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Sample records for maize white spot

  1. White-spot disease of salmon fry

    USGS Publications Warehouse

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

    1959-01-01

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

  2. Molecular and Ultrastructural Properties of Maize White Line Virus

    Technology Transfer Automated Retrieval System (TEKTRAN)

    This paper reports the complete nucleotide sequence of the genome of Maize white line mosaic virus (MWLMV) and describes the ultrastructural features of infected maize cells. The viral genome is an RNA molecule 4293 nt in size with the same structural organization of members of the Aureusvirus and ...

  3. Species of Cercospora associated with grey leaf spot of maize.

    PubMed

    Crous, Pedro W; Groenewald, Johannes Z; Groenewald, Marizeth; Caldwell, Pat; Braun, Uwe; Harrington, Thomas C

    2006-01-01

    Grey leaf spot is a serious yield-reducing disease of maize (Zea mays) in many parts of the world where this crop is cultivated. The causal organism associated with the disease is Cercospora zeae-maydis. Two potential sibling species have been recognized as Groups I and II. The DNA sequences for the internal transcribed spacers (ITS1 & ITS2), the 5.8S rRNA gene, elongation factor 1-alpha, histone H3, actin and calmodulin gene regions suggest that Groups I and II are two distinct species. Furthermore, Cercospora zeae-maydis (Group I) can be distinguished from C. zeina sp. nov. (Group II) by its faster growth rate on artificial media, the ability to produce cercosporin, longer conidiophores, and broadly fusiform conidia. A PCR-based test that distinguishes the two species was developed using species-specific primers designed from the histone H3 gene.

  4. Vibrio sp. causing Porites ulcerative white spot disease.

    PubMed

    Arboleda, Mark D M; Reichardt, Wolfgang T

    2010-06-11

    The causative agent of the Indo-Pacific coral disease, Porites ulcerative white spot syndrome (PUWS), that affects Porites spp. and a few other coral genera has so far remained unidentified. Inoculation of thiosulphate citrate bile sucrose (TCBS) agar with tissue material from Porites cylindrica infected with white spot produced colonies of approximately 3 mm diameter consisting of Gram-negative, motile, non-sucrose-fermenting, slightly curved rods with a minimum NaCl requirement of 0.3%. Three of these putative Vibrio sp. isolates were used for infection trials that included different stages of cell growth. Four modes of inoculation and 3 stages of bacterial cell growth were considered for testing Koch's postulates. Stationary phase cells proved more consistently infectious than did exponentially growing or starved cells using a 1-step immersion technique at cell concentrations of 10(4) cells ml(-1). A 1-step immersion technique proved more reliable in producing signs of white spot than did other techniques, such as injection, smearing and 2-step immersion of the inoculum. At inoculum densities >10(4) cells ml(-1) further signs of disease, such as tissue degradation and bleaching, also became evident. At elevated temperatures (>29 degrees C) bleaching remained absent for at least 2 mo from non-inoculated corals serving as controls, but was observed in artificially infected coral fragments. Of the 9 seawater aquaria containing healthy specimens of P. cylindrica, 6 showed signs of white spot 15 d after infection with an isolate tentatively identified as Vibrio sp. Based on 99% similarity of its 16S rRNA gene sequence and selected phenotypical features, this isolate revealed a close relationship to V. natriegens and V. parahaemolyticus.

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

    NASA Astrophysics Data System (ADS)

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

    2014-09-01

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

  6. A DARK SPOT ON A MASSIVE WHITE DWARF

    SciTech Connect

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

    2015-12-01

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

  7. A Dark Spot on a Massive White Dwarf

    NASA Astrophysics Data System (ADS)

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

    2015-12-01

    We present the serendipitous discovery of eclipse-like events around the massive white dwarf SDSS J152934.98+292801.9 (hereafter J1529+2928). We selected J1529+2928 for time-series photometry based on its spectroscopic temperature and surface gravity, which place it near the ZZ Ceti instability strip. Instead of pulsations, we detect photometric dips from this white dwarf every 38 minutes. Follow-up optical spectroscopy observations with Gemini reveal no significant radial velocity variations, ruling out stellar and brown dwarf companions. A disintegrating planet around this white dwarf cannot explain the observed light curves in different filters. Given the short period, the source of the photometric dips must be a dark spot that comes into view every 38 minutes due to the rotation of the white dwarf. Our optical spectroscopy does not show any evidence of Zeeman splitting of the Balmer lines, limiting the magnetic field strength to B < 70 kG. Since up to 15% of white dwarfs display kG magnetic fields, such eclipse-like events should be common around white dwarfs. We discuss the potential implications of this discovery on transient surveys targeting white dwarfs, like the K2 mission and the Large Synoptic Survey Telescope. This work is based on observations obtained at the Gemini Observatory, McDonald Observatory, and the Apache Point Observatory 3.5-m telescope. The latter is owned and operated by the Astrophysical Research Consortium. Gemini Observatory is operated by the Association of Universities for Research in Astronomy, Inc., under a cooperative agreement with the NSF on behalf of the Gemini partnership: the National Science Foundation (United States), the National Research Council (Canada), CONICYT (Chile), the Australian Research Council (Australia), Ministério da Ciência, Tecnologia e Inovação (Brazil) and Ministerio de Ciencia, Tecnología e Innovación Productiva (Argentina).

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

    PubMed Central

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

    2013-01-01

    The formation of white spot lesions or enamel demineralization around fixed orthodontic attachments is a common complication during and following fixed orthodontic treatment, which mars the result of a successfully completed case. This article is a contemporary review of the risk factors, preventive methods and fate of these orthodontics scars. The importance of excellent oral hygiene practice during fixed orthodontic treatment must be explained. Preventive programs must be emphasized to all orthodontic patients. Suggestions are offered in the literature for ways to prevent this condition from manifesting itself. PMID:24987641

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

    PubMed Central

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

    2014-01-01

    The Dominant White locus (W) in the domestic cat demonstrates pleiotropic effects exhibiting complete penetrance for absence of coat pigmentation and incomplete penetrance for deafness and iris hypopigmentation. We performed linkage analysis using a pedigree segregating White to identify KIT (Chr. B1) as the feline W locus. Segregation and sequence analysis of the KIT gene in two pedigrees (P1 and P2) revealed the remarkable retrotransposition and evolution of a feline endogenous retrovirus (FERV1) as responsible for two distinct phenotypes of the W locus, Dominant White, and white spotting. A full-length (7125 bp) FERV1 element is associated with white spotting, whereas a FERV1 long terminal repeat (LTR) is associated with all Dominant White individuals. For purposes of statistical analysis, the alternatives of wild-type sequence, FERV1 element, and LTR-only define a triallelic marker. Taking into account pedigree relationships, deafness is genetically linked and associated with this marker; estimated P values for association are in the range of 0.007 to 0.10. The retrotransposition interrupts a DNAase I hypersensitive site in KIT intron 1 that is highly conserved across mammals and was previously demonstrated to regulate temporal and tissue-specific expression of KIT in murine hematopoietic and melanocytic cells. A large-population genetic survey of cats (n = 270), representing 30 cat breeds, supports our findings and demonstrates statistical significance of the FERV1 LTR and full-length element with Dominant White/blue iris (P < 0.0001) and white spotting (P < 0.0001), respectively. PMID:25085922

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

    PubMed

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

    2014-08-01

    The Dominant White locus (W) in the domestic cat demonstrates pleiotropic effects exhibiting complete penetrance for absence of coat pigmentation and incomplete penetrance for deafness and iris hypopigmentation. We performed linkage analysis using a pedigree segregating White to identify KIT (Chr. B1) as the feline W locus. Segregation and sequence analysis of the KIT gene in two pedigrees (P1 and P2) revealed the remarkable retrotransposition and evolution of a feline endogenous retrovirus (FERV1) as responsible for two distinct phenotypes of the W locus, Dominant White, and white spotting. A full-length (7125 bp) FERV1 element is associated with white spotting, whereas a FERV1 long terminal repeat (LTR) is associated with all Dominant White individuals. For purposes of statistical analysis, the alternatives of wild-type sequence, FERV1 element, and LTR-only define a triallelic marker. Taking into account pedigree relationships, deafness is genetically linked and associated with this marker; estimated P values for association are in the range of 0.007 to 0.10. The retrotransposition interrupts a DNAase I hypersensitive site in KIT intron 1 that is highly conserved across mammals and was previously demonstrated to regulate temporal and tissue-specific expression of KIT in murine hematopoietic and melanocytic cells. A large-population genetic survey of cats (n = 270), representing 30 cat breeds, supports our findings and demonstrates statistical significance of the FERV1 LTR and full-length element with Dominant White/blue iris (P < 0.0001) and white spotting (P < 0.0001), respectively.

  11. Systems genetics reveals a transcriptional network associated with susceptibility in the maize-grey leaf spot pathosystem

    Technology Transfer Automated Retrieval System (TEKTRAN)

    We have applied a systems genetics approach to elucidate molecular mechanisms of maize responses to gray leaf spot (GLS) disease, caused by Cercospora zeina, a major threat to maize production globally. We conducted expression QTL (eQTL) analysis of gene expression variation measured in earleaf samp...

  12. Inducibility of somatic colour and white spots in the mammalian spot test.

    PubMed

    Peter, S

    1980-03-01

    F1 embryos of the C57BL/6JHan x T-Stock were exposed to utero to 5, 10, 15 and 45 mg/kg b.w. of cyclophosphamide (CP) s.c. on the 10th day of pregnancy. 3-5 weeks post-partum the offspring were examined for the appearance of recessive coat-colour mosaics and white midventral spots (WMVS). The frequencies of the coloured spots were 0.93 and 2.59% at 5- and 10-mg/kg doses of CP, resp. These frequencies were dose-dependent, but the difference between the doses was not significant. No coloured spots were obtained with CP at 15 and 45 mg/kg. The frequencies of WMVS were much higher than those of coloured spots (1.40, 6.03 and 51.16%, resp.). The differences were highly significant. The reduction in the number of offspring compared with the raio of the offspring/female of the control (control = 100%) were 16.16, 22.72, 72.48 and 100% at the different doses of CP (5, 10, 15 and 45 mg/kg b.w.).

  13. Complete Genome Sequence of the Shrimp White Spot Bacilliform Virus

    PubMed Central

    Yang, Feng; He, Jun; Lin, Xionghui; Li, Qin; Pan, Deng; Zhang, Xiaobo; Xu, Xun

    2001-01-01

    We report the first complete genome sequence of a marine invertebrate virus. White spot bacilliform virus (WSBV; or white spot syndrome virus) is a major shrimp pathogen with a high mortality rate and a wide host range. Its double-stranded circular DNA genome of 305,107 bp contains 181 open reading frames (ORFs). Nine homologous regions containing 47 repeated minifragments that include direct repeats, atypical inverted repeat sequences, and imperfect palindromes were identified. This is the largest animal virus that has been completely sequenced. Although WSBV is morphologically similar to insect baculovirus, the two viruses are not detectably related at the amino acid level. Rather, some WSBV genes are more homologous to eukaryotic genes than viral genes. In fact, sequence analysis indicates that WSBV differs from all known viruses, although a few genes display a weak homology to herpesvirus genes. Most of the ORFs encode proteins that bear no homology to any known proteins, either suggesting that WSBV represents a novel class of viruses or perhaps implying a significant evolutionary distance between marine and terrestrial viruses. The most unique feature of WSBV is the presence of an intact collagen gene, a gene encoding an extracellular matrix protein of animal cells that has never been found in any viruses. Determination of the genome of WSBV will facilitate a better understanding of the molecular mechanism underlying the pathogenesis of the WSBV virus and will also provide useful information concerning the evolution and divergence of marine and terrestrial animal viruses at the molecular level. PMID:11689662

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

    PubMed Central

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

    2013-01-01

    This article describes the treatment provided to a patient with the maxillary anterior teeth exhibiting severe secondary caries beneath the previous restoration and a white spot lesion on the adjacent incisor. Two implants were placed after extraction of hopeless teeth with the guided bone regeneration technique. A white spot lesion of the adjacent incisor was treated with minimally invasive treatment. This clinical report describes the multidisciplinary treatment for the white spot lesion and esthetic restoration of missing anterior teeth. PMID:24049579

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

    PubMed Central

    Sánchez-Paz, Arturo

    2010-01-01

    Viruses are ubiquitous and extremely abundant in the marine environment. One of such marine viruses, the white spot syndrome virus (WSSV), has emerged globally as one of the most prevalent, widespread and lethal for shrimp populations. However, at present there is no treatment available to interfere with the unrestrained occurrence and spread of the disease. The recent progress in molecular biology techniques has made it possible to obtain information on the factors, mechanisms and strategies used by this virus to infect and replicate in susceptible host cells. Yet, further research is still required to fully understand the basic nature of WSSV, its exact life cycle and mode of infection. This information will expand our knowledge and may contribute to developing effective prophylactic or therapeutic measures. This review provides a state-of-the-art overview of the topic, and emphasizes the current progress and future direction for the development of WSSV control strategies. PMID:20181325

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

    PubMed

    Sánchez-Paz, Arturo

    2010-01-01

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

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

    PubMed Central

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

    2016-01-01

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

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

    PubMed

    Rodriguez-Anaya, Libia Zulema; Gonzalez-Galaviz, Jose Reyes; Casillas-Hernandez, Ramón; Lares-Villa, Fernando; Estrada, Karel; Ibarra-Gamez, Jose Cuauhtemoc; Sanchez-Flores, Alejandro

    2016-03-10

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

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

    PubMed

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

    2013-12-01

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

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

    PubMed Central

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

    2017-01-01

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

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

    PubMed

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

    2012-09-01

    White-spot lesions (WSL) associated with orthodontic appliances are a cosmetic problem and increase risk for cavities. We characterized the microbiota of WSL, accounting for confounding due to gingivitis. Participants were 60 children with fixed appliances, aged between 10 and 19 yrs, half with WSL. Plaque samples were assayed by a 16S rRNA-based microarray (HOMIM) and by PCR. Mean gingival index was positively associated with WSL (p = 0.018). Taxa associated with WSL by microarray included Granulicatella elegans (p = 0.01), Veillonellaceae sp. HOT 155 (p < 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.

  2. Molecular epidemiology of white spot syndrome virus within Vietnam.

    PubMed

    Dieu, Bui Thi Minh; Marks, Hendrik; Siebenga, J Joukje; Goldbach, Rob W; Zuidema, Douwe; Duong, Tran Phuoc; Vlak, Just M

    2004-12-01

    White spot syndrome virus (WSSV), the sole member of the virus family Nimaviridae, is a large double-stranded DNA virus that infects shrimp and other crustaceans. By alignment of three completely sequenced isolates originating from Taiwan (WSSV-TW), China (WSSV-CN) and Thailand (WSSV-TH), the variable loci in the genome were mapped. The variation suggests the spread of WSSV from a common ancestor originating from either side of the Taiwan Strait to Thailand, but support for this hypothesis through analysis of geographical intermediates is sought. RFLP analysis of eight Vietnamese WSSV isolates, of which six were collected along the central coast (VN-central) and two along the south coast (VN-south), showed apparent sequence variation in the variable loci identified previously. These loci were characterized in detail by PCR amplification, cloning and sequencing. Relative to WSSV-TW, all VN-central isolates showed a approximately 8.5 kb deletion in the major variable region ORF23/24, whereas the VN-south isolates contain a deletion of approximately 11.5 or approximately 12.2 kb, compared to a approximately 1.2 or approximately 13.2 kb deletion in WSSV-CN and WSSV-TH, respectively. The minor variable region ORF14/15 showed deletions of various sizes compared with WSSV-TH for all eight VN isolates. The data suggest that the VN isolates and WSSV-TH have a common lineage, which branched off from WSSV-TW and WSSV-CN early on, and that WSSV entered Vietnam by multiple introductions. A model is presented for the spread of WSSV from either side of the Taiwan Strait into Vietnam based on the gradually increasing deletions of both 'variable regions'. The number and order of repeat units within ORF75 and ORF125 appeared to be suitable markers to study regional spread of WSSV.

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

    PubMed

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

    2015-03-06

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

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

    PubMed

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

    2016-01-01

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

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

    PubMed

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

    2010-12-01

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

  6. Maize white seedling 3 results from disruption of homogentisate solanesyl transferase

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Maize white seedling 3 (w3) has served as a model albino-seedling mutant since its discovery in 1923. We show here that the w3 phenotype is caused by disruptions in homogentisate solanesyl transferase (HST), an enzyme that catalyzes the committed step in plastoquinone-9 (PQ9) biosynthesis. This re...

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

    PubMed

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

    2015-05-01

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

  8. A de novo mutation in KIT causes white spotting in a subpopulation of German Shepherd dogs.

    PubMed

    Wong, A K; Ruhe, A L; Robertson, K R; Loew, E R; Williams, D C; Neff, M W

    2013-06-01

    Although variation in the KIT gene is a common cause of white spotting among domesticated animals, KIT has not been implicated in the diverse white spotting observed in the dog. Here, we show that a loss-of-function mutation in KIT recapitulates the coat color phenotypes observed in other species. A spontaneous white spotting observed in a pedigree of German Shepherd dogs was mapped by linkage analysis to a single locus on CFA13 containing KIT (pairwise LOD = 15). DNA sequence analysis identified a novel 1-bp insertion in the second exon that co-segregated with the phenotype. The expected frameshift and resulting premature stop codons predicted a severely truncated c-Kit receptor with presumably abolished activity. No dogs homozygous for the mutation were recovered from multiple intercrosses (P = 0.01), suggesting the mutation is recessively embryonic lethal. These observations are consistent with the effects of null alleles of KIT in other species.

  9. Handling and aflatoxin contamination of white maize in Costa Rica.

    PubMed

    Mora, M; Lacey, J

    1997-01-01

    Projects funded by International Development Research Centre (IDRC) of Canada and the European Commission have enabled the examination of more than 3000 samples of maize collected from all regions of Costa Rica at different stages, from the growing crop through storage to final sale, and at different water contents. Contamination with Aspergillus flavus was frequent and about 80% of samples contained more than 20 ng aflatoxins g(-1) grain. Average contamination with aflatoxins in the Brunca Region was > 274 ng g(-1) while that in other regions was < 70 ng g(-1). Except in Brunca region, where it averaged 376 ng g(-1), contamination of grain from commercial sources was slightly less than of that from farms (< or = 15 ng g(-1)). It appeared that samples kept on the cob after harvest contained almost no aflatoxin while shelled samples were frequently highly contaminated. Experiments were therefore done in Brunca and Huetar Atlantic Regions, utilising 34 experimental maize crops to study in detail the development of A. flavus and aflatoxin from before harvest, through postharvest treatment before drying and through storage for six months. A. flavus was isolated more frequently from maize shelled immediately after harvest than from that kept on the cob until it could be dried, and from more samples from the Brunca Region than from the Huetar Atlantic Region. Samples harvested with > or = 18% water content often contains > 70% of grains infected with A. flavus but sometimes there were few grains infected. As found in the initial survey, more aflatoxin contamination developed in shelled maize than in that handled on the cob during the period from harvesting to drying, especially if the delay was more than 5 days, and more in Brunca than in Huetar. Shelled grain contained 400-800 ng aflatoxin g(-1) in Brunca but < 100 ng g(-1) in Huetar while grain kept on the cob contained < 30 ng g(-1), even with > 18% water content. Incidence of Fusarium spp. exceeded 50% except where A

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

    PubMed

    Haase, B; Rieder, S; Leeb, T

    2015-06-01

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

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

    PubMed

    Chai, Choong Yee; Yoon, Jangmi; Lee, Yong Seok; Kim, Young Bong; Choi, Tae-Jin

    2013-10-01

    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.

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

    PubMed

    Kose; Mänd; Møller

    1999-12-01

    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.

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

    PubMed Central

    Jahanbin, Arezoo; Ameri, Hamideh; Shahabi, Mostafa; Ghazi, Ala

    2015-01-01

    Statement of the Problem Demineralization of enamel adjacent to orthodontic appliances frequently occurs, commonly due to insufficient oral hygiene. Purpose The aim of this study was to compare two microabrasion techniques on improving the white spot lesions as well as subsequent enamel discoloration. Materials and Method Sixty extracted premolar teeth without caries and hypoplasia were selected for this study. White spot lesions were artificially induced on the buccal surface of each tooth. Teeth were randomly assigned to three treatment groups, each treated with pumice powder as the control, microabrasion with 18% HCl, and microabrasion with 37% H3PO4. Subsequently, the three groups were daily immersed for five minutes in a tea-coffee solution for a period of one week. Colorimetric evaluation was done before and after formation of white spot lesions, after microabrasion, and after immersion in the colored solution; then the color differences (∆E) were calculated. Statistical analysis was performed by multiple measurement analysis and the Tukey’s test. Results This study showed that ∆E between the stages of white spot formation and microabrasion for H3PO4 was more than other groups and for the pumice powder group it was less than the others. Furthermore, there was a significant difference between ∆E of the three study groups (p= 0.017). Additionally, ∆E after placing the teeth in the colored solution and microabrasion was the highest for the HCl group and the lowest for the pumice powder group. There was also a significant difference between the three groups (p= 0.000). Conclusion Pumice powder alone had similar effects as 18% HCl on removing the white spot lesions. Nevertheless, 18% HCl makes the enamel susceptible for subsequent color staining more than the other microabrasion methods. PMID:26106636

  14. Physicochemical characterization of white, yellow and purple maize flours and rheological characterization of their doughs.

    PubMed

    Moreira, R; Chenlo, F; Arufe, S; Rubinos, S N

    2015-12-01

    White, yellow and purple maize flours were obtained after dried kernels milling with two different sieves (200 and 500 μm). Hygroscopic characteristics, particle size distribution, colour and total starch and damaged starch (DS) of flours were determined. Maize flour doughs were obtained by mixing of flour and water in a laboratory kneader (Mixolab®) at constant dough consistency (1.10 ± 0.07 Nm). Dough properties like water absorption (WA), development and stability times were determined. Rheological characterization was carried out at 30 °C by means of oscillatory frequency sweep (1-100 rad s(-1)) at 0.1 % strain and creep (50 Pa, 60 s) - recovery (0 Pa, 180 s) tests using a controlled stress rheometer. No significant differences were observed among water desorption isotherms of maize varieties and Halsey model was satisfactorily employed. Under the same milling conditions, white maize flours showed higher average particles size than purple and yellow maize flours. A model to predict flours colour involving colour parameters of the particle size fractions is proposed. Flours obtained with smaller particle size showed higher DS content and WA. For tested doughs, the mechanical spectra showed that elastic component was dominant over the viscous one. Damping factor varied slightly with angular frequency. Moduli values depended on average particle size and WA of dough. Creep-recovery data were satisfactorily fit with Burgers model. Instantaneous creep compliance varied with the same trend than elastic modulus. Viscoelastic creep compliance increased linearly with WA of the tested doughs and, at constant average flour particle size, increased with increasing DS.

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

    PubMed

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

    2015-02-01

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

  16. Pathology Associated with White Spot Virus (WSV) Infection in Wild Broodstock of Tiger Prawns (Penaeus monodon)

    PubMed Central

    Kua, Beng Chu; Rashid, Noraziah Mat

    2012-01-01

    A total of six wild broodstocks of tiger prawns, Penaeus monodon, were found positive for White Spot Virus (WSV) with an IQ2000 detection kit. Using histopathology, the intranuclear inclusion of haemocyte due to WSV infection was observed in the epithelium cells of the antennal gland, stomach and gills. This result confirmed that the wild broodstocks were positive with WSV without showing any white spot. Additionally, histopathological examination also revealed an accumulation of haemocytes around the hepatopancreatic tubules resulting from bacterial infection. Encapsulation and nodule formation, as well as related necrosis, were also observed around the hepatopancreatic tubules infected with a metazoan parasite. Encysted tylocephalum larval cestodes were observed in the hepatopancreas, with haemocytic aggregation being observed around the infected tubules. These findings showed some bacterial and parasitic infections which, in addition to the viral infection itself, could contribute to the 80% mortality rate in wild broodstocks infected with WSV. PMID:24575228

  17. Pathology Associated with White Spot Virus (WSV) Infection in Wild Broodstock of Tiger Prawns (Penaeus monodon).

    PubMed

    Kua, Beng Chu; Rashid, Noraziah Mat

    2012-05-01

    A total of six wild broodstocks of tiger prawns, Penaeus monodon, were found positive for White Spot Virus (WSV) with an IQ2000 detection kit. Using histopathology, the intranuclear inclusion of haemocyte due to WSV infection was observed in the epithelium cells of the antennal gland, stomach and gills. This result confirmed that the wild broodstocks were positive with WSV without showing any white spot. Additionally, histopathological examination also revealed an accumulation of haemocytes around the hepatopancreatic tubules resulting from bacterial infection. Encapsulation and nodule formation, as well as related necrosis, were also observed around the hepatopancreatic tubules infected with a metazoan parasite. Encysted tylocephalum larval cestodes were observed in the hepatopancreas, with haemocytic aggregation being observed around the infected tubules. These findings showed some bacterial and parasitic infections which, in addition to the viral infection itself, could contribute to the 80% mortality rate in wild broodstocks infected with WSV.

  18. Conservative approach for a clinical resolution of enamel white spot lesions.

    PubMed

    Nahsan, Flavia Pardo Salata; da Silva, Luciana Mendonça; Baseggio, Wagner; Franco, Eduardo Batista; Francisconi, Paulo Afonso Silveira; Mondelli, Rafael Francisco Lia; Wang, Linda

    2011-05-01

    Enamel white spot lesions in anterior teeth that compromise esthetics are common. Microabrasion is indicated, since it affects enamel superficially. An acid-abrasive slurry with 37% phosphoric acid with pumice was used on the enamel for a controlled time period. Home bleaching with hydrogen peroxide was then used, further improving the final result. The method is safe, easy, and conservative and provides good esthetic results.

  19. A simple repeat polymorphism in the MITF-M promoter is a key regulator of white spotting in dogs.

    PubMed

    Baranowska Körberg, Izabella; Sundström, Elisabeth; Meadows, Jennifer R S; Rosengren Pielberg, Gerli; Gustafson, Ulla; Hedhammar, Åke; Karlsson, Elinor K; Seddon, Jennifer; Söderberg, Arne; Vilà, Carles; Zhang, Xiaolan; Åkesson, Mikael; Lindblad-Toh, Kerstin; Andersson, Göran; Andersson, Leif

    2014-01-01

    The white spotting locus (S) in dogs is colocalized with the MITF (microphtalmia-associated transcription factor) gene. The phenotypic effects of the four S alleles range from solid colour (S) to extreme white spotting (s(w)). We have investigated four candidate mutations associated with the s(w) allele, a SINE insertion, a SNP at a conserved site and a simple repeat polymorphism all associated with the MITF-M promoter as well as a 12 base pair deletion in exon 1B. The variants associated with white spotting at all four loci were also found among wolves and we conclude that none of these could be a sole causal mutation, at least not for extreme white spotting. We propose that the three canine white spotting alleles are not caused by three independent mutations but represent haplotype effects due to different combinations of causal polymorphisms. The simple repeat polymorphism showed extensive diversity both in dogs and wolves, and allele-sharing was common between wolves and white spotted dogs but was non-existent between solid and spotted dogs as well as between wolves and solid dogs. This finding was unexpected as Solid is assumed to be the wild-type allele. The data indicate that the simple repeat polymorphism has been a target for selection during dog domestication and breed formation. We also evaluated the significance of the three MITF-M associated polymorphisms with a Luciferase assay, and found conclusive evidence that the simple repeat polymorphism affects promoter activity. Three alleles associated with white spotting gave consistently lower promoter activity compared with the allele associated with solid colour. We propose that the simple repeat polymorphism affects cooperativity between transcription factors binding on either flanking sides of the repeat. Thus, both genetic and functional evidence show that the simple repeat polymorphism is a key regulator of white spotting in dogs.

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

    PubMed

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

    2009-12-01

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

  1. Robust Identification of Binding Hot Spots Using Continuum Electrostatics: Application to Hen Egg-White Lysozyme

    PubMed Central

    2011-01-01

    Binding hot spots, protein regions with high binding affinity, can be identified by using X-ray crystallography or NMR spectroscopy to screen libraries of small organic molecules that tend to cluster at such hot spots. FTMap, a direct computational analogue of the experimental screening approaches, uses 16 different probe molecules for global sampling of the surface of a target protein on a dense grid and evaluates the energy of interaction using an empirical energy function that includes a continuum electrostatic term. Energy evaluation is based on the fast Fourier transform correlation approach, which allows for the sampling of billions of probe positions. The grid sampling is followed by off-grid minimization that uses a more detailed energy expression with a continuum electrostatics term. FTMap identifies the hot spots as consensus clusters formed by overlapping clusters of several probes. The hot spots are ranked on the basis of the number of probe clusters, which predicts their binding propensity. We applied FTMap to nine structures of hen egg-white lysozyme (HEWL), whose hot spots have been extensively studied by both experimental and computational methods. FTMap found the primary hot spot in site C of all nine structures, in spite of conformational differences. In addition, secondary hot spots in sites B and D that are known to be important for the binding of polysaccharide substrates were found. The predicted probe–protein interactions agree well with those seen in the complexes of HEWL with various ligands and also agree with an NMR-based study of HEWL in aqueous solutions of eight organic solvents. We argue that FTMap provides more complete information on the HEWL binding site than previous computational methods and yields fewer false-positive binding locations than the X-ray structures of HEWL from crystals soaked in organic solvents. PMID:22092261

  2. Systems genetics reveals a transcriptional network associated with susceptibility in the maize-grey leaf spot pathosystem.

    PubMed

    Christie, Nanette; Myburg, Alexander A; Joubert, Fourie; Murray, Shane L; Carstens, Maryke; Lin, Yao-Cheng; Meyer, Jacqueline; Crampton, Bridget G; Christensen, Shawn A; Ntuli, Jean F; Wighard, Sara S; Van de Peer, Yves; Berger, Dave K

    2017-02-01

    We used a systems genetics approach to elucidate the molecular mechanisms of the responses of maize to grey leaf spot (GLS) disease caused by Cercospora zeina, a threat to maize production globally. Expression analysis of earleaf samples in a subtropical maize recombinant inbred line population (CML444 × SC Malawi) subjected in the field to C. zeina infection allowed detection of 20 206 expression quantitative trait loci (eQTLs). Four trans-eQTL hotspots coincided with GLS disease QTLs mapped in the same field experiment. Co-expression network analysis identified three expression modules correlated with GLS disease scores. The module (GY-s) most highly correlated with susceptibility (r = 0.71; 179 genes) was enriched for the glyoxylate pathway, lipid metabolism, diterpenoid biosynthesis and responses to pathogen molecules such as chitin. The GY-s module was enriched for genes with trans-eQTLs in hotspots on chromosomes 9 and 10, which also coincided with phenotypic QTLs for susceptibility to GLS. This transcriptional network has significant overlap with the GLS susceptibility response of maize line B73, and may reflect pathogen manipulation for nutrient acquisition and/or unsuccessful defence responses, such as kauralexin production by the diterpenoid biosynthesis pathway. The co-expression module that correlated best with resistance (TQ-r; 1498 genes) was enriched for genes with trans-eQTLs in hotspots coinciding with GLS resistance QTLs on chromosome 9. Jasmonate responses were implicated in resistance to GLS through co-expression of COI1 and enrichment of genes with the Gene Ontology term 'cullin-RING ubiquitin ligase complex' in the TQ-r module. Consistent with this, JAZ repressor expression was highly correlated with the severity of GLS disease in the GY-s susceptibility network.

  3. Identification of alleles conferring resistance to gray leaf spot in maize derived from its wild progenitor species teosinte (Zea mays ssp. parviglumis)

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Gray Leaf Spot [(GLS), causal agent Cercospora zeae-maydis and Cercospora zeina] is an important maize disease in the United States. Current control methods for GLS include using resistant cultivars, crop rotation, chemical applications, and conventional tillage to reduce inoculum levels. Teosinte ...

  4. Enzyme E2 from Chinese white shrimp inhibits replication of white spot syndrome virus and ubiquitinates its RING domain proteins.

    PubMed

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

    2011-08-01

    Recent studies have shown that the ubiquitin (Ub) proteasome pathway (UPP) is closely related to immune defense. We have identified a ubiquitin-conjugating enzyme, E2, from the Chinese white shrimp, Fenneropenaeus chinensis (FcUbc). Injection of recombinant FcUbc protein (rFcUbc) reduced the mortality of shrimp infected with white spot syndrome virus (WSSV) and inhibited replication of WSSV. rFcUbc, but not a mutant FcUbc (mFcUbc), bound to WSSV RING domains (WRDs) from four potential E3 ligase proteins of WSSV in vitro. Importantly, rFcUbc could ubiquitinate the RING domains (named WRD2 and WRD3) of WSSV277 and WSSV304 proteins in vitro and the two proteins in WSSV-infected Drosophila melanogaster Schneider 2 (S2) cells. Furthermore, overexpression of FcUbc increased ubiquitination of WSSV277 and WSSV304 during WSSV infection. In summary, our study demonstrates that FcUbc from Chinese white shrimp inhibited WSSV replication and could ubiquitinate WSSV RING domain-containing proteins. This is the first report about antiviral function of Ubc E2 in shrimp.

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

    NASA Technical Reports Server (NTRS)

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

    1981-01-01

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

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

    NASA Astrophysics Data System (ADS)

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

    2000-09-01

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

  7. Mutations in MITF and PAX3 Cause “Splashed White” and Other White Spotting Phenotypes in Horses

    PubMed Central

    Blatter, Marlis; Brooks, Samantha A.; Burger, Dominik; Drögemüller, Cord; Gerber, Vincent; Henke, Diana; Janda, Jozef; Jude, Rony; Magdesian, K. Gary; Matthews, Jacqueline M.; Poncet, Pierre-André; Svansson, Vilhjálmur; Tozaki, Teruaki; Wilkinson-White, Lorna; Penedo, M. Cecilia T.; Rieder, Stefan; Leeb, Tosso

    2012-01-01

    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. PMID:22511888

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

    PubMed Central

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

    2014-01-01

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

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

    PubMed

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

    2016-01-18

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

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

    PubMed

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

    2014-04-10

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

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

    PubMed

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

    2014-06-01

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

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

    PubMed

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

    2015-02-01

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

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

    PubMed Central

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

    2016-01-01

    Since its emergence in the 1990s, White Spot Disease (WSD) has had major economic and societal impact in the crustacean aquaculture sector. Over the years shrimp farming alone has experienced billion dollar losses through WSD. The disease is caused by the White Spot Syndrome Virus (WSSV), a large dsDNA virus and the only member of the Nimaviridae family. Susceptibility to WSSV in a wide range of crustacean hosts makes it a major risk factor in the translocation of live animals and in commodity products. Currently there are no effective treatments for this disease. Understanding the molecular basis of disease processes has contributed significantly to the treatment of many human and animal pathogens, and with a similar aim considerable efforts have been directed towards understanding host–pathogen molecular interactions for WSD. Work on the molecular mechanisms of pathogenesis in aquatic crustaceans has been restricted by a lack of sequenced and annotated genomes for host species. Nevertheless, some of the key host–pathogen interactions have been established: between viral envelope proteins and host cell receptors at initiation of infection, involvement of various immune system pathways in response to WSSV, and the roles of various host and virus miRNAs in mitigation or progression of disease. Despite these advances, many fundamental knowledge gaps remain; for example, the roles of the majority of WSSV proteins are still unknown. In this review we assess current knowledge of how WSSV infects and replicates in its host, and critique strategies for WSD treatment. PMID:26797629

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

    PubMed

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

    2000-02-01

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

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

    PubMed Central

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

    2016-01-01

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

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

    SciTech Connect

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

    2006-08-01

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

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

    PubMed

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

    2016-05-01

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

  18. [Fatty acids profile characterization of white maize hybrids grown in Venezuela].

    PubMed

    Alezones, Jesús; Avila, Manuel; Chassaigne, Alberto; Barrientos, Venancio

    2010-12-01

    In Venezuela, white corn is the most important crop regarding production, harvest area and consumption. One of its main by-products is corn oil, whose positive effect on health caused by the high content of unsaturated fatty acids has been widely recognized. In order to characterize the fatty acids profile of twelve white grained maize hybrids extensively grown in Venezuela, and the effect that divergent localities has on this profile, three semi commercial scale trials where established in Portuguesa, Yaracuy and Guárico states. Proportions of the main fatty acids in the raw oil of the different grain samples were determined using gas chromatography. Significant differences (p < 0,01) between hybrids were found for arachidic, palmitic, stearic, oleic, gadoleic and linoleic acids; non significant differences were found for linolenic acid. Significant differences between localities were found for all the fatty acids evaluated. High and significant correlations between fatty acids content were found; the most important relations were: linoleic-oleic (Rho = -0,98**), arachidic-palmitic (Rho = -0,61**), linoleic-stearic (Rho = -0,61**) and oleic-stearic (Rho = 0,58**). Corn produced in Venezuela presents lower levels of linoleic and higher levels of palmitic, stearic and oleic acids than the levels found in temperate corn. These differences involve significant changes in the nutritional properties of Venezuelan corn oil that should be considered in the development of new cultivars and industrial processes for oil production.

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

    PubMed

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

    2012-04-26

    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.

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

    PubMed

    Nunan, Linda M; Lightner, Donald V

    2011-01-01

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

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

    PubMed

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

    2014-07-01

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

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

    PubMed

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

    2013-09-01

    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-β-cyclodextrin (MβCD) and chlorpromazine (CPZ). WSSV still can be detected in the hemocytes treated with CPZ, but not in the hemocytes treated with MβCD. Thus, we conclude that WSSV adopts the caveolae-mediated endocytosis to enter the shrimp cell.

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

    PubMed

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

    2015-02-01

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

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

    PubMed

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

    2015-12-01

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

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

    PubMed

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

    2010-12-20

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

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

    PubMed

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

    2013-10-01

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

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

    SciTech Connect

    Li Li; Lin Shumei; Yanga Feng . E-mail: mbiotech@public.xm.fj.cn

    2005-07-05

    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.

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

    PubMed

    Zhu, Fei; Jin, Min

    2015-12-01

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

  9. Rapid detection of shrimp white spot syndrome virus by real time, isothermal recombinase polymerase amplification assay.

    PubMed

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

    2014-01-01

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

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

    SciTech Connect

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

    2006-01-01

    The nonstructural protein VP9 from white spot syndrome virus (WSSV) has been identified and expressed in Escherichia coli. To facilitate purification, a cleavable 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.

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

    NASA Astrophysics Data System (ADS)

    Podray, Susan S.

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

  12. Expression, purification and crystallization of two major envelope proteins from white spot syndrome virus

    SciTech Connect

    Tang, Xuhua; Hew, Choy Leong

    2007-07-01

    The crystallization of the N-terminal transmembrane region-truncated VP26 and VP28 of white spot syndrome virus is described. 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 vapour-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 Å resolution and belong to space group R32, with unit-cell parameters a = b = 73.92, c = 199.31 Å. 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 Å, and diffracts to 2.0 Å resolution.

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

    PubMed Central

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

    2017-01-01

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

  14. Antibacterial and protein-repellent orthodontic cement to combat biofilms and white spot lesions

    PubMed Central

    Zhang, Ning; Chen, Chen; Weir, Michael D.; Bai, Yuxing; Xu, Hockin H. K.

    2016-01-01

    Objectives White spot lesions are the most undesired side-effect of fixed orthodontic treatments. The objectives of this study were to combine nanoparticles of silver (NAg) with 2-methacryloyloxyethyl phosphorylcholine (MPC) to develop a modified resin-modified glass ionomer cement (RMGI) as orthodontic cement with double benefits of antibacterial and protein-repellent capabilities for the first time. Methods NAg and MPC were incorporated into a commercial RMGI. Another commercial orthodontic adhesive also served as control. Enamel shear bond strengths (SBS) were determined. Protein adsorption was measured via a micro bicinchoninic acid method. A dental plaque microcosm biofilm model with human saliva as inoculum was tested. Biofilms adherent on the cement samples and planktonic bacteria in the culture medium away from the cement surfaces were both evaluated for bacterial metabolic activity, colony-forming units (CFU), and lactic acid production. Results Adding 0.1% NAg and 3% MPC to RMGI, and water-aging for 30 days, did not adversely affect the SBS, compared to the unmodified RMGI control (p>0.1). The modified RMGI containing 0.1% NAg and 3% MPC achieved the greatest reduction in protein adsorption, bacterial adhesion, CFU, metabolic activity and lactic acid production. The RMGI containing 0.1% NAg and 3% MPC inhibited not only the bacteria on its surface, but also the bacteria away from the surface in the culture medium. Conclusions The incorporation of double agents (antibacterial NAg + protein-repellent MPC) into RMGI achieved much stronger inhibition of biofilms than using each agent alone. The novel antibacterial and protein-repellent RMGI with substantially-reduced biofilm acids is promising as an orthodontic cement to combat white spot lesions in enamel. PMID:26427311

  15. Lightning activity in Saturn's Great White Spot of 2010/2011

    NASA Astrophysics Data System (ADS)

    Fischer, Georg; Dyudina, Ulyana A.; Sayanagi, Kunio M.; Pagaran, Joseph A.

    2014-05-01

    In this presentation we will summarize the main findings about Saturn's Great White Spot of 2010/2011 gained by analyzing data from the Cassini RPWS (Radio and Plasma Wave Science) instrument in combination with images from the Cassini cameras. Great White Spots (GWS) are planetary-scale disturbances in Saturn's atmosphere that usually happen once per Saturn year (29.5 Earth years). The last GWS of 2010/2011 occurred earlier than expected, and the Cassini RPWS instrument measured radio emissions caused by lightning discharges thereby identifying the GWS as a giant thunderstorm. Lightning radio emissions were measured for almost 9 months, from 5 December 2010 until 28 August 2011, with typical flash rates of more than 10 per second. Many images of the GWS were taken with the Cassini ISS (Imaging Science Subsystem) and by Earth-based telescopes which captured the storm's birth, evolution and demise. The GWS developed an elongated eastward tail due to Saturn's zonal winds, and this tail wrapped around the whole planet by February 2011. RPWS data indicated that the storm's head was the main center of lightning activity, but the region of active thunderstorm cells also extented eastward into the tail. This was confirmed by the first optical observation of lightning flashes on Saturn's dayside located eastward of the head. The head region periodically spawned anticyclonic vortices, and the optical flashes appeared in the cyclonic gaps between them where the atmosphere looked clear down to the level of deep clouds. The largest anticyclonic vortex in the tail drifted with a rate that was 2 deg/day slower than the head. Hence, after about half a year one caught up with the other, and it came to a head-vortex collision in mid-June 2011. This led to a significant decrease of lightning and convective activity, which became intermittent and finally ended in late August 2011.

  16. Virulence and genotypes of white spot syndrome virus infecting Pacific white shrimp Litopenaeus vannamei in north-western Mexico.

    PubMed

    Ramos-Paredes, J; Grijalva-Chon, J M; Ibarra-Gámez, J C

    2017-03-01

    White spot syndrome virus (WSSV) has caused substantial global economic impact on aquaculture, and it has been determined that strains can vary in virulence. In this study, the effect of viral load was evaluated by infecting Litopenaeus vannamei with 10-fold serial dilution of tissue infected with strain WSSV Mx-H, and the virulence of four WSSV strains from north-western Mexico was assessed along with their variable number of tandem repeat (VNTR) genotypes in ORF75, ORF94 and ORF125. The LD50 of the Mx-H strain was a dilution dose of 10(-7.5) ; the mortality titre was 10(9.2) LD50 per gram. In shrimp injected with 10(2.5) to 10(6.5) LD50 , no significant virulence differences were evident. Using mortality data, the four WSSV strains grouped into three virulence levels. The Mx-F strain (intermediate virulence) and the Mx-C strain (high virulence) showed more genetic differences than those observed between the Mx-G (low-virulence) and Mx-H (high-virulence) strains, in ORF94 and ORF125. The application of high-viral-load inocula proved useful in determining the different virulence phenotypes of the WSSV strains from the Eastern Pacific.

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

    PubMed

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

    2014-09-01

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

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

    PubMed

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

    2015-05-01

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

  19. Microphthalmia-associated transcription factor mutations are associated with white-spotted coat color in swamp buffalo.

    PubMed

    Yusnizar, Y; Wilbe, M; Herlino, A O; Sumantri, C; Noor, R Rachman; Boediono, A; Andersson, L; Andersson, G

    2015-12-01

    A candidate gene analysis of the microphthalmia-associated transcription factor (MITF) gene was used in an attempt to identify the genetic basis for a white-spotted coat color phenotype in the Asian swamp buffalo (Bubalus bubalis carabanensis). Ninety-three buffaloes-32 solid, 38 spotted and 23 white individuals-were Sanger-sequenced for all MITF exons as well as highly conserved intronic and flanking regions. MITF cDNA representing skin and iris tissue from six spotted, nine solid and one white buffaloes was also Sanger-sequenced to confirm detected mutations. Two independent loss-of-function mutations, a premature stop codon (c.328C>T, p.Arg110*) and a donor splice-site mutation (c.840+2T>A, p.Glu281_Leu282Ins8), both of which cause white-spotted coat color in swamp buffaloes, were identified. The nonsense mutation leads to a premature stop codon in exon 3, and likely removal of the resulting mRNA via nonsense-mediated decay pathway, whereas the donor splice-site mutation leads to aberrant splicing of exon 8 that encodes part of a highly conserved region of MITF. The resulting insertion of eight amino acid residues is expected to perturb the leucine zipper part in the basic helix-loop-helix leucine zipper (bHLH-Zip) domain and will most likely influence dimerization and DNA binding capacity. Electrophoretic mobility shift assay was performed using mutant and wild-type MITF proteins and showed that the mutant MITF protein resulting from the splice-site mutation decreased in vitro DNA binding capacity compared to wild-type MITF. White-spotted buffalo bulls are sacrificed in funeral ceremonies in Tana Toraja, Indonesia, because they are considered holy, and our results show that genetic variation causes a tie to the cultural use of these buffaloes.

  20. Effectiveness of hand sorting, flotation/washing, dehulling and combinations thereof on the decontamination of mycotoxin-contaminated white maize.

    PubMed

    Matumba, Limbikani; Van Poucke, Christof; Njumbe Ediage, Emmanuel; Jacobs, Bart; De Saeger, Sarah

    2015-01-01

    Maize is one of the major staple foods of Sub-Saharan Africa and is consumed as whole or dehulled grain. In this region, where the environmental conditions favour fungal growth and mycotoxin production, the majority of the population are subsistence consumers who, unfortunately, have little or no access to mycotoxin testing of their food. In an attempt to develop feasible reduction strategies in dietary mycotoxin exposure of the population, a three-factorial design experiment was conducted to examine and compare the efficacy of hand sorting, flotation, dehulling and combinations thereof in removing naturally occurring aflatoxins, fumonisins, nivalenol, deoxynivalenol and alternariol in shelled white maize. Regression analysis was used to determine the significant (p < 0.05) process variables on the removal of mycotoxins from the maize. Results from this experiment indicated that hand sorting had the greatest effect on mycotoxin removal, while flotation yielded the least effect. In particular hand sorting left < 6% of aflatoxin B1 and < 5% of fumonisin B1. Based on these results, hand sorting of maize grains is being recommended as a last line of defence against mycotoxin exposure among subsistence consumers.

  1. Changes in chemical composition and digestibility of three maize stover components digested by white-rot fungi.

    PubMed

    Lynch, J P; O'Kiely, P; Murphy, R; Doyle, E M

    2014-08-01

    Maize stover (total stem and leaves) is not considered a ruminant feed of high nutritive value. Therefore, an improvement in its digestibility may increase the viability of total forage maize production systems in marginal growth regions. The objective of this study was to describe the changes in chemical composition during the storage of contrasting components of maize stover (leaf, upper stem and lower stem) treated with either of two lignin degrading white-rot fungi (WRF; Pleurotus ostreatus, Trametes versicolor). Three components of maize stover (leaf, upper stem and lower stem), harvested at a conventional maturity for silage production, were digested with either of two WRF for one of four digestion durations (1-4 months). Samples taken prior to fungal inoculation were used to benchmark the changes that occurred. The degradation of acid detergent lignin was observed in all sample types digested with P. ostreatus; however, the loss of digestible substrate in all samples inoculated with P. ostreatus was high, and therefore, P. ostreatus-digested samples had a lower dry matter digestibility than samples prior to inoculation. Similarly, T. veriscolor-digested leaf underwent a non-selective degradation of the rumen-digestible components of fibre. The changes in chemical composition of leaf, upper stem and lower stem digested with either P. ostreatus or T. veriscolor were not beneficial to the feed value of the forage, and incurred high DM losses.

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

    PubMed

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

    2014-07-01

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

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

    PubMed Central

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

    2011-01-01

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

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

    PubMed

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

    2013-07-01

    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.

  5. Viral ubiquitin ligase WSSV222 is required for efficient white spot syndrome virus replication in shrimp.

    PubMed

    He, Fang; Syed, Syed Musthaq; Hameed, A S Sahul; Kwang, Jimmy

    2009-06-01

    The E3 ligase WSSV222 of white spot syndrome virus (WSSV) is involved in anti-apoptosis regulation by ubiquitin-mediated degradation of tumour suppressor-like protein (TSL), a shrimp tumour suppressor. In the present study, WSSV222 gene expression was silenced by using specific small interfering RNA (siRNA) in Sf9 and BHK cells. Based on the results of the in vitro silencing, WSSV-challenged shrimp were treated with anti-WSSV222 siRNA to knock down WSSV222 protein expression. The survival rate of shrimp and the efficiency of WSSV replication were assessed to evaluate the efficacy of anti-WSSV222 siRNA in regulating WSSV infection in shrimp. The anti-WSSV222 siRNA reduced the cumulative mortality in shrimp challenged with 10(3) copies of WSSV and delayed the mean time to death in shrimp challenged with the higher dose of 10(6) copies. The results of real-time quantitative PCR showed that virus replication was delayed and reduced in WSSV-challenged shrimp treated with anti-WSSV222 siRNA in comparison with challenged shrimp treated with random-control siRNA. Co-immunoprecipitation assays revealed that WSSV222 silencing inhibited the degradation of TSL in WSSV-challenged shrimp, indicating the requirement for WSSV222 for efficient replication of WSSV in shrimp.

  6. Laminin Receptor in Shrimp Is a Cellular Attachment Receptor for White Spot Syndrome Virus

    PubMed Central

    Liu, Wang-Jing; Li, Yi-Chieh; Kou, Guang-Hsiung; Lo, Chu-Fang

    2016-01-01

    White spot syndrome virus (WSSV, genus Whispovirus, family Nimaviridae) is causing huge economic losses in global shrimp farming, but there is no effective control. Shrimp cell laminin receptor (Lamr) may have a role in WSSV infection. The objective was to characterize interactions between Penaeus monodon Lamr (PmLamr) and WSSV structural proteins. In this study, PmLamr interacted with nine WSSV structural proteins (based on yeast two-hybrid screening), of which one (VP31) was characterized. Protein pull-down assay confirmed the interaction between PmLamr and VP31; the latter was an envelope protein exposed outside the WSSV virion (based on membrane topology assays). Furthermore, similar to mammalian Lamr, there were two major protein bands in shrimp cells. Cellular localization assay demonstrated VP31 co-localized with PmLamr on transfected cells. Enzyme-link immunosorbent assay (ELISA) and competitive ELISA demonstrated binding of VP31 on PmLamr was dose-dependent; however, addition of WSSV virion competed for binding affinity. Furthermore, based on an in vivo neutralization assay, both VP31 and PmLamr delayed mortality in shrimp challenged with WSSV. We concluded Lamr was an important receptor for WSSV infection and the viral envelope protein VP31 may have a role in host cell recognition and binding. These data contributed to elucidating pathogenesis of WSSV infection and may help in controlling this disease. PMID:27257954

  7. Laminin Receptor in Shrimp Is a Cellular Attachment Receptor for White Spot Syndrome Virus.

    PubMed

    Liu, Wang-Jing; Li, Yi-Chieh; Kou, Guang-Hsiung; Lo, Chu-Fang

    2016-01-01

    White spot syndrome virus (WSSV, genus Whispovirus, family Nimaviridae) is causing huge economic losses in global shrimp farming, but there is no effective control. Shrimp cell laminin receptor (Lamr) may have a role in WSSV infection. The objective was to characterize interactions between Penaeus monodon Lamr (PmLamr) and WSSV structural proteins. In this study, PmLamr interacted with nine WSSV structural proteins (based on yeast two-hybrid screening), of which one (VP31) was characterized. Protein pull-down assay confirmed the interaction between PmLamr and VP31; the latter was an envelope protein exposed outside the WSSV virion (based on membrane topology assays). Furthermore, similar to mammalian Lamr, there were two major protein bands in shrimp cells. Cellular localization assay demonstrated VP31 co-localized with PmLamr on transfected cells. Enzyme-link immunosorbent assay (ELISA) and competitive ELISA demonstrated binding of VP31 on PmLamr was dose-dependent; however, addition of WSSV virion competed for binding affinity. Furthermore, based on an in vivo neutralization assay, both VP31 and PmLamr delayed mortality in shrimp challenged with WSSV. We concluded Lamr was an important receptor for WSSV infection and the viral envelope protein VP31 may have a role in host cell recognition and binding. These data contributed to elucidating pathogenesis of WSSV infection and may help in controlling this disease.

  8. Crystal Structure of Major Envelope Protein VP24 from White Spot Syndrome Virus

    PubMed Central

    Sun, Lifang; Su, Yintao; Zhao, Yanhe; Fu, Zheng-qing; Wu, Yunkun

    2016-01-01

    White spot syndrome virus (WSSV) is one of the major and most serious pathogen in the shrimp industry. As one of the most abundant envelope protein, VP24 acts as a core protein interacting with other structure proteins and plays an important role in virus assembly and infection. Here, we have presented the crystal structure of VP24 from WSSV. In the structure, VP24 consists of a nine-stranded β–barrel fold with mostly antiparallel β-strands, and the loops extending out the β–barrel at both N-terminus and C-terminus, which is distinct to those of the other two major envelope proteins VP28 and VP26. Structural comparison of VP24 with VP26 and VP28 reveals opposite electrostatic surface potential properties of them. These structural differences could provide insight into their differential functional mechanisms and roles for virus assembly and infection. Moreover, the structure reveals a trimeric assembly, suggesting a likely natural conformation of VP24 in viral envelope. Therefore, in addition to confirming the evolutionary relationship among the three abundant envelope proteins of WSSV, our structural studies also facilitate a better understanding of the molecular mechanism underlying special roles of VP24 in WSSV assembly and infection. PMID:27572278

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

    PubMed Central

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

    2016-01-01

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

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

    PubMed

    Claydon, Kerry; Cullen, Bradford; Owens, Leigh

    2004-12-13

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

  11. Fusion of flagellin 2 with bivalent white spot syndrome virus vaccine increases survival in freshwater shrimp.

    PubMed

    Cho, Hansam; Park, Na Hye; Jang, Yuyeon; Gwon, Yong-Dae; Cho, Yeondong; Heo, Yoon-Ki; Park, Ki-Hoon; Lee, Hee-Jung; Choi, Tae Jin; Kim, Young Bong

    2017-03-01

    Despite large economic losses attributable to white spot syndrome virus (WSSV), an infectious pathogen of penaeid shrimp and other crustaceans worldwide, no efficient vaccines or antiviral agents to control the virus are available at present. Here, we designed and constructed baculovirus-based vaccines delivering genes encoding the WSSV envelope proteins, VP28 and VP19. To enhance the immunogenicity of the baculovirus-based vaccine, we fused a Salmonella typhimurium flagellin 2 (FL2) gene with VP28 or VP19 gene. Both vaccine constructs elicited similar high titlers of anti-WSSV IgG after oral immunization in mice. The protective effect of oral vaccines upon WSSV challenge was observed in Macrobrachium nipponense. Bivalent vaccine displaying WSSV envelope proteins, VP19 and VP28, led to enhanced more than 10% survival protection against WSSV infection, compared to monovalent vaccine containing WSSV envelope protein, VP19 or VP28. Furthermore, a baculovirus-based WSSV vaccine fused with FL2 gene, Ac-VP28-ie1VP19FL2, efficiently protected mice against WSSV challenge (89.5% survival rate). In support of the efficacy of FL2 in our vaccine, we verified FL2 enhanced survival rate and induced the NF-κB gene in Palaemon paucidens. The collective results strongly suggest that our recombinant baculoviral system displaying WSSV envelope protein and delivering FL2-fused WSSV envelope gene effectively induced protective responses, supporting the utility of a potential new oral DNA vaccine against WSSV.

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

    PubMed

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

    2013-11-01

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

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

    NASA Astrophysics Data System (ADS)

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

    2008-08-01

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

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

    PubMed

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

    2014-01-01

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

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

    PubMed

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

    2016-10-01

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

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

    PubMed

    Ye, Ting; Zong, Rongrong; Zhang, Xiaobo

    2012-08-01

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

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

    PubMed

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

    2014-01-01

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

  18. Envelope protein VP24 from White spot syndrome virus: expression, purification and crystallization.

    PubMed

    Sun, Lifang; Wu, Yunkun

    2016-08-01

    White spot syndrome virus (WSSV) is a major shrimp pathogen known to infect penaeid shrimp and other crustaceans. VP24 is one of the major envelope proteins of WSSV. In order to facilitate purification, crystallization and structure determination, the predicted N-terminal transmembrane region of approximately 26 amino acids was truncated from VP24 and several mutants were prepared to increase the proportion of selenomethionine (SeMet) residues for subsequent structural determination using the SAD method. Truncated VP24, its mutants and the corresponding SeMet-labelled proteins were purified, and the native and SeMet proteins were crystallized by the hanging-drop vapour-diffusion method. Crystals of VP24 were obtained using a reservoir consisting of 0.1 M Tris-HCl pH 8.5, 2.75 M ammonium acetate with a drop volume ratio of two parts protein solution to one part reservoir solution. Notably, ATP was added as a critical additive to the drop with a final concentration of 10 mM. Crystals of SeMet-labelled VP24 mutant diffracted to 3.0 Å resolution and those of the native diffracted to 2.4 Å resolution; the crystals belonged to space group I213, with unit-cell parameters a = b = c = 140 Å.

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

    PubMed

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

    2015-03-01

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

  20. Comparative genomic analysis of three white spot syndrome virus isolates of different virulence.

    PubMed

    Li, Fang; Gao, Meiling; Xu, Limei; Yang, Feng

    2017-04-01

    Three white spot syndrome virus (WSSV) isolates of different virulence were identified in our previous study, the high-virulent strain WSSV-CN01, the moderate-virulent strain WSSV-CN02 and the low-virulent strain WSSV-CN03. In this study, the genomes of these three WSSV isolates were sequenced, annotated and compared. The genome sizes for WSSV-CN01, WSSV-CN02, and WSSV-CN03 are 309,286, 294,261, and 284,148 bp, bearing 177, 164, and 154 putative protein-coding genes, respectively. The genomic variations including insertions, deletions, and substitutions were investigated. Thirty four genes show >20% variation in their sequences in WSSV-CN02 or WSSV-CN03, in comparison with WSSV-CN01, including six envelope protein genes (wsv237/vp41A, wsv238/vp52A, wsv338/vp62, wsv339/vp39, wsv077/vp36A, and wsv242/vp41B), and two immediate-early genes (wsv108 and wsv178). The genomic variations among WSSV isolates of different virulence, especially those in the coding regions, certainly provide new insight into the understanding of the molecular basis of WSSV pathogenesis.

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

    PubMed

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

    2014-05-01

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

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

    PubMed

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

    2015-07-01

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

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

    PubMed

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

    2015-09-01

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

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

    PubMed

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

    2013-10-01

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

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

    PubMed

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

    2012-12-03

    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.

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

    NASA Astrophysics Data System (ADS)

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

    2017-02-01

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

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

    PubMed

    Shekhar, M S; Ponniah, A G

    2015-07-01

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

  8. Construction and application of a protein interaction map for white spot syndrome virus (WSSV).

    PubMed

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

    2014-01-01

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

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

    PubMed

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

    2008-02-28

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

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

    PubMed Central

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

    2014-01-01

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

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

    NASA Technical Reports Server (NTRS)

    Dowling, Timothy E.; Ingersoll, Andrew P.

    1988-01-01

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

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

    PubMed

    Piamsomboon, Patharapol; Inchaisri, Chaidate; Wongtavatchai, Janenuj

    2015-12-09

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

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

    PubMed

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

    2015-01-01

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

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

    PubMed

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

    2012-02-01

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

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

    PubMed

    Caipang, Christopher Marlowe A; Aguana, Mary Paz N

    2010-10-01

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

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

    PubMed

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

    2009-04-01

    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.

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

    PubMed

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

    2013-04-01

    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.

  18. Scavenger Receptor C Mediates Phagocytosis of White Spot Syndrome Virus and Restricts Virus Proliferation in Shrimp.

    PubMed

    Yang, Ming-Chong; Shi, Xiu-Zhen; Yang, Hui-Ting; Sun, Jie-Jie; Xu, Ling; Wang, Xian-Wei; Zhao, Xiao-Fan; Wang, Jin-Xing

    2016-12-01

    Scavenger receptors are an important class of pattern recognition receptors that play several important roles in host defense against pathogens. The class C scavenger receptors (SRCs) have only been identified in a few invertebrates, and their role in the immune response against viruses is seldom studied. In this study, we firstly identified an SRC from kuruma shrimp, Marsupenaeus japonicus, designated MjSRC, which was significantly upregulated after white spot syndrome virus (WSSV) challenge at the mRNA and protein levels in hemocytes. The quantity of WSSV increased in shrimp after knockdown of MjSRC, compared with the controls. Furthermore, overexpression of MjSRC led to enhanced WSSV elimination via phagocytosis by hemocytes. Pull-down and co-immunoprecipitation assays demonstrated the interaction between MjSRC and the WSSV envelope protein. Electron microscopy observation indicated that the colloidal gold-labeled extracellular domain of MjSRC was located on the outer surface of WSSV. MjSRC formed a trimer and was internalized into the cytoplasm after WSSV challenge, and the internalization was strongly inhibited after knockdown of Mjβ-arrestin2. Further studies found that Mjβ-arrestin2 interacted with the intracellular domain of MjSRC and induced the internalization of WSSV in a clathrin-dependent manner. WSSV were co-localized with lysosomes in hemocytes and the WSSV quantity in shrimp increased after injection of lysosome inhibitor, chloroquine. Collectively, this study demonstrated that MjSRC recognized WSSV via its extracellular domain and invoked hemocyte phagocytosis to restrict WSSV systemic infection. This is the first study to report an SRC as a pattern recognition receptor promoting phagocytosis of a virus.

  19. Shrimp MyD88 responsive to bacteria and white spot syndrome virus.

    PubMed

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

    2013-02-01

    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.

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

    PubMed Central

    Hemerik, Lia; Vlak, Just M.

    2010-01-01

    Background White spot syndrome virus (WSSV) is the sole member of the novel Nimaviridae family, and the source of major economic problems in shrimp aquaculture. WSSV appears to have rapidly spread worldwide after the first reported outbreak in the early 1990s. Genomic deletions of various sizes occur at two loci in the WSSV genome, the ORF14/15 and ORF23/24 variable regions, and these have been used as molecular markers to study patterns of viral spread over space and time. We describe the dynamics underlying the process of WSSV genome shrinkage using empirical data and a simple mathematical model. Methodology/Principal Findings We genotyped new WSSV isolates from five Asian countries, and analyzed this information together with published data. Genome size appears to stabilize over time, and deletion size in the ORF23/24 variable region was significantly related to the time of the first WSSV outbreak in a particular country. Parameter estimates derived from fitting a simple mathematical model of genome shrinkage to the data support a geometric progression (k<1) of the genomic deletions, with k = 0.371±0.150. Conclusions/Significance The data suggest that the rate of genome shrinkage decreases over time before attenuating. Bioassay data provided support for a link between genome size and WSSV fitness in an aquaculture setting. Differences in genomic deletions between geographic WSSV isolates suggest that WSSV spread did not follow a smooth pattern of geographic radiation, suggesting spread of WSSV over long distances by commercial activities. We discuss two hypotheses for genome shrinkage, an adaptive and a neutral one. We argue in favor of the adaptive hypothesis, given that there is support for a link between WSSV genome size and fitness. PMID:20976239

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

    PubMed

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

    2015-06-15

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

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

    PubMed

    Syed Musthaq, Syed Khader; Kwang, Jimmy

    2014-10-01

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

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

    SciTech Connect

    Tang,X.; Hew, C.

    2007-01-01

    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.

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

    PubMed

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

    2016-07-01

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

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

    PubMed

    Syed Musthaq, Syed Khader; Kwang, Jimmy

    2015-02-01

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

  6. VP24 Is a Chitin-Binding Protein Involved in White Spot Syndrome Virus Infection

    PubMed Central

    Li, Zaipeng; Han, Yali; Xu, Limei

    2015-01-01

    ABSTRACT Oral ingestion is the major route of infection for the white spot syndrome virus (WSSV). However, the mechanism by which virus particles in the digestive tract invade host cells is unknown. In the present study, we demonstrate that WSSV virions can bind to chitin through one of the major envelope proteins (VP24). Mutagenesis analysis indicated that amino acids (aa) 186 to 200 in the C terminus of VP24 were required for chitin binding. Moreover, the P-VP24186–200 peptide derived from the VP24 chitin binding region significantly inhibited the VP24-chitin interaction and the WSSV-chitin interaction, implying that VP24 participates in WSSV binding to chitin. Oral inoculation experiments showed that P-VP24186–200 treatment reduced the number of virus particles remaining in the digestive tract during the early stage of infection and greatly hindered WSSV proliferation in shrimp. These data indicate that binding of WSSV to chitin through the viral envelope protein VP24 is essential for WSSV per os infection and provide new ideas for preventing WSSV infection in shrimp farms. IMPORTANCE In this study, we show that WSSV can bind to chitin through the envelope protein VP24. The chitin-binding domain of VP24 maps to amino acids 186 to 200 in the C terminus. Binding of WSSV to chitin through the viral envelope protein VP24 is essential for WSSV per os infection. These findings not only extend our knowledge of WSSV infection but also provide new insights into strategies to prevent WSSV infection in shrimp farms. PMID:26512091

  7. Scavenger Receptor C Mediates Phagocytosis of White Spot Syndrome Virus and Restricts Virus Proliferation in Shrimp

    PubMed Central

    Yang, Ming-Chong; Shi, Xiu-Zhen; Yang, Hui-Ting; Sun, Jie-Jie; Xu, Ling; Wang, Xian-Wei; Zhao, Xiao-Fan

    2016-01-01

    Scavenger receptors are an important class of pattern recognition receptors that play several important roles in host defense against pathogens. The class C scavenger receptors (SRCs) have only been identified in a few invertebrates, and their role in the immune response against viruses is seldom studied. In this study, we firstly identified an SRC from kuruma shrimp, Marsupenaeus japonicus, designated MjSRC, which was significantly upregulated after white spot syndrome virus (WSSV) challenge at the mRNA and protein levels in hemocytes. The quantity of WSSV increased in shrimp after knockdown of MjSRC, compared with the controls. Furthermore, overexpression of MjSRC led to enhanced WSSV elimination via phagocytosis by hemocytes. Pull-down and co-immunoprecipitation assays demonstrated the interaction between MjSRC and the WSSV envelope protein. Electron microscopy observation indicated that the colloidal gold-labeled extracellular domain of MjSRC was located on the outer surface of WSSV. MjSRC formed a trimer and was internalized into the cytoplasm after WSSV challenge, and the internalization was strongly inhibited after knockdown of Mjβ-arrestin2. Further studies found that Mjβ-arrestin2 interacted with the intracellular domain of MjSRC and induced the internalization of WSSV in a clathrin-dependent manner. WSSV were co-localized with lysosomes in hemocytes and the WSSV quantity in shrimp increased after injection of lysosome inhibitor, chloroquine. Collectively, this study demonstrated that MjSRC recognized WSSV via its extracellular domain and invoked hemocyte phagocytosis to restrict WSSV systemic infection. This is the first study to report an SRC as a pattern recognition receptor promoting phagocytosis of a virus. PMID:28027319

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

    PubMed

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

    2016-05-01

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

  9. Environmental factors related to fungal infection and fumonisin accumulation during the development and drying of white maize kernels.

    PubMed

    Cao, Ana; Santiago, Rogelio; Ramos, Antonio J; Marín, Sonia; Reid, Lana M; Butrón, Ana

    2013-06-03

    In Southern Europe where whole maize kernels are ground and used for making bread and other food products, infection of the kernels by Fusarium verticillioides and subsequent fumonisin contamination pose a serious safety issue. The influence of environmental factors on this fungal infection and mycotoxin accumulation as the kernel develops has not been fully determined, especially in such food grade maize. The objectives of the present study were to determine which environmental factors may contribute to kernel invasion by F. verticillioides and fumonisin accumulation as kernels develop and dry in naturally infected white maize. Three maize hybrids were planted at two different sowing dates and kernel samples were collected 20, 40, 60, 80 and 100 days after silking. The percentage of kernels infected, and ergosterol and fumonisin contents were recorded for each sampling. F. verticillioides was the most prevalent species identified as the kernels developed. Temperature and moisture conditions during the first 80 days after silking favored natural kernel infection by F. verticillioides rather than by Aspergillus or Penicillium species. Fumonisin was found in kernels as early as 20 days after silking however significant fumonisin accumulation above levels acceptable in the EU did not occur until after physiological maturity of the kernel indicating that kernel drying in the field poses a high risk. Our results suggest that this could be due to increasing kernel damage by insects that favor fungal development, such as the damage by the moth Sitotroga cerealella, and to the occurrence of stress conditions for F. verticillioides growth that could trigger fumonisin biosynthesis, such as exposure to suboptimal temperatures for growth simultaneously with low water activity.

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

    PubMed Central

    Gilbert, Kenneth; Joseph, Raphael; Vo, Alex; Patel, Trusha; Chaudhry, Samiya; Nguyen, Uyen; Trevor, Amy; Robinson, Erica; Campbell, Margaret; McLennan, John; Houran, Farielle; Wong, Tristan; Flann, Kendra; Wages, Melissa; Palmer, Elizabeth A.; Peterson, John; Engle, John; Maier, Tom; Machida, Curtis A.

    2014-01-01

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

  11. Minichromosome maintenance protein 7 regulates phagocytosis in kuruma shrimp Marsupenaeus japonicas against white spot syndrome virus.

    PubMed

    Wang, Zhi; Zhu, Fei

    2016-08-01

    Minichromosome maintenance protein (MCM7) belongs to the MCM protein family and participates in the MCM complex by playing a role in the cell replication cycle and chromosome initiation of eukaryotes. Previously, we found that several genes, including MCM7, were over-expressed in Drosophila melanogaster after white spot syndrome virus (WSSV) infection. In this study, we aimed to further research the MCM7 of kuruma shrimp, Marsupenaeus japonicus (mjMCM7) and determine its role in the innate immune system. To this end, we cloned the entire 2307-bp mjMCM7 sequence, including a 1974-bp open reading frame (ORF) encoding a 658-aa-long protein. Real-time PCR showed that the gene was primarily expressed in the hemolymph and hepatopancreas and over-expressed in shrimp challenged with WSSV. Gene function study was carried out by knocking down the expression of MCM7 using small interference RNA (siRNA). The results revealed that β-actin, hemocyanin, prophenoloxidase (proPO) and tumor necrosis factor-α (TNF-α) were up-regulated while the cytoskeleton proteins such as myosin and Rho were significantly down-regulated at 24 h after treatment. The results indicate a possible relationship between mjMCM7 and the innate immune system, and suggest that mjMCM7 may play a role in phagocytosis. After WSSV challenge, WSSV copies and mortality count were both higher in the MCM7-siRNA-treated groups at 60 h after treatment, and the mortality count approached that of the control groups over time. The phagocytosis rate was significantly lower in the MCM7-siRNA-treated group than in the WSSV group. The findings of this study confirm that mjMCM7 positively regulates phagocytosis and plays an important role against WSSV. These results could help researchers to further understand the function of the MCM7 protein and reveal its potential role in the innate immunity of invertebrates.

  12. Temporal Evolution of Saturn's Great White Spot Storm 2010-2011

    NASA Astrophysics Data System (ADS)

    Sayanagi, K. M.; Ewald, S. P.; Dyudina, U. A.; Ingersoll, A. P.; Porco, C.; Muro, G. S.

    2011-12-01

    We report on the temporal evolution of a large cumulus storm that erupted on Saturn in early December of 2010. The new event marks the latest occurrence of the 30-year quasi-periodic giant storms on Saturn known as the Great White Spots (GWS), which last erupted in 1990. Cassini ISS camera first captured the storm on December 5th, 2010, and has since monitored the storm at a semi-regular interval. The current storm erupted at 33 degree N planetocentric latitude, and our measurements indicate that the storm's initial longitude coincided that of a feature called the String of Pearls (SoPs) first found using Cassini VIMS (Momary et al. 2006 DPS/AAS, Choi et al. 2009 Icarus 10.1029/2008JE003254), suggesting that the new GWS and SoPs may be causally related. Our ISS images between December 2010 and June 2011 show that the storm had a very bright leading edge, which drifted westward relative to the Voyager longitude system at 2.79 degree per Earth day, similar to that measured by Sanchez-Lavega et al (2011, Nature 10.1038/nature10203). Our new methane filter images (MT2 and MT3) show that a substantial amount of cloud material is lifted at the leading edge up to the stratosphere, which indicates highly energetic cumulus convective activities consistent with Fischer et al (2011, Nature 10.1038/nature10205)'s detection of lightning discharges in the radio frequencies. Our images also reveal vertical shear in the local wind fields around the storm. During the early phase of the storm a large dark oval formed near the leading edge of the storm and drifted westward at 0.85 degree per day on average between December 2010 and June 2011, which was also noted in CIRS observation (Fletcher et al. 2011, Science 10.1126/science.1204774). Our measurements show that the oval has an anticyclonic vorticity. The anticyclonic oval defined the eastern end of the storm activities. By late June 2011, the storm's leading edge collided with the anticyclonic oval after encircling the entire

  13. Transposon elements in maize white seedling 3, allele w3-8686, w3-kermicle-1 and w3-88-89-3563-33

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Maize white seedling 3 (w3) has served as a model albino-seedling mutant since its discovery in 1923. We show that the w3 phenotype is caused by disruptions in homogentisate solanesyl transferase (HST), an enzyme that catalyzes the committed step in plastoquinone-9 (PQ9) biosynthesis. This reaction ...

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

    PubMed

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

    2012-07-01

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

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

    PubMed

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

    2014-10-01

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

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

    PubMed

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

    2015-05-11

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

  17. Increasing production in Korean shrimp farms with white-spot syndrome virus PCR-negative brood stock.

    PubMed

    Seok, Hyeok Seung; Baek, Min Won; Lee, Hui Young; Kim, Dong Jae; Chun, Myung Sun; Kim, Jong Sheek; Chang, Se Ok; Park, Jae Hak

    2007-03-01

    White-spot syndrome virus (WSSV) is a devastating, infectious virus affecting shrimp. Although sensitive techniques involving PCR have been developed to assist farmers in screening shrimp (brood stock) for WSSV prior to stocking ponds, such practices have not yet been applied in Korea. Despite the rationality of implementing screening, there has been some doubt as to whether the stocking of WSSV-PCR-negative fry epidemiologically decreases white-spot disease outbreaks. Here, we report a retrospective analysis of data from shrimp farms in the western coast of Korea where WSSV-PCR-negative brood stocks were used to stock rearing ponds. A total of 366 shrimp from Heuksan Island were sampled for WSSV with PCR. Of the tested shrimp, 7.2% (28 brood stocks) were identified as WSSV positive; only WSSV-PCR-negative shrimp were used for brood stocks. Total unit production (final shrimp production/ the area of the ponds) was higher, at 1.96, in ponds where WSSV-PCR-negative shrimp were used, as compared with 1.02 in other ponds in Korea in 2004. This retrospective analysis of WSSV in Korea may be useful to the shrimp aquaculture industry, suggesting a testable hypothesis that may contribute to the eventual control of WSSV outbreaks.

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

    PubMed

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

    2014-09-01

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

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

    PubMed

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

    2012-09-15

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

  20. Microscopic analysis of "iron spot" on blue-and-white porcelain from Jingdezhen imperial kiln in early Ming dynasty (14th-15th century).

    PubMed

    Wang, Wenxuan; Zhu, Jian; Jiang, Jianxin; Xu, Changqing; Wu, Shurong; Guan, Li; Zhang, Zhaoxia; Wu, Menglei; Du, Jingnan

    2016-11-01

    "Sumali," as an imported cobalt ore from overseas, was a sort of precious and valuable pigment used for imperial kilns only, which produces characteristic "iron spot" to blue-and-white porcelain in early Ming Dynasty (A.D. 14th-15th century). Although there were some old studies on it, the morphology and formation of iron spot has not been fully investigated and understood. Therefore, five selected samples with typical spot from Jingdezhen imperial kiln in Ming Yongle periods (A.D. 1403-1424) were analyzed by various microscopic analysis including 3D digital microscope, SEM-EDS and EPMA. According to SEM images, samples can be divided into three groups: un-reflected "iron spot" without crystals, un-reflected "iron spot" with crystals and reflected "iron spot" with crystals. Furthermore, 3D micro-images revealed that "iron spots" separate out dendritic or snow-shaped crystals of iron only on and parallel to the surface of glaze for which "iron spot" show strong metallic luster. Combining with microscopic observation and microanalysis on crystallization and non-crystallization areas, it indicates that firing oxygen concentration is the ultimate causation of forming reflective iron spot which has a shallower distribution below the surface and limits crystals growing down. More details about characters of "iron spot" used "Sumali" were found and provided new clues to coloration, formation mechanism and porcelain producing technology of imperial kiln from 14th to 15th centuries of China.

  1. Lipophilic metabolite profiling of maize and sorghum seeds and seedlings, and their pest spotted stem borer larvae: a standardized GC-MS based approach.

    PubMed

    Kumar, Sandeep; Dhillon, Mukesh K

    2015-03-01

    In order to better understand the biochemical interactions and to identify new biomarkers for plant resistance against insects, we proposed a suitable lipophilic profiling method for insects and their host plants. The critical components of GC-MS based analysis are: sample amount, extraction, derivatization, temperature gradient, run time, and identification of peaks. For lipophilic metabolite profiling of maize and sorghum, and their insect pest, spotted stem borer larvae, we recommend 100 mg sample weight for seeds and insect samples (whole insect body), and 200 mg for seedlings. Maize and sorghum seeds required less time for fat extraction in comparison to their seedlings and the pest fed on these seedlings. GC-MS was standardized for better separation and intensity of peaks using different temperature gradients in the range of 180-300 C. A total of 48 lipophilic compounds encompassing various classes based on their functional groups such as fatty acids, fatty alcohols, hydrocarbons, sterols and terpenoids, vitamin derivative, etc. were separated in the seedlings (30), seeds (14), and the pest (26) in the retention time range of 3.22 to 29.41 min. This method could be useful to study nutritional aspects of different field crops in relation to various stresses apart from the analysis of lipophilic compounds for better understanding of insect-plant interactions.

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

    PubMed

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

    2014-08-21

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

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

    NASA Astrophysics Data System (ADS)

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

    2016-04-01

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

  4. The effects of nitrogen form on root morphological and physiological adaptations of maize, white lupin and faba bean under phosphorus deficiency

    PubMed Central

    Liu, Haitao; Tang, Caixian; Li, Chunjian

    2016-01-01

    Root morphological/physiological modifications are important for phosphorus (P) acquisition of plants under P deficiency, but strategies differ among plant species. Detailed studies on the response of maize roots to P deficiency are limited. Nitrogen (N) form influences root morphology/physiology, and thus may influence root responses to P deficiency. This work investigated adaptive mechanisms of maize roots to low P by comparison with white lupin and faba bean supplied with two N forms. Plants were grown for 7–16 days in hydroponics with sufficient (250 µmol L−1) and deficient P supply (1 µmol L−1) under supply of NH4NO3 or Ca(NO3)2. Plant growth and P uptake were measured, and release of protons and organic acid anions, and acid phosphatase activity in the root were monitored. The results showed that P deficiency significantly decreased shoot growth while increased root growth and total root length of maize and faba bean, but not white lupin. It enhanced the release of protons and organic acid anions, and acid phosphatase activity, from the roots of both legumes but not maize. Compared with Ca(NO3)2, NH4NO3 dramatically increased proton release by roots but did not alter root morphology or physiology of the three species in response to low P. It is concluded that the N form did not fundamentally change root morphological/physiological responses of the three species to P deficiency. Morphological variation in maize and morpho-physiological modifications in white lupin and faba bean were the main adaptive strategies to P deficiency. PMID:27519912

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

    PubMed

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

    2014-10-01

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

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

    PubMed

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

    2015-07-01

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

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

    PubMed

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

    2015-05-11

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

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

    PubMed

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

    2015-03-16

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

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

    SciTech Connect

    Li Li; Xie Xixian; Yang Feng . E-mail: mbiotech@public.xm.fj.cn

    2005-09-15

    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.

  10. Two White Spot Syndrome Virus MicroRNAs Target the Dorsal Gene To Promote Virus Infection in Marsupenaeus japonicus Shrimp.

    PubMed

    Ren, Qian; Huang, Xin; Cui, Yalei; Sun, Jiejie; Wang, Wen; Zhang, Xiaobo

    2017-04-15

    In eukaryotes, microRNAs (miRNAs) serve as regulators of many biological processes, including virus infection. An miRNA can generally target diverse genes during virus-host interactions. However, the regulation of gene expression by multiple miRNAs has not yet been extensively explored during virus infection. This study found that the Spaztle (Spz)-Toll-Dorsal-antilipopolysaccharide factor (ALF) signaling pathway plays a very important role in antiviral immunity against invasion of white spot syndrome virus (WSSV) in shrimp (Marsupenaeus japonicus). Dorsal, the central gene in the Toll pathway, was targeted by two viral miRNAs (WSSV-miR-N13 and WSSV-miR-N23) during WSSV infection. The regulation of Dorsal expression by viral miRNAs suppressed the Spz-Toll-Dorsal-ALF signaling pathway in shrimp in vivo, leading to virus infection. Our study contributes novel insights into the viral miRNA-mediated Toll signaling pathway during the virus-host interaction.IMPORTANCE An miRNA can target diverse genes during virus-host interactions. However, the regulation of gene expression by multiple miRNAs during virus infection has not yet been extensively explored. The results of this study indicated that the shrimp Dorsal gene, the central gene in the Toll pathway, was targeted by two viral miRNAs during infection with white spot syndrome virus. Regulation of Dorsal expression by viral miRNAs suppressed the Spz-Toll-Dorsal-ALF signaling pathway in shrimp in vivo, leading to virus infection. Our study provides new insight into the viral miRNA-mediated Toll signaling pathway in virus-host interactions.

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

    SciTech Connect

    Syed Musthaq, S.; Madhan, Selvaraj; Sahul Hameed, A.S.; Kwang, Jimmy

    2009-09-01

    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.

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

    PubMed

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

    2012-09-01

    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 μL/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.

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

    PubMed

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

    2015-07-01

    Immunity parameters and the expression levels of several immune-related proteins, including lipopolysaccharide and β-glucan binding protein (LGBP), peroxinectin (PX), intergin β (IB), prophenoloxidase (proPO) I, proPO II, α2-macroglobulin (α2-M), cytosolic mangangese superoxide dismutase (cytMnSOD), mitochondria manganese superoxide dismutase (mtMnSOD), catalase, glutathione peroxidase (GPx), lysozyme, and penaeidin 3a were examined in white shrimp Litopenaeus vannamei reared at stocking densities of 2, 10, 20, 30, and 40 shrimp L(-1) after 3, 6, and 12 h. All immune parameters including haemocyte count, phenoloxidase (PO) activity, respiratory burst (RB), superoxide dismutase (SOD) activity, lysozyme activity, and haemolymph protein were negatively related to density and time. The PO activity, SOD activity, and lysozyme activity of shrimp reared at 10 shrimp L(-1) after 12 h significantly decreased. The transcript levels of these immune-related proteins were down-regulated in shrimp reared at 20, 30, and 40 shrimp L(-1) after 12 h. Phagocytic activity and clearance efficiency to Vibrio alginolyticus were significantly lower in shrimp reared at 30 and 40 shrimp L(-1) after 12 h. The mortality rates of shrimp reared at 20 and 40 shrimp L(-1) were significantly higher than shrimp reared at 2 shrimp L(-1) over 12-144 h and 12-48 h, respectively. Shrimp reared at high densities (>10 shrimp L(-1)) exhibited decreased resistance against pathogens as evidenced by reductions in immune parameters together with decreased expression levels of immune-related proteins, indicating perturbations of the immune system.

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

    PubMed

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

    2011-12-01

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

  15. Expressing the maize anthocyanin regulatory gene Lc increased flavonoid content in the seed of white pericarp rice and purple pericarp rice.

    PubMed

    Song, Y E; Wang, X; Shen, Z W; Xu, Y; Li, J Y

    2013-11-01

    The colour of red, purple, brown and white occurs in pericarp of rice. Here, the maize anthocyanin regulatory gene Lc under control of the promoter of the rice glutelin gene Gt1 was introduced in the white pericarp rice "Chao2-10" and purple pericarp rice "Qingjiaozidao". The results demonstrated that some transgenic "Chao2-10" rice pericarps became brown, and the total flavonoid contents in the unpolished rice of the two transgenic rices increased significantly compared with their respective controls. Unpolished rice kernel thickness and weight in the two transgenic rices decreased slightly.

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

    PubMed

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

    2014-02-01

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

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

    PubMed

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

    2011-01-01

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

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

    PubMed

    Mishra, S S; Shekhar, M S

    2005-07-01

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

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

    NASA Astrophysics Data System (ADS)

    Baines, Kevin H.; Carlson, Robert W.

    1999-01-01

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

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

    PubMed

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

    2011-12-01

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

  1. Gene expression profiling in gill tissues of White spot syndrome virus infected black tiger shrimp Penaeus monodon by DNA microarray.

    PubMed

    Shekhar, M S; Gomathi, A; Gopikrishna, G; Ponniah, A G

    2015-06-01

    White spot syndrome virus (WSSV) continues to be the most devastating viral pathogen infecting penaeid shrimp the world over. The genome of WSSV has been deciphered and characterized from three geographical isolates and significant progress has been made in developing various molecular diagnostic methods to detect the virus. However, the information on host immune gene response to WSSV pathogenesis is limited. Microarray analysis was carried out as an approach to analyse the gene expression in black tiger shrimp Penaeus monodon in response to WSSV infection. Gill tissues collected from the WSSV infected shrimp at 6, 24, 48 h and moribund stage were analysed for differential gene expression. Shrimp cDNAs of 40,059 unique sequences were considered for designing the microarray chip. The Cy3-labeled cRNA derived from healthy and WSSV-infected shrimp was subjected to hybridization with all the DNA spots in the microarray which revealed 8,633 and 11,147 as up- and down-regulated genes respectively at different time intervals post infection. The altered expression of these numerous genes represented diverse functions such as immune response, osmoregulation, apoptosis, nucleic acid binding, energy and metabolism, signal transduction, stress response and molting. The changes in gene expression profiles observed by microarray analysis provides molecular insights and framework of genes which are up- and down-regulated at different time intervals during WSSV infection in shrimp. The microarray data was validated by Real Time analysis of four differentially expressed genes involved in apoptosis (translationally controlled tumor protein, inhibitor of apoptosis protein, ubiquitin conjugated enzyme E2 and caspase) for gene expression levels. The role of apoptosis related genes in WSSV infected shrimp is discussed herein.

  2. Cytosolic manganese superoxide dismutase genes from the white shrimp Litopenaeus vannamei are differentially expressed in response to lipopolysaccharides, white spot virus and during ontogeny.

    PubMed

    Gómez-Anduro, Gracia A; Ascencio-Valle, Felipe; Peregrino-Uriarte, Alma Beatriz; Cámpa-Córdova, Angel; Yepiz-Plascencia, Gloria

    2012-08-01

    Manganese superoxide dismutase (MnSOD) is an antioxidant enzyme usually located in mitochondria. There are only a few examples of cytosolic MnSOD (cMnSOD). In the shrimp Litopenaeus vannamei, we have previously characterized three cMnSOD cDNAs and their differential tissue-specific expression. To obtain insights about their genomic organization, we characterized the three corresponding cMnSOD genes, named them cMnsod1, cMnsod2, and cMnsod3 and studied their specific expression during ontogeny, response to lipopolysaccharides (LPS) and white spot virus infection (WSSV) in hemocytes from shrimp. The first two genes contain five introns flanked by canonical 5'-GT-AG-3' intron splice-site junctions, while the third one is intron-less. We analyzed 995 nucleotides upstream cMnsod2, but no classical promoter sequences were found. The deduced products of the three cMnSOD genes differ in two amino acids and there are four silent changes. cMnsod3 expression is modulated by WSSV and cMnsod2 by LPS. cMnsod2 is expressed from eggs to post larval stage during ontogeny. This is the first report of crustacean cMnSOD multigenes that are differently induced during the defense response and ontogeny.

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

    NASA Astrophysics Data System (ADS)

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

    2004-10-01

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

  4. Ferritin administration effectively enhances immunity, physiological responses, and survival of Pacific white shrimp (Litopenaeus vannamei) challenged with white spot syndrome virus.

    PubMed

    Ruan, Yuan-Hwa; Kuo, Ching-Ming; Lo, Chu-Fang; Lee, Min-Hsien; Lian, Juang-Lin; Hsieh, Shu-Ling

    2010-04-01

    We examined the physiological (hemolymph glucose, lactate, and lipid) and innate non-specific immune responses (total hemocyte count (THC), phenoloxidase (PO) activity, respiratory bursts (release of superoxide anion, O(2)(-)) and superoxide dismutase (SOD) activity) to white spot syndrome virus (WSSV) in white shrimp (Litopenaeus vannamei) that were individually injected with 0.1, 0.5, and 1 ng g(-1) ferritin. Results showed that the THC, PO activity, and respiratory bursts of L. vannamei obviously increased (p < 0.05) 12 h after being injected with any dose of ferritin. However, the THC, PO activity, and respiratory bursts of L. vannamei that had received 0.5 and 1 ng g(-1) ferritin were significant higher than those of the other groups at 36-60, 60-72, and 36-60 h, respectively. SOD activities of L. vannamei 12 h after receiving 0.1, 0.5, and 1 ng g(-1) ferritin were significantly higher than those receiving saline. L. vannamei injected with ferritin at any dose maintained lower glucose, lactate, and lipid levels in response to WSSV challenge after 12-36, 24-48, and 36-60 h, respectively. The survival of shrimp that had received 0.5 and 1 ng g(-1) ferritin was significantly higher than that of shrimp that received saline and of control shrimp after 72 h. The ferritin messenger RNA transcripts of shrimp that had received 0.5 and 1 ng g(-1) ferritin were significantly higher than that of shrimp that received saline after 36 h. It was, therefore, concluded that the immune ability and resistance against WSSV infection increased in L. vannamei that had received > 0.5 ng g(-1) ferritin. Ferritin does play important roles in the innate immunity of the white shrimp. We observed higher SOD activities of L. vannamei that had received 0.1, 0.5, and 1 ng ferritin after 12 h than those that had received only saline (control), and the high SOD expression remained at the same levels even after 72 h of treatment.

  5. Risk factors associated with white spot syndrome virus infection in a Vietnamese rice-shrimp farming system.

    PubMed

    Corsin, F; Turnbull, J F; Hao, N V; Mohan, C V; Phi, T T; Phuoc, L H; Tinh, N T; Morgan, K L

    2001-10-29

    White spot disease (WSD) is a pandemic disease caused by a virus commonly known as white spot syndrome virus (WSSV). Several risk factors for WSD outbreaks have been suggested. However, there have been very few studies to identify risk factors for WSD outbreaks in culture systems. This paper presents and discusses the risk factors for WSSV infection identified during a longitudinal observational study conducted in a Vietnamese rice-shrimp farming system. A total of 158 variables were measured comprising location, features of the pond, management practices, pond bottom quality, shrimp health and other animals in the pond. At the end of the study period WSSV was detected in 15 of the 24 ponds followed through the production cycle (62.5%). One hundred and thirty-nine variables were used in univariate analyses. All the variables with a p-value < or = 0.10 were used in unconditional logistic regression in a forward stepwise model. An effect of location was identified in both univariate and multivariate analyses showing that ponds located in the eastern portion of the study site, closer to the sea, were more likely to test positive for WSSV by 1-step PCR at harvest. Ponds with shrimp of a smaller average size 1 mo after stocking tended to be positive for WSSV at the end of the production cycle. Average weight at 1 mo was also highlighted in multivariate analyses when considered as either a risk factor or an outcome. Other risk factors identified in univariate analyses were earlier date of stocking and use of commercial feed. A number of variables also appeared to be associated with a reduced risk of WSSV at harvest including the presence of dead post larvae in the batch sampled at stocking, presence of Hemigrapsus spp. crabs during the first month of production, feeding vitamin premix or legumes, presence of high numbers of shrimp with bacterial infection and the presence of larger mud crabs or gobies at harvest. No associations were detected with WSSV at harvest and

  6. Differential gene expression profile of the hepatopancreas of white spot syndrome virus infected Fenneropenaeus indicus by suppression subtractive hybridization.

    PubMed

    James, Roswin; Thampuran, Nirmala; Lalitha, K V; Rajan, Lawrance Anbu; Joseph, Toms C

    2010-11-01

    Suppression Subtractive Hybridization was employed in order to identify the differentially expressed genes in the hepatopancreas of white spot syndrome virus infected Fenneropenaeus indicus. A forward subtracted cDNA library generated 356 clones following a white spot syndrome virus infection. A total of 345 clones with more than 100 nucleotides were selected for further analysis using bioinformatics tools after vector screening. Twenty-three contigs and 111 singletons were generated from a total of 134 consensuses. The consensuses, on a sequence homology search using BLASTX (NCBI), revealed that 74 (55%) of them had no significant match to reported sequences in the database, suggesting that they were found for the first time and are probably associated with shrimp immune function. Out of the remaining 60 (45%) consensuses, 43 had significant homology to known protein sequences in the database while 17 consensuses are homologous to unknown proteins in the database which are considered novel. The most abundant genes in the subtracted library were antimicrobial peptides accounting for 56 clones; among which one is a member of SNF2 family of proteins and another belonged to PfP1 family of proteins on analysis using Antimicrobial peptide predictor software. The other predicted genes in the subtracted library include signal transduction molecules (GTPase, Serine threonine kinase, Armadillo repeats etc), antioxidant enzymes (Cytochrome oxidase, Monomeric sarcosine oxidase and Catalase), active transporters (Nuclear Localization Signal [NLS], calcium ATPase, sodium glutamate symporter, Store-Operated Calcium Entry [SOCE] and ribonucleoprotein [RNP]) contributing to 19, 14 and 5 clones respectively. Three clones are homologous to reverse transcriptase; a first time report in shrimp and one each belong to cell adhesion molecule and Proteinase. InterProScan at EMBL, when used for an integrated search at PROSITE predicted; signal sequences and transmembrane regions for 13

  7. Identification and analysis of host plant resistance in leading maize genotypes against spotted stem borer, Chilo partellus (Swinhoe) (Lepidoptera: Pyralidae).

    PubMed

    Arabjafari, K H; Jalali, S K

    2007-06-01

    Twenty six popular varieties of maize in Karnataka state (India) were screened for resistance to Chilo partellus (Swinhoe). In field trial, varieties CM132, CM137 and PMZ103 showed the highest level of resistance, respectively. While the varieties GK3014, HY4642 and DK984 had the lowest level of resistance, respectively. In glasshouse trial, the varieties CM137 and HY4642 had the highest and lowest level of resistance. A positive correlation equal to +0.111 (in field) and +0.521 (in glasshouse) was found between the Leaf Injury Score (LIS) and Stem Diameter (SD). Meanwhile, negative correlation equal to -0.432 (in field) and -0.896 (in glasshouse) was found between Leaf Injury Score (LIS) and Internodal Distance (InD). Results of the studies on major nutritional elements on resistant variety CM137 and susceptible variety HY4642 indicated higher percentage of P, K, Fe and Si in stem tissues of the variety CM137, while the variety HY4642 had higher percentage of N and Sugar, highlighting the role of these elements in conferring resistance and susceptibility to C. partellus. Studies on the impact of resistance factors on biological events of C. partellus revealed significant differences between percentage survival of larvae, percentage of pupation, pupal weight and pupal period (females only) on two varieties CM137 and HY4642. But there was no significant difference in the percentage of moth emergence on two varieties.

  8. Critical evaluation of incidence and prevalence of white spot lesions during fixed orthodontic appliance treatment: A meta-analysis

    PubMed Central

    Sundararaj, Dhinahar; Venkatachalapathy, Sudhakar; Tandon, Akshay; Pereira, Aaron

    2015-01-01

    Objective: Development of dental caries, specifically, white spot lesions (WSLs), continues to be a well-recognized and troubling side effect of orthodontic fixed appliance therapy, despite vast improvement in preventive dental techniques and procedures. The aim of this meta-analysis is to evaluate, determine, and summarize the incidence and prevalence rates of WSLs during orthodontic treatment that have been published in the literature. Materials and Methods: According to predetermined criteria, databases were searched for appropriate studies. References of the selected articles and relevant reviews were searched for any missed publications. Results: In the 14 studies evaluated for WSLs, the incidence of new carious lesions formed during orthodontic treatment in patients was 45.8% and the prevalence of lesions in patients undergoing orthodontic treatment was 68.4%. Conclusion: The incidence and prevalence rates of WSLs in patients undergoing orthodontic treatment are quite high and significant. This widespread problem of WSL development is an alarming challenge and warrants significant attention from both patients and providers, which should result in greatly increased emphasis on effective caries prevention. PMID:26759794

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

    SciTech Connect

    Liu Wangjing; Chang Yunshiang; Wang Chunghsiung; Kou, Guang-Hsiung; Lo Chufang . E-mail: gracelow@ntu.edu.tw

    2005-04-10

    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.

  10. [Antiparasitic effects of peracetic acid (PAA) against infective stages (theronts) of white spot disease, Ichthyophthirius multifiliis in vitro].

    PubMed

    Meinelt, T; Staaks, J; Staaks, G; Stüber, A; Bräunig, I

    2007-10-01

    White spot disease, caused by the protozoan parasite Ichthyophthirius multifiliis (I. multifiliis), invades nearly all fresh water fish species and causes huge economic losses. In Germany no protocide substance is legal for the treatment of I. multifilis. As an alternative substance the peracetic acid (PAA) was tested to treat the free invasive stage (theront) of the parasite. PAA concentrations of 0.3 ppm were able to kill all theronts in 120 min in our investigations. As a result of these investigations we recommend an interval-application of 0.3 to 0.5 ppm PAA for 30 to 150 min. This application should be prolonged for two life cycles of the parasite. Biotic parameters as e. g. fish species, and age as well as abiotic parameters as e. g. temperature, pH and organic load of the water could possibly influence the efficiency of the PAA application and should therefore be taken into account while picking the dosage and length of the PAA exposure.

  11. Identification of differentially expressed genes in shrimp (Penaeus stylirostris) infected with White spot syndrome virus by cDNA microarrays.

    PubMed

    Dhar, A K; Dettori, A; Roux, M M; Klimpel, K R; Read, B

    2003-12-01

    White spot syndrome virus (WSSV) is currently the most important viral pathogen infecting penaeid shrimp worldwide. Although considerable progress has been made in characterizing the WSSV genome and developing detection methods, information pertaining to host genes involved in WSSV pathogenesis is limited. We examined the potential of cDNA microarray analysis to study gene expression in WSSV-infected shrimp. Shrimp cDNAs were printed as low-density arrays on glass slides and were hybridized with Cy3/Cy5 labeled probes derived from RNA isolated from healthy and WSSV-infected shrimp. Genes that code for proteins that are relevant to crustacean immunity, structural proteins, as well as proteins of unknown function were among those whose mRNA expression was altered upon WSSV infection. To validate the microarray data, the temporal expression of three differentially expressed genes, an immune gene (C-type lectin-1), a structural gene (40S ribosomal protein), and a gene involved in lipid metabolism (fatty acid binding protein) was measured in healthy and WSSV-infected shrimp by real-time RT-PCR. The data suggest that WSSV infection alters the expression of a wide array of cellular genes, and provides a framework for further studies aimed at identifying genes whose function may provide insight into the mechanism of WSSV infection in shrimp.

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

    PubMed

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

    2011-03-01

    Outbreaks of white spot syndrome virus (WSSV) in shrimp culture and the relationship between the virus and virulence are not well understood. Here, we provide evidence showing that WSSV mixed-genotype infections correlate with lower outbreak incidence and that disease outbreaks correlate with single-genotype infections. We tested 573 shrimp samples from 81 shrimp ponds in the Mekong delta with outbreak or non-outbreak status. The variable number tandem repeat (VNTR) loci of WSSV were used as molecular markers for the characterization of single- and mixed-genotype infections. The overall prevalence of mixed-genotype WSSV infections was 25.7 %. Non-outbreak ponds had a significantly higher frequency of mixed-genotype infections than outbreak ponds for all VNTR loci, both at the individual shrimp as well as at the pond level. The genetic composition of WSSV populations appears to correlate with the health status of shrimp culture in ponds. The causal relationship between genotypic diversity and disease outbreaks can now be experimentally approached.

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

    PubMed

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

    2014-09-01

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

  14. Emilia sonchifolia extract activity against white spot syndrome virus and yellow head virus in shrimp cell cultures.

    PubMed

    Maikaeo, Lamai; Chotigeat, Wilaiwan; Mahabusarakam, Wilawan

    2015-07-23

    Emilia sonchifolia (L.) DC is a plant used in traditional medicine to treat several viral and bacterial diseases. The antiviral activities of selected Sephadex LH-20 column fractions and HPLC subfractions of an acetone extract of E. sonchifolia leaves were determined in shrimp Penaeus merguiensis primary lymphoid cells infected with either white spot syndrome virus (WSSV) or yellow head virus (YHV). WSSV and YHV replication was quantified using quantitative real-time PCR tests targeted to the VP19 and ORF1b gene transcripts, respectively. In lymphoid organ cells exposed to 100 µg ml⁻¹ of either the Sephadex fraction F14 or the HPLC F14 subfraction SF4, both fractions caused reduced replication, but YHV replication was reduced only by SF4. In the asthiazolyl blue mitochondrial enzyme activity assays to assess extract cytotoxicity, >60% of primary lymphoid organ cells remained viable following exposure to 100 µg ml⁻¹ of either F14 or SF4. GC-MS analysis of the HPLC F14 subfraction SF4 showed that it contained 2,4-di-tert-butylphenol. This study is the first to show that E. sonchifolia leaf extracts might be useful as bioactive agents to protect shrimp against viruses such as WSSV and YHV.

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

    PubMed

    Lu, Huasong; Ruan, Lingwei; Xu, Xun

    2011-10-25

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

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

    PubMed

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

    2010-11-15

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

  17. Dorsal transcription factor is involved in regulating expression of crustin genes during white spot syndrome virus infection.

    PubMed

    Huang, Xin; Wang, Wen; Ren, Qian

    2016-10-01

    Nuclear factor-kappa B (NF-κB) pathways play important roles in innate immune responses. In this study, we identified a dorsal homolog (MrDorsal) from freshwater prawn Macrobrachium rosenbergii. The full-length cDNA of MrDorsal comprised 2533 bp with an open reading frame of 1986 bp, which encoded a peptide of 661 amino acid residues. Amino acid sequence analysis showed that MrDorsal contains a Rel homolog domain and an IPT/TIG (i.e., Ig-like, plexin, and transcription factors) domain. The signature sequence of dorsal protein FRYMCEG existed in the deduced amino acid sequence. Sequence analysis showed that MrDorsal shared high similarities with Dorsal from invertebrate species. MrDorsal was abundant in the hemocytes and gills of healthy prawns but minute levels were detected in other tissues. The expression of MrDorsal was significantly upregulated 48 h after the white spot syndrome virus (WSSV-) challenge. Knockdown of MrDorsal using double-stranded RNA could suppress the transcription of crustin genes (MrCrustin2 and MrCrustin4) in gills of prawns after 48 h of the WSSV challenge. Results indicated that MrDorsal was involved to regulate the expression of crustin genes and it might play potential important roles during WSSV infection.

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

    PubMed

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

    2013-01-01

    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 (rVP28ΔN), C-terminal (rVP28ΔC) and middle (rVP28ΔM). 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.

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

    PubMed

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

    2015-10-01

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

  20. Molecular characterization, immune response against white spot syndrome virus infection of peroxiredoxin 4 in Fenneropenaeus chinensis and its antioxidant activity.

    PubMed

    Zhang, Qingli; Huang, Jie; Li, Fuhua; Liu, Shuang; Liu, Qinghui; Wei, Jiankai; Liang, Gaofeng; Xiang, Jianhai

    2014-03-01

    Peroxiredoxins (Prx) are a family of antioxidant proteins and perform important functions in intracellular signal transduction. Here, we report a Prx gene from Chinese shrimp Fenneropenaeus chinensis. The full-length cDNA of FcPrx gene contained an open reading frame of 735 bp encoding a polypeptide of 275 amino acids. The molecular mass of the deduced amino acid of FcPrx is 27445.43 Da with an estimated pI of 5.71. Sequence comparison showed that the FcPrx shares high identities with Prx IVs and it was named FcPrx4. A real-Time PCR (qRT-PCR) assay was developed to assess the mRNA expression of FcPrx4 in different tissues and temporal expression in hemocytes and hepatopancreas of F. chinensis challenged by white spot syndrome virus (WSSV). Transcripts of FcPrx4 can be detected in all tissues examined. The expression of FcPrx4 showed significant up-regulation in shrimp hemocytes and hepatopancreas after artificial infection with WSSV. A fusion protein containing FcPrx4 was produced in vitro and was confirmed by Matrix-assisted laser desorption ionization mass spectrometry (MALDI-MS) assay. And activity analysis indicated that the recombinant FcPrx4 proteins can reduce H2O2 in the presence of dithiothreitol.

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

    PubMed

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

    2015-10-01

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

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

    PubMed

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

    2014-01-01

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

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

    PubMed

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

    2012-12-01

    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.

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

    PubMed

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

    2015-10-20

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

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

    PubMed

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

    2014-11-01

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

  6. Comparative microarray profile of the hepatopancreas in the response of "Huanghai No. 2" Fenneropenaeus chinensis to white spot syndrome virus.

    PubMed

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

    2016-11-01

    White spot syndrome virus (WSSV) infects all shrimp species and is the greatest detriment to shrimp culture. To better understand the mechanism of molecular responses to WSSV infection in "Huanghai No. 2" Fenneropenaeus chinensis, a microarray technique was used. Microarray gene expression profiling of 59,137 unigenes identified Differentially Expressed Genes (DEGs) both in live and moribund shrimp at early, peak and late phases. In live shrimp, 1307, 1479 and 1539 DEGs were obtained in the early, peak and late phase, respectively. Meanwhile, 1536, 2181 and 1591 DEGs were obtained in moribund shrimp. Twenty known annotation genes are uniquely expressed in the late phase of live shrimp, including adhesion regulating molecule 1, arginine kinase, BUD31 homolog, and QM. Compared to WSSV-susceptible shrimp, 75 known annotation genes are uniquely expressed in WSSV-resistant shrimp, including arginine kinase, BUD31 homolog, clottable protein 2, caspase 2, cathepsin C, calnexin, HMGBb, Histone 3, and selenoprotein M. The gene expression patterns of the infected shrimp were altered by WSSV infection. To further confirm the expression of differentially expressed genes, real-time RT-PCR was performed to test six randomly selected genes. The data will provide valuable information to understand the immune mechanism of shrimp's response to WSSV.

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

    PubMed

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

    2015-08-01

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

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

    PubMed

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

    2015-01-01

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

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

    PubMed Central

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

    2017-01-01

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

  10. Label Free Detection of White Spot Syndrome Virus Using Lead Magnesium Niobate-Lead Titanate Piezoelectric Microcantilever Sensors

    PubMed Central

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

    2011-01-01

    We have investigated rapid, label free detection of white spot syndrome virus (WSSV) using the first longitudinal extension resonance peak of five lead-magnesium niobate-lead titanate (PMN-PT) piezoelectric microcantilever sensors (PEMS) 1050-700 μm long and 850-485 μm wide constructed from 8 μm thick PMN-PT freestanding films. The PMN-PT PEMS were encapsulated with a 3-mercaptopropltrimethoxysilane (MPS) insulation layer and further coated with anti-VP28 and anti-VP664 antibodies to target the WSSV virions and nucleocapsids, respectively. By inserting the antibody-coated PEMS in a flowing virion or nucleocapsid suspension, label-free detection of the virions and nucleocapsids were respectively achieved by monitoring the PEMS resonance frequency shift. We showed that positive label-free detection of both the virion and the nucleocapsid could be achieved at a concentration of 100 virions (nucleocapsids)/ml or 10 virions (nucleocapsids)/100μl, comparable to the detection sensitivity of polymerase chain reaction (PCR). However, in contrast to PCR, PEMS detection was label-free, in-situ and rapid (less than 30 min), potentially requiring minimal or no sample preparation. PMID:20863681

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

    PubMed

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

    2017-03-28

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

  12. A white spot syndrome virus microRNA promotes the virus infection by targeting the host STAT

    PubMed Central

    Ren, Qian; Huang, Ying; He, Yaodong; Wang, Wen; Zhang, Xiaobo

    2015-01-01

    JAK/STAT pathway plays an important role in invertebrates during virus infection. However the microRNA (miRNA)-mediated regulation of JAK/STAT is not intensively investigated. Viral miRNAs, encoded by virus genome, have emerged as important regulators in the virus-host interactions. In this study, a WSSV (white spot syndrome virus)-encoded miRNA (WSSV-miR-22) was characterized in shrimp during virus infection. The results showed that the viral miRNA could promote WSSV infection in shrimp by targeting the host STAT gene. When the expression of JAK or STAT was knocked down by sequence-specific siRNA, the WSSV copies in shrimp were significantly increased, indicating that the JAK/STAT played positive roles in the antiviral immunity of shrimp. The further findings revealed that TEP1 and TEP2 were the effectors of JAK-STAT signaling pathway. The silencing of TEP1 or TEP2 led to an increase of WSSV copies in shrimp, showing TEP1 and TEP2 were involved in the shrimp immune response against virus infection. Therefore our study presented a novel viral miRNA-mediated JAK/STAT-TEP1/TEP2 signaling pathway in virus infection. PMID:26671453

  13. Effects of competitor density and physical habitat structure on the competitive intensity of territorial white spotted charr Salvelinus leucomaenis.

    PubMed

    Hasegawa, K; Yamamoto, S

    2009-01-01

    This study compared the effects of interference competition in habitats of different complexity and in different densities. The influence of fish density and habitat structure was examined in manipulative experiments using young-of-the-year white spotted charr Salvelinus leucomaenis as a model species. The difference of specific growth rate (G) range, an index of interference competitive intensity, was significantly smaller in the structurally complex treatments than structurally simple treatments, while there were no significant difference between high-density and low-density treatments. Thus, physical habitat structure was more effective for mitigating interference competition than manipulating competitor density. Although interference competition was not affected by competitor density, mean G were suppressed in the high-density treatments. This implied that exploitative competition may cause the decrease of G rather than interference competition does in the high-density treatments. Mean G were also suppressed in the structurally complex treatments. Chaotic flow pattern created by physical habitat structures may decrease G by reducing foraging success of experimental fish in the complex treatments.

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

    PubMed Central

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

    2012-01-01

    A yearlong (September 2009–August 2010) study was undertaken to find out possible reasons for occasional occurrence of White Spot Syndrome Virus (WSSV) outbreak in the traditional prawn farms adjoining Cochin backwaters. Physicochemical and bacteriological parameters of water and sediment from feeder canal and four shrimp farms were monitored on a fortnightly basis. The physicochemical parameters showed variation during the two production cycles and between the farms studied. Dissolved 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. PMID:22593673

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

    PubMed

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

    2012-01-01

    A yearlong (September 2009-August 2010) study was undertaken to find out possible reasons for occasional occurrence of White Spot Syndrome Virus (WSSV) outbreak in the traditional prawn farms adjoining Cochin backwaters. Physicochemical and bacteriological parameters of water and sediment from feeder canal and four shrimp farms were monitored on a fortnightly basis. The physicochemical parameters showed variation during the two production cycles and between the farms studied. Dissolved 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.

  16. Growth and metabolism in the embryonic white-spotted bamboo shark, Chiloscyllium plagiosum: comparison with embryonic birds and reptiles.

    PubMed

    Tullis, A; Peterson, G

    2000-01-01

    Birds and reptiles have been important models for studying the energetics of embryonic development. Studies on these groups reveal three metabolic patterns: an exponential increase in metabolism with embryo age, a sigmoidal increase with age, or a sigmoidal increase followed by a decrease before hatching. Models developed to explain avian metabolic patterns and developmental costs partition total costs between growth and maintenance. To test the generality of these models, we examined embryonic energetics of the oviparous white-spotted bamboo shark Chiloscyllium plagiosum. Oviparous sharks must actively ventilate during development, which could increase their development costs relative to birds and reptiles. Our results demonstrated that bamboo shark embryos have a peaked metabolic pattern and sigmoidal increase in body mass similar to ratites, crocodilians, and some turtles. The total cost of development was higher in bamboo sharks than in reptiles and many birds. However, calculations reveal that the high cost of bamboo shark development can be explained by the relatively long incubation time rather than the additional cost of muscular movement. Finally, an avian model can reasonably describe shark embryonic metabolism, suggesting that movement costs do not significantly alter the metabolic pattern during development.

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

    PubMed

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

    2014-07-01

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

  18. The interaction of two-spotted spider mites, Tetranychus urticae Koch, with Cry protein production and predation by Amblyseius andersoni (Chant) in Cry1Ac/Cry2Ab cotton and Cry1F maize.

    PubMed

    Guo, Yan-Yan; Tian, Jun-Ce; Shi, Wang-Peng; Dong, Xue-Hui; Romeis, Jörg; Naranjo, Steven E; Hellmich, Richard L; Shelton, Anthony M

    2016-02-01

    Crops producing insecticidal crystal (Cry) proteins from the bacterium, Bacillus thuringiensis (Bt), are an important tool for managing lepidopteran pests on cotton and maize. However, the effects of these Bt crops on non-target organisms, especially natural enemies that provide biological control services, are required to be addressed in an environmental risk assessment. Amblyseius andersoni (Acari: Phytoseiidae) is a cosmopolitan predator of the two-spotted spider mite, Tetranychus urticae (Acari: Tetranychidae), a significant pest of cotton and maize. Tri-trophic studies were conducted to assess the potential effects of Cry1Ac/Cry2Ab cotton and Cry1F maize on life history parameters (survival rate, development time, fecundity and egg hatching rate) of A. andersoni. We confirmed that these Bt crops have no effects on the biology of T. urticae and, in turn, that there were no differences in any of the life history parameters of A. andersoni when it fed on T. urticae feeding on Cry1Ac/Cry2Ab or non-Bt cotton and Cry1F or non-Bt maize. Use of a susceptible insect assay demonstrated that T. urticae contained biologically active Cry proteins. Cry proteins concentrations declined greatly as they moved from plants to herbivores to predators and protein concentration did not appear to be related to mite density. Free-choice experiments revealed that A. andersoni had no preference for Cry1Ac/Cry2Ab cotton or Cry1F maize-reared T. urticae compared with those reared on non-Bt cotton or maize. Collectively these results provide strong evidence that these crops can complement other integrated pest management tactics including biological control.

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

    NASA Astrophysics Data System (ADS)

    Ohji, Madoka; Harino, Hiroya; Arai, Takaomi

    2011-01-01

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

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

    PubMed

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

    2016-01-01

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

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

    PubMed Central

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

    2016-01-01

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

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

    PubMed Central

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

    2014-01-01

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

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

    PubMed Central

    Fallahinejad Ghajari, Masoud; Eslamian, Ladan; Naji Rad, Azam; Morovati, Seyyedeh Pouya

    2015-01-01

    Objectives: This study aimed to compare the incidence of white spot lesions (WSLs) around orthodontic bands following the application of two glass ionomer (GI) cements namely GC Gold Label and GC Fuji Plus for six to 12 months. Materials and Methods: A total of 186 permanent first molars of orthodontic patients requiring banding of at least two permanent first molars were chosen. The teeth were examined for caries and presence of WSLs by visual inspection and by DIAGNOdent (scores 0–29). Orthodontic bands were randomly cemented to the right or left molars using GC Gold Label or GC Fuji Plus GI cements. Samples were randomly divided into three groups and bands were removed after six, nine and 12 months in groups 1, 2 and 3, respectively. The teeth were then examined for caries and presence of WSLs by visual inspection. DIAGNOdent was used on the buccal and lingual surfaces to determine the presence of WSLs. The data were statistically analyzed using one-way ANOVA, multivariate repeated measures ANOVA, the Kruskal Wallis and the Mann-Whitney tests. Results: Totally 174 teeth were evaluated. DIAGNOdent scores were not significantly different before cementation and after removal of bands in buccal and lingual surfaces of the teeth in the two cement groups. Lesions simulating WSLs were seen in 21 out of 174 teeth but DIAGNOdent scores were not indicative of caries. Conclusion: Remarkable WSLs were not detected visually or by DIAGNOdent at six, nine or 12 months following the cementation of bands with two GI cements. PMID:27559351

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

    PubMed Central

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

    2013-01-01

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

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

    PubMed

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

    2016-02-01

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

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

    PubMed

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

    2012-12-01

    Viral latency has been recently observed to be associated with White spot syndrome virus (WSSV) infection in shrimp. In the present study, shrimp samples (Penaeus monodon) surviving WSSV infection were examined for presence of WSSV in latent phase. Virus latency was observed in shrimp which were either experimentally challenged with WSSV and survived the infection or those which survived the natural infection. Three viral transcripts (ORFs 427, 151, 366) associated with latency were analyzed by real-time PCR. The shrimp surviving the natural WSSV infection on estimation with RT-PCR were found to have low grade of WSSV infection (less than 56 copies of WSSV). All the shrimp samples were RT-PCR negative for structural protein genes of WSSV, VP24 and VP28, indicating that these samples were harboring latent phase virus. RT-PCR of all the shrimp samples which survived WSSV infection revealed amplification of phagocytosis activating protein (PAP) gene (435 bp) with higher gene expression levels in experimentally challenged shrimp when compared to naturally infected shrimp. The expression of PAP in WSSV infected shrimp samples indicates its possible role in host response for resistance against WSSV infection. PAP was cloned and expressed as recombinant protein for protection studies. Shrimp were injected with three doses (5, 15 and 20 μg g(-1) body weight) of recombinant PAP. Relative percent survival of 10 % was observed in shrimp immunized with the dose of 15 μg g(-1) body weight of recombinant PAP. The expression of both WSSV latency associated and PAP genes obtained from shrimp surviving the WSSV infection, indicates the possible role of these genes in host-pathogen interaction.

  7. Enamel loss associated with orthodontic adhesive removal on teeth with white spot lesions: an in vitro study.

    PubMed

    Tüfekçi, Eser; Merrill, Thomas E; Pintado, Maria R; Beyer, John P; Brantley, William A

    2004-06-01

    Teeth with white spot lesions (WSL) might be more prone to enamel loss during bracket debonding. This in vitro study compared enamel loss from teeth with (n = 14) and without (n = 14) WSL after polishing with low-speed finishing burs or disks (Sof-Lex, 3M ESPE, St Paul, Minn). Debonded surfaces were analyzed with a contact stylus profilometer, and digitized data were compared with baseline readings by using AnSur NT software (Regents, University of Minnesota, Minneapolis, Minn). Specimen surfaces were also examined with a scanning electron microscope. Two-way analysis of variance was performed to analyze the data. In teeth without WSL, the volume losses were 0.16 mm(3) for the bur group and 0.10 mm(3) for the disk group; the mean maximum depths were 47.7 microm for the bur group and 54.3 microm for the disk group. In teeth with WSL, the volume losses were 0.06 and 0.17 mm(3), and the mean maximum depths were 35.1 and 48.7 microm for the bur and disk groups, respectively. There were no significant differences in enamel loss between the 2 groups of teeth without WSL (P =.12). However, in teeth with WSL, the burs removed less enamel than the disks (P = 0.006). Scanning electron microscope examination showed that any damage on the enamel surface was usually located in the cervical third of the teeth. On most specimens, even though tooth surfaces appeared resin-free to the naked eye, there were remnants of it. The differences between groups were so small that they might be clinically insignificant.

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

    PubMed

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

    2013-12-01

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

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

    PubMed

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

    2015-09-01

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

  10. Isolation and expression analysis of an MAPKK gene from Fenneropenaeus chinensis in response to white spot syndrome virus infection.

    PubMed

    Li, Xupeng; Kong, Jie; Meng, Xianhong; Luo, Kun; Luan, Sheng; Cao, Baoxiang; Liu, Ning

    2016-08-01

    Mitogen-activated kinase kinase (MAPKK) is an important gene involved in the host-virus interaction process. To obtain a better understanding of MAPKK in the interaction process between the Chinese shrimp Fenneropenaeus chinensis and white spot syndrome virus (WSSV), we cloned the sequence of an MAPKK cDNA from F. chinensis (FcMAPKK) and investigated the effect of FcMAPKK on WSSV infection. The results showed that the FcMAPKK gene contained a 1227 bp open reading frame (ORF), which encoded a highly conserved protein with a serine/threonine protein kinase catalytic (S_TKc) domain. The deduced amino acid sequence of FcMAPKK shared identities between 11.9 and 92.6% with MAPKKs from vertebrate, invertebrate, plant and fungus species. The FcMAPKK was expressed in all the examined tissues in the normal F. chinensis. FcMAPKK expression level was highest in the hepatopancreas where it was approximately 2.6-fold the expression level in the gill, and lowest in the muscle where it was approximately 0.3-fold the expression level in the hepatopancreas. The FcMAPKK expression levels in the muscle, gill, and hepatopancreas were all changed post WSSV challenge. The FcMAPKK expression was significantly (P < 0.01) up-regulated in the muscle of F. chinensis at 48 h post WSSV infection. The WSSV began to replicate quickly in the normal F. chinensis at 48 h post infection, while the WSSV replication in the U0126-treated F. chinensis could be significantly (P < 0.05) inhibited. The results suggested that FcMAPKK might be involved in the WSSV infection process, and hijacking of FcMAPKK might be required for WSSV replication in F. chinensis.

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

    PubMed

    Dieu, Bui Thi Minh; Marks, Hendrik; Zwart, Mark P; Vlak, Just M

    2010-05-01

    Variable genomic loci have been employed in a number of molecular epidemiology studies of white spot syndrome virus (WSSV), but it is unknown which loci are suitable molecular markers for determining WSSV spread on different spatiotemporal scales. Although previous work suggests that multiple introductions of WSSV occurred in central Vietnam, it is largely uncertain how WSSV was introduced and subsequently spread. Here, we evaluate five variable WSSV DNA loci as markers of virus spread on an intermediate (i.e. regional) scale, and develop a detailed and statistically supported model for the spread of WSSV. The genotypes of 17 WSSV isolates from along the coast of Vietnam--nine of which were newly characterized in this study--were analysed to obtain sufficient samples on an intermediate scale and to allow statistical analysis. Only the ORF23/24 variable region is an appropriate marker on this scale, as geographically proximate isolates show similar deletion sizes. The ORF14/15 variable region and variable-number tandem repeat (VNTR) loci are not useful as markers on this scale. ORF14/15 may be suitable for studying larger spatiotemporal scales, whereas VNTR loci are probably suitable for smaller scales. For ORF23/24, there is a clear pattern in the spatial distribution of WSSV: the smallest genomic deletions are found in central Vietnam, and larger deletions are found in the south and the north. WSSV genomic deletions tend to increase over time with virus spread in cultured shrimp, and our data are therefore congruent with the hypothesis that WSSV was introduced in central Vietnam and then radiated out.

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

    PubMed

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

    2015-04-01

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

  13. Interaction of the Small GTPase Cdc42 with Arginine Kinase Restricts White Spot Syndrome Virus in Shrimp.

    PubMed

    Xu, Ji-Dong; Jiang, Hai-Shan; Wei, Tian-Di; Zhang, Ke-Yi; Wang, Xian-Wei; Zhao, Xiao-Fan; Wang, Jin-Xing

    2017-03-01

    Many types of small GTPases are widely expressed in eukaryotes and have different functions. As a crucial member of the Rho GTPase family, Cdc42 serves a number of functions, such as regulating cell growth, migration, and cell movement. Several RNA viruses employ Cdc42-hijacking tactics in their target cell entry processes. However, the function of Cdc42 in shrimp antiviral immunity is not clear. In this study, we identified a Cdc42 protein in the kuruma shrimp (Marsupenaeus japonicus) and named it MjCdc42. MjCdc42 was upregulated in shrimp challenged by white spot syndrome virus (WSSV). The knockdown of MjCdc42 and injection of Cdc42 inhibitors increased the proliferation of WSSV. Further experiments determined that MjCdc42 interacted with an arginine kinase (MjAK). By analyzing the binding activity and enzyme activity of MjAK and its mutant, ΔMjAK, we found that MjAK could enhance the replication of WSSV in shrimp. MjAK interacted with the envelope protein VP26 of WSSV. An inhibitor of AK activity, quercetin, could impair the function of MjAK in WSSV replication. Further study demonstrated that the binding of MjCdc42 and MjAK depends on Cys(271) of MjAK and suppresses the WSSV replication-promoting effect of MjAK. By interacting with the active site of MjAK and suppressing its enzyme activity, MjCdc42 inhibits WSSV replication in shrimp. Our results demonstrate a new function of Cdc42 in the cellular defense against viral infection in addition to the regulation of actin and phagocytosis, which has been reported in previous studies. IMPORTANCE The interaction of Cdc42 with arginine kinase plays a crucial role in the host defense against WSSV infection. This study identifies a new mechanism of Cdc42 in innate immunity and enriches the knowledge of the antiviral innate immunity of invertebrates.

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

    PubMed

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

    2016-08-01

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

  15. Pupil - white spots

    MedlinePlus

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

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

    PubMed

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

    2014-12-01

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

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

    PubMed

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

    2015-01-01

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

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

    NASA Astrophysics Data System (ADS)

    Zhang, Xiaojing; Song, Xiaoling; Huang, Jie

    2016-11-01

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

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

    PubMed Central

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

    2015-01-01

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

  20. Resistance to Gray Leaf Spot of Maize: Genetic Architecture and Mechanisms Elucidated through Nested Association Mapping and Near-Isogenic Line Analysis

    PubMed Central

    Benson, Jacqueline M.; Poland, Jesse A.; Benson, Brent M.; Stromberg, Erik L.; Nelson, Rebecca J.

    2015-01-01

    Gray leaf spot (GLS), caused by Cercospora zeae-maydis and Cercospora zeina, is one of the most important diseases of maize worldwide. The pathogen has a necrotrophic lifestyle and no major genes are known for GLS. Quantitative resistance, although poorly understood, is important for GLS management. We used genetic mapping to refine understanding of the genetic architecture of GLS resistance and to develop hypotheses regarding the mechanisms underlying quantitative disease resistance (QDR) loci. Nested association mapping (NAM) was used to identify 16 quantitative trait loci (QTL) for QDR to GLS, including seven novel QTL, each of which demonstrated allelic series with significant effects above and below the magnitude of the B73 reference allele. Alleles at three QTL, qGLS1.04, qGLS2.09, and qGLS4.05, conferred disease reductions of greater than 10%. Interactions between loci were detected for three pairs of loci, including an interaction between iqGLS4.05 and qGLS7.03. Near-isogenic lines (NILs) were developed to confirm and fine-map three of the 16 QTL, and to develop hypotheses regarding mechanisms of resistance. qGLS1.04 was fine-mapped from an interval of 27.0 Mb to two intervals of 6.5 Mb and 5.2 Mb, consistent with the hypothesis that multiple genes underlie highly significant QTL identified by NAM. qGLS2.09, which was also associated with maturity (days to anthesis) and with resistance to southern leaf blight, was narrowed to a 4-Mb interval. The distance between major leaf veins was strongly associated with resistance to GLS at qGLS4.05. NILs for qGLS1.04 were treated with the C. zeae-maydis toxin cercosporin to test the role of host-specific toxin in QDR. Cercosporin exposure increased expression of a putative flavin-monooxygenase (FMO) gene, a candidate detoxification-related gene underlying qGLS1.04. This integrated approach to confirming QTL and characterizing the potential underlying mechanisms advances the understanding of QDR and will facilitate the

  1. Resistance to gray leaf spot of maize: genetic architecture and mechanisms elucidated through nested association mapping and near-isogenic line analysis.

    PubMed

    Benson, Jacqueline M; Poland, Jesse A; Benson, Brent M; Stromberg, Erik L; Nelson, Rebecca J

    2015-03-01

    Gray leaf spot (GLS), caused by Cercospora zeae-maydis and Cercospora zeina, is one of the most important diseases of maize worldwide. The pathogen has a necrotrophic lifestyle and no major genes are known for GLS. Quantitative resistance, although poorly understood, is important for GLS management. We used genetic mapping to refine understanding of the genetic architecture of GLS resistance and to develop hypotheses regarding the mechanisms underlying quantitative disease resistance (QDR) loci. Nested association mapping (NAM) was used to identify 16 quantitative trait loci (QTL) for QDR to GLS, including seven novel QTL, each of which demonstrated allelic series with significant effects above and below the magnitude of the B73 reference allele. Alleles at three QTL, qGLS1.04, qGLS2.09, and qGLS4.05, conferred disease reductions of greater than 10%. Interactions between loci were detected for three pairs of loci, including an interaction between iqGLS4.05 and qGLS7.03. Near-isogenic lines (NILs) were developed to confirm and fine-map three of the 16 QTL, and to develop hypotheses regarding mechanisms of resistance. qGLS1.04 was fine-mapped from an interval of 27.0 Mb to two intervals of 6.5 Mb and 5.2 Mb, consistent with the hypothesis that multiple genes underlie highly significant QTL identified by NAM. qGLS2.09, which was also associated with maturity (days to anthesis) and with resistance to southern leaf blight, was narrowed to a 4-Mb interval. The distance between major leaf veins was strongly associated with resistance to GLS at qGLS4.05. NILs for qGLS1.04 were treated with the C. zeae-maydis toxin cercosporin to test the role of host-specific toxin in QDR. Cercosporin exposure increased expression of a putative flavin-monooxygenase (FMO) gene, a candidate detoxification-related gene underlying qGLS1.04. This integrated approach to confirming QTL and characterizing the potential underlying mechanisms advances the understanding of QDR and will facilitate the

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

    PubMed

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

    2013-09-01

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

  3. Cloning of profilin (FcPFN) from the shrimp Fenneropenaeus chinensis, a highly expressed protein in white spot syndrome virus (WSSV)-infected shrimp.

    PubMed

    Kong, H J; Hong, G-E; Cho, H K; Nam, B-H; Kim, Y-O; Kim, W-J; Lee, S-J; Kim, K-K

    2009-01-01

    We isolated and characterized the profilin (FcPFN) cDNA from hemocytes of Fenneropenaeus chinensis, a unique shrimp species from the Yellow Sea. The FcPFN cDNA consists of 830 bp and encodes a polypeptide of 125 amino acids, having a predicted isoelectric point of 5.06. The deduced amino acid sequence of FcPFN shows 36% and 90% amino acid sequence identity to the profilin genes of Pacific white shrimp Litopenaeus vannamei and black tiger shrimp Penaeus monodon, respectively. The FcPFN mRNA was highly expressed in hemocytes and hepatopancreas and moderately in muscle of normal shrimp. The higher expression of FcPFN mRNA is observed in shrimp infected with the white spot syndrome virus (WSSV), which is a major concern in all shrimp-growing regions of the world. These results suggest a potential role for FcPFN in viral host defense mechanisms.

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

    PubMed

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

    2016-01-01

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

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

    USGS Publications Warehouse

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

    2014-01-01

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

  6. Protection against white spot syndrome virus (WSSV) infection in kuruma shrimp orally vaccinated with WSSV rVP26 and rVP28.

    PubMed

    Satoh, Jun; Nishizawa, Toyohiko; Yoshimizu, Mamoru

    2008-11-20

    White spot syndrome virus (WSSV) is the causative agent of white spot disease (WSD), one of the most serious diseases affecting global shrimp farming. We compared WSSV infection induction in kuruma shrimp Marsupenaeus japonicus by oral, immersion, and intramuscular injection (IM) exposure methods and evaluated the oral vaccine prepared from the recombinant WSSV proteins rVP26 and rVP28. The 50% lethal doses (LD50) of WSSV by oral, immersion, and IM challenges were 10(-0.4), 109-4.4), and 10(-7.7) g shrimp(-1), respectively, indicating that WSSV infection efficiency by oral challenge was significantly less than the other 2 challenge routes. However, in shrimp farms it is believed that WSSV infection is easily and commonly established by the oral route as a result of cannibalization of WSSV-infected shrimp. Kuruma shrimp vaccinated orally with WSSV rVP26 or rVP28 were challenged with WSSV by oral, immersion, and IM routes to compare protection efficacy. The relative percent survival values were 100% for oral challenge, 70 to 71% for immersion, and 34 to 61% for IM. Thus, the protection against WSSV-infection that was induced in kuruma shrimp by oral vaccination with rVP26 or rVP28 seemed equivalent to that obtained through IM vaccination.

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

    PubMed

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

    2014-05-01

    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.

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

    PubMed

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

    2015-01-01

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

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

    PubMed

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

    2013-09-01

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

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

    PubMed

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

    2012-08-27

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

  11. Inactivation of Anisakis simplex L3 in the flesh of white spotted conger (Conger myriaster) by high hydrostatic pressure and its effect on quality.

    PubMed

    Lee, Ki-Hoon; Park, Shin Young; Ha, Sang-Do

    2016-06-01

    Koreans consume much seafood; the country is surrounded on the east, west and south by the sea. Koreans have eaten raw sashimi for a long time. However, a concern in the raw sea food industry is that the parasitic nematode Anisakis simplex L3 occurs naturally in marine fish. Thus, the fishery industry needs a non-thermal processing method. High hydrostatic pressure (HPP) has been demonstrated to be effective. White spotted conger flesh containing 20 live larvae was exposed to different pressures (150 and 200 MPa for 1 and 5 min; 250 and 300 MPa each for 1 min). The viability of A. simplex L3 was significantly (p < 0.05) reduced in the flesh of white spotted conger by the stepwise increase of high pressure and time. The conditions required to eliminate A. simplex L3 were as follows: 200 MPa for 5 min or 300 MPa for 1 min. The flesh of the white spotted conger treated at 300 MPa for 1 min was whiter and yellower than untreated controls or that treated at 200 MPa for 5 min. No significant changes (p > 0.05) in any of the Hunter colour ('L', 'a' and 'b') values were found after HPP at 200 MPa for 5 min. The fresh treated at 300 MPa for 1 min scored < 4.0 (the defect limit of quality) of flavour, texture and overall acceptability in untrained sensory evaluation using a seven-point hedonic scale. However, the flesh treated at 200 MPa for 5 min scored > 5.0 ('like') for all sensory parameters. This study suggested that HPP at 200 MPa for 5 min could potentially be used for the inactivation of A. simplex L3 in raw fishery food products without any concomitant changes in their colour or sensory qualities.

  12. Indirect selection for resistance to ear rot and leaf diseases in maize lines using biplots.

    PubMed

    Pereira, G S; Camargos, R B; Balestre, M; Von Pinho, R G; C Melo, W M

    2015-09-21

    Leaf disease and ear rot have caused reductions in maize yield in Brazil and other producer countries. Therefore, the aims of this study were to analyze the association between husked ear yield and the severity of maize white spot, gray leaf spot, helminthosporium, and ear rot caused by Fusarium verticillioides and Diplodia maydis using biplots in a mixed-model approach. The responses of 238 lines introduced to Brazil and four controls were evaluated using an incomplete block design with three replicates in two locations: Lavras and Uberlândia, Minas Gerais, Brazil. Two experiments were conducted in each location, one with F. verticillioides and the other with D. maydis. The mixed models elucidated the relationship between yield, leaf disease, and ear disease. Significant genotype x environment and genotype x pathogen interactions were observed. In conclusion, husked ear yield is more associated with ear rot than with the leaf diseases evaluated, justifying the indirect selection for resistance to kernel rot in maize-F. verticillioides and maize-D. maydis pathosystems by yield evaluation.

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

    PubMed

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

    2012-05-01

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

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

    PubMed

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

    2016-07-01

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

  15. Competition of infectious hypodermal and haematopoietic necrosis virus (IHHNV) with white spot syndrome virus (WSSV) for binding to shrimp cellular membrane.

    PubMed

    Yan, D C; Huang, J; Yang, B; Sun, H S; Wang, Y Y; Liu, X

    2016-10-01

    Infectious hypodermal and haematopoietic necrosis virus (IHHNV) and white spot syndrome virus (WSSV) are two widespread shrimp viruses. The interference of IHHNV on WSSV was the first reported case of viral interference that involved crustacean viruses and has been subsequently confirmed. However, the mechanisms underlying the induction of WSSV resistance through IHHNV infection are practically unknown. In this study, the interference mechanisms between IHHNV and WSSV were studied using a competitive ELISA. The binding of WSSV and IHHNV to cellular membrane of Litopenaeus vannamei was examined. The results suggested that there existed a mutual competition between IHHNV and WSSV for binding to receptors present on cellular membrane of L. vannamei and that the inhibitory effects of WSSV towards IHHNV were more distinct than those of IHHNV towards WSSV.

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

    PubMed

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

    2015-03-01

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

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

    PubMed

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

    2017-03-27

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

  18. Assessment of home-based processing methods to reduce the phytate content and phytate/zinc molar ratio of white maize (Zea mays).

    PubMed

    Hotz, C; Gibson, R S

    2001-02-01

    Various methods of processing maize suitable for household use in rural Malawi, Central Africa, were investigated for their ability to reduce its phytate content and phytate/zinc molar ratio. These methods included fermentation, germination, and soaking. Penta- and hexainositol phosphates were measured by HPLC, and zinc was measured by atomic absorption spectrophotometry. Natural lactic fermentation of maize flour slurries resulted in 88% phytate retention compared to unprocessed, unrefined maize flour porridges, whereas lower phytate retention was observed when a starter culture (61%) or germinated flour (71%) was also used. Fermentation of cooked maize flour porridges with germinated flour added resulted in 54-85% retention of phytate compared to controls. Soaking maize flour or pounded maize and decanting excess water resulted in 43 and 49% retention of phytate, respectively. The latter soaking procedures were simple and effective and were suitable for household use in rural Malawian communities.

  19. Influence of boat noises on escape behaviour of white-spotted eagle ray Aetobatus ocellatus at Moorea Island (French Polynesia).

    PubMed

    Berthe, Cecile; Lecchini, David

    2016-02-01

    The present study tested different sounds that could disturb eagle rays (Aetobatus ocellatus) during their foraging activities at Moorea, French Polynesia. Results showed that artificial white sound and single-frequency tones (40 Hz, 600 Hz or 1 kHz) did not have an effect on rays (at least 90% of rays continued to forage over sand), while playbacks of boat motor sound significantly disturbed rays during foraging activity (60% exhibited an escape behaviour). Overall, our study highlighted the negative effect of boat noises on the foraging activity of eagle rays. These noises produced by boat traffic could, however, have some positive effects for marine aquaculture if they could be used as a deterrent to repel the eagle rays, main predators of the pearl oysters.

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

    PubMed

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

    2016-02-29

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

  1. Kinetic analysis of internalization of white spot syndrome virus by haemocyte subpopulations of penaeid shrimp, Litopenaeus vannamei (Boone), and the outcome for virus and cell.

    PubMed

    Tuan, V V; De Gryse, G M A; Thuong, K V; Bossier, P; Nauwynck, H J

    2016-12-01

    Little is known about the innate antiviral defence of shrimp haemocytes. In this context, the haemocytes of penaeid shrimp Litopenaeus vannamei (Boone) were separated by iodixanol density gradient centrifugation into five subpopulations (sub): sub 1 (hyalinocytes), sub 2 and 3 (prohyalinocytes), sub 4 (semigranulocytes) and sub 5 (granulocytes) and exposed to beads, white spot syndrome virus (WSSV) and ultraviolet (UV)-killed WSSV. In a first experiment, the uptake of beads, white spot syndrome virus (WSSV) and UV-killed WSSV by these different haemocyte subpopulations was investigated using confocal microscopy. Only haemocytes of sub 1, 4 and 5 were internalizing beads, WSSV and UV-killed WSSV. Beads were engulfed by a much larger percentage of cells (91.2 in sub 1; 84.1 in sub 4 and 58.1 in sub 5) compared to WSSV (9.6 in sub 1; 10.5 in sub 4 and 7.9 in sub 5) and UV-killed WSSV (12.9 in sub 1; 13.3 in sub 4; and 11.8 in sub 5). In a second experiment, it was shown that upon internalization, WSS virions lost their envelope most probably by fusion with the cellular membrane of the endosome (starting between 30 and 60 min post-inoculation) and that afterwards the capsid started to become disintegrated (from 360 min post-inoculation). Expression of new viral proteins was not observed. Incubation of haemocyte subpopulations with WSSV but not with UV-killed WSSV and polystyrene beads resulted in a significant drop in haemocyte viability. To find the underlying mechanism, a third experiment was performed in which haemocyte subpopulations were exposed to a short WSSV DNA fragment (VP19) and CpG ODNs. These small DNA fragments induced cell death. In conclusion, WSSV is efficiently internalized by hyalinocytes, semigranulocytes and granulocytes, after which the virus loses its envelope; as soon as the capsids start to disintegrate, cell death is activated, which in part may be explained by the exposure of viral DNA to cellular-sensing molecules.

  2. Spatial Epidemiology and Risk Factor Analysis of White Spot Disease in the Shrimp Farming Industry of Sinaloa, Mexico, from 2005 to 2011.

    PubMed

    Muniesa, A; Mardones, F O; Chávez, M C; Montoya, L; Cabanillas, J A; de Blas, I; Martínez-López, B

    2016-07-08

    White spot disease (WSD), caused by the white spot syndrome virus, is currently one of the primary causes of mortality and economic losses in the shrimp farming industry worldwide. In Mexico, shrimp production is one of the most important primary activities generating an annual income of USD 711 million. However, WSD introduction in 1999 had a devastating impact for the Mexican shrimp industry. The aim of this study was to characterize the WSD spatio-temporal patterns and to identify the primary risk factors contributing to WSD occurrence from 2005 to 2011 in Sinaloa, Mexico. We used data collected by the 'Comité Estatal de Sanidad Acuícola de Sinaloa' from 2005 to 2011 regarding WSD outbreaks as well as environmental, production and husbandry factors at farm level. The spatio-temporal patterns of WSD were described using space-time scan statistics. The effect of 52 variables on the time to WSD outbreak occurrence was assessed using a multivariable Cox proportional hazards model. Results reveal that WSD risk and survival time were not homogeneously distributed as suggested by the significant clusters obtained using the space-time permutation model and the space-time exponential model, respectively. The Cox model revealed that the first production cycle [hazard ratio (HR) = 11.31], changes from 1 to 1.4°C of temperature oscillation caused by 'El Niño'/'La Niña' events (HR = 1.44) and high average daily growths (HR = 1.26) were significantly associated with lower survival (i.e. shorter time to WSD outbreak) on farm. Conversely, shrimp weight at the moment of the outbreak (HR = 0.159), changes from -0.9 to -0.5°C of temperature oscillation caused by 'El Niño'/'La Niña' events (HR = 0.540), high superficial water temperature during the pound stocking (HR = 0.823) and high (>100) number of days of culture (HR = 0.830) were factors associated with higher survival. Results are expected to inform the design of risk-based, intervention strategies to

  3. A new shrimp peritrophin-like gene from Exopalaemon carinicauda involved in white spot syndrome virus (WSSV) infection.

    PubMed

    Wang, Liyan; Li, Fuhua; Wang, Bing; Xiang, Jianhai

    2013-09-01

    Peritrophin was first separated from insect peritrophic membrane (PM), and it played an important role in stimulating the digestion of food and protecting insects from invasion by microorganisms. In this study, a full-length cDNA of a new peritrophin-like protein (EcPT) was cloned from the ridgetail white shrimp Exopalaemon carinicauda, which was an excellent experimental animal for shrimp. The full length cDNA comprised 1235 bp including an 873 bp open reading frame encoding 291 amino acids. The deduced amino acid sequence contained a segment of signal peptide and three conserved chitin binding type 2 domains (ChtBD2) characterized by having a 6-cysteine motif. Tissue expression analysis revealed that EcPT was mainly expressed in stomach and gills, which were also the two main target tissues of WSSV infection. The transcription levels of EcPT in both stomach and gills were found to have significantly changed upon WSSV infection by real-time PCR. Silencing EcPT by dsRNA interference led to higher survival rate of shrimp against WSSV challenge, which suggested that EcPT might be involved in WSSV infection.

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

    PubMed

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

    2013-09-01

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

  5. Enhanced survival of shrimp, Penaeus (Marsupenaeus) japonicus from white spot syndrome disease after oral administration of recombinant VP28 expressed in Brevibacillus brevis.

    PubMed

    Caipang, Christopher Marlowe A; Verjan, Noel; Ooi, Ei Lin; Kondo, Hidehiro; Hirono, Ikuo; Aoki, Takashi; Kiyono, Hiroshi; Yuki, Yoshikazu

    2008-09-01

    White spot syndrome virus (WSSV) disease is a major threat to shrimp culture worldwide. Here, we assessed the efficacy of the oral administration of purified recombinant VP28, an envelope protein of WSSV, expressed in a Gram-positive bacterium, Brevibacillus brevis, in providing protection in shrimp, Penaeus japonicus, upon challenge with WSSV. Juvenile shrimp (2-3g in body weight) fed with pellets containing purified recombinant VP28 (50microg/shrimp) for 2 weeks showed significantly higher survival rates than control groups when challenged with the virus at 3 days after the last day of feeding. However, when shrimp were challenged 2 weeks after the last day of feeding, survival rates decreased (33.4% and 24.93%, respectively). Survival rate was dose-dependent, increasing from 60.7 to 80.3% as the dose increased from 1 to 50microg/shrimp. At a dose of 50microg/shrimp, the recombinant protein provided protection as soon as 1 day after feeding (72.5% survival). Similar results were obtained with larger-sized shrimp. These results show that recombinant VP28 expressed in a Gram-positive bacterium is a potential oral vaccine against WSSV.

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

    PubMed

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

    2012-06-13

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

  7. Two new anti-apoptotic proteins of white spot syndrome virus that bind to an effector caspase (PmCasp) of the giant tiger shrimp Penaeus (Penaeus) monodon.

    PubMed

    Lertwimol, Tareerat; Sangsuriya, Pakkakul; Phiwsaiya, Kornsunee; Senapin, Saengchan; Phongdara, Amornrat; Boonchird, Chuenchit; Flegel, Timothy W

    2014-05-01

    White spot syndrome virus proteins WSSV134 and WSSV322 have been shown to bind with the p20 domain (residues 55-214) of Penaeus monodon caspase (PmCasp) protein through yeast two-hybrid screening. Binding was confirmed for the p20 domain and the full-length caspase by co-immunoprecipitation. WSSV134 is also known as the WSSV structural protein VP36A, but no function or conserved domains have been ascribed to WSSV322. Discovery of the caspase binding activity of these two proteins led to an investigation of their possible anti-apoptotic roles. Full-length PmCasp was confirmed to be an effector caspase by inducing apoptosis in transfected Sf-9 cells as assessed by DAPI staining. Using the same cell model, comparison of cells co-transfected with PmCasp and either WSSV134 or WSSV322 revealed that both of the binding proteins had anti-apoptotic activity. However, using the same Sf-9 protocol with anti-apoptosis protein-1 (AAP-1; also called WSSV449) previously shown to bind and inactivate a different effector caspase from P. monodon (Pm caspase) did not block apoptosis induced by PmCasp. The results revealed diversity in effector caspases and their viral protein inhibitors in P. monodon.

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

    PubMed

    Zhu, Fei; Zhang, Xiaobo

    2012-02-01

    White spot syndrome virus (WSSV) is a major shrimp viral pathogen responsible for large economic losses to shrimp aquaculture all over the world. The RNAi mediated by siRNA contributes a new strategy to control this viral disease. However, the efficient approach to deliver the siRNA into shrimp remains to be addressed. In this investigation, an antiviral vp28-siRNA was encapsulated in β-1,3-D-glucan, and then the β-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.

  9. PmVRP15, a novel viral responsive protein from the black tiger shrimp, Penaeus monodon, promoted white spot syndrome virus replication.

    PubMed

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

    2014-01-01

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

  10. Co-interactive DNA-binding between a novel, immunophilin-like shrimp protein and VP15 nucleocapsid protein of white spot syndrome virus.

    PubMed

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

    2011-01-01

    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 the WSSV genome during virion formation. However, little is known in its relationship with shrimp host cells. Using the yeast two-hybrid approach to screen a shrimp lymphoid organ (LO) cDNA library for proteins that might interact with VP15, a protein named PmFKBP46 was identified. It had high sequence similarity to a 46 kDa-immunophilin called FKBP46 from the lepidopteran Spodoptera frugiperda (the fall armyworm). The full length PmFKBP46 consisted of a 1,257-nucleotide open reading frame with a deduced amino acid sequence of 418 residues containing a putative FKBP-PPIase domain in the C-terminal region. Results from a GST pull-down assay and histological co-localization revealed that VP15 physically interacted with PmFKBP46 and that both proteins shared the same subcellular location in the nucleus. An electrophoretic mobility shift assay indicated that PmFKBP46 possessed DNA-binding activity and functionally co-interacted with VP15 in DNA binding. The overall results suggested that host PmFKBP46 might be involved in genome packaging by viral VP15 during virion assembly.

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

    PubMed

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

    2014-05-01

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

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

    PubMed

    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

    2013-03-01

    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 α-helix of PmRab7 (encompassing Ser74, Ile143, Thr184, Arg53, Asn144, Thr184, Arg53, Arg79) interacts with β-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.

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

    PubMed Central

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

    2016-01-01

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

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

    SciTech Connect

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

    2006-01-01

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

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

    SciTech Connect

    Wang Yuzhen; Zhang Xiaohua; Yuan Li; Xu Tao; Rao Yu; Li Jia; Dai Heping

    2008-08-08

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

  16. Analysis of the VP19 and VP28 genes of white spot syndrome virus in Korea and comparison with strains from other countries.

    PubMed

    Park, Jung-Eun; Shin, Hyun-Jin

    2009-01-01

    The results of our DNA analysis showed that there was 100% homology of the VP28 and VP19 genes of white spot syndrome virus (WSSV) among the samples collected from different shrimp farms in Korea. Comparing with an earlier isolated Korean strain, Korea01, the genes nucleotide sequences had only a single base difference which was observed in both VP19 and VP28. This resulted in a single amino acid substitution at position 40 of the latter. This implies that a single genetic strain of WSSV has been circulating in Korea and that its mutation rate is very low. In comparison with known sequences of VP19 and VP28 genes of WSSV isolates from other countries, the Korean strains had more than 99% sequence homology with those in gene and protein. Based on our sequence analysis of VP19 and VP28 of WSSV from various shrimp farms in Korea, the WSSV strains circulating in the region were genetically identical and similar to the strain identified two years ago. In addition, the Korean strain had close genetic identity with strains circulating in other Asian countries as well as other continents.

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

    PubMed

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

    2016-10-01

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

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

    PubMed

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

    2016-03-01

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

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

    PubMed

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

    2014-10-01

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

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

    PubMed

    Yingvilasprasert, Wanchart; Supungul, Premruethai; Tassanakajon, Anchalee

    2014-02-01

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

  1. Cloning and characterization of cytoplasmic dynein intermediate chain in Fenneropenaeus chinensis and its essential role in white spot syndrome virus infection.

    PubMed

    Feng, Jixing; Tang, Xiaoqian; Zhan, Wenbin

    2014-08-01

    To investigate the role of cytoplasmic dynein in white spot syndrome virus (WSSV) infection, the full-length cDNA of cytoplasmic dynein intermediate chain (FcDYNCI) was cloned in Fenneropenaeus chinensis, which consists of 2582 bp and encodes a polypeptide of 660 amino acids. Sequence analysis and multiple sequence alignment displayed that FcDYNCI was a member of cytoplasmic dynein 1 family. The FcDYNCI mRNA was most highly expressed in hemocytes, which was significantly up-regulated post WSSV infection. At 12 h post infection (hpi), confocal microscopic observation showed that WSSV could be co-localized with cytoplasmic dynein in hemocytes. After silencing by specific FcDYNCI dsRNA, the FcDYNCI mRNA level and the protein amount of FcDYNCI in hemocytes both exhibited a significant reduction, and the expression levels of three WSSV genes ie1, wsv477 and vp28 all exhibited the greatest decreases at 24 hpi. These results suggested that cytoplasmic dynein was involved in WSSV infection.

  2. The expression and purification of WSSV134 from white spot syndrome virus and its inhibitory effect on caspase activity from Penaeus monodon.

    PubMed

    Bowornsakulwong, Thanunthon; Charoensapsri, Walaiporn; Rattanarojpong, Triwit; Khunrae, Pongsak

    2017-02-01

    WSSV134 or VP36A protein of white spot syndrome virus was previously reported to be able to reduce apoptosis in Sf-9 cells transfected with caspase of Penaeus monodon (PmCasp). The protein was therefore believed to have a role in supporting the survival of WSSV inside the host cells during infection. However, the anti-apoptosis activity of WSSV134 involved in the inhibition of PmCasp is still unclear. In this study, we produced a recombinant WSSV134 (rWSSV134) and tested for its ability to inhibit PmCasp in vitro. The results from a caspase inhibition assay revealed that rWSSV134 could inhibit PmCasp in a dose-dependent manner. Since WSSV134 was predicted to contain three potential caspase binding sites, corresponding to the D54, D104 and D259, we then employed site-directed mutagenesis to investigate the involvement of these sites in PmCasp inhibition. D54A and D259A mutants could still inhibit PmCasp while D104A mutant lacks this activity. Our results confirmed that the WSSV134 is an inhibitor for PmCasp and that residue D104 is important for PmCasp inhibition.

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

    PubMed

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

    2015-03-01

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

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

    PubMed

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

    2015-05-01

    White spot syndrome virus (WSSV) is the most devastating pathogen of penaeid shrimp. While developing technology to vaccinate shrimp against WSSV, it is imperative to look into the immune response of the animal at molecular level. However, very little information has been generated in this direction. The present study is an attempt to understand the expression of bio-defense genes in gill tissues of Penaeus monodon in response to formalin inactivated WSSV. A WSSV vaccine with a viral titer of 1×10(9) DNA copies was prepared and orally administered to P. monodon at a rate of 1.75×10(6) DNA copies of inactivated virus preparation (IVP) day(-1) for 7days. The animals were challenged with WSSV on 1st and 5th day post vaccination, and temporal expression of bio-defense genes in gill tissues was studied. Survival of 100% and 50% were observed respectively on 1st and 5th day post vaccination challenge. The humoral immune genes prophenoloxidase (proPO), alpha 2-macroglobulin (α2M), crustin and PmRACK, and the cell mediated immune genes caspase and Rab7 were up regulated in gill tissue upon vaccination and challenge. The expression of humoral gene crustin and cellular gene Rab7 was related to survival in IVP administered shrimp. Results of the study suggest that these genes have roles in protecting shrimp from WSSV on vaccination.

  5. Shrimp STAT was hijacked by white spot syndrome virus immediate-early protein IE1 involved in modulation of viral genes.

    PubMed

    Yao, Defu; Ruan, Lingwei; Lu, Huasong; Shi, Hong; Xu, Xun

    2016-12-01

    STATs are a family of transcription factors that regulate a cascade of cellular processes including cell growth, differentiation, apoptosis and immune responses. However, they are usually targeted by viruses to assist infection. In this study, we identified that white spot syndrome virus (WSSV) immediate-early protein IE1 interacted with Litopenaeus vannamei STAT (LvSTAT) and thereby led to its phosphorylation activation. In addition, we demonstrated that LvSTAT could bind to the promoters of the viral immediate-early genes wsv051 and ie1 through STAT-binding motifs in vitro and vivo, allowing the enhancement of their promoters' activities. Moreover, IE1 could promote the transcriptional activation activity of LvSTAT to augment the transcription of wsv051 and ie1. In conclusion, our findings revealed a novel linkage between WSSV IE1 and shrimp STAT, which was a clue to well understand how WSSV adopted the active strategies to modulate the shrimp signaling pathway.

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

    PubMed

    Wongprasert, Kanokpan; Rudtanatip, Tawut; Praiboon, Jantana

    2014-01-01

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

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

    PubMed

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

    2013-12-01

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

  8. The cDNA Sequence of Two Hemocyanin Subunits from Red Swamp Crayfish Procambarus clarkii and their Responses to White Spot Syndrome Virus Infection.

    PubMed

    Zeng, Yong

    2016-03-01

    Hemocyanin, the respiratory protein of crustaceans, participates in the innate immune defense in these organisms. We cloned two hemocyanin subunit genes (PcHc1 and PcHc2), by using a degenerate primer PCR and rapid amplification of cDNA ends (RACE) approach, from the hepatopancreas of red swamp crayfish Procambarus clarkii. The transcripts of these two subunits were only detected in the hepatopancreas by reverse transcriptase quantitative PCR (RT-qPCR) analysis. The neighbor-joining and maximum parsimony phylogenetic analyses indicated that PcHc2 associated with a clade belong to the α-type hemocyanins and PcHc1 associated with another clade belonging to the β-type hemocyanins. The data obtained from the RT-qPCR indicated that the mRNA expression levels of these subunit genes followed almost the same regulation pattern in the crayfish challenged with white spot syndrome virus (WSSV). The fluctuation of mRNA expression levels of these two subunits after the WSSV challenge indicated that both of them may participate in the antiviral immune response of crayfish. Received April 12, 2015; accepted November 22, 2015.

  9. Real-time target-specific detection of loop-mediated isothermal amplification for white spot syndrome virus using fluorescence energy transfer-based probes.

    PubMed

    Chou, Pin-Hsing; Lin, Yu-Chan; Teng, Ping-Hua; Chen, Chu-Liang; Lee, Pei-Yu

    2011-04-01

    Aiming to establish a target amplicon-specific detection system for loop-mediated isothermal amplification (LAMP), the fluorescent resonance energy transfer (FRET) probe technology was applied to develop the FRET LAMP platform. This report describes the development of the first FRET LAMP assay targeting white spot syndrome virus (WSSV) of penaeid shrimp. A successful accelerated WSSV LAMP was assembled first in a conventional oven and confirmed by gel electrophoresis and dot-blot hybridization. Subsequently, two additional FRET probes designed to target one loop region within WSSV LAMP amplicons were added to the same LAMP reaction. The reactions were carried out in a LightCycler (Roche) and significant FRET signals were detected in real time. Optimization of the reaction using plasmid DNA shortened the time for the detection of 10(2) copies of the target DNA to less than 70min. Cross reactivity was absent with WSSV-free or infectious hypodermal and hematopoietic necrosis virus-infected Penaeus vannamei samples. The performance of this system was comparable with that of a nested PCR assay from 21 WSSV-infected shrimp. Specifically detecting target amplicons and requiring no post-amplification manipulation, the novel FRET LAMP assay should allow indisputable detection of pathogens with minimized risks of amplicon contamination.

  10. A 12-Week Assessment of the Treatment of White Spot Lesions with CPP-ACP Paste and/or Fluoride Varnish

    PubMed Central

    Güçlü, Zeynep Aslı; Alaçam, Alev

    2016-01-01

    This 12-week clinical study evaluated the impact of 10% CPP-ACP and 5% sodium fluoride varnish regimes on the regression of nonorthodontic white spot lesions (WSLs). The study included 21 children with 101 WSLs who were randomised into four treatment regimes: weekly clinical applications of fluoride varnish for the first month (FV); twice daily self-applications of CPP-ACP paste (CPP-ACP); weekly applications of fluoride varnish for the first month and twice daily self-applications of CPP-ACP paste (CPP-ACP-FV); and no intervention (control). All groups undertook a standard oral hygiene protocol and weekly consultation. Visual appraisals and laser fluorescence (LF) measurements were made in weeks one and twelve. The majority of WSLs in the control and FV groups exhibited no shift in appearance, whereas, in the CPP-ACP and CPP-ACP-FV groups, the lesions predominantly regressed. The visual and LF assessments indicated that the extent of remineralisation afforded by the treatments was of the following order: control ~ FV < CPP-ACP ~ CPP-ACP-FV. Self-applications of CPP-ACP paste as an adjunct to standard oral hygiene significantly improved the appearance and remineralisation of WSLs. No advantage was observed for the use of fluoride varnish as a supplement to either the standard or CPP-ACP-enhanced oral hygiene regimes. PMID:27843950

  11. Maize databases

    Technology Transfer Automated Retrieval System (TEKTRAN)

    This chapter is a succinct overview of maize data held in the species-specific database MaizeGDB (the Maize Genomics and Genetics Database), and selected multi-species data repositories, such as Gramene/Ensembl Plants, Phytozome, UniProt and the National Center for Biotechnology Information (NCBI), ...

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

    PubMed

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

    2015-01-01

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

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

    PubMed Central

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

    2001-01-01

    A rapid and highly sensitive real-time PCR detection and quantification method for infectious hypodermal and hematopoietic necrosis virus (IHHNV), a single-stranded DNA virus, and white spot virus (WSV), a double-stranded DNA (dsDNA) virus infecting penaeid shrimp (Penaeus sp.), was developed using the GeneAmp 5700 sequence detection system coupled with SYBR Green chemistry. The PCR mixture contains a fluorescence dye, SYBR Green, which upon binding to dsDNA exhibits fluorescence enhancement. The enhancement of fluorescence was proportional to the initial concentration of the template DNA. A linear relationship was observed between the amount of input plasmid DNA and cycle threshold (CT) values over a range of 1 to 105 copies of the viral genome. To control the variation in sampling and processing among samples, the shrimp β-actin gene was amplified in parallel with the viral DNA. The CT values of IHHNV- and WSV-infected samples were used to determine absolute viral copy numbers from the standard CT curves of these viruses. For each virus and its β-actin control, the specificity of amplification was monitored by using the dissociation curve of the amplified product. Using genomic DNA as a template, SYBR Green PCR was found to be 100- to 2000-fold more sensitive than conventional PCR, depending on the virus, for the samples tested. The results demonstrate that SYBR Green PCR can be used as a rapid and highly sensitive detection and quantification method for shrimp viruses and that it is amenable to high-throughout assay. PMID:11474000

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

    PubMed

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

    2013-01-01

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

  15. SUMO-Conjugating Enzyme E2 UBC9 Mediates Viral Immediate-Early Protein SUMOylation in Crayfish To Facilitate Reproduction of White Spot Syndrome Virus

    PubMed Central

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

    2013-01-01

    Successful viruses have evolved superior strategies to escape host defenses or exploit host biological pathways. Most of the viral immediate-early (ie) genes are essential for viral infection and depend solely on host proteins; however, the molecular mechanisms are poorly understood. In this study, we focused on the modification of viral IE proteins by the crayfish small ubiquitin-related modifier (SUMO) and investigated the role of SUMOylation during the viral life cycle. SUMO and SUMO ubiquitin-conjugating enzyme 9 (UBC9) involved in SUMOylation were identified in red swamp crayfish (Procambarus clarkii). Both SUMO and UBC9 were upregulated in crayfish challenged with white spot syndrome virus (WSSV). Replication of WSSV genes increased in crayfish injected with recombinant SUMO or UBC9, but injection of mutant SUMO or UBC9 protein had no effect. Subsequently, we analyzed the mechanism by which crayfish SUMOylation facilitates WSSV replication. Crayfish UBC9 bound to all three WSSV IE proteins tested, and one of these IE proteins (WSV051) was covalently modified by SUMO in vitro. The expression of viral ie genes was affected and that of late genes was significantly inhibited in UBC9-silenced or SUMO-silenced crayfish, and the inhibition effect was rescued by injection of recombinant SUMO or UBC9. The results of this study demonstrate that viral IE proteins can be modified by crayfish SUMOylation, prompt the expression of viral genes, and ultimately benefit WSSV replication. Understanding of the mechanisms by which viruses exploit host components will greatly improve our knowledge of the virus-host “arms race” and contribute to the development of novel methods against virulent viruses. PMID:23097446

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

    PubMed

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

    2016-04-01

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

  17. Blocking the large extracellular loop (LEL) domain of FcTetraspanin-3 could inhibit the infection of white spot syndrome virus (WSSV) in Chinese shrimp, Fenneropenaeus chinensis.

    PubMed

    Gui, Lang; Wang, Bing; Li, Fu-Hua; Sun, Yu-Miao; Luo, Zhan; Xiang, Jian-Hai

    2012-06-01

    Tetraspanins belong to the transmembrane 4 superfamily (TM(4)SF), which span the cell membrane 4 times and act as bridges or connectors. Increasing evidences have shown that tetraspanins play important role in virus infection. The large extracellular loop (LEL) of a tetraspanin is considered as a possible target of some virus. Tetraspanins are widely found in invertebrates, but the functional roles of most invertebrate tetraspanins have remained unknown. Recently, a tetraspanin, called FcTetraspanin-3, was cloned from the cDNA library of Chinese shrimp, Fenneropenaeus chinensis. The FcTetraspanin-3 constitutive expression in all examined tissues and the expression of the gene were highly induced in hepatopancreas, lymphoid organ and intestine by white spot syndrome virus (WSSV) challenge. In this study, we expressed and purified the recombinant peptide containing the LEL domain of FcTetraspanin-3, and produced the anti-LEL polyclone antibody. The expression of FcTetraspanin-3 was observed by real-time PCR and Western blot. Also, the localization of FcTetraspanin-3-positive cells in intestine and hepatopancreas were revealed by immunofluorescence. The results of anti-LEL antibody blocking experiments shown that the antibody can significantly reduce the mortality of shrimp challenged by WSSV. Additionally, dsRNA interference was utilized to examine the functional role of FcTetraspanin-3 in response to WSSV infection, and a sensible decrease of the viral copy number in the tetraspanin knockdown shrimp. These results suggested the blocking of LEL domain of FcTetraspanin-3 could inhibit the infection of WSSV. FcTetraspanin-3 might play an important role in response to WSSV infection, and the LEL domain of FcTetraspanin-3 might mediate the entry of WSSV.

  18. Expression Profile of Penaeus monodon Ubiquitin Conjugating Enzyme (PmUbc) at Protein Level in White spot syndrome virus Challenged Shrimp.

    PubMed

    Keezhedath, Jeena; Kurcheti, Pani Prasad; Pathan, Mujahid Khan; Babu, Gireesh P; Tripathi, Gayatri; Sudhagar, Arun; Rao, Srinivas P

    2013-06-01

    White spot syndrome virus (WSSV) is one of the major pathogens in shrimp aquaculture. Four proteins of WSSV are predicted to encode a RING H2 domain, which in presence of ubiquitin conjugating enzyme (E2) in shrimps can function as viral E3 ligase and modulate the host ubiquitin proteasome pathway. Modulation of host ubiquitin proteasome pathway by viral proteins is implicated in viral pathogenesis. In the present study, expression profile of Penaeus monodon Ubiquitin conjugating enzyme (PmUbc) was studied at protein level in WSSV challenged shrimp. A time point analysis of the expression of PmUbc was carried out at 0, 3, 6, 12, 24, 48 and 72 h post WSSV challenge in P. monodon. Recombinant PmUbc (rPmUbc) was produced in prokaryotic expression vector, BL21 (DE3) pLys S. The PmUbc expression pattern was studied by ELISA with rPmUbc antibodies raised in rabbit. A significant increase in PmUbc expression at 24 h post infection (hpi) was observed followed by a decline till 72 hpi. Since the up-regulation and a tremendous decline of PmUbc protein expression was observed at 24 and in 72 hpi respectively in ELISA, it can be speculated that these proteins might interact with host ubiquitination pathway for viral pathogenesis. Many findings have shown that viral infection can up-regulate expression of ubiquitin and that the ubiquitin system plays a key role in the course of viral infection. The present study reveals the expression patterns of PmUbc at protein level in WSSV infected P. monodon. However, further studies are to be carried out to unfold the molecular mechanism of interaction between host and virus to devise efficient control strategies for this major culprit in shrimp culture industry.

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

    PubMed Central

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

    2013-01-01

    In response to endoplasmic reticulum (ER) stress, the signaling pathway termed unfolded protein response (UPR) is activated. To investigate the role of UPR in Litopenaeus vannamei immunity, the activating transcription factor 4 (designated as LvATF4) which belonged to a branch of the UPR, the [protein kinase RNA (PKR)-like ER kinase, (PERK)]-[eukaryotic initiation factor 2 subunit alpha (eIF2α)] 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

  20. Marine derived compounds as binders of the White spot syndrome virus VP28 envelope protein: In silico insights from molecular dynamics and binding free energy calculations.

    PubMed

    Sivakumar, K C; Sajeevan, T P; Bright Singh, I S

    2016-10-01

    White spot syndrome virus (WSSV) remains as one of the most dreadful pathogen of the shrimp aquaculture industry owing to its high virulence. The cumulative mortality reaches up to 100% within in 2-10days in a shrimp farm. Currently, no chemotherapeutics are available to control WSSV. The viral envelope protein, VP28, located on the surface of the virus particle acts as a vital virulence factor in the initial phases of inherent WSSV infection in shrimp. Hence, inhibition of envelope protein VP28 could be a novel way to deal with infection by inhibiting its interaction in the endocytic pathway. In this direction, a timely attempt was made to recognize a potential drug candidate of marine origin against WSSV using VP28 as a target by employing in silico docking and molecular dynamic simulations. A virtual library of 388 marine bioactive compounds was extracted from reports published in Marine Drugs. The top ranking compounds from docking studies were chosen from the flexible docking based on the binding affinities (ΔGb). In addition, the MD simulation and binding free energy analysis were implemented to validate and capture intermolecular interactions. The results suggested that the two compounds obtained a negative binding free energy with -40.453kJ/mol and -31.031kJ/mol for compounds with IDs 30797199 and 144162 respectively. The RMSD curve indicated that 30797199 moves into the hydrophobic core, while the position of 144162 atoms changes abruptly during simulation and is mostly stabilized by water bridges. The shift in RMSD values of VP28 corresponding to ligand RMSD gives an insight into the ligand induced conformational changes in the protein. This study is first of its kind to elucidate the explicit binding of chemical inhibitor to WSSV major structural protein VP28.

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

    PubMed

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

    2015-04-01

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

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

    PubMed

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

    2017-03-01

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

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

    PubMed

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

    2016-12-01

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

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

    PubMed

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

    2015-05-01

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

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

    PubMed

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

    2011-09-09

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

  6. Anti-melanization mechanism of the white spot syndrome viral protein, WSSV453, via interaction with shrimp proPO-activating enzyme, PmproPPAE2.

    PubMed

    Sutthangkul, Jantiwan-; Amparyup, Piti-; Eum, Jai Hoon; Strand, Michael R; Tassanakajon, Anchalee

    2017-01-28

    Inhibition of the host melanization reaction, activated by the prophenoloxidase activating (proPO) system, is one of the crucial evasion strategies of pathogens. Recently, the shrimp pathogen, white spot syndrome virus (WSSV), was found to inhibit melanization in the shrimp, Penaeus monodon. The viral protein WSSV453 was previously shown to interact with PO-activating enzyme 2 (PmPPAE2) and reported to be involved in suppressing the shrimp melanization response after WSSV infection. Here, we characterized how WSSV453 inhibits melanization. WSSV453 is a non-structural viral protein, which was first detected in shrimp hemocytes at 6 hours post infection (hpi) by WSSV and in shrimp plasma at 24 hpi. We produced recombinant proteins for three components of the P. monodon, proPO system: PmproPPAE2, PmproPO1 and PmproPO2. Functional assays showed that active PmPPAE2 processed PmproPO1 and 2 to produce functional PO. Incubation of WSSV453 with PmproPPAE2 dose-dependently reduced PmPPAE2 activity toward PmPO1 or PmPO2. In contrast, WSSV453 had no effect on activated PmPPAE2. The addition of active PmPPAE2 to WSSV-infected shrimp plasma at day 2 post-infection also rescued PO activity. Taken together, these results indicate that the anti-melanization activity of WSSV is due to WSSV453, which interacts with PmproPPAE2 and interferes its activation to active PmPPAE2.

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

    PubMed

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

    2015-09-01

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

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

    SciTech Connect

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

    2007-01-01

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

  9. Liver spots

    MedlinePlus

    Sun-induced skin changes - liver spots; Senile or solar lentigines; Skin spots - aging; Age spots ... Liver spots are changes in skin color that occur in older skin. The coloring may be due to aging, exposure to the sun or other sources of ...

  10. White noise effects of U.S. crude oil spot prices on stock prices of a publicly traded company: A case study cross-correlation analysis based on green energy management theory

    NASA Astrophysics Data System (ADS)

    Roberts, Peter M.

    The purpose of this study was to examine white noise effects of U.S. crude oil spot prices on the stock prices of a green energy company. Epistemological, Phenomenological, Axiological and Ontological assumptions of Green Energy Management (GEM) Theory were utilized for selecting Air Products and Chemicals Inc. (APD) as the case study. Exxon Mobil (XOM) was used as a control for triangulation purposes. The period of time examined was between January of 1999 and December of 2008. Monthly stock prices for APD and XOM for the ten year period of time were collected from the New York Stock Exchange. Monthly U.S. crude oil spot prices for the ten year period of time were collected from the US Energy Information Administration. The data was entered into SPSS 17.0 software in order to conduct cross-correlation analysis. The six cross-correlation assumptions were satisfied in order to conduct a Cross-correlation Mirror Test (CCMT). The CCMT established the lag time direction and verified that U.S. crude oil spot prices serve as white noise for stock prices of APD and XOM. The Theory of Relative Weakness was employed in order to analyze the results. A 2 year period of time between December, 2006 and December, 2008 was examined. The correlation coefficient r = - .155 indicates that U.S. crude oil spot prices lead APD stock prices by 4 months. During the same 2 year period of time, U.S. crude oil spot prices lead XOM stock prices by 4 months at r = -.283. XOM stock prices and APD stock prices were positively correlated with 0 lag in time with a positive r = .566. The 4 month cycle was an exact match between APD stock prices, XOM stock prices and U.S. crude oil spot prices. The 4 month cycle was due to the random price fluctuation of U.S. crude oil spot prices that obscured the true stock prices of APD and XOM for the 2 year period of time.

  11. Simultaneous and rapid detection of white spot syndrome virus and yellow head virus infection in shrimp with a dual immunochromatographic strip test.

    PubMed

    Sithigorngul, Paisarn; Rukpratanporn, Sombat; Chaivisuthangkura, Parin; Sridulyakul, Pattarin; Longyant, Siwaporn

    2011-04-01

    A strip test for the dual detection of white spot syndrome virus (WSSV) and yellow head virus (YHV) was developed using monoclonal antibodies (MAbs) specific to the WSSV major envelope protein VP28 (W1 and W30) and the YHV nucleocapsid protein p20 (Y19 and Y21). The MAbs W30 and Y19 were conjugated with colloidal gold and sprayed onto a glass fiber pad that was placed adjacent to a sample chamber. The MAbs W1 and Y21 and the goat anti-mouse immunoglobulin G (GAM) antibody were sprayed onto a nitrocellulose membrane in strips at positions designated W, Y and C, respectively. These test strips were placed in plastic cases and stored desiccated in a plastic bag. The test strips were assessed for their ability to detect WSSV and YHV simultaneously using pleopods sampled from shrimp. A pleopod homogenate in application buffer 100μl was applied to the sample chamber to flow through the nitrocellulose membrane strip, and antibody-protein complexes could be observed within 15min. In sample from shrimp infected with WSSV and/or YHV, viral protein bound to the colloidal gold-conjugated MAbs. These complexes were captured by the MAbs at the W and/or Y test lines, resulting in the appearance of reddish-purple coloured bands. Any unbound colloidal gold-conjugated MAbs migrated pass the W and Y lines would be captured by the GAM antibody, forming a band at position C. When samples not containing WSSV and YHV proteins or containing viral proteins at below the detection limit of the test, only the band at position C was observed. The sensitivity of the test was comparable to dot blot tests using single MAbs, and ∼500-fold less sensitive than a 1-step PCR test for WSSV and 1000-fold less sensitive than an RT-PCR test for YHV. Despite this lower sensitivity, the dual strip test has advantages in speed and simplicity in not requiring sophisticated equipment or specialized skills. The ability to co-detect WSSV and YHV provides simultaneously cost savings.

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

    PubMed Central

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

    2016-01-01

    In previous work, small ubiquitin-like modifier (SUMO) in hemocytes of Chinese shrimp Fenneropenaeus chinensis was found to be up-regulated post-white spot syndrome virus (WSSV) infection using proteomic approach. However, the role of SUMO in viral infection is still unclear. In the present work, full length cDNAs of SUMO (FcSUMO) and SUMO-conjugating enzyme E2 UBC9 (FcUBC9) were cloned from F. chinensis using rapid amplification of cDNA ends approach. The open reading frame (ORF) of FcSUMO encoded a 93 amino acids peptide with the predicted molecular weight (M.W) of 10.55 kDa, and the UBC9 ORF encoded a 160 amino acids peptide with the predicted M.W of 18.35 kDa. By quantitative real-time RT-PCR, higher mRNA transcription levels of FcSUMO and FcUBC9 were detected in hemocytes and ovary of F. chinensis, and the two genes were significantly up-regulated post WSSV infection. Subsequently, the recombinant proteins of FcSUMO and FcUBC9 were expressed in Escherichia coli BL21 (DE3), and employed as immunogens for the production of polyclonal antibody (PAb). Indirect immunofluorescence assay revealed that the FcSUMO and UBC9 proteins were mainly located in the hemocytes nuclei. By western blotting, a 13.5 kDa protein and a 18.7 kDa protein in hemocytes were recognized by the PAb against SUMO or UBC9 respectively. Furthermore, gene silencing of FcSUMO and FcUBC9 were performed using RNA interference, and the results showed that the number of WSSV copies and the viral gene expressions were inhibited by knockdown of either SUMO or UBC9, and the mortalities of shrimp were also reduced. These results indicated that FcSUMO and FcUBC9 played important roles in WSSV infection. PMID:26927328

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

    PubMed

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

    2017-03-15

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

  14. Blue maize: morphology and starch synthase characterization of starch granule.

    PubMed

    Utrilla-Coello, Rubi G; Agama-Acevedo, Edith; de la Rosa, Ana Paulina Barba; Martinez-Salgado, Jose L; Rodriguez-Ambriz, Sandra L; Bello-Perez, Luis A

    2009-03-01

    The use of pigmented maize varieties has increased due to their high anthocyanins content, but very few studies are reported about the starch properties of these grains. The aim of this work was to isolate the starch granules from pigmented blue maize and carry out the morphological, physicochemical, and biochemical characterization studies. The proximate composition of starch granules showed high protein contents, after purification, the blue maize starch presented lower protein amount than starch from white maize (control). Although the purity of starch granules was increased, the damaged starch (determined for the Maltase cross absence) was also increased. Scanning electron microscopy showed the presence of some pores and channels in the blue maize starch. The electrophoretic protein profiles showed differences in the bands that correspond to the enzymes involved in the starch biosynthesis; these differences could explain the variation in morphological characteristics of blue maize starches against starch from white maize.

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

    PubMed

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

    2016-07-01

    days 15 and 30 post-feeding. In two white spot syndrome virus (WSSV) infections, the weight gain, survival, and WSSV copies within the gills of the probiotic-treated shrimp significantly differed (p < 0.05) from those of the control group. Relatively efficient protection was associated with probiotic feeding. Results suggested that Bacillus PC465 feeding improves the growth performance, survival, digestion, and nutrient absorption of L. vannamei. Probiotic treatment also enhances the microbial structures in the gut, promotes the immune status of shrimp, and provides protection against viral infection. The supplementation with 10(9) CFU g(-1) can also improve the growth and survival of L. vannamei.

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

    PubMed

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

    2016-02-01

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

  17. Reinventing MaizeGDB

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The Maize Database (MaizeDB) to the Maize Genetics and Genomics Database (MaizeGDB) turns 20 this year, and such a significant milestone must be celebrated! With the release of the B73 reference sequence and more sequenced genomes on the way, the maize community needs to address various opportunitie...

  18. A comparison of faecal microbial populations of South African Windsnyer-type indigenous pigs (SAWIPs) and Large White × Landrace (LW × LR) crosses fed diets containing ensiled maize cobs.

    PubMed

    Kanengoni, Arnold T; Chimonyo, Michael; Tasara, Taurai; Cormican, Paul; Chapwanya, Aspinas; Ndimba, Bongani K; Dzama, Kennedy

    2015-07-01

    Faecal microbial communities in South African Windsnyer-type indigenous pigs (SAWIPs) and Large White × Landrace (LW × LR) crosses were investigated using high-throughput sequencing of the 16S rDNA genes. The faecal microbial communities in LW × LR crosses and SAWIPs fed control (CON) and high maize cob (HMC) diets were evaluated through parallel sequencing of 16S rDNA genes. Butrivibrio, Faecalibacterium and Desulfovibrio, although present in LW × LR pigs, were absent from the SAWIP microbial community. Bacteroides, Succiniclasticum, Peptococcus and Akkermansia were found in SAWIPs but not in LW × LR crosses. The ratios of Bacteroidia to Clostridia on the CON and HMC diets were similar (0.37 versus 0.39) in SAWIPs but different (0.24 versus 0.1) in LW × LR crosses. The faecal microbial profiles determined were different between the LW × LR and SAWIP breeds but not between pigs fed the CON and HMC diets. The composition of faecal bacterial communities in SAWIPs was determined for the first time. The differences in microbial communities detected may explain the enhanced ability of SAWIPs to digest fibrous diets compared with the LW × LR crosses.

  19. SPOT Program

    NASA Technical Reports Server (NTRS)

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

    2010-01-01

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

  20. Dark Spots

    NASA Technical Reports Server (NTRS)

    2006-01-01

    Dark spots (left) and 'fans' appear to scribble dusty hieroglyphics on top of the Martian south polar cap in two high-resolution Mars Global Surveyor, Mars Orbiter Camera images taken in southern spring. Each image is about 3-kilometers wide (2-miles).

  1. Defrosting Spots

    NASA Technical Reports Server (NTRS)

    2005-01-01

    3 October 2005 This Mars Global Surveyor (MGS) Mars Orbiter Camera (MOC) image shows dark, defrosting spots formed on a polygon-cracked plain in the south polar region of Mars. The surface was covered with carbon dioxide frost during the previous winter. In spring, the material begins to sublime away, creating a pattern of dark spots that sometimes have wind streaks emanating from them, as wind carries away or erodes the frost.

    Location near: 87.2oS, 28.4oW Image width: width: 3 km (1.9 mi) Illumination from: upper left Season: Southern Spring

  2. The iojap gene in maize

    SciTech Connect

    Martienssen, Robert

    2001-12-01

    The classical maize mutant iojap (Iodent japonica) has variegated green and white leaves. Green sectors have cells with normal chloroplasts whereas white sectors have cells where plastids fail to differentiate. These mutant plastids, when transmitted through the female gametophyte, do not recover in the presence of wild type Iojap. We cloned the Ij locus, and we have investigated the mechanism of epigenetic inheritance and phenotypic expression. More recently, a modifier of this type of variegation, ''Inhibitor of striate'', has also been cloned. Both the iojap and inhibitor of striate proteins have homologs in bacteria and are members of ancient conserved families found in multiple species. These tools can be used to address fundamental questions of inheritance and variegation associated with this classical conundrum of maize genetics. Since the work of Rhoades there has been considerable speculation concerning the nature of the Iojap gene product, the origin of leaf variegation and the mechanism behind the material inheritance of defective plastids. This has made Iojap a textbook paradigm for cytoplasmic inheritance and nuclear-organellar interaction for almost 50 years. Cloning of the Iojap gene in maize, and homologs in other plants and bacteria, provides a new means to address the origin of heteroplastidity, variegation and cytoplasmic inheritance in higher plants.

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

    PubMed

    Kim, Hyoung Jun; Kwon, Se Ryun

    2014-12-01

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

  4. The morphology and genetic characterization of Iheringascaris goai n. sp. (Nematoda: Raphidascarididae) from the intestine of the silver whiting and spotted catfish off the central west coast of India.

    PubMed

    Malhotra, A; Jaiswal, N; Malakar, A K; Verma, M S; Singh, H R; Lakra, W S; Malhotra, S K; Shamsi, S

    2012-09-01

    In this study a new species of nematode, Iheringascaris goai n. sp., is reported from two fish hosts, including silver whiting, Sillago sihama, and spotted catfish, Arius maculatus, caught off the Central West Coast of India at Goa. The new species can be differentiated morphologically from I. inquies, the most closely related species collected from cohabiting marine fish. The distinguishing characteristics are distinct cuticular striations, a unilateral excretory system, the presence of dentigerous ridges on the inner margin of the lips and the ratio of oesophagus to body length. In males, the ratio of spicules to body length is higher and the number of pre-anal papillae is less in comparison to those in I. inquies. In addition, the tail curves ventrad in males, while in females, the vulva is post-equatorial. The sequence alignment of 18S rDNA and cytochrome c oxidase subunit I with sequences of known species selected from the same superfamily shows a significant difference. The morphological and molecular differences reported here can, therefore, be used to assign the specimen to a new species.

  5. The genetics of leaf flecking in maize and its relationship to plant defense and disease resistance

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Physiological/genetic leaf spotting, or flecking, is a mild lesion phenotype observed on the leaves of several commonly used maize inbred lines and has been anecdotally linked to enhanced broad-spectrum disease resistance. Flecking was assessed in the maize nested association mapping (NAM) populati...

  6. Evaluating the silencing suppressor activity of proteins encoded by maize rayado fino virus

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Maize rayado fino virus (MRFV), the type member of the genus Marafivirus, family Tymoviridae, is transmitted in a persistent, circulative manner by leafhoppers of the genus Dalbulus. Symptoms of MRFV infection on leaves of its maize host are small chlorotic spots that coalesce into short stripes. T...

  7. Mongolian spots.

    PubMed

    Gupta, Divya; Thappa, Devinder Mohan

    2013-01-01

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

  8. White shrimp Litopenaeus vannamei immersed in seawater containing Sargassum hemiphyllum var. chinense powder and its extract showed increased immunity and resistance against Vibrio alginolyticus and white spot syndrome virus.

    PubMed

    Huynh, Truong-Giang; Yeh, Su-Tuen; Lin, Yong-Chin; Shyu, Jeng-Feng; Chen, Li-Li; Chen, Jiann-Chu

    2011-08-01

    This study was to examine the immune response of white shrimp Litopenaeus vannamei and its resistance against Vibrio alginolyticus and WSSV when shrimp received the Sargassum hemiphyllum var. chinense powder and its hot-water extract. Both powder and extract showed activation of prophenoloxidase and generation of superoxide anion in the shrimp in vitro. The haemocyte count, phenoloxidase (PO) activity, respiratory burst, and lysozyme activity were examined after the shrimp were immersed in seawater containing S. hemiphyllum var. chinense powder or its extract at 0, 100, 300, and 500 mg L⁻¹ for 1, 3, and 5 h. These immune parameters of shrimp immersed in 300 and 500 mg L⁻¹ powder, and 100 and 300 mg L⁻¹ extract were significantly higher than those of control shrimp after 3 h, but slightly decreased after 5 h. In another experiment, shrimp immersed in seawater containing the powder or the extract at 0, 100, 300, and 500 mg L⁻¹ after 3 h were challenged with V. alginolyticus at 8 × 10⁵ colony-forming unit (cfu) shrimp⁻¹, or challenged with WSSV at 1 × 10⁵ copies shrimp⁻¹, and then placed in seawater. Survival rate of shrimp immersed in 500 mg L⁻¹ powder was significantly higher than that of control shrimp after 24-120 h in the V. alginolyticus-challenge test, and after 72 h in the WSSV-challenge test, respectively. Survival rate of shrimp immersed in 300 mg L⁻¹ extract was significantly higher than that of control shrimp after 72-120 h in both V. alginolyticus-challenge and WSSV-challenge tests. It was concluded that the shrimp immersed in seawater containing the powder at 500 mg L⁻¹, and the extract at 300 mg L⁻¹ had increased immunity and resistance against V. alginolyticus infection, and the shrimp that received extract at 300 mg L⁻¹ showed resistance against WSSV infection.

  9. Metabolizable energy, nitrogen balance, and ileal digestibility of amino acids in quality protein maize for pigs

    PubMed Central

    2014-01-01

    Background To compare the nutritional value and digestibility of five quality protein maize (QPM) hybrids to that of white and yellow maize, two experiments were carried out in growing pigs. In experiment 1, the energy metabolizability and the nitrogen balance of growing pigs fed one of five QPM hybrid diets were compared against those of pigs fed white or yellow maize. In experiment 2, the apparent and standardized ileal digestibility (AID and SID, respectively) of proteins and amino acids from the five QPM hybrids were compared against those obtained from pigs fed white and yellow maize. In both experiments, the comparisons were conducted using contrasts. Results The dry matter and nitrogen intakes were higher in the pigs fed the QPM hybrids (P < 0.05) than in the pigs fed white or yellow maize. Energy digestibility (P < 0.001) and metabolizability (P < 0.01) were higher in the pigs fed the white and yellow maize diets than in those fed the QPM diets. The AID of lysine was higher (P < 0.01) in the QPM diets than in the white and yellow maize. The AIDs of leucine, isoleucine, valine, phenylalanine, and methionine were lower in the QPM diets than those of maize (white and yellow) (all P < 0.05). Maize (white and yellow) had greater SIDs of leucine, isoleucine, valine, phenylalanine, glutamic acid, serine, alanine, tyrosine, and proline (P < 0.05). Conclusions Based on these results, it was concluded that QPM had a lower metabolizable energy content and a higher amount of digestible lysine than normal maize. PMID:25045520

  10. Multiplex real-time PCR and high-resolution melting analysis for detection of white spot syndrome virus, yellow-head virus, and Penaeus monodon densovirus in penaeid shrimp.

    PubMed

    Panichareon, Benjaporn; Khawsak, Paisarn; Deesukon, Warin; Sukhumsirichart, Wasana

    2011-12-01

    A multiplex real-time PCR and high-resolution melting (HRM) analysis was developed to detect simultaneously three of the major viruses of penaeid shrimp including white spot syndrome virus (WSSV), yellow-head virus (YHV), and Penaeus monodon densovirus (PmDNV). Plasmids containing DNA/cDNA fragments of WSSV and YHV, and genomic DNAs of PmDNV and normal shrimp were used to test sensitivity of the procedure. Without the need of any probe, the products were identified by HRM analysis after real-time PCR amplification using three sets of viral specific primers. The results showed DNA melting curves that were specific for individual virus. No positive result was detected with nucleic acids from shrimp, Penaeus monodon nucleopolyhedrovirus (PemoNPV), Penaeus stylirostris densovirus (PstDNV), or Taura syndrome virus (TSV). The detection limit for PmDNV, YHV and WSSV DNAs were 40fg, 50fg, and 500fg, respectively, which was 10 times more sensitive than multiplex real-time PCR analyzed by agarose gel electrophoresis. In viral nucleic acid mixtures, HRM analysis clearly identified each virus in dual and triple infection. To test the capability to use this method in field, forty-one of field samples were examined by HRM analysis in comparison with agarose gel electrophoresis. For HRM analysis, 11 (26.83%), 9 (21.95%), and 4 (9.76%) were infected with WSSV, PmDNV, and YHV, respectively. Agarose gel electrophoresis detected lesser number of PmDNV infection which may due to the limit of sensitivity. No multiple infection was found in these samples. This method provides a rapid, sensitive, specific, and simultaneous detection of three major viruses making it as a useful tool for diagnosis and epidemiological studies of these viruses in shrimp and carriers.

  11. 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.

    PubMed

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

    2013-02-01

    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.

  12. MaizeCyc: Metabolic networks in maize

    Technology Transfer Automated Retrieval System (TEKTRAN)

    MaizeCyc is a catalog of known and predicted metabolic and transport pathways that enables plant researchers to graphically represent the metabolome of maize (Zea mays), thereby supporting integrated systems-biology analysis. Supported analyses include molecular and genetic/phenotypic profiling (e.g...

  13. Southern Spots

    NASA Technical Reports Server (NTRS)

    2005-01-01

    [figure removed for brevity, see original site] Context image for PIA03092 Southern Spots

    This VIS image of the south polar region was collected during the summer season. The markings of the pole are very diverse and easy to see after the winter frost has been removed.

    Image information: VIS instrument. Latitude 79.7S, Longitude 56.6E. 17 meter/pixel resolution.

    Note: this THEMIS visual image has not been radiometrically nor geometrically calibrated for this preliminary release. An empirical correction has been performed to remove instrumental effects. A linear shift has been applied in the cross-track and down-track direction to approximate spacecraft and planetary motion. Fully calibrated and geometrically projected images will be released through the Planetary Data System in accordance with Project policies at a later time.

    NASA's Jet Propulsion Laboratory manages the 2001 Mars Odyssey mission for NASA's Office of Space Science, Washington, D.C. The Thermal Emission Imaging System (THEMIS) was developed by Arizona State University, Tempe, in collaboration with Raytheon Santa Barbara Remote Sensing. The THEMIS investigation is led by Dr. Philip Christensen at Arizona State University. Lockheed Martin Astronautics, Denver, is the prime contractor for the Odyssey project, and developed and built the orbiter. Mission operations are conducted jointly from Lockheed Martin and from JPL, a division of the California Institute of Technology in Pasadena.

  14. Bacterial Diversity and Mycotoxin Reduction During Maize Fermentation (Steeping) for Ogi Production

    PubMed Central

    Okeke, Chiamaka A.; Ezekiel, Chibundu N.; Nwangburuka, Cyril C.; Sulyok, Michael; Ezeamagu, Cajethan O.; Adeleke, Rasheed A.; Dike, Stanley K.; Krska, Rudolf

    2015-01-01

    Bacterial diversity and community structure of two maize varieties (white and yellow) during fermentation/steeping for ogi production, and the influence of spontaneous fermentation on mycotoxin reduction in the gruel were studied. A total of 142 bacterial isolates obtained at 24–96 h intervals were preliminarily identified by conventional microbiological methods while 60 selected isolates were clustered into 39 OTUs consisting of 15 species, 10 genera, and 3 phyla by 16S rRNA sequence analysis. Lactic acid bacteria constituted about 63% of all isolated bacteria and the genus Pediococcus dominated (white maize = 84.8%; yellow maize = 74.4%). Pediococcus acidilactici and Lactobacillus paraplantarum were found at all steeping intervals of white and yellow maize, respectively, while P. claussenii was present only at the climax stage of steeping white maize. In both maize varieties, P. pentosaceus was found at 24–72 h. Mycotoxin concentrations (μg/kg) in the unsteeped grains were: white maize (aflatoxin B1 = 0.60; citrinin = 85.8; cyclopiazonic acid = 23.5; fumonisins (B1/B2/B3) = 68.4–483; zearalenone = 3.3) and yellow maize (aflatoxins (B1/B2/M1) = 22.7–513; citrinin = 16,800; cyclopiazonic acid = 247; fumonisins (B1/B2/B3) = 252–1,586; zearalenone = 205). Mycotoxins in both maize varieties were significantly (p < 0.05) reduced across steeping periods. This study reports for the first time: (a) the association of L. paraplantarum, P. acidilactici, and P. claussenii with ogi production from maize, (b) citrinin occurrence in Nigerian maize and ogi, and (c) aflatoxin M1, citrinin and cyclopiazonic acid degradation/loss due to fermentation in traditional cereal-based fermented food. PMID:26697001

  15. Are 'hot spots' hot spots?

    NASA Astrophysics Data System (ADS)

    Foulger, Gillian R.

    2012-07-01

    The term 'hot spot' emerged in the 1960s from speculations that Hawaii might have its origins in an unusually hot source region in the mantle. It subsequently became widely used to refer to volcanic regions considered to be anomalous in the then-new plate tectonic paradigm. It carried with it the implication that volcanism (a) is emplaced by a single, spatially restricted, mongenetic melt-delivery system, assumed to be a mantle plume, and (b) that the source is unusually hot. This model has tended to be assumed a priori to be correct. Nevertheless, there are many geological ways of testing it, and a great deal of work has recently been done to do so. Two fundamental problems challenge this work. First is the difficulty of deciding a 'normal' mantle temperature against which to compare estimates. This is usually taken to be the source temperature of mid-ocean ridge basalts (MORBs). However, Earth's surface conduction layer is ˜200 km thick, and such a norm is not appropriate if the lavas under investigation formed deeper than the 40-50 km source depth of MORB. Second, methods for estimating temperature suffer from ambiguity of interpretation with composition and partial melt, controversy regarding how they should be applied, lack of repeatability between studies using the same data, and insufficient precision to detect the 200-300 °C temperature variations postulated. Available methods include multiple seismological and petrological approaches, modelling bathymetry and topography, and measuring heat flow. Investigations have been carried out in many areas postulated to represent either (hot) plume heads or (hotter) tails. These include sections of the mid-ocean spreading ridge postulated to include ridge-centred plumes, the North Atlantic Igneous Province, Iceland, Hawaii, oceanic plateaus, and high-standing continental areas such as the Hoggar swell. Most volcanic regions that may reasonably be considered anomalous in the simple plate-tectonic paradigm have been

  16. Red Spot Movie

    NASA Technical Reports Server (NTRS)

    2000-01-01

    This brief movie shows counterclockwise atmospheric motion around Jupiter's Great Red Spot. The clip was made from blue-filter images taken with the narrow-angle camera on NASA's Cassini spacecraft during seven separate rotations of Jupiter between Oct. 1 and Oct. 5, 2000.

    The clip also shows the eastward and westward motion of the zonal jets, seen as the horizontal stripes flowing in opposite directions. The zonal jets circle the planet. As far as can be determined from both Earth-based and spacecraft measurements, the positions and speeds of the jets have not changed for 100 years. Since Jupiter is a fluid planet without a solid boundary, the jet speeds are measured relative to Jupiter's magnetic field, which rotates, wobbling like a top because of its tilt, every 9 hours 55.5 minutes. The movie shows motions in the magnetic reference frame, so winds to the west correspond to features that are rotating a little slower than the magnetic field, and eastward winds correspond to features rotating a little faster.

    Because the Red Spot is in the southern hemisphere, the direction of motion indicates it is a high-pressure center. Small bright clouds appear suddenly to the west of the Great Red Spot. Scientists suspect these small white features are lightning storms. The storms eventually merge with the Red Spot and surrounding jets, and may be the main energy source for the large-scale features.

    The smallest features in the movie are about 500 kilometers (about 300 miles) across. The spacing of the movie frames in time is not uniform; some consecutive images are separated by two Jupiter rotations, and some by one. The images have been re-projected using a simple cylindrical map projection. They show an area from 50 degrees north of Jupiter's equator to 50 degrees south, extending 100 degrees east-west, about one quarter of Jupiter's circumference.

    Cassini is a cooperative project of NASA, the European Space Agency and the Italian Space Agency. The Jet

  17. Jovian Dark Spot

    NASA Technical Reports Server (NTRS)

    1998-01-01

    A recently discovered black spot in Jupiter's clouds is darker than any feature ever before observed on the giant planet. The spot may be the result of a downward spiraling wind that blows away high clouds and reveals deeper, very dark cloud layers. These three panels depict the same area of Jupiter's atmosphere. A map of Jovian temperatures near 250 millibar pressure (top) panel is derived from the photopolarimeter-radiometer instrument on NASA's Galileo Jupiter orbiter. This map is compared with maps derived from images of the same area in visible light (middle panel)and thermal radiation sensitive to cloud-top temperatures (bottom panel).

    The single downward-pointing arrow in the top panel indicates the location of a warm area that corresponds to the position of a so-called 'black spot'(shown in the middle panel), a feature that is about a year old. Features this dark are rare on Jupiter. The bottom panel, sensitive to temperatures at Jupiter's cloud tops, shows this feature as a bright object, meaning that upper-level cold clouds are missing - allowing us to see deeper into Jupiter's warmer interior. The dark visible appearance of the feature than most likely represents the color of very deep clouds. The warm temperatures and cloud-free conditions imply that this feature is a region where dry upper-atmospheric gas is being forced to converge, is warmed up and then forced to descend, clearing out clouds. It is the opposite of wet, upwelling gas in areas such as Jupiter's Great Red Spot or white ovals. On the other hand, it is unlike the dry and relatively cloudless feature into which the Galileo probe descended in 1995, because that region had the same temperatures as its surroundings and did not appear nearly as dark as this new spot.

    The temperatures sampled by the photopolarimeter radiometer are near the top of Jupiter's troposphere, where wind motions control the atmosphere. The top row of arrows shows the location of temperature waves in a warm region

  18. The TIGR Maize Database.

    PubMed

    Chan, Agnes P; Pertea, Geo; Cheung, Foo; Lee, Dan; Zheng, Li; Whitelaw, Cathy; Pontaroli, Ana C; SanMiguel, Phillip; Yuan, Yinan; Bennetzen, Jeffrey; Barbazuk, William Brad; Quackenbush, John; Rabinowicz, Pablo D

    2006-01-01

    Maize is a staple crop of the grass family and also an excellent model for plant genetics. Owing to the large size and repetitiveness of its genome, we previously investigated two approaches to accelerate gene discovery and genome analysis in maize: methylation filtration and high C(0)t selection. These techniques allow the construction of gene-enriched genomic libraries by minimizing repeat sequences due to either their methylation status or their copy number, yielding a 7-fold enrichment in genic sequences relative to a random genomic library. Approximately 900,000 gene-enriched reads from maize were generated and clustered into Assembled Zea mays (AZM) sequences. Here we report the current AZM release, which consists of approximately 298 Mb representing 243,807 sequence assemblies and singletons. In order to provide a repository of publicly available maize genomic sequences, we have created the TIGR Maize Database (http://maize.tigr.org). In this resource, we have assembled and annotated the AZMs and used available sequenced markers to anchor AZMs to maize chromosomes. We have constructed a maize repeat database and generated draft sequence assemblies of 287 maize bacterial artificial chromosome (BAC) clone sequences, which we annotated along with 172 additional publicly available BAC clones. All sequences, assemblies and annotations are available at the project website via web interfaces and FTP downloads.

  19. MaizeGDB, the maize model organism database

    Technology Transfer Automated Retrieval System (TEKTRAN)

    MaizeGDB is the maize research community's database for maize genetic and genomic information. In this seminar I will outline our current endeavors including a full website redesign, the status of maize genome assembly and annotation projects, and work toward genome functional annotation. Mechanis...

  20. Sorghum and Maize

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Sorghum and maize are closely related cereal grains grown throughout the world. Sorghum, a drought tolerant crop grown in semi-arid regions, is a basic food staple in many parts of the developing world, while primarily an animal feed in western countries. Maize, a major worldwide crop, is used for...

  1. Maize Genetic Resources

    Technology Transfer Automated Retrieval System (TEKTRAN)

    This chapter describes the resources held at the Maize Genetics Cooperation • Stock Center in detail and also provides some information about the North Central Regional Plant Introduction Station (NCRPIS) in Ames, IA, Centro Internacional de Mejoramiento de Maiz y Trigo (CIMMYT) in Mexico, and the N...

  2. MAIZE ALLELIC DIVERSITY PROJECT

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Of the estimated 250-300 races of maize, only 24 races are represented in materials utilized by the Germplasm Enhancement of Maize (GEM) project, a collaborative effort between USDA-ARS and public and private sector research scientists. This is largely a result of poor performance of many races in ...

  3. Mongolian blue spots (image)

    MedlinePlus

    Mongolian blue spots are flat bluish- to bluish-gray skin markings commonly appearing at birth or shortly ... back and also can appear on the shoulders. Mongolian spots are benign and are not associated with ...

  4. Rocky Mountain spotted fever

    MedlinePlus

    ... this page: //medlineplus.gov/ency/article/000654.htm Rocky Mountain spotted fever To use the sharing features on this page, please enable JavaScript. Rocky Mountain spotted fever is a disease caused by a type of ...

  5. Proteomic analysis reveals differential accumulation of small heat shock proteins and late embryogenesis abundant proteins between ABA-deficient mutant vp5 seeds and wild-type Vp5 seeds in maize

    PubMed Central

    Wu, Xiaolin; Gong, Fangping; Yang, Le; Hu, Xiuli; Tai, Fuju; Wang, Wei

    2014-01-01

    ABA is a major plant hormone that plays important roles during many phases of plant life cycle, including seed development, maturity and dormancy, and especially the acquisition of desiccation tolerance. Understanding of the molecular basis of ABA-mediated plant response to stress is of interest not only in basic research on plant adaptation but also in applied research on plant productivity. Maize mutant viviparous-5 (vp5), deficient in ABA biosynthesis in seeds, is a useful material for studying ABA-mediated response in maize. Due to carotenoid deficiency, vp5 endosperm is white, compared to yellow Vp5 endosperm. However, the background difference at proteome level between vp5 and Vp5 seeds is unclear. This study aimed to characterize proteome alterations of maize vp5 seeds and to identify ABA-dependent proteins during seed maturation. We compared the embryo and endosperm proteomes of vp5 and Vp5 seeds by gel-based proteomics. Up to 46 protein spots, most in embryos, were found to be differentially accumulated between vp5 and Vp5. The identified proteins included small heat shock proteins (sHSPs), late embryogenesis abundant (LEA) proteins, stress proteins, storage proteins and enzymes among others. However, EMB564, the most abundant LEA protein in maize embryo, accumulated in comparable levels between vp5 and Vp5 embryos, which contrasted to previously characterized, greatly lowered expression of emb564 mRNA in vp5 embryos. Moreover, LEA proteins and sHSPs displayed differential accumulations in vp5 embryos: six out of eight identified LEA proteins decreased while nine sHSPs increased in abundance. Finally, we discussed the possible causes of global proteome alterations, especially the observed differential accumulation of identified LEA proteins and sHSPs in vp5 embryos. The data derived from this study provides new insight into ABA-dependent proteins and ABA-mediated response during maize seed maturation. PMID:25653661

  6. Pollen-Mediated Gene Flow in Maize: Implications for Isolation Requirements and Coexistence in Mexico, the Center of Origin of Maize

    PubMed Central

    Gutiérrez, Marco Antonio; Guzmán Rodríguez, José Luis; Horak, Michael J.; Madueño Martínez, Jesús Ignacio; Schapaugh, Adam W.; Stojšin, Duška; Uribe Montes, Hugo Raúl

    2015-01-01

    Mexico, the center of origin of maize (Zea mays L.), has taken actions to preserve the identity and diversity of maize landraces and wild relatives. Historically, spatial isolation has been used in seed production to maintain seed purity. Spatial isolation can also be a key component for a strategy to minimize pollen-mediated gene flow in Mexico between transgenic maize and sexually compatible plants of maize conventional hybrids, landraces, and wild relatives. The objective of this research was to generate field maize-to-maize outcrossing data to help guide coexistence discussions in Mexico. In this study, outcrossing rates were determined and modeled from eight locations in six northern states, which represent the most economically important areas for the cultivation of hybrid maize in Mexico. At each site, pollen source plots were planted with a yellow-kernel maize hybrid and surrounded by plots with a white-kernel conventional maize hybrid (pollen recipient) of the same maturity. Outcrossing rates were then quantified by assessing the number of yellow kernels harvested from white-kernel hybrid plots. The highest outcrossing values were observed near the pollen source (12.9% at 1 m distance). The outcrossing levels declined sharply to 4.6, 2.7, 1.4, 1.0, 0.9, 0.5, and 0.5% as the distance from the pollen source increased to 2, 4, 8, 12, 16, 20, and 25 m, respectively. At distances beyond 20 m outcrossing values at all locations were below 1%. These trends are consistent with studies conducted in other world regions. The results suggest that coexistence measures that have been implemented in other geographies, such as spatial isolation, would be successful in Mexico to minimize transgenic maize pollen flow to conventional maize hybrids, landraces and wild relatives. PMID:26162097

  7. Pollen-Mediated Gene Flow in Maize: Implications for Isolation Requirements and Coexistence in Mexico, the Center of Origin of Maize.

    PubMed

    Baltazar, Baltazar M; Castro Espinoza, Luciano; Espinoza Banda, Armando; de la Fuente Martínez, Juan Manuel; Garzón Tiznado, José Antonio; González García, Juvencio; Gutiérrez, Marco Antonio; Guzmán Rodríguez, José Luis; Heredia Díaz, Oscar; Horak, Michael J; Madueño Martínez, Jesús Ignacio; Schapaugh, Adam W; Stojšin, Duška; Uribe Montes, Hugo Raúl; Zavala García, Francisco

    2015-01-01

    Mexico, the center of origin of maize (Zea mays L.), has taken actions to preserve the identity and diversity of maize landraces and wild relatives. Historically, spatial isolation has been used in seed production to maintain seed purity. Spatial isolation can also be a key component for a strategy to minimize pollen-mediated gene flow in Mexico between transgenic maize and sexually compatible plants of maize conventional hybrids, landraces, and wild relatives. The objective of this research was to generate field maize-to-maize outcrossing data to help guide coexistence discussions in Mexico. In this study, outcrossing rates were determined and modeled from eight locations in six northern states, which represent the most economically important areas for the cultivation of hybrid maize in Mexico. At each site, pollen source plots were planted with a yellow-kernel maize hybrid and surrounded by plots with a white-kernel conventional maize hybrid (pollen recipient) of the same maturity. Outcrossing rates were then quantified by assessing the number of yellow kernels harvested from white-kernel hybrid plots. The highest outcrossing values were observed near the pollen source (12.9% at 1 m distance). The outcrossing levels declined sharply to 4.6, 2.7, 1.4, 1.0, 0.9, 0.5, and 0.5% as the distance from the pollen source increased to 2, 4, 8, 12, 16, 20, and 25 m, respectively. At distances beyond 20 m outcrossing values at all locations were below 1%. These trends are consistent with studies conducted in other world regions. The results suggest that coexistence measures that have been implemented in other geographies, such as spatial isolation, would be successful in Mexico to minimize transgenic maize pollen flow to conventional maize hybrids, landraces and wild relatives.

  8. Homogentisate solanesyl transferase (HST) cDNA’s in maize

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Maize white seedling 3 (w3) has served as a model albino-seedling mutant since its discovery in 1923. We show that the w3 phenotype is caused by disruptions in homogentisate solanesyl transferase (HST), an enzyme that catalyzes the committed step in plastoquinone-9 (PQ9) biosynthesis. This reaction ...

  9. Photomorphogenic Responses in Maize Seedling Development1[w

    PubMed Central

    Markelz, Nicole H.; Costich, Denise E.; Brutnell, Thomas P.

    2003-01-01

    As an emerging maize (Zea mays) seedling senses light, there is a decrease in the rate of mesocotyl elongation, an induction of root growth, and an expansion of leaves. In leaf tissues, mesophyll and bundle sheath cell fate is determined, and the proplastids of each differentiate into the dimorphic chloroplasts typical of each cell type. Although it has been inferred from recent studies in several model plant species that multiple photoreceptor systems mediate this process, surprisingly little is known of light signal transduction in maize. Here, we examine two photomorphogenic responses in maize: inhibition of mesocotyl elongation and C4 photosynthetic differentiation. Through an extensive survey of white, red, far-red, and blue light responses among a diverse collection of germplasm, including a phytochrome-deficient mutant elm1, we show that light response is a highly variable trait in maize. Although all inbreds examined appear to have a functional phytochrome signal transduction pathway, several lines showed reduced sensitivity to blue light. A significant correlation was observed between light response and subpopulation, suggesting that light responsiveness may be a target of artificial selection. An examination of C4 gene expression patterns under various light regimes in the standard W22 inbred and elm1 indicate that cell-specific patterns of C4 gene expression are maintained in fully differentiated tissues independent of light quality. To our knowledge, these findings represent the first comprehensive survey of light response in maize and are discussed in relation to maize breeding strategies. PMID:14645729

  10. Maize (Zea mays L.).

    PubMed

    Frame, Bronwyn; Warnberg, Katey; Main, Marcy; Wang, Kan

    2015-01-01

    Agrobacterium tumefaciens-mediated transformation is an effective method for introducing genes into maize. In this chapter, we describe a detailed protocol for genetic transformation of the maize genotype Hi II. Our starting plant material is immature embryos cocultivated with an Agrobacterium strain carrying a standard binary vector. In addition to step-by-step laboratory transformation procedures, we include extensive details in growing donor plants and caring for transgenic plants in the greenhouse.

  11. The Earth's Hot Spots.

    ERIC Educational Resources Information Center

    Vink, Gregory E.; And Others

    1985-01-01

    Hot spots are isolated areas of geologic activity where volcanic eruptions, earthquakes, and upwelling currents occur far from plate boundaries. These mantle plumes are relatively stable and crustal plates drift over them. The nature and location of hot spots (with particular attention to the Hawaiian Islands and Iceland) are discussed. (DH)

  12. Still from Red Spot Movie

    NASA Technical Reports Server (NTRS)

    2000-01-01

    This image is one of seven from the narrow-angle camera on NASA's Cassini spacecraft assembled as a brief movie of cloud movements on Jupiter. It was taken with a blue filter. The smallest features visible are about 500 kilometers (about 300 miles) across.

    Small bright clouds appear suddenly to the west of the Great Red Spot. Based on data from NASA's Galileo spacecraft, scientists suspect that these small white features are lightning storms, where falling raindrops create an electrical charge. The lightning storms eventually merge with the Red Spot and surrounding jets, and may be the main energy source for these large-scale features. Imaging observations of the darkside of the planet in the weeks following Cassini's closest approach to Jupiter on Dec. 30, 2000 will search for lightning storms like these.

    This image was re-projected by cylindrical-map projection of an image taken in the first week of October 2000. It shows an area from 50 degrees north of Jupiter's equator to 50 degrees south, extending 100 degrees east west, about one quarter of Jupiter's circumference.

    Cassini is a cooperative project of NASA, the European Space Agency and the Italian Space Agency. The Jet Propulsion Laboratory, a division of the California Institute of Technology in Pasadena, manages the Cassini mission for NASA's Office of Space Science, Washington, D.C.

  13. Watermarking spot colors

    NASA Astrophysics Data System (ADS)

    Alattar, Osama M.; Reed, Alastair M.

    2003-06-01

    Watermarking of printed materials has usually focused on process inks of cyan, magenta, yellow and black (CMYK). In packaging, almost three out of four printed materials include spot colors. Spot colors are special premixed inks, which can be produced in a vibrant range of colors, often outside the CMYK color gamut. In embedding a watermark into printed material, a common approach is to modify the luminance value of each pixel in the image. In the case of process color work pieces, the luminance change can be scaled to the C, M, Y and K channels using a weighting function, to produce the desired change in luminance. In the case of spot color art designs, there is only one channel available and the luminance change is applied to this channel. In this paper we develop a weighting function to embed the watermark signal across the range of different spot colors. This weighting function normalizes visibility effect and signal robustness across a wide range of different spot colors. It normalizes the signal robustness level over the range of an individual spot color"s intensity levels. Further, it takes into account the sensitivity of the capturing device to the different spot colors.

  14. Genomic variation in maize

    SciTech Connect

    Rivin, C.J.

    1990-01-01

    We have endeavored to learn to learn how different DNA sequences and sequence arrangements contribute to genome plasticity in maize. We describe quantitative variation among maize inbred lines for tandemly arrayed and dispersed repeated DNA sequences and gene families, and qualitative variation for sequences homologous to the Mutator family of transposons. The potential of these sequences to undergo unequal crossing over, non-allelic (ectopic) recombination and transposition makes them a source of genome instability. We have found examples of rapid genomic change involving these sequences in F1 hybrids, tissue culture cells and regenerated plants.

  15. MaizeGDB - Past, present, and future

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The Maize Genetics and Genomics Database (MaizeGDB) turns 20 this year. This editorial outlines MaizeGDB's history and connection to the Maize Genetics Cooperation, describes key components of how the MaizeGDB interface will be completely redesigned over the course of the next two years to meet cur...

  16. Mononucleosis spot test

    MedlinePlus

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

  17. Lincoln's Spot Resolutions.

    ERIC Educational Resources Information Center

    Mueller, Jean West; Schamel, Wynell Burroughs

    1988-01-01

    Examines the events leading to and immediately following the declaration of war on Mexico in 1846. Includes the second and third pages of Abraham Lincoln's "Spot Resolutions" and presents teaching suggestions for interpreting the document and assessing public opinion. (GEA)

  18. SPOT4 Management Centre

    NASA Technical Reports Server (NTRS)

    Labrune, Yves; Labbe, X.; Roussel, A.; Vielcanet, P.

    1994-01-01

    In the context of the CNES SPOT4 program CISI is particularly responsible for the development of the SPOT4 Management Centre, part of the SPOT4 ground control system located at CNES Toulouse (France) designed to provide simultaneous control over two satellites. The main operational activities are timed to synchronize with satellite visibilities (ten usable passes per day). The automatic capability of this system is achieved through agenda services (sequence of operations as defined and planned by operator). Therefore, the SPOT4 Management Centre offers limited, efficient and secure human interventions for supervision and decision making. This paper emphasizes the main system characteristics as degree of automation, level of dependability and system parameterization.

  19. Scientists Spot 'Teetotaler' Gene

    MedlinePlus

    ... gov/news/fullstory_162265.html Scientists Spot 'Teetotaler' Gene Discovery might one day lead to drugs to ... HealthDay News) -- Scientists say they've identified a gene variant that dampens the desire to drink alcohol. ...

  20. (Genomic variation in maize)

    SciTech Connect

    Rivin, C.J.

    1991-01-01

    These studies have sought to learn how different DNA sequences and sequence arrangements contribute to genome plasticity in maize. We describe quantitative variation among maize inbred lines for tandemly arrayed and dispersed repeated DNA sequences and gene families, and qualitative variation for sequences homologous to the Mutator family of transposons. The potential of these sequences to undergo unequal crossing over, non-allelic (ectopic) recombination and transposition makes them a source of genome instability. We have found examples of rapid genomic change involving these sequences in Fl hybrids, tissue culture cells and regenerated plants. We describe the repetitive portion of the maize genome as composed primarily of sequences that vary markedly in copy number among different genetic stocks. The most highly variable is the 185 bp repeat associated with the heterochromatic chromosome knobs. Even in lines without visible knobs, there is a considerable quantity of tandemly arrayed repeats. We also found a high degree of variability for the tandemly arrayed 5S and ribosomal DNA repeats. While such variation might be expected as the result of unequal cross-over, we were surprised to find considerable variation among lower copy number, dispersed repeats as well. One highly repeated sequence that showed a complex tandem and dispersed arrangement stood out as showing no detectable variability among the maize lines. In striking contrast to the variability seen between the inbred stocks, individuals within a stock were indistinguishable with regard to their repeated sequence multiplicities.

  1. Betaine deficiency in maize

    SciTech Connect

    Lerma, C. ); Rich, P.J.; Ju, G.C.; Yang, Wenju; Rhodes, D. ); Hanson, A.D. )

    1991-04-01

    Maize (Zea mays L.) is a betaine-accumulating species, but certain maize genotypes lack betaine almost completely; a single recessive gene has been implicated as the cause of this deficiency. This study was undertaken to determine whether betaine deficiency in diverse maize germplasm is conditioned by the same genetic locus, and to define the biochemical lesion(s) involved. Complementation tests indicated that all 13 deficient genotypes tested shared a common locus. One maize population (P77) was found to be segregating for betaine deficiency, and true breeding individuals were used to produce related lines with and without betaine. Leaf tissue of both betaine-positive and betaine-deficient lines readily converted supplied betaine aldehyde to betaine, but only the betaine-containing line was able to oxidize supplied choline to betaine. This locates the lesion in betaine-deficient plants at the choline {r arrow} betaine aldehyde step of betaine synthesis. Consistent with this location, betaine-deficient plants were shown to have no detectable endogenous pool of betaine aldehyde.

  2. Monitoring of Maize Damage Caused by Western Corn Rootworm by Remote Sensing

    NASA Astrophysics Data System (ADS)

    Nádor, G.; Fényes, D.; Vasas, L.; Surek, G.

    2009-04-01

    The gradual dispersion of western corn rootworm (WCR) is becoming a serious maize pest in Europe, and all over the world. In 2008 using remote sensing data, the Remote Sensing Centre of Institute of Geodesy, Cartography and Remote Sensing (FÖMI RSC) carried out this project to identify WCR larval damage. Our goal with the present project is to assess and identify the disorder and structural changes caused by WCR larvae using optical (IRS-P6 AWiFS, IRS-P6 LISS, SPOT4 and SPOT5) and polarimetic radar (ALOS PALSAR) satellite images. We used 3 different individual features (Mono-maize feature, Optical feature, Radar feature) derived from remote sensing data to accomplish this goal. Findings were tested against on-the-spot ground assessments. Using radar polarimetry increased the accuracy significantly. The final results have implications for plant protection strategy, farming practices, pesticide producers, state authorities and research institutes.

  3. White House

    MedlinePlus

    ... to main content Jump to navigation the WHITE HOUSE President Donald J. Trump Get in Touch Home ... News Read the latest news from the White House Video Gallery View the most recent videos from ...

  4. Physicochemical, rheological and structural characteristics of starch in maize tortillas.

    PubMed

    Hernández-Uribe, Juan P; Ramos-López, Gonzalo; Yee-Madeira, Hernani; Bello-Pérez, Luis A

    2010-06-01

    Fresh and stored maize (white and blue) tortillas were evaluated for physicochemical, rheological and structural characteristics assessed by calorimetry, x-ray diffraction, Fourier transform infrared (FTIR) spectroscopy, dynamic viscoelastic tests, and high-performance size-exclusion chromatography. Two endotherms were found in studies of fresh and stored tortillas. The low temperature endotherm (50-56 degrees C) was due to reorganized (retrograded) amylopectin, while the high temperature endotherm (105-123 degrees C) was attributed to retrograded amylose. The enthalpy value for the lower temperature transition was minor than that of the high temperature transition. Fresh tortillas showed an amorphous starch arrangement by x-ray diffraction study. Stored samples showed the presence of peaks at 2theta = 17 masculine and 23 masculine, indicating re-crystallization of starch components. FTIR results confirmed the development of higher levels of starch crystals during storage. Differences in the viscoelastic parameters were also observed between fresh and stored samples. At the longest storage times, white tortillas were more rigid than blue tortillas. Molar mass values for starch increased for both white and blue tortillas as storage time progressed, though relatively higher values were obtained for white tortillas. More starch reorganization occurred in white tortillas, in accordance to calorimetric, x-ray diffraction, FTIR and rheological results. These results corroborate that changes occurring in tortillas during storage are related to reorganization of starch components, and the maize variety more than the color plays an important role.

  5. IR Spot Weld Inspect

    SciTech Connect

    Chen, Jian; Feng, Zhili

    2014-01-01

    In automotive industry, destructive inspection of spot welds is still the mandatory quality assurance method due to the lack of efficient non-destructive evaluation (NDE) tools. However, it is costly and time-consuming. Recently at ORNL, a new NDE prototype system for spot weld inspection using infrared (IR) thermography has been developed to address this problem. This software contains all the key functions that ensure the NDE system to work properly: system input/output control, image acquisition, data analysis, weld quality database generation and weld quality prediction, etc.

  6. MaizeGDB: The Maize Genetics and Genomics Database.

    Technology Transfer Automated Retrieval System (TEKTRAN)

    MaizeGDB is the community database for biological information about the crop plant Zea mays. Genomic, genetic, sequence, gene product, functional characterization, literature reference, and person/organization contact information are among the datatypes stored at MaizeGDB. At the project’s website...

  7. Genetic mechanisms of Maize dwarf mosaic virus resistance in maize

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Maize resistance to viruses has been well-characterized at the genetic level, and loci responsible for resistance to potyviruses including Maize dwarf mosaic virus (MDMV), Sugarcane mosaic virus (SCMV), Sorghum mosaic virus (SrMV), and Johnsongrass mosaic virus (JGMV), have been mapped in several ge...

  8. Rolling Spot Welder

    NASA Technical Reports Server (NTRS)

    Wagner, Garret E.; Fonteyne, Steve L.

    1990-01-01

    Wheeled tool speeds tack-welding operations. Spotwelds foil to parts in preparation for brazing. Includes electrode wheel rolling across foil. Welding current in electrode pulsed as electrode moves along, making series of uniformly-spaced low-current spot welds.

  9. Bacterial leaf spot

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Bacterial leaf spot has been reported in Australia (Queensland), Egypt, El Salvador, India, Japan, Nicaragua, Sudan, and the United States (Florida, Iowa, Kansas, Maryland, and Wisconsin). It occasionally causes locally severe defoliation and post-emergence damping-off and stunting. The disease is...

  10. Rocky Mountain Spotted Fever.

    PubMed

    Phillips, Jennan

    2017-01-01

    The tick-borne disease Rocky Mountain spotted fever (RMSF) can have deadly outcomes unless treated appropriately, yet nonspecific flu-like symptoms complicate diagnosis. Occupational health nurses must have a high index of suspicion with symptomatic workers and recognize that recent recreational or occupational activities with potential tick exposure may suggest RMSF.

  11. Arc spot grouping: An entanglement of arc spot cells

    SciTech Connect

    Kajita, Shin; Hwangbo, Dogyun; Ohno, Noriyasu; Tsventoukh, Mikhail M.; Barengolts, Sergey A.

    2014-12-21

    In recent experiments, clear transitions in velocity and trail width of an arc spot initiated on nanostructured tungsten were observed on the boundary of the thick and thin nanostructured layer regions. The velocity of arc spot was significantly decreased on the thick nanostructured region. It was suggested that the grouping decreased the velocity of arc spot. In this study, we try to explain the phenomena using a simple random walk model that has properties of directionality and self-avoidance. And grouping feature was added by installing an attractive force between spot cells with dealing with multi-spots. It was revealed that an entanglement of arc spot cells decreased the spot velocity, and spot cells tend to stamp at the same location many times.

  12. TV spots' impact.

    PubMed

    El-bakly, S

    1994-09-01

    The Information, Education and Communication (IEC) Center of the State Information Service was established in 1979 for the purpose of providing information to the people on the population issue. The Ministry of Information has accorded the State Information Service free TV and radio air time for family planning dramas and spots. In the early years information campaigns were organized to make people aware of the population problem by slogans, songs, and cartoons. Around 1984 misconceptions about family planning and contraceptives were attacked through a number of TV and radio spots. A few years later 21 spots on specific contraceptive methods were broadcast which were aired for three years over 3000 times. They were extremely successful. The impact of these TV spots was one of the major reasons why the contraceptive prevalence rate increased from 30% in 1984 to 38% in 1988 and 47% in 1992. Spots were also broadcast about the social implications of large families. The TV soap opera "And The Nile Flows On", with the family planning message interwoven into it, was very well received by the target audience. A program entitled "Wedding of the Month" features couples who know family planning well. The most successful radio program is a 15-20 minute long quiz show for residents of the villages where the Select Villages Project is being implemented. The State Information Service has 60 local information centers in the 26 governorates of Egypt that make plans for the family planning campaign. In 1992 the Minya Initiative, a family planning project was implemented in the Minya Governorate. As a result, the contraceptive prevalence rate rose from 22% to 30% over 18 months. A new project, the Select Village Project, was developed in 1993 that replicates the Minya Initiative on the village level in other governorates. This new project that was implemented in sixteen governorates.

  13. Meteorology of Jupiter's Equatorial Hot Spots and Plumes from Cassini

    NASA Technical Reports Server (NTRS)

    Choi, David Sanghun; Showman, Adam P.; Vasavada, Ashwin R.; Simon-Miller, Amy A.

    2013-01-01

    We present an updated analysis of Jupiter's equatorial meteorology from Cassini observations. For two months preceding the spacecraft's closest approach, the Imaging Science Subsystem (ISS) onboard regularly imaged the atmosphere. We created time-lapse movies from this period in order to analyze the dynamics of equatorial hot spots and their interactions with adjacent latitudes. Hot spots are relatively cloud-free regions that emit strongly at 5 lm; improved knowledge of these features is crucial for fully understanding Galileo probe measurements taken during its descent through one. Hot spots are quasistable, rectangular dark areas on visible-wavelength images, with defined eastern edges that sharply contrast with surrounding clouds, but diffuse western edges serving as nebulous boundaries with adjacent equatorial plumes. Hot spots exhibit significant variations in size and shape over timescales of days and weeks. Some of these changes correspond with passing vortex systems from adjacent latitudes interacting with hot spots. Strong anticyclonic gyres present to the south and southeast of the dark areas appear to circulate into hot spots. Impressive, bright white plumes occupy spaces in between hot spots. Compact cirrus-like 'scooter' clouds flow rapidly through the plumes before disappearing within the dark areas. These clouds travel at 150-200 m/s, much faster than the 100 m/s hot spot and plume drift speed. This raises the possibility that the scooter clouds may be more illustrative of the actual jet stream speed at these latitudes. Most previously published zonal wind profiles represent the drift speed of the hot spots at their latitude from pattern matching of the entire longitudinal image strip. If a downward branch of an equatorially-trapped Rossby wave controls the overall appearance of hot spots, however, the westward phase velocity of the wave leads to underestimates of the true jet stream speed.

  14. Maize Genetics and Genomics Database

    Technology Transfer Automated Retrieval System (TEKTRAN)

    This report is provided each year to our stakeholders in the maize genetic community. In this report, we describe the five-year plan for MaizeGDB reviewed in early 2008 by the USDA-ARS peer review process and which was developed with inputs from our Working Group and the Allerton 2007 Report (MNL 82...

  15. Maize variety and method of production

    DOEpatents

    Pauly, Markus; Hake, Sarah; Kraemer, Florian J

    2014-05-27

    The disclosure relates to a maize plant, seed, variety, and hybrid. More specifically, the disclosure relates to a maize plant containing a Cal-1 allele, whose expression results in increased cell wall-derived glucan content in the maize plant. The disclosure also relates to crossing inbreds, varieties, and hybrids containing the Cal-1 allele to produce novel types and varieties of maize plants.

  16. The MaizeGDB Genome Browser

    Technology Transfer Automated Retrieval System (TEKTRAN)

    MaizeGDB (http://www.maizegdb.org) is the community database for maize genetics and genomics. As part of an effort to develop MaizeGDB as a more sequence-centric resource, we implemented a genome browser based on information we gathered by surveying the community of maize geneticists. Based on commu...

  17. Metabolic pathway resources at MaizeGDB

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Two maize metabolic networks are available at MaizeGDB: MaizeCyc (http://maizecyc.maizegdb.org, also at Gramene) and CornCyc (http://corncyc.maizegdb.org, also at the Plant Metabolic Network). MaizeCyc was developed by Gramene, and CornCyc by the Plant Metabolic Network, both in collaboration with M...

  18. Thc6 protein, isolated from Trichoderma harzianum, can induce maize defense response against Curvularia lunata.

    PubMed

    Fan, Lili; Fu, Kehe; Yu, Chuanjin; Li, Yingying; Li, Yaqian; Chen, Jie

    2015-05-01

    Mutant T66 was isolated from 450 mutants (constructed with Agrobacterium tumefaciens-mediated transformation method) of Trichoderma harzianum. Maize seeds coated with T66 were more susceptible to Curvularia lunata when compared with those coated with wild-type (WT) strain. The disease index of maize treated with T66 and WT were 62.5 and 42.1%, respectively. Further research showed T-DNA has inserted into the ORF of one gene, which resulted in the functional difference between WT and T66. The gene was cloned and named Thc6, which encodes a novel 327 amino acid protein. To investigate its function, we obtained knockout, complementation, and overexpression mutants of Thc6. Challenge inoculation studies suggested that the Thc6 overexpression mutant can reduce the disease index of maize inbred line Huangzao 4 against the leaf spot pathogen (C. lunata). Meanwhile, The Thc6 mutants were found to affect the resistance of maize inbred line Huangzao 4 against C. lunata by enhancing the activation of jasmonate-responsive genes expression. Liquid chromatography-mass spectrometry (LC-MS) data further confirmed that the concentration of jasmonate in the induced maize exhibits a parallel change tendency with the expression level of defense-related genes. Hence, the Thc6 gene could be participated in the induced resistance of maize inbred line Huangzao 4 against C. lunata infection through a jasmonic acid-dependent pathway.

  19. Analysis of the Dynamics of Jupiter's SSTB Spots

    NASA Astrophysics Data System (ADS)

    Morales-Juberias, R.

    2006-12-01

    In the last decade, three of the larger anti-cyclonic spots in Jupiter's atmosphere, the White Ovals (WOS), merged into a single large oval rivaling in size (and recently in color) with the Great Red Spot (GRS). The observation and modeling of these events allowed us to gain some insight on the nature and dynamics of these features; yet, several important questions still remain unanswered. Is the evolution of the WOS representative of how the GRS came to be? To what extent, if any, is the dynamics of the spots in one latitudinal domain linked to the dynamics of spots in adjacent latitudinal domains? What is the role in the merger events of the cyclonic cells generally observed to be embedded between the anti-cyclones? Is the merger of the WOS a sign of a global weather cycle on Jupiter, like has been recently proposed? Similar mergers to those observed between the WOS have been reported to occur among the smaller white spots in the region immediately to the south, historically known as the South South Temperate Belt (SSTB). This region has one of the higher densities of spots per longitude in the planet, and a detailed study of its morphology and evolution should help us address the questions raised by the mergers of the WOS. We present an analysis of the morphology and dynamics of the vortices in this region based mostly on HST archived images, with support of some ground based images from the Association of Lunar and Planetary Observers of Japan. Numerical simulations in which we explore the observed long-term evolution of the spots in the framework of vortex-streets are also presented. Finally, we try to reproduce the details observed in merger events between spots in this region. This work was supported by New Mexico Institute of Mining and Technology new faculty start-up funds.

  20. Regulation of pathogenesis by light in Cercospora zeae-maydis: Identification of a photoreceptor required for infection of maize

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Cercospora zeae-maydis causes gray leaf spot, one of the most prevalent and damaging foliar diseases of maize throughout the world. Light plays a key role in the virulence of C. zeae-maydis by inducing the biosynthesis of cercosporin, a photosensitizing, non-host-specific phytotoxin. We determined t...

  1. Measuring microfocal spots using digital radiography

    SciTech Connect

    Fry, David A; Ewert, Uwe

    2009-01-01

    Measurement of microfocus spot size can be important for several reasons: (1) Quality assurance during manufacture of microfocus tubes; (2) Tracking performance and stability of microfocus tubes; (3) Determining magnification is especially important for digital radiography where the native spatial resolution of the digital system is not adequate for the application; and (4) Knowledge of unsharpness from the focal spot alone. The European Standard EN 12543-5 is based on a simple geometrical method of calculating focal spot size from unsharpness of high magnification film radiographs. The following equations are used for the focal spot size measurement: By similar triangles the following equations are presupposed: f/a = U/b and M = (a+b)/a. These equations can be combined to yield the well known expression: U = f(M - 1). Solving for f, f = U/(M-1). Therefore, the focal spot size, f, can be calculated by measuring the radiographic unsharpness and magnification of a known object. This is the basis for these tests. The European standard actually uses one-half of the unsharpness (which are then added together) from both sides of the object to avoid additional unsharpness contributions due to edge transmission unsharpness of the round test object (the outside of the object is measured). So the equation becomes f = (1/2 U{sub 1} + 1/2 U{sub 2})/(M-1). In practice 1/2 U is measured from the 50% to the 90% signal points on the transition profile from ''black'' to ''white,'' (positive image) or attenuated to unattenuated portion of the image. The 50% to 90% points are chosen as a best fit to an assumed Gaussian radiation distribution from the focal spot and to avoid edge transmission effects. 1/2 U{sub 1} + 1/2 U{sub 2} corresponds about to the full width at half height of a Gaussian focal spot. A highly absorbing material (Tungsten, Tungsten Alloy, or Platinum) is used for the object. Either wires or a sphere are used as the object to eliminate alignment issues. One

  2. The Little Red Spot: Closest View Yet

    NASA Technical Reports Server (NTRS)

    2007-01-01

    This is a mosaic of three New Horizons images of Jupiter's Little Red Spot, taken with the spacecraft's Long Range Reconnaissance Imager (LORRI) camera at 17:41 Universal Time on February 26 from a range of 3.5 million kilometers (2.1 million miles). The image scale is 17 kilometers (11 miles) per pixel, and the area covered measures 33,000 kilometers (20,000 miles) from top to bottom, two and one-half times the diameter of Earth.

    The Little Red Spot, a smaller cousin of the famous Great Red Spot, formed in the past decade from the merger of three smaller Jovian storms, and is now the second-largest storm on Jupiter. About a year ago its color, formerly white, changed to a reddish shade similar to the Great Red Spot, perhaps because it is now powerful enough to dredge up reddish material from deeper inside Jupiter. These are the most detailed images ever taken of the Little Red Spot since its formation, and will be combined with even sharper images taken by New Horizons 10 hours later to map circulation patterns around and within the storm.

    LORRI took the images as the Sun was about to set on the Little Red Spot. The LORRI camera was designed to look at Pluto, where sunlight is much fainter than it is at Jupiter, so the images would have been overexposed if LORRI had looked at the storm when it was illuminated by the noonday Sun. The dim evening illumination helped the LORRI camera obtain well-exposed images. The New Horizons team used predictions made by amateur astronomers in 2006, based on their observations of the motion of the Little Red Spot with backyard telescopes, to help them accurately point LORRI at the storm.

    These are among a handful of Jupiter system images already returned by New Horizons during its close approach to Jupiter. Most of the data being gathered by the spacecraft are stored onboard and will be downlinked to Earth during March and April 2007.

  3. Poisson Spot with Magnetic Levitation

    ERIC Educational Resources Information Center

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

    2010-01-01

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

  4. Advances in Maize Transformation Technologies and Development of Transgenic Maize

    PubMed Central

    Yadava, Pranjal; Abhishek, Alok; Singh, Reeva; Singh, Ishwar; Kaul, Tanushri; Pattanayak, Arunava; Agrawal, Pawan K.

    2017-01-01

    Maize is the principal grain crop of the world. It is also the crop where genetic engineering has been employed to a great extent to improve its various traits. The ability to transform maize is a crucial step for application of gene technology in maize improvement. There have been constant improvements in the maize transformation technologies over past several years. The choice of genotype and the explant material to initiate transformation and the different types of media to be used in various stages of tissue culture can have significant impact on the outcomes of the transformation efforts. Various methods of gene transfer, like the particle bombardment, protoplast transformation, Agrobacterium-mediated, in planta transformation, etc., have been tried and improved over years. Similarly, various selection systems for retrieval of the transformants have been attempted. The commercial success of maize transformation and transgenic development is unmatched by any other crop so far. Maize transformation with newer gene editing technologies is opening up a fresh dimension in transformation protocols and work-flows. This review captures the various past and recent facets in improvement in maize transformation technologies and attempts to present a comprehensive updated picture of the current state of the art in this area. PMID:28111576

  5. Rocky Mountain spotted fever.

    PubMed

    Lacz, N L; Schwartz, R A; Kapila, R

    2006-04-01

    Rocky Mountain spotted fever (RMSF) is an unusual but important dermatological condition to identify without hesitation. The classic triad of headache, fever, and a rash that begins on the extremities and travels proximally to involve the trunk is found in a majority of patients. The cutaneous centripetal pattern is a result of cell to cell migration by the causative organism Rickettsia rickettsii. Such individuals should receive prompt antimicrobial therapy and supportive care to avoid serious and potentially fatal complications.

  6. Rocky Mountain spotted fever.

    PubMed

    Kamper, C A; Chessman, K H; Phelps, S J

    1988-02-01

    The epidemiology, pathogenesis, clinical features, and treatment of Rocky Mountain spotted fever are reviewed. Rocky Mountain spotted fever is a severe infection caused by Rickettsia rickettsii transmitted to man by various species of ticks. High-incidence areas exist in the southeast and south central United States. Only 60-70% of patients with the disease report a history of tick bite or exposure to tick-infested areas. The disease is initially characterized by fever, headache, gastrointestinal complaints, myalgia, and a generalized rash. In several days generalized vasculitis may lead to periorbital edema and nonpitting edema of the face and extremities. Central nervous system involvement is common. Because signs and symptoms associated with the disease are nonspecific, the diagnosis is often delayed or missed. Traditionally diagnostic confirmation relied on serologic testing, but an indirect fluorescent antibody assay will soon be commercially available. Rocky Mountain spotted fever is usually treated with the rickettsiostatic agents chloramphenicol or tetracycline, but few comparative data on these agents in patients with the disease are available. For patients who cannot tolerate oral medications, intravenous chloramphenicol sodium succinate is the preferred treatment; chloramphenicol is also the drug of choice for children less than eight years of age. Otherwise, oral tetracycline hydrochloride is the drug of choice. Antibiotic therapy should be continued for 7-10 days or until the patient is afebrile for two to five days. All cases of Rocky Mountain spotted fever must be reported to the Centers for Disease Control. The best ways to decrease the morbidity and mortality of the disease are to increase awareness of its signs and symptoms and to prevent exposure to ticks.

  7. Predicting maize yield in Zimbabwe using dry dekads derived from remotely sensed Vegetation Condition Index

    NASA Astrophysics Data System (ADS)

    Kuri, Farai; Murwira, Amon; Murwira, Karin S.; Masocha, Mhosisi

    2014-12-01

    Maize is a key crop contributing to food security in Southern Africa yet accurate estimates of maize yield prior to harvesting are scarce. Timely and accurate estimates of maize production are essential for ensuring food security by enabling actionable mitigation strategies and policies for prevention of food shortages. In this study, we regressed the number of dry dekads derived from VCI against official ground-based maize yield estimates to generate simple linear regression models for predicting maize yield throughout Zimbabwe over four seasons (2009-10, 2010-11, 2011-12, and 2012-13). The VCI was computed using Normalized Difference Vegetation Index (NDVI) time series dataset from the SPOT VEGETATION sensor for the period 1998-2013. A significant negative linear relationship between number of dry dekads and maize yield was observed in each season. The variation in yield explained by the models ranged from 75% to 90%. The models were evaluated with official ground-based yield data that was not used to generate the models. There is a close match between the predicted yield and the official yield statistics with an error of 33%. The observed consistency in the negative relationship between number of dry dekads and ground-based estimates of maize yield as well as the high explanatory power of the regression models suggest that VCI-derived dry dekads could be used to predict maize yield before the end of the season thereby making it possible to plan strategies for dealing with food deficits or surpluses on time.

  8. MaizeGDB: The Maize Genetics and Genomics Database.

    PubMed

    Harper, Lisa; Gardiner, Jack; Andorf, Carson; Lawrence, Carolyn J

    2016-01-01

    MaizeGDB is the community database for biological information about the crop plant Zea mays. Genomic, genetic, sequence, gene product, functional characterization, literature reference, and person/organization contact information are among the datatypes stored at MaizeGDB. At the project's website ( http://www.maizegdb.org ) are custom interfaces enabling researchers to browse data and to seek out specific information matching explicit search criteria. In addition, pre-compiled reports are made available for particular types of data and bulletin boards are provided to facilitate communication and coordination among members of the community of maize geneticists.

  9. Provitamin A potential of landrace orange maize variety (Zea mays L.) grown in different geographical locations of central Malawi.

    PubMed

    Hwang, Taeyoung; Ndolo, Victoria U; Katundu, Mangani; Nyirenda, Blessings; Bezner-Kerr, Rachel; Arntfield, Susan; Beta, Trust

    2016-04-01

    The provitamin A potential of landrace orange maize from different locations (A, B, C and D) of central Malawi has been evaluated. Physicochemical compositions, color, total carotenoid content (TCC), carotenoid profiles, and oxygen radical absorbance capacity (ORAC) and 2,2-diphenyl-1-picryhydrazyl (DPPH) free radical scavenging activity as antioxidant capacities of maize were determined. Color values of orange maize had correlations with β-cryptoxanthin (r>0.36). TCC of white and orange maize averaged 2.12 and 59.5 mg/kg, respectively. Lutein was the most abundant carotenoid (47.8%) in orange maize, followed by zeaxanthin (24.2%), β-carotene (16.4%) and β-cryptoxanthin (11.6%). Location D showed the highest levels of lutein, zeaxanthin and antioxidant capacity. Provitamin A content of orange maize met the target level (15 μg/g) of biofortification. Retinol activity equivalent (RAE) from β-cryptoxanthin and β-carotene in orange maize averaged 81.73 μg/100g. In conclusion, orange maize has the potential to be a natural source of provitamin A.

  10. Ceres' Yellow Spots - Observations with Dawn Framing Camera

    NASA Astrophysics Data System (ADS)

    Schäfer, Michael; Schäfer, Tanja; Cloutis, Edward A.; Izawa, Matthew R. M.; Platz, Thomas; Castillo-Rogez, Julie C.; Hoffmann, Martin; Thangjam, Guneshwar S.; Kneissl, Thomas; Nathues, Andreas; Mengel, Kurt; Williams, David A.; Kallisch, Jan; Ripken, Joachim; Russell, Christopher T.

    2016-04-01

    The Framing Camera (FC) onboard the Dawn spacecraft acquired several spectral data sets of (1) Ceres with increasing spatial resolution (up to 135 m/pixel with nearly global coverage). The FC is equipped with seven color filters (0.4-1.0 μm) plus one panchromatic ('clear') filter [1]. We produced spectral mosaics using photometrically corrected FC color filter images as described in [2]. Even early FC color mosaics obtained during Dawn's approach unexpectedly exhibited quite a diversity of surface materials on Ceres. Besides the ordinary cerean surface material, potentially composed of ammoniated phyllosilicates [3] or some other alteration product of carbonaceous chondrites [4], a large number of bright spots were found on Ceres [5]. These spots are substantially brighter than the average surface (exceeding its triple standard deviation), with the spots within Occator crater being the brightest and most prominent examples (reflectance more than 10 times the average of Ceres). We observed bright spots which are different by their obvious yellow color. This yellow color appears both in a 'true color' RGB display (R=0.65, G=0.55, B=0.44 μm) as well as in a false color display (R=0.97, G=0.75, B=0.44 μm) using a linear 2% stretch. Their spectra show a steep red slope between 0.44 and 0.55 μm (UV drop-off). On the contrary to these yellow spots, the vast majority of bright spots appears white in the aforementioned color displays and exhibit blue sloped spectra, except for a shallow UV drop-off. Thus, yellow spots are easily distinguishable from white spots and the remaining cerean surface by their high values in the ratio 0.55/0.44 μm. We found 8 occurrences of yellow spots on Ceres. Most of them (>70 individual spots) occur both inside and outside crater Dantu, where white spots are also found in the immediate vicinity. Besides Dantu, further occurrences with only a few yellow spots were found at craters Ikapati and Gaue. Less definite occurrences are found at 97

  11. Turbulent Region Near Jupiter's Great Red Spot

    NASA Technical Reports Server (NTRS)

    1997-01-01

    True and false color mosaics of the turbulent region west of Jupiter's Great Red Spot. The Great Red Spot is on the planetary limb on the right hand side of each mosaic. The region west (left) of the Great Red Spot is characterized by large, turbulent structures that rapidly change in appearance. The turbulence results from the collision of a westward jet that is deflected northward by the Great Red Spot into a higher latitude eastward jet. The large eddies nearest to the Great Red Spot are bright, suggesting that convection and cloud formation are active there.

    The top mosaic combines the violet (410 nanometers) and near infrared continuum (756 nanometers) filter images to create a mosaic similar to how Jupiter would appear to human eyes. Differences in coloration are due to the composition and abundance of trace chemicals in Jupiter's atmosphere. The lower mosaic uses the Galileo imaging camera's three near-infrared (invisible) wavelengths (756 nanometers, 727 nanometers, and 889 nanometers displayed in red, green, and blue) to show variations in cloud height and thickness. Light blue clouds are high and thin, reddish clouds are deep, and white clouds are high and thick. Purple most likely represents a high haze overlying a clear deep atmosphere. Galileo is the first spacecraft to distinguish cloud layers on Jupiter.

    The mosaic is centered at 16.5 degrees south planetocentric latitude and 85 degrees west longitude. The north-south dimension of the Great Red Spot is approximately 11,000 kilometers. The smallest resolved features are tens of kilometers in size. North is at the top of the picture. The images used were taken on June 26, 1997 at a range of 1.2 million kilometers (1.05 million miles) by the Solid State Imaging (SSI) system on NASA's Galileo spacecraft.

    The Jet Propulsion Laboratory, Pasadena, CA manages the Galileo mission for NASA's Office of Space Science, Washington, DC. JPL is an operating division of California Institute of Technology

  12. [Rocky Mountain spotted fever].

    PubMed

    Reinauer, K M; Jaschonek, K; Kusch, G; Heizmann, W R; Döller, P C; Jenss, H

    1990-01-12

    After returning from a holiday in the USA a 24-year-old man fell ill with diarrhoea, high fever and marked rash including the palms of the hands and soles of the feet. When a history of a tick bite in the USA was elicited, a rickettsial infection was suspected. Treatment with doxycycline, 100 mg twice daily, was instituted finally and the fever slowly resolved. The patient became completely well again within four weeks. Serological tests confirmed the diagnosis of Rocky Mountain spotted fever.

  13. Dark Spots and Fans

    NASA Technical Reports Server (NTRS)

    2006-01-01

    As winter turns to spring at the south polar ice cap of Mars, the rising sun reveals dark spots and fans emerging from the cold polar night. Using visual images (left) and temperature data (right) from the Thermal Emission Imaging system on NASA's Mars Odyssey orbiter, scientists have built a new model for the origin of the dark markings. Scientists propose the markings come from dark sand and dust strewn by high-speed jets of carbon-dioxide gas. These erupt from under a layer of carbon-dioxide ice that forms each Martian winter.

  14. Spots and White Light Flares in an L Dwarf

    DTIC Science & Technology

    2013-01-01

    Program GN-2012A-Q-37) GMOS spectrograph (Hook et al. 2004) when a series of flares occurred. A spectrum of the most powerful flare in its impulsive...10:14 Hα HeI HeI HeI OI Fig. 4. Gemini-North GMOS spectra of W1906+40 in quiescence (below) and in flare. Note the broad Hα, atomic emission lines

  15. Configurable hot spot fixing system

    NASA Astrophysics Data System (ADS)

    Kajiwara, Masanari; Kobayashi, Sachiko; Mashita, Hiromitsu; Aburada, Ryota; Furuta, Nozomu; Kotani, Toshiya

    2014-03-01

    Hot spot fixing (HSF) method has been used to fix many hot spots automatically. However, conventional HSF based on a biasing based modification is difficult to fix many hot spots under a low-k1 lithography condition. In this paper we proposed a new HSF, called configurable hotspot fixing system. The HSF has two major concepts. One is a new function to utilize vacant space around a hot spot by adding new patterns or extending line end edges around the hot spot. The other is to evaluate many candidates at a time generated by the new functions. We confirmed the proposed HSF improves 73% on the number of fixing hot spots and reduces total fixing time by 50% on a device layout equivalent to 28nm-node. The result shows the proposed HSF is effective for layouts under the low-k1 lithography condition.

  16. Turbulent spots in hypervelocity flow

    NASA Astrophysics Data System (ADS)

    Jewell, Joseph S.; Leyva, Ivett A.; Shepherd, Joseph E.

    2017-04-01

    The turbulent spot propagation process in boundary layer flows of air, nitrogen, carbon dioxide, and air/carbon dioxide mixtures in thermochemical nonequilibrium at high enthalpy is investigated. Experiments are performed in a hypervelocity reflected shock tunnel with a 5-degree half-angle axisymmetric cone instrumented with flush-mounted fast-response coaxial thermocouples. Time-resolved and spatially demarcated heat transfer traces are used to track the propagation of turbulent bursts within the mean flow, and convection rates at approximately 91, 74, and 63% of the boundary layer edge velocity, respectively, are observed for the leading edge, peak, and trailing edge of the spots. A simple model constructed with these spot propagation parameters is used to infer spot generation rates from observed transition onset to completion distance. Spot generation rates in air and nitrogen are estimated to be approximately twice the spot generation rates in air/carbon dioxide mixtures.

  17. Sequencing the maize genome.

    PubMed

    Martienssen, Robert A; Rabinowicz, Pablo D; O'Shaughnessy, Andrew; McCombie, W Richard

    2004-04-01

    Sequencing of complex genomes can be accomplished by enriching shotgun libraries for genes. In maize, gene-enrichment by copy-number normalization (high C(0)t) and methylation filtration (MF) have been used to generate up to two-fold coverage of the gene-space with less than 1 million sequencing reads. Simulations using sequenced bacterial artificial chromosome (BAC) clones predict that 5x coverage of gene-rich regions, accompanied by less than 1x coverage of subclones from BAC contigs, will generate high-quality mapped sequence that meets the needs of geneticists while accommodating unusually high levels of structural polymorphism. By sequencing several inbred strains, we propose a strategy for capturing this polymorphism to investigate hybrid vigor or heterosis.

  18. Biofortified orange maize enhances β-cryptoxanthin concentrations in egg yolks of laying hens better than tangerine peel fortificant.

    PubMed

    Heying, Emily K; Tanumihardjo, Jacob P; Vasic, Vedran; Cook, Mark; Palacios-Rojas, Natalia; Tanumihardjo, Sherry A

    2014-12-10

    The xanthophyll β-cryptoxanthin provides vitamin A and has other purported health benefits. Laying hens deposit xanthophyll carotenoids into egg yolk. Hens (n = 8/group) were fed conventional-bred high β-cryptoxanthin biofortified (orange) maize, tangerine peel-fortified white maize, lutein-fortified yellow maize, or white maize for 40 d to investigate yolk color changes using L*a*b* scales, yolk carotenoid enhancement, and hen vitamin A status. Yolks from hens fed orange maize had scores indicating a darker, orange color and mean higher β-cryptoxanthin, zeaxanthin, and β-carotene concentrations (8.43 ± 1.82, 23.1 ± 4.8, 0.16 ± 0.08 nmol/g, respectively) than other treatments (P < 0.0001). Yolk retinol concentrations (mean: 14.4 ± 3.42 nmol/g) were similar among groups and decreased with time (P < 0.0001). Hens fed orange maize had higher liver retinol (0.53 ± 0.20 μmol/g liver) than other groups (P < 0.0001). β-Cryptoxanthin-biofortified eggs could be another choice for consumers, providing enhanced color through a provitamin A carotenoid and supporting eggs' status as a functional food.

  19. Comparative proteomic analyses provide new insights into low phosphorus stress responses in maize leaves.

    PubMed

    Zhang, Kewei; Liu, Hanhan; Tao, Peilin; Chen, Huan

    2014-01-01

    Phosphorus deficiency limits plant growth and development. To better understand the mechanisms behind how maize responds to phosphate stress, we compared the proteome analysis results of two groups of maize leaves that were treated separately with 1,000 µM (control, +P) and 5 µM of KH2PO4 (intervention group, -P) for 25 days. In total, 1,342 protein spots were detected on 2-DE maps and 15.43% had changed (P<0.05; ≥1.5-fold) significantly in quantity between the +P and -P groups. These proteins are involved in several major metabolic pathways, including photosynthesis, carbohydrate metabolism, energy metabolism, secondary metabolism, signal transduction, protein synthesis, cell rescue and cell defense and virulence. The results showed that the reduction in photosynthesis under low phosphorus treatment was due to the down-regulation of the proteins involved in CO2 enrichment, the Calvin cycle and the electron transport system. Electron transport and photosynthesis restrictions resulted in a large accumulation of peroxides. Maize has developed many different reactive oxygen species (ROS) scavenging mechanisms to cope with low phosphorus stress, including up-regulating its antioxidant content and antioxidase activity. After being subjected to phosphorus stress over a long period, maize may increase its internal phosphorus utilization efficiency by altering photorespiration, starch synthesis and lipid composition. These results provide important information about how maize responds to low phosphorus stress.

  20. Comparative Proteomic Analyses Provide New Insights into Low Phosphorus Stress Responses in Maize Leaves

    PubMed Central

    Zhang, Kewei; Liu, Hanhan; Tao, Peilin; Chen, Huan

    2014-01-01

    Phosphorus deficiency limits plant growth and development. To better understand the mechanisms behind how maize responds to phosphate stress, we compared the proteome analysis results of two groups of maize leaves that were treated separately with 1,000 µM (control, +P) and 5 µM of KH2PO4 (intervention group, −P) for 25 days. In total, 1,342 protein spots were detected on 2-DE maps and 15.43% had changed (P<0.05; ≥1.5-fold) significantly in quantity between the +P and −P groups. These proteins are involved in several major metabolic pathways, including photosynthesis, carbohydrate metabolism, energy metabolism, secondary metabolism, signal transduction, protein synthesis, cell rescue and cell defense and virulence. The results showed that the reduction in photosynthesis under low phosphorus treatment was due to the down-regulation of the proteins involved in CO2 enrichment, the Calvin cycle and the electron transport system. Electron transport and photosynthesis restrictions resulted in a large accumulation of peroxides. Maize has developed many different reactive oxygen species (ROS) scavenging mechanisms to cope with low phosphorus stress, including up-regulating its antioxidant content and antioxidase activity. After being subjected to phosphorus stress over a long period, maize may increase its internal phosphorus utilization efficiency by altering photorespiration, starch synthesis and lipid composition. These results provide important information about how maize responds to low phosphorus stress. PMID:24858307

  1. The Evolution of Modulated Wavetrains Into Turbulent Spots

    NASA Technical Reports Server (NTRS)

    Gaster, M.

    2007-01-01

    Experiment are being carried out to study the process by which th almost periodic disturbance waves generated naturally by the freestream evolve into turbulence. The boundary layer on a flat plate has been used for this study. The novelty of the approach is in the form of artificial excitation that is used. In this work the flow is excited artificially by deterministic white noise. The weak T-S wave created develops down stream, becomes nonlinear and blows up locally onto a highly distorted flow. These large local distortions of the mean flow allow very high frequency disturbances to grow and form into small turbulent spots. The spots arise from the excitation, and if the same noise sequence is repeated a spot will form at the same position and time instant relative to the excitation.

  2. Influence of maize/lablab intercropping on lepidopterous stem borer infestation in maize.

    PubMed

    Maluleke, Mary H; Addo-Bediako, Abraham; Ayisi, Kingsley K

    2005-04-01

    Lepidopterous stem borers seriously affect production of maize, Zea mays L., in sub-Saharan Africa. Intercropping maize with legumes such as lablab, Lablab purpurens (L.), is one of the effective systems to control stem borers. Sole culture maize and maize/lablab intercrop system of different lablab densities were planted at two locations to investigate the effects of intercrop system on incidence and severity of stem borers with particular reference to Chilo partellus (Swinhoe) (Lepidoptera: Pyralidae). Stem borer infestation was found to be more severe in sole culture maize than maize in maize/lablab intercrop. There was a significantly negative relationship between lablab densities and maize grain yields, suggesting a possible competition for resources between the two crops. It was concluded that density of lablab and date of planting of lablab in maize/lablab intercropping have significant affects on stem borer populations and maize grain yields.

  3. White Rock

    NASA Technical Reports Server (NTRS)

    2002-01-01

    (Released 19 April 2002) The Science 'White Rock' is the unofficial name for this unusual landform which was first observed during the Mariner 9 mission in the early 1970's. As later analysis of additional data sets would show, White Rock is neither white nor dense rock. Its apparent brightness arises from the fact that the material surrounding it is so dark. Images from the Mars Global Surveyor MOC camera revealed dark sand dunes surrounding White Rock and on the floor of the troughs within it. Some of these dunes are just apparent in the THEMIS image. Although there was speculation that the material composing White Rock could be salts from an ancient dry lakebed, spectral data from the MGS TES instrument did not support this claim. Instead, the White Rock deposit may be the erosional remnant of a previously more continuous occurrence of air fall sediments, either volcanic ash or windblown dust. The THEMIS image offers new evidence for the idea that the original deposit covered a larger area. Approximately 10 kilometers to the southeast of the main deposit are some tiny knobs of similarly bright material preserved on the floor of a small crater. Given that the eolian erosion of the main White Rock deposit has produced isolated knobs at its edges, it is reasonable to suspect that the more distant outliers are the remnants of a once continuous deposit that stretched at least to this location. The fact that so little remains of the larger deposit suggests that the material is very easily eroded and simply blows away. The Story Fingers of hard, white rock seem to jut out like icy daggers across a moody Martian surface, but appearances can be deceiving. These bright, jagged features are neither white, nor icy, nor even hard and rocky! So what are they, and why are they so different from the surrounding terrain? Scientists know that you can't always trust what your eyes see alone. You have to use other kinds of science instruments to measure things that our eyes can

  4. Resolving stellar surface spots

    NASA Astrophysics Data System (ADS)

    Strassmeier, K. G.; Carroll, T.; Rice, J. B.; Savanov, I. S.

    Doppler imaging of stellar surfaces is a novel technique with similarities to medical brain tomography (instead of a fixed brain and a rotating scanner, astronomers have a fixed spectrograph and a rotating brain, star of course). The number of free (internal) parameters is of the order of the number of surface grid points and only constrained by the number of input data points. This obviously ill-posed situation requires modern inversion algorithms with penalty functions of the form of maximum entropy or Tikhonov etc.. We present a brief status review of our Doppler imaging codes at AIP that span from temperature and spot-filling-factor mapping to full Stokes-based magnetic field mapping.

  5. Rocky Mountain spotted fever.

    PubMed

    Dantas-Torres, Filipe

    2007-11-01

    Rocky Mountain spotted fever (RMSF) is a life-threatening disease caused by Rickettsia rickettsii, an obligately intracellular bacterium that is spread to human beings by ticks. More than a century after its first clinical description, this disease is still among the most virulent human infections identified, being potentially fatal even in previously healthy young people. The diagnosis of RMSF is based on the patient's history and a physical examination, and often presents a dilemma for clinicians because of the non-specific presentation of the disease in its early course. Early empirical treatment is essential to prevent severe complications or a fatal outcome, and treatment should be initiated even in unconfirmed cases. Because there is no vaccine available against RMSF, avoidance of tick-infested areas is still the best way to prevent the infection.

  6. Inbreeding drives maize centromere evolution.

    PubMed

    Schneider, Kevin L; Xie, Zidian; Wolfgruber, Thomas K; Presting, Gernot G

    2016-02-23

    Functional centromeres, the chromosomal sites of spindle attachment during cell division, are marked epigenetically by the centromere-specific histone H3 variant cenH3 and typically contain long stretches of centromere-specific tandem DNA repeats (∼1.8 Mb in maize). In 23 inbreds of domesticated maize chosen to represent the genetic diversity of maize germplasm, partial or nearly complete loss of the tandem DNA repeat CentC precedes 57 independent cenH3 relocation events that result in neocentromere formation. Chromosomal regions with newly acquired cenH3 are colonized by the centromere-specific retrotransposon CR2 at a rate that would result in centromere-sized CR2 clusters in 20,000-95,000 y. Three lines of evidence indicate that CentC loss is linked to inbreeding, including (i) CEN10 of temperate lineages, presumed to have experienced a genetic bottleneck, contain less CentC than their tropical relatives; (ii) strong selection for centromere-linked genes in domesticated maize reduced diversity at seven of the ten maize centromeres to only one or two postdomestication haplotypes; and (iii) the centromere with the largest number of haplotypes in domesticated maize (CEN7) has the highest CentC levels in nearly all domesticated lines. Rare recombinations introduced one (CEN2) or more (CEN5) alternate CEN haplotypes while retaining a single haplotype at domestication loci linked to these centromeres. Taken together, this evidence strongly suggests that inbreeding, favored by postdomestication selection for centromere-linked genes affecting key domestication or agricultural traits, drives replacement of the tandem centromere repeats in maize and other crop plants. Similar forces may act during speciation in natural systems.

  7. MaizeGDB: New tools and resource

    Technology Transfer Automated Retrieval System (TEKTRAN)

    MaizeGDB, the USDA-ARS genetics and genomics database, is a highly curated, community-oriented informatics service to researchers focused on the crop plant and model organism Zea mays. MaizeGDB facilitates maize research by curating, integrating, and maintaining a database that serves as the central...

  8. The Genetic Architecture of Maize Flowering Time

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Flowering time is the key trait controlling adaptation of plants to their local environment, and, in an outcrossing species like maize, it is a complex trait. Variation for this complex trait was dissected in maize using a novel set of 5000 recombinant inbred lines (maize Nested Association Mapping...

  9. MaizeGDB's New Genome Browser Project

    Technology Transfer Automated Retrieval System (TEKTRAN)

    MaizeGDB (http://www.maizegdb.org) is the community database for maize genetics and genomics. Based upon the 2006 MaizeGDB Working Group Report (available at http://www.maizegdb.org/working_group.php) and the Allerton Report (http://www.maizegdb.org/AllertonReport.doc), it has become evident that th...

  10. Rocky Mountain spotted fever, Colombia.

    PubMed

    Hidalgo, Marylin; Orejuela, Leonora; Fuya, Patricia; Carrillo, Pilar; Hernandez, Jorge; Parra, Edgar; Keng, Colette; Small, Melissa; Olano, Juan P; Bouyer, Donald; Castaneda, Elizabeth; Walker, David; Valbuena, Gustavo

    2007-07-01

    We investigated 2 fatal cases of Rocky Mountain spotted fever that occurred in 2003 and 2004 near the same locality in Colombia where the disease was first reported in the 1930s. A retrospective serosurvey of febrile patients showed that > 21% of the serum samples had antibodies aaainst spotted fever group rickettsiae.

  11. The hot-spot environment of SW Sex in a low state

    NASA Astrophysics Data System (ADS)

    Groot, Paul J.; Rutten, René G. M.; van Paradijs, Jan

    2000-04-01

    Based on observations obtained with the 2.5m Isaac Newton Telescope we show that the characteristics of the SW Sex stars can be explained by the dominance of a `hot-spot' like feature in the accretion disk. In SW Sex this `hot-spot' region is located at a distance of 0.5 RL 1 from the white-dwarf and is best visible at phase ϕ=0.95. The location of the hot-spot as deduced from spectral eclipse mapping coincides with the formation site of the main emission lines. We deduce that this hot-spot region is formed by a shock, which we speculate to be the consequence of a high mass-transfer rate and a long spin-period of the non-magnetic white dwarf.

  12. White phosphorus

    Integrated Risk Information System (IRIS)

    White phosphorus ; CASRN 7723 - 14 - 0 Human health assessment information on a chemical substance is included in the IRIS database only after a comprehensive review of toxicity data , as outlined in the IRIS assessment development process . Sections I ( Health Hazard Assessments for Noncarcinogenic

  13. White Flight

    ERIC Educational Resources Information Center

    Russell, Christine

    1975-01-01

    Focuses on the issue of whether the implementation of school desegregation significantly increases the decline in percentage of resident white. Presents data from a study conducted in 86 Northern school districts from a 91-city study. Data were obtained from statistics published by the Department of Health, Education and Welfare beginning in 1967.…

  14. Hot Spot Cosmic Accelerators

    NASA Astrophysics Data System (ADS)

    2002-11-01

    length of more than 3 million light-years, or no less than one-and-a-half times the distance from the Milky Way to the Andromeda galaxy, this structure is indeed gigantic. The region where the jets collide with the intergalactic medium are known as " hot spots ". Superposing the intensity contours of the radio emission from the southern "hot spot" on a near-infrared J-band (wavelength 1.25 µm) VLT ISAAC image ("b") shows three distinct emitting areas; they are even better visible on the I-band (0.9 µm) FORS1 image ("c"). This emission is obviously associated with the shock front visible on the radio image. This is one of the first times it has been possible to obtain an optical/near-IR image of synchrotron emission from such an intergalactic shock and, thanks to the sensitivity and image sharpness of the VLT, the most detailed view of its kind so far . The central area (with the strongest emission) is where the plasma jet from the galaxy centre hits the intergalactic medium. The light from the two other "knots", some 10 - 15,000 light-years away from the central "hot spot", is also interpreted as synchrotron emission. However, in view of the large distance, the astronomers are convinced that it must be caused by electrons accelerated in secondary processes at those sites . The new images thus confirm that electrons are being continuously accelerated in these "knots" - hence called "cosmic accelerators" - far from the galaxy and the main jets, and in nearly empty space. The exact physical circumstances of this effect are not well known and will be the subject of further investigations. The present VLT-images of the "hot spots" near 3C 445 may not have the same public appeal as some of those beautiful images that have been produced by the same instruments during the past years. But they are not less valuable - their unusual importance is of a different kind, as they now herald the advent of fundamentally new insights into the mysteries of this class of remote and active

  15. Maize, tropical (Zea mays L.).

    PubMed

    Assem, Shireen K

    2015-01-01

    Maize (Zea mays L.) is the third most important food crop globally after wheat and rice. In sub-Saharan Africa, tropical maize has traditionally been the main staple of the diet; 95 % of the maize grown is consumed directly as human food and as an important source of income for the resource-poor rural population. The biotechnological approach to engineer biotic and abiotic traits implies the availability of an efficient plant transformation method. The production of genetically transformed plants depends both on the ability to integrate foreign genes into target cells and the efficiency with which plants are regenerated. Maize transformation and regeneration through immature embryo culture is the most efficient system to regenerate normal transgenic plants. However, this system is highly genotype dependent. Genotypes adapted to tropic areas are difficult to regenerate. Therefore, transformation methods used with model genotypes adapted to temperate areas are not necessarily efficient with tropical lines. Agrobacterium-mediated transformation is the method of choice since it has been first achieved in 1996. In this report, we describe a transformation method used successfully with several tropical maize lines. All the steps of transformation and regeneration are described in details. This protocol can be used with a wide variety of tropical lines. However, some modifications may be needed with recalcitrant lines.

  16. Fast and controlled release of triamcinolone acetonide from extrusion-spheronization pellets based on mixtures of native starch with dextrin or waxy maize starch.

    PubMed

    Sergio, Almeida-Prieto; Isabel, de Sá Ferreira da Rocha Cristiana; José, Blanco-Méndez; Otero-Espinar, Francisco J

    2007-09-01

    Pellets composed chiefly of inexpensive starches allow modulation of the rate of release of the poorly soluble drug triamcinolone acetonide in media of pH 1.2-6.8. Wheat- or maize-starch-based pellets with 20% of white dextrin release the drug in vitro almost completely within 20 min, while maize-starch-based pellets with 5-35% of waxy maize starch sustain gradual release over periods of 9-12 hr or longer when prepared using appropriate amounts of granulation fluid.

  17. Saturn's Hot Spot

    NASA Technical Reports Server (NTRS)

    2005-01-01

    This is the sharpest image of Saturn's temperature emissions taken from the ground; it is a mosaic of 35 individual exposures made at the W.M. Keck I Observatory, Mauna Kea, Hawaii on Feb. 4, 2004.

    The images to create this mosaic were taken with infrared radiation. The mosaic was taken at a wavelength near 17.65 microns and is sensitive to temperatures in Saturn's upper troposphere. The prominent hot spot at the bottom of the image is right at Saturn's south pole. The warming of the southern hemisphere was expected, as Saturn was just past southern summer solstice, but the abrupt changes in temperature with latitude were not expected. The tropospheric temperature increases toward the pole abruptly near 70 degrees latitude from 88 to 89 Kelvin (-301 to -299 degrees Fahrenheit) and then to 91 Kelvin (-296 degrees Fahrenheit) right at the pole.

    Ring particles are not at a uniform temperature everywhere in their orbit around Saturn. The ring particles are orbiting clockwise in this image. Particles are coldest just after having cooled down in Saturn's shadow (lower left). As they orbit Saturn, the particles increase in temperature up to a maximum (lower right) just before passing behind Saturn again in shadow.

    A small section of the ring image is missing because of incomplete mosaic coverage during the observing sequence.

  18. Global maize production, utilization, and consumption.

    PubMed

    Ranum, Peter; Peña-Rosas, Juan Pablo; Garcia-Casal, Maria Nieves

    2014-04-01

    Maize (Zea mays), also called corn, is believed to have originated in central Mexico 7000 years ago from a wild grass, and Native Americans transformed maize into a better source of food. Maize contains approximately 72% starch, 10% protein, and 4% fat, supplying an energy density of 365 Kcal/100 g and is grown throughout the world, with the United States, China, and Brazil being the top three maize-producing countries in the world, producing approximately 563 of the 717 million metric tons/year. Maize can be processed into a variety of food and industrial products, including starch, sweeteners, oil, beverages, glue, industrial alcohol, and fuel ethanol. In the last 10 years, the use of maize for fuel production significantly increased, accounting for approximately 40% of the maize production in the United States. As the ethanol industry absorbs a larger share of the maize crop, higher prices for maize will intensify demand competition and could affect maize prices for animal and human consumption. Low production costs, along with the high consumption of maize flour and cornmeal, especially where micronutrient deficiencies are common public health problems, make this food staple an ideal food vehicle for fortification.

  19. Single spots, unipolar magnetic regions, and pairs of spots

    NASA Astrophysics Data System (ADS)

    Akasofu, S.-I.

    2014-06-01

    McIntosh (1981) noted that sunspot pairs appear preferentially near the boundary of unipolar magnetic (UM) regions of opposite polarity. A large number of solar magnetograms from the Mount Wilson Observatory and the Kitt Peak Observatory during fairly quiet periods are examined to confirm his finding. In this study, it is also found collaterally that positive single spots appear in a positive UM region and vice versa. It is suggested thus that a pair of spots of opposite polarity is formed because two single spots develop in the vicinity of the boundary (the neutral line) of two UM regions of opposite polarity for polarity arrangement appropriate to the Hale law, namely, the Hale boundary. For these reasons, it is suggested that single spots and UM regions have significant meaning in solar magnetism.

  20. A chromosome inversion near the KIT gene and the Tobiano spotting pattern in horses.

    PubMed

    Brooks, S A; Lear, T L; Adelson, D L; Bailey, E

    2007-01-01

    Tobiano is a white spotting pattern in horses caused by a dominant gene, Tobiano(TO). Here, we report TO associated with a large paracentric chromosome inversion on horse chromosome 3. DNA sequences flanking the inversion were identified and a PCR test was developed to detect the inversion. The inversion was only found in horses with the tobiano pattern, including horses with diverse genetic backgrounds, which indicated a common genetic origin thousands of years ago. The inversion does not interrupt any annotated genes, but begins approximately 100 kb downstream of the KIT gene. This inversion may disrupt regulatory sequences for the KIT gene and cause the white spotting pattern.

  1. Maize-targeted mutagenesis: A knockout resource for maize.

    PubMed

    May, Bruce P; Liu, Hong; Vollbrecht, Erik; Senior, Lynn; Rabinowicz, Pablo D; Roh, Donna; Pan, Xiaokang; Stein, Lincoln; Freeling, Mike; Alexander, Danny; Martienssen, Rob

    2003-09-30

    We describe an efficient system for site-selected transposon mutagenesis in maize. A total of 43,776 F1 plants were generated by using Robertson's Mutator (Mu) pollen parents and self-pollinated to establish a library of transposon-mutagenized seed. The frequency of new seed mutants was between 10-4 and 10-5 per F1 plant. As a service to the maize community, maize-targeted mutagenesis selects insertions in genes of interest from this library by using the PCR. Pedigree, knockout, sequence, phenotype, and other information is stored in a powerful interactive database (maize-targeted mutagenesis database) that enables analysis of the entire population and the handling of knockout requests. By inhibiting Mu activity in most F1 plants, we sought to reduce somatic insertions that may cause false positives selected from pooled tissue. By monitoring the remaining Mu activity in the F2, however, we demonstrate that seed phenotypes depend on it, and false positives occur in lines that appear to lack it. We conclude that more than half of all mutations arising in this population are suppressed on losing Mu activity. These results have implications for epigenetic models of inbreeding and for functional genomics.

  2. Alightment of Spotted Wing Drosophila (Diptera: Drosophilidae) on Odorless Disks Varying in Color.

    PubMed

    Kirkpatrick, D M; McGhee, P S; Hermann, S L; Gut, L J; Miller, J R

    2016-02-01

    Methods for trapping spotted wing drosophila, Drosophila suzukii (Matsmura) (Diptera: Drosophilidae), have not yet been optimized for detecting this devastating pest of soft-skinned fruits. Here, we report outcomes of choice and no-choice laboratory bioassays quantifying the rates of spotted wing drosophila alightment on 5-cm-diameter sticky disks of various colors, but no fruit odors. Red, purple, and black disks captured the most spotted wing drosophila when presented against a white background. Male and female spotted wing drosophila responded identically in these tests. Significantly more D. suzukii were captured on the red and yellow disks than those presenting the corresponding grayscale for that color, proving that D. suzukii perceives colors and not just the level of target brightness. Fluorescent red is the best candidate for trap color, while clear and white are the least desirable. However, when the background was switched to black, all nonfluorescent colors were equally acceptable to spotted wing drosophila, suggesting that background must be specified when reporting spotted wing drosophila color preference. Additional spotted wing drosophila research is justified on the effects of target color against natural backgrounds.

  3. False Color Mosaic Great Red Spot

    NASA Technical Reports Server (NTRS)

    1996-01-01

    False color representation of Jupiter's Great Red Spot (GRS) taken through three different near-infrared filters of the Galileo imaging system and processed to reveal cloud top height. Images taken through Galileo's near-infrared filters record sunlight beyond the visible range that penetrates to different depths in Jupiter's atmosphere before being reflected by clouds. The Great Red Spot appears pink and the surrounding region blue because of the particular color coding used in this representation. Light reflected by Jupiter at a wavelength (886 nm) where methane strongly absorbs is shown in red. Due to this absorption, only high clouds can reflect sunlight in this wavelength. Reflected light at a wavelength (732 nm) where methane absorbs less strongly is shown in green. Lower clouds can reflect sunlight in this wavelength. Reflected light at a wavelength (757 nm) where there are essentially no absorbers in the Jovian atmosphere is shown in blue: This light is reflected from the deepest clouds. Thus, the color of a cloud in this image indicates its height. Blue or black areas are deep clouds; pink areas are high, thin hazes; white areas are high, thick clouds. This image shows the Great Red Spot to be relatively high, as are some smaller clouds to the northeast and northwest that are surprisingly like towering thunderstorms found on Earth. The deepest clouds are in the collar surrounding the Great Red Spot, and also just to the northwest of the high (bright) cloud in the northwest corner of the image. Preliminary modeling shows these cloud heights vary over 30 km in altitude. This mosaic, of eighteen images (6 in each filter) taken over a 6 minute interval during the second GRS observing sequence on June 26, 1996, has been map-projected to a uniform grid of latitude and longitude. North is at the top.

    Launched in October 1989, Galileo entered orbit around Jupiter on December 7, 1995. The spacecraft's mission is to conduct detailed studies of the giant planet

  4. Weird Warm Spot on Exoplanet

    NASA Video Gallery

    This animation illustrates an unexpected warm spot on the surface of a gaseous exoplanet. NASA's Spitzer Space Telescope discovered that the hottest part of the planet, shown here as bright, orange...

  5. Center Spot: Shoe Box Science

    ERIC Educational Resources Information Center

    Hoffman, Jan

    1976-01-01

    This is the second "Center Spot" devoted to Jan Hoffman's "Shoe Box Science," a program that organizes manipulative materials so that children can identify, describe, order, construct, name and distinguish on their own.

  6. Seedling development in maize cv. B73 and blue light-mediated proteomic changes in the tip vs. stem of the coleoptile.

    PubMed

    Deng, Zhiping; Wang, Zhi-Yong; Kutschera, Ulrich

    2016-09-15

    In 2009, the draft genome of the reference inbred line of maize (Zea mays L. spp. mays cv. B73) was published so that, using this specific corn variety, molecular analyses of physiological processes became possible. However, the morphology and developmental patterns of B73 maize, compared with that of the more frequently used hybrid varieties, have not yet been analyzed. Here, we describe organ development in seedlings of B73 maize and in those of six other hybrid cultivars, and document significant morphological as well as quantitative differences between these varieties of Z. mays. In a second set of experiments, we used etiolated seedlings of B73 maize to analyze the effect of blue light (BL) on the patterns of proteins in the tip vs. growing region of this sheath-like organ. By using two-dimensional difference gel electrophoresis (2D DIGE), coupled with tandem mass spectrometry, we detected, in the microsomal fraction of maize coleoptile tips, rapid changes in the abundance of protein spots of maize phototropin 1 and several metabolic enzymes. In the sub-apical (growing) region of the coleoptile, proteomic changes were less pronounced. These results suggest that the tip of the coleoptile of B73 maize may serve as a unique model system for dissecting BL responses in a light-sensitive plant organ of known function.

  7. Betaine Deficiency in Maize 1

    PubMed Central

    Lerma, Claudia; Rich, Patrick J.; Ju, Grace C.; Yang, Wen-Ju; Hanson, Andrew D.; Rhodes, David

    1991-01-01

    Maize (Zea mays L.) is a betaine-accumulating species, but certain maize genotypes lack betaine almost completely; a single recessive gene has been implicated as the cause of this deficiency (D Rhodes, PJ Rich [1988] Plant Physiol 88: 102-108). This study was undertaken to determine whether betaine deficiency in diverse maize germplasm is conditioned by the same genetic locus, and to define the biochemical lesion(s) involved. Complementation tests indicated that all 13 deficient genotypes tested shared a common locus. One maize population (P77) was found to be segregating for betaine deficiency, and true breeding individuals were used to produce related lines with and without betaine. Leaf tissue of both betaine-positive and betaine-deficient lines readily converted supplied betaine aldehyde to betaine, but only the betaine-containing line was able to oxidize supplied choline to betaine. This locates the lesion in betaine-deficient plants at the choline → betaine aldehyde step of betaine synthesis. Consistent with this location, betaine-deficient plants were shown to have no detectable endogenous pool of betaine aldehyde. PMID:16668098

  8. Maize Genetics and Genomics Database

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The 2007 report for MaizeGDB lists the new hires who will focus on curation/outreach and the genome sequence, respectively. Currently all sequence in the database comes from a PlantGDB pipeline and is presented with deep links to external resources such as PlantGDB, Dana Farber, GenBank, the Arizona...

  9. Ontogeny of the maize shoot apical meristem.

    PubMed

    Takacs, Elizabeth M; Li, Jie; Du, Chuanlong; Ponnala, Lalit; Janick-Buckner, Diane; Yu, Jianming; Muehlbauer, Gary J; Schnable, Patrick S; Timmermans, Marja C P; Sun, Qi; Nettleton, Dan; Scanlon, Michael J

    2012-08-01

    The maize (Zea mays) shoot apical meristem (SAM) arises early in embryogenesis and functions during stem cell maintenance and organogenesis to generate all the aboveground organs of the plant. Despite its integral role in maize shoot development, little is known about the molecular mechanisms of SAM initiation. Laser microdissection of apical domains from developing maize embryos and seedlings was combined with RNA sequencing for transcriptomic analyses of SAM ontogeny. Molecular markers of key events during maize embryogenesis are described, and comprehensive transcriptional data from six stages in maize shoot development are generated. Transcriptomic profiling before and after SAM initiation indicates that organogenesis precedes stem cell maintenance in maize; analyses of the first three lateral organs elaborated from maize embryos provides insight into their homology and to the identity of the single maize cotyledon. Compared with the newly initiated SAM, the mature SAM is enriched for transcripts that function in transcriptional regulation, hormonal signaling, and transport. Comparisons of shoot meristems initiating juvenile leaves, adult leaves, and husk leaves illustrate differences in phase-specific (juvenile versus adult) and meristem-specific (SAM versus lateral meristem) transcript accumulation during maize shoot development. This study provides insight into the molecular genetics of SAM initiation and function in maize.

  10. Influence of Different Supplements on the Commercial Cultivation of Milky White Mushroom

    PubMed Central

    Alam, Nuhu; Amin, Ruhul; Khair, Abul

    2010-01-01

    Calocybe indica, known as milky white mushroom, grows and cultivated in the sub-tropical and temperate zones of South Asia. We investigated the most suitable supplements and their levels for the commercial cultivation of milky white mushroom. Rice bran, maize powder, and wheat bran with their different levels (10, 20, 30, 40, and 50%) were used as supplements to evaluate the yield and yield contributing characteristics of C. indica. Primordia initiation was observed between 13.5 and 19.3 days. The results indicated that the 30% maize powder supplement was effective for producing viable fruiting bodies. The maximum diameters of the pileus and stalk were observed with 30% maize powder. The highest biological and economic yield and biological efficiency were also obtained with 30% maize powder as a supplement. The results indicate that increasing the supplement level resulted in less biological efficiency, and that 30% maize powder was the best supplement level for rice straw substrate to cultivate milky white mushrooms. PMID:23956652

  11. Influence of different supplements on the commercial cultivation of milky white mushroom.

    PubMed

    Alam, Nuhu; Amin, Ruhul; Khair, Abul; Lee, Tae Soo

    2010-09-01

    Calocybe indica, known as milky white mushroom, grows and cultivated in the sub-tropical and temperate zones of South Asia. We investigated the most suitable supplements and their levels for the commercial cultivation of milky white mushroom. Rice bran, maize powder, and wheat bran with their different levels (10, 20, 30, 40, and 50%) were used as supplements to evaluate the yield and yield contributing characteristics of C. indica. Primordia initiation was observed between 13.5 and 19.3 days. The results indicated that the 30% maize powder supplement was effective for producing viable fruiting bodies. The maximum diameters of the pileus and stalk were observed with 30% maize powder. The highest biological and economic yield and biological efficiency were also obtained with 30% maize powder as a supplement. The results indicate that increasing the supplement level resulted in less biological efficiency, and that 30% maize powder was the best supplement level for rice straw substrate to cultivate milky white mushrooms.

  12. Multi-mycotoxin analysis of maize silage by LC-MS/MS.

    PubMed

    Rasmussen, R R; Storm, I M L D; Rasmussen, P H; Smedsgaard, J; Nielsen, K F

    2010-05-01

    This paper describes a method for determination of 27 mycotoxins and other secondary metabolites in maize silage. The method focuses on analytes which are known to be produced by common maize and maize-silage contaminants. A simple pH-buffered sample extraction was developed on the basis of a very fast and simple method for analysis of multiple pesticide residues in food known as QuEChERS. The buffering effectively ensured a stable pH in samples of both well-ensiled maize (pH < 4) and of hot spots with fungal infection (pH > 7). No further clean-up was performed before analysis using liquid chromatography-tandem mass spectrometry. The method was successfully validated for determination of eight analytes qualitatively and 19 quantitatively. Matrix-matched calibration standards were used giving recoveries ranging from 37% to 201% with the majority between 60% and 115%. Repeatability (5-27% RSD(r)) and intra-laboratory reproducibility (7-35% RSD(IR)) was determined. The limit of detection (LOD) for the quantitatively validated analytes ranged from 1 to 739 microg kg(-1). Validation results for citrinin, fumonisin B(1) and fumonisin B(2) were unsatisfying. The method was applied to 20 selected silage samples and alternariol monomethyl ether, andrastin A, alternariol, citreoisocoumarin, deoxynivalenol, enniatin B, fumigaclavine A, gliotoxin, marcfortine A and B, mycophenolic acid, nivalenol, roquefortine A and C and zearalenone were detected.

  13. MaizeGDB: Global support for maize research through open access information [abstract

    Technology Transfer Automated Retrieval System (TEKTRAN)

    MaizeGDB is the open-access global repository for maize genetic and genomic information – from single genes that determine nutritional quality to whole genome-scale data for complex traits including yield and drought tolerance. The data and tools at MaizeGDB enable researchers from Ethiopia to Ghan...

  14. Maize genome sequencing by methylation filtration.

    PubMed

    Palmer, Lance E; Rabinowicz, Pablo D; O'Shaughnessy, Andrew L; Balija, Vivekanand S; Nascimento, Lidia U; Dike, Sujit; de la Bastide, Melissa; Martienssen, Robert A; McCombie, W Richard

    2003-12-19

    Gene enrichment strategies offer an alternative to sequencing large and repetitive genomes such as that of maize. We report the generation and analysis of nearly 100,000 undermethylated (or methylation filtration) maize sequences. Comparison with the rice genome reveals that methylation filtration results in a more comprehensive representation of maize genes than those that result from expressed sequence tags or transposon insertion sites sequences. About 7% of the repetitive DNA is unmethylated and thus selected in our libraries, but potentially active transposons and unmethylated organelle genomes can be identified. Reverse transcription polymerase chain reaction can be used to finish the maize transcriptome.

  15. Proteome Changes in Maize Embryo (Zea mays L) Induced by Ion Beam Implantation Treatment

    NASA Astrophysics Data System (ADS)

    Li, Yongliang; Tang, Jihua; Qin, Guangyong; Huo, Yuping; Tian, Shuangqi

    2009-08-01

    Low energy ion beam implantation was applied to the maize (Zea mays L) embryo proteome using two-dimensional gel electrophoresis. Protein profile analysis detected more than 1100 protein spots, 72 of which were determined to be expressed differently in the treated and control (not exposed to ion beam implantation) embryos. Of the 72 protein spots, 53 were up-regulated in the control and 19 were more abundantly expressed in the ion beam-treated embryos. The spots of up- or down-regulated proteins were identified by matrix assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF-MS). Among the identified proteins, 11 were up-regulated in the treated embryos. Four of these up-regulated proteins were antioxidant molecules, three were related to stress response, two to sugar metabolism and two were associated with heat shock response. Of the five proteins up-regulated in the control embryos, three were functionally related to carbohydrate metabolism; the functions of the remaining two proteins were unknown. The data collected during this study indicate that treatment of maize embryos with low energy ion beam implantation induces changes in stress tolerance enzymes/proteins, possibly as a result of alterations in metabolism.

  16. In vitro digestibility, crystallinity, rheological, thermal, particle size and morphological characteristics of pinole, a traditional energy food obtained from toasted ground maize.

    PubMed

    Carrera, Y; Utrilla-Coello, R; Bello-Pérez, A; Alvarez-Ramirez, J; Vernon-Carter, E J

    2015-06-05

    Flour obtained from toasted ground maize grains is widely consumed by different ethnic groups of Northern Mexico and Southwest USA as an energy source. In this work the in vitro digestibility, crystallinity, rheological, thermal, particle size distribution and morphological characteristics of toasted ground white and blue maize flours were studied. X-ray diffraction studies showed that the crystallinity content was reduced, but that the hydrolysis rate and the in vitro digestibility of starch were greatly improved by the toasting process. The relative amount of rapidly digestible starch showed an important increase at the expense of resistant starch content reduction. The thermal properties of white maize starch increased slightly, but those of the blue maize starch decreased slightly after toasting. Aqueous dispersions formed with 10% (w/w) flour were heated at 90°C for 5min to induce starch gelling, in order to resemble thin porridges. The dispersed gels exhibited higher elastic modulus (G') than loss modulus (G'') in the linear viscoelastic region, with blue maize dispersions displaying higher moduli magnitudes. At higher shear strain amplitudes, G' decreased but G'' first increased and then decreased (overshoot phenomenon). The effects of toasting on the structure and functionality of maize starch are explained on the basis of limited gelatinization of the granules. The results in this work provide insights for understanding the extensive use of pinole by impoverished ethnic groups, and more recently by high performance ultra-runners and athletes, as an energy food.

  17. Intermittency Models and Spot Measurements

    NASA Technical Reports Server (NTRS)

    Ashworth, D. A.

    2007-01-01

    Experimental work at the University of Oxford Osney Lab has demonstrated characteristics of the late-stage transition process by the use of thin-film heat transfer gauges. The development of turbulent spots has been observed in a range of environments, including flat plates, turbine blade cascade tests and wake-passing experiments. These results were taken at Mach/Reynolds numbers and gas-to-wall temperature ratios representative of gas turbines. Analyses of the spot characteristics are consistent with measurements taken in low speed experiments, and support the Schubauer and Klebanoff type of turbulent spots. The addition of simulated wakes from upstream stages has been observed to be primarily superpositional for these tests.

  18. Poisson's spot and Gouy phase

    NASA Astrophysics Data System (ADS)

    da Paz, I. G.; Soldati, Rodolfo; Cabral, L. A.; de Oliveira, J. G. G.; Sampaio, Marcos

    2016-12-01

    Recently there have been experimental results on Poisson spot matter-wave interferometry followed by theoretical models describing the relative importance of the wave and particle behaviors for the phenomenon. We propose an analytical theoretical model for Poisson's spot with matter waves based on the Babinet principle, in which we use the results for free propagation and single-slit diffraction. We take into account effects of loss of coherence and finite detection area using the propagator for a quantum particle interacting with an environment. We observe that the matter-wave Gouy phase plays a role in the existence of the central peak and thus corroborates the predominantly wavelike character of the Poisson's spot. Our model shows remarkable agreement with the experimental data for deuterium (D2) molecules.

  19. Laser based spot weld characterization

    NASA Astrophysics Data System (ADS)

    Jonietz, Florian; Myrach, Philipp; Rethmeier, Michael; Suwala, Hubert; Ziegler, Mathias

    2016-02-01

    Spot welding is one of the most important joining technologies, especially in the automotive industry. Hitherto, the quality of spot welded joints is tested mainly by random destructive tests. A nondestructive testing technique offers the benefit of cost reduction of the testing procedure and optimization of the fabrication process, because every joint could be examined. This would lead to a reduced number of spot welded joints, as redundancies could be avoided. In the procedure described here, the spot welded joint between two zinc-coated steel sheets (HX340LAD+Z100MB or HC340LA+ZE 50/50) is heated optically on one side. Laser radiation and flash light are used as heat sources. The melted zone, the so called "weld nugget" provides the mechanical stability of the connection, but also constitutes a thermal bridge between the sheets. Due to the better thermal contact, the spot welded joint reveals a thermal behavior different from the surrounding material, where the heat transfer between the two sheets is much lower. The difference in the transient thermal behavior is measured with time resolved thermography. Hence, the size of the thermal contact between the two sheets is determined, which is directly correlated to the size of the weld nugget, indicating the quality of the spot weld. The method performs well in transmission with laser radiation and flash light. With laser radiation, it works even in reflection geometry, thus offering the possibility of testing with just one-sided accessibility. By using heating with collimated laser radiation, not only contact-free, but also remote testing is feasible. A further convenience compared to similar thermographic approaches is the applicability on bare steel sheets without any optical coating for emissivity correction. For this purpose, a proper way of emissivity correction was established.

  20. [New technology in maize breeding].

    PubMed

    Konstantinov, K; Mladenović, S; Stojkov, S; Delić, N; Gosić, S; Petrović, R; Lević, J; Denić, M

    1992-01-01

    Results obtained by several approaches in the application of Biotechnology in maize breeding are reviewed. RFLP technology in the determination of genetic variation; gene transfer by the use of different methods of gene delivery and the determination of gene integration. Three technologies for foreign gene introduction have been applied; injection of plasmid pRT100 neo into archesporial tissue before micro and macro sporogenesis, slightly modified pollen-tube pathway technology and dry seed incubation in plasmid DNA solution. NPTII gene integration was followed by dot-blot and Southern blot analysis of plant DNA of both T1 and T2 plants. Gene expression was analysed by neomycin phosphotransferase activity. Transformed plants contained the selective NPTII gene sequence in an active form. Bacterial gene integration induced several heritable changes of plant phenotype. As an important change, alteration of the flowering time has been used as a criterion for selection and plant propagation to keep transformed progeny. Besides plant genome transformation, endogenous bacteria living in different maize tissue were found. As a perspective approach for biotechnology application in maize breeding biological vaccine construction has been selected. Therefore, antagonistic effect of gram positive bacterial strains to several pathogenic fungi was investigated. Results obtained after in vivo experiments are discussed.

  1. Resistance of maize landraces to the maize weevil Sitophilus zeamais Motsch. (Coleoptera: Curculionidae).

    PubMed

    Mikami, A Y; Carpentieri-Pípolo, V; Ventura, Maurício Ursi

    2012-10-01

    The maize weevil Sitophilus zeamais Motsch. is an important pest of maize that attacks the grain both in the field and during storage. The damage caused by the maize weevil S. zeamais on maize landraces, Amarelo Antigo, Asteca, Caiano, Carioca, and Ferrinho, was evaluated by no-choice tests under laboratory conditions. The commercial varieties Sol da Manhã, BR 106, BR 451, and the synthetics PC 0203 and PC 9903 were evaluated for comparisons with the maize landraces. The parameters evaluated were susceptibility index, number of weevil progeny, development time, weevil progeny dry weight, and grain dry weight loss. The landraces were more susceptible to the maize weevil as compared to the commercial varieties. Based on the cluster analysis, two groups of susceptibility to the maize weevil were observed: one of more susceptible populations formed by local landraces and BR 451, and another less susceptible, with commercial varieties, synthetics, and the landrace Amarelo.

  2. White Blood Cell Count

    MedlinePlus

    ... limited. Home Visit Global Sites Search Help? White Blood Cell Count Share this page: Was this page helpful? ... Count; Leukocyte Count; White Count Formal name: White Blood Cell Count Related tests: Complete Blood Count , Blood Smear , ...

  3. Rocky Mountain spotted fever, Panama.

    PubMed

    Estripeaut, Dora; Aramburú, María Gabriela; Sáez-Llorens, Xavier; Thompson, Herbert A; Dasch, Gregory A; Paddock, Christopher D; Zaki, Sherif; Eremeeva, Marina E

    2007-11-01

    We describe a fatal pediatric case of Rocky Mountain spotted fever in Panama, the first, to our knowledge, since the 1950s. Diagnosis was established by immunohistochemistry, PCR, and isolation of Rickettsia rickettsii from postmortem tissues. Molecular typing demonstrated strong relatedness of the isolate to strains of R. rickettsii from Central and South America.

  4. Registration of maize inbred line 'GT888'

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Maize (Zea mays L.) inbred line GT888 (PI 670116) was developed and released by the USDA-ARS in cooperation with the University of Georgia, and in participation with the USDA Germplasm Enhancement of Maize (GEM) project. GT888 was derived from GEM population DK888:N11 (GEMN-0177), which has 50% tro...

  5. Use of tropical maize for bioethanol production

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Tropical maize is an alternative energy crop being considered as a feedstock for bioethanol production in the North Central and Midwest United States. Tropical maize is advantageous because it produces large amounts of soluble sugars in its stalks, creates a large amount of biomass, and requires lo...

  6. A meteorologically driven maize stress indicator model

    NASA Technical Reports Server (NTRS)

    Taylor, T. W.; Ravet, F. W. (Principal Investigator)

    1981-01-01

    A maize soil moisture and temperature stress model is described which was developed to serve as a meteorological data filter to alert commodity analysts to potential stress conditions in the major maize-producing areas of the world. The model also identifies optimum climatic conditions and planting/harvest problems associated with poor tractability.

  7. Maize metabolic network construction and transcriptome analysis

    Technology Transfer Automated Retrieval System (TEKTRAN)

    A framework for understanding the synthesis and catalysis of metabolites and other biochemicals by proteins is crucial for unraveling the physiology of cells. To create such a framework for Zea mays ssp. mays (maize), we developed MaizeCyc a metabolic network of enzyme catalysts, proteins, carbohydr...

  8. The genetic architecture of maize height

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Height is one of the most heritable and easily measured traits in maize (Zea mays L.). Given a pedigree or estimates of the genomic identity-by-state (IBS) among related plants, height is also accurately predictable. But, mapping alleles explaining natural variation in maize height remains a formida...

  9. Viruses in maize and Johnsongrass in southern Ohio

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Two major maize viruses in the United States, Maize dwarf mosaic virus and Maize chlorotic dwarf virus, were first described in Southern Ohio and surrounding regions in the 1960s when they were major problems in maize (Zea mays L.) production. Planting resistant varieties and changing cultural prac...

  10. Pupillary light reflex to light inside the natural blind spot

    PubMed Central

    Miyamoto, Kentaro; Murakami, Ikuya

    2015-01-01

    When a light stimulus covers the human natural blind spot (BS), perceptual filling-in corrects for the missing information inside the BS. Here, we examined whether a filled-in surface of light perceived inside the BS affects the size of the short-latency pupillary light reflex (PLR), a pupil response mediated by a subcortical pathway for unconscious vision. The PLR was not induced by a red surface that was physically absent but perceptually filled-in inside the BS in the presence of a red ring surrounding it. However, a white large disk covering the BS unexpectedly induced a larger PLR than a white ring surrounding the BS border did, even though these two stimuli must be equivalent for the visual system, and trial-by-trial percepts did not predict PLR size. These results suggest that some physiological mechanism, presumably the retinal cells containing the photopigment melanopsin, receives the light projected inside the BS and enhances PLR. PMID:26115182

  11. 13C Natural Abundance of Serum Retinol Is a Novel Biomarker for Evaluating Provitamin A Carotenoid-Biofortified Maize Consumption in Male Mongolian Gerbils123

    PubMed Central

    Pungarcher, India; Mourao, Luciana; Davis, Christopher R; Simon, Philipp; Pixley, Kevin V; Tanumihardjo, Sherry A

    2016-01-01

    Background: Crops such as maize, sorghum, and millet are being biofortified with provitamin A carotenoids to ensure adequate vitamin A (VA) intakes. VA assessment can be challenging because serum retinol concentrations are homeostatically controlled and more sensitive techniques are resource-intensive. Objectives: We investigated changes in serum retinol relative differences of isotope amount ratios of 13C/12C (δ13C) caused by natural 13C fractionation in C3 compared with C4 plants as a biomarker to detect provitamin A efficacy from biofortified (orange) maize and high-carotene carrots. Methods: The design was a 2 × 2 × 2 maize (orange compared with white) by carrot (orange compared with white) by a VA fortificant (VA+ compared with VA−) in weanling male Mongolian gerbils (n = 55), which included a 14-d VA depletion period and a 62-d treatment period (1 baseline and 8 treatment groups; n = 5−7/group). Liver VA and serum retinol were quantified, purified by HPLC, and analyzed by GC combustion isotope ratio mass spectrometry for 13C. Results: Treatments affected liver VA concentrations (0.048 ± 0.039 to 0.79 ± 0.24 μmol/g; P < 0.0001) but not overall serum retinol concentrations (1.38 ± 0.22 μmol/L). Serum retinol and liver VA δ13C were significantly correlated (R2 = 0.92; P < 0.0001). Serum retinol δ13C differentiated control groups that consumed white maize and white carrots (−27.1 ± 1.2 δ13C‰) from treated groups that consumed orange maize and white carrots (−21.6 ± 1.4 δ13C‰ P < 0.0001) and white maize and orange carrots (−30.6 ± 0.7 δ13C‰ P < 0.0001). A prediction model demonstrated the relative contribution of orange maize to total dietary VA for groups that consumed VA from mixed sources. Conclusions: Provitamin A efficacy and quantitative estimation of the relative contribution to dietary VA were demonstrated with the use of serum retinol δ13C. This method could be used for maize efficacy or effectiveness studies and with

  12. Chemopreventive effects of free and bound phenolics associated to steep waters (nejayote) obtained after nixtamalization of different maize types.

    PubMed

    Rojas-García, Carlos; García-Lara, Silverio; Serna-Saldivar, Sergio O; Gutiérrez-Uribe, Janet A

    2012-03-01

    Free and bound phenolics extracts from nejayote solids were obtained after optimally lime-cooking blue, normal white, red, normal yellow, high-carotenoid and quality protein maize types. The extraction yield ranged from 4.47 to 10.05%. Bound phenolics extracts had higher content of total phenolics, antioxidant activity and ferulic acid compared to the free phenolics extracts. In general, free phenolics extracts were less cytotoxic than the bound phenolics counterparts. Bound phenolics extracts had higher induction of quinone reductase (QR) and particularly the normal yellow nejayote exerted the highest chemopreventive index tested in Hepa1c1c7 cells. When tested for monofunctional phase 2 induction capacity in BPrc1 cells, the bound phenolics extracts of blue, normal white and quality protein nejayotes were better inducers than the normal yellow counterpart. Particularly, the free phenolics extract of the white maize nejayote induced BPrc1 cells QR and exerted a higher chemopreventive index compared to the bound phenolics extract. Therefore, the nejayote of the normal white maize was the best source of monofunctional phase 2 enzyme inducers.

  13. White matter injury detection in neonatal MRI

    NASA Astrophysics Data System (ADS)

    Cheng, Irene; Hajari, Nasim; Firouzmanesh, Amirhossein; Shen, Rui; Miller, Steven; Poskitt, Ken; Basu, Anup

    2013-02-01

    Early detection of white matter injury in premature newborns can facilitate timely clinical treatments reducing the potential risk of later developmental deficits. It was reported that there were more than 5% premature newborns in British Columbia, Canada, among which 5-10% exhibited major motor deficits and 25-50% exhibited significant developmental and visual deficits. With the advancement of computer assisted detection systems, it is possible to automatically identify white matter injuries, which are found inside the grey matter region of the brain. Atlas registration has been suggested in the literature to distinguish grey matter from the soft tissues inside the skull. However, our subjects are premature newborns delivered at 24 to 32 weeks of gestation. During this period, the grey matter undergoes rapid changes and differs significantly from one to another. Besides, not all detected white spots represent injuries. Additional neighborhood information and expert input are required for verification. In this paper, we propose a white matter feature identification system for premature newborns, which is composed of several steps: (1) Candidate white matter segmentation; (2) Feature extraction from candidates; (3) Validation with data obtained at a later stage on the children; and (4) Feature confirmation for automated detection. The main challenge of this work lies in segmenting white matter injuries from noisy and low resolution data. Our approach integrates image fusion and contrast enhancement together with a fuzzy segmentation technique to achieve promising results. Other applications, such as brain tumor and intra-ventricular haemorrhage detection can also benefit from our approach.

  14. Expression characterization of genes for CMS-C in maize.

    PubMed

    Huang, Ling; Xiang, Jie; Liu, Jiazhou; Rong, Tingzhao; Wang, Jing; Lu, Yanli; Tang, Qilin; Wen, Wen; Cao, Moju

    2012-10-01

    Cytoplasmic male sterility (CMS)-C is one of the most attractive sources of male sterility in the production of hybrid maize. However, the abortion mechanism of CMS-C is currently unknown. The major aim of this work was to characterize the expression of genes and proteins during pollen abortion. The materials assayed included CMS-C line C48-2, its maintainer line N48-2, and fertile F(1) (C48-2 × 18 white). A total of 20 unique genes and 25 proteins were identified by suppression subtractive hybridization and 2-D electrophoresis, respectively. Most of the genes and proteins identified are closely related to energy metabolism, stress responses, molecular chaperones, and cell death, which are generally considered to be essential to pollen development. Based on the function of these identified genes and proteins, reactive oxygen species in isolated mitochondria and DNA fragments were analyzed. The results from this study indicate that the oxidative stress which was associated with the specific expression patterns of some genes may be the physiological cause for the abortion of premature microspores in the maize CMS-C line.

  15. Maize (Zea mays L.) seedling leaf nuclear proteome and differentially expressed proteins between a hybrid and its parental lines.

    PubMed

    Guo, Baojian; Chen, Yanhong; Li, Chuan; Wang, Tianya; Wang, Rui; Wang, Bo; Hu, Sha; Du, Xiaofen; Xing, Hongyan; Song, Xiao; Yao, Yingyin; Sun, Qixin; Ni, Zhongfu

    2014-05-01

    To better understand the underlying molecular basis of leaf development in maize, a reference map of nuclear proteins in basal region of seedling leaf was established using a combination of 2DE and MALDI-TOF-MS. In total, 441 reproducible protein spots in nuclear proteome of maize leaf basal region were detected with silver staining in a pH range of 3-10, among which 203 spots corresponding to 163 different proteins were identified. As expected, proteins implicated in RNA and protein-associated functions were overrepresented in nuclear proteome. Remarkably, a high percentage (10%) of proteins was identified to be involved in cell division and growth. In addition, comparative nuclear proteomic analysis in leaf basal region of highly heterotic hybrid Mo17/B73 and its parental lines was also performed and 52 of 445 (11.69%) detected protein spots were differentially expressed between the hybrid and its parental lines, among which 16 protein spots displayed nonadditively expressed pattern. These results indicated that hybridization between two parental lines can cause changes in the expression of a variety of nuclear proteins, which may be responsible for the observed leaf size heterosis.

  16. Transgenic maize plants by tissue electroporation.

    PubMed Central

    D'Halluin, K; Bonne, E; Bossut, M; De Beuckeleer, M; Leemans, J

    1992-01-01

    In this paper, we describe the transformation of regenerable maize tissues by electroporation. In many maize lines, immature zygotic embryos can give rise to embryogenic callus cultures from which plants can be regenerated. Immature zygotic embryos or embryogenic type I calli were wounded either enzymatically or mechanically and subsequently electroporated with a chimeric gene encoding neomycin phosphotransferase (neo). Transformed embryogenic calli were selected from electroporated tissues on kanamycin-containing media and fertile transgenic maize plants were regenerated. The neo gene was transmitted to the progeny of kanamycin-resistant transformants in a Mendelian fashion. This showed that all transformants were nonchimeric, suggesting that transformation and regeneration are a single-cell event. The maize transformation procedure presented here does not require the establishment of genotype-dependent embryogenic type II callus or cell suspension cultures and facilitates the engineering of new traits into agronomically relevant maize inbred lines. PMID:1334743

  17. Mutations in the maize zeta-carotene desaturase gene lead to viviparous kernel

    PubMed Central

    Chen, Yan; Li, Jiankun; Fan, Kaijian; Du, Yicong; Ren, Zhenjing; Xu, Jing; Zheng, Jun; Liu, Yunjun; Fu, Junjie; Ren, Dongtao

    2017-01-01

    Preharvest sprouting reduces the maize quality and causes a significant yield loss in maize production. vp-wl2 is a Mutator (Mu)-induced viviparous mutant in maize, causing white or pale yellow kernels, dramatically reduced carotenoid and ABA content, and a high level of zeta-carotene accumulation. Here, we reported the cloning of the vp-wl2 gene using a modified digestion-ligation-amplification method (DLA). The results showed that an insertion of Mu9 in the first intron of the zeta-carotene desaturase (ZDS) gene results in the vp-wl2 mutation. Previous studies have suggested that ZDS is likely the structural gene of the viviparous9 (vp9) locus. Therefore, we performed an allelic test using vp-wl2 and three vp9 mutants. The results showed that vp-wl2 is a novel allele of the vp9 locus. In addition, the sequences of ZDS gene were identified in these three vp9 alleles. The vp-wl2 mutant gene was subsequently introgressed into four maize inbred lines, and a viviparous phenotype was observed with yield losses from 7.69% to 13.33%. PMID:28339488

  18. Co-infection and disease severity of Ohio Maize dwarf mosaic virus and Maize chlorotic dwarf virus strains

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Two major maize viruses have been reported in the United States: Maize dwarf mosaic virus (MDMV) and Maize chlorotic dwarf virus (MCDV). These viruses co-occur in regions where maize is grown such that co-infections are likely. Co-infection of different strains of MCDV is also observed frequently...

  19. 9 CFR 149.4 - Spot audit.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... 9 Animals and Animal Products 1 2011-01-01 2011-01-01 false Spot audit. 149.4 Section 149.4... LIVESTOCK IMPROVEMENT VOLUNTARY TRICHINAE CERTIFICATION PROGRAM § 149.4 Spot audit. (a) In addition to regularly scheduled site audits, certified production sites will be subject to spot audits. (1) Random...

  20. 9 CFR 149.4 - Spot audit.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 9 Animals and Animal Products 1 2010-01-01 2010-01-01 false Spot audit. 149.4 Section 149.4... LIVESTOCK IMPROVEMENT VOLUNTARY TRICHINAE CERTIFICATION PROGRAM § 149.4 Spot audit. (a) In addition to regularly scheduled site audits, certified production sites will be subject to spot audits. (1) Random...

  1. White Men's Racial Others

    ERIC Educational Resources Information Center

    Lensmire, Timothy J.

    2014-01-01

    Background/Context: Increasingly, researchers and educators have argued that alternative conceptions of Whiteness and White racial identity are needed because current conceptions have undermined, rather than strengthened, our critical pedagogies with White people. Grounded in critical Whiteness studies, and drawing especially on the writings of…

  2. INTEGRATED WEED CONTROL IN MAIZE.

    PubMed

    Latré, J; Dewitte, K; Derycke, V; De Roo, B; Haesaert, G

    2015-01-01

    Integrated pest management has been implemented as a general practice by EU legislation. As weed control actually is the most important crop protection measure in maize for Western Europe, the new legislation will have its impact. The question is of course which systems can be successfully implemented in practice with respect to labour efficiency and economical parameters. During 3 successive growing seasons (2007, 2008, 2009) weed control in maize was evaluated, the main focus was put on different techniques of integrated weed control and was compared with chemical weed control. Additionally, during 4 successive growing seasons (2011, 2012, 2013 and 2014) two objects based on integrated weed control and two objects based on mechanical weed control were compared to about twenty different objects of conventional chemical weed control. One of the objects based on mechanical weed control consisted of treatment with the flex-tine harrow before and after emergence in combination with chemical weed control at a reduced rate in 3-4 leave stage. The second one consisted of broadcast mechanical treatments before and after emergence followed by a final in-row application of herbicides and an inter-row cultivation at 6-7(8) leave stage. All trials were conducted on the Experimental farm of Bottelare HoGent-UGent on a sandy loam soil. Maize was growing in 1/3 crop rotation. The effect on weed growth as well as the economic impact of the different applications was evaluated. Combining chemical and mechanical weed control is a possible option in conventional farming but the disadvantages must be taken into account. A better planned weed control based on the real present weed-population in combination with a carefully thought-out choice of herbicides should also be considered as an IPM--approach.

  3. Managing Rocky Mountain spotted fever.

    PubMed

    Minniear, Timothy D; Buckingham, Steven C

    2009-11-01

    Rocky Mountain spotted fever is caused by the tick-borne bacterium Rickettsia rickettsii. Symptoms range from moderate illness to severe illness, including cardiovascular compromise, coma and death. The disease is prevalent in most of the USA, especially during warmer months. The trademark presentation is fever and rash with a history of tick bite, although tick exposure is unappreciated in over a third of cases. Other signature symptoms include headache and abdominal pain. The antibiotic therapy of choice for R. rickettsii infection is doxycycline. Preventive measures for Rocky Mountain spotted fever and other tick-borne diseases include: wearing long-sleeved, light colored clothing; checking for tick attachment and removing attached ticks promptly; applying topical insect repellent; and treating clothing with permethrin.

  4. Identification of line-specific strategies for improving carotenoid production in synthetic maize through data-driven mathematical modeling.

    PubMed

    Comas, Jorge; Benfeitas, Rui; Vilaprinyo, Ester; Sorribas, Albert; Solsona, Francesc; Farré, Gemma; Berman, Judit; Zorrilla, Uxue; Capell, Teresa; Sandmann, Gerhard; Zhu, Changfu; Christou, Paul; Alves, Rui

    2016-09-01

    Plant synthetic biology is still in its infancy. However, synthetic biology approaches have been used to manipulate and improve the nutritional and health value of staple food crops such as rice, potato and maize. With current technologies, production yields of the synthetic nutrients are a result of trial and error, and systematic rational strategies to optimize those yields are still lacking. Here, we present a workflow that combines gene expression and quantitative metabolomics with mathematical modeling to identify strategies for increasing production yields of nutritionally important carotenoids in the seed endosperm synthesized through alternative biosynthetic pathways in synthetic lines of white maize, which is normally devoid of carotenoids. Quantitative metabolomics and gene expression data are used to create and fit parameters of mathematical models that are specific to four independent maize lines. Sensitivity analysis and simulation of each model is used to predict which gene activities should be further engineered in order to increase production yields for carotenoid accumulation in each line. Some of these predictions (e.g. increasing Zmlycb/Gllycb will increase accumulated β-carotenes) are valid across the four maize lines and consistent with experimental observations in other systems. Other predictions are line specific. The workflow is adaptable to any other biological system for which appropriate quantitative information is available. Furthermore, we validate some of the predictions using experimental data from additional synthetic maize lines for which no models were developed.

  5. Sweet Spots and Door Stops

    ERIC Educational Resources Information Center

    Thompson, Michael; Tsui, Stella; Leung, Chi Fan

    2011-01-01

    A sweet spot is referred to in sport as the perfect place to strike a ball with a racquet or bat. It is the point of contact between bat and ball where maximum results can be produced with minimal effort from the hand of the player. Similar physics can be applied to the less inspiring examples of door stops; the perfect position of a door stop is…

  6. Justifications shape ethical blind spots.

    PubMed

    Pittarello, Andrea; Leib, Margarita; Gordon-Hecker, Tom; Shalvi, Shaul

    2015-06-01

    To some extent, unethical behavior results from people's limited attention to ethical considerations, which results in an ethical blind spot. Here, we focus on the role of ambiguity in shaping people's ethical blind spots, which in turn lead to their ethical failures. We suggest that in ambiguous settings, individuals' attention shifts toward tempting information, which determines the magnitude of their lies. Employing a novel ambiguous-dice paradigm, we asked participants to report the outcome of the die roll appearing closest to the location of a previously presented fixation cross on a computer screen; this outcome would determine their pay. We varied the value of the die second closest to the fixation cross to be either higher (i.e., tempting) or lower (i.e., not tempting) than the die closest to the fixation cross. Results of two experiments revealed that in ambiguous settings, people's incorrect responses were self-serving. Tracking participants' eye movements demonstrated that people's ethical blind spots are shaped by increased attention toward tempting information.

  7. Systematic Propulsion Optimization Tools (SPOT)

    NASA Technical Reports Server (NTRS)

    Bower, Mark; Celestian, John

    1992-01-01

    This paper describes a computer program written by senior-level Mechanical Engineering students at the University of Alabama in Huntsville which is capable of optimizing user-defined delivery systems for carrying payloads into orbit. The custom propulsion system is designed by the user through the input of configuration, payload, and orbital parameters. The primary advantages of the software, called Systematic Propulsion Optimization Tools (SPOT), are a user-friendly interface and a modular FORTRAN 77 code designed for ease of modification. The optimization of variables in an orbital delivery system is of critical concern in the propulsion environment. The mass of the overall system must be minimized within the maximum stress, force, and pressure constraints. SPOT utilizes the Design Optimization Tools (DOT) program for the optimization techniques. The SPOT program is divided into a main program and five modules: aerodynamic losses, orbital parameters, liquid engines, solid engines, and nozzles. The program is designed to be upgraded easily and expanded to meet specific user needs. A user's manual and a programmer's manual are currently being developed to facilitate implementation and modification.

  8. Field and vegetable crops as hosts of larval western spotted cucumber beetle (Coleoptera: Chrysomelidae).

    PubMed

    Pedersen, Andrew B; Godfrey, Larry D

    2011-06-01

    The western spotted cucumber beetle, Diabrotica undecimpunctata undecimpunctata Mannerheim, is an important pest of melons (Cucurmis melo L.) in northern California. Recent observations indicate that adults are using alfalfa (Medicago sativa L.) as a feeding host and larvae may be developing on the roots. Greenhouse studies were conducted during the winters of 2009 and 2010 in which larval development was compared on the roots of six field and vegetable crops commonly grown in the southern Sacramento Valley. The growth parameters used to evaluate the hosts were larval and pupal head capsule width, body width, and body length as well as total survival percentage and survival percentage to the third instar. According to larval growth and survivorship in 2009, maize (Zea mays L.) was the best host, followed by alfalfa and tomato (Solanum lycopersicum L.) that were roughly equivalent to one another. Melon was a slightly weaker host than alfalfa and tomato; sunflower (Helianthus annuus L.) was a completely incompatible host and thus dropped from the 2010 study. In 2010, melon was the weakest host for larval development. Maize was the superior host again followed closely by alfalfa that performed slightly better than tomato and lima bean (Phaseolus lunatus L.). Data suggest that larval western spotted cucumber beetles may primarily develop outside of melon fields and the adults emigrate to melons.

  9. Effects of salts on the gelatinization and retrogradation properties of maize starch and waxy maize starch.

    PubMed

    Wang, Wei; Zhou, Hongxian; Yang, Hong; Zhao, Siming; Liu, Youming; Liu, Ru

    2017-01-01

    The objective of this study was to evaluate the effects of salts on the gelatinization and retrogradation of maize and waxy maize starch. Experimental results showed that the salting-out or structure-making ions, such as F(-) and SO4(2-), decreased the swelling power, solubility and transparency of both starches, but increased the gelatinization temperature, enthalpy, and syneresis, due to the tendency of these ions to protect the hydrogen bond links among starch molecules. On the other hand, the salting-in or structure-breaking ions, such as I(-) and SCN(-), exhibited the opposite effects. Microscopic observations confirmed such effects of salts on both starches. Furthermore, the effects of salts were more significant on waxy maize and on normal maize starch. Generally, salts could significantly influence on the gelatinization and retrogradation of maize and waxy maize starch, following the order of the Hofmeister series.

  10. Reshaping of the maize transcriptome by domestication

    PubMed Central

    Swanson-Wagner, Ruth; Briskine, Roman; Schaefer, Robert; Hufford, Matthew B.; Ross-Ibarra, Jeffrey; Myers, Chad L.; Tiffin, Peter; Springer, Nathan M.

    2012-01-01

    Through domestication, humans have substantially altered the morphology of Zea mays ssp. parviglumis (teosinte) into the currently recognizable maize. This system serves as a model for studying adaptation, genome evolution, and the genetics and evolution of complex traits. To examine how domestication has reshaped the transcriptome of maize seedlings, we used expression profiling of 18,242 genes for 38 diverse maize genotypes and 24 teosinte genotypes. We detected evidence for more than 600 genes having significantly different expression levels in maize compared with teosinte. Moreover, more than 1,100 genes showed significantly altered coexpression profiles, reflective of substantial rewiring of the transcriptome since domestication. The genes with altered expression show a significant enrichment for genes previously identified through population genetic analyses as likely targets of selection during maize domestication and improvement; 46 genes previously identified as putative targets of selection also exhibit altered expression levels and coexpression relationships. We also identified 45 genes with altered, primarily higher, expression in inbred relative to outcrossed teosinte. These genes are enriched for functions related to biotic stress and may reflect responses to the effects of inbreeding. This study not only documents alterations in the maize transcriptome following domestication, identifying several genes that may have contributed to the evolution of maize, but highlights the complementary information that can be gained by combining gene expression with population genetic analyses. PMID:22753482

  11. Cytological modifications in maize plants infected by barley yellow dwarf virus and maize dwarf mosaic virus.

    PubMed

    Musetti, R; Bruni, L; Favali, M A

    2002-01-01

    Three inbred lines of maize (33-16, MO17 and B73) differing in their susceptibility to Barley yellow dwarf virus and Maize dwarf mosaic virus were studied to compare the ultrastructural modifications induced by the two viruses in leaf tissues of different age. The results demonstrate that the alterations induced by the two viruses in the different maize lines could depend on the particular line tested.

  12. Downstream Development of a Laminar Spot

    NASA Astrophysics Data System (ADS)

    Sekiya, Naoki; Matsumoto, Akira

    It was well-known that a disturbance, introduced artificially into a supercritical laminar boundary layer along a flat plate, is still laminar in the initial stage of its downstream development. Thus, we named it a "laminar spot" because it resembles a turbulent spot though its velocity perturbation remains laminar. From velocity measurements using a rake-type 16-channel hot-wire probe, we found that in the first stage of the downstream development of a laminar spot, its maximum width was at 0.2δ (what is called the critical layer) and one-half of its lateral growth angle was about 5°, which is almost one-half that of a turbulent spot. We call this region a "laminar spot region". In the present study, we measured in detail the velocity field of a laminar spot using a new hot-wire probe in the laminar spot region. The results showed that a laminar spot consists of some hairpin vortices and some induced U-shaped vortices under the hairpin vortices. Because of the interaction of the velocities induced by the respective vortex legs, the legs of the U-shaped vortices were located at the outermost part of the spot. Moreover, the new vortex legs extended spanwise at about 4° as the spot traveled downstream. Consequently, we concluded that the laminar spot grew spanwise in accordance with the span of these vortex legs.

  13. Hardness methods for testing maize kernels.

    PubMed

    Fox, Glen; Manley, Marena

    2009-07-08

    Maize is a highly important crop to many countries around the world, through the sale of the maize crop to domestic processors and subsequent production of maize products and also provides a staple food to subsistance farms in undeveloped countries. In many countries, there have been long-term research efforts to develop a suitable hardness method that could assist the maize industry in improving efficiency in processing as well as possibly providing a quality specification for maize growers, which could attract a premium. This paper focuses specifically on hardness and reviews a number of methodologies as well as important biochemical aspects of maize that contribute to maize hardness used internationally. Numerous foods are produced from maize, and hardness has been described as having an impact on food quality. However, the basis of hardness and measurement of hardness are very general and would apply to any use of maize from any country. From the published literature, it would appear that one of the simpler methods used to measure hardness is a grinding step followed by a sieving step, using multiple sieve sizes. This would allow the range in hardness within a sample as well as average particle size and/or coarse/fine ratio to be calculated. Any of these parameters could easily be used as reference values for the development of near-infrared (NIR) spectroscopy calibrations. The development of precise NIR calibrations will provide an excellent tool for breeders, handlers, and processors to deliver specific cultivars in the case of growers and bulk loads in the case of handlers, thereby ensuring the most efficient use of maize by domestic and international processors. This paper also considers previous research describing the biochemical aspects of maize that have been related to maize hardness. Both starch and protein affect hardness, with most research focusing on the storage proteins (zeins). Both the content and composition of the zein fractions affect

  14. MaizeGDB: The Maize Model Organism Database for Basic, Translational, and Applied Research

    PubMed Central

    Lawrence, Carolyn J.; Harper, Lisa C.; Schaeffer, Mary L.; Sen, Taner Z.; Seigfried, Trent E.; Campbell, Darwin A.

    2008-01-01

    In 2001 maize became the number one production crop in the world with the Food and Agriculture Organization of the United Nations reporting over 614 million tonnes produced. Its success is due to the high productivity per acre in tandem with a wide variety of commercial uses. Not only is maize an excellent source of food, feed, and fuel, but also its by-products are used in the production of various commercial products. Maize's unparalleled success in agriculture stems from basic research, the outcomes of which drive breeding and product development. In order for basic, translational, and applied researchers to benefit from others' investigations, newly generated data must be made freely and easily accessible. MaizeGDB is the maize research community's central repository for genetics and genomics information. The overall goals of MaizeGDB are to facilitate access to the outcomes of maize research by integrating new maize data into the database and to support the maize research community by coordinating group activities. PMID:18769488

  15. Features of Jupiter's Great Red Spot

    NASA Technical Reports Server (NTRS)

    1996-01-01

    This montage features activity in the turbulent region of Jupiter's Great Red Spot (GRS). Four sets of images of the GRS were taken through various filters of the Galileo imaging system over an 11.5 hour period on 26 June, 1996 Universal Time. The sequence was designed to reveal cloud motions. The top and bottom frames on the left are of the same area, northeast of the GRS, viewed through the methane (732 nm) filter but about 70 minutes apart. The top left and top middle frames are of the same area and at the same time, but the top middle frame is taken at a wavelength (886 nm) where methane absorbs more strongly. (Only high clouds can reflect sunlight in this wavelength.) Brightness differences are caused by the different depths of features in the two images. The bottom middle frame shows reflected light at a wavelength (757 nm) where there are essentially no absorbers in the Jovian atmosphere. The white spot is to the northwest of the GRS; its appearance at different wavelengths suggests that the brightest elements are 30 km higher than the surrounding clouds. The top and bottom frames on the right, taken nine hours apart and in the violet (415 nm) filter, show the time evolution of an atmospheric wave northeast of the GRS. Visible crests in the top right frame are much less apparent 9 hours later in the bottom right frame. The misalignment of the north-south wave crests with the observed northwestward local wind may indicate a shift in wind direction (wind shear) with height. The areas within the dark lines are 'truth windows' or sections of the images which were transmitted to Earth using less data compression. Each of the six squares covers 4.8 degrees of latitude and longitude (about 6000 square kilometers). North is at the top of each frame.

    Launched in October 1989, Galileo entered orbit around Jupiter on December 7, 1995. The spacecraft's mission is to conduct detailed studies of the giant planet, its largest moons and the Jovian magnetic environment

  16. ESA uncovers Geminga's `hot spot'

    NASA Astrophysics Data System (ADS)

    2004-07-01

    16 July 2004 Astronomers using ESA’s X-ray observatory XMM-Newton have detected a small, bright ‘hot spot’ on the surface of the neutron star called Geminga, 500 light-years away. The hot spot is the size of a football field and is caused by the same mechanism producing Geminga’s X-ray tails. This discovery identifies the missing link between the X-ray and gamma-ray emission from Geminga. hi-res Size hi-res: 1284 kb Credits: ESA, P. Caraveo (IASF, Milan) Geminga's hot spot This figure shows the effects of charged particles accelerated in the magnetosphere of Geminga. Panel (a) shows an image taken with the EPIC instrument on board the XMM-Newton observatory. The bright tails, made of particles kicked out by Geminga’s strong magnetic field, trail the neutron star as it moves about in space. Panel (b) shows how electrically charged particles interact with Geminga’s magnetic field. For example, if electrons (blue) are kicked out by the star, positrons (in red) hit the star’s magnetic poles like in an ‘own goal’. Panel (c) illustrates the size of Geminga’s magnetic field (blue) compared to that of the star itself at the centre (purple). The magnetic field is tilted with respect to Geminga’s rotation axis (red). Panel (d) shows the magnetic poles of Geminga, where charged particles hit the surface of the star, creating a two-million degrees hot spot, a region much hotter than the surroundings. As the star spins on its rotation axis, the hot spot comes into view and then disappears, causing the periodic colour change seen by XMM-Newton. An animated version of the entire sequence can be found at: Click here for animated GIF [low resolution, animated GIF, 5536 KB] Click here for AVI [high resolution, AVI with DIVX compression, 19128 KB] hi-res Size hi-res: 371 kb Credits: ESA, P. Caraveo (IASF, Milan) Geminga's hot spot, panel (a) Panel (a) shows an image taken with the EPIC instrument on board the XMM-Newton observatory. The bright tails, made of

  17. Evaluation of Three Protein-Extraction Methods for Proteome Analysis of Maize Leaf Midrib, a Compound Tissue Rich in Sclerenchyma Cells

    PubMed Central

    Wang, Ning; Wu, Xiaolin; Ku, Lixia; Chen, Yanhui; Wang, Wei

    2016-01-01

    Leaf morphology is closely related to the growth and development of maize (Zea mays L.) plants and final kernel production. As an important part of the maize leaf, the midrib holds leaf blades in the aerial position for maximum sunlight capture. Leaf midribs of adult plants contain substantial sclerenchyma cells with heavily thickened and lignified secondary walls and have a high amount of phenolics, making protein extraction and proteome analysis difficult in leaf midrib tissue. In the present study, three protein-extraction methods that are commonly used in plant proteomics, i.e., phenol extraction, TCA/acetone extraction, and TCA/acetone/phenol extraction, were qualitatively and quantitatively evaluated based on 2DE maps and MS/MS analysis using the midribs of the 10th newly expanded leaves of maize plants. Microscopy revealed the existence of substantial amounts of sclerenchyma underneath maize midrib epidermises (particularly abaxial epidermises). The spot-number order obtained via 2DE mapping was as follows: phenol extraction (655) > TCA/acetone extraction (589) > TCA/acetone/phenol extraction (545). MS/MS analysis identified a total of 17 spots that exhibited 2-fold changes in abundance among the three methods (using phenol extraction as a control). Sixteen of the proteins identified were hydrophilic, with GRAVY values ranging from -0.026 to -0.487. For all three methods, we were able to obtain high-quality protein samples and good 2DE maps for the maize leaf midrib. However, phenol extraction produced a better 2DE map with greater resolution between spots, and TCA/acetone extraction produced higher protein yields. Thus, this paper includes a discussion regarding the possible reasons for differential protein extraction among the three methods. This study provides useful information that can be used to select suitable protein extraction methods for the proteome analysis of recalcitrant plant tissues that are rich in sclerenchyma cells. PMID:27379139

  18. Evaluation of Three Protein-Extraction Methods for Proteome Analysis of Maize Leaf Midrib, a Compound Tissue Rich in Sclerenchyma Cells.

    PubMed

    Wang, Ning; Wu, Xiaolin; Ku, Lixia; Chen, Yanhui; Wang, Wei

    2016-01-01

    Leaf morphology is closely related to the growth and development of maize (Zea mays L.) plants and final kernel production. As an important part of the maize leaf, the midrib holds leaf blades in the aerial position for maximum sunlight capture. Leaf midribs of adult plants contain substantial sclerenchyma cells with heavily thickened and lignified secondary walls and have a high amount of phenolics, making protein extraction and proteome analysis difficult in leaf midrib tissue. In the present study, three protein-extraction methods that are commonly used in plant proteomics, i.e., phenol extraction, TCA/acetone extraction, and TCA/acetone/phenol extraction, were qualitatively and quantitatively evaluated based on 2DE maps and MS/MS analysis using the midribs of the 10th newly expanded leaves of maize plants. Microscopy revealed the existence of substantial amounts of sclerenchyma underneath maize midrib epidermises (particularly abaxial epidermises). The spot-number order obtained via 2DE mapping was as follows: phenol extraction (655) > TCA/acetone extraction (589) > TCA/acetone/phenol extraction (545). MS/MS analysis identified a total of 17 spots that exhibited 2-fold changes in abundance among the three methods (using phenol extraction as a control). Sixteen of the proteins identified were hydrophilic, with GRAVY values ranging from -0.026 to -0.487. For all three methods, we were able to obtain high-quality protein samples and good 2DE maps for the maize leaf midrib. However, phenol extraction produced a better 2DE map with greater resolution between spots, and TCA/acetone extraction produced higher protein yields. Thus, this paper includes a discussion regarding the possible reasons for differential protein extraction among the three methods. This study provides useful information that can be used to select suitable protein extraction methods for the proteome analysis of recalcitrant plant tissues that are rich in sclerenchyma cells.

  19. Spatio-temporal dynamics of maize yield water constraints under climate change in Spain.

    PubMed

    Ferrero, Rosana; Lima, Mauricio; Gonzalez-Andujar, Jose Luis

    2014-01-01

    Many studies have analyzed the impact of climate change on crop productivity, but comparing the performance of water management systems has rarely been explored. Because water supply and crop demand in agro-systems may be affected by global climate change in shaping the spatial patterns of agricultural production, we should evaluate how and where irrigation practices are effective in mitigating climate change effects. Here we have constructed simple, general models, based on biological mechanisms and a theoretical framework, which could be useful in explaining and predicting crop productivity dynamics. We have studied maize in irrigated and rain-fed systems at a provincial scale, from 1996 to 2009 in Spain, one of the most prominent "hot-spots" in future climate change projections. Our new approach allowed us to: (1) evaluate new structural properties such as the stability of crop yield dynamics, (2) detect nonlinear responses to climate change (thresholds and discontinuities), challenging the usual linear way of thinking, and (3) examine spatial patterns of yield losses due to water constraints and identify clusters of provinces that have been negatively affected by warming. We have reduced the uncertainty associated with climate change impacts on maize productivity by improving the understanding of the relative contributions of individual factors and providing a better spatial comprehension of the key processes. We have identified water stress and water management systems as being key causes of the yield gap, and detected vulnerable regions where efforts in research and policy should be prioritized in order to increase maize productivity.

  20. Transcriptome Analysis of Maize Leaf Systemic Symptom Infected by Bipolaris zeicola

    PubMed Central

    Gong, Guoshu; Qin, Cheng; Ye, Kunhao; Zhang, Min; Sun, Xiaofang; Zhou, You; Zhang, Youju

    2015-01-01

    Bipolaris zeicola is a fungal pathogen that causes Northern corn leaf spot (NCLS), which is a serious foliar disease in maize and one of the most significant pathogens affecting global food security. Here, we report a genome-wide transcriptional profile analysis using next-generation sequencing (NGS) of maize leaf development after inoculation with B. zeicola. We performed High-Throughput Digital Gene Expression analysis to identify differentially expressed genes (DEGs) in resistant inbred Mo17 lines after infection with B. zeicola at four successive disease development stages—CP (contact period), PP (penetration period), IP (incubation period), and DP (disease period); the expression of the genes was compared with those in a CK (mock-treatment) control. In addition, a sensitive maize line (Zheng58) was used for the comparisons with the Mo17. Among all tested genes, 466 differentially expressed genes were identified in all libraries, and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis of these genes suggested that they are involved in many biological processes related to systemic symptom development, such as plant hormone signal transduction, starch and sucrose metabolism, phenylpropanoid biosynthesis and photosynthesis. Our systematic analysis provides comprehensive transcriptomic information regarding systemic symptom development in fungal-infected plants. This information will help in furthering our understanding of the detailed mechanisms of plant responses to fungal infection. PMID:25781606

  1. Performance of growing pigs fed diets based on by-products of maize and wheat processing.

    PubMed

    Mwesigwa, Robert; Mutetikka, David; Kugonza, Donald Rugira

    2013-02-01

    Forty-eight crossbred Landrace × Large white pigs with an average body weight of 8.5 ± 0.4 kg and aged 2 months were used to evaluate performance and carcass characteristics of pigs fed two diets in which bran from maize or wheat was used as the energy source. A third diet based on whole maize grain was used as a control. Animals in groups of four, balanced for litter, sex and weight were allotted to dietary treatments in a completely randomised design with four replications. Data were collected on feed intake and weight gain for a period of 4 months. A digestibility trial was carried at the end of the feeding trial using two male pigs per treatment while six pigs per treatment were randomly selected for slaughter to determine carcass characteristics. Daily gain averaged 0.23, 0.31 and 0.13 kg/day, for pigs fed maize bran, wheat bran and whole maize diets, respectively. Average feed intake and final body weight were significantly (P < 0.05) affected by diets. Digestibility of dry matter, crude fibre and calcium were higher (P < 0.05) for pigs fed maize bran while crude protein digestibility was highest (P < 0.05) for the pigs fed wheat bran. There were significant differences (P < 0.05) between the diets in carcass yield, ham, head, trotters and heart weights and also for rib weight, rib eye muscle and other tissues. This study indicates that comparable performance of growing pigs fed cereal bran and full-grain diets are real.

  2. Choosing a Genome Browser for a Model Organism Database (MOD): Surveying the Maize Community

    Technology Transfer Automated Retrieval System (TEKTRAN)

    As the maize genome sequencing is nearing its completion, the Maize Genetics and Genomics Database (MaizeGDB), the Model Organism Database for maize, integrated a genome browser to its already existing Web interface and database. The addition of the MaizeGDB Genome Browser to MaizeGDB will allow it ...

  3. Evaluation of maize yield in an on-farm maize-soybean and maize-Lablab crop rotation systems in the Northern Guinea Savanna of Nigeria.

    PubMed

    Okogun, J A; Sanginga, N; Abaidoo, R C

    2007-11-01

    An attempt was made to solving the problem of shortfall of fertilizer to maize production in the Northern Guinea Savanna (NGS) of Nigeria by harnessing the potentials of legume/cereal crop rotation in on-farm trials. The yield of maize that succeeded two soybean varieties and Lablab in a two-cycle of soybean/maize and Lablab/maize crop rotation in NGS Nigeria was assessed in researcher-managed and farmer-managed plots. Though maize that followed the soybean received between 5 kg N ha(-1) from improved soybean variety (TGx 1448-2E) and 17 kg N ha(-1) from farmer soybean variety (Samsoy-2) as N balance, this did not significantly (p = 0.05) affect the maize yields. The soybean shed 90-100% of its leaves at physiological maturity which resulted in about 110 kg N ha(-1) N uptake. This source of N might be one of the factors responsible for the increase in maize yield that followed soybean (20 to 24%) compared with continuous maize yield plot. Maize yield in previous Lablab plot was significantly (p = 0.05) higher than in all other treatments. Maize yield in farmer-managed plot ranged between 0.13 and 4.53 t ha(-1), maize yield in researcher-managed plot was over 200% higher than maize yield in farmer-managed plot because of poor crop management on the part of the farmer.

  4. β-Cryptoxanthin biofortified maize (Zea mays) increases β-cryptoxanthin concentration and enhances the color of chicken egg yolk.

    PubMed

    Liu, Y-Q; Davis, C R; Schmaelzle, S T; Rocheford, T; Cook, M E; Tanumihardjo, S A

    2012-02-01

    The laying hen has a natural ability to deposit carotenoids into its egg yolks, especially the xanthophyll carotenoid lutein that is used commercially as an egg colorant. Can this ability to deposit carotenoids be used to enrich egg yolk provitamin A value? After a 10-d carotenoid depletion period in hens (n = 24), the effects of a 20-d intervention with high-β-cryptoxanthin, high-β-carotene, or typical yellow maize on color and carotenoid profile were compared with the effects of a white maize diet (n = 6/treatment). Eggs were collected every other day and yolks were analyzed by using a portable colorimeter to define the color space and by using an HPLC to determine the carotenoid profile. The high-β-cryptoxanthin and yellow maize increased β-cryptoxanthin in the yolk (0.55 ± 0.08 to 4.20 ± 0.56 nmol/g and 0.55 ± 0.08 to 1.06 ± 0.12 nmol/g, respectively; P < 0.001). Provitamin A equivalents increased in eggs from hens fed high-β-cryptoxanthin maize (P < 0.001) but not the high-β-carotene maize. The color (L*, a*, and b*) assessment of the yolks showed an increase in the high-β-cryptoxanthin treatment for the red-green a* scale (P < 0.001) and a decrease for the light-dark L* scale (P < 0.001). No appreciable change was noted in the yellow-blue b* scale for the high-β-cryptoxanthin treatment; but significant changes were noted for the yellow (P = 0.002) and high-β-carotene maize (P = 0.005) treatments, which were most evident at the end of the washout period with white maize. β-Cryptoxanthin-biofortified maize is a potential vehicle to elevate provitamin A equivalents and to enhance the color of yolks. This could lead to a human health benefit if widely adopted.

  5. Cellulase from Trichoderma harzianum interacts with roots and triggers induced systemic resistance to foliar disease in maize

    PubMed Central

    Saravanakumar, Kandasamy; Fan, Lili; Fu, Kehe; Yu, Chuanjin; Wang, Meng; Xia, Hai; Sun, Jianan; Li, Yaqian; Chen, Jie

    2016-01-01

    Trichoderma harzianum is well known to exhibit induced systemic resistance (ISR) to Curvularia leaf spot. We previously reported that a C6 zinc finger protein (Thc6) is responsible for a major contribution to the ISR to the leaf disease, but the types of effectors and the signals mediated by Thc6 from Trichoderma are unclear. In this work, we demonstrated that two hydrolases, Thph1 and Thph2, from T. harzianum were regulated by Thc6. Furthermore, an electrophoretic mobility shift assay (EMSA) study revealed that Thc6 regulated mRNA expression by binding to GGCTAA and GGCTAAA in the promoters of the Thph1 and Thph2 genes, respectively. Moreover, the Thph1 and Thph2 proteins triggered the transient production of reactive oxygen species (ROS) and elevated the free cytosolic calcium levels in maize leaf. Furthermore, the genes related to the jasmonate/ethylene signaling pathway were up-regulated in the wild-type maize strain. However, the ΔThph1- or ΔThph2-deletion mutants could not activate the immune defense-related genes in maize to protect against leaf disease. Therefore, we conclude that functional Thph1 and Thph2 may be required in T. harzianum to activate ISR in maize. PMID:27830829

  6. Cellulase from Trichoderma harzianum interacts with roots and triggers induced systemic resistance to foliar disease in maize.

    PubMed

    Saravanakumar, Kandasamy; Fan, Lili; Fu, Kehe; Yu, Chuanjin; Wang, Meng; Xia, Hai; Sun, Jianan; Li, Yaqian; Chen, Jie

    2016-11-10

    Trichoderma harzianum is well known to exhibit induced systemic resistance (ISR) to Curvularia leaf spot. We previously reported that a C6 zinc finger protein (Thc6) is responsible for a major contribution to the ISR to the leaf disease, but the types of effectors and the signals mediated by Thc6 from Trichoderma are unclear. In this work, we demonstrated that two hydrolases, Thph1 and Thph2, from T. harzianum were regulated by Thc6. Furthermore, an electrophoretic mobility shift assay (EMSA) study revealed that Thc6 regulated mRNA expression by binding to GGCTAA and GGCTAAA in the promoters of the Thph1 and Thph2 genes, respectively. Moreover, the Thph1 and Thph2 proteins triggered the transient production of reactive oxygen species (ROS) and elevated the free cytosolic calcium levels in maize leaf. Furthermore, the genes related to the jasmonate/ethylene signaling pathway were up-regulated in the wild-type maize strain. However, the ΔThph1- or ΔThph2-deletion mutants could not activate the immune defense-related genes in maize to protect against leaf disease. Therefore, we conclude that functional Thph1 and Thph2 may be required in T. harzianum to activate ISR in maize.

  7. Watermarking spot colors in packaging

    NASA Astrophysics Data System (ADS)

    Reed, Alastair; Filler, TomáÅ.¡; Falkenstern, Kristyn; Bai, Yang

    2015-03-01

    In January 2014, Digimarc announced Digimarc® Barcode for the packaging industry to improve the check-out efficiency and customer experience for retailers. Digimarc Barcode is a machine readable code that carries the same information as a traditional Universal Product Code (UPC) and is introduced by adding a robust digital watermark to the package design. It is imperceptible to the human eye but can be read by a modern barcode scanner at the Point of Sale (POS) station. Compared to a traditional linear barcode, Digimarc Barcode covers the whole package with minimal impact on the graphic design. This significantly improves the Items per Minute (IPM) metric, which retailers use to track the checkout efficiency since it closely relates to their profitability. Increasing IPM by a few percent could lead to potential savings of millions of dollars for retailers, giving them a strong incentive to add the Digimarc Barcode to their packages. Testing performed by Digimarc showed increases in IPM of at least 33% using the Digimarc Barcode, compared to using a traditional barcode. A method of watermarking print ready image data used in the commercial packaging industry is described. A significant proportion of packages are printed using spot colors, therefore spot colors needs to be supported by an embedder for Digimarc Barcode. Digimarc Barcode supports the PANTONE spot color system, which is commonly used in the packaging industry. The Digimarc Barcode embedder allows a user to insert the UPC code in an image while minimizing perceptibility to the Human Visual System (HVS). The Digimarc Barcode is inserted in the printing ink domain, using an Adobe Photoshop plug-in as the last step before printing. Since Photoshop is an industry standard widely used by pre-press shops in the packaging industry, a Digimarc Barcode can be easily inserted and proofed.

  8. Open chromatin reveals the functional maize genome

    PubMed Central

    Rodgers-Melnick, Eli; Vera, Daniel L.; Bass, Hank W.

    2016-01-01

    Cellular processes mediated through nuclear DNA must contend with chromatin. Chromatin structural assays can efficiently integrate information across diverse regulatory elements, revealing the functional noncoding genome. In this study, we use a differential nuclease sensitivity assay based on micrococcal nuclease (MNase) digestion to discover open chromatin regions in the maize genome. We find that maize MNase-hypersensitive (MNase HS) regions localize around active genes and within recombination hotspots, focusing biased gene conversion at their flanks. Although MNase HS regions map to less than 1% of the genome, they consistently explain a remarkably large amount (∼40%) of heritable phenotypic variance in diverse complex traits. MNase HS regions are therefore on par with coding sequences as annotations that demarcate the functional parts of the maize genome. These results imply that less than 3% of the maize genome (coding and MNase HS regions) may give rise to the overwhelming majority of phenotypic variation, greatly narrowing the scope of the functional genome. PMID:27185945

  9. High-value products from transgenic maize.

    PubMed

    Naqvi, Shaista; Ramessar, Koreen; Farré, Gemma; Sabalza, Maite; Miralpeix, Bruna; Twyman, Richard M; Capell, Teresa; Zhu, Changfu; Christou, Paul

    2011-01-01

    Maize (also known as corn) is a domesticated cereal grain that has been grown as food and animal feed for tens of thousands of years. It is currently the most widely grown crop in the world, and is used not only for food/feed but also to produce ethanol, industrial starches and oils. Maize is now at the beginning of a new agricultural revolution, where the grains are used as factories to synthesize high-value molecules. In this article we look at the diversity of high-value products from maize, recent technological advances in the field and the emerging regulatory framework that governs how transgenic maize plants and their products are grown, used and traded.

  10. [Effects of nitrogen management on maize nitrogen utilization and residual nitrate nitrogen in soil under maize/soybean and maize/sweet potato relay strip intercropping systems].

    PubMed

    Wang, Xiao-Chun; Yang, Wen-Yu; Deng, Xiao-Yan; Zhang, Qun; Yong, Tai-Wen; Liu, Wei-Guo; Yang, Feng; Mao, Shu-Ming

    2014-10-01

    A large amount of nitrogen (N) fertilizers poured into the fields severely pollute the environment. Reasonable application of N fertilizer has always been the research hotpot. The effects of N management on maize N utilization and residual nitrate N in soil under maize/soybean and maize/ sweet potato relay strip intercropping systems were reported in a field experiment in southwest China. It was found that maize N accumulation, N harvest index, N absorption efficiency, N contribution proportion after the anthesis stage in maize/soybean relay strip intercropping were increased by 6.1%, 5.4%, 4.3%, and 15.1% than under maize/sweet potato with an increase of 22.6% for maize yield after sustainable growing of maize/soybean intercropping system. Nitrate N accumulation in the 0-60 cm soil layer was 12.9% higher under maize/soybean intercropping than under maize/sweet potato intercropping. However, nitrate N concentration in the 60-120 cm soil layer when intercropped with soybean decreased by 10.3% than when intercropped with sweet potato, indicating a decrease of N leaching loss. Increasing of N application rate enhanced N accumulation of maize and decreased N use efficiency and significantly increased nitrate concentration in the soil profile except in the 60-100 cm soil layer, where no significant difference was observed with nitrogen application rate at 0 to 270 kg · hm(-2). Further application of N fertilizer significantly enhanced nitrate leaching loss. Postponing N application increased nitrate accumulation in the 60-100 cm soil layer. The results suggested that N application rates and ratio of base to top dressing had different influences on maize N concentration and nitrate N between maize/soybean and maize/sweet potato intercropping. Maize N concentration in the late growing stage, N harvest index and N use efficiency under maize/soybean intercropping increased (with N application rate at 180-270 kg · hm(-2) and ratio of base to top dressing = 3:2:5) and

  11. Climate Change and Maize Yield in Iowa

    PubMed Central

    Xu, Hong; Twine, Tracy E.; Girvetz, Evan

    2016-01-01

    Climate is changing across the world, including the major maize-growing state of Iowa in the USA. To maintain crop yields, farmers will need a suite of adaptation strategies, and choice of strategy will depend on how the local to regional climate is expected to change. Here we predict how maize yield might change through the 21st century as compared with late 20th century yields across Iowa, USA, a region representing ideal climate and soils for maize production that contributes substantially to the global maize economy. To account for climate model uncertainty, we drive a dynamic ecosystem model with output from six climate models and two future climate forcing scenarios. Despite a wide range in the predicted amount of warming and change to summer precipitation, all simulations predict a decrease in maize yields from late 20th century to middle and late 21st century ranging from 15% to 50%. Linear regression of all models predicts a 6% state-averaged yield decrease for every 1°C increase in warm season average air temperature. When the influence of moisture stress on crop growth is removed from the model, yield decreases either remain the same or are reduced, depending on predicted changes in warm season precipitation. Our results suggest that even if maize were to receive all the water it needed, under the strongest climate forcing scenario yields will decline by 10–20% by the end of the 21st century. PMID:27219116

  12. Climate Change and Maize Yield in Iowa.

    PubMed

    Xu, Hong; Twine, Tracy E; Girvetz, Evan

    2016-01-01

    Climate is changing across the world, including the major maize-growing state of Iowa in the USA. To maintain crop yields, farmers will need a suite of adaptation strategies, and choice of strategy will depend on how the local to regional climate is expected to change. Here we predict how maize yield might change through the 21st century as compared with late 20th century yields across Iowa, USA, a region representing ideal climate and soils for maize production that contributes substantially to the global maize economy. To account for climate model uncertainty, we drive a dynamic ecosystem model with output from six climate models and two future climate forcing scenarios. Despite a wide range in the predicted amount of warming and change to summer precipitation, all simulations predict a decrease in maize yields from late 20th century to middle and late 21st century ranging from 15% to 50%. Linear regression of all models predicts a 6% state-averaged yield decrease for every 1°C increase in warm season average air temperature. When the influence of moisture stress on crop growth is removed from the model, yield decreases either remain the same or are reduced, depending on predicted changes in warm season precipitation. Our results suggest that even if maize were to receive all the water it needed, under the strongest climate forcing scenario yields will decline by 10-20% by the end of the 21st century.

  13. Nonbright spot AVO: Two examples

    SciTech Connect

    Ross, C.P.; Kinman, D.L.

    1994-12-31

    Utilization of amplitude-versus-offset (AVO) attribute sections such as the product of the normal incidence trace and gradient trace have been used extensively in bright spot (Class 3) AVO analysis and interpretation. However, while these sections have often worked well with Class 3 responses they are not reliable indicators of non-bright spot (Class 2) seismic anomalies. Analyzing Class 2 seismic data with AVO products will: (1) not detect the gas-charged reservoir because of near-zero acoustic impedance contrast between the sands and encasing shales, or (2) yield an incorrect (negative) AVO product if the normal incidence and gradient values are opposite in sign. Class 2 offset responses are divided into two sub-categories: those with phase reversals (Class 2p) and those without phase reversals (Class 2). An AVO procedure for these types of Class 2 anomalies is presented through two examples. The technique better exploits the nature of the prestack response, yielding a more definitive AVO attribute section, and the technique is adaptive to both Class 2 and Class 2p responses. When compared to a conventionally processed relative amplitude seismic section with characteristically low amplitude responses for near-zero acoustic impedance sands, this procedure clearly denotes the presence of gas-charged pore fluids within the reservoir.

  14. Spot and Runway Departure Advisor

    NASA Technical Reports Server (NTRS)

    Jung, Yoon Chul

    2013-01-01

    The Spot and Runway Departure Advisor (SARDA) is a research prototype of a decision support tool for ATC tower controllers to assist in manging and controlling traffic on the surface of an airport. SARDA employs a scheduler to generate an optimal runway schedule and gate push-back - spot release sequence and schedule that improves efficiency of surface operations. The advisories for ATC tower controllers are displayed on an Electronic Flight Strip (EFS) system. The human-in-the-loop simulation of the SARDA tool was conducted for east operations of Dallas-Ft. Worth International Airport (DFW) to evaluate performance of the SARDA tool and human factors, such as situational awareness and workload. The results indicates noticeable taxi delay reduction and fuel savings by using the SARDA tool. Reduction in controller workload were also observed throughout the scenario runs. The future plan includes modeling and simulation of the ramp operations of the Charlotte International Airport, and develop a decision support tool for the ramp controllers.

  15. Instructor Debrief Training in SPOT

    NASA Technical Reports Server (NTRS)

    Martin, Lynne; Orasanu, Judith; Villeda, Eric; Conners, Mary M. (Technical Monitor)

    2002-01-01

    One way to enhance the effectiveness of Special Purpose Operational Training' (SPOT) debriefing sessions may be for instructors to make explicit connections between the Crew Resource Management (CRM) concepts a carrier advocates and the behaviors displayed by the crew in question. A tool listing key behaviors from the scenario was devised, accompanied by an instructors' training session in which links were made between the behaviors and the underlying CRM processes they reflect. The aim of the tool is to assist instructors to focus the debriefing on the key SPOT/ CRM issues, in this case on planning. A second tool suggested ways to facilitate the discussion. Fourteen instructors at a major U.S. carrier took part in the training session and used the toolkit in their subsequent debriefs. Pre- and post-training debriefing samples from each instructor were compared to assess whether there were any changes in instructors' approaches to discussions in terms of the topics they covered and how they raised the points.

  16. MaizeGDB: The Maize Model Organism Database for Basic, Translational, and Applied Research

    Technology Transfer Automated Retrieval System (TEKTRAN)

    In 2001, maize became the number one production crop in the world (with over 614 million tons produced; http://faostat.fao.org). Its success is due to the high productivity per acre in tandem with a wide variety of commercial uses: not only is maize an excellent source of food, feed, and fuel, its...

  17. MaizeGDB update: New tools, data, and interface for the maize model organism database

    Technology Transfer Automated Retrieval System (TEKTRAN)

    MaizeGDB is a highly curated, community-oriented database and informatics service to researchers focused on the crop plant and model organism Zea mays ssp. mays. Although some form of the maize community database has existed over the last 25 years, there have only been two major releases. In 1991, ...

  18. Impact of 9 years of Bt-maize cultivation on the distribution of maize viruses.

    PubMed

    Achon, Maria Angeles; Alonso-Dueñas, Natalia

    2009-06-01

    This study assesses the effect of Bt-maize on the distribution of maize viruses. Random surveys were conducted in Spain between 2001 and 2006 to evaluate the occurrence of maize viruses in Bt-maize cultivation areas and in areas where this crop had not been introduced. Maize dwarf mosaic virus (MDMV) was the predominant virus in Bt-areas, and Maize rough dwarf virus (MRDV) was the most predominant one in non-Bt-areas, with MRDV an emergent virus in both types of areas. A decline in the occurrence of MDMV and an increase in that of Sugarcane mosaic virus was observed in Bt-areas. Additionally, data obtained over 6 years in experimental fields showed non-significant differences between the infection rates exhibited by two generations of Bt varieties and the non-transformed isogenics varieties for any of the viruses. Our data suggest that differences in virus distribution are linked to the genetic background of the maize varieties and the distribution of virus reservoirs rather than to Bt-maize cultivation.

  19. Expression of an anthranilate synthase from maize mutant bf-1 in maize line HiII

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Maize mutant bf-1 was one of a series of maize mutants generated by radiation from the Bikini Atoll atomic bomb test in 1946. It is characterized by blue fluorescence in seedlings and anthers under ultraviolet illumination and by mutant plants giving off a characteristic grape-like odor due to the ...

  20. Identification of resistance to Maize rayado fino virus in maize inbred lines

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Maize rayado fino virus (MRFV) is one of the most important virus diseases of maize in America. Severe yield losses, ranging from 10 to 50% in landraces to nearly 100% in contemporary cultivars, have been reported. Resistance has been reported in populations, but few inbred lines have been identifie...

  1. Expanding maize genetic resources with predomestication alleles: maize-teosinte introgression populations

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Teosinte (Zea mays ssp. parviglumis) has greater genetic diversity than maize inbreds and landraces (Z. mays ssp. mays). There are, however, limited genetic resources to efficiently evaluate and tap this diversity. To broaden resources for genetic diversity studies in maize, we developed and evaluat...

  2. Network analysis of maize RNA transport pathway genes associated with maize resistance to aflatoxin accumulation

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Aspergillus flavus is a pathogenic fungus producing alfatoxins that cause significant economic losses in maize production. This study analyzes the differences in expression levels of maize genes in response to A. flavus infection and aflatoxin accumulation. Identification of defense related genes an...

  3. Solar 'hot spots' are still hot

    NASA Technical Reports Server (NTRS)

    Bai, Taeil

    1990-01-01

    Longitude distributions of solar flares are not random but show evidence for active zones (or hot spots) where flares are concentrated. According to a previous study, two hot spots in the northern hemisphere, which rotate with a synodic period of about 26.72 days, produced the majority of major flares, during solar cycles 20 and 21. The more prominent of these two hot spots is found to be still active during the rising part of cycle 22, producing the majority of northern hemisphere major flares. The synodic rotation period of this hot spot is 26.727 + or - 0.007 days. There is also evidence for hot spots in the southern hemisphere. Two hot spots separated by 180 deg are found to rotate with a period of 29.407 days, with one of them having persisted in the same locations during cycles 19-22 and the other, during cycles 20-22.

  4. Spotted bass Micropterus punctulatus (Rafinesque 1819)

    USGS Publications Warehouse

    Churchill, Timothy N.; Bettoli, Phillip William

    2015-01-01

    Three subspecies of Spotted Bass Micropterus punctulatus were historically recognized: the smaller Northern Spotted Bass M. p. punctulatus, the larger, longer-lived Alabama Spotted Bass M. p. henshalli, and the now invalidated Wichita Spotted Bass M. p. wichitae (Bailey and Hubbs 1940; Cofer 1995; Warren 2009; Rider and Maceina 2015, this volume). The subspecific status has been examined over the past decade as advanced genetic analyses have been developed (e.g., Kassler et al. 2002; Baker et al. 2008; Tringali et al. 2015, this volume). The American Fisheries Society has recently changed the designation of the Alabama Spotted Bass to a separate species, Alabama Bass M. henshalli (Page et al. 2013). The remainder of this paper will discuss the biology and conservation of only Spotted Bass. Both species have been observed to hybridize with other Micropterus spp. (Koppelman 1994; Pierce and Van Den Avyle 1997; Barwick et al. 2006).

  5. Integrating sustainable hunting in biodiversity protection in Central Africa: hot spots, weak spots, and strong spots.

    PubMed

    Fa, John E; Olivero, Jesús; Farfán, Miguel Ángel; Márquez, Ana Luz; Vargas, Juan Mario; Real, Raimundo; Nasi, Robert

    2014-01-01

    Wild animals are a primary source of protein (bushmeat) for people living in or near tropical forests. Ideally, the effect of bushmeat harvests should be monitored closely by making regular estimates of offtake rate and size of stock available for exploitation. However, in practice, this is possible in very few situations because it requires both of these aspects to be readily measurable, and even in the best case, entails very considerable time and effort. As alternative, in this study, we use high-resolution, environmental favorability models for terrestrial mammals (N = 165) in Central Africa to map areas of high species richness (hot spots) and hunting susceptibility. Favorability models distinguish localities with environmental conditions that favor the species' existence from those with detrimental characteristics for its presence. We develop an index for assessing Potential Hunting Sustainability (PHS) of each species based on their ecological characteristics (population density, habitat breadth, rarity and vulnerability), weighted according to restrictive and permissive assumptions of how species' characteristics are combined. Species are classified into five main hunting sustainability classes using fuzzy logic. Using the accumulated favorability values of all species, and their PHS values, we finally identify weak spots, defined as high diversity regions of especial hunting vulnerability for wildlife, as well as strong spots, defined as high diversity areas of high hunting sustainability potential. Our study uses relatively simple models that employ easily obtainable data of a species' ecological characteristics to assess the impacts of hunting in tropical regions. It provides information for management by charting the geography of where species are more or less likely to be at risk of extinction from hunting.

  6. Concentration and dissipation of chlorantraniliprole and thiamethoxam residues in maize straw, maize, and soil.

    PubMed

    He, Min; Song, Dan; Jia, Hong C; Zheng, Yongquan

    2016-09-01

    To study the dissipation rates and final residual levels of chlorantraniliprole and thiamethoxam in maize straw, maize, and soil, two independent field trials were conducted during the 2014 cropping season in Beijing and Anhui Provinces of China. A 40% wettable powder (20% chlorantraniliprole + 20% thiamethoxam) was sprayed onto maize straw and soil at an application rate of 118 g of active ingredient per hectare (g a.i.ha(-1)). The residual concentrations were determined by ultra-high-performance liquid chromatography-tandem mass spectrometry. The chlorantraniliprole half-lives in maize straw and soil were 9.0-10.8 and 9.5-21.7 days, respectively. The thiamethoxam half-lives in maize straw and soil were 8.4-9.8 and 4.3-11.7 days, respectively. The final residues of chlorantraniliprole and thiamethoxam in maize straw, maize, and soil were measured after the pesticides had been sprayed two and three times with an interval of 7 days using 1 and 1.5 times the recommended rate (72 g a.i. ha(-1) and 108 g a.i. ha(-1), respectively). Representative maize straw, maize, and soil samples were collected after the last treatment at pre-harvest intervals of 7, 14, and 28 days. The chlorantraniliprole residue was below 0.01 mg kg(-1) in maize, between 0.01 and 0.31 mg kg(-1) in maize straw, and between 0.03 and 1.91 mg kg(-1) in soil. The thiamethoxam residue concentrations in maize, maize straw, and soil were <0.01, <0.01, and 0.01-0.03 mg kg(-1), respectively. The final pesticide residues on maize were lower than the maximum residue limit (MRL) of 0.02 mg kg(-1) after a 14-day pre-harvest interval. Therefore, a dosage of 72 g a.i. ha(-1) was recommended, as it can be considered safe to human beings and animals.

  7. Confronting White Privilege

    ERIC Educational Resources Information Center

    Swalwell, Katy

    2012-01-01

    Even as the United States becomes more diverse, a new era of "white flight" is unfolding. Whether they live in urban, suburban or rural communities, white students are likely to attend schools that reinforce their perceptions of cultural dominance. The average white student attends a school where 77 percent of the student body is of their race.…

  8. Leaf transpiration efficiency of some drought-resistant maize lines

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Field measurements of leaf gas exchange in maize often indicate stomatal conductances higher than required to provide substomatal carbon dioxide concentrations saturating to photosynthesis. Thus maize leaves often operate at lower transpiration efficiency (TE) than potentially achievable for specie...

  9. Genetic architecture of domestication-related traits in maize

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Strong directional selection occurred during the domestication of maize from its wild ancestor teosinte, reducing its genetic diversity, particularly at genes controlling domestication-related traits. Nevertheless, variability for some domestication-related traits is maintained in maize. The genet...

  10. Comparison of immune response of Pacific white shrimp, Litopenaeus vannamei, after multiple and single infections with WSSV and Vibrio anguillarum

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Our previous study demonstrated that Pacific white shrimp (Litopenaeus vannamei) infected by multiple pathogens showed higher mortality and death occurred more quickly than those infected by a single pathogen [1]. For better understanding the defense mechanism against white spot syndrome virus (WSSV...

  11. Study Progress on Tissue Culture of Maize Mature Embryo

    NASA Astrophysics Data System (ADS)

    Wang, Hongzhen; Cheng, Jun; Cheng, Yanping; Zhou, Xioafu

    It has been paid more and more attention on maize tissue culture as it is a basic work in maize genetic transformation, especially huge breakthrough has been made in maize tissue culture utilizing mature embryos as explants in the recent years. This paper reviewed the study progress on maize tissue culture and plant regeneration utilizing mature embryos as explants from callus induction, subculture, plant regeneration and browning reduction and so on.

  12. The genetic architecture of maize height.

    PubMed

    Peiffer, Jason A; Romay, Maria C; Gore, Michael A; Flint-Garcia, Sherry A; Zhang, Zhiwu; Millard, Mark J; Gardner, Candice A C; McMullen, Michael D; Holland, James B; Bradbury, Peter J; Buckler, Edward S

    2014-04-01

    Height is one of the most heritable and easily measured traits in maize (Zea mays L.). Given a pedigree or estimates of the genomic identity-by-state among related plants, height is also accurately predictable. But, mapping alleles explaining natural variation in maize height remains a formidable challenge. To address this challenge, we measured the plant height, ear height, flowering time, and node counts of plants grown in >64,500 plots across 13 environments. These plots contained >7300 inbreds representing most publically available maize inbreds in the United States and families of the maize Nested Association Mapping (NAM) panel. Joint-linkage mapping of quantitative trait loci (QTL), fine mapping in near isogenic lines (NILs), genome-wide association studies (GWAS), and genomic best linear unbiased prediction (GBLUP) were performed. The heritability of maize height was estimated to be >90%. Mapping NAM family-nested QTL revealed the largest explained 2.1 ± 0.9% of height variation. The effects of two tropical alleles at this QTL were independently validated by fine mapping in NIL families. Several significant associations found by GWAS colocalized with established height loci, including brassinosteroid-deficient dwarf1, dwarf plant1, and semi-dwarf2. GBLUP explained >80% of height variation in the panels and outperformed bootstrap aggregation of family-nested QTL models in evaluations of prediction accuracy. These results revealed maize height was under strong genetic control and had a highly polygenic genetic architecture. They also showed that multiple models of genetic architecture differing in polygenicity and effect sizes can plausibly explain a population's variation in maize height, but they may vary in predictive efficacy.

  13. Voyager 1 Red Spot Movie

    NASA Technical Reports Server (NTRS)

    2000-01-01

    This movie shows the portion of Jupiter around the Great Red Spot as it swirls through more than 60 Jupiter days. Notice the difference in speed and direction of the various zones of the atmosphere. The interaction of the atmospheric clouds and storm shows how dynamic the Jovian atmosphere is.

    As Voyager 1 approached Jupiter in 1979, it took images of the planet at regular intervals. This sequence is made from 66 images taken once every Jupiter rotation period (about 10 hours). This time-lapse movie uses images taken every time Jupiter longitude 68W passed under the spacecraft. These images were acquired in the Blue filter from Jan. 6 to Feb. 3 1979. The spacecraft flew from 58 million kilometers to 31 million kilometers from Jupiter during that time.

    This time-lapse movie was produced at JPL by the Image Processing Laboratory in 1979.

  14. Fatal Rocky Mountain spotted fever.

    PubMed

    Hattwick, M A; Retailliau, H; O'Brien, R J; Slutzker, M; Fontaine, R E; Hanson, B

    1978-09-29

    Forty-four fatal cases of Rocky Mountain spotted fever (RMSF) occurring in 1974 were compared with 50 nonfatal cases of similar age, sex, date of onset, and place of occurrence. Diagnosis and initiation of treatment in fatal cases were substantially delayed compared with nonfatal cases. Several reasons for this delay were identified: (1) the rash appeared later in the course of illness in the fatal cases, often not until the patient was terminal, (2) a history of tick bite was less often obtained during life or obtained late in the clinical course in fatal cases, and (3) initial nonspecific symptoms or unexpected symptoms led to an initial diagnosis of more common diseases. Only two fatal cases were treated with either tetracycline or chloramphenicol before the sixth day of illness. Presumptive diagnosis of RMSF and initiation of tetracycline therapy before onset of rash may be necessary to reduce mortality.

  15. SU-E-T-510: Interplay Between Spots Sizes, Spot / Line Spacing and Motion in Spot Scanning Proton Therapy

    SciTech Connect

    Lee, TK

    2015-06-15

    Purpose In proton beam configuration for spot scanning proton therapy (SSPT), one can define the spacing between spots and lines of scanning as a ratio of given spot size. If the spacing increases, the number of spots decreases which can potentially decrease scan time, and so can whole treatment time, and vice versa. However, if the spacing is too large, the uniformity of scanned field decreases. Also, the field uniformity can be affected by motion during SSPT beam delivery. In the present study, the interplay between spot/ line spacing and motion is investigated. Methods We used four Gaussian-shape spot sizes with 0.5cm, 1.0cm, 1.5cm, and 2.0cm FWHM, three spot/line spacing that creates uniform field profile which are 1/3*FWHM, σ/3*FWHM and 2/3*FWHM, and three random motion amplitudes within, +/−0.3mm, +/−0.5mm, and +/−1.0mm. We planned with 2Gy uniform single layer of 10×10cm2 and 20×20cm2 fields. Then, mean dose within 80% area of given field size, contrubuting MU per each spot assuming 1cGy/MU calibration for all spot sizes, number of spots and uniformity were calculated. Results The plans with spot/line spacing equal to or smaller than 2/3*FWHM without motion create ∼100% uniformity. However, it was found that the uniformity decreases with increased spacing, and it is more pronounced with smaller spot sizes, but is not affected by scanned field sizes. Conclusion It was found that the motion during proton beam delivery can alter the dose uniformity and the amount of alteration changes with spot size which changes with energy and spot/line spacing. Currently, robust evaluation in TPS (e.g. Eclipse system) performs range uncertainty evaluation using isocenter shift and CT calibration error. Based on presented study, it is recommended to add interplay effect evaluation to robust evaluation process. For future study, the additional interplay between the energy layers and motion is expected to present volumetric effect.

  16. New Observations of the Mysterious Metal-Polluted White Dwarf GD 394

    NASA Astrophysics Data System (ADS)

    Wilson, D. J.; Gänsicke, B. T.; Koester, D.; Holberg, J.; Burleigh, M. R.; Belardi, C.

    2017-03-01

    GD394 is a hot, extremely metal-polluted white dwarf. EUVE observations in the early 1990s revealed a 1.15 day periodicity with a 25 percent amplitude. This was hypothesised to be due to an accretion spot on the white dwarf, rotating into and out of view on the white dwarf spin period. We have obtained phase-resolved HST/STIS high-resolution FUV spectra of GD394 that sample the entire spin period. The spectra contain dozens of strong metal absorption lines, which we use to test the hypothesis of a rotating spot. No variation is seen in the STIS spectra, nor between them and a 1992 GHRS spectrum. Furthermore, archive SuperWASP observations do not show the optical variability predicted by the spot hypothesis. The STIS spectra also show mysterious C IV lines that may be an indicator of circumstellar material.

  17. Real-time quantitative polymerase chain reaction methods for four genetically modified maize varieties and maize DNA content in food.

    PubMed

    Brodmann, Peter D; Ilg, Evelyn C; Berthoud, Hélène; Herrmann, Andre

    2002-01-01

    Quantitative detection methods are needed for enforcement of the recently introduced labeling threshold for genetically modified organisms (GMOs) in food ingredients. This labeling threshold, which is set to 1% in the European Union and Switzerland, must be applied to all approved GMOs. Four different varieties of maize are approved in the European Union: the insect-resistant Bt176 maize (Maximizer), Btl 1 maize, Mon810 (YieldGard) maize, and the herbicide-tolerant T25 (Liberty Link) maize. Because the labeling must be considered individually for each ingredient, a quantitation system for the endogenous maize content is needed in addition to the GMO-specific detection systems. Quantitative real-time polymerase chain reaction detection methods were developed for the 4 approved genetically modified maize varieties and for an endogenous maize (invertase) gene system.

  18. Ambystoma maculatum (spotted salamander). Reproduction

    USGS Publications Warehouse

    Glorioso, Brad M.; Waddle, Hardin; Hefner, Jeromi

    2012-01-01

    The Spotted Salamander is a wide-ranging salamander of the eastern United States that typically breeds in winter or early spring in ephemeral pools in lowland forests. Ambystoma maculatum is known to deposit 2-4 egg masses per year, each containing 1-250 eggs. As part of ongoing research into the ecology and reproductive biology of Spotted Salamanders in the Kisatchie District of Kisatchie National Forest in Natchitoches Parish, Louisiana, USA, we have been counting the number of embryos per egg mass. We captured seven female A. maculatum in a small pool, six of which were still gravid. We took standard measurements, including SVL, and then implanted a Passive Integrated Transponder (PIT tag) into each adult female as was the protocol. About an hour after processing these animals we marked new A. maculatum egg masses found in the same small pool using PVC pin flags pushed carefully through the outer jelly. We did not have enough time to process them that evening, and it was not until a few days later that we photographed those masses. We discovered that one of the masses contained a PIT tag in the outer jelly that corresponded to one of the six gravid females that were marked that same evening. To our knowledge, this is the first report of PIT tags being the means, albeit coincidentally, by which a particular egg mass of Ambystoma maculatum has been assigned to a particular female. For our purposes, losing the PIT tag from the adult female is counter to the goals of our study of this population, and we will no longer be implanting PIT tags into gravid females.

  19. Development of transgenic sorghum for insect resistance against the spotted stem borer (Chilo partellus).

    PubMed

    Girijashankar, V; Sharma, H C; Sharma, Kiran K; Swathisree, V; Prasad, L Sivarama; Bhat, B V; Royer, Monique; Secundo, Blanca San; Narasu, M Lakshmi; Altosaar, I; Seetharama, N

    2005-11-01

    Transgenic sorghum plants expressing a synthetic cry1Ac gene from Bacillus thuringiensis (Bt) under the control of a wound-inducible promoter from the maize protease inhibitor gene (mpiC1) were produced via particle bombardment of shoot apices. Plants were regenerated from the transformed shoot apices via direct somatic embryogenesis with an intermittent three-step selection strategy using the herbicide Basta. Molecular characterisation based on polymerase chain reaction and Southern blot analysis revealed multiple insertions of the cry1Ac gene in five plants from three independent transformation events. Inheritance and expression of the Bt gene was confirmed in T(1) plants. Enzyme-linked immunosorbant assay indicated that Cry1Ac protein accumulated at levels of 1-8 ng per gram of fresh tissue in leaves that were mechanically wounded. Transgenic sorghum plants were evaluated for resistance against the spotted stem borer (Chilo partellus Swinhoe) in insect bioassays, which indicated partial resistance to damage by the neonate larvae of the spotted stem borer. Reduction in leaf damage 5 days after infestation was up to 60%; larval mortality was 40%, with the surviving larvae showing a 36% reduction in weight over those fed on control plants. Despite the low levels of expression of Bt delta-endotoxin under the control of the wound-inducible promoter, the transgenic plants showed partial tolerance against first instar larvae of the spotted stem borer.

  20. Biochar mitigation of allelopathy induced yield loss in continuous maize