widespread application in the future. Furthermore, it is hoped that by using Sia10, the vaccine can be converted into a secretable form for which additional field testing will be conducted.
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Saldanha, Thaylise Andrade. "Abordagem terapêutica das lesões de white spots associadas ao tratamento ortodôntico." Master's thesis, 2020. http://hdl.handle.net/10400.26/33979.
Full textAbstract:
Dissertação para obtenção do grau de Mestre no Instituto Universitário Egas MonizA incidência de cárie é observada em pacientes que se submetem a tratamento ortodôntico constitui uma preocupação em saúde oral, apesar deste tratamento proporcionar melhorias estéticas e funcionais. A aparatologia ortodôntica aumenta a probabilidade de desenvolvimento de lesões de white spot (WSLs), pois esses dispositivos favorecem a retenção de placa bacteriana e dificultam a higienização adequada das superfícies dentárias. O objetivo deste trabalho é elaborar uma revisão narrativa de forma a sintetizar e evidenciar a relação existente entre o tratamento ortodôntico e o aparecimento das lesões de white spot, assim como, abordar os métodos terapêuticos preconizados na literatura. Foi realizada uma pesquisa bibliográfica entre os anos de 1989 a 2020, através dos motores de busca científicos MEDLINE©/Pubmed©, Scielo©, ScienceDirect©, Lilacs© e plataforma B-On©. De acordo com a literatura publicada, pode-se concluir que entre os métodos utilizados para a terapêutica das WSLs, o uso de agentes remineralizantes e as resinas infiltrativas apresentam os melhores resultados no controlo dessas lesões.
The incidence of caries is observed in patients who undergo in orthodontic treatment is a concern in oral health, despite this treatment providing aesthetic and functional improvements. Orthodontic aparatology increases the probability of developing white spot lesions (WSLs), as these devices favor the retention of plaque and complicate the proper hygiene dental surfaces. The objective of this work is to elaborate a literature review in order to synthesize and highlight the relationship between orthodontic treatment and the appearance of white spot lesions, as well as addressing the therapeutic methods recommended in the literature. The bibliographic research was carried out between 1989 and 2020, using the scientific research engines MEDLINE © / Pubmed ©, Scielo ©, ScienceDirect ©, Lilacs © and the B-On © platform. According to the published literature, it can be concluded that among the methods used for the treatment of WSLs, the use of remineralizing agents and infiltrative resins show the best results in the control of these injuries.