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White spot syndrome virus: an overview on an emergent concern  

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Viruses are ubiquitous and extremely abundant in the marine environment. One of such marine viruses, the white spot syndrome virus (WSSV), has emerged globally as one of the most prevalent, widespread and lethal for shrimp populations. However, at present there is no treatment available to interfere...

Sánchez-Paz, Arturo

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Genetic variation among isolates of White Spot Syndrome Virus  

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

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

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The white spot syndrome virus DNA genome sequence  

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White spot syndrome virus (WSSV) is at present a major scourge to worldwide shrimp cultivation. We have determined the entire sequence of the double-stranded, circular DNA genome of WSSV, which contains 292,967 nucleotides encompassing 184 major open reading frames (ORFs). Only 6 f the WSSV ORFs hav...

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

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Prawn fry production process free from white spot syndrome virus  

UK PubMed Central (United Kingdom)

It relates to a technology of producing prawn seeds without white spotting syndrome virus. It comprises, first Selecting Parent prawn and temporary cultivated, then disinfecting the incubation pool with the bleaching powder, the potassium permanganate or the formaldehyde treating seawater for cultivating seeds with 5ppm bleaching powder (available chlorine content) and neutralizing detoxicatingova and incubating replacing worms with fish meals and steamed egg custard. The technology is nontoxic.

ZHANG QINGWEN; KONG JIE; LIU LONGCHANG

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

Directory of Open Access Journals (Sweden)

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

Desrina; Sarjito; Alfabetian Harjuno Condro Haditomo; Diana Chilmawati

2012-01-01

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White Spot Syndrome Virus Orf514 Encodes a Bona Fide DNA Polymerase  

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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...

Enrique de-la-Re-Vega; Karina D. Garcia-Orozco; Aldo A. Arvizu-Flores; Gloria Yepiz-Plascencia; Adriana Muhlia-Almazan

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

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Problem statement: White Spot Syndrome Virus (WSSV) is one of the most disastrous pathogens in shrimp culture, having caused high mortality in many cultured shrimp species. It is well known that the most important step of viral infection is cell attachment....

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

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

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White spot syndrome virus (WSSV) is a large DNA virus infecting shrimp and other crustaceans. The virus particles contain at least five major virion proteins, of which three (VP26, VP24, and VP15) are present in the rod-shaped nucleocapsid and two (VP28 and VP19) reside in the envelope. The mode of ...

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

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

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

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

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Protection of Penaeus monodon against White Spot Syndrome Virus by oral vaccination  

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White spot syndrome virus (WSSV) occurs worldwide and causes high mortality and considerable economic damage to the shrimp farming industry. No adequate treatments against this virus are available. It is generally accepted that invertebrates such as shrimp do not have an adaptive immune response sys...

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

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

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White spot syndrome virus (WSSV) is a virus infecting shrimp and other crustaceans, which is unclassified taxonomically. A 2193 bp long open reading frame, encoding a putative protein kinase (PK), was found on a 8.4 kb EcoRI fragment of WSSV proximal to the gene for the major envelope protein (VP28)...

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

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

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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 st...

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

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Protection of Penaeus monodon against White Spot Syndrome Virus by Oral Vaccination  

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White spot syndrome virus (WSSV) occurs worldwide and causes high mortality and considerable economic damage to the shrimp farming industry. No adequate treatments against this virus are available. It is generally accepted that invertebrates such as shrimp do not have an adaptive immune response sys...

Witteveldt, Jeroen; Cifuentes, Carolina C.; Vlak, Just M.; van Hulten, Mariëlle C. W.

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

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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 ...

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

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Virion composition and genomics of white spot syndrome virus of shrimp  

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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...

Hulten, M.C.W., van

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

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White spot syndrome is a worldwide disease of penaeid shrimp. The disease agent is a bacilliform, enveloped virus, white spot syndrome virus (WSSV), with a double-stranded DNA genome that probably contains well over 200 kb. Analysis of a 12?3 kb segment of WSSV DNA revealed eight open reading frames...

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

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

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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 ...

Sutee WONGMANEEPRATEEP; Puttharat BAOPRASERTKUL; Piyanuch PROMPAMORN; Kanittada THONGKAO; Chalor LIMSUWAN; Niti CHUCHIRD

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Increased susceptibility of white spot syndrome virus-infected Litopenaeus vannamei to Vibrio campbellii  

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The concept of polymicrobial disease is well accepted in human and veterinary medicine but has received very little attention in the field of aquaculture. This study was conducted to investigate the synergistic effect of white spot syndrome virus (WSSV) and Vibrio campbellii on development of...

Phuoc, L.H.; Corteel, M.; Nauwynck, H.J.; Pensaert, M.B.; Alday-Sanz, V.; Van Den Broeck, W.; Sorgeloos, P.; Bossier, P.

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

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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), cr...

González-Galaviz José Reyes; Rodriguez-Anaya Libia Zulema; Molina-Garza Zinnia Judith; Ibarra-Gámez José Cuauhtémoc

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

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Early events in white spot syndrome virus (WSSV) morphogenesis, particularly the formation of viral membranes, are poorly understood. The major envelope proteins of WSSV are VP28, VP26, VP24, and VP19. Our previous results indicated that VP28 interacts with VP26 and VP24. In the present study, we us...

Zhou, Qing; Xu, Limei; Li, Hui; Qi, Yi-Peng; Yang, Feng

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

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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 v...

Wu, Chenglin; Yang, Feng

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

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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 ...

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

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

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Abstract Transmission of white spot syndrome virus (WSSV) in shrimp has been reported to occur by feeding and immersion. In the present study, the impact of the molt process and artificial lesions in the cuticle on shrimp susceptibility to WSSV was examined using intramuscular and immersion ...

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

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White spot syndrome virus (WSSV) virions were purified from the tissues of infected Procambarus clarkii (crayfish) isolates. Pure WSSV preparations were subjected to Triton X-100 treatment to separate into the envelope and nucleocapsid fractions, which were subsequently separated by 12% sodium dodec...

Xie, Xixian; Xu, Limei; Yang, Feng

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

Science.gov (United States)

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.

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

2000-09-01

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

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White spot syndrome virus (WSSV) is one major pathogen in shrimp aquaculture. WSSV ORF403 is predicted to encode a protein of 641 amino acids, which contains a C3H2C2 RING structure. In the presence of an E2 conjugating enzyme from shrimp, WSSV403 can ubiquitinate itself in vitro, indicating it can ...

He, Fang; Kwang, Jimmy

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

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In a study of genes expressed differentially in the freshwater crayfish Pacifastacus leniusculus infected experimentally with the white spot syndrome virus (WSSV), one protein, known as antilipopolysaccharide factor (ALF), was chosen, among those whose transcript levels increased upon viral infectio...

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

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

Directory of Open Access Journals (Sweden)

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

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

2009-01-01

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Analysis of the complete nucleotide sequence of a white spot syndrome virus isolated from pacific white shrimp.  

UK PubMed Central (United Kingdom)

The fourth complete genome sequence of the white spot syndrome virus (WSSV) from Pacific white shrimp (Litopenaeus vannamei) isolated from Korea (WSSV-KR) was determined. The genome is composed of 295,884 bp encompassing 515 open reading frames (ORFs) among which 90 showed no sequence homology with any known protein in BLAST searches. The remaining 425 ORFs encode functional proteins including enzymes for nucleic acid metabolism, DNA replication and transcription, and several major structural proteins. Dot plot and linear comparisons of WSSV Korean strain with other WSSV isolates showed overall similarity but with some areas of sequence difference and one large deletion area.

Chai CY; Yoon J; Lee YS; Kim YB; Choi TJ

2013-09-01

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Designing and Introduce a Diagnostic Kit for Detection of White Spot Syndrome Virus in Cultured Penaeus indicus in Iran  

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Full Text Available The aim of this study is designing a diagnostic kit for white spot syndrome virus. We designed 2 series of primers for diagnosis of viral VP24 gene and also primers as internal controls which amplify part of genome in both positive and negative samples. DNA of shrimps were extracted and PCR amplification carried out. In this research, a diagnosis kit for white spot disease of shrimps designed and tested using 32 shrimp samples which were dubious to have this disease. White spot virus were found in 23 samples and the other 9 were negative. For extra confirmation, the PCR product was sequenced and deposited to GenBank. We designed a diagnosis kit for white spot disease of shrimps and tested successfully.

A.M. Saberi; M. Bandehpour; M. Afsharnasab; E. Ghayour; S.A. Yousefi Namin; B. Kazemi

2008-01-01

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Genome dynamics in three different geographical isolates of white spot syndrome virus (WSSV).  

Science.gov (United States)

White spot syndrome virus (WSSV), the sole member of the monotypic family Nimaviridae, is considered an extremely lethal shrimp pathogen. Despite its impact, some essential biological characteristics related to WSSV genome dynamics, such as the synonymous codon usage pattern and selection pressure in genes, remain to be elucidated. The results show that compositional limitations and mutational pressure determine the codon usage bias and base composition in WSSV. Furthermore, different forces of selective pressure are acting across various regions of the WSSV genome. Finally, this study points out the possible occurrence of two major recombination events. PMID:22836599

Sablok, Gaurav; Sánchez-Paz, Arturo; Wu, Xianming; Ranjan, Jayant; Kuo, Jimmy; Bulla, Ingo

2012-07-27

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Genome dynamics in three different geographical isolates of white spot syndrome virus (WSSV).  

UK PubMed Central (United Kingdom)

White spot syndrome virus (WSSV), the sole member of the monotypic family Nimaviridae, is considered an extremely lethal shrimp pathogen. Despite its impact, some essential biological characteristics related to WSSV genome dynamics, such as the synonymous codon usage pattern and selection pressure in genes, remain to be elucidated. The results show that compositional limitations and mutational pressure determine the codon usage bias and base composition in WSSV. Furthermore, different forces of selective pressure are acting across various regions of the WSSV genome. Finally, this study points out the possible occurrence of two major recombination events.

Sablok G; Sánchez-Paz A; Wu X; Ranjan J; Kuo J; Bulla I

2012-12-01

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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  

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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 ...

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

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Prevalence of white spot syndrome virus infection detected by one-step and nested PCR in selected tiger shrimp (Penaeus monodon) hatcheries  

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White spot syndrome (WSS) is considered as a great threat to commercial farming of the tiger shrimp (Penaeus monodon). The causal agent of WSS is a DNA virus called white spot syndrome virus (WSSV). The prevalence of this dreadful virus infection has been studied in five randomly selected hatcheries...

Ayub, Farhana; Sarker, Md.; Alam, Md.

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

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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, VP...

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

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White spot syndrome virus protein ICP11: A histone-binding DNA mimic that disrupts nucleosome assembly  

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White spot syndrome virus (WSSV) is a large (?300 kbp), double-stranded DNA eukaryotic virus that has caused serious disease in crustaceans worldwide. ICP11 is the most highly expressed WSSV nonstructural gene/protein, which strongly suggests its importance in WSSV infection; but until now, its func...

Wang, Hao-Ching; Wang, Han-Ching; Ko, Tzu-Ping; Lee, Yu-May; Leu, Jiann-Horng; Ho, Chun-Han; Huang, Wei-Pang; Lo, Chu-Fang

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

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White Spot Syndrome Virus (WSSV) is an infectious pathogen of shrimp and other crustaceans, and neither effective vaccines nor adequate treatments are currently available. WSSV is an enveloped dsDNA virus, and one of its major envelope proteins, VP28, plays a pivotal role in WSSV infection. In an at...

S, Syed Musthaq; Kwang, Jimmy

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

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White spot syndrome virus (WSSV) is a taxonomically unclassified virus which causes a disease in shrimps worldwide. A 936 bp long open reading frame (ORF) was found on a 7.2 kb HindIII fragment of the DNA genome of WSSV located adjacent to the ribonucleotide reductase small subunit gene. This putati...

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

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White Spot Syndrome Virus Orf514 Encodes a Bona Fide DNA Polymerase  

Directory of Open Access Journals (Sweden)

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.

Enrique de-la-Re-Vega; Karina D. Garcia-Orozco; Aldo A. Arvizu-Flores; Gloria Yepiz-Plascencia; Adriana Muhlia-Almazan; Jesús Hernández; Luis G. Brieba; Rogerio R. Sotelo-Mundo

2011-01-01

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

UK PubMed Central (United Kingdom)

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, ?-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.

Ye T; Zong R; Zhang X

2012-08-01

 
 
 
 
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VP90 of white spot syndrome virus interacts with VP26 and VP28.  

UK PubMed Central (United Kingdom)

Identification of structural protein relationships is likely to be important for virus assembly and anti-WSSV strategies. In this paper, VP90 of white spot syndrome virus (WSSV) was characterized. Temporal-transcription analysis showed that VP90 is expressed in the late stage of WSSV infection. Far-western and matrix-assisted laser desorption ionization mass spectrometry (MALDI-MS) assay showed that VP90 interacts with VP26 and VP28, two major envelope proteins. Far-western blot with recombinant VP26 (rVP26), N-terminal end of VP28 (rVP28N) and C-terminal end of VP28 (rVP28C) provided further evidence for interaction of rVP90 and rVP26 or rVP28. These results suggest that VP90 is anchored to the envelope through interacting with VP26 and VP28.

Li Q; Liu QH; Huang J

2012-01-01

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

UK PubMed Central (United Kingdom)

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.

Shashikumar A; Desai PV

2013-03-01

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

Science.gov (United States)

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. PMID:22828752

Shashikumar, Anumol; Desai, P V

2012-07-25

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

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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.

Leu Jiann-Horng; Chang Chih-Chin; Wu Jin-Lu; Hsu Chun-Wei; Hirono Ikuo; Aoki Takashi; Juan Hsueh-Fen; Lo Chu-Fang; Kou Guang-Hsiung; Huang Hsuan-Cheng

2007-01-01

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Natural occurrence of White spot syndrome virus and Infectious hypodermal and hematopoietic necrosis virus in Neohelice granulata crab.  

Science.gov (United States)

White spot syndrome virus (WSSV) and Infectious hypodermal and hematopoietic necrosis virus (IHHNV) are two infectious agents associated to economic losses in shrimp aquaculture. As virus spread occurs through vectors and hosts, this study sought to verify the presence of WSSV and IHHNV in Neohelice granulata crab from Lagoa dos Patos estuary in Brazil and nearby shrimp farms. DNA extractions were performed with phenol/chloroform protocol. Molecular diagnosis was carried out by nested PCR for WSSV and one-step PCR for IHHNV. Results showed the presence of WSSV on crabs of both Lagoa dos Patos and farms, while IHHNV was found only on crabs collected in estuary. This is the first study to report IHHNV presence in N. granulata. Moreover, as analyzed crabs had no clinical symptoms or showed in situ mortality, we suggest its use as a bioindicator for virus occurrence in aquatic environments. PMID:23792092

Cavalli, Lissandra Souto; Batista, Carolina Reyes; Nornberg, Bruna F S; Mayer, Fabiana Quoos; Seixas, Fabiana K; Romano, Luis Alberto; Marins, Luis Fernando; Abreu, Paulo César

2013-06-17

46

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

Science.gov (United States)

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

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

2013-08-26

47

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

UK PubMed Central (United Kingdom)

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

Chen X; Zeng D; Chen X; Xie D; Zhao Y; Yang C; Li Y; Ma N; Li M; Yang Q; Liao Z; Wang H

2013-01-01

48

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

Science.gov (United States)

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

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

2013-01-01

49

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

UK PubMed Central (United Kingdom)

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.

Feng S; Li G; Feng W; Huang J

2013-10-01

50

Characterization of white spot syndrome virus immediate-early gene promoters.  

Science.gov (United States)

Twenty-one immediate-early (IE) genes of white spot syndrome virus (WSSV) have been identified so far. However, the transcriptional regulation of WSSV IE genes remains largely unknown. In this report, the 5' flanking regions of 18 WSSV IE genes were cloned and eight functional promoter regions were identified. WSSV IE gene promoters normally contained a TATA box approximately 30 bp upstream of the transcriptional initiation site. Also, the cyclic AMP response element (CRE; TGACGTCA) was frequently found within the WSSV IE promoter regions. Mutations of the CREs of WSSV IE promoters P403 and P465 reduced their activity significantly, suggesting that these elements have a role in WSSV IE gene transcription. Our findings provide a more global view of WSSV IE gene promoters and will facilitate the in-depth investigation of viral gene transcriptional regulation. PMID:23136364

Lin, Fanyu; Huang, He; Ke, Wei; Hou, Luhong; Li, Fang; Yang, Feng

2012-11-07

51

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

Directory of Open Access Journals (Sweden)

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.

González-Galaviz José Reyes; Rodriguez-Anaya Libia Zulema; Molina-Garza Zinnia Judith; Ibarra-Gámez José Cuauhtémoc; Galaviz-Silva Lucio

2013-01-01

52

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

Energy Technology Data Exchange (ETDEWEB)

The nonstructural protein VP9 from white spot syndrome virus (WSSV) has been identified and expressed in Escherichia coli. To facilitate purification, a cleavable His{sub 6} tag was introduced at the N-terminus. The native protein was purified and crystallized by vapor diffusion against mother liquor containing 2 M sodium acetate, 100 mM MES pH 6.3, 25 mM cadmium sulfate and 3% glycerol. Crystals were obtained within 7 d and diffracted to 2.2 Angstroms; they belonged to space group P2{sub 1}2{sub 1}2{sub 1}, with unit-cell parameters a = 74.13, b = 78.21, c = 78.98 Angstroms and four molecules in the asymmetric unit. The selenomethionine-labeled protein produced isomorphous crystals that diffracted to approximately 3.3 Angstroms.

Liu,Y.; Sivaraman, J.; Hew, C.

2006-01-01

53

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

International Nuclear Information System (INIS)

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

2005-07-05

54

Endocytic pathway is indicated for white spot syndrome virus (WSSV) entry in shrimp.  

Science.gov (United States)

The white spot syndrome virus (WSSV) has had a serious economic impact on the global shrimp aquaculture industry in the past two decades. Although research has clarified a lot about its genome and structure, the mechanism of how WSSV enters a cell is still unclear. In this study to determine this mechanism, primary cultured hemocytes were used as an experimental model to observe the process of WSSV entry because the stable shrimp cell lines for WSSV infection are lacking. After labeling virions and endosomes with fluorescent dyes followed by observation with a confocal microscope, the results show that the WSSV colocalizes with early endosomes. Hemocytes are further treated with different endocytic inhibitors, methyl-?-cyclodextrin (M?CD) and chlorpromazine (CPZ). WSSV still can be detected in the hemocytes treated with CPZ, but not in the hemocytes treated with M?CD. Thus, we conclude that WSSV adopts the caveolae-mediated endocytosis to enter the shrimp cell. PMID:23747417

Huang, Zih-Jhan; Kang, Shih-Ting; Leu, Jiann-Horng; Chen, Li-Li

2013-06-06

55

Characterization of white spot syndrome virus immediate-early gene promoters.  

UK PubMed Central (United Kingdom)

Twenty-one immediate-early (IE) genes of white spot syndrome virus (WSSV) have been identified so far. However, the transcriptional regulation of WSSV IE genes remains largely unknown. In this report, the 5' flanking regions of 18 WSSV IE genes were cloned and eight functional promoter regions were identified. WSSV IE gene promoters normally contained a TATA box approximately 30 bp upstream of the transcriptional initiation site. Also, the cyclic AMP response element (CRE; TGACGTCA) was frequently found within the WSSV IE promoter regions. Mutations of the CREs of WSSV IE promoters P403 and P465 reduced their activity significantly, suggesting that these elements have a role in WSSV IE gene transcription. Our findings provide a more global view of WSSV IE gene promoters and will facilitate the in-depth investigation of viral gene transcriptional regulation.

Lin F; Huang H; Ke W; Hou L; Li F; Yang F

2013-02-01

56

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

Directory of Open Access Journals (Sweden)

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.

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

2012-01-01

57

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

International Nuclear Information System (INIS)

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

2004-07-20

58

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

Science.gov (United States)

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. PMID:23711885

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

2013-05-24

59

Crassostrea gigas oysters as a shrimp farm bioindicator of white spot syndrome virus.  

UK PubMed Central (United Kingdom)

This study explored whether Crassostrea gigas oysters can be used as a bioindicator of white spot syndrome virus (WSSV) in shrimp farm water canals. Bioassays showed that C. gigas can accumulate WSSV in their gills and digestive glands but do not become infected, either by exposure to seawater containing WSSV or by cohabitation with infected shrimp. The use of a WSSV nested PCR to screen oysters placed in water canals at the entry of a shrimp farm allowed WSSV to be detected 16 d prior to the disease occurring. The finding that C. gigas can concentrate small amounts of WSSV present in seawater without being harmed makes it an ideal sentinel species at shrimp farms.

Vazquez-Boucard C; Escobedo-Fregoso C; Duran-Avelar Mde J; Mercier L; Llera-Herrera R; Escobedo-Bonilla C; Vibanco-Perez N

2012-04-01

60

Indian isolates of white spot syndrome virus exhibit variations in their pathogenicity and genomic tandem repeats.  

UK PubMed Central (United Kingdom)

To detect genomic variation of white spot syndrome virus (WSSV) isolates from different geographical regions of India, the variable number of the tandem repeat (VNTR) region of the ORF 94 (Thailand WSSV isolate - GeneBank Accession No. AF369029) was analysed using five specific sets of primers. Analysis of 70 WSSV-positive samples showed the presence of 14 different genotypes of WSSV with VNTRs ranging from 2 to 16 tandem repeats with the majority (85.47%) having 6-12 tandem repeats. Occurrence of different genotypes of WSSV was found to be neither correlated to any specific geographical region nor to the different growth stage of the tiger shrimp, Penaeus monodon. Pathogenicity studies conducted with 25 isolates of WSSV revealed the presence of virulent and avirulent strains of WSSV in Indian shrimp farms. However, an unambiguous link could not be established between the different genotypes and their virulence.

John KR; George MR; Iyappan T; Thangarani AJ; Jeyaseelan MJ

2010-09-01

 
 
 
 
61

Indian isolates of white spot syndrome virus exhibit variations in their pathogenicity and genomic tandem repeats.  

Science.gov (United States)

To detect genomic variation of white spot syndrome virus (WSSV) isolates from different geographical regions of India, the variable number of the tandem repeat (VNTR) region of the ORF 94 (Thailand WSSV isolate - GeneBank Accession No. AF369029) was analysed using five specific sets of primers. Analysis of 70 WSSV-positive samples showed the presence of 14 different genotypes of WSSV with VNTRs ranging from 2 to 16 tandem repeats with the majority (85.47%) having 6-12 tandem repeats. Occurrence of different genotypes of WSSV was found to be neither correlated to any specific geographical region nor to the different growth stage of the tiger shrimp, Penaeus monodon. Pathogenicity studies conducted with 25 isolates of WSSV revealed the presence of virulent and avirulent strains of WSSV in Indian shrimp farms. However, an unambiguous link could not be established between the different genotypes and their virulence. PMID:20690959

John, K R; George, M R; Iyappan, T; Thangarani, A J; Jeyaseelan, M J P

2010-09-01

62

A model for apoptotic interaction between white spot syndrome virus and shrimp.  

Science.gov (United States)

White spot syndrome virus (WSSV) is an enveloped, large dsDNA virus that mainly infects penaeid shrimp, causing serious damage to the shrimp aquaculture industry. Like other animal viruses, WSSV infection induces apoptosis. Although this occurs even in by-stander cells that are free of WSSV virions, apoptosis is generally regarded as a kind of antiviral immune response. To counter this response, WSSV has evolved several different strategies. From the presently available literature, we construct a model of how the host and virus both attempt to regulate apoptosis to their respective advantage. The basic sequence of events is as follows: first, when a WSSV infection occurs, cellular sensors detect the invading virus, and activate signaling pathways that lead to (1) the expression of pro-apoptosis proteins, including PmCasp (an effecter caspase), MjCaspase (an initiator caspase) and voltage-dependent anion channel (VDAC); and (2) mitochondrial changes, including the induction of mitochondrial membrane permeabilization and increased oxidative stress. These events initiate the apoptosis program. Meanwhile, WSSV begins to express its genes, including two anti-apoptosis proteins: AAP-1, which is a direct caspase inhibitor, and WSV222, which is an E3 ubiquitin ligase that blocks apoptosis through the ubiquitin-mediated degradation of shrimp TSL protein (an apoptosis inducer). WSSV also induces the expression of a shrimp anti-apoptosis protein, Pm-fortilin, which can act on Bax to inhibit mitochondria-triggered apoptosis. This is a life and death struggle because the virus needs to prevent apoptosis in order to replicate. If WSSV succeeds in replicating in sufficient numbers, this will result in the death of the infected penaeid shrimp host. PMID:22683516

Leu, Jiann-Horng; Lin, Shin-Jen; Huang, Jiun-Yan; Chen, Tsan-Chi; Lo, Chu-Fang

2012-06-07

63

A model for apoptotic interaction between white spot syndrome virus and shrimp.  

UK PubMed Central (United Kingdom)

White spot syndrome virus (WSSV) is an enveloped, large dsDNA virus that mainly infects penaeid shrimp, causing serious damage to the shrimp aquaculture industry. Like other animal viruses, WSSV infection induces apoptosis. Although this occurs even in by-stander cells that are free of WSSV virions, apoptosis is generally regarded as a kind of antiviral immune response. To counter this response, WSSV has evolved several different strategies. From the presently available literature, we construct a model of how the host and virus both attempt to regulate apoptosis to their respective advantage. The basic sequence of events is as follows: first, when a WSSV infection occurs, cellular sensors detect the invading virus, and activate signaling pathways that lead to (1) the expression of pro-apoptosis proteins, including PmCasp (an effecter caspase), MjCaspase (an initiator caspase) and voltage-dependent anion channel (VDAC); and (2) mitochondrial changes, including the induction of mitochondrial membrane permeabilization and increased oxidative stress. These events initiate the apoptosis program. Meanwhile, WSSV begins to express its genes, including two anti-apoptosis proteins: AAP-1, which is a direct caspase inhibitor, and WSV222, which is an E3 ubiquitin ligase that blocks apoptosis through the ubiquitin-mediated degradation of shrimp TSL protein (an apoptosis inducer). WSSV also induces the expression of a shrimp anti-apoptosis protein, Pm-fortilin, which can act on Bax to inhibit mitochondria-triggered apoptosis. This is a life and death struggle because the virus needs to prevent apoptosis in order to replicate. If WSSV succeeds in replicating in sufficient numbers, this will result in the death of the infected penaeid shrimp host.

Leu JH; Lin SJ; Huang JY; Chen TC; Lo CF

2013-04-01

64

Immune response of white shrimp, Litopenaeus vannamei, after a concurrent infection with white spot syndrome virus and infectious hypodermal and hematopoietic necrosis virus.  

Science.gov (United States)

In the present study, we investigated immunological changes in viral-infected white shrimp, Litopenaeus vannamei. White shrimp were infected with white spot syndrome virus (WSSV) or co-infected with WSSV and infectious hypodermal and hematopoietic necrosis virus (IHHNV) as detected by polymerase chain reaction (PCR). The complete (100%) mortality rate of shrimp was caused by viral infection due to immune parameters being suppressed including decreases in phenoloxidase activity, total hemocyte counts, differential hemocyte counts, and the gene expressions of prophenoloxidase and peroxinectin. In addition, increases in lipopolysaccharide and beta-1,3-glucan-binding protein of hemocytes and the hepatopancreas, and respiratory bursts per cell, and a decrease in superoxide dismutase were found in viral-infected shrimp, which may have been related to the defense against viral infection. PMID:18948207

Yeh, Shinn-Pyng; Chen, Ying-Nan; Hsieh, Shu-Ling; Cheng, Winton; Liu, Chun-Hung

2008-10-01

65

Immune response of white shrimp, Litopenaeus vannamei, after a concurrent infection with white spot syndrome virus and infectious hypodermal and hematopoietic necrosis virus.  

UK PubMed Central (United Kingdom)

In the present study, we investigated immunological changes in viral-infected white shrimp, Litopenaeus vannamei. White shrimp were infected with white spot syndrome virus (WSSV) or co-infected with WSSV and infectious hypodermal and hematopoietic necrosis virus (IHHNV) as detected by polymerase chain reaction (PCR). The complete (100%) mortality rate of shrimp was caused by viral infection due to immune parameters being suppressed including decreases in phenoloxidase activity, total hemocyte counts, differential hemocyte counts, and the gene expressions of prophenoloxidase and peroxinectin. In addition, increases in lipopolysaccharide and beta-1,3-glucan-binding protein of hemocytes and the hepatopancreas, and respiratory bursts per cell, and a decrease in superoxide dismutase were found in viral-infected shrimp, which may have been related to the defense against viral infection.

Yeh SP; Chen YN; Hsieh SL; Cheng W; Liu CH

2009-04-01

66

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

Science.gov (United States)

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. PMID:24031739

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

2011-09-01

67

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

Scientific Electronic Library Online (English)

Full Text Available Abstract in english 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.

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

2011-09-01

68

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

UK PubMed Central (United Kingdom)

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.

Cavalli LS; Romano LA; Marins LF; Abreu PC

2011-07-01

69

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

Directory of Open Access Journals (Sweden)

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

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

2011-01-01

70

Pathogenesis of a Thai strain of white spot syndrome virus (WSSV) in juvenile, specific pathogen-free Litopenaeus vannamei  

Digital Repository Infrastructure Vision for European Research (DRIVER)

White spot syndrome virus (WSSV) causes disease and mortality in cultured and wild shrimp. A standardized WSSV oral inoculation procedure was used in specific pathogen-free (SPF) Litopenaeus vannamei (also called Penaeus vannamei) to determine the primary sites of replication (portal o...

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

71

Characterization of White Spot Syndrome Virus Envelope Protein VP51A and Its Interaction with Viral Tegument Protein VP26?  

Digital Repository Infrastructure Vision for European Research (DRIVER)

In this study, we characterize a novel white spot syndrome virus (WSSV) structural protein, VP51A (WSSV-TW open reading frame 294), identified from a previous mass spectrometry study. Temporal-transcription analysis showed that vp51A is expressed in the late stage of WSSV infection. Gene structure a...

Chang, Yun-Shiang; Liu, Wang-Jing; Chou, Tsung-Lu; Lee, Yuan-Ting; Lee, Tai-Lin; Huang, Wei-Tung; Kou, Guang-Hsiung

72

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

Digital Repository Infrastructure Vision for European Research (DRIVER)

Our aim was to isolate and characterize white spot syndrome virus (WSSV)-binding proteins from shrimp. After a blot of shrimp hemocyte membrane proteins was overlaid with a recombinant WSSV envelope protein (rVP28), the reactive bands on the blot were detected using anti-VP28 antibody. Among three m...

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

73

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

Digital Repository Infrastructure Vision for European Research (DRIVER)

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 feede...

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

74

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

Digital Repository Infrastructure Vision for European Research (DRIVER)

Although the Janus kinase-signal transducer and activator of transcription (JAK-STAT) signaling pathway is part of the antiviral response in arthropods such as Drosophila, here we show that white spot syndrome virus (WSSV) uses a shrimp STAT as a transcription factor to enhance viral gene expression...

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

75

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

Digital Repository Infrastructure Vision for European Research (DRIVER)

High temperature (32 to 33°C) has been shown to reduce mortality in white spot syndrome virus (WSSV)-infected shrimps, but the mechanism still remains unclear. Here we show that in WSSV-infected shrimps cultured at 32°C, transcriptional levels of representative immediate-early, early, and late genes...

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

76

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

Science.gov (United States)

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. PMID:23384051

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

2013-02-06

77

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

Directory of Open Access Journals (Sweden)

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.

Lu Liqun; Wang Hai; Manopo Ivanus; Yu Li; Kwang Jimmy

2005-01-01

78

[Diagnosis of white spot syndrome virus in farm crawfish in Anhui province and its epidemiological source].  

UK PubMed Central (United Kingdom)

OBJECTIVE: The aims of this study were to find out the pathogen causing high mortality of the crawfish population in Anhui over the past years. To provide the strategy for controlling the spread of the disease, an investigation was conducted to trace the origin of the pathogen. METHODS: Crawfish samples were tested by nested PCR, to diagnose the White Spot Syndrome Virus (WSSV). Healthy crawfish were inoculated with the supernatant of tissue homogenates. The viral suspension was purified using sucrose density gradients. The gill samples infected with purified virus were then subject to the transmission electron microscopy (TEM). The possible sources, including pond water, feedstuff, crabs, water plants, were investigated. RESULTS: Crawfish samples and commercial feeds exhibited all positive for WSSV. Challenge model on healthy crawfish recovered similar symptoms as the naturally infected ones on 7 to 9 days after inoculation. The viral particles were observed under TEM. CONCLUSION: Our findings indicated that WSSV was the causative agent that led to high mortality in the crawfish population in this area. These results demonstrated that the exotic viruses are derived from the regions where the frozen feeds were contaminated with WSSV-infected shell debris.

Pan Z; Yang Z; Lu C

2013-05-01

79

A novel JNK from Litopenaeus vannamei involved in white spot syndrome virus infection.  

UK PubMed Central (United Kingdom)

The c-Jun N-terminal kinase (JNK), a member of MAP kinases, is a serine/threonine-specific protein kinase which responds to extracellular stimuli and regulate various cellular activities. It is well documented in innate immune responses and reported to be involved in various viral infections of mammals. In present study, we cloned JNK homolog in a crustacean, Litopenaeus vannamei (designated as LvJNK) and studied its role in white spot syndrome virus (WSSV) infection. Sequence analysis displayed that LvJNK shared high similarity with other members of the JNK subfamily, including the conserved TPY motif and serine/threonine protein kinase (S_TKc) domain. Western blot analysis showed that the activation of LvJNK took place in WSSV infection. LvJnk silencing mediated by specific dsRNA in shrimps could significantly inhibit the proliferation of the virus. Moreover, inhibition of shrimp JNK signaling pathway by specific inhibitor resulted in the reduction of WSSV replication and the delay of WSSV gene transcription. These results indicate for the first time that shrimp JNK is activated in response to WSSV infection and WSSV could benefit from JNK activation. It may facilitate our understanding of the molecular mechanism of virus infection and provided a potential target for preventing the WSSV infection.

Shi H; Yan X; Ruan L; Xu X

2012-07-01

80

Role of chymotrypsin-like serine proteinase in white spot syndrome virus infection in Fenneropenaeus chinensis.  

UK PubMed Central (United Kingdom)

White spot syndrome virus (WSSV) caused a great economic loss in shrimp aquaculture. Although great efforts have been undertaken to characterize the virus disease during the last two decades, there are still lack of effective methods to prevent or cure it. In this study, we investigated the transcriptional expression profiles of 18 key immune-related genes in the Chinese shrimp Fenneropenaeus chinensis which was severely infected by WSSV. We found that the expression levels of 6 genes including chymotrypsin-like serine proteinase (CH-SPase), heat shock protein 70 cognate (HSP70), penaeidin (PEN), peroxinectin (PO), proliferating cell nuclear antigen (PCNA) and argonaute (AGO) changed significantly, while the expression of the other 12 genes had no significant changes compared to the control group. Among the 6 screened genes, CH-SPase showed significantly up-regulation, while the other 5 ones were significantly down-regulated. Knockdown of the expression of CH-SPase in WSSV-infected Chinese shrimp reduced the copy number of WSSV and delayed cumulative mortalities, suggesting that CH-SPase is important for WSSV infection. This study will be helpful to control the disease in shrimp caused by WSSV.

Xue S; Yang W; Sun J

2013-02-01

 
 
 
 
81

Role of chymotrypsin-like serine proteinase in white spot syndrome virus infection in Fenneropenaeus chinensis.  

Science.gov (United States)

White spot syndrome virus (WSSV) caused a great economic loss in shrimp aquaculture. Although great efforts have been undertaken to characterize the virus disease during the last two decades, there are still lack of effective methods to prevent or cure it. In this study, we investigated the transcriptional expression profiles of 18 key immune-related genes in the Chinese shrimp Fenneropenaeus chinensis which was severely infected by WSSV. We found that the expression levels of 6 genes including chymotrypsin-like serine proteinase (CH-SPase), heat shock protein 70 cognate (HSP70), penaeidin (PEN), peroxinectin (PO), proliferating cell nuclear antigen (PCNA) and argonaute (AGO) changed significantly, while the expression of the other 12 genes had no significant changes compared to the control group. Among the 6 screened genes, CH-SPase showed significantly up-regulation, while the other 5 ones were significantly down-regulated. Knockdown of the expression of CH-SPase in WSSV-infected Chinese shrimp reduced the copy number of WSSV and delayed cumulative mortalities, suggesting that CH-SPase is important for WSSV infection. This study will be helpful to control the disease in shrimp caused by WSSV. PMID:23142726

Xue, Shuxia; Yang, Weijun; Sun, Jinsheng

2012-11-07

82

Molecular detection of three shrimp viruses and genetic variation of white spot syndrome virus in Hainan Province, China, in 2007.  

Science.gov (United States)

Abstract White spot syndrome virus (WSSV), Taura syndrome virus (TSV) and infectious hypodermal and haematopoietic necrosis virus (IHHNV) have been responsible for major pandemics affecting the shrimp farming industry. Shrimp samples were collected from eight farms in Hainan Province, China, during 2007 and analysed by polymerase chain reaction (PCR) or reverse transcriptase PCR methods to determine the prevalence of these viruses. From the eight sampling locations, only samples from one farm did not show any indication of infection with WSSV, TSV or IHHNV, while samples from one additional farm exhibited evidence of infection with TSV only. Surprisingly, evidence of co-infection with TSV and IHHNV was found among samples at two farms while evidence of co-infection with all three viruses (WSSV, TSV and IHHNV) was detected among shrimp samples at four farms. To further elucidate the molecular characteristics of WSSV in China, we further analysed genomic features of WSSV isolates based on the ORF23/24 variable region. From these data, we identified two novel WSSV strains which contain nucleotide deletions of 5657 and 11093 bp, respectively, when compared with the largest WSSV-TW isolate. PMID:19490390

Tan, Y; Xing, Y; Zhang, H; Feng, Y; Zhou, Y; Shi, Z-L

2009-05-28

83

Molecular detection of three shrimp viruses and genetic variation of white spot syndrome virus in Hainan Province, China, in 2007.  

UK PubMed Central (United Kingdom)

Abstract White spot syndrome virus (WSSV), Taura syndrome virus (TSV) and infectious hypodermal and haematopoietic necrosis virus (IHHNV) have been responsible for major pandemics affecting the shrimp farming industry. Shrimp samples were collected from eight farms in Hainan Province, China, during 2007 and analysed by polymerase chain reaction (PCR) or reverse transcriptase PCR methods to determine the prevalence of these viruses. From the eight sampling locations, only samples from one farm did not show any indication of infection with WSSV, TSV or IHHNV, while samples from one additional farm exhibited evidence of infection with TSV only. Surprisingly, evidence of co-infection with TSV and IHHNV was found among samples at two farms while evidence of co-infection with all three viruses (WSSV, TSV and IHHNV) was detected among shrimp samples at four farms. To further elucidate the molecular characteristics of WSSV in China, we further analysed genomic features of WSSV isolates based on the ORF23/24 variable region. From these data, we identified two novel WSSV strains which contain nucleotide deletions of 5657 and 11093 bp, respectively, when compared with the largest WSSV-TW isolate.

Tan Y; Xing Y; Zhang H; Feng Y; Zhou Y; Shi ZL

2009-09-01

84

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

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

Ruangsri, J.; Phongdara, A.; Prasertsan, P.; Supamattaya, K.

2005-01-01

85

Viral interference between infectious hypodermal and hematopoietic necrosis virus and white spot syndrome virus in Litopenaeus vannamei.  

UK PubMed Central (United Kingdom)

White spot syndrome virus (WSSV) is highly virulent and has caused significant production losses to the shrimp culture industry over the last decade. Infectious hypodermal and hematopoietic necrosis virus (IHHNV) also infects penaeid shrimp and, while being less important than WSSV, remains a major cause of significant production losses in Litopenaeus vannamei (also called Penaeus vannamei) and L. stylirostris (also called Penaeus stylirostris). These 2 viruses and their interactions were previously investigated in L. stylirostris. We report here laboratory challenge studies carried out to determine if viral interference between IHHNV and WSSV also occurs in L. vannamei, and it was found that experimental infection with IHHNV induced a significant delay in mortality following WSSV challenge. L. vannamei infected per os with IHHNV were challenged with WSSV at 0, 10, 20, 30, 40 and 50 d post-infection. Groups of naïve shrimp infected with WSSV alone died in 3 d whereas shrimp pre-infected with IHHNV for 30, 40 or 50 d died in 5 d. Real-time PCR analysis showed that the delay correlated to the IHHNV load and that WSSV challenge induced a decrease in IHHNV load, indicating some form of competition between the 2 viruses.

Bonnichon V; Lightner DV; Bonami JR

2006-10-01

86

The detection of White Spot Syndrome Virus (WSSV) and Yellow Head Virus (YHV) in imported commodity shrimp.  

UK PubMed Central (United Kingdom)

Transmission of exotic pathogens occurs through a variety of means, including migration with humans and animals, rapid transit by land, sea or air or through the shipment of infected frozen food products. White Spot Syndrome Virus (WSSV) and Yellow Head Virus (YHV) have caused mass mortalities of cultured shrimp in Asia beginning in 1992. In 1995, these viruses appeared for the first time in the Western Hemisphere causing high mortalities in farm reared shrimp in Texas, USA. The purpose of this study was to determine if WSSV and YHV are present in frozen shrimp products imported into the United States from Asia. Infectivity assays, transmission electron microscopy (TEM), and polymerase chain reaction (PCR) showed these viruses were detectable and infectious in frozen shrimp imports. Frozen shrimp were used to infect indicator shrimp (Penaeus stylirostris) which resulted in mortalities. The cause of these mortalities was determined by histology and TEM to be by YHV. PCR confirmed the presence of WSSV in the frozen, purchased products. The results from this study indicate that exotic shrimp pathogens can be transmitted via imported frozen products.

Nunan LM; Poulos BT; Lightner DV

1998-01-01

87

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

Science.gov (United States)

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. PMID:18023170

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

2007-10-16

88

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

UK PubMed Central (United Kingdom)

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.

Clark KF; Greenwood SJ; Acorn AR; Byrne PJ

2013-09-01

89

Evidence for cell apoptosis suppressing white spot syndrome virus replication in Procambarus clarkii at high temperature.  

Science.gov (United States)

In shrimp, higher water temperatures (~32°C) can suppress the ability of white spot syndrome virus (WSSV) to replicate and cause mortality, but the mechanisms remain unclear. To investigate whether cell apoptosis might be involved, a Tdt-mediated dUTP nick-end label (TUNEL) method was used to assess levels of chromosomal DNA fragmentation in hepatopancreas and gill cells of Procambarus clarkii crayfish infected with WSSV and maintained at either 32 ± 1°C or 24 ± 1°C. Based on relative cell numbers with yellow-green colored TUNEL-positive nuclei, the apoptotic index was elevated in WSSV-infected crayfish maintained at 32°C. In gill tissue sections examined by transmission electron microscope, cells with nuclei displaying apoptotic bodies or marginated, condensed and fragmented chromatin without concurrent cell cytoplasm damage were also more prevalent. Flow cytometry sorting of annexin-stained cells showed apoptosis to be most prevalent in granular haemocytes, and assays for caspase-3 activity showed it to be most elevated in hepatopancreas tissue. Despite these indicators of cell apoptosis but consistent with WSSV replication being restricted at elevated temperatures, no increases in transcription of the viral anti-apoptosis genes ORF390 and ORF222 were detected by RT-PCR in shrimp maintained at 32°C, possibly due to the elevated levels of cellular apoptosis. PMID:23209074

Wu, Xiao-Guo; Xiong, Hai-Tao; Wang, Yi-Zhen; Du, Hua-Hua

2012-12-01

90

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

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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.

Wu Chenglin; Yang Feng

2006-01-01

91

BAX inhibitor-1 silencing suppresses white spot syndrome virus replication in red swamp crayfish, Procambarus clarkii.  

Science.gov (United States)

BAX inhibitor-1 (BI-1) was originally described as an anti-apoptotic protein in both animal and plant cells. BI-1 overexpression suppresses ER stress-induced apoptosis in animal cells. Inhibition of BI-1 activity could induce the cell death in mammals and plants. However, the function of BI-1 in crustacean immunity was unclear. In this paper, the full-length cDNA of a BI-1 protein in red swamp crayfish, Procambarus clarkii (PcBI-1) was cloned and its expression profiles in normal and infected crayfish were analyzed. The results showed that PcBI-1 was expressed in hemocytes, heart, hepatopancreas, gills, stomach, and intestines of the crayfish and was upregulated after challenged with Vibrio anguillarum and with white spot syndrome virus (WSSV). To determine the function of PcBI-1 in the innate immunity of the crayfish, the RNA interference against PcBI-1 was performed and the results indicated the hemocyte programmed cell death rate was increased significantly and WSSV replication was declined after PcBI-1 knocked down. Altogether, PcBI-1 plays an anti-apoptotic role, wherein high PcBI-1 expression suppresses programmed cell death, which is beneficial for WSSW replication in crayfish. PMID:23583724

Du, Zhi-Qiang; Lan, Jiang-Feng; Weng, Yu-Ding; Zhao, Xiao-Fan; Wang, Jin-Xing

2013-04-10

92

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

Science.gov (United States)

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

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

2008-08-01

93

Detection of white spot syndrome virus by polymerase chain reaction performed under insulated isothermal conditions.  

Science.gov (United States)

Aiming to develop a rapid, low-cost, and user-friendly system for the diagnosis of white spot syndrome virus (WSSV), a PCR assay performed in capillary tubes under insulated isothermal conditions (iiPCR assay) was established on the basis of Rayleigh-Benard convection. WSSV amplicons were generated reproducibly within 30 min from a target sequence-containing plasmid in an iiPCR device, in which a special polycarbonate capillary tube (R-tube™) was heated isothermally by a copper ring attached to its bottom and shielded by a thermal baffle around its upper half. Furthermore, WSSV-specific amplicons were produced from nucleic acid extracts of WSSV-infected Penaeus vannamei in the WSSV iiPCR assay, with sensitivity comparable to that of an OIE-certified commercial nested PCR kit (IQ2000™ WSSV Detection and Prevention System). Specificity of the WSSV iiPCR assay was demonstrated as no amplicons were generated from shrimp genomic DNA, and IHHNV, MBV, and HPV DNA. iiPCR has a potential as a low-cost method for sensitive, specific and rapid detection of pathogens. PMID:22326658

Tsai, Yun-Long; Lin, Yu-Chan; Chou, Pin-Hsing; Teng, Ping-Hua; Lee, Pei-Yu

2012-02-04

94

Detection of white spot syndrome virus by polymerase chain reaction performed under insulated isothermal conditions.  

UK PubMed Central (United Kingdom)

Aiming to develop a rapid, low-cost, and user-friendly system for the diagnosis of white spot syndrome virus (WSSV), a PCR assay performed in capillary tubes under insulated isothermal conditions (iiPCR assay) was established on the basis of Rayleigh-Benard convection. WSSV amplicons were generated reproducibly within 30 min from a target sequence-containing plasmid in an iiPCR device, in which a special polycarbonate capillary tube (R-tube™) was heated isothermally by a copper ring attached to its bottom and shielded by a thermal baffle around its upper half. Furthermore, WSSV-specific amplicons were produced from nucleic acid extracts of WSSV-infected Penaeus vannamei in the WSSV iiPCR assay, with sensitivity comparable to that of an OIE-certified commercial nested PCR kit (IQ2000™ WSSV Detection and Prevention System). Specificity of the WSSV iiPCR assay was demonstrated as no amplicons were generated from shrimp genomic DNA, and IHHNV, MBV, and HPV DNA. iiPCR has a potential as a low-cost method for sensitive, specific and rapid detection of pathogens.

Tsai YL; Lin YC; Chou PH; Teng PH; Lee PY

2012-04-01

95

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

UK PubMed Central (United Kingdom)

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

Feng S; Feng W; Zhao L; Gu H; Li Q; Shi K; Guo S; Zhang N

2013-10-01

96

BAX inhibitor-1 silencing suppresses white spot syndrome virus replication in red swamp crayfish, Procambarus clarkii.  

UK PubMed Central (United Kingdom)

BAX inhibitor-1 (BI-1) was originally described as an anti-apoptotic protein in both animal and plant cells. BI-1 overexpression suppresses ER stress-induced apoptosis in animal cells. Inhibition of BI-1 activity could induce the cell death in mammals and plants. However, the function of BI-1 in crustacean immunity was unclear. In this paper, the full-length cDNA of a BI-1 protein in red swamp crayfish, Procambarus clarkii (PcBI-1) was cloned and its expression profiles in normal and infected crayfish were analyzed. The results showed that PcBI-1 was expressed in hemocytes, heart, hepatopancreas, gills, stomach, and intestines of the crayfish and was upregulated after challenged with Vibrio anguillarum and with white spot syndrome virus (WSSV). To determine the function of PcBI-1 in the innate immunity of the crayfish, the RNA interference against PcBI-1 was performed and the results indicated the hemocyte programmed cell death rate was increased significantly and WSSV replication was declined after PcBI-1 knocked down. Altogether, PcBI-1 plays an anti-apoptotic role, wherein high PcBI-1 expression suppresses programmed cell death, which is beneficial for WSSW replication in crayfish.

Du ZQ; Lan JF; Weng YD; Zhao XF; Wang JX

2013-07-01

97

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

Science.gov (United States)

Abstract Aims: In this study we identified viral gene targets of the important redox regulator thioredoxin (Trx), and explored in depth how Trx interacts with the immediate early gene #1 (IE1) of the white spot syndrome virus (WSSV). Results: In a pull-down assay, we found that recombinant Trx bound to IE1 under oxidizing conditions, and a coimmunoprecipitation assay showed that Trx bound to WSSV IE1 when the transfected cells were subjected to oxidative stress. A pull-down assay with Trx mutants showed that no IE1 binding occurred when cysteine 62 was replaced by serine. Electrophoretic mobility shift assay (EMSA) showed that the DNA binding activity of WSSV IE1 was downregulated under oxidative conditions, and that Penaeus monodon Trx (PmTrx) restored the DNA binding activity of the inactivated, oxidized WSSV IE1. Another EMSA experiment showed that IE1's Cys-X-X-Cys motif and cysteine residue 55 were necessary for DNA binding. Measurement of the ratio of reduced glutathione to oxidized glutathione (GSH/GSSG) in WSSV-infected shrimp showed that oxidative stress was significantly increased at 48?h postinfection. The biological significance of Trx was also demonstrated in a double-strand RNA Trx knockdown experiment where suppression of shrimp Trx led to significant decreases in mortality and viral copy numbers. Innovation and Conclusion: WSSV's pathogenicity is enhanced by the virus' use of host Trx to rescue the DNA binding activity of WSSV IE1 under oxidizing conditions. Antioxid. Redox Signal. 17, 914–926.

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

2012-01-01

98

TAT-mediated oral subunit vaccine against white spot syndrome virus in crayfish.  

UK PubMed Central (United Kingdom)

White spot syndrome virus is a highly pathogenic virus that infects crayfish and other crustaceans. VP28 is one of its major envelope proteins, and plays a crucial role in viral infection. Cell-penetrating peptides are short peptides that facilitate cellular uptake of various molecular cargoes, and one well known example is TAT peptide from HIV-1 TAT protein. In this study, recombinant plasmids were constructed and transformed into Escherichia coli strain BL21 (DE3) to express TAT-VP28, VP28, TAT-VP28-EGFP and VP28-EGFP fusion proteins. Enzyme-linked immunosorbent assay (ELISA) and flow cytometry methods were used to confirm that TAT fusion proteins can translocate from the intestine to the hemolymph of the crayfish Cambarus clarkii. After immunization, activities of phenoloxidase and superoxide dismutase were analyzed, and it was found that rTAT-VP28 produced the most pronounced increase in both C. clarkii were vaccinated by oral administration of rTAT-VP28 and rVP28 for 7 and 14 days, and rTAT-VP28 resulted in the highest relative percent survival (RPS) (63.3% at 7 days, and 67.8% at 14 days), compared with rVP28 (44.4% at 7 days, and 53.6% at 14 days) following challenge with WSSV after the last day of feeding. This study reports the use of TAT-derived peptide as an oral delivery method of a subunit vaccine against WSSV in C. clarkii.

Zhang Y; Ning JF; Qu XQ; Meng XL; Xu JP

2012-04-01

99

White spot syndrome virus and infectious hypodermal and hematopoietic necrosis virus simultaneous diagnosis by miniarray system with colorimetry detection.  

Science.gov (United States)

Highly sensitive and specific diagnostic tools are essential for monitoring the health status of farmed species. After the development of genomic probe diagnostic systems in the 1990s, followed by PCR-based systems, a miniarray method has been developed allowing one-step multiple detection. The miniarray method was developed to enable the accessibility of powerful array technology. To use this system, hybridisation and washing process were modified, resulting into a significant increase in the test's rapidity and sensitivity. With miniarray technology, hybridisation time is reduced to 20 min, whereas other methods require a longer hybridisation time. Hybridisation of the PCR product on a nylon membrane and revelation of the hybrids by an antibody increase considerably the ability of pathogen's detection. A first application is developed for the diagnosis of two specific viruses which are, by their geographical range and their impact on the production, very important in shrimp pathology, namely, the White Spot Syndrome Virus (WSSV) and the Infectious Hypodermal and Hematopoietic Necrosis Virus (IHHNV). PMID:12270652

Quéré, Ronan; Commes, Thérèse; Marti, Jacques; Bonami, Jean Robert; Piquemal, David

2002-09-01

100

White spot syndrome virus and infectious hypodermal and hematopoietic necrosis virus simultaneous diagnosis by miniarray system with colorimetry detection.  

UK PubMed Central (United Kingdom)

Highly sensitive and specific diagnostic tools are essential for monitoring the health status of farmed species. After the development of genomic probe diagnostic systems in the 1990s, followed by PCR-based systems, a miniarray method has been developed allowing one-step multiple detection. The miniarray method was developed to enable the accessibility of powerful array technology. To use this system, hybridisation and washing process were modified, resulting into a significant increase in the test's rapidity and sensitivity. With miniarray technology, hybridisation time is reduced to 20 min, whereas other methods require a longer hybridisation time. Hybridisation of the PCR product on a nylon membrane and revelation of the hybrids by an antibody increase considerably the ability of pathogen's detection. A first application is developed for the diagnosis of two specific viruses which are, by their geographical range and their impact on the production, very important in shrimp pathology, namely, the White Spot Syndrome Virus (WSSV) and the Infectious Hypodermal and Hematopoietic Necrosis Virus (IHHNV).

Quéré R; Commes T; Marti J; Bonami JR; Piquemal D

2002-09-01

 
 
 
 
101

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

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Shrimp subcuticular epithelial cells are the initial and major targets of white spot syndrome virus (WSSV) infection. Proteomic studies of WSSV-infected subcuticular epithelium of Penaeus monodon were performed through two approaches, namely, subcellular fractionation coupled with shotgun proteomics...

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

102

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

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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.

Sutee WONGMANEEPRATEEP; Puttharat BAOPRASERTKUL; Piyanuch PROMPAMORN; Kanittada THONGKAO; Chalor LIMSUWAN; Niti CHUCHIRD

2010-01-01

103

Expression, Purification, Crystallization of Two Major Envelope Proteins from White Spot Syndrome Virus  

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White spot syndrome virus (WSSV) is a major virulent pathogen known to infect penaeid shrimp and other crustaceans. VP26 and VP28, two major envelope proteins from WSSV, have been identified and overexpressed in Escherichia coli. In order to facilitate purification and crystallization, predicted N-terminal transmembrane regions of approximately 35 amino acids have been truncated from both VP26 and VP28. Truncated VP26 and VP28 and their corresponding SeMet-labelled proteins were purified and the SeMet proteins were crystallized by the hanging-drop vapor-diffusion method. Crystals of SeMet-labelled VP26 were obtained using a reservoir consisting of 0.1 M citric acid pH 3.5, 3.0 M sodium chloride and 1%(w/v) polyethylene glycol 3350, whereas SeMet VP28 was crystallized using a reservoir solution consisting of 25% polyethylene glycol 8000, 0.2 M calcium acetate, 0.1 M Na HEPES pH 7.5 and 1.5%(w/v) 1,2,3-heptanetriol. Crystals of SeMet-labelled VP26 diffract to 2.2 {angstrom} resolution and belong to space group R32, with unit-cell parameters a = b = 73.92, c = 199.31 {angstrom}. SeMet-labelled VP28 crystallizes in space group P2{sub 1}2{sub 1}2{sub 1}, with unit-cell parameters a = 105.33, b = 106.71, c = 200.37 {angstrom}, and diffracts to 2.0 {angstrom} resolution.

Tang,X.; Hew, C.

2007-01-01

104

Shrimp MyD88 responsive to bacteria and white spot syndrome virus.  

UK PubMed Central (United Kingdom)

The myeloid differentiation factor 88 (MyD88) is an important adapter protein which links members of the toll-like receptor (TLR) to the downstream components to activate related signaling pathways. In the present study, a MyD88 homolog (FcMyD88) was cloned from penaeid shrimp Fenneropenaeus chinensis. The ORF of FcMyD88 consisted of 1434 bp encoding a polypeptide of 477 amino acids which contains a death domain (DD) and a typical TLR and interleukin-1 receptor (IL-1R)-related (TIR) domain. Homology analysis revealed that the predicted amino acid (aa) sequence of FcMyD88 shared high similarities with a variety of previously reported MyD88s. The time-dependent expression patterns of FcMyD88 in cephalothoraxes of shrimp injected with Vibrio anguillarum (Gram-negative bacteria, G(-)), Micrococcus lysodeikticu (Gram-positive bacteria, G(+)) and white syndrome spot virus (WSSV) were analyzed at transcription and protein level by real-time PCR and western blotting, respectively. The expression level of FcMyD88 mRNA was significantly up-regulated at one hour (h), 12 h and 24 h after stimulation with both V. anguillarum and M. lysodeikticu. The expression level of FcMyD88 protein was 2-fold up-regulated at 12 h post injection (hpi) of inactivated V. anguillarum while it didn't change after M. lysodeikticu injection during this period. After WSSV injection, the expression level of FcMyD88 mRNA remained relatively constant, while the FcMyD88 protein was significantly up-regulated at 12 and 24 hpi. These results suggested that the MyD88-dependent signaling pathway could be involved in the defense of both bacteria and WSSV infection.

Wen R; Li F; Sun Z; Li S; Xiang J

2013-02-01

105

Shrimp MyD88 responsive to bacteria and white spot syndrome virus.  

Science.gov (United States)

The myeloid differentiation factor 88 (MyD88) is an important adapter protein which links members of the toll-like receptor (TLR) to the downstream components to activate related signaling pathways. In the present study, a MyD88 homolog (FcMyD88) was cloned from penaeid shrimp Fenneropenaeus chinensis. The ORF of FcMyD88 consisted of 1434 bp encoding a polypeptide of 477 amino acids which contains a death domain (DD) and a typical TLR and interleukin-1 receptor (IL-1R)-related (TIR) domain. Homology analysis revealed that the predicted amino acid (aa) sequence of FcMyD88 shared high similarities with a variety of previously reported MyD88s. The time-dependent expression patterns of FcMyD88 in cephalothoraxes of shrimp injected with Vibrio anguillarum (Gram-negative bacteria, G(-)), Micrococcus lysodeikticu (Gram-positive bacteria, G(+)) and white syndrome spot virus (WSSV) were analyzed at transcription and protein level by real-time PCR and western blotting, respectively. The expression level of FcMyD88 mRNA was significantly up-regulated at one hour (h), 12 h and 24 h after stimulation with both V. anguillarum and M. lysodeikticu. The expression level of FcMyD88 protein was 2-fold up-regulated at 12 h post injection (hpi) of inactivated V. anguillarum while it didn't change after M. lysodeikticu injection during this period. After WSSV injection, the expression level of FcMyD88 mRNA remained relatively constant, while the FcMyD88 protein was significantly up-regulated at 12 and 24 hpi. These results suggested that the MyD88-dependent signaling pathway could be involved in the defense of both bacteria and WSSV infection. PMID:23238389

Wen, Rong; Li, Fuhua; Sun, Zheng; Li, Shihao; Xiang, Jianhai

2012-12-10

106

A single-step multiplex PCR for simultaneous detection of white spot syndrome virus and infectious hypodermal and haematopoietic necrosis virus in penaeid shrimp.  

UK PubMed Central (United Kingdom)

White spot syndrome virus (WSSV) and infectious hypodermal and haematopoietic necrosis virus (IHHNV) are the major viral pathogens of penaeid shrimp worldwide (Lightner & Redman 1998). Litopenaeus vannamei was introduced into China from the Americas, and quickly became widely cultured. Following its introduction, both IHHNV and WSSV have become important pathogens of cultured penaeid shrimp and have had a huge impact on the culture industry in China in recent years.

Yang B; Song XL; Huang J; Shi CY; Liu QH; Liu L

2006-05-01

107

A single-step multiplex PCR for simultaneous detection of white spot syndrome virus and infectious hypodermal and haematopoietic necrosis virus in penaeid shrimp.  

Science.gov (United States)

White spot syndrome virus (WSSV) and infectious hypodermal and haematopoietic necrosis virus (IHHNV) are the major viral pathogens of penaeid shrimp worldwide (Lightner & Redman 1998). Litopenaeus vannamei was introduced into China from the Americas, and quickly became widely cultured. Following its introduction, both IHHNV and WSSV have become important pathogens of cultured penaeid shrimp and have had a huge impact on the culture industry in China in recent years. PMID:16677320

Yang, B; Song, X-L; Huang, J; Shi, C-Y; Liu, Q-H; Liu, L

2006-05-01

108

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

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

RIA IKA MAHARANI; SURANTO; ZAFRAN

2005-01-01

109

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

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

He Fang; Kwang Jimmy

2008-01-01

110

Studies on the transmission of WSSV (white spot syndrome virus) in juvenile Marsupenaeus japonicus via marine microalgae.  

Science.gov (United States)

We studied the possible role that marine microalgae may play during the outbreaks of WSS (white spot syndrome). In order to elucidate the possibility of marine microalgae carrying WSSV (white spot syndrome virus), six marine microalgae (Isochrysis galbana, Skeletonema costatum, Chlorella sp., Heterosigma akashiwo, Scrippsiella trochoidea, Dunaliella salina) were co-cultured with adult Marsupenaeus japonicus infected with WSSV and were assayed daily by nested-PCR to study whether they could carry WSSV. Further experiments were conducted to investigate whether the virus carried by microalgae could re-infect juvenile M. japonicus. Results showed that all of the experimental microalgae, except H. akashiwo could carry WSSV, and among them, Chlorella sp. and S. trochoidea had the strongest WSSV-carrying ability. Unlike other invertebrate carriers of WSSV, the WSSV detections in microalgae, which were positive after 1 and 3 days, were negative after 10days of incubation. WSSV detection results in juvenile M. japonicus showed that the juvenile shrimp were re-infected by co-cultured Chlorella sp., although the juvenile M. japonicus carried so small an amount of WSSV that it could only be detected by nested-PCR. The results of this experiment suggest that microalgae might be one possible horizontal transmission pathway for WSSV. Further research, however, is required to better understand the factors behind the different carrying abilities and virus-carrying mechanisms of different microalgae. PMID:17383676

Liu, Bo; Yu, Zhiming; Song, Xiuxian; Guan, Yueqiang

2007-02-03

111

Studies on the transmission of WSSV (white spot syndrome virus) in juvenile Marsupenaeus japonicus via marine microalgae.  

UK PubMed Central (United Kingdom)

We studied the possible role that marine microalgae may play during the outbreaks of WSS (white spot syndrome). In order to elucidate the possibility of marine microalgae carrying WSSV (white spot syndrome virus), six marine microalgae (Isochrysis galbana, Skeletonema costatum, Chlorella sp., Heterosigma akashiwo, Scrippsiella trochoidea, Dunaliella salina) were co-cultured with adult Marsupenaeus japonicus infected with WSSV and were assayed daily by nested-PCR to study whether they could carry WSSV. Further experiments were conducted to investigate whether the virus carried by microalgae could re-infect juvenile M. japonicus. Results showed that all of the experimental microalgae, except H. akashiwo could carry WSSV, and among them, Chlorella sp. and S. trochoidea had the strongest WSSV-carrying ability. Unlike other invertebrate carriers of WSSV, the WSSV detections in microalgae, which were positive after 1 and 3 days, were negative after 10days of incubation. WSSV detection results in juvenile M. japonicus showed that the juvenile shrimp were re-infected by co-cultured Chlorella sp., although the juvenile M. japonicus carried so small an amount of WSSV that it could only be detected by nested-PCR. The results of this experiment suggest that microalgae might be one possible horizontal transmission pathway for WSSV. Further research, however, is required to better understand the factors behind the different carrying abilities and virus-carrying mechanisms of different microalgae.

Liu B; Yu Z; Song X; Guan Y

2007-06-01

112

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

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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 virus)infection 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.

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

2011-01-01

113

Anti-white spot syndrome virus activity of Ceriops tagal aqueous extract in giant tiger shrimp Penaeus monodon.  

UK PubMed Central (United Kingdom)

White spot syndrome virus (WSSV), the most contagious pathogen of cultured shrimp, causes mass mortality, leading to huge economic loss to the shrimp industry. The lack of effective therapeutic or prophylactic measures has aggravated the situation, necessitating the development of antiviral agents. With this objective, the antiviral activity in the aqueous extract of a mangrove plant Ceriops tagal in Penaeus monodon was evaluated. The Ceriops tagal aqueous extract (CTAE) was non-toxic to shrimps at 50 mg/ml when injected intramuscularly at a dosage of 10 ?L/animal (0.5 mg/animal) and showed a protective effect against WSSV at 30 mg/ml when mixed with WSSV suspension at a 1:1 ratio. When the extract was administered along with the diet and the animals were challenged orally, there was a dose-dependent increase in survival, culminating in 100 % survival at a concentration of 500 mg/kg body weight/day. Neither hypertrophied nuclei nor the viral envelope protein VP28 could be demonstrated in surviving shrimps using histology and indirect immunofluorescence histochemistry (IIFH), respectively. To elucidate the mode of action, the temporal expression of WSSV genes and shrimp immune genes, including antimicrobial peptides, was attempted. None of the viral genes were found to be expressed in shrimps that were fed with the extract and challenged or in those that were administered CTAE-exposed WSSV. The overall results suggest that the aqueous extract from C. tagal can protect P. monodon from white spot syndrome virus infection.

Sudheer NS; Philip R; Bright Singh IS

2012-09-01

114

Enhanced white spot syndrome virus (WSSV) detection sensitivity using monoclonal antibody specific to heterologously expressed VP19 envelope protein  

UK PubMed Central (United Kingdom)

The gene encoding the VP19 envelope protein of white spot syndrome virus (WSSV) was cloned into pMAL-C2 expression vector and transformed into the BL21 Escherichia coli strain. After induction, recombinant maltose binding protein (MBP)-VP19 fusion protein was produced, purified and electroeluted before use for immunization in Swiss mice for monoclonal antibody (MAb) production. MAbs specific to VP19 can be used to detect natural WSSV infection in Penaeus vannamei by dot blotting, western blotting and immunohistochemistry without cross-reaction to other shrimp tissues or other common shrimp viruses, including Taura syndrome virus (TSV), yellow head virus (YHV), Penaeus monodon nucleopolyhedrovirus (PemoNPV) formerly called monodon baculovirus (MBV) and P. monodon densovirus (PmDNV) previously called hepatopancreatic parvovirus (HPV). The detection sensitivity of the VP19-specific W25-8D MAb generated in this study was approximately 1.2fmol/spot of purified recombinant MBP-VP19 protein as determined by dot blotting while that of a VP28-specific W29 MAb obtained from a previous study was approximately 5fmol/spot. Combining MAbs specific for VP19 and VP28 resulted in two-fold higher sensitivity than use of either MAb alone. However, the sensitivity of the combined MAbs was 25,000 times lower than that of one-step PCR. Immunohistochemical analysis using MAbs specific to VP19 in WSSV-infected gill tissues and appendages demonstrated intense staining patterns in both the nucleus and cytoplasm compared to MAb specific to VP28. In conclusion, combination of VP19- and VP28-specific MAbs could confirm and enhance the sensitivity of WSSV detection in shrimp in various types of antibody-based assays.

Chaivisuthangkura Parin; Longyant Siwaporn; Rukpratanporn Sombat; Srisuk Chutima; Sridulyakul Pattarin; Sithigorngul Paisarn

2010-02-01

115

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

UK PubMed Central (United Kingdom)

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.

Thagun C; Srisala J; Sritunyalucksana K; Narangajavana J; Sojikul P

2012-09-01

116

White Spot (Early Tooth Decay)  

Science.gov (United States)

... TMJ Disorders Oral Cancer Dry Mouth Burning Mouth Tooth Decay See All Oral Health Oral Complications of ... Sorted by ... > Children's Oral Health White Spot Early tooth decay Share This Page Facebook External link – please ...

117

Improvement of immunodetection of white spot syndrome virus using a monoclonal antibody specific for heterologously expressed icp11.  

UK PubMed Central (United Kingdom)

The icp11 gene encoding the highly abundant DNA mimic protein of white spot syndrome virus (WSSV) was cloned into the pTYB1 and pGEX-6P-1 expression vectors and introduced into E. coli by transformation. After induction, C-terminally intein-tagged ICP11 (ICP11-intein) and N-terminally glutathione-S-transferase (GST)-tagged ICP11 (GST-ICP11) proteins with molecular masses of 64 and 35 kDa were obtained. These proteins were purified by SDS-PAGE and used for immunization of Swiss mice for monoclonal antibody (MAb) production. Two MAbs specific for ICP11 were selected; these MAbs can be used to detect natural WSSV infection in Penaeus vannamei by dot blotting, western blotting or immunohistochemistry without cross-reaction with other shrimp tissues or other common shrimp viruses. The detection sensitivity of the MAbs was approximately 0.7 fmole/spot of GST-ICP11 as determined by dot blotting. These MAbs showed stronger immunoreactivity than other MAbs from previous studies that are specific for VP28 and VP19. A combination of MAbs specific for ICP11, VP28 and VP19 increased the detection sensitivity of WSSV during early infection to a sensitivity 250 times lower than that of one-step PCR. Therefore, the MAbs specific for ICP11 could be used to confirm and enhance the detection sensitivity for WSSV infection in shrimp using various types of antibody-based assays.

Siriwattanarat R; Longyant S; Chaivisuthangkura P; Wangman P; Vaniksampanna A; Sithigorngul P

2013-05-01

118

Improvement of immunodetection of white spot syndrome virus using a monoclonal antibody specific for heterologously expressed icp11.  

Science.gov (United States)

The icp11 gene encoding the highly abundant DNA mimic protein of white spot syndrome virus (WSSV) was cloned into the pTYB1 and pGEX-6P-1 expression vectors and introduced into E. coli by transformation. After induction, C-terminally intein-tagged ICP11 (ICP11-intein) and N-terminally glutathione-S-transferase (GST)-tagged ICP11 (GST-ICP11) proteins with molecular masses of 64 and 35 kDa were obtained. These proteins were purified by SDS-PAGE and used for immunization of Swiss mice for monoclonal antibody (MAb) production. Two MAbs specific for ICP11 were selected; these MAbs can be used to detect natural WSSV infection in Penaeus vannamei by dot blotting, western blotting or immunohistochemistry without cross-reaction with other shrimp tissues or other common shrimp viruses. The detection sensitivity of the MAbs was approximately 0.7 fmole/spot of GST-ICP11 as determined by dot blotting. These MAbs showed stronger immunoreactivity than other MAbs from previous studies that are specific for VP28 and VP19. A combination of MAbs specific for ICP11, VP28 and VP19 increased the detection sensitivity of WSSV during early infection to a sensitivity 250 times lower than that of one-step PCR. Therefore, the MAbs specific for ICP11 could be used to confirm and enhance the detection sensitivity for WSSV infection in shrimp using various types of antibody-based assays. PMID:23242776

Siriwattanarat, Ruthairat; Longyant, Siwaporn; Chaivisuthangkura, Parin; Wangman, Pradit; Vaniksampanna, Akapon; Sithigorngul, Paisarn

2012-12-16

119

Effects of high salinity, high temperature and pH on capsid structure of white spot syndrome virus.  

Science.gov (United States)

The structural stability of white spot syndrome virus (WSSV) capsids at high salinity, high temperature and various pH values was studied. To obtain the viral capsids, the nucleocapsids were treated with high salinity. The results showed that high salinity treatment can cause the dissociation of VP15 and most of VP95 from the nucleocapsid, but there were no noticeable alterations in morphology and ultrastructure of the nucleocapsid and capsid. The capsids retained morphological integrity at temperatures 60°C. In addition, the capsids were relatively resistant to strong acid conditions and were tolerant to a broad pH range of 1 to 10. However, morphological change occurred at pH 10.5. The capsids broke up into small pieces at pH 11 and completely degraded in 0.1 and 1.0 M NaOH. These results indicated that the WSSV capsid is acid-stable and alkali-labile. PMID:23135144

Chen, Weiyu; Zhang, Heng; Gu, Li; Li, Fang; Yang, Feng

2012-11-01

120

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

International Nuclear Information System (INIS)

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

2005-09-15

 
 
 
 
121

Comparison of Protein Expression Profiles of the Hepatopancreas in Fenneropenaeus chinensis Challenged with Heat-inactivated Vibrio anguillarum and White Spot Syndrome Virus.  

UK PubMed Central (United Kingdom)

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.

Jiang H; Li F; Zhang J; Zhang J; Huang B; Yu Y; Xiang J

2013-09-01

122

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

International Nuclear Information System (INIS)

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

2009-09-01

123

Anti-white spot syndrome virus activity of Ceriops tagal aqueous extract in giant tiger shrimp Penaeus monodon.  

Science.gov (United States)

White spot syndrome virus (WSSV), the most contagious pathogen of cultured shrimp, causes mass mortality, leading to huge economic loss to the shrimp industry. The lack of effective therapeutic or prophylactic measures has aggravated the situation, necessitating the development of antiviral agents. With this objective, the antiviral activity in the aqueous extract of a mangrove plant Ceriops tagal in Penaeus monodon was evaluated. The Ceriops tagal aqueous extract (CTAE) was non-toxic to shrimps at 50 mg/ml when injected intramuscularly at a dosage of 10 ?L/animal (0.5 mg/animal) and showed a protective effect against WSSV at 30 mg/ml when mixed with WSSV suspension at a 1:1 ratio. When the extract was administered along with the diet and the animals were challenged orally, there was a dose-dependent increase in survival, culminating in 100 % survival at a concentration of 500 mg/kg body weight/day. Neither hypertrophied nuclei nor the viral envelope protein VP28 could be demonstrated in surviving shrimps using histology and indirect immunofluorescence histochemistry (IIFH), respectively. To elucidate the mode of action, the temporal expression of WSSV genes and shrimp immune genes, including antimicrobial peptides, was attempted. None of the viral genes were found to be expressed in shrimps that were fed with the extract and challenged or in those that were administered CTAE-exposed WSSV. The overall results suggest that the aqueous extract from C. tagal can protect P. monodon from white spot syndrome virus infection. PMID:22643833

Sudheer, N S; Philip, Rosamma; Bright Singh, I S

2012-05-29

124

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

Directory of Open Access Journals (Sweden)

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

Héctor Manuel Esparza Leal; Francisco Javier Magallón Barajas; Ricardo Pérez Enriquez; Ramón Casillas Hernández; Julio A. Cabanillas Ramos; Wenceslao Valenzuela Quiñónez

2012-01-01

125

The effect of raising water temperature to 33 °C in Penaeus vannamei juveniles at different stages of infection with white spot syndrome virus (WSSV)  

Digital Repository Infrastructure Vision for European Research (DRIVER)

This study investigated the effect of high water temperature (33 °C) at different stages of infection with a highly virulent and low virulent white spot syndrome virus strain (WSSV Thai-1 and WSSV Viet) in Penaeus vannamei juveniles. Shrimp were inoculated intramuscularly with eithe...

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

126

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

Digital Repository Infrastructure Vision for European Research (DRIVER)

To investigate whether specific-pathogen-free (SPF) shrimps are asymptomatic carriers of white spot syndrome virus (WSSV), we used a WSSV-specific DNA microarray to measure WSSV gene expression in SPF and WSSV-infected shrimps. Three WSSV genes were found to be relatively highly expressed in SPF shr...

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

127

Co-Interactive DNA-Binding between a Novel, Immunophilin-Like Shrimp Protein and VP15 Nucleocapsid Protein of White Spot Syndrome Virus  

Digital Repository Infrastructure Vision for European Research (DRIVER)

White spot syndrome virus (WSSV) is one of the most serious pathogens of penaeid shrimp. Although its genome has been completely characterized, the functions of most of its putative proteins are not yet known. It has been suggested that the major nucleocapsid protein VP15 is involved in packaging of...

Sangsuriya, Pakkakul; Senapin, Saengchan; Huang, Wei-Pang; Lo, Chu-Fang; Flegel, Timothy W.

128

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

Digital Repository Infrastructure Vision for European Research (DRIVER)

We show here that the white spot syndrome virus (WSSV) immediate-early protein IE1 interacts with the Penaeus monodon TATA box-binding protein (PmTBP) and that this protein-protein interaction occurs in the absence of any other viral or cellular proteins or nucleic acids, both in vitro and in vivo. ...

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

129

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

Digital Repository Infrastructure Vision for European Research (DRIVER)

White spot syndrome virus (WSSV) is a virulent pathogen known to infect various crustaceans. It has bacilliform morphology with a tail-like appendage at one end. The envelope consists of four major proteins. Envelope structural proteins play a crucial role in viral infection and are believed to be t...

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

130

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

Digital Repository Infrastructure Vision for European Research (DRIVER)

White spot syndrome virus (WSSV) is a major pathogen in shrimp aquaculture. VP9, a full-length protein of WSSV, encoded by open reading frame wsv230, was identified for the first time in the infected Penaeus monodon shrimp tissues, gill, and stomach as a novel, nonstructural protein by Western blott...

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

131

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

Digital Repository Infrastructure Vision for European Research (DRIVER)

The envelopment of the nucleocapsid is an important step in white spot syndrome virus (WSSV) assembly. Previous studies showed that VP26, a major envelope protein of WSSV, can interact with viral nucleocapsid. In this study, using the biotin label transfer technique, we found that the biotin label w...

Wan, Qiang; Xu, Limei; Yang, Feng

132

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

Digital Repository Infrastructure Vision for European Research (DRIVER)

We have characterized a white spot syndrome virus (WSSV) RING-H2-type protein, WSSV222, which is involved in ubiquitination. WSSV222 exhibits RING-H2-dependent E3 ligase activity in vitro in the presence of the specific conjugating enzyme UbcH6. Mutations in the RING-H2 domain abolished WSSV222-depe...

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

133

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

International Nuclear Information System (INIS)

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

2005-04-10

134

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

UK PubMed Central (United Kingdom)

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.

Huang HT; Leu JH; Huang PY; Chen LL

2012-01-01

135

Real-time quantitative loop-mediated isothermal amplification as a simple method for detecting white spot syndrome virus.  

UK PubMed Central (United Kingdom)

AIMS: White spot syndrome virus (WSSV) continues to be the most pathogenic virus among the crustacean aquaculture causing mass mortality. In the present study, we established a one-step, single tube, real-time accelerated loop-mediated isothermal amplification (real-time LAMP) for quantitative detection of WSSV. MATERIALS AND METHODS: A set of six specially designed primers that recognize eight distinct sequences of the target. The whole process can be completed in 1 h under isothermal conditions at 63 degrees C. Detection and quantification can be achieved by real-time monitoring in an inexpensive turbidimeter based on threshold time required for turbidity in the LAMP reaction. A standard curve was constructed by plotting viral titre against the threshold time (T(t)) using plasmid standards with high correlation coefficient (R(2) = 0.988). CONCLUSIONS: Sensitivity analysis using 10-fold dilutions (equivalent to 35 ng microl(-1) to 35 ag microl(-1)) of plasmid standards revealed this method is capable of detecting upto 100 copies of template DNA. Cross-reactivity analysis with DNA/cDNA of IHHNV, TSV, YHV-infected and healthy shrimp showed this method is highly specific for quantitative detection of WSSV. SIGNIFICANCE AND IMPACT OF THE STUDY: WSSV real-time LAMP assay appears to be precise, accurate and a valuable tool for the detection and quantification of WSSV in large field samples and epidemiological studies.

Mekata T; Sudhakaran R; Kono T; Supamattaya K; Linh NT; Sakai M; Itami T

2009-01-01

136

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.  

UK PubMed Central (United Kingdom)

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 (C(T)) values over a range of 1 to 10(5) copies of the viral genome. To control the variation in sampling and processing among samples, the shrimp beta-actin gene was amplified in parallel with the viral DNA. The C(T) values of IHHNV- and WSV-infected samples were used to determine absolute viral copy numbers from the standard C(T) curves of these viruses. For each virus and its beta-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.

Dhar AK; Roux MM; Klimpel KR

2001-08-01

137

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.  

Science.gov (United States)

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 (C(T)) values over a range of 1 to 10(5) copies of the viral genome. To control the variation in sampling and processing among samples, the shrimp beta-actin gene was amplified in parallel with the viral DNA. The C(T) values of IHHNV- and WSV-infected samples were used to determine absolute viral copy numbers from the standard C(T) curves of these viruses. For each virus and its beta-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

Dhar, A K; Roux, M M; Klimpel, K R

2001-08-01

138

F0ATP synthase b-chain of Litopenaeus vannamei involved in white spot syndrome virus infection.  

Science.gov (United States)

White Spot Syndrome Virus (WSSV) is one of the most common and distrous diseases for shrimp. In this study, we show that the protein VP292 that is a envelop protein of WSVV interacts with F0ATP synthase b-chain from Litopenaeus vannamei using far-western blot, ELISA, and indirect immunofluorescence analysis. Tissue distribution analysis of F0ATP synthase b-chain showed that it's transcription can be detected in muscle, hepatopancreas, intestine, hemocytes, lymphoid, and gills. Cellular localization of F0ATP synthase b-chain in shrimp hemocytes showed that F0ATP synthase b-chain was primarily located in the cytoplasm of hemocytes. The transcription levels of F0ATP synthase b-chain were significantly upregulated in intestine, hepatopancreas, hemocytes, and gills of WSSV-infected shrimp at 12 h after infection. Far-western, ELISA, and indirect immunefluorescence assay indicated that F0ATP synthase b-chain interacted with VP292. In the in vivo neutralization experiment, F0ATP synthase b-chain attained 18% protection rate of the shrimp challenged by WSSV. To the best of our knowledge, this is the first report to show that F0ATP synthase b-chain is involved in WSSV infection. PMID:23558437

Li, Qian; Liu, Qing-hui; Huang, Jie

2013-04-05

139

F0ATP synthase b-chain of Litopenaeus vannamei involved in white spot syndrome virus infection.  

UK PubMed Central (United Kingdom)

White Spot Syndrome Virus (WSSV) is one of the most common and distrous diseases for shrimp. In this study, we show that the protein VP292 that is a envelop protein of WSVV interacts with F0ATP synthase b-chain from Litopenaeus vannamei using far-western blot, ELISA, and indirect immunofluorescence analysis. Tissue distribution analysis of F0ATP synthase b-chain showed that it's transcription can be detected in muscle, hepatopancreas, intestine, hemocytes, lymphoid, and gills. Cellular localization of F0ATP synthase b-chain in shrimp hemocytes showed that F0ATP synthase b-chain was primarily located in the cytoplasm of hemocytes. The transcription levels of F0ATP synthase b-chain were significantly upregulated in intestine, hepatopancreas, hemocytes, and gills of WSSV-infected shrimp at 12 h after infection. Far-western, ELISA, and indirect immunefluorescence assay indicated that F0ATP synthase b-chain interacted with VP292. In the in vivo neutralization experiment, F0ATP synthase b-chain attained 18% protection rate of the shrimp challenged by WSSV. To the best of our knowledge, this is the first report to show that F0ATP synthase b-chain is involved in WSSV infection.

Li Q; Liu QH; Huang J

2013-08-01

140

Antigenic and immunogenic properties of truncated VP28 protein of white spot syndrome virus in Procambarus clarkii.  

Science.gov (United States)

Previous studies identify VP28 envelope protein of white spot syndrome virus (WSSV) as its main antigenic protein. Although implicated in viral infectivity, its functional role remains unclear. In the current study, we described the production of polyclonal antibodies to recombinant truncated VP28 proteins including deleted N-terminal (rVP28?N), C-terminal (rVP28?C) and middle (rVP28?M). In antigenicity assays, antibodies developed from VP28 truncations lacking the N-terminal or middle regions showed significantly lowered neutralization of WSSV in crayfish, Procambarus clarkii. Further immunogenicity analysis showed reduced relative percent survival (RPS) in crayfish vaccinating with these truncations before challenge with WSSV. These results indicated that N-terminal (residues 1-27) and middle region (residues 35-95) were essential to maintain the neutralizing linear epitopes of VP28 and responsible in eliciting immune response. Thus, it is most likely that these regions are exposed on VP28, and will be useful for rational design of effective vaccines targeting VP28 of WSSV. PMID:23178263

Du, Hua-Hua; Hou, Chong-Lin; Wu, Xiao-Guo; Xie, Rong-hui; Wang, Yi-Zhen

2012-11-23

 
 
 
 
141

High efficacy of white spot syndrome virus replication in tissues of freshwater rice-field crab, Paratelphusa hydrodomous (Herbst).  

Science.gov (United States)

An attempt was made to determine the replication efficiency of white spot syndrome virus (WSSV) of shrimp in different organs of freshwater rice-field crab, Paratelphusa hydrodomous (Herbst), using bioassay, PCR, RT-PCR, ELISA, Western blot and real-time PCR analyses, and also to use this crab instead of penaeid shrimp for the large-scale production of WSSV. This crab was found to be highly susceptible to WSSV by intramuscular injection. PCR and Western blot analyses confirmed the systemic WSSV infection in freshwater crab. The RT-PCR analysis revealed the expression of VP28 gene in different organs of infected crab. The indirect ELISA was used to quantify the VP28 protein in different organs of crab. It was found that there was a high concentration of VP28 protein in gill tissue, muscle, haemolymph and heart tissue. The copy number of WSSV in different organs of infected crab was quantified by real-time PCR, and the results revealed a steady increase in copy number in different organs of infected crab during the course of infection. The viral inoculum prepared from different organs of infected crab caused significant mortality in tiger prawn, Penaeus monodon (Fabricius). The results revealed that this crab can be used as an alternate host for WSSV replication and production. PMID:22943699

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

2012-09-03

142

Role of Penaeus monodon Kruppel-like factor (PmKLF) in infection by white spot syndrome virus.  

UK PubMed Central (United Kingdom)

Sp1-like proteins and Kruppel-like factors (KLFs) are highly related zinc-finger proteins that have crucial roles in transcription. One expressed sequence tag (EST, HPA-N-S01-EST0038) from shrimps is homologous to Sp1. This study reports the cloning and characteristics of a KLF from shrimp, Penaeus monodon (PmKLF). The full-length PmKLF cDNA is 1702 bp, encoding a polypeptide of 360 amino acids. Sequence analysis revealed that the sequence of PmKLF is similar to that of KLF11 in humans, mice and zebrafish. RT-PCR analysis indicated that PmKLF mRNA is expressed in all examined tissues. Additionally, immunofluorescence analysis revealed that GFP-KLF fusion protein is located in the nucleus as dots in an insect cell line, Sf9. Localization of PmKLF in the nucleus is also observed in the hemolymph from white spot syndrome virus (WSSV)-infected and WSSV-uninfected Litopenaeus vannamei. Knockdown of the expression of PmKLF transcript in WSSV-infected shrimp resulted in delayed cumulative mortalities, suggesting that PmKLF is important to WSSV infection. Moreover, inhibition of PmKLF expression reduced the copy number of WSSV and ie1 expression, revealing that PmKLF affects WSSV infection via interfering with ie1 expression.

Chang LK; Huang PH; Shen WT; Yang SH; Liu WJ; Lo CF

2012-01-01

143

Antigenic and immunogenic properties of truncated VP28 protein of white spot syndrome virus in Procambarus clarkii.  

UK PubMed Central (United Kingdom)

Previous studies identify VP28 envelope protein of white spot syndrome virus (WSSV) as its main antigenic protein. Although implicated in viral infectivity, its functional role remains unclear. In the current study, we described the production of polyclonal antibodies to recombinant truncated VP28 proteins including deleted N-terminal (rVP28?N), C-terminal (rVP28?C) and middle (rVP28?M). In antigenicity assays, antibodies developed from VP28 truncations lacking the N-terminal or middle regions showed significantly lowered neutralization of WSSV in crayfish, Procambarus clarkii. Further immunogenicity analysis showed reduced relative percent survival (RPS) in crayfish vaccinating with these truncations before challenge with WSSV. These results indicated that N-terminal (residues 1-27) and middle region (residues 35-95) were essential to maintain the neutralizing linear epitopes of VP28 and responsible in eliciting immune response. Thus, it is most likely that these regions are exposed on VP28, and will be useful for rational design of effective vaccines targeting VP28 of WSSV.

Du HH; Hou CL; Wu XG; Xie RH; Wang YZ

2013-01-01

144

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

UK PubMed Central (United Kingdom)

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.

Nathiga Nambi KS; Abdul Majeed S; Sundar Raj N; Taju G; Madan N; Vimal S; Sahul Hameed AS

2012-08-01

145

An investigation into occasional White Spot Syndrome Virus outbreak in traditional paddy cum prawn fields in India.  

UK PubMed Central (United Kingdom)

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.

Selvam DG; Mujeeb Rahiman KM; Mohamed Hatha AA

2012-01-01

146

Influence of acute salinity changes on biochemical, hematological and immune characteristics of Fenneropenaeus indicus during white spot syndrome virus challenge.  

UK PubMed Central (United Kingdom)

The present study reports the influence of salinity (5, 15, 25 and 35 g/L) on the biochemical and immune characteristics of Fenneropenaeus indicus challenged with 5. 5 × 10(4) copy number of white spot syndrome virus (WSSV). F. indicus that had been reared in 25 g/L, injected with WSSV and transferred to 5, 15, 25 (control) and 35 g/L were examined after 0-120 hrs for total hemocyte count (THC), phenoloxidase (PO) and respiratory burst (RB) activity and alkaline and acid phosphatase activities. It was concluded that F. indicus that had been transferred from 25 g/L to lower and higher salinity levels (5, 15 and 35 g/L) had poorer immune indices and decreased resistance against WSSV infection. After 120 hrs, the mortality rate in WSSV-injected F. indicus experimental groups (5 and 35 g/L) was significantly higher than for F. indicus exposed to 25 and 15 g/L salinities. During the experimental period (0-120 hrs), biochemical variables, namely total protein, carbohydrate, and lipid concentrations, were measured in hemolymph of both experimental and control groups. Acute salinity changes induced an increase in protein variations across the tested salinity ranges in shrimp. After 24 hrs, THC and PO activity decreased significantly whereas RB, alkaline phosphatase and acid phosphatase activities increased in shrimps kept at the lower salinities of 5, 15 and 35 g/L.

Vaseeharan B; Ramasamy P; Wesley SG; Chen JC

2013-06-01

147

Sodium alginate from Sargassum wightii retards mortalities in Penaeus monodon postlarvae challenged with white spot syndrome virus.  

UK PubMed Central (United Kingdom)

Sodium alginate extracted from brown seaweed Sargassum wightii (16.35 ± 1.42%, mean [±SD] yield from 5 extractions) was prepared as a powder or beads and used to enrich Artemia nauplii at concentrations of 100, 200, 300 and 400 mg l-1. The alginate-enriched nauplii were fed to Penaeus monodon shrimp postlarvae (PL) stage 15 (PL15, i.e. 15 d old) for 20 d. Mean weight gain and specific growth rate over this period were 0.24 g and 15.8%, respectively, in PL groups not fed alginate, and 0.20-0.28 g and 14.7-16.5%, respectively, in PL groups fed alginate. Amongst PL35 then challenged with white spot syndrome virus (WSSV) by immersion, all PL not fed alginate died within 9 d. However, amongst PL fed the 4 concentrations of alginate powder or beads, mortality rates reduced with increasing alginate concentration, and between 25 and 32% PL remained alive when the bioassay was terminated on Day 21. Amongst alginate-fed PL groups compared with the control group, mortality was reduced by 26.5 to 58.4%. Nested PCR detection of WSSV revealed sodium alginate concentration-dependent reductions in infection loads. The data indicate that sodium alginate extracted from brown seaweed and fed to P. monodon can retard progression of WSSV disease.

Immanuel G; Sivagnanavelmurugan M; Balasubramanian V; Palavesam A

2012-07-01

148

Sodium alginate from Sargassum wightii retards mortalities in Penaeus monodon postlarvae challenged with white spot syndrome virus.  

Science.gov (United States)

Sodium alginate extracted from brown seaweed Sargassum wightii (16.35 ± 1.42%, mean [±SD] yield from 5 extractions) was prepared as a powder or beads and used to enrich Artemia nauplii at concentrations of 100, 200, 300 and 400 mg l-1. The alginate-enriched nauplii were fed to Penaeus monodon shrimp postlarvae (PL) stage 15 (PL15, i.e. 15 d old) for 20 d. Mean weight gain and specific growth rate over this period were 0.24 g and 15.8%, respectively, in PL groups not fed alginate, and 0.20-0.28 g and 14.7-16.5%, respectively, in PL groups fed alginate. Amongst PL35 then challenged with white spot syndrome virus (WSSV) by immersion, all PL not fed alginate died within 9 d. However, amongst PL fed the 4 concentrations of alginate powder or beads, mortality rates reduced with increasing alginate concentration, and between 25 and 32% PL remained alive when the bioassay was terminated on Day 21. Amongst alginate-fed PL groups compared with the control group, mortality was reduced by 26.5 to 58.4%. Nested PCR detection of WSSV revealed sodium alginate concentration-dependent reductions in infection loads. The data indicate that sodium alginate extracted from brown seaweed and fed to P. monodon can retard progression of WSSV disease. PMID:22832717

Immanuel, Grasian; Sivagnanavelmurugan, Madasasmy; Balasubramanian, Varatharajan; Palavesam, Arunachalam

2012-07-25

149

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

Directory of Open Access Journals (Sweden)

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.

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

2011-01-01

150

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

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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.

X-Z Shi; X-C Li; S Wang; X-F Zhao; J-X Wang

2010-01-01

151

Prohibitin interacts with envelope proteins of white spot syndrome virus and prevents infection in red swamp crayfish Procambarus clarkii.  

UK PubMed Central (United Kingdom)

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 red swamp crayfish Procambarus clarkii, which was designated as 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 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 the controls. The effects of RNA silencing were rescued by rPcPHB1 re-injection. We further confirmed the interaction of PcPHB1 with the WSSV envelope proteins VP28, VP26, and VP24 using pull-down 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.

Lan JF; Li XC; Sun JJ; Gong J; Wang XW; Shi XZ; Shi LJ; Weng YD; Zhao XF; Wang JX

2013-09-01

152

Novel, closely related, white spot syndrome virus (WSSV) genotypes from Madagascar, Mozambique and the Kingdom of Saudi Arabia.  

Science.gov (United States)

White spot syndrome virus (WSSV) is highly pathogenic to penaeid shrimp and has caused significant economic losses in the aquaculture industry around the world. During 2010 to 2012, WSSV caused severe mortalities in cultured penaeid shrimp in Saudi Arabia, Mozambique and Madagascar. To investigate the origins of these WSSV, we performed genotyping analyses at 5 loci: the 3 open reading frames (ORFs) 125, 94 and 75, each containing a variable number of tandem repeats (VNTR), and deletions in the 2 variable regions, VR14/15 and VR23/24. We categorized the WSSV genotype as {N125, N94, N75, ?X14/15, ?X23/24} where N is the number of repeat units in a specific ORF and ?X is the length (base pair) of deletion within the variable region. We detected 4 WSSV genotypes, which were characterized by a full-length deletion in ORF94/95, a relatively small ORF75 and one specific deletion length in each variable region. There are 2 closely related genotypes in these 3 countries: {6125, del94, 375, ?595014/15, ?1097123/24} and {7125, del94, 375, ?595014/15, ?1097123/24}, where del is the full-length ORF deletion. In Saudi Arabia, 2 other related types of WSSV were also found: {6125, 794, 375, ?595014/15, ?1097123/24} and {8125, 1394, 375, ?595014/15, ?1097123/24}. The identical patterns of 3 loci in these 4 types indicate that they have a common lineage, and this suggests that the WSSV epidemics in these 3 countries were from a common source, possibly the environment. PMID:24062547

Tang, Kathy F J; Le Groumellec, Marc; Lightner, Donald V

2013-09-24

153

Novel, closely related, white spot syndrome virus (WSSV) genotypes from Madagascar, Mozambique and the Kingdom of Saudi Arabia.  

UK PubMed Central (United Kingdom)

White spot syndrome virus (WSSV) is highly pathogenic to penaeid shrimp and has caused significant economic losses in the aquaculture industry around the world. During 2010 to 2012, WSSV caused severe mortalities in cultured penaeid shrimp in Saudi Arabia, Mozambique and Madagascar. To investigate the origins of these WSSV, we performed genotyping analyses at 5 loci: the 3 open reading frames (ORFs) 125, 94 and 75, each containing a variable number of tandem repeats (VNTR), and deletions in the 2 variable regions, VR14/15 and VR23/24. We categorized the WSSV genotype as {N125, N94, N75, ?X14/15, ?X23/24} where N is the number of repeat units in a specific ORF and ?X is the length (base pair) of deletion within the variable region. We detected 4 WSSV genotypes, which were characterized by a full-length deletion in ORF94/95, a relatively small ORF75 and one specific deletion length in each variable region. There are 2 closely related genotypes in these 3 countries: {6125, del94, 375, ?595014/15, ?1097123/24} and {7125, del94, 375, ?595014/15, ?1097123/24}, where del is the full-length ORF deletion. In Saudi Arabia, 2 other related types of WSSV were also found: {6125, 794, 375, ?595014/15, ?1097123/24} and {8125, 1394, 375, ?595014/15, ?1097123/24}. The identical patterns of 3 loci in these 4 types indicate that they have a common lineage, and this suggests that the WSSV epidemics in these 3 countries were from a common source, possibly the environment.

Tang KF; Le Groumellec M; Lightner DV

2013-09-01

154

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

UK PubMed Central (United Kingdom)

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.

Jose S; Jayesh P; Sudheer NS; Poulose G; Mohandas A; Philip R; Singh IS

2012-05-01

155

Molecular docking analyses of Avicennia marina-derived phytochemicals against white spot syndrome virus (WSSV) envelope protein-VP28  

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Full Text Available White spot syndrome (WSS) is one of the most common and most disastrous diseases of shrimp worldwide. It causes up to 100% mortality within 3 to 4 days in commercial shrimp farms, resulting in large economic losses to the shrimp farming industry. VP28 envelope protein of WSSV is reported to play a key role in the systemic infection in shrimps. Considering the most sombre issue of viral disease in cultivated shrimp, the present study was undertaken to substantiate the inhibition potential of Avicennia marina-derived phytochemicals against the WSSV envelope protein VP28. Seven A. marina-derived phytochemicals namely stigmasterol, triterpenoid, betulin, lupeol, avicenol-A, betulinic acid and quercetin were docked against the WSSV protein VP28 by using Argus lab molecular docking software. The chemical structures of the phytochemicals were retrieved from Pubchem database and generated from SMILES notation. Similarly the protein structure of the envelope protein was obtained from protein data bank (PDB-ID: 2ED6). Binding sites were predicted by using ligand explorer software. Among the phytochemicals screened, stigmasterol, lupeol and betulin showed the best binding exhibiting the potential to block VP28 envelope protein of WSSV, which could possibly inhibit the attachment of WSSV to the host species. Further experimental studies will provide a clear understanding on the mode of action of these phytochemicals individually or synergistically against WSSV envelope protein and can be used as an inhibitory drug to reduce white spot related severe complications in crustaceans.

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

2012-01-01

156

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

Science.gov (United States)

White spot syndrome (WSS) is one of the most common and most disastrous diseases of shrimp worldwide. It causes up to 100% mortality within 3 to 4 days in commercial shrimp farms, resulting in large economic losses to the shrimp farming industry. VP28 envelope protein of WSSV is reported to play a key role in the systemic infection in shrimps. Considering the most sombre issue of viral disease in cultivated shrimp, the present study was undertaken to substantiate the inhibition potential of Avicennia marinaderived phytochemicals against the WSSV envelope protein VP28. Seven A. marina-derived phytochemicals namely stigmasterol, triterpenoid, betulin, lupeol, avicenol-A, betulinic acid and quercetin were docked against the WSSV protein VP28 by using Argus lab molecular docking software. The chemical structures of the phytochemicals were retrieved from Pubchem database and generated from SMILES notation. Similarly the protein structure of the envelope protein was obtained from protein data bank (PDB-ID: 2ED6). Binding sites were predicted by using ligand explorer software. Among the phytochemicals screened, stigmasterol, lupeol and betulin showed the best binding exhibiting the potential to block VP28 envelope protein of WSSV, which could possibly inhibit the attachment of WSSV to the host species. Further experimental studies will provide a clear understanding on the mode of action of these phytochemicals individually or synergistically against WSSV envelope protein and can be used as an inhibitory drug to reduce white spot related severe complications in crustaceans.

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

2012-01-01

157

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

UK PubMed Central (United Kingdom)

White spot syndrome (WSS) is one of the most common and most disastrous diseases of shrimp worldwide. It causes up to 100% mortality within 3 to 4 days in commercial shrimp farms, resulting in large economic losses to the shrimp farming industry. VP28 envelope protein of WSSV is reported to play a key role in the systemic infection in shrimps. Considering the most sombre issue of viral disease in cultivated shrimp, the present study was undertaken to substantiate the inhibition potential of Avicennia marinaderived phytochemicals against the WSSV envelope protein VP28. Seven A. marina-derived phytochemicals namely stigmasterol, triterpenoid, betulin, lupeol, avicenol-A, betulinic acid and quercetin were docked against the WSSV protein VP28 by using Argus lab molecular docking software. The chemical structures of the phytochemicals were retrieved from Pubchem database and generated from SMILES notation. Similarly the protein structure of the envelope protein was obtained from protein data bank (PDB-ID: 2ED6). Binding sites were predicted by using ligand explorer software. Among the phytochemicals screened, stigmasterol, lupeol and betulin showed the best binding exhibiting the potential to block VP28 envelope protein of WSSV, which could possibly inhibit the attachment of WSSV to the host species. Further experimental studies will provide a clear understanding on the mode of action of these phytochemicals individually or synergistically against WSSV envelope protein and can be used as an inhibitory drug to reduce white spot related severe complications in crustaceans.

Sahu SK; Kathiresan K; Singh R; Senthilraja P

2012-01-01

158

Saturn's Great White Spots  

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The history of observations of the Great White Spots on Saturn is reviewed, incluing the 1876, 1903, 1933, and 1960 outbreaks. The spots have been used to trace zonal winds on Saturn, showing longitudinal expansion spot velocities of 30 to 45 m/s and north-south expansion velocities of about 3 m/s. The relationship between spot activity and the Saturnian atmosphere is discussed. A graph of insolation at the top of Saturn's atmosphere during Great White Spot activity and photographs of the 1933 and 1960 spots are presented. Because the trends in spot activity show a recurrence time of about one Saturnian year (29.51 yrs), it is suggested that spot activity may occur in 1989 or 1990.

Sanchez-Lavega, A. (Universidad del Pais Vasco, Bilbao (Spain))

1989-08-01

159

The C-terminal region of envelope protein VP38 from white spot syndrome virus is indispensable for interaction with VP24.  

UK PubMed Central (United Kingdom)

White spot syndrome virus (WSSV) is a large, rod-shaped, enveloped double-stranded DNA virus. In this study, VP38, a viral envelope protein, was expressed as a glutathione S-transferase (GST) fusion protein, and a polyclonal antibody against VP38 was obtained. Far-Western blotting and GST pull-down showed that VP38 interacted directly with VP24, a major WSSV envelope protein. In addition, to delineate the interaction region of VP38 with VP24, GST-VP38n (aa 1-142) and GST-VP38c (aa 143-309) were expressed. The GST pull-down assay revealed that VP38 binds via its C-terminal region to VP24. The result implies that VP38 may participate in the formation of the WSSV envelope.

Jie Z; Xu L; Yang F

2008-01-01

160

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

Science.gov (United States)

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 ?g 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. PMID:23617699

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

2013-04-26

 
 
 
 
161

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

UK PubMed Central (United Kingdom)

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.

Ponprateep S; Phiwsaiya K; Tassanakajon A; Rimphanitchayakit V

2013-09-01

162

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

Science.gov (United States)

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. PMID:23867494

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

2013-07-16

163

Antiviral activity of bis(2-methylheptyl)phthalate isolated from Pongamia pinnata leaves against White Spot Syndrome Virus of Penaeus monodon Fabricius.  

UK PubMed Central (United Kingdom)

White Spot Syndrome Virus (WSSV) is an extremely virulent, contagious, causative agent of the White spot syndrome of shrimp and causes high mortality and affects most of the commercially important cultured marine crustacean species globally. Oral administration of ethanolic extract and purified compound from the leaves of Pongamia pinnata, an indigenious Indian "medicinal plant" "has increased the survival of WSSV infected Penaeus monodon". Pelletized feed impregnated with ethanolic extract of the leaves of P. pinnata was fed to shrimp prior and after WSSV infection at 200 and 300 microg/g of body weight of shrimp/day. The survival rate for the WSSV-infected shrimp that were fed with 200 and 300 microg extract/g were 40% and 80%, respectively. The active WSSV antiviral compound 1 that was isolated from the leaves of P. pinnata was identified as bis(2-methylheptyl)phthalate. Thus, the present work revealed that oral administration of the crude and purified compound from the leaves of P. pinnata effectively inhibited WSSV pathogenesis and reduced the mortality of infected shrimp.

Rameshthangam P; Ramasamy P

2007-06-01

164

Identification of RAPD-SCAR marker linked to white spot syndrome virus resistance in populations of giant black tiger shrimp, Penaeus monodon Fabricius.  

UK PubMed Central (United Kingdom)

White spot disease (WSD) caused by white spot syndrome virus (WSSV) creates severe epizootics in shrimp aquaculture industry worldwide. Despite several efforts, no such permanent remedy was yet developed. Selective breeding using DNA markers would be a cost-effective strategy for long-term solution of this problem. In the present investigation, out of 30 random primers, only one primer produced a statistically significant (P < 0.01) randomly amplified polymorphic DNA (RAPD) marker of 502 bp, which provided a good discrimination between disease resistant and disease susceptible populations of Penaeus monodon from three geographical locations along the East coast of India. Because RAPD markers are dominant, a sequence characterized amplified region (SCAR) marker was developed by cloning and sequencing of 502 bp RAPD fragment, which generates a single 457 bp DNA fragment after PCR amplification only in the disease resistant shrimps. Challenge experiment was also conducted to validate this 457 bp SCAR marker, and the results suggested that the WSSV loads were 2.25 × 10(3) fold higher in disease susceptible than that in disease resistant shrimps using real-time PCR. Therefore, this 457 bp DNA SCAR marker will be very valuable towards the development of disease-free shrimp aquaculture industry.

Dutta S; Biswas S; Mukherjee K; Chakrabarty U; Mallik A; Mandal N

2013-08-01

165

Evidence for the presence of white spot syndrome virus (WSSV) and monodon baculovirus (MBV) in wild Penaeus monodon (Fabricius) broodstock, in the southeast coast of India.  

Science.gov (United States)

A survey on the presence of the viruses of two economically significant diseases, white spot syndrome virus (WSSV) and monodon baculovirus (MBV) in wild-collected Penaeus monodon broodstock, was conducted during different seasons of the year in two major coastal areas of southeast India. The broodstock were collected along the coast of Tamil Nadu and Andhra Pradesh during summer, premonsoon, monsoon and post-monsoon seasons for three consecutive years. A total of 7905 samples were collected and subjected to MBV screening, and 6709 samples that were screened as MBV negative were diagnosed for WSSV. MBV was detected using rapid malachite green staining and WSSV by nested polymerase chain reaction. Prevalence data of the viruses were analysed using the EpiCalc 2000 program at 95% confidence interval. Samples collected from the Andhra Pradesh coast displayed a slightly higher prevalence of WSSV and MBV infection than those collected from Tamil Nadu, although this difference was not statistically significant (P > 005). In addition, it was found that the prevalence of both WSSV and MBV infections fluctuated according to season. Data on prevalence of these viruses in broodstock would be useful to develop strategies for shrimp health management along the southeast coast of India. PMID:22924635

Remany, M C; Daly, C; Nagaraj, S; Panda, A K; Jaideep, K; Samraj, Y C T

2012-08-23

166

A new method for quantifying white spot syndrome virus: Experimental challenge dose using TaqMan real-time PCR assay.  

UK PubMed Central (United Kingdom)

White spot syndrome virus (WSSV) is an important pathogen in shrimp aquaculture. The susceptibility of crayfish (Procambarus clarkii) was assessed by means of serial dilutions of a solution containing WSSV. A TaqMan real-time PCR was used to quantify the WSSV challenge dose in P. clarkii. The results showed that WSSV copies could be detected at concentrations from 1.365×10(4) to 1.129×10(9) copies/?l. The viral infectivity (LD(50)), measured as the mortality of infected crayfish, indicated 60% mortality in the 10(5) dilution group (1.524×10(5) copies/?l). TaqMan real-time PCR represents a novel standard method, based on the by quantitation of WSSV copies, for determining the appropriate concentration of WSSV for use in infection experiments.

Zhu F; Quan H

2012-09-01

167

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

Directory of Open Access Journals (Sweden)

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.

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

2011-01-01

168

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

Scientific Electronic Library Online (English)

Full Text Available Abstract in english 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 applicatio (more) ns, 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.

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

2011-04-01

169

Identification of white spot syndrome virus latency-related genes in specific-pathogen-free shrimps by use of a microarray.  

UK PubMed Central (United Kingdom)

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

Khadijah S; Neo SY; Hossain MS; Miller LD; Mathavan S; Kwang J

2003-09-01

170

The Unique Stacked Rings in the Nucleocapsid of the White Spot Syndrome Virus Virion Are Formed by the Major Structural Protein VP664, the Largest Viral Structural Protein Ever Found  

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One unique feature of the shrimp white spot syndrome virus (WSSV) genome is the presence of a giant open reading frame (ORF) of 18,234 nucleotides that encodes a long polypeptide of 6,077 amino acids with a hitherto unknown function. In the present study, by applying proteomic methodology to analyze...

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

171

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

Science.gov (United States)

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. PMID:23097446

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

2012-10-24

172

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

UK PubMed Central (United Kingdom)

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.

Chen AJ; Gao L; Wang XW; Zhao XF; Wang JX

2013-01-01

173

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

Science.gov (United States)

We analysed 20 boxes of, frozen imported bait-shrimp (China: Parapenaeopsis sp. and Metapenaeopsis sp.) and 8 boxes of native, frozen bait-shrimp (Gulf of Mexico: Litopenaeus setiferus and Farfantepenaeus duorarum) by RT-PCR or PCR for Taura syndrome virus (TSV), yellowhead virus/gill-associated virus (YHV/GAV), white-spot syndrome virus (WSSV) and infectious hypodermal and hematopoietic necrosis virus (IHHNV). All 28 boxes of shrimp were negative for TSV, YHV/GAV and IHHNV; 2 boxes of imported bait-shrimp were WSSV-positive by 3 different PCR assays. Intramuscular injection of replicate groups of SPF (specific pathogen-free) L. vannamei juveniles with 2 different tissue homogenates prepared from the 2 WSSV-positive bait boxes resulted in 100% mortality of the test shrimp within 48 to 72 h post-injection. No mortality occurred among injected negative control groups. Histological and in situ hybridization analyses of 20 moribund treatment-shrimp demonstrated severe WSSV infections in each sample. Oral exposure of SPF L. vannamei postlarvae, PL (PL 25 to 30 stage; approximately 0.02 g) to minced tissue prepared from the 2 WSSV-positive bait-lots did not induce infection, possibly because of an insufficient infectious dose and/or viral inactivation resulting from multiple freeze-thaw cycles of the bait-shrimp during PCR testing. Use of an electric drill and collection of drill-tailings (tissue from approximately 20 to 30 shrimp) from frozen blocks of shrimp was successfully employed as an alternate tissue-sampling method without thawing. Our findings indicate that imported WSSV-infected bait shrimp, originating from China, are being sold in Texas for the purpose of sport fishing and represent a potential threat to freshwater and marine crustacean fisheries, as well as to coastal US shrimp farms. PMID:16956056

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

2006-07-25

174

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

UK PubMed Central (United Kingdom)

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

Hasson KW; Fan Y; Reisinger T; Venuti J; Varner PW

2006-07-01

175

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

UK PubMed Central (United Kingdom)

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 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 activated E2F1 to stimulate G1/S transition. Our findings provide the first evidence that WSSV may regulate cell cycle progression by targeting the Rb-E2F pathway.

Ran X; Bian X; Ji Y; Yan X; Yang F; Li F

2013-09-01

176

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

Science.gov (United States)

White spot syndrome virus (WSSV) is the most important pathogen in shrimp farming systems worldwide including the Mekong Delta, Vietnam. The genome of WSSV is characterized by the presence of two major 'indel regions' found at ORF14/15 and ORF23/24 (WSSV-Thailand) and three regions with variable number tandem repeats (VNTR) located in ORF75, ORF94 and ORF125. In the current study, we investigated whether or not the number of repeat units in the VNTRs correlates with virus outbreak status and/or shrimp farming practice. We analysed 662 WSSV samples from individual WSSV-infected Penaeus monodon shrimp from 104 ponds collected from two important shrimp farming regions of the Mekong Delta: Ca Mau and Bac Lieu. Using this large data set and statistical analysis, we found that for ORF94 and ORF125, the mean number of repeat units (RUs) in VNTRs was significantly lower in disease outbreak ponds than in non-outbreak ponds. Although a higher mean RU number was observed in the improved-extensive system than in the rice-shrimp or semi-intensive systems, these differences were not significant. VNTR sequences are thus not only useful markers for studying WSSV genotypes and populations, but specific VNTR variants also correlate with disease outbreaks in shrimp farming systems. PMID:22913744

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

2012-08-23

177

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

UK PubMed Central (United Kingdom)

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 2,003 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.

Yingvilasprasert W; Supungul P; Tassanakajon A

2013-09-01

178

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

Science.gov (United States)

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.

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

2004-10-01

179

White-centred retinal haemorrhages (Roth spots).  

UK PubMed Central (United Kingdom)

Roth spots (white-centred retinal haemorrhages) were classically described as septic emboli lodged in the retina of patients with subacute bacterial endocarditis. Indeed many have considered Roth spots pathognomonic for this condition. More recent histological evidence suggests, however, that they are not foci of bacterial abscess. Instead, they are nonspecific and may be found in many other diseases. A review of the histology and the pathogenesis of these white-centred haemorrhages will be provided, along with the work-up of the differential diagnosis.

Ling R; James B

1998-10-01

180

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

Energy Technology Data Exchange (ETDEWEB)

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

Tang,X.; Wu, J.; Sivaraman, J.; Hew, C.

2007-01-01

 
 
 
 
181

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

Energy Technology Data Exchange (ETDEWEB)

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

Liu,Y.; Wu, J.; Song, J.; Sivaraman, J.; Hew, C.

2006-01-01

182

Interaction between shrimp and white spot syndrome virus through PmRab7-VP28 complex: an insight using simulation and docking studies.  

Science.gov (United States)

White spot disease is a devastating disease of shrimp Penaeus monodon in which the shrimp receptor protein PmRab7 interacts with viral envelop protein VP28 to form PmRab7-VP28 complex, which causes initiation of the disease. The molecular mechanism implicated in the disease, the dynamic behavior of proteins as well as interaction between both the biological counterparts that crafts a micro-environment feasible for entry of virus into the shrimp is still unknown. In the present study, we applied molecular modeling (MM), molecular dynamics (MD) and docking to compute surface mapping of infective amino acid residues between interacting proteins. Our result showed that ?-helix of PmRab7 (encompassing Ser74, Ile143, Thr184, Arg53, Asn144, Thr184, Arg53, Arg79) interacts with ?-sheets of VP28 (containing Ser74, Ile143, Thr184, Arg53, Asn144, Thr184, Arg53, Arg79) and Arg69-Ser74, Val75-Ile143, Leu73-Ile143, Arg79-Asn144, Ala198-Ala182 bonds contributed in the formation of PmRab7-VP28 complex. Further studies on the amino acid residues and bonds may open new possibilities for preventing PmRab7-VP28 complex formation, thus reducing chances of WSD. The quantitative predictions provide a scope for experimental testing in future as well as endow with a straightforward evidence to comprehend cellular mechanisms underlying the disease. PMID:23179770

Verma, Arunima Kumar; Gupta, Shipra; Verma, Sharad; Mishra, Abha; Nagpure, N S; Singh, Shivesh Pratap; Pathak, Ajey Kumar; Sarkar, Uttam Kumar; Singh, Shri Prakash; Singh, Mahender; Seth, Prahlad Kishore

2012-11-23

183

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

UK PubMed Central (United Kingdom)

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

Zhu F; Zhang X

2012-02-01

184

A Kazal type serine proteinase SPIPm2 from the black tiger shrimp Penaeus monodon is capable of neutralization and protection of hemocytes from the white spot syndrome virus.  

UK PubMed Central (United Kingdom)

A Kazal type serine proteinase SPIPm2 is abundantly expressed in the hemocytes and shown to be involved in innate immune response against white spot syndrome virus (WSSV) in Penaeus monodon. The SPIPm2 is expressed and stored in the granules in the cytoplasm of semigranular and granular but not the hyaline hemocytes. Upon WSSV challenge and progression of infection, the SPIPm2 was secreted readily from the semigranular and granular hemocytes. The more they secreted the SPIPm2, the less they were distinguishable from the hyaline cells. The WSSV-infected cells were either semigranular or granular hemocytes or both and depleted of SPIPm2. The rSPIPm2 was able to temporarily and dose-dependently neutralize the WSSV and protect the hemocytes from viral infection judging from the substantially less expression of WSSV late gene VP28. The antiviral activity was very likely due to the binding of SPIPm2 to the components of viral particle and hemocyte cell membrane.

Ponprateep S; Tassanakajon A; Rimphanitchayakit V

2011-12-01

185

A Kazal type serine proteinase SPIPm2 from the black tiger shrimp Penaeus monodon is capable of neutralization and protection of hemocytes from the white spot syndrome virus.  

Science.gov (United States)

A Kazal type serine proteinase SPIPm2 is abundantly expressed in the hemocytes and shown to be involved in innate immune response against white spot syndrome virus (WSSV) in Penaeus monodon. The SPIPm2 is expressed and stored in the granules in the cytoplasm of semigranular and granular but not the hyaline hemocytes. Upon WSSV challenge and progression of infection, the SPIPm2 was secreted readily from the semigranular and granular hemocytes. The more they secreted the SPIPm2, the less they were distinguishable from the hyaline cells. The WSSV-infected cells were either semigranular or granular hemocytes or both and depleted of SPIPm2. The rSPIPm2 was able to temporarily and dose-dependently neutralize the WSSV and protect the hemocytes from viral infection judging from the substantially less expression of WSSV late gene VP28. The antiviral activity was very likely due to the binding of SPIPm2 to the components of viral particle and hemocyte cell membrane. PMID:22032902

Ponprateep, Sirikwan; Tassanakajon, Anchalee; Rimphanitchayakit, Vichien

2011-10-20

186

Detection of two distinct types of hemolymphatic prophenoloxidase and their differential responses in the black tiger shrimp, Penaeus monodon, upon infection by white spot syndrome virus  

UK PubMed Central (United Kingdom)

The present study demonstrates the existence of two distinct types of prophenoloxidases in the serum of the black tiger shrimp, Penaeus monodon, and their differential responses upon infection by white spot syndrome virus (WSSV). Empirical analyses performed using various activators (proteases and detergents) and phenolic substrates revealed that the anionic detergent, sodium dodecyl sulfate, was the best exogenous activator of serum prophenoloxidase (designated as prophenoloxidase-1), and the resulting phenoloxidase maximally oxidized an o-diphenolic substrate, dopamine (K?: 0.90mM). A novel type of serum prophenoloxidase (designated as prophenoloxidase-2), with an intrinsic property of spontaneous conversion to phenoloxidase, was also detected, for the first time, in healthy shrimps. Interestingly, this phenoloxidase exhibited profound affinity for a monophenolic substrate tyramine (K?: 1.25mM), and the level of prophenoloxidase-2 was 2.4-fold higher than that of prophenoloxidase-1. Strikingly, WSSV infection in shrimps inflicted a remarkable elevation in the level of prophenoloxidase-1 and concomitant depletion of prophenoloxidase-2, thereby demonstrating differential responses of these two distinct types of serum prophenoloxidases. The findings of this investigation implicate an important role of prophenoloxidase-2 in conferring antiviral state to shrimps.

Radha S; Mullainadhan P; Arumugam M

2013-02-01

187

The Marsupenaeus japonicus voltage-dependent anion channel (MjVDAC) protein is involved in white spot syndrome virus (WSSV) pathogenesis.  

Science.gov (United States)

Voltage-dependent anion channel (VDAC) proteins abound in the outer membrane of mitochondria. They play an important role in mitochondrial membrane permeabilization (MMP), which can lead to stress-induced cellular apoptosis and necrosis. Several pathogens regulate this MMP in their host cells to benefit their replication cycle, while in other cases, the host can use the same mechanism to combat pathogenesis. In this study, the first shrimp VDAC gene was identified and characterized from Marsupenaeus japonicus (MjVDAC). Its open reading frame (ORF) contained 849 bp encoding 282 amino acids. The deduced MjVDAC protein includes the 4-element eukaryotic porin signature motif, the conserved ATP binding motif (the GLK motif) and a VKAKV-like sequence known in other organisms to be involved in the protein's incorporation in the mitochondrial outer membrane. Tissue tropism analysis indicated that MjVDAC is abundant in the heart, muscle, stomach and pleopod. MjVDAC proteins colocalized with mitochondria in transiently transfected Sf9 cells and in shrimp hemocytes. dsRNA silencing of shrimp VDAC delayed white spot syndrome virus (WSSV) infection by 1 day in different shrimp organs. Taken together, these findings suggest that MjVDAC is likely to be involved in WSSV pathogenesis. PMID:20202479

Wang, K C Han-Ching; Kondo, Hidehiro; Hirono, Ikuo; Aoki, Takashi

2010-03-01

188

The Marsupenaeus japonicus voltage-dependent anion channel (MjVDAC) protein is involved in white spot syndrome virus (WSSV) pathogenesis.  

UK PubMed Central (United Kingdom)

Voltage-dependent anion channel (VDAC) proteins abound in the outer membrane of mitochondria. They play an important role in mitochondrial membrane permeabilization (MMP), which can lead to stress-induced cellular apoptosis and necrosis. Several pathogens regulate this MMP in their host cells to benefit their replication cycle, while in other cases, the host can use the same mechanism to combat pathogenesis. In this study, the first shrimp VDAC gene was identified and characterized from Marsupenaeus japonicus (MjVDAC). Its open reading frame (ORF) contained 849 bp encoding 282 amino acids. The deduced MjVDAC protein includes the 4-element eukaryotic porin signature motif, the conserved ATP binding motif (the GLK motif) and a VKAKV-like sequence known in other organisms to be involved in the protein's incorporation in the mitochondrial outer membrane. Tissue tropism analysis indicated that MjVDAC is abundant in the heart, muscle, stomach and pleopod. MjVDAC proteins colocalized with mitochondria in transiently transfected Sf9 cells and in shrimp hemocytes. dsRNA silencing of shrimp VDAC delayed white spot syndrome virus (WSSV) infection by 1 day in different shrimp organs. Taken together, these findings suggest that MjVDAC is likely to be involved in WSSV pathogenesis.

Wang KC; Kondo H; Hirono I; Aoki T

2010-07-01

189

Effect of hot water extracts of brown seaweeds Sargassum spp. on growth and resistance to white spot syndrome virus in shrimp Penaeus monodon postlarvae  

UK PubMed Central (United Kingdom)

An experiment was conducted to evaluate the effect of a hot water extract of brown seaweeds Sargassum duplicatum and Sargassum wightii on the growth and white spot syndrome virus (WSSV) resistance in shrimp Penaeus monodon postlarvae (PL). Artemia nauplii (instar II) were enriched with both seaweed extracts at various concentrations (250, 500 and 750 mg L?¹) and fed to the respective P. monodon (PL15-35) group for 20 days. A control group was also maintained without seaweed extract supplementation. The weight gain of the experimental groups was significantly higher (0.274-0.323 g) than the control group (0.261 g). Similarly, the specific growth rate was also significantly higher (16.27-17.06%) in the experimental groups than in the control group (16.03%). After 20 days of the feeding experiment, the shrimp PL were challenged with WSSV for 21 days. During the challenge test, the control shrimp displayed 100% mortality within 8 days. In contrast, the mortality percentage of the highest concentration (750 mg L?¹) of seaweed extract enriched Artemia nauplii fed shrimp was 54-79%. Comparatively, low mortality was observed in S. wightii extract-enriched Artemia nauplii fed shrimp. The polymerase chain reaction analysis indicated the concentration-dependent infection of WSSV in P. monodon PL.

Immanuel G; Sivagnanavelmurugan M; Balasubramanian V; Palavesam A

2010-09-01

190

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

Science.gov (United States)

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

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

2013-01-01

191

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

UK PubMed Central (United Kingdom)

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

Wang PH; Gu ZH; Wan DH; Liu BD; Huang XD; Weng SP; Yu XQ; He JG

2013-09-01

192

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

Science.gov (United States)

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

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

2013-08-19

193

Development of a multiplex loop-mediated isothermal amplification (mLAMP) method for the simultaneous detection of white spot syndrome virus and infectious hypodermal and hematopoietic necrosis virus in penaeid shrimp  

UK PubMed Central (United Kingdom)

A multiplex LAMP (mLAMP) assay was developed for the simultaneous detection of two penaeid shrimp viruses, namely, white spot syndrome virus (WSSV) and infectious hypodermal and hematopoietic necrosis virus (IHHNV), which are the major viral pathogens of penaeid shrimp worldwide. Two sets of the LAMP primers were designed for VP28 gene of WSSV and non-structural protein 1 (NS1) gene of IHHNV, in which a restriction enzyme cleavage site was inserted into two pairs of species-specific primers, to construct the mLAMP assay by combining these two sets totaling eight primers. The mLAMP method was distinguishable between WSSV and IHHNV due to the subsequent restriction enzyme analysis. The sensitivities of the mLAMP method were 10² and 10³ times higher on the detection limits for WSSV and IHHNV, respectively, than those of the nested-PCR (nPCR) and classical PCR methods. No positive results were displayed when nucleic acids from other shrimp pathogen DNAs were used as mLAMP templates. Uracil-N-glycosylase (UNG) was used to prevent carry-over contamination of mLAMP products. In addition to its high specificity and sensitivity, the developed multiplex LAMP assay offers an efficient, convenient and rapid tool for screening penaeid shrimp viruses since both WSSV and IHHNV can be diagnosed in a single reaction, and will be useful for the control of these viruses in shrimp.

He L; Xu Hs

2011-02-01

194

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

Scientific Electronic Library Online (English)

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

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

2012-06-01

195

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

Directory of Open Access Journals (Sweden)

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

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

2012-01-01

196

Shrimp Dscam and its cytoplasmic tail splicing activator serine/arginine (SR)-rich protein B52 were both induced after white spot syndrome virus challenge.  

Science.gov (United States)

The serine/arginine (SR)-rich protein family is phylogenetically conserved and plays significant roles in mRNA maturation, including alternative splicing (AS). In Drosophila, SR protein B52 functions as a splicing activator to regulate AS events in several genes, including the Down syndrome cell adhesion molecule (Dscam). In this study, the B52 gene from Litopenaeus vannamei (LvB52) was isolated and characterized. The open reading frame of LvB52 contains 1149 bp encoding 382 amino acids. The deduced LvB52 protein includes two RNA recognition motifs (RRM) at the N terminus and an arginine/serine rich domain (RS rich domain) at the C terminus, and thus shows the expected RRM1-RRM2-RS domain architecture. Tissue tropism analysis revealed that LvB52 is expressed in most tissues and at high levels in stomach and muscle. After white spot syndrome virus (WSSV) infection, a parallel increase in the expression of total LvDscam, tail-less LvDscam, membrane-bound LvDscam and LvB52 was observed after 24 hpi. Conversely, there was no obvious change in the expression of the AS repressor Lvhrp36. In vivo dsRNA silencing of LvB52 induced element 3 exclusion in the LvDscam cytoplasmic tail, but no abnormal exclusions in the Ig2-Ig3 region or the transmembrane region. We also found that the exon of the Ig7 region was quite often excluded, even in normal shrimp, and that LvB52 silencing was associated with a decrease in the variability of this region. Taken together, our data suggest that LvB52 acts as a splicing activator that regulates AS events in LvDscam. PMID:23123640

Chiang, Yi-An; Hung, Hsin-Yi; Lee, Chung-Wei; Huang, Yun-Tzu; Wang, Han-Ching

2012-10-30

197

Shrimp Dscam and its cytoplasmic tail splicing activator serine/arginine (SR)-rich protein B52 were both induced after white spot syndrome virus challenge.  

UK PubMed Central (United Kingdom)

The serine/arginine (SR)-rich protein family is phylogenetically conserved and plays significant roles in mRNA maturation, including alternative splicing (AS). In Drosophila, SR protein B52 functions as a splicing activator to regulate AS events in several genes, including the Down syndrome cell adhesion molecule (Dscam). In this study, the B52 gene from Litopenaeus vannamei (LvB52) was isolated and characterized. The open reading frame of LvB52 contains 1149 bp encoding 382 amino acids. The deduced LvB52 protein includes two RNA recognition motifs (RRM) at the N terminus and an arginine/serine rich domain (RS rich domain) at the C terminus, and thus shows the expected RRM1-RRM2-RS domain architecture. Tissue tropism analysis revealed that LvB52 is expressed in most tissues and at high levels in stomach and muscle. After white spot syndrome virus (WSSV) infection, a parallel increase in the expression of total LvDscam, tail-less LvDscam, membrane-bound LvDscam and LvB52 was observed after 24 hpi. Conversely, there was no obvious change in the expression of the AS repressor Lvhrp36. In vivo dsRNA silencing of LvB52 induced element 3 exclusion in the LvDscam cytoplasmic tail, but no abnormal exclusions in the Ig2-Ig3 region or the transmembrane region. We also found that the exon of the Ig7 region was quite often excluded, even in normal shrimp, and that LvB52 silencing was associated with a decrease in the variability of this region. Taken together, our data suggest that LvB52 acts as a splicing activator that regulates AS events in LvDscam.

Chiang YA; Hung HY; Lee CW; Huang YT; Wang HC

2013-01-01

198

Activating transcription factor 4 and X box binding protein 1 of Litopenaeus vannamei transcriptional regulated white spot syndrome virus genes Wsv023 and Wsv083.  

UK PubMed Central (United Kingdom)

In response to endoplasmic reticulum (ER) stress, the signaling pathway termed unfolded protein response (UPR) is activated. To investigate the role of UPR in Litopenaeus vannamei immunity, the activating transcription factor 4 (designated as LvATF4) which belonged to a branch of the UPR, the [protein kinase RNA (PKR)-like ER kinase, (PERK)]-[eukaryotic initiation factor 2 subunit alpha (eIF2?)] pathway, was identified and characterized. The full-length cDNA of LvATF4 was 1972 bp long, with an open reading frame of 1299 bp long that encoded a 432 amino acid protein. LvATF4 was highly expressed in gills, intestines and stomach. For the white spot syndrome virus (WSSV) challenge, LvATF4 was upregulated in the gills after 3 hpi and increased by 1.9-fold (96 hpi) compared to the mock-treated group. The LvATF4 knock-down by RNA interference resulted in a lower cumulative mortality of L. vannamei under WSSV infection. Reporter gene assays show that LvATF4 could upregulate the expression of the WSSV gene wsv023 based on the activating transcription factor/cyclic adenosine 3', 5'-monophosphate response element (ATF/CRE). Another transcription factor of L. vannamei, X box binding protein 1 (designated as LvXBP1), has a significant function in [inositol-requiring enzyme-1(IRE1) - (XBP1)] pathway. This transcription factor upregulated the expression of the WSSV gene wsv083 based on the UPR element (UPRE). These results suggest that in L. vannamei UPR signaling pathway transcription factors are important for WSSV and might facilitate WSSV infection.

Li XY; Pang LR; Chen YG; Weng SP; Yue HT; Zhang ZZ; Chen YH; He JG

2013-01-01

199

Activating transcription factor 4 and X box binding protein 1 of Litopenaeus vannamei transcriptional regulated white spot syndrome virus genes Wsv023 and Wsv083.  

Science.gov (United States)

In response to endoplasmic reticulum (ER) stress, the signaling pathway termed unfolded protein response (UPR) is activated. To investigate the role of UPR in Litopenaeus vannamei immunity, the activating transcription factor 4 (designated as LvATF4) which belonged to a branch of the UPR, the [protein kinase RNA (PKR)-like ER kinase, (PERK)]-[eukaryotic initiation factor 2 subunit alpha (eIF2?)] pathway, was identified and characterized. The full-length cDNA of LvATF4 was 1972 bp long, with an open reading frame of 1299 bp long that encoded a 432 amino acid protein. LvATF4 was highly expressed in gills, intestines and stomach. For the white spot syndrome virus (WSSV) challenge, LvATF4 was upregulated in the gills after 3 hpi and increased by 1.9-fold (96 hpi) compared to the mock-treated group. The LvATF4 knock-down by RNA interference resulted in a lower cumulative mortality of L. vannamei under WSSV infection. Reporter gene assays show that LvATF4 could upregulate the expression of the WSSV gene wsv023 based on the activating transcription factor/cyclic adenosine 3', 5'-monophosphate response element (ATF/CRE). Another transcription factor of L. vannamei, X box binding protein 1 (designated as LvXBP1), has a significant function in [inositol-requiring enzyme-1(IRE1) - (XBP1)] pathway. This transcription factor upregulated the expression of the WSSV gene wsv083 based on the UPR element (UPRE). These results suggest that in L. vannamei UPR signaling pathway transcription factors are important for WSSV and might facilitate WSSV infection. PMID:23638122

Li, Xiao-Yun; Pang, Li-Ran; Chen, Yong-Gui; Weng, Shao-Ping; Yue, Hai-Tao; Zhang, Ze-Zhi; Chen, Yi-Hong; He, Jian-Guo

2013-04-24

200

Influence of acute salinity changes on biochemical, hematological and immune indices of Fenneropenaeus indicus during white spot syndrome virus (WSSV) challenges.  

UK PubMed Central (United Kingdom)

The present study reports the influence of salinity (5, 15, 25 and 35?g/L) on the biochemical and immune parameters of Fenneropenaeus indicus challenged with 5. 5?×?10(4) copy number of White Spot Syndrome Virus. F. indicus that were reared in 25?g/L, were injected with WSSV, and then transferred to 5, 15, 25 (control) and 35g/L were examined for total haemocyte count (THC), phenoloxidase (PO) activity, superoxide dismutase (SOD) activity, alkaline phosphatase assay and acid phosphatase assay after 0?120?h. It was concluded that the F. indicus transferred from 25?g/L to low and higher salinity levels (5?g/L, 15?g/L and 35?g/L) had reduced immune indices and decreased resistance against WSSV infection. After 120?h, the mortality rate in WSSV injected F. indicus experimental groups (5?g/L and 35?g/L) was significantly higher than in F. indicus exposed to 25?g/L and 15?g/L salinities. During the experimental period (0-120?h), biochemical variables viz. total protein, total carbohydrate, and total lipid levels were measured in the hemolymph of both experimental and control groups. Acute salinity changes induced an increase in protein variations across the salinity ranges in shrimps. The THC, and PO activity, decreased significantly while SOD activity, alkaline phosphatase activity, and acid phosphatase activity was increased in the shrimp held at lower salinity of 5?g/L, 15?g/L and 35?g/L after 24?h.

Vaseeharan B; Ramasamy P; Wesley G; Chen JC

2013-04-01

 
 
 
 
201

Influence of acute salinity changes on biochemical, hematological and immune indices of Fenneropenaeus indicus during white spot syndrome virus (WSSV) challenges.  

Science.gov (United States)

The present study reports the influence of salinity (5, 15, 25 and 35?g/L) on the biochemical and immune parameters of Fenneropenaeus indicus challenged with 5. 5?×?10(4) copy number of White Spot Syndrome Virus. F. indicus that were reared in 25?g/L, were injected with WSSV, and then transferred to 5, 15, 25 (control) and 35g/L were examined for total haemocyte count (THC), phenoloxidase (PO) activity, superoxide dismutase (SOD) activity, alkaline phosphatase assay and acid phosphatase assay after 0?120?h. It was concluded that the F. indicus transferred from 25?g/L to low and higher salinity levels (5?g/L, 15?g/L and 35?g/L) had reduced immune indices and decreased resistance against WSSV infection. After 120?h, the mortality rate in WSSV injected F. indicus experimental groups (5?g/L and 35?g/L) was significantly higher than in F. indicus exposed to 25?g/L and 15?g/L salinities. During the experimental period (0-120?h), biochemical variables viz. total protein, total carbohydrate, and total lipid levels were measured in the hemolymph of both experimental and control groups. Acute salinity changes induced an increase in protein variations across the salinity ranges in shrimps. The THC, and PO activity, decreased significantly while SOD activity, alkaline phosphatase activity, and acid phosphatase activity was increased in the shrimp held at lower salinity of 5?g/L, 15?g/L and 35?g/L after 24?h. PMID:23586689

Vaseeharan, Baskaralingam; Ramasamy, Palaniappan; Wesley, Godwin; Chen, Jiann-Chu

2013-04-16

202

Visual detection of white spot syndrome virus using DNA-functionalized gold nanoparticles as probes combined with loop-mediated isothermal amplification.  

Science.gov (United States)

The integration of loop-mediated isothermal amplification (LAMP) and DNA-functionalized AuNPs as visual detection probes (LAMP-AuNPs) was developed and applied for the detection of white spot syndrome virus (WSSV) from Penaeid shrimp in this study. The principle of this combination assay relies on the basis of stability characteristics of the DNA-functionalized AuNPs upon hybridization with the complementary target DNA toward salt-induced aggregation. If the detected target DNA is not complementary to the ssDNA probes, the DNA-functionalized AuNPs will be aggregated due to the screening effect of salt, resulting in the change of solution color from red to blue/gray and shift of the surface plasmon peak to longer wavelength. While the DNA-functionalized AuNPs are perfectly matched to the detected target DNA, the color of solution still remains red in color and no surface plasmon spectral shift. This assay provides simply technique, time-saving and its detection results could be achieved qualitatively and quantitatively by visualization using the naked eye due to the colorimetric change and by measurement using the UV-vis spectroscopy due to the surface plasmon spectral shift, respectively. In this study, LAMP-AuNPs assay was successfully developed with the detection of WSSV-LAMP generated product at 0.03 ?g/reaction, and showed the sensitivity of 2 × 10(2) copies WSSV plasmid DNA, that is comparable to the most sensitive method reported to date. The LAMP-AuNPs assay described in this study revealed a highly sensitive, rapid and reliable diagnostic protocol for detection of WSSV. This technique has a potential as a routine method for assessing the infectious diseases in Penaeid shrimp not only for WSSV, but also for other shrimp pathogens, and can be useful tool in field conditions for the diagnosis or surveillance programs. PMID:23211683

Seetang-Nun, Yortyot; Jaroenram, Wansadaj; Sriurairatana, Siriporn; Suebsing, Rungkarn; Kiatpathomchai, Wansika

2012-12-01

203

Visual detection of white spot syndrome virus using DNA-functionalized gold nanoparticles as probes combined with loop-mediated isothermal amplification.  

UK PubMed Central (United Kingdom)

The integration of loop-mediated isothermal amplification (LAMP) and DNA-functionalized AuNPs as visual detection probes (LAMP-AuNPs) was developed and applied for the detection of white spot syndrome virus (WSSV) from Penaeid shrimp in this study. The principle of this combination assay relies on the basis of stability characteristics of the DNA-functionalized AuNPs upon hybridization with the complementary target DNA toward salt-induced aggregation. If the detected target DNA is not complementary to the ssDNA probes, the DNA-functionalized AuNPs will be aggregated due to the screening effect of salt, resulting in the change of solution color from red to blue/gray and shift of the surface plasmon peak to longer wavelength. While the DNA-functionalized AuNPs are perfectly matched to the detected target DNA, the color of solution still remains red in color and no surface plasmon spectral shift. This assay provides simply technique, time-saving and its detection results could be achieved qualitatively and quantitatively by visualization using the naked eye due to the colorimetric change and by measurement using the UV-vis spectroscopy due to the surface plasmon spectral shift, respectively. In this study, LAMP-AuNPs assay was successfully developed with the detection of WSSV-LAMP generated product at 0.03 ?g/reaction, and showed the sensitivity of 2 × 10(2) copies WSSV plasmid DNA, that is comparable to the most sensitive method reported to date. The LAMP-AuNPs assay described in this study revealed a highly sensitive, rapid and reliable diagnostic protocol for detection of WSSV. This technique has a potential as a routine method for assessing the infectious diseases in Penaeid shrimp not only for WSSV, but also for other shrimp pathogens, and can be useful tool in field conditions for the diagnosis or surveillance programs.

Seetang-Nun Y; Jaroenram W; Sriurairatana S; Suebsing R; Kiatpathomchai W

2013-04-01

204

Expression kinetics of ?-integrin in Chinese shrimp (Fenneropenaeus chinensis) hemocytes following infection with white spot syndrome virus.  

UK PubMed Central (United Kingdom)

Our previous study has demonstrated that an integrin ? subunit of Chinese shrimp (Fenneropenaeus chinensis) (Fc?Int) involved in WSSV infection. In order to further elucidate the potential role of the Fc?Int in the WSSV infection, expression response of Fc?Int to WSSV infection in shrimp hemocytes was investigated after intra-muscular injection with the virus. Following time-course hemocytes sampling, the expression variation of Fc?Int in hemocytes was examined by flow cytometric immunofluorescence assay (FCIFA) and enzyme linked immunosorbent assay (ELISA) using the monoclonal antibody (Mab) 2C5 against Fc?Int, which was successfully produced with recombinant partial Fc?Int and exhibited binding to a 120 kDa hemocyte protein. Meanwhile, the dynamic state of Fc?Int mRNA level and WSSV copies in hemocytes were determined by quantitative real-time PCR. The result of FCIFA showed that Fc?Int was mainly expressed on the semi-granular and granular cells, which was down-regulated at 6 h post infection (p.i.), and significantly increased to the peak level at 12 h p.i., then decreased to the control level by 24 h. However, Fc?Int on the hyaline cells was lowly expressed and didn't show active response to the viral infection. The variation of Fc?Int concentrations in total hemocytes determined by ELISA was roughly in accordance with the changing tendency of Fc?Int expressed on the semi-granular and granular cells. Fc?Int mRNA level in total hemocytes was significantly up-regulated to the peak level at 12 h p.i. Moreover, the number of WSSV copies in hemocytes began to exhibit a significant increase at 24 h p.i. The present study demonstrated that WSSV infection would induce a differential regulation of Fc?Int expression in different type hemocytes, which provided valuable evidences for the close correlation between Fc?Int and WSSV infection.

Zhong R; Tang X; Zhan W; Xing J; Sheng X

2013-08-01

205

Pre-exposure to infectious hypodermal and haematopoietic necrosis virus or to inactivated white spot syndrome virus (WSSV) confers protection against WSSV in Penaeus vannamei (Boone) post-larvae.  

UK PubMed Central (United Kingdom)

Larvae and post-larvae of Penaeus vannamei (Boone) were submitted to primary challenge with infectious hypodermal and haematopoietic necrosis virus (IHHNV) or formalin-inactivated white spot syndrome virus (WSSV). Survival rate and viral load were evaluated after secondary per os challenge with WSSV at post-larval stage 45 (PL45). Only shrimp treated with inactivated WSSV at PL35 or with IHHNV infection at nauplius 5, zoea 1 and PL22 were alive (4.7% and 4%, respectively) at 10 days post-infection (p.i.). Moreover, at 9 days p.i. there was 100% mortality in all remaining treatments, while there was 94% mortality in shrimp treated with inactivated WSSV at PL35 and 95% mortality in shrimp previously treated with IHHNV at N5, Z1 and PL22. Based on viral genome copy quantification by real-time PCR, surviving shrimp previously challenged with IHHNV at PL22 contained the lowest load of WSSV (0-1x10(3) copies microg-1 of DNA). In addition, surviving shrimp previously exposed to inactivated WSSV at PL35 also contained few WSSV (0-2x10(3) copies microg-1 of DNA). Consequently, pre-exposure to either IHHNV or inactivated WSSV resulted in slower WSSV replication and delayed mortality. This evidence suggests a protective role of IHHNV as an interfering virus, while protection obtained by inactivated WSSV might result from non-specific antiviral immune response.

Melena J; Bayot B; Betancourt I; Amano Y; Panchana F; Alday V; Calderón J; Stern S; Roch P; Bonami JR

2006-10-01

206

Pre-exposure to infectious hypodermal and haematopoietic necrosis virus or to inactivated white spot syndrome virus (WSSV) confers protection against WSSV in Penaeus vannamei (Boone) post-larvae.  

Science.gov (United States)

Larvae and post-larvae of Penaeus vannamei (Boone) were submitted to primary challenge with infectious hypodermal and haematopoietic necrosis virus (IHHNV) or formalin-inactivated white spot syndrome virus (WSSV). Survival rate and viral load were evaluated after secondary per os challenge with WSSV at post-larval stage 45 (PL45). Only shrimp treated with inactivated WSSV at PL35 or with IHHNV infection at nauplius 5, zoea 1 and PL22 were alive (4.7% and 4%, respectively) at 10 days post-infection (p.i.). Moreover, at 9 days p.i. there was 100% mortality in all remaining treatments, while there was 94% mortality in shrimp treated with inactivated WSSV at PL35 and 95% mortality in shrimp previously treated with IHHNV at N5, Z1 and PL22. Based on viral genome copy quantification by real-time PCR, surviving shrimp previously challenged with IHHNV at PL22 contained the lowest load of WSSV (0-1x10(3) copies microg-1 of DNA). In addition, surviving shrimp previously exposed to inactivated WSSV at PL35 also contained few WSSV (0-2x10(3) copies microg-1 of DNA). Consequently, pre-exposure to either IHHNV or inactivated WSSV resulted in slower WSSV replication and delayed mortality. This evidence suggests a protective role of IHHNV as an interfering virus, while protection obtained by inactivated WSSV might result from non-specific antiviral immune response. PMID:17026668

Melena, J; Bayot, B; Betancourt, I; Amano, Y; Panchana, F; Alday, V; Calderón, J; Stern, S; Roch, Ph; Bonami, J-R

2006-10-01

207

Histopathological study of white and black spot disease of shrimp Penaeus chinensis  

UK PubMed Central (United Kingdom)

Histopathology of white and black spot disease on shrimp Penaeus chinensis was studied with light and electron microscope examination .The results showed that there was no obvious or regular histopathological change in most part of tissues(hepatopancreas, gill, blood, stomach, gut, muscle, etc.)in diseased shrimp. Though about 37%of gills had granulations, and 41%of hepatopancreas were infected with HPV(Hepatpancreatic Virus), that hadno direct relation with white and black spot disease.White and black spot mainly appeared on both sides of each abdomen segment, tail fan, both sides of carapace ,antennal scale and rostrum etc, which resulted from deposit of brown and black melanin pigment. According to the analysis, the melanin was formed by phenoloxydase stimulating relative sbustrates. Both enzyme and substrates were considered from blood.Relationships between formation of white and black spot and physiological metabolism were discussed.

Cai Shengli; Wang Chongming; Yang Conghai

1999-01-01

208

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

Scientific Electronic Library Online (English)

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

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

2009-04-01

209

Neuroendocrine responses of a crustacean host to viral infection: effects of infection of white spot syndrome virus on the expression and release of crustacean hyperglycemic hormone in the crayfish Procambarus clarkii.  

Science.gov (United States)

The objectives of the present study were to characterize the changes in crustacean hyperglycemic hormone (CHH) transcript and peptide levels in response to infection of white spot syndrome virus (WSSV) in a crustacean, Procambarus clarkii. After viral challenge, significant increase in virus load began at 24 h post injection (hpi) and the increase was much more substantial at 48 and 72 hpi. The hemolymph CHH levels rapidly increased after viral challenge; the increase started as early as 3 hpi and lasted for at least 2 d after the challenge. In contrast, the hemolymph glucose levels did not significantly changed over a 2 d period in the WSSV-infected animals. The CHH transcript and peptide levels in tissues were also determined. The CHH transcript levels in the eyestalk ganglia (the major site of CHH synthesis) of the virus-infected animals did not significantly change over a 2 d period and those in 2 extra-eyestalk tissues (the thoracic ganglia and cerebral ganglia) significantly increased at 24 and 48 hpi. The CHH peptide levels in the eyestalk ganglia of the virus-infected animals significantly decreased at 24 and 48 hpi and those in the thoracic ganglia and cerebral ganglia remained unchanged over a 2 d period. These data demonstrated a WSSV-induced increase in the release of CHH into hemolymph that is rapid in onset and lasting in duration. Changes in the CHH transcript and peptide levels implied that the WSSV-induced increase in hemolymph CHH levels primarily resulted from an enhanced release from the eyestalk ganglia, but the contribution of the 2 extra-eyestalk tissues to hemolymph pool of CHH increased as viral infection progressed. The combined patterns of change in the hemolymph glucose and CHH levels further suggest that the virus-enhanced CHH release would lead to higher glycolytic activity and elevated glucose mobilization presumably favorable for viral replication. PMID:23174320

Lin, Ling-Jiun; Chen, Yan-Jhou; Chang, Yun-Shiang; Lee, Chi-Ying

2012-11-19

210

Neuroendocrine responses of a crustacean host to viral infection: effects of infection of white spot syndrome virus on the expression and release of crustacean hyperglycemic hormone in the crayfish Procambarus clarkii.  

UK PubMed Central (United Kingdom)

The objectives of the present study were to characterize the changes in crustacean hyperglycemic hormone (CHH) transcript and peptide levels in response to infection of white spot syndrome virus (WSSV) in a crustacean, Procambarus clarkii. After viral challenge, significant increase in virus load began at 24 h post injection (hpi) and the increase was much more substantial at 48 and 72 hpi. The hemolymph CHH levels rapidly increased after viral challenge; the increase started as early as 3 hpi and lasted for at least 2 d after the challenge. In contrast, the hemolymph glucose levels did not significantly changed over a 2 d period in the WSSV-infected animals. The CHH transcript and peptide levels in tissues were also determined. The CHH transcript levels in the eyestalk ganglia (the major site of CHH synthesis) of the virus-infected animals did not significantly change over a 2 d period and those in 2 extra-eyestalk tissues (the thoracic ganglia and cerebral ganglia) significantly increased at 24 and 48 hpi. The CHH peptide levels in the eyestalk ganglia of the virus-infected animals significantly decreased at 24 and 48 hpi and those in the thoracic ganglia and cerebral ganglia remained unchanged over a 2 d period. These data demonstrated a WSSV-induced increase in the release of CHH into hemolymph that is rapid in onset and lasting in duration. Changes in the CHH transcript and peptide levels implied that the WSSV-induced increase in hemolymph CHH levels primarily resulted from an enhanced release from the eyestalk ganglia, but the contribution of the 2 extra-eyestalk tissues to hemolymph pool of CHH increased as viral infection progressed. The combined patterns of change in the hemolymph glucose and CHH levels further suggest that the virus-enhanced CHH release would lead to higher glycolytic activity and elevated glucose mobilization presumably favorable for viral replication.

Lin LJ; Chen YJ; Chang YS; Lee CY

2013-02-01

211

Reducing white spot lesion incidence during fixed appliance therapy.  

UK PubMed Central (United Kingdom)

Fixed orthodontic appliances are commonly used in contemporary orthodontic treatment and can be associated with the development of white spot lesions on the teeth. These lesions can be detrimental to both the aesthetics and health of the teeth so prevention is better than cure and patient selection is critical. This paper discusses predictors of development in addition to methods to help prevent white spot lesions during fixed appliance therapy. Recommendations for oral hygiene regimes during fixed orthodontic appliance treatment are given, the development of white spot lesions (WSLs) described and ways to predict their occurrence identified. Clinical Relevance: Most general dental practitioners will have patients who are considering orthodontic treatment or are wearing fixed orthodontic appliances and so are at increased risk of developing WSLs. It is therefore important they are aware of predictors and ways to prevent WSLs.

Greene LE; Bearn DR

2013-07-01

212

Distribution of White Spots after Debanding in Orthodontic Patients  

Directory of Open Access Journals (Sweden)

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.

F. Arbabzadeh Zavareh; M. Bouzari; H. Nasr; MJ. Kharazifard

2006-01-01

213

Nucleotide sequence variations of the major structural proteins (VP15, VP19, VP26 and VP28) of white spot syndrome virus (WSSV), a pathogen of cultured Litopenaeus vannamei in Mexico.  

Science.gov (United States)

White spot syndrome virus (WSSV) was first reported in farmed Litopenaeus vannamei stocks in Sinaloa and Sonora, Mexico during 1999 and continues to cause severe shrimp losses. WSSV genes encoding nucleocapsid (VP26 and VP15) and envelope proteins (VP19 and VP28) of a Mexican isolate were cloned in the pMosBlue vector. The nucleotide sequences of these genes were compared with WSSV isolates in GenBank. VP15 is highly conserved, and VP26 showed 99% homology to a Chinese isolate. The VP28 fragment demonstrated 100% homology to the majority of the isolates analysed (UniProt accession no. Q91CB7), differing from two Indian WSSV and one Chinese WSSV isolates by two non-conserved and one conserved replacements, respectively. Because of their highly conserved nature, these three structural proteins are good candidates for the development of antibody-based WSSV diagnostic tools or for the production of recombinant protein vaccines to stimulate the quasi-immune response of shrimp. In contrast, VP19 of the Mexican isolate was distinguishable from almost all isolates tested, including an American strain of WSSV (US98/South Carolina, GenBank accession no. AAP14086). Although homology was found with isolates from Taiwan (GenBank accession no. AAL89341) and India (GenBank accession no. AAW67477), VP19 may have application as a genetic marker. PMID:18261033

Molina-Garza, Z J; Galaviz-Silva, L; Rosales-Encinas, J L; Alcocer-González, J M

2008-03-01

214

Nucleotide sequence variations of the major structural proteins (VP15, VP19, VP26 and VP28) of white spot syndrome virus (WSSV), a pathogen of cultured Litopenaeus vannamei in Mexico.  

UK PubMed Central (United Kingdom)

White spot syndrome virus (WSSV) was first reported in farmed Litopenaeus vannamei stocks in Sinaloa and Sonora, Mexico during 1999 and continues to cause severe shrimp losses. WSSV genes encoding nucleocapsid (VP26 and VP15) and envelope proteins (VP19 and VP28) of a Mexican isolate were cloned in the pMosBlue vector. The nucleotide sequences of these genes were compared with WSSV isolates in GenBank. VP15 is highly conserved, and VP26 showed 99% homology to a Chinese isolate. The VP28 fragment demonstrated 100% homology to the majority of the isolates analysed (UniProt accession no. Q91CB7), differing from two Indian WSSV and one Chinese WSSV isolates by two non-conserved and one conserved replacements, respectively. Because of their highly conserved nature, these three structural proteins are good candidates for the development of antibody-based WSSV diagnostic tools or for the production of recombinant protein vaccines to stimulate the quasi-immune response of shrimp. In contrast, VP19 of the Mexican isolate was distinguishable from almost all isolates tested, including an American strain of WSSV (US98/South Carolina, GenBank accession no. AAP14086). Although homology was found with isolates from Taiwan (GenBank accession no. AAL89341) and India (GenBank accession no. AAW67477), VP19 may have application as a genetic marker.

Molina-Garza ZJ; Galaviz-Silva L; Rosales-Encinas JL; Alcocer-González JM

2008-03-01

215

Mutations in MITF and PAX3 cause "splashed white" and other white spotting phenotypes in horses.  

UK PubMed Central (United Kingdom)

During fetal development neural-crest-derived melanoblasts migrate across the entire body surface and differentiate into melanocytes, the pigment-producing cells. Alterations in this precisely regulated process can lead to white spotting patterns. White spotting patterns in horses are a complex trait with a large phenotypic variance ranging from minimal white markings up to completely white horses. The "splashed white" pattern is primarily characterized by an extremely large blaze, often accompanied by extended white markings at the distal limbs and blue eyes. Some, but not all, splashed white horses are deaf. We analyzed a Quarter Horse family segregating for the splashed white coat color. Genome-wide linkage analysis in 31 horses gave a positive LOD score of 1.6 in a region on chromosome 6 containing the PAX3 gene. However, the linkage data were not in agreement with a monogenic inheritance of a single fully penetrant mutation. We sequenced the PAX3 gene and identified a missense mutation in some, but not all, splashed white Quarter Horses. Genome-wide association analysis indicated a potential second signal near MITF. We therefore sequenced the MITF gene and found a 10 bp insertion in the melanocyte-specific promoter. The MITF promoter variant was present in some splashed white Quarter Horses from the studied family, but also in splashed white horses from other horse breeds. Finally, we identified two additional non-synonymous mutations in the MITF gene in unrelated horses with white spotting phenotypes. Thus, several independent mutations in MITF and PAX3 together with known variants in the EDNRB and KIT genes explain a large proportion of horses with the more extreme white spotting phenotypes.

Hauswirth R; Haase B; Blatter M; Brooks SA; Burger D; Drögemüller C; Gerber V; Henke D; Janda J; Jude R; Magdesian KG; Matthews JM; Poncet PA; Svansson V; Tozaki T; Wilkinson-White L; Penedo MC; Rieder S; Leeb T

2012-01-01

216

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

Scientific Electronic Library Online (English)

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

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

2012-02-01

217

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

Directory of Open Access Journals (Sweden)

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

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

2012-01-01

218

Masking of white spot lesions by resin infiltration in vitro.  

UK PubMed Central (United Kingdom)

OBJECTIVES: The aim of this in vitro study was to evaluate the ability of one commercial and five experimental infiltrating resins (infiltrants) to camouflage enamel white spot lesions immediately after resin infiltration and after a staining period. METHODS: In each of 120 bovine enamel samples, two artificial caries lesions were created (windows A, C; pH=4.95, 50 days), whereas two windows were protected serving as sound controls (B, D). After etching windows C and D (37% phosphoric acid), specimens were randomly allocated to 6 groups. Either one of 5 experimental infiltrants or a commercial infiltrant (Icon, DMG) (refractive indices 1.50-1.55) was applied and light cured. After half of each specimen was polished, samples were remineralised (pH=7.0) and stained with tea and red wine for 50 days. Photographic images after various treatment steps were obtained. Colour differences (?E) of untreated (A) and treated lesions (C) as well as infiltrated sound enamel (D) were compared with untreated enamel (B). RESULTS: All infiltrants showed significantly better colour match with sound enamel (median ?E [25(th)/75(th) percentile]: 2.2 [1.5/3.1]) than untreated controls (9.3 [8.0/10.9]) (p<0.001, Wilcoxon, post-hoc Bonferroni). Moderate correlation between refractive index and ?E of infiltrated lesions was demonstrated (R(2)=0.43, p>0.05). Staining was significantly reduced for polished infiltrated lesions compared to untreated or infiltrated unpolished lesions (p<0.001). CONCLUSIONS: Resin infiltration is suitable to mask artificial white spot lesions. Polished infiltrated lesions are resistant to staining in vitro. CLINICAL SIGNIFICANCE: Resin infiltration is a micro-invasive approach to camouflage post-orthodontic white spot lesions.

Paris S; Schwendicke F; Keltsch J; Dörfer C; Meyer-Lueckel H

2013-04-01

219

Detection of red-spotted grouper nervous necrosis virus by loop-mediated isothermal amplification.  

UK PubMed Central (United Kingdom)

Red-spotted grouper nervous necrosis virus (RGNNV) causes high mortality in marine fish larvae cultured in China. To control better an outbreak of this virus, a rapid, specific and sensitive detection method based on loop-mediated isothermal amplification (LAMP) was developed. A set of four primers, two outer and two inner, was designed from RGNNV genome RNA. The LAMP reaction mix was optimized. The method was specific as no cross-reaction was observed between white spot syndrome virus, koi herpesvirus, infectious spleen and kidney necrosis virus, mud crab reovirus, and grass carp hemorrhage virus. The sensitivity of LAMP was 100-fold higher than the nested PCR in detecting the presence of RGNNV. RGNNV was detected in the brain of Trachinotus ovatus that showed typical symptoms of NNV infection, with the standardized LAMP procedure.

Xu HD; Feng J; Guo ZX; Ou YJ; Wang JY

2010-01-01

220

White spots on the mucosal surface of the duodenum in dogs with lymphocytic plasmacytic enteritis.  

Science.gov (United States)

Distended lacteals, described as expanded white villi in duodenum, are strongly indicative of primary intestinal lymphangiectasia. In the present study, we evaluated the significance of white spots present in the duodenal mucosa of dogs with lymphocytic plasmacytic enteritis (LPE). Fifty dogs with LPE were included in this study, and white spots were detected in the duodenal mucosa in 22 dogs during endoscopy. Hypoproteinemia was more frequent in dogs with white spots than in dogs without spots (p = 0.02). Serum protein and albumin concentration were significantly lower in LPE dogs with white spots (p = 0.038) compared to LPE dogs without white spots (p = 0.039). There was a significant correlation between white spots density and lymphatic dilatation histological scores (p = 0.023; ? = 0.481). These results suggest that the presence of white spots in the duodenal mucosa of dogs is not a finding exclusive for intestinal lymphangiectasia. Low serum protein and albumin concentrations together with lymphatic dilatation seem to be related to the presence of white spots in the duodenal mucosa of LPE dogs. PMID:21586876

García-Sancho, Mercedes; Sainz, Angel; Villaescusa, Alejandra; Rodríguez, Antonio; Rodríguez-Franco, Fernando

2011-06-01

 
 
 
 
221

White spots on the mucosal surface of the duodenum in dogs with lymphocytic plasmacytic enteritis.  

UK PubMed Central (United Kingdom)

Distended lacteals, described as expanded white villi in duodenum, are strongly indicative of primary intestinal lymphangiectasia. In the present study, we evaluated the significance of white spots present in the duodenal mucosa of dogs with lymphocytic plasmacytic enteritis (LPE). Fifty dogs with LPE were included in this study, and white spots were detected in the duodenal mucosa in 22 dogs during endoscopy. Hypoproteinemia was more frequent in dogs with white spots than in dogs without spots (p = 0.02). Serum protein and albumin concentration were significantly lower in LPE dogs with white spots (p = 0.038) compared to LPE dogs without white spots (p = 0.039). There was a significant correlation between white spots density and lymphatic dilatation histological scores (p = 0.023; ? = 0.481). These results suggest that the presence of white spots in the duodenal mucosa of dogs is not a finding exclusive for intestinal lymphangiectasia. Low serum protein and albumin concentrations together with lymphatic dilatation seem to be related to the presence of white spots in the duodenal mucosa of LPE dogs.

García-Sancho M; Sainz A; Villaescusa A; Rodríguez A; Rodríguez-Franco F

2011-06-01

222

White-spot lesions and gingivitis microbiotas in orthodontic patients.  

UK PubMed Central (United Kingdom)

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 < 0.01), and Bifidobacterium Cluster 1 (p = 0.11), and by qPCR, Streptococcus mutans (p = 0.008) and Scardovia wiggsiae (p = 0.04) Taxa associated with 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.

Tanner AC; Sonis AL; Lif Holgerson P; Starr JR; Nunez Y; Kressirer CA; Paster BJ; Johansson I

2012-09-01

223

Prevalence of white spot lesion formation during orthodontic treatment.  

UK PubMed Central (United Kingdom)

PURPOSE: To quantify the prevalence of white spot lesions (WSLs) on the anterior teeth and, secondarily, to evaluate risk factors and predictors. MATERIALS AND METHODS: Digital photographs and records of 885 randomly chosen patients were evaluated before and after treatment. Chart information included gender, age, as well as banding and debanding dates. Fluorosis and oral hygiene before and after treatment were also evaluated. Preexisting and posttreatment WSLs were recorded and compared for all 12 anterior teeth. Risk ratios (RR) and absolute risk (AR) were calculated to determine the likelihood and risk of WSL formation. RESULTS: Overall, 23.4% of the patients developed at least one WSL during their course of treatment. Maxillary anterior teeth were affected more than mandibular teeth. The maxillary laterals and canines and the mandibular canines were the most susceptible. There was no significant difference in WSLs between genders. Fluorosis, treatment time in excess of 36 months, poor pretreatment hygiene, hygiene changes during treatment, and preexisting WSLs were all significantly (P < .05) related to the development of WSLs. The highest risk of developing WSLs was associated with preexisting WSLs (RR = 3.40), followed by declines in oral hygiene during treatment (RR = 3.12) and poor pretreatment oral hygiene (RR = 2.83). CONCLUSIONS: Nearly 25% of the patients developed WSLs while in treatment, depending on fluorosis, treatment time, preexisting WSLs, and oral hygiene. Orthodontists need to be mindful of these risk factors when making treatment decisions.

Julien KC; Buschang PH; Campbell PM

2013-07-01

224

[Some biologic characteristics of two tomato spotted wilt virus isolates].  

Science.gov (United States)

Biological properties of two tomato spotted wilt virus (TSWV) isolates from the tomatoes, grown on plantations of the Crimean tobacco-plant experimental stations have been investigated. These isolates were slightly differentiated by thermal inactivation point, by terms of virus storage in juice and by dilution end point. As a result the growth of the induced resistance effective with respect to TMV was observed in tobacco-plants of Immunny-580 and N. sanderae varieties affected by TSWV. PMID:16396117

Shepelevych, V V

225

Transgenic tomato hybrids resistant to tomato spotted wilt virus infection.  

Digital Repository Infrastructure Vision for European Research (DRIVER)

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...

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

226

A SYSTEM TO EXPLORE TOMATO SPOTTED WILT VIRUS GENE FUNCTION  

Science.gov (United States)

Tomato spotted wilt virus (TSWV) has one negative sense (L) and two ambisense (M and S) RNAs. The lack of a tractable TSWV reverse genetics system has impeded direct demonstration of gene function and prevented the usual types of pathogen characterization. The TSWV NSm (non-structural) protein has...

227

Involvement of Beet western yellows virus, Cauliflower mosaic virus, and Turnip mosaic virus in Internal Disorders of Stored White Cabbage.  

Science.gov (United States)

ABSTRACT Experiments over two growing seasons clearly showed that Turnip mosaic virus (TuMV) infection was associated with internal necrosis (sunken necrotic spots 5 to 10 mm in diameter) and Beet western yellows virus (BWYV) infection was associated with collapse of leaf tissue at the margins (tipburn) in heads of stored white cabbage (Brassica oleracea var. capitata). Virtually no tipburn was seen in cv. Polinius, whereas cv. Impala was affected severely. Internal necrotic spots were seen in both cultivars. BWYV appeared to interact with TuMV. Plants infected with both viruses showed a lower incidence of external symptoms and had less internal necrosis than plants infected with TuMV alone. Cauliflower mosaic virus (CaMV) did not induce significant amounts of internal necrosis or tipburn, but did, in most cases, exacerbate symptoms caused by TuMV and BWYV. BWYV-induced tipburn worsened significantly during storage. Post-transplanting inoculation with TuMV induced more internal necrosis than pre-transplant inoculation. There was a significant association between detection of TuMV just prior to harvest and subsequent development of internal necrotic spots. Individually, all three viruses significantly reduced the yield of cv. Polinius, whereas only BWYV and CaMV treatments reduced the yield of cv. Impala. PMID:18942959

Hunter, P J; Jones, J E; Walsh, J A

2002-08-01

228

Transmission of cucumber leaf spot virus by Olpidium radicale.  

UK PubMed Central (United Kingdom)

The ability of zoospores of four cultures of Olpidium radicale and one of O. brassicae to transmit viruses acquired in vitro from dilute virus solutions was compared. Transmission was demonstrated by infectivity and serological assays of the roots of cucumber seedlings 6 days after inoculation. A bulk culture of O. radicale, from cucumber plant roots collected near Nantes, France, a single-sporangial culture derived from it, and a single-sporangial culture from melon plant roots collected near Woodland, California, U.S.A., transmitted cucumber leaf spot virus (CLSV) and the cucumber fruit streak strain of CLSV (CLSV-FS). A bulk culture of O. radicale from melon plant roots collected at Montfavet, France, did not transmit CLSV or CLSV-FS. All four cultures transmitted cucumber necrosis and melon necrotic spot viruses, used as positive controls, but they did not transmit cucumber soil-borne, squash necrosis, petunia asteroid mosaic or tobacco necrosis viruses. In each of the trials a single-sporangial culture of O. brassicae from lettuce plant grown in California transmitted only tobacco necrosis virus.

Campbell RN; Lecoq H; Wipf-Scheibel C; Sim ST

1991-12-01

229

Transmission of cucumber leaf spot virus by Olpidium radicale.  

Science.gov (United States)

The ability of zoospores of four cultures of Olpidium radicale and one of O. brassicae to transmit viruses acquired in vitro from dilute virus solutions was compared. Transmission was demonstrated by infectivity and serological assays of the roots of cucumber seedlings 6 days after inoculation. A bulk culture of O. radicale, from cucumber plant roots collected near Nantes, France, a single-sporangial culture derived from it, and a single-sporangial culture from melon plant roots collected near Woodland, California, U.S.A., transmitted cucumber leaf spot virus (CLSV) and the cucumber fruit streak strain of CLSV (CLSV-FS). A bulk culture of O. radicale from melon plant roots collected at Montfavet, France, did not transmit CLSV or CLSV-FS. All four cultures transmitted cucumber necrosis and melon necrotic spot viruses, used as positive controls, but they did not transmit cucumber soil-borne, squash necrosis, petunia asteroid mosaic or tobacco necrosis viruses. In each of the trials a single-sporangial culture of O. brassicae from lettuce plant grown in California transmitted only tobacco necrosis virus. PMID:1765774

Campbell, R N; Lecoq, H; Wipf-Scheibel, C; Sim, S T

1991-12-01

230

Detection of the L protein of tomato spotted wilt virus.  

UK PubMed Central (United Kingdom)

The 5'-terminal and 3'-terminal parts of the single open reading frame (ORF) in the L RNA of tomato spotted wilt virus (TSWV) were expressed using a prokaryotic expression system. Using antibodies raised against the translational products obtained a 330-kDa protein could be specifically detected in preparations of purified virions and in nucleocapsid preparations from TSWV-infected leaf tissue. The results obtained indicate that the L protein of TSWV, though much larger than that of the animal-infecting bunyaviruses, is present in virus particles in an unprocessed, intact form.

van Poelwijk F; Boye K; Oosterling R; Peters D; Goldbach R

1993-11-01

231

Frankliniella cephalica, a new vector for Tomato spotted wilt virus.  

UK PubMed Central (United Kingdom)

Frankliniella cephalica (Crawford) is an invasive species of thrips found in the islands of Yaeyama in the Okinawa Prefecture, Japan. During the late 1990s to early 2000s, a species of thrips was isolated from wild flowers of Bidens pilosa L. and Ipomoea batatas L. growing close to cultivated fields. They were subsequently identified as F. cephalica using fine morphological characteristics with the help of Steve Nakahara (U.S. Department of Agriculture, Beltsville, MD) and Laurence Mound (CSIRO, Australia). Voucher specimens were deposited in the Laboratory of Insect Resources, Faculty of Agriculture, Tokyo University of Agriculture by Shuji Okajima (2). We investigated the ability of F. cephalica to vector Tomato spotted wilt virus (TSWV) by experimentally determining virus transmission efficiency. Newly hatched larvae as much as 12 h old underwent a viral acquisition-access period (AAP) of 24 h, during which they fed on the leaves of Datura stramonium infected with TSWV-O, a Japanese type isolate. Transmission efficiency of adults 4 days after emergence from molt (14 days after the AAP) was determined by a petunia leaf disk assay (3) in which the adults were individually allowed to feed for successive 24-h inoculation access periods (IAP) on two different leaf disks of Petunia × hybrida cv. Polo Blue. Transmission of the virus by the adults was considered positive if at least one of the leaf disks showed viral necrotic spot. We tested 20 randomly selected leaf disks with clear necrotic spots using a simplified rapid immunofilter paper assay. All selected disks were positive for TWSV. The transmission efficiencies were 24.6% for female (n = 57) and 54.4% for male (n = 125) adults. The efficiency was significantly different between sexes (Fisher's exact probability test, P < 0.001). We also examined changes in the virus infection site at different developmental stages in thrips using immunofluorescence microscopy with a polyclonal antibody to N protein of the virus (4). After a 6-h AAP feeding by first instar larvae, the virus was found initially to infect the epithelial cells and then spread throughout the midgut tissue in the second instar larvae 5 days after acquisition of the virus. In viruliferous adults, the virus was present in the salivary glands and on the basement membrane of the midgut tissue. These data indicate that F. cephalica is a new insect vector for TSWV. F. cephalica is a major insect pest of tropical crops in tropical and subtropical coastal belts (1). The presence of a thrips vector in weed hosts surrounding cultivated fields might increase the chance of crops in this habitat becoming infected with viruses.

Ohnishi J; Katsuzaki H; Tsuda S; Sakurai T; Akutsu K; Murai T

2006-05-01

232

Novel treatment of white spot lesions: A report of two cases.  

UK PubMed Central (United Kingdom)

This case report describes a technique used to treat smooth surface white spot lesions microinvasively. It is based on the infiltration of an initial enamel caries lesion with low-viscosity light-curing resins called infiltrants. The surface layer is eroded and desiccated, followed by resin infiltrant application. The resin penetrates into the lesion microporosities driven by capillary force and is hardened by light curing. Infiltrated lesions lose their whitish appearance and look similar to sound enamel. Additionally, the treatment prevents lesion progression. This technique might be an alternative to microabrasion and restorative treatment in treating of white spot lesions of esthetically relevant teeth.

Shivanna V; Shivakumar B

2011-10-01

233

Novel treatment of white spot lesions: A report of two cases.  

Science.gov (United States)

This case report describes a technique used to treat smooth surface white spot lesions microinvasively. It is based on the infiltration of an initial enamel caries lesion with low-viscosity light-curing resins called infiltrants. The surface layer is eroded and desiccated, followed by resin infiltrant application. The resin penetrates into the lesion microporosities driven by capillary force and is hardened by light curing. Infiltrated lesions lose their whitish appearance and look similar to sound enamel. Additionally, the treatment prevents lesion progression. This technique might be an alternative to microabrasion and restorative treatment in treating of white spot lesions of esthetically relevant teeth. PMID:22144817

Shivanna, Vasundhara; Shivakumar, B

2011-10-01

234

In vitro interactions between the surfaces of enamel white spots and calcifying solutions.  

UK PubMed Central (United Kingdom)

The treatment of white spot enamel lesions with a saturated DCPD solution followed by a high fluoride topical application appeared to produce a surface coating consisting mainly of spherical particles that filled in the honeycombed-etched surface of the untreated enamel white spot. The conditions used in these pilot studies were not ideal to take advantage of the DCPD and fluoride reaction to form FAP. Additional study is warranted on the solution composition and application time along with identification of the reaction products formed.

Wei SH; Wefel JS

1976-01-01

235

Pittosporum tobira: a new host for Tomato spotted wilt virus.  

UK PubMed Central (United Kingdom)

In July 1998, Pittosporum tobira shrubs, grown in a nursery in the Sharon Valley of Israel, developed foliar ring spots, mild mosaic, and tip necrosis. Of 15 samples tested for the presence of Tomato spotted wilt virus (TSWV) with a commercially available enzyme-linked immunosorbent assay (ELISA) kit (Loewe Biochemica, Otterfing, Germany), 14 were positive for TSWV. Virus in crude sap extracted from symptomatic tissue was mechanically transmitted to Emilia spp., Petunia hybrida, Nicotiana glutinosa, N. benthamiana, and N. rustica plants, which developed symptoms characteristic of TSWV infection (1). ELISA tests of leaf sap extracted from naturally infected P. tobira and mechanically inoculated indicator plants gave a strong positive reaction to TSWV. Leaf samples of P. tobira were analyzed by transmission electron microscopy in leaf-dip preparations and thin sections of leaf tissues. Virus particles typical of a tospovirus were observed only in samples taken from symptomatic leaves. Primers specific to the nucleocapsid gene of TSWV were used in a reverse transcription-polymerase chain reaction (RT-PCR) assay to verify the presence of TSWV. RT-PCR gave an expected PCR product of ?850 bp. The amplicon was cloned in the pGEM-T vector, and the recombinant clone was sequenced. The sequence of the cloned PCR product confirmed the identity of TSWV, verifying TSWV infection of P. tobira. This is the first report of infection of P. tobira by TSWV.

Gera A; Kritzman A; Cohen J

2000-04-01

236

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

UK PubMed Central (United Kingdom)

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

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

2011-01-01

237

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

Science.gov (United States)

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

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

2011-01-01

238

Evaluation of 3 commercially available materials for resolution of white spot lesions.  

UK PubMed Central (United Kingdom)

INTRODUCTION: White spot lesions are often seen on the teeth after orthodontic treatment, resulting in unpleasant esthetics. The aim of this in-vitro study was to compare subjectively and objectively the esthetic outcomes of white spot lesions treated with 3 commercially available products that have been reported to have a positive effect on the remineralization of enamel. METHODS: Forty extracted premolars were randomly allocated into 1 of 4 groups (n = 10). The teeth were exposed to a demineralization solution at 37°C for 14 days to produce white spot lesions that were about 100 ?m deep. Each group was then randomly assigned to receive either control treatment with artificial saliva or treatment with 1 of the 3 commercially available products: Restore toothpaste (Dr. Collins Inc, Orange County, Calif), which contains NovaMin (Dr. Collins Inc); Prevident 5000 (Colgate, New York, NY); and MI Paste Plus (GC America, Alsip, Ill). All groups were evaluated 5 times at 6 time points during the study. RESULTS: The subjective and objective results were mixed within groups and between groups for the products tested. CONCLUSIONS: We found no conclusive evidence that any of these 3 materials produced more favorable esthetic white spot lesion remineralization results.

Ballard RW; Hagan JL; Phaup AN; Sarkar N; Townsend JA; Armbruster PC

2013-04-01

239

Virus diseases and stand decline in a white clover pasture.  

UK PubMed Central (United Kingdom)

Linear regression analysis was used to characterize the incidence of virus diseases in white clover, Trifolium repens, and associated changes in the white clover population in a grazed pasture through four growing seasons. The incidence of diseases caused by clover yellow vein virus, peanut stunt virus (PSV), and white clover mosaic virus increased linearly through the life of the white clover stand, coincident with a steady decline in the white clover population. The predominant virus detected throughout the study was PSV, which occurred in 80% of the white clover plants sampled in the spring of the fourth growing season. By that time, the clover population had declined significantly, and by the following fall, the clover had virtually disappeared from the pasture. This association provided evidence that PSV may be a major factor in the lack of persistence of white clover. A potentially important source of resistance to PSV was identified in a white clover clone exhibiting hypersensitivity to mechanical inoculation of PSV. Alfalfa mosaic virus and red clover vein mosaic virus were present in infected white clover but at relatively low incidence. Luteoviruses not previously reported from white clover in the Southeast also were detected.

McLaughlin MR; Pederson GA; Evans RR; Ivy RL

1992-02-01

240

New natural hosts of Tomato spotted wilt virus.  

UK PubMed Central (United Kingdom)

Tomato spotted wilt virus (TSWV) has caused epidemics in recent years in many crops throughout the Mediterranean Region. Tomato, pepper, and lettuce are the crops most affected in Spain. To determine the reservoir hosts for the virus in the area, 210 samples from 95 species of plants were collected and tested for TSWV by double-antibody sandwich enzyme-linked immunosorbent assay with commercial antisera (Loewe Biochemica, Germany: BR-01, serogroup I or TSWV-L). Twenty-one species tested positive, and among them were thirteen newly identified hosts for TSWV (1). Weed species were among the 13 new hosts and included Diplotaxis erucoides (L.) DC., Beta maritima L., Phragmites communis Trin., Malva sylvestris L., Sonchus arvensis L., Sorghum halepense L., Panicum repens L., Atriplex patula L., Coronopus squamatus (Forssk.) Ascherson, Cuscuta sp., Xanthium spinosum L., Suaeda vera J.F., and Ecballium elaterium (L.) A. Rich. Most of these plants were asymptomatic hosts, but the Sonchus sp. showed typical symptoms of TSWV, such as yellows, bronzing, ring spots, necrosis, curling of young leaves, and growth reduction. D. erucoides, B. maritima, M. sylvestris, X. spinosum, and E. elaterium showed chlorosis and growth reduction.

Jorda C; Font I; Lazaro A; Juarez M; Ortega A; Lacasa A

2000-04-01

 
 
 
 
241

Sexual selection for white tail spots in the barn swallow in relation to habitat choice by feather lice.  

UK PubMed Central (United Kingdom)

Many bird species have white spots in their tails or wing feathers, and such characters have been hypothesized to be either reliable signals (handicaps) or amplifiers that facilitate the message of a signal. In barn swallows, Hirundo rustica, the size of the white spots in the tail feathers is sexually dimorphic and positively correlated with feather length. We tested whether such spots act as handicaps or amplifiers. These white spots affect sexual selection in barn swallows, as shown by an experiment in which we randomly subjected males to (1) a considerable reduction of the size of all the spots by the use of a black permanent marker pen, (2) a small reduction of the size of the spots, or (3) no reduction. There was a positive association between spot size and the number of offspring produced per season. The white tail spots were preferred by feather-eating Mallophaga as a feeding site: holes made by Mallophaga were more abundant in the white spots than expected by chance. A habitat choice experiment with Mallophaga on barn swallow tail feathers revealed that they preferred white spots over black parts of the tail feathers. We therefore expected long-tailed male barn swallows to have more Mallophaga than short-tailed males. However, the opposite relationship was observed, indicating that long-tailed males may reliably signal their quality by the presence of large white tail spots without parasite damage. Thus white tail spots in barn swallows appear to be a reliable signal of phenotypic quality. Copyright 1999 The Association for the Study of Animal Behaviour.

Kose M; Mänd R; Møller AP

1999-12-01

242

Tomato Spotted Wilt Virus – One of the Most Destructive Plant Viruses  

Directory of Open Access Journals (Sweden)

Full Text Available Tomato spotted wilt virus (TSWV) has one of the largest host ranges among plant viruses and is widespread in all climates. TSWV is responsible for numerous epidemics in many parts of the world in different crops, mainly vegetables, tobacco and ornamentals. Its highly polyphagous nature, effectiveness of virus transmission by the thrips as its vectors, rapidity with which new variants arise, as well as difficulties in controlling the vectors make TSWV one of the most dangerous plant viruses. The ability of this virus to cause such severe losses on a broad range of crops, as well as its intriguing biological and molecular characteristics place TSWV amongst the most extensively studied plant viruses in the world at present. This paper provides a general overview of TSWV, encompassing all the major aspects of its biology and current knowledge on host range, symptomatology, molecular biology, vectorrelationship, control and diagnosis.

Branka Krsti?; Aleksandra Bulaji?; Ivana ?eki?; Janoš Berenji

2008-01-01

243

Variability in field response of peanut genotypes from the U.S. and China to tomato spotted wilt virus and leaf spots  

Science.gov (United States)

Tomato spotted wilt, caused by tomato spotted wilt virus (TSWV) and transmitted by thrips, and early leaf spot (Cercospora arachidicola) and late leaf spot (Cercosporidium personatum) are among the most important diseases of peanut (Arachis hypogaea) in the southeastern United States. The objective ...

244

First report of Cucumber leaf spot virus in Spain.  

UK PubMed Central (United Kingdom)

Cucumber leaf spot virus (CLSV), reclassified as a species in the new genus Aureusvirus (family Tombusviridae) (1), has ?30-nm isometric particles with a ?4.4-kb positive-sense, single-stranded RNA. CLSV is transmitted by the chytrid fungus Olpidium bornovanus. The virus has been reported in Germany, Great Britain, Greece, Jordan, and Saudi Arabia. During the fall of 2000, abundant chlorotic spots with necrotic centers were observed on the leaves of cucumber plants grown in a commercial greenhouse in Granada (southeastern Spain). When sap from collected leaves was used to mechanically inoculate cucumber, symptoms were reproduced and were suggestive of CLSV. Based on the nucleic acid sequence of CSLV (2), the following specific primers were designed: CLSVU1440 (5'-AAGGTAGGGGAGATCTTG-3') and CLSVA2160 (5'-GCTTCGGCTGATTCTGA-3'). When used in reverse transcription-polymerase chain reactions (RT-PCR), leaves expressing symptoms yielded amplification products of the expected size (720 bp). These products were cloned into a pGEM-T vector and sequenced (GenBank Accession No. AY038365). The similarity of the nucleic acid and derived amino acid sequences with the one published for CLSV (2) was 94.5 and 99.1%, respectively. The amino acid sequence was 86% identical to that of Pothos latent virus (GenBank Accession No. AJ243370). Ten cucumber plants grown in vermiculite supplemented with rhizosphere soil (1/30, vol/wt) from infected plants developed symptoms on leaves after 1 month and were positive for CLSV when leaf and root tissues were analyzed by RT-PCR and Southern blot hybridization. Plants grown in vermiculite alone did not become infected with CLSV. Microscopic examination of root tissue revealed O. bornovanus only in infected plants. To our knowledge, this is the first record of CSLV in Spain.

Segundo E; Janssen D; Velasco L; Ruiz L; Cuadrado IM

2001-10-01

245

Field evaluation for the combination of white mould and tomato spotted wilt disease resistance among peanut genotypes  

UK PubMed Central (United Kingdom)

White mould, stem rot, and southern blight are caused by the same soilborne fungus, Sclerotium rolfsii Sacc. and when combined with tomato spotted wilt caused by Tomato spotted wilt virus (TSWV), result in major disease problems in the cultivated peanut (Arachis hypogaea L.). These disease problems are universal and occur not only in the southeast U.S.A. but also in most peanut production areas. Consequently, there is an increasing need for genotypic resistance as the most cost effective way to reduce the financial loss due to the combination of these two diseases. During five years (2004-08), several advanced breeding lines and recently released peanut cultivars from the Georgia peanut breeding program were evaluated for combined resistance to both pathogens. The best combination of white mould and TSWV resistance and highest yield over multiple years was consistently found in the recently released medium-maturity, runner-type peanut cultivars, Georgia-07W, Georgia-03L, and AP-3. However, all of the other Georgia cultivars and advanced Georgia breeding lines were also found to be either similar or significantly better than the check cultivars C-99R and Florida-07.

Branch WD; Brenneman TB

2009-07-01

246

Effectiveness of tomato-spotted wilt virus management tactics.  

UK PubMed Central (United Kingdom)

Tomato-spotted wilt (TSW) is caused by the tomato-spotted wilt virus (TSWV) and is a major disease affecting the production of tomato and pepper in the Southeastern United States. Before initiating a multistate, regional project addressing this issue, a survey was conducted in North Carolina, South Carolina, Georgia, and Florida to assess the need for improved management of TSWV in these crops. We investigated farmer's stated effectiveness of four common TSWV management tactics (reflective mulch, resistant cultivar, imidacloprid, and Actiguard) in pepper and tomato production using logistic regression. We found that the odds that farmers were satisfied with the use of reflective mulch alone in controlling TSWV is 9-1, about one and a half times the amount obtained from using Actiguard alone. Moreover, the odds that farmers were satisfied with a practice that combines reflective mulch and Actiguard was far greater than that obtained from using each of the technique separately. We found some similarities between farmer's stated effectiveness and revealed effectiveness from experiments.

Awondo SN; Fonsah EG; Riley D; Abney M

2012-06-01

247

A compound Chinese herbal medicament preparation for preventing and treating white spot syndrome  

UK PubMed Central (United Kingdom)

A compound Chinese herbal medicament preparation for preventing and treating white spot syndrome is prepared by following raw materials by weight: 20-35 parts of Corydalis bungeana Turcz., 25-40 partsof radix isatidis, 15-40 parts of radix astragali, 20-40 parts of dyers woad leaf, 25-35 parts of wrinkled giant hyssop, 10-30 parts of Japanese eupatorium, 15-25 parts of roughhaired holly root and5-15 parts of Gynostemma pentaphylla. The compound Chinese herbal medicament preparation can effectively prevent and treat the white spot syndrome. The invention also discloses a forage comprising thecompound Chinese herbal medicament preparation and a method for feeding the same. The forage comprising the compound Chinese herbal medicament has simple processing technology and convenient feeding.The method for feeding the forage comprising the compound Chinese herbal medicament is scientific and reasonable and has a good antiviral effect.

ZHUOJIA LI; ZHIXUN GUO; HEIZHE LIN; QINGHUA YANG; HAIDONG XU

248

First Report of Melon necrotic spot virus in Panama  

UK PubMed Central (United Kingdom)

Melon (Cucumis melo L.) represents an important crop in Panama where 1,449 ha were cultivated in 2005 with 920.4 ha of this crop planted in Los Santos Province (southeast region of Panama). During April 2005 and January 2006, several melon plants in commercial fields in that area showed stem necrosis at the crown level, and less frequently, small necrotic spots on leaves. In some cases, wilting and plant death were observed. Symptoms were similar to those caused by the carmovirus Melon necrotic spot virus (MNSV). Cysts of Olpidium bornovanus also were observed in the roots of all affected melon plants. Roots from eight symptomatic plants collected in seven fields were positive using doubleantibody sandwich enzyme-linked immunosorbent assay (DAS-ELISA) with an antiserum specific for MNSV (BIO-RAD, Life Sciences, Barcelona, Spain). To confirm these results, total RNA was extracted from symptomatic plants and used in one-step reverse transcription-polymerase chain reaction (RT-PCR) with Platinum Taq (Invitrogen Life Technologies, Barcelona, Spain). MNSV specific primers designed to amplify a region of the coat protein gene were used in the assays. Amplicons of the expected size (651 bp) were generated from symptomatic plant tissue, but were not produced from healthy plants or the water used as negative controls. To establish the authenticity of this virus, RT-PCR products were purified with the High Pure PCR Product Purification Kit (Roche Diagnostics, Mannheim, Germany) and directly sequenced. Nucleotide sequences were analyzed by using the basic local alignment search tool (BLAST) (1). The primers produced two amplicons with different but similar sequences. One sequence (GenBank Accession No. DQ443546) showed 92% identity to the coat protein gene of the MNSV Spanish isolate (GenBank Accession No. AY330700) and the MNSV Dutch isolate (GenBank Accession No. M29671) and 88% identity to the Japanese isolate (GenBank Accession No. AB189944). The second sequence (GenBank Accession No. DQ443547) was 93% identical with the Spanish and Dutch MNSV isolates, 88% identical with the Japanese isolate, and 100% identical with sequences from commercial melon seed previously isolated in our laboratory (GenBank Accession No. DQ443545). Infected seed may be a concern with regard to long distance spread of the virus independent of the vector (3) and should be considered in disease management strategies. MNSV has been previously reported in Japan, the Netherlands, the United Kingdom, the United States (2), Guatemala (4), Mexico, Honduras, and Uruguay (C. Jordá, unpublished). To our knowledge, this is the first report of MNSV in Panama.

Herrera JA; Cebrian MC; Jorda C

2006-09-01

249

Cutaneous nerves in cafe au lait spots with white halos in infants with neurofibromatosis. An electron microscopic study.  

UK PubMed Central (United Kingdom)

BACKGROUND AND DESIGN: Although two cardinal skin manifestations of neurofibromatosis are cutaneous neurofibromas and cafe au lait spots, the pathogenesis of cafe au lait spots are very poorly known compared with that of cutaneous neurofibromas. Thus, the cafe au lait spots in two Japanese infants were clinically, histologically, and electron-microscopically investigated. OBSERVATIONS: Some of the cafe au lait spots in the mongolian spots were surrounded by white halos. Histologically, in the cafe au lait spots, the epidermal basal cells had abundant melanin pigment, but macromelanosomes were not seen throughout the epidermis. In the white halo, the epidermal basal cells had a small amount of melanin pigment. Electron microscopically, the cafe au lait spots and their white halos had many subepidermal and intraepidermal nerves that belonged to free nerve endings. All the cutaneous nerves were mature. Some of the intraepidermal nerves had partially or completely naked axons that contacted tightly with the cytomembranes of the basal keratinocytes. Some of the axons in the subepidermal nerves showed degenerative changes only in the white halos. No ultrastructural pathologic changes were observed in the melanocytes, the epidermal keratinocytes, or melanosomes in those cells in the cafe au lait spots and their white halos; also, dermal melanocytes were absent in the both areas. CONCLUSIONS: The increase of the cutaneous nerves and the absence of dermal melanocytes in the cafe au lait spots and their white halos may be considered as characteristic histologic cutaneous findings in infants with neurofibromatosis. However, no evidence indicates that the cutaneous nerves may participate closely in the pathogenesis of the white halos.

Mihara M; Nakayama H; Aki T; Inoue T; Shimao S

1992-07-01

250

Phylogenetic and recombination analysis of tomato spotted wilt virus.  

UK PubMed Central (United Kingdom)

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

Lian S; Lee JS; Cho WK; Yu J; Kim MK; Choi HS; Kim KH

2013-01-01

251

Phylogenetic and recombination analysis of tomato spotted wilt virus.  

Science.gov (United States)

Tomato spotted wilt virus (TSWV) severely damages and reduces the yield of many economically important plants worldwide. In this study, we determined the whole-genome sequences of 10 TSWV isolates recently identified from various regions and hosts in Korea. Phylogenetic analysis of these 10 isolates as well as the three previously sequenced isolates indicated that the 13 Korean TSWV isolates could be divided into two groups reflecting either two different origins or divergences of Korean TSWV isolates. In addition, the complete nucleotide sequences for the 13 Korean TSWV isolates along with previously sequenced TSWV RNA segments from Korea and other countries were subjected to phylogenetic and recombination analysis. The phylogenetic analysis indicated that both the RNA L and RNA M segments of most Korean isolates might have originated in Western Europe and North America but that the RNA S segments for all Korean isolates might have originated in China and Japan. Recombination analysis identified a total of 12 recombination events among all isolates and segments and five recombination events among the 13 Korea isolates; among the five recombinants from Korea, three contained the whole RNA L segment, suggesting reassortment rather than recombination. Our analyses provide evidence that both recombination and reassortment have contributed to the molecular diversity of TSWV. PMID:23696821

Lian, Sen; Lee, Jong-Seung; Cho, Won Kyong; Yu, Jisuk; Kim, Mi-Kyeong; Choi, Hong-Soo; Kim, Kook-Hyung

2013-05-17

252

A white spot disease-like syndrome in the Pacific blue shrimp (Litopenaeus stylirostris) as a form of bacterial shell disease  

Digital Repository Infrastructure Vision for European Research (DRIVER)

In May 1997, some white lesions evoking the white spot syndrome disease were observed in Litopenaeus stylirostris broodstock in New Caledonia. The occurrence of these lesions was neither associated with mortality, nor with histological evidence of white spot syndrome baculovirus (WSBV), The evidence...

Goarant Cyrille; Brizard Raphael; Marteau Anne-laure

253

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

Scientific Electronic Library Online (English)

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

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

2010-06-01

254

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

Directory of Open Access Journals (Sweden)

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

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

2010-01-01

255

Differential acquisition and transmission of Florida Tomato spotted wilt virus isolates by Western flower thrips  

Science.gov (United States)

Thrips-vectored Tomato spotted wilt virus (TSWV) is one of the most important insect-vectored plant pathogens globally. The virus host range encompasses many key vegetable, ornamental and agronomic crops. TSWV populations are highly heterogeneous, which has important implications for vector relati...

256

Tomato spotted wilt virus NSm protein domains involved in tubule formation,movement and symptoms  

Science.gov (United States)

Direct demonstration of Tomato spotted wilt virus (TSWV) gene function has been slowed by the absence of a reliable reverse genetics system. A Tobacco mosaic virus (TMV)-based expression system was previously used by us to demonstrate that the TSWV NSm protein is able to support cell-to-cell movemen...

257

Genetic characterization of Okinawan black rats showing coat color polymorphisms of white spotting and melanism.  

UK PubMed Central (United Kingdom)

We examined pelage color variation in wild populations of black rats (the Rattus rattus species complex) in the Yambaru forest area, northern Okinawa Island, Ryukyu Archipelago, Japan. Our field study revealed that 8.7% (38/438) and 0.2% (4/2500) of rats exhibited two types of coat color: white spotting and melanism, respectively. Using 34 representative animals, the phylogeography of the population was inferred using a nuclear gene marker, i.e., sequences (954 bp) of the melanocortin-1 receptor (Mc1r) gene responsible for the melanistic form in black rats. Four sequences from Okinawa were characterized as R. tanezumi, the Asian strain of black rat. Notably, neither of the phenotypic characters of white spotting or melanism was associated with the Mc1r haplotypes. Analysis of mitochondrial cytochrome b (Cytb) sequences (1140 bp) revealed that four haplotypes recovered from Okinawa clustered with the clade of R. tanezumi and differed by one or more bases from haplotypes at other localities in Japan and Asian countries. Thus, both variants may have arisen in the native rat population of Okinawa without interaction with the lineage of R. rattus, which exhibits a worldwide distribution and displays such coat color variants. The Yambaru population of black rats has thus experienced its own evolutionary history in allopatry for a substantial period of time (e.g., 10,000 years), which has preserved valuable genetic polymorphisms and will be useful for assessing the ecological consequences of genetic variation in natural populations.

Kambe Y; Nakata K; Yasuda SP; Suzuki H

2012-01-01

258

Sequence similarities between Raspberry leaf mottle virus, Raspberry leaf spot virus and the closterovirus Raspberry mottle virus  

UK PubMed Central (United Kingdom)

A sequencing study was performed to determine the relationship between Raspberry mottle virus (RMoV), a newly identified tentative closterovirus found in the United States, and Raspberry leaf mottle virus (RLMV) and Raspberry leaf spot virus (RLSV), which have been known for many years to be components of Raspberry mosaic disease (RMD) in the UK and Europe but which have not been characterised at the molecular level. Cloning and sequencing of cDNAs amplified by reverse transcription-PCR revealed the presence of closteroviruses with high sequence similarity to RMoV in infected plants from the SCRI Rubus virus collection, as well as in a number of samples collected from RMD-symptomatic raspberry plants located at different farms in Scotland and England. These results suggest that RMoV, RLMV and RLSV are isolates of the same virus and we propose that they all should be referred to as RLMV, which was the first of these viruses to be described. Many of the field samples were also infected with a second closterovirus isolate, parts of which could be amplified using RLMV-derived primers. The coat protein amino acid sequences of RLMV and the second virus (PM1) were only 78% identical, even though helicase domain and RNA-dependent RNA polymerase (RDRP) domain sequences were more than 97% identical between RLMV and PM1.

McGavin WJ; MacFarlane SA

2010-05-01

259

Detection of cymbidium mosaic virus, Odontoglossum ringspot virus, tomato spotted virus, and potyviruses infecting orchids in Hawaii.  

UK PubMed Central (United Kingdom)

Approximately 3,600 orchid plants representing 44 genera from three orchid collections, 22 commercial farms, and six nurseries on the islands of Oahu and Hawaii were tested for Cymbidium mosaic virus (CyMV), Odontoglossum ringspot virus (ORSV), tomato spotted wilt virus (TSWV), and potyviruses with enzyme-linked immunosorbent assay (ELISA). CyMV was detected in 59 samples and ORSV was detected in 23 samples, which represented 61 and 25% of the 44 genera surveyed, respectively. Double infection with both viruses occurred in 20 samples, representing 20% of the genera. CyMV and ORSV were detected in 29 and seven of the 31 sites surveyed, respectively. When 330 cloned orchid samples were tested, CyMV was detected in 45% and ORSV in 9% of the clones. Most commercial Dendrobium orchids grown in Hawaii are seed-propagated hybrids produced by the Horticulture Department at the University of Hawaii (UH). Only 4% of the 758 UH Dendrobium hybrids less than three years old were infected by CyMV. Of 2,381 UH Dendrobium hybrids more than 3 yr old, CyMV was detected in 94% of the samples from some farms, but only 2% from other farms. ORSV was not identified from any commercially grown UH Dendrobium hybrids. TSWV-infected Oncidium orchids, found in only one nursery in Hawaii, exhibited symptoms ranging from chlorotic ring spots to necrotic lesions 1–2 cm in diameter. The TSWV infection was localized in the ring spots and lesions. Dendrobium mosaic potyvirus was detected in Dendrobium superbum.

Hu JS; Ferreira S; Wang M; Xu MQ

1993-05-01

260

Identification of tomato spotted wilt-like virus on watermelon in Taiwan.  

UK PubMed Central (United Kingdom)

The causal agent of an unusual disease affecting watermelon (Citrullus lanatus) in Taiwan was identified as tomato spotted wilt-like virus in the tospovirus group on the basis of particle morphology, host reactions, serology, and transmission by thrips. Host symptoms included leaf crinkling, mottling, yellow spotting, short internodes, upright growth of younger branches, and tip necrosis. The systemic infection of cucurbits and transmissibility by the vector Thrips palmi distinguished this isolate from most other tomato spotted wilt virus (TSWV) isolates. Serological tests by enzyme-linked immunosorbent assay and western blotting with polyclonal and monoclonal antibodies demonstrated that the watermelon virus is serologically related to the TSWV that causes silver mottle on watermelon in Okinawa (Japan) and not to the tomato, lettuce, and amaryllis isolates of TSWV from other countries.

Yeh SD; Lin YC; Cheng YH; Jih CL; Chen MJ; Chen CC

1992-08-01

 
 
 
 
261

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

DEFF Research Database (Denmark)

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

Bröchner, Ann; Christensen, Carsten

2010-01-01

262

Novel variants in the KIT and PAX3 genes in horses with white-spotted coat colour phenotypes.  

UK PubMed Central (United Kingdom)

Variants in the EDNRB, KIT, MITF, PAX3 and TRPM1 genes are known to cause white spotting phenotypes in horses, which can range from the common white markings up to completely white horses. In this study, we investigated these candidate genes in 169 horses with white spotting phenotypes not explained by the previously described variants. We identified a novel missense variant, PAX3:p.Pro32Arg, in Appaloosa horses with a splashed white phenotype in addition to their leopard complex spotting patterns. We also found three novel variants in the KIT gene. The splice site variant c.1346+1G>A occurred in a Swiss Warmblood horse with a pronounced depigmentation phenotype. The missense variant p.Tyr441Cys was present in several part-bred Arabians with sabino-like depigmentation phenotypes. Finally, we provide evidence suggesting that the common and widely distributed KIT:p.Arg682His variant has a very subtle white-increasing effect, which is much less pronounced than the effect of the other described KIT variants. We termed the new KIT variants W18-W20 to provide a simple and unambiguous nomenclature for future genetic testing applications.

Hauswirth R; Jude R; Haase B; Bellone RR; Archer S; Holl H; Brooks SA; Tozaki T; Penedo MC; Rieder S; Leeb T

2013-05-01

263

Novel variants in the KIT and PAX3 genes in horses with white-spotted coat colour phenotypes.  

Science.gov (United States)

Variants in the EDNRB, KIT, MITF, PAX3 and TRPM1 genes are known to cause white spotting phenotypes in horses, which can range from the common white markings up to completely white horses. In this study, we investigated these candidate genes in 169 horses with white spotting phenotypes not explained by the previously described variants. We identified a novel missense variant, PAX3:p.Pro32Arg, in Appaloosa horses with a splashed white phenotype in addition to their leopard complex spotting patterns. We also found three novel variants in the KIT gene. The splice site variant c.1346+1G>A occurred in a Swiss Warmblood horse with a pronounced depigmentation phenotype. The missense variant p.Tyr441Cys was present in several part-bred Arabians with sabino-like depigmentation phenotypes. Finally, we provide evidence suggesting that the common and widely distributed KIT:p.Arg682His variant has a very subtle white-increasing effect, which is much less pronounced than the effect of the other described KIT variants. We termed the new KIT variants W18-W20 to provide a simple and unambiguous nomenclature for future genetic testing applications. PMID:23659293

Hauswirth, Regula; Jude, Rony; Haase, Bianca; Bellone, Rebecca R; Archer, Sheila; Holl, Heather; Brooks, Samantha A; Tozaki, Teruaki; Penedo, Maria Cecilia T; Rieder, Stefan; Leeb, Tosso

2013-05-01

264

Identification of a novel RNA silencing suppressor, NSs protein of Tomato spotted wilt virus.  

UK PubMed Central (United Kingdom)

RNA silencing or post-transcriptional gene silencing (PTGS) in plants is known as a defense system against virus infection. Several plant viruses have been shown to encode an RNA silencing suppressor. Using a green fluorescent protein-based transient suppression assay, we show that NSs protein of Tomato spotted wilt virus (TSWV) has RNA silencing suppressor activity. TSWV NSs protein suppressed sense transgene-induced PTGS but did not suppress inverted repeat transgene-induced PTGS. TSWV NSs protein is the first RNA silencing suppressor identified in negative-strand RNA viruses.

Takeda A; Sugiyama K; Nagano H; Mori M; Kaido M; Mise K; Tsuda S; Okuno T

2002-12-01

265

Cytopathological characteristics of tomato spotted wilt virus isolates  

Directory of Open Access Journals (Sweden)

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

Anna Rudzi?ska-Langwald; Maria Kami?ska

1998-01-01

266

The structure of melon necrotic spot virus determined at 2.8 A resolution.  

Science.gov (United States)

The structure of melon necrotic spot virus (MNSV) was determined at 2.8 A resolution. Although MNSV is classified into the genus Carmovirus of the family Tombusviridae, the three-dimensional structure of MNSV showed a higher degree of similarity to tomato bushy stunt virus (TBSV), which belongs to the genus Tombusvirus, than to carnation mottle virus (CMtV), turnip crinkle virus (TCV) or cowpea mottle virus (CPMtV) from the genus Carmovirus. Thus, the classification of the family Tombusviridae at the genus level conflicts with the patterns of similarity among coat-protein structures. MNSV is one of the viruses belonging to the genera Tombusvirus or Carmovirus that are naturally transmitted in the soil by zoospores of fungal vectors. The X-ray structure of MNSV provides us with a representative structure of viruses transmitted by fungi. PMID:18097092

Wada, Yasunobu; Tanaka, Hideaki; Yamashita, Eiki; Kubo, Chikako; Ichiki-Uehara, Tamaki; Nakazono-Nagaoka, Eiko; Omura, Toshihiro; Tsukihara, Tomitake

2007-12-20

267

Mice with mutations of Dock7 have generalized hypopigmentation and white-spotting but show normal neurological function  

Digital Repository Infrastructure Vision for European Research (DRIVER)

The classical recessive coat color mutation misty (m) arose spontaneously on the DBA/J background and causes generalized hypopigmentation and localized white-spotting in mice, with a lack of pigment on the belly, tail tip, and paws. Here we describe moonlight (mnlt), a second hypopigmentation and wh...

Blasius, Amanda L.; Brandl, Katharina; Crozat, Karine; Xia, Yu; Khovananth, Kevin; Krebs, Philippe; Smart, Nora G.

268

Alfalfa mosaic virus RNAs serve as cap donors for tomato spotted wilt virus transcription during coinfection of Nicotiana benthamiana.  

UK PubMed Central (United Kingdom)

Tomato spotted wilt virus (TSWV) was shown to use alfalfa mosaic virus (AMV) RNAs as cap donors in vivo during a mixed infection in Nicotiana benthamiana. By use of nested reverse transcription-PCR, TSWV N and NSs mRNAs provided with capped leader sequences derived from all four AMV RNAs could be cloned and sequenced. The sequence specificity of the putative TSWV endonuclease involved is discussed.

Duijsings D; Kormelink R; Goldbach R

1999-06-01

269

The nucleotide sequence of the S RNA of Impatiens necrotic spot virus, a novel tospovirus.  

UK PubMed Central (United Kingdom)

Impatiens necrotic spot virus (INSV) shares a number of properties with tomato spotted wilt virus (TSWV), the type species of the genus tospovirus within the family Bunyaviridae. INSV, however, differs from TSWV in plant host range and serology. In order to define the genomic structure and the taxonomic status of this TSWV-like virus, the nucleotide sequence of its genomic S RNA segment has been determined. The molecular data obtained demonstrate that, like TSWV, INSV has an ambisense S RNA molecule, encoding a non-structural protein in viral sense and the nucleocapsid protein in viral complementary sense. The level of nucleotide sequence homology between their S RNAs, as well as the divergence in amino acid sequence homology of their gene products, confirm previous conclusions from serological studies that INSV and TSWV represent distinct virus species within the newly created genus, tospovirus.

de Haan P; de Avila AC; Kormelink R; Westerbroek A; Gielen JJ; Peters D; Goldbach R

1992-07-01

270

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  

Scientific Electronic Library Online (English)

Full Text Available Abstract in portuguese 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 com (more) o 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). Abstract in english 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 (more) 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).

Calegario, Renata Faier; Ferreira, Sávio de Siqueira; Andrade, Eduardo Chumbinho de; Zerbini, Francisco Murilo

2007-09-01

271

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

Directory of Open Access Journals (Sweden)

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).

Renata Faier Calegario; Sávio de Siqueira Ferreira; Eduardo Chumbinho de Andrade; Francisco Murilo Zerbini

2007-01-01

272

Influenza A viruses in American White Pelican (Pelecanus erythrorhynchos).  

UK PubMed Central (United Kingdom)

The role of many wild waterbird species in the ecology and epidemiology of avian influenza viruses (AIV) remains unclear. We report the first isolation of AIV from American White Pelicans (Pelecanus erythrorhynchos; Pelecaniformes) in North America. Two H13N9 AIVs were isolated from hatchling birds in breeding colonies in Minnesota, USA, during 2007 and 2008. Based on molecular sequencing of the hemagglutinin and neuraminidase genes, the 2008 virus was genetically related to AIVs previously isolated from gulls and shorebirds in North America. The 2007 isolate was most related to AIVs from Eurasian gulls and North American ducks, reflecting both global movement of these viruses and reassortment between viruses associated with duck and gull reservoirs.

Lebarbenchon C; Sreevatsan S; Ramakrishnan MA; Poulson R; Goekjian V; Di Matteo JJ; Wilcox B; Stallknecht DE

2010-10-01

273

Influenza A viruses in American White Pelican (Pelecanus erythrorhynchos).  

Science.gov (United States)

The role of many wild waterbird species in the ecology and epidemiology of avian influenza viruses (AIV) remains unclear. We report the first isolation of AIV from American White Pelicans (Pelecanus erythrorhynchos; Pelecaniformes) in North America. Two H13N9 AIVs were isolated from hatchling birds in breeding colonies in Minnesota, USA, during 2007 and 2008. Based on molecular sequencing of the hemagglutinin and neuraminidase genes, the 2008 virus was genetically related to AIVs previously isolated from gulls and shorebirds in North America. The 2007 isolate was most related to AIVs from Eurasian gulls and North American ducks, reflecting both global movement of these viruses and reassortment between viruses associated with duck and gull reservoirs. PMID:20966281

Lebarbenchon, Camille; Sreevatsan, Srinand; Ramakrishnan, Muthannan A; Poulson, Rebecca; Goekjian, Virginia; Di Matteo, Jon J; Wilcox, Benjamin; Stallknecht, David E

2010-10-01

274

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

Scientific Electronic Library Online (English)

Full Text Available Abstract in spanish Se evaluó el efecto de ?-1,3-glucano, vitamina E y ?-caroteno en la dieta de juveniles de camarón café Farfantepenaeus californiensis inoculados con virus del síndrome de la mancha blanca (WSSV). Se colocaron grupos de 30 camarones (peso 1 ± 0,5 g) en contenedores de fibra de vidrio de 60 L y se alimentaron diariamente durante 23 días con ?-1,3-glucano (0,1%), vitamina E (0,01%), y ?-caroteno (0,01%) y posteriormente se inocularon con WSSV. Se de (more) terminó la actividad antioxidante de la enzima superóxido dismutasa (SOD) y catalasa (CAT) en hepatopáncreas y músculo a las 0, 1, 6, 12, 24 y 48 h después de la infección. Los grupos de camarones alimentados con los tratamientos incrementaron la actividad SOD en el hepatopáncreas y músculo a las 12 y 24 h después de la infección, respectivamente. Los juveniles tratados con vitamina E y ?-1,3-glucano mantuvieron un incremento en la actividad SOD desde las 12 a 48 h postinfección. Los camarones alimentados con ?-caroteno incrementaron la actividad de SOD antes de la infección con WSSV y los que fueron alimentados con ?-1,3-glucano y vitamina E incrementaron la actividad CAT también antes de la infección. La actividad CAT en músculo se incrementó respecto al grupo control, con todos los grupos de camarones tratados desde 1 hasta 6 h posteriores a la inoculación con WSSV. La actividad antioxidante más alta se registró en los camarones alimentados con vitamina E. Los juveniles alimentados con vitamina E y posteriormente inoculados con WSSV, registraron 100% de mortalidad a las 72 h, pero los que fueron alimentados con ?-1,3-glucano y ?-caroteno resistieron la infección hasta las 144 h. Los resultados de Antioxidant response in F. californiensis fed with dietary supplements and infected with WSSV este estudio mostraron la capacidad de juveniles de Farfantepenaeus californiensis alimentados con ?-1,3-glucano, vitamina E o ?-caroteno, de incrementar la respuesta antioxidante antes y durante una infección viral. Abstract in english The effect of dietary ?-?-glucan, vitamin E, and ?-carotene supplements in juvenile brown shrimp, Farfantepenaeus californiensis, inoculated with white spot syndrome virus (WSSV) was evaluated. Groups of 30 organisms (weighing 1 ± 0.5 g) were cultured in 60 L fiberglass tanks and fed daily with ?-1.3-glucan (0.1%), vitamin E (0.01%), and ?-carotene (0.01%) for 23 days; the specimens were then inoculated with WSSV. The antioxidant activity of the (more) enzymes superoxide dismutase (SOD) and catalase (CAT) were determined in the hepatopancreas and muscle at 0, 1, 6, 12, 24, and 48 h after inoculation. Shrimp fed with ?-1.3-glucan, vitamin E, and ?-carotene significantly increased SOD activity in the hepatopancreas and muscle at 12 and 24 h post-infection, respectively. Shrimp fed with vitamin E and ?-1.3-glucan registered an increment in SOD activity from 12 to 48 h post-infection. Shrimp fed with ?-carotene increased SOD activity before infection with WSSV, and shrimp fed with ?-1.3-glucan and vitamin E increased CAT activity, also before infection. The CAT activity response in shrimp muscle increased with respect to the control group for all treatments tested from 1 to 6 h after inoculation with WSSV. The highest antioxidant response was registered in shrimp fed with vitamin E. Juvenile shrimp fed with vitamin E and later inoculated with WSSV registered 100% mortality at 72 h, but shrimp fed with ?-?-glucan and ?-carotene showed greater resistance to WSSV, with mortality at 144 h post-infection. This study demonstrated the capacity of juvenile Farfantepenaeus californiensis fed ?-?-glucan, vitamin E, or ?-carotene to increase the antioxidant response before and after viral infection.

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

2011-11-01

275

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

Directory of Open Access Journals (Sweden)

Full Text Available The effect of dietary ?-?-glucan, vitamin E, and ?-carotene supplements in juvenile brown shrimp, Farfantepenaeus californiensis, inoculated with white spot syndrome virus (WSSV) was evaluated. Groups of 30 organisms (weighing 1 ± 0.5 g) were cultured in 60 L fiberglass tanks and fed daily with ?-1.3-glucan (0.1%), vitamin E (0.01%), and ?-carotene (0.01%) for 23 days; the specimens were then inoculated with WSSV. The antioxidant activity of the enzymes superoxide dismutase (SOD) and catalase (CAT) were determined in the hepatopancreas and muscle at 0, 1, 6, 12, 24, and 48 h after inoculation. Shrimp fed with ?-1.3-glucan, vitamin E, and ?-carotene significantly increased SOD activity in the hepatopancreas and muscle at 12 and 24 h post-infection, respectively. Shrimp fed with vitamin E and ?-1.3-glucan registered an increment in SOD activity from 12 to 48 h post-infection. Shrimp fed with ?-carotene increased SOD activity before infection with WSSV, and shrimp fed with ?-1.3-glucan and vitamin E increased CAT activity, also before infection. The CAT activity response in shrimp muscle increased with respect to the control group for all treatments tested from 1 to 6 h after inoculation with WSSV. The highest antioxidant response was registered in shrimp fed with vitamin E. Juvenile shrimp fed with vitamin E and later inoculated with WSSV registered 100% mortality at 72 h, but shrimp fed with ?-?-glucan and ?-carotene showed greater resistance to WSSV, with mortality at 144 h post-infection. This study demonstrated the capacity of juvenile Farfantepenaeus californiensis fed ?-?-glucan, vitamin E, or ?-carotene to increase the antioxidant response before and after viral infection.Se evaluó el efecto de ?-1,3-glucano, vitamina E y ?-caroteno en la dieta de juveniles de camarón café Farfantepenaeus californiensis inoculados con virus del síndrome de la mancha blanca (WSSV). Se colocaron grupos de 30 camarones (peso 1 ± 0,5 g) en contenedores de fibra de vidrio de 60 L y se alimentaron diariamente durante 23 días con ?-1,3-glucano (0,1%), vitamina E (0,01%), y ?-caroteno (0,01%) y posteriormente se inocularon con WSSV. Se determinó la actividad antioxidante de la enzima superóxido dismutasa (SOD) y catalasa (CAT) en hepatopáncreas y músculo a las 0, 1, 6, 12, 24 y 48 h después de la infección. Los grupos de camarones alimentados con los tratamientos incrementaron la actividad SOD en el hepatopáncreas y músculo a las 12 y 24 h después de la infección, respectivamente. Los juveniles tratados con vitamina E y ?-1,3-glucano mantuvieron un incremento en la actividad SOD desde las 12 a 48 h postinfección. Los camarones alimentados con ?-caroteno incrementaron la actividad de SOD antes de la infección con WSSV y los que fueron alimentados con ?-1,3-glucano y vitamina E incrementaron la actividad CAT también antes de la infección. La actividad CAT en músculo se incrementó respecto al grupo control, con todos los grupos de camarones tratados desde 1 hasta 6 h posteriores a la inoculación con WSSV. La actividad antioxidante más alta se registró en los camarones alimentados con vitamina E. Los juveniles alimentados con vitamina E y posteriormente inoculados con WSSV, registraron 100% de mortalidad a las 72 h, pero los que fueron alimentados con ?-1,3-glucano y ?-caroteno resistieron la infección hasta las 144 h. Los resultados de Antioxidant response in F. californiensis fed with dietary supplements and infected with WSSV este estudio mostraron la capacidad de juveniles de Farfantepenaeus californiensis alimentados con ?-1,3-glucano, vitamina E o ?-caroteno, de incrementar la respuesta antioxidante antes y durante una infección viral.

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

2011-01-01

276

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

Digital Repository Infrastructure Vision for European Research (DRIVER)

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

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

277

UTILIZING IMMUNOASSAYS TO DETERMINE SYSTEMIC TOMATO SPOTTED WILT VIRUS INFECTION FOR ELUCIDATING FIELD RESISTANCE IN PEANUT  

Science.gov (United States)

Three cultivars, SunOleic95R, Southern Runner, and DP-1, were evaluated for tomato spotted wilt virus (TSWV) resistance in replicated field plots at Marianna, FL, in 1998 and 1999. Treatments consisted of April and May planting dates with 7.6 or 15.2 cm space plantings. The presence of TSWV was de...

278

Resistance to tomato spotted wilt virus and root-knot nematode in peanut interspecific breeding lines  

Science.gov (United States)

The peanut root-knot nematode [Meloidogyne arenaria (Neal) Chitwood race 1] and tomato spotted wilt virus Tospovirus (TSWV) are economically significant pathogens of peanut in the southeastern United States. Peanut cultivars are available that have resistance to either the peanut root-knot nematode...

279

Engineered RNA-mediated resistance to tomato spotted wilt virus is sequence specific.  

UK PubMed Central (United Kingdom)

Transgenic plants were produced that expressed a wide range of randomly chosen sequences of the tripartite tomato spotted wilt virus (TSWV) RNA genome or its complement. Testing the progenies of these plants revealed that only transgenic expression of N or NS(M) gene sequences resulted in resistance to TSWV.

Prins M; Resende Rde O; Anker C; van Schepen A; de Haan P; Goldbach R

1996-07-01

280

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

Science.gov (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...

 
 
 
 
281

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

Digital Repository Infrastructure Vision for European Research (DRIVER)

Black root rot (BRR) caused by Thielaviopsis basicola as well as Tomato spotted wilt virus (TSWV) are the most serious problems in tobacco growing regions. We crossed the breeding line WGL 3 carrying BRR resistance derived from N.glauca with the line PW-834 the resistance of which to TSWV was transf...

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

282

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

UK PubMed Central (United Kingdom)

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

Culbreath AK; Srinivasan R

2011-08-01

283

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

Science.gov (United States)

Spotted wilt disease of peanut (Arachis hypogaea) (SWP), caused by Tomato spotted wilt virus (TSWV) (genus Tospovirus, family Bunyaviridae), was first observed in Alabama, Florida, and Georgia in the late 1980s and rapidly became a major limiting factor for peanut production in the region. Tobacco thrips (Frankliniella fusca) and western flower thrips (Frankliniella occidentalis) both occur on peanut throughout the southeastern U.S., but F. fusca is the predominant species that reproduces on peanut, and is considered to be the more important vector. Several non-crop sources of potential primary vectors and TSWV inoculum have been identified, but their relative importance has not been determined. The peanut growing season in Alabama, Florida, and Georgia is from April through November, and 'volunteer' peanut plants can be present for much of the remainder of the year. Therefore peanut itself has huge potential for perpetuating both vector and virus. Symptoms are often evident within a few days of seedling emergence, and disease progress is often rapid within the first 50-60 days after planting. Based on destructive sampling and assays for TSWV, there is often a high incidence of asymptomatic infections even in peanut genotypes that produce few and mild symptoms of infection in the field. Severity of SWP epidemics fluctuates significantly from year to year. The variability has not been fully explained, but lower incidences have been associated with years categorized as "La Niña" in the El Niño-Southern Oscillation. Planting date can have a large effect on disease incidence within a location. This may be linked to the thrips reproductive cycle and environmental effects on the plant and plant-thrips-virus interactions. Row pattern, plant population, and in-furrow applications of phorate insecticide can also affect epidemics of SWP. Considerable progress has been made in developing cultivars with natural field resistance to TSWV. Use of cultivars with moderate field resistance combined with other suppressive measures has been very successful for managing spotted wilt disease. Several new cultivars with higher levels of field resistance can improve control and allow more flexibility in the integrated management programme. Although effects of these factors on epidemics of SWP have been documented, mechanisms responsible for disease suppression by most factors have not been fully elucidated. PMID:21620508

Culbreath, A K; Srinivasan, R

2011-04-28

284

Tissue distribution of metals in white-fronted geese and spot-billed ducks from Korea.  

UK PubMed Central (United Kingdom)

This study presents concentrations of Fe, Zn, Mn, Cu, Pb and Cd in livers, kidneys, muscles and bones of white-fronted geese Anser albifrons (geese) and spot-billed ducks Anas poecilorhyncha (ducks). Iron in livers, kidneys and muscles, Zn in muscles, Mn and Cd in every tissue, Cu in livers, muscles and bones and Pb in bones differed between species, and there were significant differences among tissues in both species. Essential elements such as Fe, Zn, Mn and Cu concentrations were within the background levels. Lead concentrations in livers of 7 of 14 geese and 7 of 19 ducks and in bones of 4 of 19 ducks exceeded background concentrations for waterfowl (5 ?g/g dw for the liver, 10 ?g/g dw for the bone). Almost all samples of both species had the background Cd concentrations in the liver (33 of 33 geese and ducks) and kidney (14 geese and 18 ducks). Tissue concentrations of Cd were greater in geese than ducks. In contrast, tissue concentrations of Pb in bones were greater in ducks than in geese. These different trends for Cd and Pb reflect a short and/or long term difference in exposure and degree of accumulation of these metals.

Kim J; Oh JM

2013-07-01

285

Esthetic comparison of white-spot lesion treatment modalities using spectrometry and fluorescence.  

UK PubMed Central (United Kingdom)

Abstract Objective: To compare the esthetic improvements of white-spot lesions (WSLs) treated by fluoride, casein phosphopeptide amorphous calcium phosphate (CPP-ACP), or resin infiltration. Materials and Methods: WSLs were created on human enamel and randomly assigned to four groups: NaF (500 ppm), CPP-ACP, resin infiltration (Icon), or distilled deionized water (DDW; control group). The color change (?E) of each specimen was measured with a Crystaleye spectrophotometer, and fluorescence loss (?Q) was measured by quantitative light-induced fluorescence (QLF), at different time points after treatment: baseline (0 weeks), 2 weeks, 4 weeks, and 6 weeks. Results: The ?E and ?Q baseline values for the four groups before the treatments did not differ significantly. Icon treatment improved the WSL color significantly and gave the lowest ?E (2.9 ± 1.2 on average) compared with other treatments (P < .01). The Icon treatment also resulted in a significant change in the ?Q of WSLs compared with baseline (P < .01). In the NaF and CPP-ACP treatment groups, ?Q showed significant recovery compared with the baseline values only after 4 weeks after treatment (P < .05). Conclusions: Resin infiltration is more effective than NaF or CPP-ACP in providing esthetic improvement of WSLs.

Yuan H; Li J; Chen L; Cheng L; Cannon RD; Mei L

2013-08-01

286

The evaluation of resin infiltration for masking labial enamel white spot lesions.  

UK PubMed Central (United Kingdom)

OBJECTIVE: The aim of this study was to clinically assess the effectiveness of masking white spot enamel lesions using a resin infiltration technique that was recently developed to arrest incipient caries in a micro-invasive concept. METHODS: Twenty teeth with a Developmental Defect of Enamel (DDE) and 18 teeth with Post-orthodontic Decalcification (POD) were selected and treated with resin infiltration. Standardized photographs were taken before, immediately after, and 1 week after treatment and were analysed using image analysing software to calculate the ?E values. The results were classified into three groups: completely masked, partially masked, and unchanged. RESULTS: Among the 20 teeth with DDE, five teeth (25%) were classified as completely masked, whereas seven (35%) and eight teeth (40%) were partially masked and unchanged, respectively. Among the 18 teeth with POD, 11 teeth (61%) were completely masked, six teeth (33%) were partially masked, and one tooth (6%) was unchanged. In some teeth, the result was more improved after 1 week than immediately after infiltration. CONCLUSION: The masking effect was dramatic in some cases but not in others. The long-term colour stability of the result should be followed up through continuous clinical and scientific studies.

Kim S; Kim EY; Jeong TS; Kim JW

2011-07-01

287

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

Science.gov (United States)

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 < 0.05). Intraobserver agreement was assessed by means of the Cohen ? statistical method. There were no significant differences in the prevalence of WSLs between patients treated for 6 and 12 months (P = 0.855); however, there were significantly more WSLs in groups I and II than in group 0 (P = 0.000). No significant differences were found between girls and boys (P = 1.000). The mandibular first molars and maxillary lateral incisors were the most affected teeth, in both the treated and untreated groups. The study revealed significant decalcification at 6 months after orthodontic bonding. Considering how quickly these lesions can develop and become irreversible, early diagnosis is of critical importance. PMID:23045306

Lucchese, Alessandra; Gherlone, Enrico

2012-10-08

288

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

Directory of Open Access Journals (Sweden)

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

Jainara Maria Soares Ferreira; Ana Karla Ramalho Aragão; Adriana Dias Batista Rosa; Fábio Correia Sampaio; Valdenice Aparecida de Menezes

2009-01-01

289

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

Scientific Electronic Library Online (English)

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

Ferreira, Jainara Maria Soares; Aragão, Ana Karla Ramalho; Rosa, Adriana Dias Batista; Sampaio, Fábio Correia; Menezes, Valdenice Aparecida de

2009-12-01

290

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

UK PubMed Central (United Kingdom)

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 < 0.05). Intraobserver agreement was assessed by means of the Cohen ? statistical method. There were no significant differences in the prevalence of WSLs between patients treated for 6 and 12 months (P = 0.855); however, there were significantly more WSLs in groups I and II than in group 0 (P = 0.000). No significant differences were found between girls and boys (P = 1.000). The mandibular first molars and maxillary lateral incisors were the most affected teeth, in both the treated and untreated groups. The study revealed significant decalcification at 6 months after orthodontic bonding. Considering how quickly these lesions can develop and become irreversible, early diagnosis is of critical importance.

Lucchese A; Gherlone E

2013-10-01

291

Potential role of viruses in white plague coral disease.  

UK PubMed Central (United Kingdom)

White plague (WP)-like diseases of tropical corals are implicated in reef decline worldwide, although their etiological cause is generally unknown. Studies thus far have focused on bacterial or eukaryotic pathogens as the source of these diseases; no studies have examined the role of viruses. Using a combination of transmission electron microscopy (TEM) and 454 pyrosequencing, we compared 24 viral metagenomes generated from Montastraea annularis corals showing signs of WP-like disease and/or bleaching, control conspecific corals, and adjacent seawater. TEM was used for visual inspection of diseased coral tissue. No bacteria were visually identified within diseased coral tissues, but viral particles and sequence similarities to eukaryotic circular Rep-encoding single-stranded DNA viruses and their associated satellites (SCSDVs) were abundant in WP diseased tissues. In contrast, sequence similarities to SCSDVs were not found in any healthy coral tissues, suggesting SCSDVs might have a role in WP disease. Furthermore, Herpesviridae gene signatures dominated healthy tissues, corroborating reports that herpes-like viruses infect all corals. Nucleocytoplasmic large DNA virus (NCLDV) sequences, similar to those recently identified in cultures of Symbiodinium (the algal symbionts of corals), were most common in bleached corals. This finding further implicates that these NCLDV viruses may have a role in bleaching, as suggested in previous studies. This study determined that a specific group of viruses is associated with diseased Caribbean corals and highlights the potential for viral disease in regional coral reef decline.

Soffer N; Brandt ME; Correa AM; Smith TB; Thurber RV

2013-08-01

292

Effectiveness of pit and fissure sealants in reducing white spot lesions during orthodontic treatment. A pilot study.  

UK PubMed Central (United Kingdom)

OBJECTIVE: A pilot investigation was performed to test the null hypothesis that highly filled (58%) resin sealants do not prevent white spot lesions in patients undergoing active orthodontic treatment. MATERIALS AND METHODS: A split-mouth design was applied to 60 healthy patients, with the sealant randomly allocated to either the right or the left side of each jaw. The sealant was applied to the incisors and canines from the gingival surface of the bracket to the free gingival margin. The contralateral teeth had the same type of bracket with no sealant. Sealants were placed on the experimental teeth 2 weeks to 3 months after initial bonding and were removed after 15 to 18 months. Intraoral photographs, visual assessments, and DIAGNOdent (KaVo Dental Corporation, Lake Zurich, Ill) measurements were used to assess white spot lesions after sealant removal. RESULTS: Six lesions on the teeth with sealants were identified visually, compared with 22 lesions on the teeth without sealants. The teeth without sealants had 3.8 times the number of white spot lesions than were noted on the sealed teeth. These sealants showed no visible signs of discoloration. The DIAGNOdent measured statistically significant differences between sealed and unsealed teeth in the maxilla (P < .001) and in the mandible (P = .010). DIAGNOdent measurements also showed a difference between sealed and unsealed teeth after the 28 teeth with visible lesions were excluded. CONCLUSION: The hypothesis was rejected. Ultraseal XT Plus clear sealant (Ultradent Products, South Jordon, Utah) produced a significant reduction in enamel demineralization during fixed orthodontic treatment and should be considered for use by clinicians to minimize white spot lesions.

Benham AW; Campbell PM; Buschang PH

2009-03-01

293

Effect of 1.23% acidulated phosphate fluoride foam on white spot lesions in orthodontic patients: a randomized trial.  

UK PubMed Central (United Kingdom)

PURPOSE: The decalcification of enamel is a serious clinical problem in orthodontic patients and is usually observed as white spot lesions surrounding brackets. This study's purpose was to evaluate the effect of professionally applied 1.23 percent fluoride foam on reducing the formation of white spot lesions (WSLs) in patients with fixed orthodontic appliances. METHODS: In a randomized, double-blind, placebo-controlled trial, 100 participants were randomly divided into two groups. The two groups received fluoride foam and placebo foam, respectively, every two months during the treatment. The examinations before bonding and after debonding were performed by one examiner and included the presence and severity of WSLs on incisors, canines, and premolars. RESULTS: The incidence of WSLs was approximately 13 percent in the fluoride foam group and 51 percent in the placebo group (P<.001). The mean increment of WSLs score was significantly lower in the fluoride foam group (0.71 ± 2.80) than in the placebo group (4.36 ± 5.41; P<.001). The preventive fraction was approximately 76 percent, and the number needed to treat was calculated as 2.6. CONCLUSIONS: Professional application of 1.23 percent fluoride foam during orthodontic treatment effectively reduced the development of white spot lesions. A prophylactic regimen based on the routine use of fluoride foam during orthodontic treatment is recommended.

Jiang H; Hua F; Yao L; Tai B; Du M

2013-05-01

294

Herpesviruses and Newcastle disease viruses in white storks (Ciconia ciconia).  

Science.gov (United States)

Three herpesviruses were isolated from white storks (Ciconia ciconia). All isolates reacted in cross-neutralisation tests with homologous antisera and with sera prepared against a herpesvirus from a black stork (Ciconia nigra). These data indicate serologic relatedness of the herpesviruses from both stork species. Antisera prepared against herpesviruses from the domestic chicken (viruses of Marek's disease and infectious laryngotracheitis), turkey, duck and pigeon as well as from the blue-fronted amazon (Amazona aestiva), prairie falcon (Falco mexicanus), eagle owl (Bubo bubo), Lake Victoria cormorant (Phalacrocorax melanoleucos), bobwhite quail (Colinus virginianus) and desmoiselle crane (Anthropoides virgo) did not react with the stork herpesviruses. Neutralising antibodies against stork herpesvirus were detected in the majority of 72 blood samples from white and black storks. In addition, three Newcastle disease viruses (NDV) could be isolated from white storks. One isolate was highly virulent the two others were avirulent for the chicken. Haemagglutination inhibition tests have shown that some storks have antibodies against Paramyxovirus- (PMV)-1 (NDV), PMV-2 and PMV-3. No antibodies could be detected in stork sera against PMV-4, -6 and -7. PMID:18766791

Kaleta, E F; Kummerfeld, N

1983-01-01

295

Herpesviruses and Newcastle disease viruses in white storks (Ciconia ciconia).  

UK PubMed Central (United Kingdom)

Three herpesviruses were isolated from white storks (Ciconia ciconia). All isolates reacted in cross-neutralisation tests with homologous antisera and with sera prepared against a herpesvirus from a black stork (Ciconia nigra). These data indicate serologic relatedness of the herpesviruses from both stork species. Antisera prepared against herpesviruses from the domestic chicken (viruses of Marek's disease and infectious laryngotracheitis), turkey, duck and pigeon as well as from the blue-fronted amazon (Amazona aestiva), prairie falcon (Falco mexicanus), eagle owl (Bubo bubo), Lake Victoria cormorant (Phalacrocorax melanoleucos), bobwhite quail (Colinus virginianus) and desmoiselle crane (Anthropoides virgo) did not react with the stork herpesviruses. Neutralising antibodies against stork herpesvirus were detected in the majority of 72 blood samples from white and black storks. In addition, three Newcastle disease viruses (NDV) could be isolated from white storks. One isolate was highly virulent the two others were avirulent for the chicken. Haemagglutination inhibition tests have shown that some storks have antibodies against Paramyxovirus- (PMV)-1 (NDV), PMV-2 and PMV-3. No antibodies could be detected in stork sera against PMV-4, -6 and -7.

Kaleta EF; Kummerfeld N

1983-01-01

296

The effect of fractional CO2 laser irradiation on remineralization of enamel white spot lesions.  

UK PubMed Central (United Kingdom)

This study investigated the combined effect of fractional CO2 laser irradiation and fluoride on treatment of enamel caries. Sixty intact premolars were randomly assigned into four groups and then stored in a demineralizing solution to induce white spot lesions. Tooth color was determined at baseline (T1) and after demineralization (T2). Afterwards, the teeth in group 1 remained untreated (control), while group 2 was exposed to an acidulated phosphate fluoride (APF) gel for 4 min. In groups 3 and 4, a fractional CO2 laser was applied (10 mJ, 200 Hz, 10 s) either before (group 3) or through (group 4) the APF gel. The teeth were then immersed in artificial saliva for 90 days while subjected to daily fluoride mouthrinse and weekly brushing. Color examinations were repeated after topical fluoride application (T3) and 90 days later (T4). Finally, the teeth were sectioned, and microhardness was measured at the enamel surface and at 30 and 60 ? from the surface. In both lased groups, the color change between T1 and T4 stages (?E T1-T4) was significantly lower than those of the other groups (p?

Poosti M; Ahrari F; Moosavi H; Najjaran H

2013-03-01

297

Performance of iron spot test with Arabic bread made from fortified white wheat flour.  

UK PubMed Central (United Kingdom)

BACKGROUND: The iron spot test (IST) is a simple qualitative technique for determining the presence of added iron in fortified flour. IST performance in bread has never been investigated. If found to perform well, the IST has the potential to provide a field-friendly method for testing bread and thus support the monitoring and evaluation of flour fortification programs. OBJECTIVE: To assess the performance of the IST in Arabic bread made from white wheat flour. METHODS: Bread samples were collected from 1,737 households during a national micronutrient survey in Jordan. A subsample of Arabic bread (n = 44) was systematically selected for testing by both the IST and spectrophotometry (criterion reference). Performance measures (sensitivity, specificity, and positive and negative predictive values) were calculated using five cutoffs to define the presence of added iron, including > or = 15.0 ppm (approximate level of natural iron in Arabic bread) and four additional cutoffs based on test performance. RESULTS: The iron contents of samples testing negative by IST ranged from 10.4 to 18.4 ppm, with one outlier at 41.0 ppm, which was excluded from subsequent analyses. The iron contents of samples testing positive by IST ranged from 16.1 to 38.4 ppm. With the exception of negative predictive values for the two lowest cutoffs (> or = 15.0 and > or = 16.1 ppm), all performance measures exceeded 83.3%. CONCLUSIONS: These results show promise for the IST as an inexpensive, field-friendly method for testing bread that could have a useful role in the monitoring and evaluation process for flour fortification programs.

Nichols E; Aburto N; Masa'd H; Wirth J; Sullivan K; Serdula M

2012-09-01

298

Micro-spot with integrated pillars (MSIP) for detection of dengue virus NS1.  

UK PubMed Central (United Kingdom)

In this paper, we demonstrate an extremely efficient technique of diagnosing dengue virus non-structural protein (NS1) using Micro-Spot with Integrated Pillars (MSIP). Detection using MSIP is performed by employing fluorescence immunoassay specific to dengue virus NS1. MSIPs are chemically modified to ensure efficient covalent binding of antibodies on the micropillars, whereas the enormous increase in the surface area (available for the reaction) induced by the micropillars amplifies the apparent rate, which enhances the signal intensity. Therefore, the detection response of a MSIP, quantified by the intensity of the fluorescence signal, is found to be almost five times magnified than the response of a similar size micro-spot without micropillars. The response of the micropillars also depend on the pillar arrangement, since for identical concentration of dengue NS1 antigen, a stronger intensity signal is obtained for a hexagonal close packed array (staggered) pillar arrangement as compared to a square array arrangement.

Gunda NS; Singh M; Purwar Y; Shah SL; Kaur K; Mitra SK

2013-07-01

299

Tomato spotted wilt virus nucleocapsid protein interacts with both viral glycoproteins Gn and Gc in planta.  

UK PubMed Central (United Kingdom)

Recently, the Tomato Spotted Wilt Virus (TSWV) Gn and Gc glycoproteins were shown to induce the formation of (pseudo-) circular and pleomorphic membrane structures upon transient expression in plant cells. Furthermore, when singly expressed, Gc retains in the ER, while Gn is able to further migrate to the Golgi. Upon co-expression, Gn rescues Gc and co-migrates to the Golgi complex. Here, we have studied the behavior of the glycoproteins in the presence of the viral nucleocapsid (N) protein and in vivo analyzed the occurrence of protein-protein interactions by fluorescence life time imaging microscopy (FLIM). The analysis demonstrated that N co-localizes and interacts with both glycoproteins, with a preference for Gn. Additionally, it is shown that N causes a dramatic change in the distribution of Gc within the ER, from reticular to punctate spots. The observations are discussed in the context of the virus particle formation during the infection process.

Ribeiro D; Borst JW; Goldbach R; Kormelink R

2009-01-01

300

Tomato spotted wilt virus nucleocapsid protein interacts with both viral glycoproteins Gn and Gc in planta.  

Science.gov (United States)

Recently, the Tomato Spotted Wilt Virus (TSWV) Gn and Gc glycoproteins were shown to induce the formation of (pseudo-) circular and pleomorphic membrane structures upon transient expression in plant cells. Furthermore, when singly expressed, Gc retains in the ER, while Gn is able to further migrate to the Golgi. Upon co-expression, Gn rescues Gc and co-migrates to the Golgi complex. Here, we have studied the behavior of the glycoproteins in the presence of the viral nucleocapsid (N) protein and in vivo analyzed the occurrence of protein-protein interactions by fluorescence life time imaging microscopy (FLIM). The analysis demonstrated that N co-localizes and interacts with both glycoproteins, with a preference for Gn. Additionally, it is shown that N causes a dramatic change in the distribution of Gc within the ER, from reticular to punctate spots. The observations are discussed in the context of the virus particle formation during the infection process. PMID:18973913

Ribeiro, Daniela; Borst, Jan Willem; Goldbach, Rob; Kormelink, Richard

2008-10-29

 
 
 
 
301

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

UK PubMed Central (United Kingdom)

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

Caner J; Amélia M; Alexandre V; Vicente M

1984-12-01

302

Acquisition of Tomato spotted wilt virus by Adults of Two Thrips Species.  

UK PubMed Central (United Kingdom)

ABSTRACT Only larval thrips that acquire Tomato spotted wilt virus (TSWV), or adults derived from such larvae, transmit the virus. Nonviruliferous adults can ingest virus particles while feeding on TSWV-infected plants, but such adult thrips have not been shown to transmit TSWV. Immunofluorescence microscopy was used to show that thrips 1, 5, 10, and 20 days after adult emergence (DAE) fed on TSWV-infected plants acquired TSWV with virus replication and accumulation occurring in both epithelial and muscle cells of Frankliniella fusca (tobacco thrips [TT]) and F. occidentalis (western flower thrips [WFT]), as indicated by immunodetection of the nonstructural (NSs) protein encoded by the small RNA and the nucleocapsid (N) protein, respectively. Adult WFT acquired TSWV more efficiently than TT. There was no significant effect of insect age on TSWV acquisition by TT. In contrast, acquisition by adult WFT at 1 and 5 DAE was higher than acquisition at 10 and 20 DAE. Subsequent transmission competence of adult cohorts was studied by vector transmission assays. All adult thrips tested that had an acquisition access period as an adult were unable to transmit the virus. These results indicate the susceptibility of adult TT and WFT to infection of midgut cells by TSWV and subsequent virus replication and confirm earlier studies that adult thrips that feed on virus-infected plants do not transmit the virus. The role of a tissue barrier in TSWV movement and infection from midgut muscle cells to the salivary glands is discussed.

de Assis Filho FM; Deom CM; Sherwood JL

2004-04-01

303

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

UK PubMed Central (United Kingdom)

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.

Torres R; Larenas J; Fribourg C; Romero J

2012-04-01

304

Evaluation of 'white-spotted kidneys' associated with leptospirosis by polymerase chain reaction based LipL32 gene in slaughtered cows  

Scientific Electronic Library Online (English)

Full Text Available Abstract in english The presence of white spots in the kidneys of cattle at slaughter (so-called white-spotted kidneys) can be an indication of infection with Leptospira, a spirochaete of public health concern because it causes zoonotic disease. In this study, 24 kidneys of 180 slaughtered cows (13.3%) showed focal to multifocal white spots at inspection. These kidneys, together with matching urine (n = 18) and blood (n = 24) samples, were examined by polymerase chain reaction (PCR) targetin (more) g the LipL32 gene. Leptospiral deoxyribonucleic acid (DNA) was detected in 19 (79.2%) out of 24 kidneys, as well as 7 (29.2%) blood and 10 (55.5%) urine samples of cows with white spots in their kidneys. Histopathological findings revealed multifocal infiltration of mononuclear cells, including lymphocytes and a few plasma cells in the renal interstitial tissues. In addition, 14 apparently normal kidneys and associated urine and blood samples were similarly examined by PCR but did not provide any positive results. In this study, high detection of leptospirosis in kidneys with interstitial nephritis suggests that Leptospira spp. are associated with white spotted kidneys. The present findings indicate that white spotted kidneys can be due to leptospirosis in this region in southwestern Iran, which indicates an increased risk of zoonotic disease. The data show that LipL32-based primers are useful for PCR-based diagnosis of leptospirosis.

Azizi, Shahrzad; Tajbakhsh, Elahe; Hajimirzaei, Mohammad R.; Varnamkhasti, Mohssen Gholami; Sadeghian, Hossein; Oryan, Ahmad

2012-01-01

305

Evaluation of ‘white-spotted kidneys’ associated with leptospirosis by polymerase chain reaction based LipL32 gene in slaughtered cows  

Directory of Open Access Journals (Sweden)

Full Text Available The presence of white spots in the kidneys of cattle at slaughter (so-called white-spotted kidneys) can be an indication of infection with Leptospira, a spirochaete of public health concern because it causes zoonotic disease. In this study, 24 kidneys of 180 slaughtered cows (13.3%) showed focal to multifocal white spots at inspection. These kidneys, together with matching urine (n = 18) and blood (n = 24) samples, were examined by polymerase chain reaction (PCR) targeting the LipL32 gene. Leptospiral deoxyribonucleic acid (DNA) was detected in 19 (79.2%) out of 24 kidneys, as well as 7 (29.2%) blood and 10 (55.5%) urine samples of cows with white spots in their kidneys. Histopathological findings revealed multifocal infiltration of mononuclear cells, including lymphocytes and a few plasma cells in the renal interstitial tissues. In addition, 14 apparently normal kidneys and associated urine and blood samples were similarly examined by PCR but did not provide any positive results. In this study, high detection of leptospirosis in kidneys with interstitial nephritis suggests that Leptospira spp. are associated with white spotted kidneys. The present findings indicate that white spotted kidneys can be due to leptospirosis in this region in southwestern Iran, which indicates an increased risk of zoonotic disease. The data show that LipL32-based primers are useful for PCR-based diagnosis of leptospirosis.

Shahrzad Azizi; Elahe Tajbakhsh; Mohammad R. Hajimirzaei; Mohssen Gholami Varnamkhast; Hossein Sadeghian; Ahmad Oryan

2012-01-01

306

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

Directory of Open Access Journals (Sweden)

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

Zaharova E; Sherman J

2011-01-01

307

The molecular population genetics of the Tomato spotted wilt virus (TSWV) genome.  

Science.gov (United States)

RNA viruses are characterized by high genetic variability resulting in rapid adaptation to new or resistant hosts. Research for plant RNA virus genetic structure and its variability has been relatively scarce compared to abundant research done for human and animal RNA viruses. Here, we utilized a molecular population genetic framework to characterize the evolution of a highly pathogenic plant RNA virus [Tomato spotted wilt virus (TSWV), Tospovirus, Bunyaviridae]. Data from genes encoding five viral proteins were used for phylogenetic analysis, and for estimation of population parameters, subpopulation differentiation, recombination, divergence between Tospovirus species, and selective constraints on the TSWV genome. Our analysis has defined the geographical structure of TSWV, attributed possibly to founder effects. Also, we identify positive selection favouring divergence between Tospovirus species. At the species level, purifying selection has acted to preserve protein function, although certain amino acids appear to be under positive selection. This analysis provides demonstration of population structuring and species-wide population expansions in a multisegmented plant RNA virus, using sequence-based molecular population genetic analyses. It also identifies specific amino acid sites subject to selection within Bunyaviridae and estimates the level of genetic heterogeneity of a highly pathogenic plant RNA virus. The study of the variability of TSWV populations lays the foundation in the development of strategies for the control of other viral diseases in floral crops. PMID:15643950

Tsompana, M; Abad, J; Purugganan, M; Moyer, J W

2005-01-01

308

Tomato spotted wilt virus infection reduces the fitness of a nonvector herbivore on pepper.  

Science.gov (United States)

Plant pathogens and insect herbivores often share hosts under natural conditions. Hence, pathogen-induced changes in a host plant can affect the herbivore and vice versa. Even though plant viruses are ubiquitous in the field, little is known about plant-mediated interactions between viruses and nonvector herbivores. Here we tested whether the performance of the sweet potato whitefly, Bemisia tabaci (Gennadius) biotype Q, was altered when raised on pepper infected with Tomato spotted wilt virus (TSWV). TSWV infection reduced B. tabaci fecundity and longevity and increased B. tabaci developmental time but did not affect the insect's survival or female body lengths. Our results demonstrate that TSWV infection can decrease the fitness of B. tabaci biotype Q on pepper plants. PMID:23786083

Pan, Huipeng; Chen, Gong; Li, Fei; Wu, Qingjun; Wang, Shaoli; Xie, Wen; Liu, Baiming; Xu, Baoyun; Zhang, Youjun

2013-04-01

309

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

UK PubMed Central (United Kingdom)

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

Czech AS; Szklarczyk M; Gajewski Z; Zukowska E; Michalik B; Koby?ko T; Strza?ka K

2003-01-01

310

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

Scientific Electronic Library Online (English)

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

Velásquez-Valle, Rodolfo; Reveles-Hernández, Manuel

2011-12-01

311

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

Digital Repository Infrastructure Vision for European Research (DRIVER)

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 ...

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

312

Differential responses of selected peanut cultivars and breeding lines to mechanical inoculation of tomato spotted wilt virus  

Science.gov (United States)

Screening of peanut germplasm for resistance to Tomato spotted wilt virus (TSWV) has been largely inefficient due to the lack of a screening technique based on mechanical transmission of the virus under controlled environmental conditions. We have studied the reaction of three peanut cultivars (Geor...

313

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

Directory of Open Access Journals (Sweden)

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

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

2012-01-01

314

Passion fruit green spot virus vectored by Brevipalpus phoenicis (Acari: Tenuipalpidae) on passion fruit in Brazil.  

Science.gov (United States)

Passion fruit green spot disease was first identified in 1997 after a severe outbreak at Vera Cruz County, state of São Paulo, Brazil. Mature yellow fruits of Passiflora edulis Simms f. flavicarpa Degener showed characteristic green spots, 2-5 mm in diameter and patches of green tissues were present on senescent leaves. The devastating effect to passion flower is caused by necrotic lesions that encircle the stems and kill the plant. In severe cases, entire orchards of a few hectares in size have been completely destroyed. The disease was always preceded by heavy infestations of Brevipalpus phoenicis (Geijskes) (Acari: Tenuipalpidae). Transmission electron microscopy of affected tissues (fruits, leaves, and stems) consistently revealed the presence of short, bacilliform particles (50-70 nm x 100-120 nm) in the cisternae of the endoplasmic reticulum, as well as the presence of a dense viroplasm in the cytoplasm. This cytopathic effect has been found in several other Brevipalpus-transmitted or associated viruses and is classified as a cytoplasmic type of disease. Experimental reproduction of the leaf and stem symptoms was achieved by transferring B. phoenicis collected from affected field passion flower plants onto healthy plants. The evidence supports a viral etiology for the disease and the agent was named passion fruit green spot virus. Its relationship with other B. phoenicis related viruses continues to be studied. The disease was also found in the Brazilian states of Bahia, Sergipe, Rondonia, Minas Gerais, Rio de Janeiro, and in the Federal District. Use of one or more of the following acaricides (hexythiazox, fenbutatin-oxide, propargite, quinomethionate, or dicofol) has significantly reduced the incidence of the disease. PMID:14756419

Kitajima, E W; Rezende, J A M; Rodrigues, J C V

2003-01-01

315

Melon yellow spot virus: A Distinct Species of the Genus Tospovirus Isolated from Melon.  

UK PubMed Central (United Kingdom)

ABSTRACT A tospovirus-like virus recovered from netted melon was transmitted by Thrips palmi in a persistent manner but had different cytopathological features from tospoviruses previously reported. Viral nucleocapsid (N) was purified with two protective reagents, 2-mercaptoethanol and L-ascorbic acid, and RNA extracted from the viral nucleocapsid was used for genomic analysis. The virus had a genome consisting of three single-stranded RNA molecules. The open reading frame on the viral complementary strand, located at the 3' end of the viral S RNA, encoded the N protein. The 3' terminus of this RNA also contained an eight-nucleotide sequence similar to the conserved sequence at the 3' end of genomic RNA molecules of tospoviruses. These features of the viral genome are identical to those of tospoviruses; therefore, this virus is considered to belong to the genus Tospovirus. Its N protein comprised 279 amino acids and had a molecular mass of 31.0 kDa. Comparisons of its amino acid sequence with those of known tospoviruses revealed less than 60% identity. This melon virus is concluded to be a distinct species in the genus Tospovirus, and the name Melon yellow spot virus is proposed.

Kato K; Handa K; Kameya-Iwaki M

2000-04-01

316

Melon yellow spot virus: A Distinct Species of the Genus Tospovirus Isolated from Melon.  

Science.gov (United States)

ABSTRACT A tospovirus-like virus recovered from netted melon was transmitted by Thrips palmi in a persistent manner but had different cytopathological features from tospoviruses previously reported. Viral nucleocapsid (N) was purified with two protective reagents, 2-mercaptoethanol and L-ascorbic acid, and RNA extracted from the viral nucleocapsid was used for genomic analysis. The virus had a genome consisting of three single-stranded RNA molecules. The open reading frame on the viral complementary strand, located at the 3' end of the viral S RNA, encoded the N protein. The 3' terminus of this RNA also contained an eight-nucleotide sequence similar to the conserved sequence at the 3' end of genomic RNA molecules of tospoviruses. These features of the viral genome are identical to those of tospoviruses; therefore, this virus is considered to belong to the genus Tospovirus. Its N protein comprised 279 amino acids and had a molecular mass of 31.0 kDa. Comparisons of its amino acid sequence with those of known tospoviruses revealed less than 60% identity. This melon virus is concluded to be a distinct species in the genus Tospovirus, and the name Melon yellow spot virus is proposed. PMID:18944594

Kato, K; Handa, K; Kameya-Iwaki, M

2000-04-01

317

Vector-assisted seed transmission of melon necrotic spot virus in melon.  

UK PubMed Central (United Kingdom)

Seed lots prepared from fruits collected from systemically infected melon plants were tested for seedborne melon necrotic spot carmovirus (MNSV) The hypothesis of "vector-assisted" seed transmission (VAST) was tested in teals with rigorous control of the vector fungus, Olpidium bornovanus. The vector also formed the basis for the most sensitive assays of seedborne virus, enzyme-linked immunosorbent assay (ELISA) was intermediate, and infectivity was the least sensitive assay. Although extracts of seeds not treated with acid reacted in ELISA, the assay did not discriminate between viral antigens and infectious virus. The superficial viral antigens and virions were removed by acid treatment of seeds, but infectious virus remained within the seed coat at levels not usually detectable by ELISA. Seedborne virus rarely infected seedlings unless the vector was present, confirming VAST as a novel means of seed transmission that is possible because of the in vitro method of virus acquisition by O. bornovanus. The incidence of internally borne MNSV ranged from 0.1 to 5.3% in the vector-based assays. In one seed lot MNSV was primarily on the surface of seeds, with an incidence of >50% on nontreated seeds and 0% on acid-treated seeds.

Campbell RN; Wipf-Scheibel C; Lecoq H

1996-12-01

318

Association of Olpidium bornovanus and Melon necrotic spot virus with Vine Decline of Melon in Guatemala  

UK PubMed Central (United Kingdom)

Thirty-one soil samples from 14 different fields of Guatemala melon with vine decline symptoms were analyzed for the presence of organisms associated with the disease. With a soil-dilution plating method, only Macrophomina phaseolina was detected in five samples. With a melon bait plant technique, Olpidium bornovanus, often together with Melon necrotic spot virus (MNSV), was found in nearly all the samples, corresponding with all the fields studied. Other pathogens that were detected less frequently included Pythium aphanidermatum, Monosporascus cannonballus, and Rhizoctonia solani. Consequently, O. bornovanus and MNSV were uniquely associated with disease occurrence and thus are the most probable cause of melon vine decline in the fields studied.

Cara Mde; Lopez V; Cordoba MC; Santos M; Jorda C; Tello JC

2008-05-01

319

Mice with mutations of Dock7 have generalized hypopigmentation and white-spotting but show normal neurological function.  

UK PubMed Central (United Kingdom)

The classical recessive coat color mutation misty (m) arose spontaneously on the DBA/J background and causes generalized hypopigmentation and localized white-spotting in mice, with a lack of pigment on the belly, tail tip, and paws. Here we describe moonlight (mnlt), a second hypopigmentation and white-spotting mutation identified on the C57BL/6J background, which yields a phenotypic copy of m/m coat color traits. We demonstrate that the 2 mutations are allelic. m/m and mnlt/mnlt phenotypes both result from mutations that truncate the dedicator of cytokinesis 7 protein (DOCK7), a widely expressed Rho family guanine nucleotide exchange factor. Although Dock7 is transcribed at high levels in the developing brain and has been implicated in both axon development and myelination by in vitro studies, we find no requirement for DOCK7 in neurobehavioral function in vivo. However, DOCK7 has non-redundant role(s) related to the distribution and function of dermal and follicular melanocytes.

Blasius AL; Brandl K; Crozat K; Xia Y; Khovananth K; Krebs P; Smart NG; Zampolli A; Ruggeri ZM; Beutler BA

2009-02-01

320

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

Science.gov (United States)

Tomato spotted wilt virus (TSWV) virions consist of a nucleocapsid core surrounded by a membrane containing glycoproteins Gn and Gc. To unravel the protein interactions involved in the membrane acquisition of RNPs, TSWV nucleocapsid protein (N), Gn and Gc were expressed and analyzed in BHK21 cells. Upon coexpression of Gn, Gc and N, a partial colocalization of N with both glycoproteins was observed in the Golgi region. In contrast, upon coexpression of Gc and N in the absence of Gn, both proteins colocalized to a distinct non-Golgi perinuclear region. Using FLIM and FRET, interaction was demonstrated between N and Gc, but not between N and Gn, and was only observed in the region where both proteins accumulated. The genuine character of N-Gc interaction was confirmed by its presence in purified virus and RNP preparations. The results are discussed in view of TSWV particle assembly taking place at the Golgi complex. PMID:16963098

Snippe, Marjolein; Willem Borst, Jan; Goldbach, Rob; Kormelink, Richard

2006-09-11

 
 
 
 
321

Tomato spotted wilt virus Infection Improves Host Suitability for Its Vector Frankliniella occidentalis.  

UK PubMed Central (United Kingdom)

ABSTRACT The effect of Tomato spotted wilt virus (TSWV) infection on plant attractiveness for the western flower thrips (Frankliniella occidentalis) was studied. Significantly more thrips were recovered on infected than were recovered on noninfected pepper (Capsicum annuum) plants in different preference tests. In addition, more offspring were produced on the virus-infected pepper plants, and this effect also was found for TSWV-infected Datura stramonium. Thrips behavior was minimally influenced by TSWV-infection of host plants with only a slight preference for feeding on infected plants. Offspring development was positively affected since larvae hatched earlier from eggs and subsequently pupated faster on TSWV-infected plants. These results show a mutualistic relationship between F. occidentalis and TSWV.

Maris PC; Joosten NN; Goldbach RW; Peters D

2004-07-01

322

Tomato spotted wilt virus Infection Improves Host Suitability for Its Vector Frankliniella occidentalis.  

Science.gov (United States)

ABSTRACT The effect of Tomato spotted wilt virus (TSWV) infection on plant attractiveness for the western flower thrips (Frankliniella occidentalis) was studied. Significantly more thrips were recovered on infected than were recovered on noninfected pepper (Capsicum annuum) plants in different preference tests. In addition, more offspring were produced on the virus-infected pepper plants, and this effect also was found for TSWV-infected Datura stramonium. Thrips behavior was minimally influenced by TSWV-infection of host plants with only a slight preference for feeding on infected plants. Offspring development was positively affected since larvae hatched earlier from eggs and subsequently pupated faster on TSWV-infected plants. These results show a mutualistic relationship between F. occidentalis and TSWV. PMID:18943902

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

2004-07-01

323

Detection of tomato spotted wilt virus using monoclonal antibodies and riboprobes.  

UK PubMed Central (United Kingdom)

The immunoreactivity of a panel of monoclonal antibodies raised to tomato spotted wilt virus (TSWV) was examined in enzyme-linked immunosorbent assays (ELISA) and dot immunobinding assays (DIBA) procedures. MAbs 6.12.15 and 2.9 were specific for the nucleocapsid protein of TSWV. The sensitivity of the two immunoassays was compared with that of a dot-blot hybridization technique using riboprobes (RNA transcripts) to TSWV M RNA. Using deproteinized plant extracts or purified virus preparations, as little as 1 pg RNA could be detected. Although an ELISA using MAb 6.12.15, a DIBA procedure using MAb 3.22.6 and the dot-blot hybridization, detected several TSWV isolates in different host species equally well, the ELISA was most precise and most suitable for routine diagnosis in the field.

Huguenot C; van den Dobbelsteen G; de Haan P; Wagemakers CA; Drost GA; Osterhaus AD; Peters D

1990-01-01

324

Distribution of apple stem grooving virus and apple chlorotic leaf spot virus in infected in vitro pear shoots  

UK PubMed Central (United Kingdom)

The distribution patterns of Apple stem grooving virus (ASGV) and Apple chlorotic leaf spot virus (ACLSV) in in vitro-cultured pear plants were investigated using in situ tissue-printing hybridization (TPH) and tissue blotting immunoassay (TBIA) to detect viral RNAs and coating proteins. Both ASGV and ACLSV showed high concentrations in the tip of the pear shoots and lower concentrations in the middle stem. The highest viral RNA titers were found in the phloem parenchyma of vascular bundles. Monitoring of viral RNA concentrations was conducted on infected in vitro-cultured pear plants during thermotherapy using TPH combined with X-ray film exposure in serial cross sections. No viral RNA of ACLSV or ASGV was detected in less than 2 mm and 0.5 mm long tips, respectively. The heat treatment was less effective to reduce virus titers in the bottom shoot. The obtained results would assist in the selection of tips with proper sizes from pear shoots pre- and post-thermotherapy for the production of virus-free pear plants by meristem culture.

Wang LP; Hong N; Wang GP; Xu WX; Michelutti R; Wang AM

2010-12-01

325

Induction of necrosis via mitochondrial targeting of Melon necrotic spot virus replication protein p29 by its second transmembrane domain  

International Nuclear Information System (INIS)

The virulence factor of Melon necrotic spot virus (MNSV), a virus that induces systemic necrotic spot disease on melon plants, was investigated. When the replication protein p29 was expressed in N. benthamiana using a Cucumber mosaic virus vector, necrotic spots appeared on the leaf tissue. Transmission electron microscopy revealed abnormal mitochondrial aggregation in these tissues. Fractionation of tissues expressing p29 and confocal imaging using GFP-tagged p29 revealed that p29 associated with the mitochondrial membrane as an integral membrane protein. Expression analysis of p29 deletion fragments and prediction of hydrophobic transmembrane domains (TMDs) in p29 showed that deletion of the second putative TMD from p29 led to deficiencies in both the mitochondrial localization and virulence of p29. Taken together, these results indicated that MNSV p29 interacts with the mitochondrial membrane and that p29 may be a virulence factor causing the observed necrosis.

2009-08-01

326

Soybean chlorotic spot virus, a novel begomovirus infecting soybean in Brazil.  

UK PubMed Central (United Kingdom)

A novel soybean-infecting begomovirus from Brazil was identified in Jaíba, in the state of Minas Gerais, and molecularly characterized. By using rolling-circle amplification-based cloning of viral DNAs, three DNA-A variants and a cognate DNA-B were isolated from infected samples. The DNA variants share more than 98 % sequence identity but have less than 89 % identity to other reported begomovirus, the limit for demarcation of new species. In a phylogenetic analysis, both DNA-A and DNA-B clustered with other Brazilian begomoviruses. Infectious cloned DNA-A and DNA-B components induced distinct symptoms in Solanaceae and Fabaceae species by biolistic inoculation. In soybean, the virus induced mild symptoms, i.e., chlorotic spots on the leaves, from which the name soybean chlorotic spot virus (SoCSV) was proposed. The most severe symptoms were displayed by common beans, which exhibited leaf distortion, blistering, interveinal chlorosis, mosaic and golden mosaic. The possibility that SoCSV may become a threat to bean production in Brazil is discussed.

Coco D; Calil IP; Brustolini OJ; Santos AA; Inoue-Nagata AK; Fontes EP

2013-02-01

327

Coincident ruddy turnstone migration and horseshoe crab spawning creates an ecological 'hot spot' for influenza viruses.  

UK PubMed Central (United Kingdom)

Since 1985, avian influenza virus surveillance has been conducted annually from mid-May to early June in charadriiform species from the families Scolopacidae and Laridae (shorebirds and gulls) at Delaware Bay in the northeast United States. The mass migrations of shorebirds, gulls and horseshoe crabs (Limulus polyphemus) coincide at that time, and large numbers of migrating birds pause at Delaware Bay to feed on horseshoe crab eggs deposited at the high-tide line. Influenza viruses are consistently isolated from charadriiform birds at Delaware Bay, at an overall rate approximately 17 times the combined rate of isolation at all other surveillance sites worldwide (490 isolates/9474 samples, 5.2% versus 49 isolates per 15,848 samples, 0.3%, respectively; Proportion test, p < 0.0001). The likelihood of isolating influenza viruses at Delaware Bay is dependent on the presence of ruddy turnstone (Arenaria interpres) at the sampling site (G-test of independence, p < 0.001). The convergence of host factors and environmental factors results in a unique ecological 'hot spot' for influenza viruses in Charadriiformes.

Krauss S; Stallknecht DE; Negovetich NJ; Niles LJ; Webby RJ; Webster RG

2010-11-01

328

Elastomeric-ligated vs self-ligating appliances: a pilot study examining microbial colonization and white spot lesion formation after 1 year of orthodontic treatment.  

UK PubMed Central (United Kingdom)

AIM: To (1) evaluate the use of adenosine triphosphate (ATP)-driven bioluminescence for quantification of total plaque bacteria in orthodontic patients, (2) compare plaque bacteria amounts at the bracket-tooth interface with use of elastomeric-ligated and self-ligating brackets after 1 year of orthodontic treatment, and (3) analyze formation of white spot lesions by photographic evaluation and laser-light fluorescence (DIAGNOdent). METHODS: Thirteen subjects had fixed orthodontic appliances placed where lateral incisors were bonded with either elastomeric-ligated or self-ligating brackets. Plaque bacteria were collected from incisor surfaces after 1 year and quantified using plating methods and ATP-driven bioluminescence. White spot lesions were evaluated by photographic and DIAGNOdent determinations. A 2 x 2 x 2 mixed-design ANOVA was conducted to determine differences in plaque retention between elastomeric-ligated and self-ligating brackets. RESULTS: ATP-driven bioluminescence values correlated to numbers of total plaque bacteria (r = 0.80). However, unlike findings published in the original pilot study, which described increased plaque retention with elastomeric-ligated brackets at 5 weeks postbonding, there were no significant differences in bacterial numbers or ATP-driven bioluminescence values surrounding the elastomeric-ligated vs self-ligating brackets after 1 year of orthodontic treatment. Based on photographic and DIAGNOdent determinations, white spot lesions were found relatively equally on teeth bonded with either bracket type. DIAGNOdent measurements were found to have moderate sensitivity (0.71) and good specificity (0.88) when compared to white spot lesions determined using photographic evaluation. CONCLUSION: ATP-driven bioluminescence can be used as an accurate assessment of total plaque bacteria in orthodontic patients. After 1 year of orthodontic treatment for patients in this pilot study, there appeared to be no differences in retention of plaque bacteria or white spot lesions comparing the bracket types. The use of DIAGNOdent has some limitations, but may prove to be useful to monitor white spot lesions longitudinally.

Buck T; Pellegrini P; Sauerwein R; Leo MC; Covell DA Jr; Maier T; Machida CA

2011-01-01

329

A new cryptic virus belonging to the family Partitiviridae was found in watermelon co-infected with Melon necrotic spot virus.  

Science.gov (United States)

A novel virus was detected in watermelon plants (Citrullus lanatus Thunb.) infected with Melon necrotic spot virus (MNSV) using SOLiD next-generation sequence analysis. In addition to the expected MSNV genome, two double-stranded RNA (dsRNA) segments of 1,312 and 1,118 bp were also identified and sequenced from the purified virus preparations. These two dsRNA segments encode two putative partitivirus-related proteins, an RNA-dependent RNA polymerase (RdRP) and a capsid protein, which were sequenced. Genomic-sequence analysis and analysis of phylogenetic relationships indicate that these two dsRNAs together make up the genome of a novel Partitivirus. This virus was found to be closely related to the Pepper cryptic virus 1 and Raphanus sativus cryptic virus. It is suggested that this novel virus putatively named Citrullus lanatus cryptic virus be considered as a new member of the family Partitiviridae. PMID:23775759

Sela, Noa; Lachman, Oded; Reingold, Victoria; Dombrovsky, Aviv

2013-06-18

330

A new cryptic virus belonging to the family Partitiviridae was found in watermelon co-infected with Melon necrotic spot virus.  

UK PubMed Central (United Kingdom)

A novel virus was detected in watermelon plants (Citrullus lanatus Thunb.) infected with Melon necrotic spot virus (MNSV) using SOLiD next-generation sequence analysis. In addition to the expected MSNV genome, two double-stranded RNA (dsRNA) segments of 1,312 and 1,118 bp were also identified and sequenced from the purified virus preparations. These two dsRNA segments encode two putative partitivirus-related proteins, an RNA-dependent RNA polymerase (RdRP) and a capsid protein, which were sequenced. Genomic-sequence analysis and analysis of phylogenetic relationships indicate that these two dsRNAs together make up the genome of a novel Partitivirus. This virus was found to be closely related to the Pepper cryptic virus 1 and Raphanus sativus cryptic virus. It is suggested that this novel virus putatively named Citrullus lanatus cryptic virus be considered as a new member of the family Partitiviridae.

Sela N; Lachman O; Reingold V; Dombrovsky A

2013-06-01

331

A new cryptic virus belonging to the family Partitiviridae was found in watermelon co-infected with Melon necrotic spot virus.  

UK PubMed Central (United Kingdom)

A novel virus was detected in watermelon plants (Citrullus lanatus Thunb.) infected with Melon necrotic spot virus (MNSV) using SOLiD next-generation sequence analysis. In addition to the expected MSNV genome, two double-stranded RNA (dsRNA) segments of 1,312 and 1,118 bp were also identified and sequenced from the purified virus preparations. These two dsRNA segments encode two putative partitivirus-related proteins, an RNA-dependent RNA polymerase (RdRP) and a capsid protein, which were sequenced. Genomic-sequence analysis and analysis of phylogenetic relationships indicate that these two dsRNAs together make up the genome of a novel Partitivirus. This virus was found to be closely related to the Pepper cryptic virus 1 and Raphanus sativus cryptic virus. It is suggested that this novel virus putatively named Citrullus lanatus cryptic virus be considered as a new member of the family Partitiviridae.

Sela N; Lachman O; Reingold V; Dombrovsky A

2013-10-01

332

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

UK PubMed Central (United Kingdom)

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

García-Marcos A; Pacheco R; Manzano A; Aguilar E; Tenllado F

2013-05-01

333

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

Science.gov (United States)

One of the most severe symptoms caused by compatible plant-virus interactions is systemic necrosis, which shares common attributes with the hypersensitive response to incompatible pathogens. Although several studies have identified viral symptom determinants responsible for systemic necrosis, mechanistic models of how they contribute to necrosis in infected plants remain scarce. Here, we examined the involvement of different branches of the oxylipin biosynthesis pathway in the systemic necrosis response caused either by the synergistic interaction of Potato virus X with Potato virus Y (PVX-PVY) or by Tomato spotted wilt virus (TSWV) in Nicotiana benthamiana. Silencing either 9-lipoxygenase (LOX), 13-LOX, or ?-dioxygenase-1 (?-DOX-1) attenuated the programmed cell death (PCD)-associated symptoms caused by infection with either PVX-PVY or TSWV. In contrast, silencing of the jasmonic acid perception gene, COI1 (Coronatine insensitive 1), expedited cell death during infection with compatible viruses. This correlated with an enhanced expression of oxylipin biosynthesis genes and dioxygenase activity in PVX-PVY-infected plants. Moreover, the Arabidopsis thaliana double lox1 ?-dox-1 mutant became less susceptible to TSWV infection. We conclude that oxylipin metabolism is a critical component that positively regulates the process of PCD during compatible plant-virus interactions but does not play a role in restraining virus accumulation in planta. PMID:23487466

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

2013-03-13

334

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

Digital Repository Infrastructure Vision for European Research (DRIVER)

Background: Nowadays, dried blood spots (DBS) are primarily used to obtain diagnostic access to risk collectives such as intravenous drug users, who are prone to infections with hepatitis B virus (HBV), hepatitis C virus (HCV), and human immunodeficiency virus (HIV). Before DBS analyses ca...

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

335

Synergistic Interaction Between Tomato chlorosis virus and Tomato spotted wilt virus Results in Breakdown of Resistance in Tomato.  

Science.gov (United States)

ABSTRACT Multiple viral infections frequently are found in single plants of cultivated and wild hosts in nature, with unpredictable pathological consequences. Synergistic reactions were observed in mixed infections in tomato plants doubly infected with the positive-sense and phloem-limited single-stranded RNA (ssRNA) crinivirus Tomato chlorosis virus (ToCV) and the negative-sense ssRNA tospovirus Tomato spotted wilt virus (TSWV). Synergism in a tomato cultivar susceptible to both viruses resulted in a rapid death of plants. A pronounced enhancement of ToCV accumulation mediated by TSWV co-infection was observed with no evident egress of ToCV from phloem tissues. No consistent alteration of TSWV accumulation was detected. More remarkable was the synergism observed in tomato cultivars which carry the Sw-5 resistance gene, which are resistant to TSWV. Pre-infection with ToCV resulted in susceptibility to TSWV, whereas co-inoculations did not. This suggested that a threshold level or a time lapse is needed for ToCV to interfere or downregulate the defense response in the TSWV-resistant plants. PMID:18943964

García-Cano, Elena; Resende, Renato O; Fernández-Muñoz, Rafael; Moriones, Enrique

2006-11-01

336

First Report of Impatiens necrotic spot virus on Spiderlily in China  

UK PubMed Central (United Kingdom)

Impatiens necrotic spot virus (INSV) (genus Tospovirus; family Bunyaviridae) is a devastating disease in the production of ornamental flowers (1). From 2007 to 2009, a survey of 10 major parks and recreation areas in Kunming, the capital of Yunnan Province, China, indicated that approximately 60 to 70% of Spiderlily (Hymenocallis littoralis Salisb.) plants from eight parks had symptoms of concentric ring spots and necrotic spots. Symptomatic plants were tested for INSV and Tomato spotted wilt virus (TSWV) with an immunostrip (Agdia Inc. Elkhart, IN). Results indicated that only the samples designated HDL were positive for INSV and all other samples were negative for both INSV and TSWV. Mechanically inoculated Emilia sonchifolia, Nicotiana glutinosa, Impatiens balsamina, and N. rustica showed chlorotic lesions, concentric rings, and severe necrosis, symptoms typical for INSV in these hosts. Electron microscope inspection found tospovirus-like spheroidal, enveloped particles that were 90 nm in diameter. Primer 5 software (Premier, Canada) was used to design 14 primers from GenBank Accession No. NC_003625 to amplify the L RNA, nine from NC_003616 to amplify the M RNA, and six from NC_003624 to amplify the S RNA. With total RNA extracted from infected plant tissue as templates in reverse transcription (RT)-PCR, these primers generated 29 target fragments of 250 to 900 bp. These fragments were cloned with the vector pMD19 simple-T vector (Takara Bio Inc., Dalian, China) and sequenced. The sequences of the clones were aligned with the software DNAman (version 2.5; Lynnon Biosoft, Quebec, Canada), showing that RNAs L, M, and S are 8,776 bp (GenBank Accession No. GU112505), 4,948 bp (GenBank Accession No. GU112503), and 2,875 bp (GenBank Accession No. GU112504), respectively. BLAST analysis of these Spiderlily INSV sequences against the NCBI sequence database indicated that the RdRp protein (L RNA) was 99.6% identical with the RdRp protein from an Italian isolate (No. DQ425094), the Nsm protein (M RNA) has 99.0% identity with the Nsm protein from an isolate from Italy (No. DQ425095) and one from the United States (No. NC_003616), the G1G2 polyprotein (M RNA) has 99.9% identity with the analogous protein from an Italian isolate (No. DQ425095), the N protein (S RNA) has 99.6% identity with the N protein from an Italian isolate (No. DQ425096), and the NSs protein (S RNA) has 98.7% identity with the NSs protein from an isolate from Japan (No. AB109100). To our knowledge, this is the first report of INSV on Spiderlily in China.

Liu YT; Zheng YX; Li YZ; Li ZY

2010-04-01

337

Lack of reliable evidence of the effectiveness of remineralising agents for the treatment of post orthodontic white spot lesions.  

UK PubMed Central (United Kingdom)

Data sourcesPubMed, Medline, Web of Science and the Cochrane Library databases were searched.Study selectionSelection was conducted independently by two reviewers, only randomised controlled trials were included. Study quality was assessed using the Cochrane risk of bias approach.Data extraction and synthesisData abstraction was carried out independently by two reviewers and qualitative summary presented, because of the heterogeneity of the interventions and outcome measures.ResultsSeven studies were included, all except one (of medium risk) were considered to be at high risk of bias. Three studies evaluated fluoride preparations (50-ppm sodium fluoride mouth rinse, 5% sodium fluoride varnish and 0.5% sodium fluoride chewing sticks). Four compared remineralising agents containing casein phosphopeptide amorphous calcium phosphate or casein phosphopeptide amorphous calcium fluoride phosphate.ConclusionsBased on the literature, there is a lack of reliable evidence to support the effectiveness of remineralising agents for the treatment of post orthodontic white spot lesions.

Kalha AS

2013-09-01

338

Bioaccumulation of organochlorines in relation to the life history in the white-spotted charr Salvelinus leucomaenis.  

UK PubMed Central (United Kingdom)

The bioaccumulation of organochlorines (OCs) in the muscle tissue of sea-run (anadromous) and freshwater-resident (fluvial) white-spotted charr (Salvelinus leucomaenis) was determined to assess the ecological risk related to intraspecies variations in diadromous fish life history as they migrate between sea and freshwater. Generally, there were significant correlations between the accumulation of OCs such as DDTs, HCB, HCHs and CHLs. In addition, various biological characteristics, such as total length (TL), body weight (BW) and age, and number of downstream migration (NDM) were correlated. A positive correlation occurred between the lipid content and the OC concentrations. Close linear relationships were found between TL, BW and NDM and the lipid content. Although they are both the same species, the OCs concentrations in the anadromous fish were significantly higher than those in the fluvial individuals. These results suggest that anadromous S. leucomaenis have a higher ecological risk for OCs exposure than the fluvial fish.

Arai T

2013-02-01

339

Infiltration, a new therapy for masking enamel white spots: a 19-month follow-up case series.  

Science.gov (United States)

Enamel white spot lesions are frequent and can impact patients' quality of life. The most conservative treatment in such cases is microabrasion, a technique that presents some drawbacks. The proposed strategy is not based on the elimination of dysplastic enamel, but on masking the lesion by infiltrating the porous subsurface enamel with a hydrophobic resin that has a refraction index closer to that of sound enamel, after permeating the non-porous surface enamel through hydrochloric acid erosion. Erosion-infiltration approaches have been proposed to treat initial caries, but this report suggests extending it to two novel indications: fluorosis and traumatic hypo-mineralization lesions. Four cases were treated by erosion infiltration following the original protocol. They were followed up clinically at several intervals during a period of 19 months of clinical service. The clinical results, although not perfect, satisfied the patients entirely. Erosion infiltration could be a promising alternative for minimally invasive treatment in similar situations. PMID:23712339

Tirlet, Gil; Chabouis, Hélène Fron; Attal, Jean-Pierre

2013-01-01

340

Infiltration, a new therapy for masking enamel white spots: a 19-month follow-up case series.  

UK PubMed Central (United Kingdom)

Enamel white spot lesions are frequent and can impact patients' quality of life. The most conservative treatment in such cases is microabrasion, a technique that presents some drawbacks. The proposed strategy is not based on the elimination of dysplastic enamel, but on masking the lesion by infiltrating the porous subsurface enamel with a hydrophobic resin that has a refraction index closer to that of sound enamel, after permeating the non-porous surface enamel through hydrochloric acid erosion. Erosion-infiltration approaches have been proposed to treat initial caries, but this report suggests extending it to two novel indications: fluorosis and traumatic hypo-mineralization lesions. Four cases were treated by erosion infiltration following the original protocol. They were followed up clinically at several intervals during a period of 19 months of clinical service. The clinical results, although not perfect, satisfied the patients entirely. Erosion infiltration could be a promising alternative for minimally invasive treatment in similar situations.

Tirlet G; Chabouis HF; Attal JP

2013-01-01

 
 
 
 
341

Development of a real-time fluorescent quantitative PCR assay for detection of Impatiens necrotic spot virus.  

Science.gov (United States)

Impatiens necrotic spot virus (INSV) is an important plant virus that can cause severe disease in various ornamental and agricultural crops. Several species of thrips transmit INSV, of which the western flower thrip (Frankliniella occidentalis) is the most important. In this study, primers and TaqMan probes based on INSV non-structural protein gene sequences were designed, and a technique was developed for detecting INSV using fluorescent quantitative RT-PCR. The reproducibility, specificity and sensitivity for the RT-PCR were evaluated; and the RT-PCR method was developed to detect INSV in the host plants and western flower thrips. A standard curve constructed by a series of diluted plasmid DNA gave a good linear relationship between Ct value and concentration of plasmid DNA, a low coefficient of variation and good reproducibility. The detection method not only measured quantitatively the concentration of INSV in plant hosts and western flower thrips, but also measured accurately low concentrations of the virus. The measurable concentration fell to as low as 10? copies/?l, while RT-PCR could detect only 10² copies/?l. The method had high specificity and could distinguish INSV from Tomato spotted wilt virus (TSWV) and Tomato zonate spot virus (TZSV), both from the same genus of viruses. This is the first report of the same method being used to detect INSV in both plant hosts and western flower thrips, and should be helpful in studies of INSV epidemiology. PMID:23466630

Chen, Xuejiao; Xu, Xiaogang; Li, Yongzhong; Liu, Yating

2013-03-04

342

Development of a real-time fluorescent quantitative PCR assay for detection of Impatiens necrotic spot virus.  

UK PubMed Central (United Kingdom)

Impatiens necrotic spot virus (INSV) is an important plant virus that can cause severe disease in various ornamental and agricultural crops. Several species of thrips transmit INSV, of which the western flower thrip (Frankliniella occidentalis) is the most important. In this study, primers and TaqMan probes based on INSV non-structural protein gene sequences were designed, and a technique was developed for detecting INSV using fluorescent quantitative RT-PCR. The reproducibility, specificity and sensitivity for the RT-PCR were evaluated; and the RT-PCR method was developed to detect INSV in the host plants and western flower thrips. A standard curve constructed by a series of diluted plasmid DNA gave a good linear relationship between Ct value and concentration of plasmid DNA, a low coefficient of variation and good reproducibility. The detection method not only measured quantitatively the concentration of INSV in plant hosts and western flower thrips, but also measured accurately low concentrations of the virus. The measurable concentration fell to as low as 10? copies/?l, while RT-PCR could detect only 10² copies/?l. The method had high specificity and could distinguish INSV from Tomato spotted wilt virus (TSWV) and Tomato zonate spot virus (TZSV), both from the same genus of viruses. This is the first report of the same method being used to detect INSV in both plant hosts and western flower thrips, and should be helpful in studies of INSV epidemiology.

Chen X; Xu X; Li Y; Liu Y

2013-05-01

343

Monoclonal Antibodies against the Recombinant Nucleocapsid Protein of Tomato spotted wilt virus and its Application in Virus Detection  

UK PubMed Central (United Kingdom)

Tomato spotted wilt virus (TSWV) is the type member of the tospovirus genus and causes significant losses in a wide range of economically important ornamental and vegetable crops worldwide. The nucleocapsid gene, located on the ambisense S RNA segment of TSWV was expressed in Escherichia coli using pET-32a as vector and correct expression of recombinant protein was confirmed by Western blot using an anti-TSWV monoclonal antibody (MAb). The recombinant protein was purified using Ni-NTA agarose and the purified protein was used for the production of MAbs. Three murine MAbs against the recombinant nucleocapsid protein were produced. Triple antibody sandwich enzyme-linked immunosorbent assay and immunocapture RT-PCR methods were then established for reliable and efficient detection of TSWV using the produced MAbs.

Wu Jianxiang; Yu Cui; Yang Cuiyun; Zhou Xueping

2009-06-01

344

TOMATO SPOTTED WILT VIRUS in PEANUT TISSUE TYPES AND PHYSIOLOGICAL EFFECTS RELATED TO disease incidence and SEVERITY  

Science.gov (United States)

Much has been speculated about whether certain physiological characteristics in peanut varieties enable more resistant varieties to withstand tomato spotted wilt virus (TSWV) infection better than others. In order to address this question, three peanut varieties, Georgia Green, NC-V11, and ANorden,...

345

A novel polymerase chain reaction method for detection of human immunodeficiency virus in dried blood spots on filter paper.  

Digital Repository Infrastructure Vision for European Research (DRIVER)

A method for detection of proviral human immunodeficiency virus DNA in dried blood spots on filter paper by direct polymerase chain reaction (PCR) has been developed. To develop the method, a standard system was used which was prepared from cells each containing a single integrated provirus and titr...

Yourno, J; Conroy, J

346

SEASONAL DISPERSAL PATTERNS OF FRANKLINIELLA FUSCA (THYSANOPTERA: THRIPIDAE) AND TOMATO SPOTTED WILT VIRUS OCCURRENCE IN CENTRAL AND EASTERN NORTH CAROLINA  

Science.gov (United States)

The seasonal abundance and temporal pattern of Frankliniella fusca Hinds dispersal were monitored from 1996 to 2000 at 12 locations in central and eastern North Carolina. The predominant vector of tomato spotted wilt virus (TSWV) vector species captured across all locations was F. fusca (98%). The...

347

Application of Dried Blood Spot Specimens for Serologic Subtyping of Human Immunodeficiency Virus Type 1 in Thailand  

Digital Repository Infrastructure Vision for European Research (DRIVER)

Dried blood spot (DBS) specimens were assessed as an alternative to plasma for human immunodeficiency virus type 1 (HIV-1) serotyping by V3 loop peptide enzyme immunoassay. Nested PCR capable of distinguishing HIV-1 subtypes B and E was used as the reference standard. Ninety-two percent of DBS sampl...

Chanbancherd, P.; Brown, A. E.; Trichavaroj, R.; Tienamporn, P.; Puthakird, P.; Limpairojn, N.; VanCott, T. C.; de Souza, M. S.

348

Use of dried blood spot specimens in the detection of human immunodeficiency virus type 1 by the polymerase chain reaction.  

Digital Repository Infrastructure Vision for European Research (DRIVER)

Dried blood spots (DBSs) constitute a potentially valuable source of material for human immunodeficiency virus (HIV) serologic and molecular testing. To facilitate molecular testing, we have adapted the polymerase chain reaction (PCR) to the detection of HIV proviral DNA in DBS samples. The method i...

Cassol, S; Salas, T; Arella, M; Neumann, P; Schechter, M T; O'Shaughnessy, M

349

Detection of Human Immunodeficiency Virus Type 1 DNA in Dried Blood Spots by a Duplex Real-Time PCR Assay  

Digital Repository Infrastructure Vision for European Research (DRIVER)

A dried blood spot (DBS) is a well-accepted means for the collection, transport, and storage of blood samples for various epidemiologic, serologic, and molecular assays for human immunodeficiency virus (HIV) studies. It is particularly important for mother-to-infant-transmission studies of affected ...

Luo, Wei; Yang, Hua; Rathbun, Kimberly; Pau, Chou-Pong; Ou, Chin-Yih

350

Detection of Antibodies to Hepatitis C Virus in Dried Blood Spot Samples from Mothers and Their Offspring in Lahore, Pakistan  

Digital Repository Infrastructure Vision for European Research (DRIVER)

Dried blood spot samples from mothers and their offspring attending the obstetric and pediatric departments of two hospitals in Lahore, Pakistan, were tested for antibodies to hepatitis C virus (HCV). The seroprevalence of HCV in the women was 6.7% (95% confidence interval [CI], 4.3 to 9.1), and tha...

Parker, S. P.; Khan, H. I.; Cubitt, W. D.

351

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

Science.gov (United States)

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...

352

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

UK PubMed Central (United Kingdom)

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.

López C; Aramburu J; Galipienso L; Soler S; Nuez F; Rubio L

2011-01-01

353

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

Science.gov (United States)

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. PMID:20881087

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

2010-09-29

354

A Pathotype of Pepper mild mottle virus Causing Necrotic Spot Symptoms in Paprika Fruit  

Directory of Open Access Journals (Sweden)

Full Text Available Black necrotic spots were observed from the fruits of paprika that were cultivating in a vinylhouse. Thecasual agents of the symptom were identified as several isolates of Pepper mild mottle virus (PMMoV) byresponses of indicator plants, electron microscopy, and RT-PCR analysis. Symptoms of the viral disease weremild mottle in the young leaves, necrotic spots on the fruits and the fruit apex of paprika, but the symptomswere not shown on the mature leaves. All of the PMMoV isolates were determined as P1.2.3 pathotypes from thebiological responses on the chilli pepper lines used for discrimination of tobamovirus pathotypes.Pathogenicity of the PMMoV isolates was also confirmed using mechanical inoculation method to paprikaseedlings. The coat protein (CP) genes of the PMMoV isolates were compared at the nucleotide and aminoacid levels with the previously published PMMoV isolate. The isolates share 96 to 99% CP nucleotide identityamong the isolates. The CP of P1.2-pathotype PMMoV-P2 presented Met at position 139, But the CPs of P1.2.3-pathotype PMMoVs from paprika showed Met to Asn substitution at the same position. This is the firstreport of identification of P1.2.3-pathotype PMMoV isolates from paprika in Korea.

Gug-Seoun Choi; Sung-Kook Choi; Jeom-Deog Cho; In-Sook Cho

2013-01-01

355

Tactics for management of thrips (Thysanoptera: Thripidae) and tomato spotted wilt virus in tomato.  

Science.gov (United States)

Four studies were conducted in Georgia during spring 1999, 2000, 2001, and 2002 to evaluate various management tactics for reducing thrips and thrips-vectored tomato spotted wilt virus (TSWV) in tomato and their interactions relative to fruit yield. Populations of thrips vectors of TSWV, Frankliniella occidentalis (Pergande) and Frankliniella fusca (Hinds), were determined using flower and sticky trap samples. The management practices evaluated were host plant resistance, insecticide treatments, and silver or metallic reflective mulch. Averaged over all tests, the TSWV-resistant tomato 'BHN444' on silver mulch treatment had the largest effect in terms of reducing thrips and spotted wilt and increasing marketable yield. Of the insecticide treatments tested, the imidacloprid soil treatment followed by early applications of a thrips-effective foliar insecticide treatment provided significant increase in yield over other treatments. Tomato yield was negatively correlated with the number of F. fusca and percentage of TSWV incidence. F. occidentalis per blossom was positively correlated with percentage of TSWV incidence, but not with yield. No significant interactions were observed between cultivar reflective mulch main plot treatments and insecticide subplot treatments; thus, treatment seemed to be additive in reducing the economic impact of thrips-vectored TSWV. Control tactics that manage thrips early in the growing season significantly increased tomato yield in years when the incidence of TSWV was high (>17%). PMID:15568355

Riley, D G; Pappu, H R

2004-10-01

356

Tactics for management of thrips (Thysanoptera: Thripidae) and tomato spotted wilt virus in tomato.  

UK PubMed Central (United Kingdom)

Four studies were conducted in Georgia during spring 1999, 2000, 2001, and 2002 to evaluate various management tactics for reducing thrips and thrips-vectored tomato spotted wilt virus (TSWV) in tomato and their interactions relative to fruit yield. Populations of thrips vectors of TSWV, Frankliniella occidentalis (Pergande) and Frankliniella fusca (Hinds), were determined using flower and sticky trap samples. The management practices evaluated were host plant resistance, insecticide treatments, and silver or metallic reflective mulch. Averaged over all tests, the TSWV-resistant tomato 'BHN444' on silver mulch treatment had the largest effect in terms of reducing thrips and spotted wilt and increasing marketable yield. Of the insecticide treatments tested, the imidacloprid soil treatment followed by early applications of a thrips-effective foliar insecticide treatment provided significant increase in yield over other treatments. Tomato yield was negatively correlated with the number of F. fusca and percentage of TSWV incidence. F. occidentalis per blossom was positively correlated with percentage of TSWV incidence, but not with yield. No significant interactions were observed between cultivar reflective mulch main plot treatments and insecticide subplot treatments; thus, treatment seemed to be additive in reducing the economic impact of thrips-vectored TSWV. Control tactics that manage thrips early in the growing season significantly increased tomato yield in years when the incidence of TSWV was high (>17%).

Riley DG; Pappu HR

2004-10-01

357

First report of onion (Allium cepa) naturally infected with Iris yellow spot virus in Peru.  

UK PubMed Central (United Kingdom)

Onions have become an important export crop for Peru during the last few years. The onions produced for export are primarily short-day onions and include Grano- or Granex-type sweet onions. The first of two growing seasons for onion in Peru occurs from February/March until September/October and the second occurs from September/October to December/January. Iris yellow spot virus (IYSV [family Bunyaviridae, genus Tospovirus]), primarily transmitted by onion thrips (Thrips tabaci), has been reported in many countries during recent years, including the United States (1,2). In South America, the virus was reported in Brazil during 1999 (3) and most recently in Chile during 2005 (4). During 2003, an investigation of necrotic lesions and dieback in onions grown near the towns of Supe and Ica, Peru led to the discovery of IYSV in this region. Of 25 samples of symptomatic plants collected from five different fields near Supe, 19 tested strongly positive and an additional three tested weakly positive for IYSV using double antibody sandwich-enzyme linked immunosorbent assay (DAS-ELISA) (Agdia Inc., Elkhart, IN). None of the samples tested positive for Tomato spotted wilt virus (TSWV). A number of onions with necrosis and dieback symptoms were also observed during 2004 and 2005. During September 2005, 25 plants with symptoms suspected to be caused by IYSV or TSWV in the Supe and Casma valleys were collected and screened for both viruses using DAS-ELISA. All plants screened were positive for IYSV. There was no serological indication of TSWV infection in these samples. The positive samples were blotted onto FTA cards (Whatman Inc., U.K.) to bind the viral RNA for preservation and processed according to the manufacturer's protocols. The presence of IYSV was verified by reverse transcription-polymerase chain reaction (RTPCR) using (5'-TCAGAAATCGAGAAACTT-3') and (5'-TAATTATATCTATCTTTCTTGG-3') as forward and reverse primers (1), respectively. The primers amplify the nucleocapsid (N) gene of IYSV, and the RT-PCR products from this reaction were analyzed with gel electrophoresis with an ethidium bromide stain in 0.8% agarose to verify the presence of this amplicon in the samples. Subsequent to the September 2005 sampling, 72 additional samples from regions in northern and southern Peru were analyzed in the same manner. The amplicons obtained were cloned, sequenced, and compared with known IYSV isolates for further verification. Onions have become a significant export crop for Peru, and more research is needed to determine the impact of IYSV on the Peruvian onion export crop. To our knowledge, this is the first report of IYSV in onion in Peru.

Mullis SW; Gitaitis RD; Nischwitz C; Csinos AS; Mallaupoma ZCR; Rojas EHI

2006-03-01

358

A predictive model for spotted wilt epidemics in peanut based on local weather conditions and the tomato spotted wilt virus risk index.  

Science.gov (United States)

Tomato spotted wilt virus (TSWV), a member of the genus Tospovirus (family Bunyaviridae), is an important plant virus that causes severe damage to peanut (Arachis hypogaea) in the southeastern United States. Disease severity has been extremely variable in individual fields in Georgia, due to several factors including variability in weather patterns. A TSWV risk index has been developed by the University of Georgia to aid peanut growers with the assessment and avoidance of high risk situations. This study was conducted to examine the relationship between weather parameters and spotted wilt severity in peanut, and to develop a predictive model that integrates localized weather information into the risk index. On-farm survey data collected during 1999, 2002, 2004, and 2005 growing seasons, and derived weather variables during the same years were analyzed using nonlinear and multiple regression analyses. Meteorological data were obtained from the Georgia Automated Environmental Monitoring Network. The best model explained 61% of the variation in spotted wilt severity (square root transformed) as a function of the interactions between the TSWV risk index, the average daily temperature in April (TavA), the average daily minimum temperature between March and April (TminMA), the accumulated rainfall in March (RainfallM), the accumulated rainfall in April (RainfallA), the number of rain days in April (RainDayA), evapotranspiration in April (EVTA), and the number of days from 1 January to the planting date (JulianDay). Integrating this weather-based model with the TSWV risk index may help peanut growers more effectively manage tomato spotted wilt disease. PMID:18943452

Olatinwo, R O; Paz, J O; Brown, S L; Kemerait, R C; Culbreath, A K; Beasley, J P; Hoogenboom, G

2008-10-01

359

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

UK PubMed Central (United Kingdom)

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

Geerts-Dimitriadou C; Lu YY; Geertsema C; Goldbach R; Kormelink R

2012-01-01

360

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

Scientific Electronic Library Online (English)

Full Text Available Abstract in portuguese Linhagens avançadas do programa de melhoramento do tomateiro (Lycopersicon esculentum) do IAC foram avaliadas em condições de campo em Campinas (SP) para resistência a tospovírus e a potyvírus, nos anos agrícolas 2002/2003 e 2003/2004, respectivamente. No primeiro ano, a única espécie de tospovírus que ocorreu na área experimental foi Tomato chlorotic spot virus (TCSV). As sete linhagens do grupo IAC exibiram baixa porcentagem de plantas sintomáticas em duas a (more) valiações, com médias abaixo de 28%; as cultivares testadas mostraram-se altamente suscetíveis, com médias acima de 85%, à exceção de 'Franco', que apresentou cerca de 55% de infecção. No segundo experimento, conduzido em 2003/2004, dez linhagens do grupo IAC foram comparadas com cinco cultivares de polinização aberta e híbridos F1, além do acesso LA-444-1 de L. peruvianum. Nesse experimento, por meio de testes biológicos e sorológicos, verificou-se ocorrência generalizada de Potato virus Y (PVY). Foi determinado o percentual de plantas com sintomas e avaliada a intensidade dos sintomas mediante uso de escala de notas. Com base nos dois critérios, verificou-se que LA-444-1 apresenta alta resistência a PVY, que 'Tyrade' exibe comportamento intermediário, enquanto todos os demais genótipos demonstram alta suscetibilidade ao vírus. O comportamento dos genótipos avaliados neste trabalho mostra a necessidade de se considerar, nos programas de melhoramento do tomateiro, a introgressão de fatores de resistência não só a vírus de importância atual nas regiões produtoras, como geminivírus, mas também a outros vírus potencialmente nocivos à cultura, como tospovírus e potyvírus. Abstract in english Advanced breeding lines from the IAC tomato breeding program and several tomato (Lycopersicon esculentum) cultivars and F1 hybrids were screened for tospovirus and potyvirus resistance under field conditions, at Campinas, São Paulo State, Brazil, during the 2002/2003 and 2003/2004 growing seasons. During the first season, only Tomato chlorotic spot virus (TCSV) was detected in plants of the experimental area. On both evaluations, all seven lines (IAC group) were resistan (more) t to TCSV, with under 28% of infected plants. The tomato cultivars and hybrids were highly susceptible, with greater than 85% of infected plants, except for 'Franco', with 55% of infected plants. During the 2003/2004 growing season, the number of IAC lines evaluated was raised from seven to ten, and they were compared to five cultivars/F1 hybrids and to L. peruvianum LA-444-1. On this experiment, only Potato virus Y (PVY) was detected in plants. Evaluations were carried out using a symptom intensity scale and ELISA. Considering both criteria, it was verified that only LA-444-1 displayed high resistance to PVY. In addition, 'Tyrade' displayed an intermediate behavior while all other lines, cultivars and hybrids behaved as susceptible to this potyvirus. These results highlight the need of introgressing resistance to multiple viruses in tomato breeding programs, taking into consideration the economical importance and relative incidence of each virus in different geographical regions and natural variations on incidence from year to year.

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

2005-12-01

 
 
 
 
361

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

Directory of Open Access Journals (Sweden)

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

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

2005-01-01

362

Dried blood spots as a practical and inexpensive source for human immunodeficiency virus and hepatitis C virus surveillance  

Scientific Electronic Library Online (English)

Full Text Available Abstract in english Passive surveillance of infectious diseases with a high percentage of asymptomatic cases or long incubation periods, such as acquired immunodeficiency syndrome (AIDS), does not reflect the current transmission dynamics. Thus, a multi-strategic surveillance, such as the human immunodeficiency virus (HIV) sentinel surveillance proposed by the World Health Organization (WHO), is necessary. The Brazilian HIV sentinel surveillance was started in May 1992 with this purpose. The (more) objectives of this study were to evaluate the feasibility and costs of HIV and hepatitis C virus (HCV) surveillance using dried blood spots (DBS) collected for neonatal screening of metabolic diseases in the state of Minas Gerais, Brazil. This was accomplished through the comparison of HIV and HCV seroprevalence with previous Brazilian studies. From December 2001 to June 2002, 24,905 newborns were tested for HIV and 4211 for HCV. HIV seroprevalence was 0.25% and the 95% confidence interval (CI) was 0.18, 0.31%; and HCV seroprevalence was 0.71% and the 95% CI was 0.46, 0.97%. These numbers are similar to previous Brazilian studies. Cost in this study was approximately US$ 3.10 per sample, which was roughly one third of the cost of the same exam at the Brazilian HIV sentinel surveillance. We conclude that it is possible and more cost-effective to use DBS for infectious diseases surveillance, albeit it is still necessary to compare these results with the usual sentinel methodology in a concomitant trial.

Toledo Jr., Antonio Carlos de Castro; Januário, José Nélio; Rezende, Renata Maria Silva; Siqueira, Arminda Lúcia; Mello, Bernardo Freire de; Fialho, Érica Ligorio; Ribeiro, Raquel Andrade; Silva, Hélia Lemos da; Pires, Érika Carvalho; Simões, Taynaná César; Greco, Dirceu Bartolomeu

2005-07-01

363

Dried blood spots as a practical and inexpensive source for human immunodeficiency virus and hepatitis C virus surveillance  

Directory of Open Access Journals (Sweden)

Full Text Available Passive surveillance of infectious diseases with a high percentage of asymptomatic cases or long incubation periods, such as acquired immunodeficiency syndrome (AIDS), does not reflect the current transmission dynamics. Thus, a multi-strategic surveillance, such as the human immunodeficiency virus (HIV) sentinel surveillance proposed by the World Health Organization (WHO), is necessary. The Brazilian HIV sentinel surveillance was started in May 1992 with this purpose. The objectives of this study were to evaluate the feasibility and costs of HIV and hepatitis C virus (HCV) surveillance using dried blood spots (DBS) collected for neonatal screening of metabolic diseases in the state of Minas Gerais, Brazil. This was accomplished through the comparison of HIV and HCV seroprevalence with previous Brazilian studies. From December 2001 to June 2002, 24,905 newborns were tested for HIV and 4211 for HCV. HIV seroprevalence was 0.25% and the 95% confidence interval (CI) was 0.18, 0.31%; and HCV seroprevalence was 0.71% and the 95% CI was 0.46, 0.97%. These numbers are similar to previous Brazilian studies. Cost in this study was approximately US$ 3.10 per sample, which was roughly one third of the cost of the same exam at the Brazilian HIV sentinel surveillance. We conclude that it is possible and more cost-effective to use DBS for infectious diseases surveillance, albeit it is still necessary to compare these results with the usual sentinel methodology in a concomitant trial.

Antonio Carlos de Castro Toledo Jr.; José Nélio Januário; Renata Maria Silva Rezende; Arminda Lúcia Siqueira; Bernardo Freire de Mello; Érica Ligorio Fialho; Raquel Andrade Ribeiro; Hélia Lemos da Silva; Érika Carvalho Pires; Taynaná César Simões; Dirceu Bartolomeu Greco

2005-01-01

364

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

Science.gov (United States)

Plant pathogens and insect herbivores are prone to share hosts under natural conditions. Consequently, pathogen-induced changes in the host plant can affect herbivory, and vice versa. Even though plant viruses are ubiquitous in the field, little is known about plant-mediated interactions between viruses and non-vectoring herbivores. We investigated the effects of virus infection on subsequent infestation by a non-vectoring herbivore in a natural genotype of Trifolium repens (white clover). We tested whether infection with White clover mosaic virus (WClMV) alters (1) the effects of fungus gnat feeding on plant growth, (2) the attractiveness of white clover for adult fungus gnat females, and (3) the volatile emission of white clover plants. We observed only marginal effects of WClMV infection on the interaction between fungus gnat larvae and white clover. However, adult fungus gnat females clearly preferred non-infected over WClMV-infected plants. Non-infected and virus-infected plants could easily be discriminated based on their volatile blends, suggesting that the preference of fungus gnats for non-infected plants may be mediated by virus-induced changes in volatile emissions. The compound ?-caryophyllene was exclusively detected in the headspace of virus-infected plants and may hence be particularly important for the preference of fungus gnat females. Our results demonstrate that WClMV infection can decrease the attractiveness of white clover plants for fungus gnat females. This suggests that virus infections may contribute to protecting their hosts by decreasing herbivore infestation rates. Consequently, it is conceivable that viruses play a more beneficial role in plant-herbivore interactions than generally thought. PMID:22526939

van Molken, Tamara; de Caluwe, Hannie; Hordijk, Cornelis A; Leon-Reyes, Antonio; Snoeren, Tjeerd A L; van Dam, Nicole M; Stuefer, Josef F

2012-04-17

365

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

UK PubMed Central (United Kingdom)

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

van Molken T; de Caluwe H; Hordijk CA; Leon-Reyes A; Snoeren TA; van Dam NM; Stuefer JF

2012-10-01

366

Genotyping of rubella virus RNA in sera and dried blood spots collected during routine surveillance and in archival sera.  

UK PubMed Central (United Kingdom)

Information on the molecular epidemiology of rubella has been valuable in supporting efforts to control and eliminate rubella in several countries. The preferred samples for virus isolation or RNA detection, such as throat swabs, are often not available making it difficult to obtain a robust database of rubella virus sequences. A method for obtaining rubella virus genotypes from more commonly collected samples such as sera or dried blood spots using real-time RT-PCR to screen samples followed by nested set amplification is described. Rubella genotypes were obtained from dried blood spots and recent and archival sera collections. Eighteen percent of the RNAs extracted from the archival sera were real-time RT-PCR positive, and 44% of these RNAs were amplified successfully by nested RT-PCR and sequenced. Implementation of this technique could provide another tool to improve global rubella molecular surveillance.

Zheng Q; Abernathy ES; Sun H; Zhu Z; de Filippis A; Akoua-Koffi C; Ahmed H; Morris-Glasgow V; Quist-Therson M; Icenogle JP

2013-02-01

367

Population genetic analysis of Tomato spotted wilt virus on peanut in North Carolina and Virginia.  

UK PubMed Central (United Kingdom)

Exploring the genetic diversity and evolutionary history of plant viruses is critical to understanding their ecology and epidemiology. In this study, maximum-likelihood and population genetics-based methods were used to investigate the population structure, genetic diversity, and sources of genetic variation in field isolates of Tomato spotted wilt virus (TSWV) from peanut in North Carolina and Virginia. Selected regions of the nucleocapsid, movement, and RNA-dependent RNA polymerase genes were amplified and sequenced to identify haplotypes and infer genetic relationships between isolates of TSWV with heuristic methods. The haplotype structure of each locus consisted of 1 or 2 predominant haplotypes and >100 haplotypes represented by a single isolate. No specific haplotypes were associated with geographic area, peanut cultivar, or year of isolation. The population was panmictic at the regional level and high levels of genetic diversity were observed among isolates. There was evidence for positive selection on single amino acids in each gene on a background of predominant purifying selection acting upon each locus. The results of compatibility analyses and the persistence of specific gene sequences in isolates collected over three field seasons suggest that recombination was occurring in the population. Estimates of the population mutation rate suggest that mutation has had a significant effect on the shaping of this population and, together with purifying selection, these forces have been the predominant evolutionary forces influencing the TSWV population in peanut in North Carolina and Virginia.

Kaye AC; Moyer JW; Parks EJ; Carbone I; Cubeta MA

2011-01-01

368

Viral genetic determinants for thrips transmission of Tomato spotted wilt virus.  

Science.gov (United States)

Tomato spotted wilt virus (TSWV) is transmitted exclusively by thrips in nature. A reassortment-based viral genetic system was used to map transmissibility by thrips to the medium (M) RNA of TSWV. To locate determinants of thrips transmission in the M RNA, 30 single-lesion isolates (SLIs) were generated from a single TSWV isolate that was inefficiently transmitted by thrips. Three of the 30 SLIs were transmitted by thrips, and 27 were not. Sequence analysis of the M RNA, thrips transmissibility assays, G(C) protein analysis, and transmission electron microscopic studies revealed that a specific nonsynonymous mutation (C1375A) in the G(N)/G(C) ORF of the M RNA resulted in the loss of thrips transmissibility without inhibition of virion assembly. This was in contrast to other nontransmissible SLIs, which had frameshift and/or nonsense mutations in the G(N)/G(C) ORF but were defective in virion assembly. The G(C) glycoprotein was detectable in the C1375A mutants but not in the frameshift or nonsense mutants. We report a specific viral determinant associated with virus transmission by thrips. In addition, the loss of transmissibility was associated with the accumulation of defective haplotypes in the population, which are not transmissible by thrips, rather than with the presence of a dominant haplotype that is inefficiently transmitted by thrips. These results also indicate that the glycoproteins may not be required for TSWV infection of plant hosts but are required for transmissibility by thrips. PMID:15753307

Sin, Sang-Hoon; McNulty, Brian C; Kennedy, George G; Moyer, James W

2005-03-07

369

Tomato spotted wilt virus glycoprotein G(C) is cleaved at acidic pH.  

Science.gov (United States)

Tomato spotted wilt virus (TSWV) is a plant-infecting member of the family Bunyaviridae. TSWV encodes two envelope glycoproteins, G(N) and G(C), which are required for virus infection of the arthropod vector. Other members of the Bunyaviridae enter host cells by pH-dependent endocytosis. During this process, the glycoproteins are exposed to conditions of acidic pH within endocytic vesicles causing the G(C) protein to change conformation. This conformational change renders G(C) more sensitive to protease cleavage. We subjected TSWV virions to varying pH conditions and determined that TSWV G(C), but not G(N), was cleaved under acidic pH conditions, and that this phenomenon did not occur at neutral or alkaline pH. This data provides evidence that G(C) changes conformation at low pH which results in altered protease sensitivity. Furthermore, sequence analysis of G(C) predicts the presence of internal hydrophobic domains, regions that are characteristic of fusion proteins. Like studies with other members of the Bunyaviridae, this study is the first step towards characterizing the nature of cell entry by TSWV. PMID:15845270

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

2005-06-01

370

Population genetic analysis of Tomato spotted wilt virus on peanut in North Carolina and Virginia.  

Science.gov (United States)

Exploring the genetic diversity and evolutionary history of plant viruses is critical to understanding their ecology and epidemiology. In this study, maximum-likelihood and population genetics-based methods were used to investigate the population structure, genetic diversity, and sources of genetic variation in field isolates of Tomato spotted wilt virus (TSWV) from peanut in North Carolina and Virginia. Selected regions of the nucleocapsid, movement, and RNA-dependent RNA polymerase genes were amplified and sequenced to identify haplotypes and infer genetic relationships between isolates of TSWV with heuristic methods. The haplotype structure of each locus consisted of 1 or 2 predominant haplotypes and >100 haplotypes represented by a single isolate. No specific haplotypes were associated with geographic area, peanut cultivar, or year of isolation. The population was panmictic at the regional level and high levels of genetic diversity were observed among isolates. There was evidence for positive selection on single amino acids in each gene on a background of predominant purifying selection acting upon each locus. The results of compatibility analyses and the persistence of specific gene sequences in isolates collected over three field seasons suggest that recombination was occurring in the population. Estimates of the population mutation rate suggest that mutation has had a significant effect on the shaping of this population and, together with purifying selection, these forces have been the predominant evolutionary forces influencing the TSWV population in peanut in North Carolina and Virginia. PMID:20839960

Kaye, A C; Moyer, J W; Parks, E J; Carbone, I; Cubeta, M A

2011-01-01

371

Identification of Dictyothrips betae as the vector of Polygonum ring spot virus  

UK PubMed Central (United Kingdom)

Dictyothrips betae (Thysanoptera: Thripidae) is the predominant thrips species on Polygonum convolvulus and Polygonum dumetorum plants infected with a recently described tospovirus species, Polygonum ring spot virus (PolRSV). Laboratory transmission experiments (leaf disk assays) with adults collected directly in the field demonstrated the competence of this thrips to transmit PolRSV, although only at a rate of 4%. However, this increased to 16% using newly emerged larvae fed on infected leaves. Frankliniella occidentalis and Thrips tabaci failed to transmit PolRSV in leaf disk assays. Reverse transcription-polymerase chain reaction (RT-PCR) with specific primers for the N protein and Western blot analysis of adult thrips to detect the N protein confirmed the presence of the virus in D. betae individuals after feeding for at least 5 days on healthy plants. For molecular identification purposes partial sequences of mitochondrial cytochrome c oxidase subunit I (COI), nuclear 28S ribosomal DNA and the elongation factor-1? (EF-1?) from D. betae were cloned. COI sequence was also used for deriving a phylogenetic tree, including D. betae. The results confirmed a relationship between this species and tospovirus-transmitting insects of the genus Thrips.

Ciuffo M; Mautino GC; Bosco L; Turina M; Tavella L

2010-09-01

372

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

UK PubMed Central (United Kingdom)

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

Whitfield AE; Ullman DE; German TL

2004-12-01

373

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

Science.gov (United States)

Tomato spotted wilt virus (TSWV), a member of the Tospovirus genus within the Bunyaviridae, is an economically important plant pathogen with a worldwide distribution. TSWV is transmitted to plants via thrips (Thysanoptera: Thripidae), which transmit the virus in a persistent propagative manner. The envelope glycoproteins, G(N) and G(C), are critical for the infection of thrips, but they are not required for the initial infection of plants. Thus, it is assumed that the envelope glycoproteins play important roles in the entry of TSWV into the insect midgut, the first site of infection. To directly test the hypothesis that G(N) plays a role in TSWV acquisition by thrips, we expressed and purified a soluble, recombinant form of the G(N) protein (G(N)-S). The expression of G(N)-S allowed us to examine the function of G(N) in the absence of other viral proteins. We detected specific binding to thrips midguts when purified G(N)-S was fed to thrips in an in vivo binding assay. The TSWV nucleocapsid protein and human cytomegalovirus glycoprotein B did not bind to thrips midguts, indicating that the G(N)-S-thrips midgut interaction is specific. TSWV acquisition inhibition assays revealed that thrips that were concomitantly fed purified TSWV and G(N)-S had reduced amounts of virus in their midguts compared to thrips that were fed TSWV only. Our findings that G(N)-S binds to larval thrips guts and decreases TSWV acquisition provide evidence that G(N) may serve as a viral ligand that mediates the attachment of TSWV to receptors displayed on the epithelial cells of the thrips midgut. PMID:15542672

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

2004-12-01

374

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

UK PubMed Central (United Kingdom)

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

Batuman O; Kuo YW; Palmieri M; Rojas MR; Gilbertson RL

2010-06-01

375

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

Science.gov (United States)

Tomatoes in Guatemala have been affected by a new disease, locally known as “mancha de chocolate” (chocolate spot). The disease is characterized by distinct necrotic spots on leaves, stems and petioles that eventually expand and cause a dieback of apical tissues. Samples from symptomatic plants tested negative for infection by tomato spotted wilt virus, tobacco streak virus, tobacco etch