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a new virus isolate from infectious hypodermal and hematopoietic necrosis virus (ihhnv)-infected penaeid shrimps.a new virus was isolated from infectious hypodermal and hematopoietic necrosis virus (ihhnv)-infected penaeid shrimps. the virus was isolated from two species of penaeid shrimps obtained from three different sources employing a previously developed cell-culture assay. electron-microscopical studies of both purified virus and infected cells showed bullet-shaped particles identifying it as a rhabdovirus.19911864905
growth of the penaeid shrimp virus infectious hypodermal and hematopoietic necrosis virus in a fish cell line.an established fish cell line, epithelioma papillosum cyprini (epc), originating from the carp (cyprinus carpio) was found to be susceptible to the penaeid shrimp virus, infectious hypodermal and hematopoietic necrosis virus (ihhnv). this economically important pathogen of penaeid shrimp raised in mariculture has been previously limited to growth only in young post-larval shrimps. this is, to our knowledge, the first reported successful growth of the penaeid virus in a fish cell line; it promise ...19902117021
purification and characterization of the infectious hypodermal and haematopoietic necrosis virus of penaeid shrimps.infectious hypodermal and haematopoietic necrosis (ihhn) is one of the most important viral diseases of cultured penaeid shrimps and is potentially a limiting factor in the development of farming projects for some species of these shrimps. although the ihhn agent was recognized early as being viral in origin, attempts to characterize it were inconclusive because of difficulties in obtaining sufficient amounts of purified virions to permit its characterization. recent improvements of purification ...19902254754
isolation, purification and characterization of infectious hypodermal and hematopoietic necrosis virus (ihhnv) from penaeidshrimp.a protocol was developed for the isolation and purification of the infectious hypodermal and hematopoietic necrosis virus (ihhnv) of penaeid shrimps. cesium chloride-banded virus obtained from three sources of infected shrimps was found to have a buoyant density of 1.33 g/cm3. also, electron microscopical studies employing negative stain revealed isometric particles having a size of 19 +/- 1 nm. colorimetric analysis of its nucleic acid type indicated that these particles contain only rna.19892621242
partial cloning of the genome of infectious hypodermal and haematopoietic necrosis virus, an unusual parvovirus pathogenic for penaeid shrimps; diagnosis of the disease using a specific probe.the infectious hypodermal and haematopoietic necrosis virus (ihhnv), pathogenic for penaeid shrimp, is an icosahedral unenveloped particle, 22 nm in diameter, with an ssdna linear genome, and proposed to be a member of the parvoviridae. a large majority of minus-strand dna is incorporated into the capsids compared to the plus-strand. a small amount of reannealed plus- and minus-strands (dsdna) obtained after nucleic acid extraction was blunt-ended and cloned into the system puc18/escherichia col ...19938277269
epizootiology, distribution and the impact on international trade of two penaeid shrimp viruses in the americas.marine penaeid shrimp are effected by approximately twenty viruses, the majority of which were discovered as a result of their negative effects on aquaculture. in the americas, infectious hypodermal and haematopoietic necrosis (ihhn) virus and taura syndrome (ts) virus have had a significant negative impact on aquaculture industries and, in one instance, on a commercial fishery. both viruses have become widely distributed as a consequence of the movement of host stocks for aquaculture. ihhn viru ...19968890382
taura syndrome of penaeid shrimp: cloning of viral genome fragments and development of specific gene probes.the ssrna genome extracted from purified taura syndrome virus (tsv) was transcribed into double-stranded, blunt-ended cdna and was used to construct cdna libraries either in puc 18 or in pbluescript ii ks-vectors. twelve recombinant plasmids chosen after screening of the libraries were subjected to restriction enzyme digestions for determination of size inserts and restriction maps. two of them, pp15 and pq1, were selected for probe construction. the inserts, 1500 and 1300 base pairs (bp) respec ...19989653455
a yellow head virus gene probe: nucleotide sequence and application for in situ hybridization.a portion of the genome of yellow head virus (yhv) of penaeid shrimp was cloned and the cdna fragment (1161 bp) was designated clone 3-27. the fragment was labeled with digoxigenin and hybridized in situ to tissue sections of yhv-infected penaeus vannamei. positively reacting tissues included those of the lymphoid organ, cuticular epithelium, and gills. in addition, connective tissue of hepatopancreas, heart, antennal gland, hematopoietic organ, nerve tract, midgut cecum and muscle reacted to th ...199910228874
a non-destructive method based on the polymerase chain reaction for detection of hepatopancreatic parvovirus (hpv) of penaeid shrimp.current methods to detect hepatopancreatic parvovirus (hpv) infection of penaeid shrimp depend on invasive techniques that require dissecting the organs infected by this virus. however, sacrificing valuable stocks in order to determine their hpv status can be a drawback in the case of breeding programs. a method was developed for hpv detection by applying a polymerase chain reaction (pcr) assay to fecal samples collected from live hpv-infected shrimp penaeus chinensis. a pair of pcr primers, 112 ...200010768285
use of polymerase chain reaction for the detection of infectious hypodermal and hematopoietic necrosis virus in penaeid shrimp.a rapid and reliable polymerase chain reaction (pcr) method was developed for the detection of infectious hypodermal and hematopoietic necrosis virus (ihhnv) in penaeid shrimp. the oligonucleotide primers amplify a 1681-bp fragment of ihhnv, which encompasses the coding sequence for one of the viral coat proteins. the pcr method detects ihhnv in hemolymph and homogenized tissue obtained from the cephalothorax or pleopods of infected shrimp. the technique was also successfully applied to tissue s ...200010960121
infectious hypodermal and hematopoietic necrosis virus of shrimp is related to mosquito brevidensoviruses.we purified and sequenced infectious hypodermal and hematopoietic necrosis virus (ihhnv), a small dna virus of shrimp, from wild penaeus stylirostris. the virion has a buoyant density of 1.45 as determined by cesium chloride gradient. analysis of 3873 nucleotides of the viral genome revealed three large open reading frames (orfs) and parts of the noncoding termini of the viral genome. the left, mid, and right orfs on the complementary (plus) strand have potential coding capacities of 666 amino a ...200011062047
detection and quantification of infectious hypodermal and hematopoietic necrosis virus in penaeid shrimp by real-time pcr.a real-time pcr method using a fluorogenic 5' nuclease assay and a pe applied biosystems geneamp 5700 sequence detector was developed to detect infectious hypodermal and hematopoietic necrosis virus (ihhnv) in penaeid shrimp. a pair of pcr primers to amplify an 81 bp dna fragment and a fluorogenic probe (taqman probe) were selected from orf1 (open reading frame 1) of the ihhnv genome. the primers and taqman probe used in this assay were shown to be specific for ihhnv and did not react with eithe ...200111324819
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.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. th ...200111474000
development and application of monoclonal antibodies for the detection of white spot syndrome virus of penaeid shrimp.monoclonal antibodies (mabs) were produced against white spot syndrome virus (wssv) of penaeid shrimp. the virus isolate used for immunization was obtained from china in 1994 and was passaged in penaeus vannamei. the 4 hybridomas selected for characterization all produced mabs that reacted with the 28 kd structural protein by western blot analysis. the mabs tested in dot-immunoblot assays were capable of detecting the virus in hemolymph samples collected from moribund shrimp during an experiment ...200111797911
rapd markers as predictors of infectious hypodermal and hematopoietic necrosis virus (ihhnv) resistance in shrimp (litopenaeus stylirostris).random amplified polymorphic dna (rapd) fingerprints of two shrimp populations (litopenaeus stylirostris) were compared to find genetic marker(s) that may be associated with infectious hypodermal and hematopoietic necrosis virus (ihhnv) resistance or susceptibility. of the 100 10-mer random primers and 100 intersimple-sequence repeat (issr) primers screened, five provided markers specific to the super shrimp population and three provided markers specific to the wild caught population. the two po ...200211908650
low sequence variation among isolates of infectious hypodermal and hematopoietic necrosis virus (ihhnv) originating from hawaii and the americas.a 2.9 kb fragment of the infectious hypodermal and hematopoietic necrosis virus (ihhnv) genome, which contains the coding sequence of putative non-structural and capsid proteins, was amplified and sequenced from each of 14 ihhnv isolates collected from cultured penaeid shrimp stocks in hawaii and various sites in the americas between 1982 and 1997. the sequence comparison indicates that the ihhnv genome is very stable, with 99.6 to 100% similarity among these 14 isolates. only nucleotide substit ...200212078987
comparison of penaeid shrimp and insect parvoviruses suggests that viral transfers may occur between two distantly related arthropod groups.the dna and putative amino acid sequences of representative insect and shrimp parvoviruses (subfamily densovirinae) were analyzed using computer programs. shrimp viruses included hepatopancreatic parvovirus (hpv) of penaeus monodon (hpvmon) and p. chinensis (hpvchin), spawner-isolated mortality virus from p. monodon (smvmon) and infectious hypodermal and hematopoietic necrosis virus (ihhnv) from p. vannamei. insect viruses included aedes aegypti densovirus (aaednv), aedes albopictus densovirus ( ...200212135792
white spot syndrome virus and infectious hypodermal and hematopoietic necrosis virus simultaneous diagnosis by miniarray system with colorimetry detection.highly sensitive and specific diagnostic tools are essential for monitoring the health status of farmed species. after the development of genomic probe diagnostic systems in the 1990s, followed by pcr-based systems, a miniarray method has been developed allowing one-step multiple detection. the miniarray method was developed to enable the accessibility of powerful array technology. to use this system, hybridisation and washing process were modified, resulting into a significant increase in the t ...200212270652
geographic variations among infectious hypodermal and hematopoietic necrosis virus (ihhnv) isolates and characteristics of their infection.nucleotide sequence variations of a 2.9 kb fragment of infectious hypodermal and hematopoietic necrosis virus (ihhnv) isolated from samples of penaeus monodon were determined and compared with an isolate from hawaii. the infection characteristics of these isolates were examined by histology, in situ hybridization, and laboratory challenge studies with p. vannamei. isolates of ihhnv were obtained from samples collected from the se asia region (the philippines, thailand, and taiwan). isolates of p ...200312650241
genetic markers applied in regression tree prediction models.classification and regression tree (cart) modelling was used to determine infectious hypodermal and haematopoietic necrosis virus (ihhnv) resistance and susceptibility in penaeus stylirostris. in a previous study, eight random amplified polymorphic dna (rapd) markers and viral load values using real-time quantitative pcr were obtained and used as the training data set in order to create numerous regression tree models. specifically, the genetic markers were used as categorical predictor variable ...200414731230
appendage deformity syndrome--a nutritional disease of macrobrachium rosenbergii.culture of the freshwater prawn macrobrachium rosenbergii as an alternative to penaeid shrimp has recently increased in coastal areas of southern india in order to avoid numerous problems, particularly with white spot syndrome virus (wssv). however, m. rosenbergii culture is now threatened by a new disease, appendage deformity syndrome (ads), that also results in high mortality. analysis of ads prawns for viruses such as wssv, monodon baculovirus (mbv) and infectious hypodermal and hematopoeitic ...200415212295
multiple pathogens found in growth-retarded black tiger shrimp penaeus monodon cultivated in thailand.in 2001-2002 throughout thailand, black tiger shrimp penaeus monodon farmers reported very unusual retarded growth. we have called this problem monodon slow growth syndrome (msgs). based on decreased national production, estimated losses due to this phenomenon were in the range of 13 000 million baht (approximately 300 million us dollars) in 2002. since rearing practices had not changed, it was considered possible that the msgs problem may have arisen from a new or existing pathogen. to examine ...200415460852
rediscovery of the australian strain of infectious hypodermal and haematopoietic necrosis virus.in this study we rediscovered and verified the presence of an infectious hypodermal and haematopoietic necrosis virus (ihhnv) strain amongst cultured penaeid prawns penaeus monodon from 1993 in australia on the basis of a pcr analysis with ihhnv specific primers and sequencing of the resulting amplicons. a total of 7 previously published diagnostic primers specific to ihhnv were tested against australian penaeid prawns and only 1 elicited a positive ihhnv pcr result with 16 out of 20 australian ...200415584422
sensitive and rapid detection of infectious hypodermal and hematopoietic necrosis virus (ihhnv) in shrimps by loop-mediated isothermal amplification.a method for nucleic acid amplification, loop-mediated isothermal amplification (lamp) is a novel, sensitive and rapid technique, which can be applied for disease diagnosis in aquaculture. using the lamp method, a highly specific and sensitive diagnostic system for infectious hypodermal and hematopoietic necrosis virus (ihhnv) detection was designed. a set of four primers was designed by targeting the ihhnv genome dna. by the detection system, target dna was amplified and visualized on agarose g ...200616214229
complete nucleotide sequence and genomic organization of hepatopancreatic parvovirus (hpv) of penaeus monodon.we have determined the genome of hepatopancreatic parvovirus (hpv), a minus, single-stranded dna virus isolated from infected penaeus monodon in thailand. its genome consisted of 6321 nucleotides, representing three large open reading frames (orfs) and two non-coding termini. the left (orf1), mid (orf2), and right (orf3) orfs on the complementary (plus) strand may code for 428, 579, and 818 amino acids, equivalent to 50, 68, and 92 kda, respectively. the 5' and 3' ends of viral genome contained ...200616356523
different responses to infectious hypodermal and hematopoietic necrosis virus (ihhnv) in penaeus monodon and p. vannamei.infectious hypodermal and hematopoietic necrosis virus (ihhnv) is widespread in cultured penaeus monodon and p. vannamei in thailand. it causes runt-deformity syndrome that is characterized by physical abnormalities and stunted growth in p. vannamei, but causes no apparent disease in p. monodon. in both species, the virus may produce cowdry type a inclusions in tissues of ectodermal and mesodermal origin, but these are common in p. vannamei and rare in p. monodon. the virus can be more easily de ...200516408834
infectious hypodermal and hematopoietic necrosis virus (ihhnv)-related sequences in the genome of the black tiger prawn penaeus monodon from africa and australia.we found an infectious hypodermal and hematopoietic necrosis virus (ihhnv)-related sequence within the shrimp genome in populations of penaeus monodon from africa and australia. ihhnv is a single-stranded dna virus that has caused severe mortality and stunted growth in penaeid shrimp. recently, ihhnv-related sequences were found in samples of p. monodon from madagascar and tanzania. these sequences vary considerably (14 and 8%, respectively) from that of ihhnv found in association with viral epi ...200616473428
development of real-time polymerase chain reaction assay with taqman probe for the quantitative detection of infectious hypodermal and hematopoietic necrosis virus from shrimp.an assay was developed for the detection of infectious hypodermal and hematopoietic necrosis virus (ihhnv) based on real-time quantitative polymerase chain reaction (pcr). a pair of primers and a taqman probe were designed that are specific for the recognition of a conservative region in the ihhnv genome. the ihhnv real-time pcr assay had a detection limit of 9 dna copies, with a dynamic range of detection between 9 x 106 and 9 dna copies. the primer pairs and probe were specific to ihhnv and di ...200616512254
a single-step multiplex pcr for simultaneous detection of white spot syndrome virus and infectious hypodermal and haematopoietic necrosis virus in penaeid shrimp.white spot syndrome virus (wssv) and infectious hypodermal and haematopoietic necrosis virus (ihhnv) are the major viral pathogens of penaeid shrimp worldwide (lightner & redman 1998). litopenaeus vannamei was introduced into china from the americas, and quickly became widely cultured. following its introduction, both ihhnv and wssv have become important pathogens of cultured penaeid shrimp and have had a huge impact on the culture industry in china in recent years.200616677320
low impact of infectious hypodermal and hematopoietic necrosis virus (ihhnv) on growth and reproductive performance of penaeus monodon.no controlled studies on the effect of infectous hypodermal and necrosis virus (ihhnv) on penaeus monodon have been previously reported. here we describe domesticated p. monodon that became positive for ihhnv and other viruses at variable levels of prevalence during cultivation in 16 open-air, earthen ponds. these were stocked with domesticated postlarvae (pl) that tested negative for 7 shrimp viruses including ihhnv at 6% prevalence in 3 checks using polymerase chain reaction (pcr) methods. the ...200616724556
white-spot syndrome virus (wssv) introduction into the gulf of mexico and texas freshwater systems through imported, frozen bait-shrimp.we analysed 20 boxes of, frozen imported bait-shrimp (china: parapenaeopsis sp. and metapenaeopsis sp.) and 8 boxes of native, frozen bait-shrimp (gulf of mexico: litopenaeus setiferus and farfantepenaeus duorarum) by rt-pcr or pcr for taura syndrome virus (tsv), yellowhead virus/gill-associated virus (yhv/gav), white-spot syndrome virus (wssv) and infectious hypodermal and hematopoietic necrosis virus (ihhnv). all 28 boxes of shrimp were negative for tsv, yhv/gav and ihhnv; 2 boxes of imported ...200616956056
pre-exposure to infectious hypodermal and haematopoietic necrosis virus or to inactivated white spot syndrome virus (wssv) confers protection against wssv in penaeus vannamei (boone) post-larvae.larvae and post-larvae of penaeus vannamei (boone) were submitted to primary challenge with infectious hypodermal and haematopoietic necrosis virus (ihhnv) or formalin-inactivated white spot syndrome virus (wssv). survival rate and viral load were evaluated after secondary per os challenge with wssv at post-larval stage 45 (pl45). only shrimp treated with inactivated wssv at pl35 or with ihhnv infection at nauplius 5, zoea 1 and pl22 were alive (4.7% and 4%, respectively) at 10 days post-infecti ...200617026668
evidence of existence of infectious hypodermal and hematopoietic necrosis virus in penaeid shrimp cultured in china.infectious hypodermal and hematopoietic necrosis virus is the causative agent of a shrimp disease which causes economic losses on a global scale. a pair of primers, i2814f/i3516r, was designed from the ihhnv genomic sequence (genbank) that encodes for structural protein corresponding to nucleotides 2814-3516, which amplifies a 703 base pair (bp) region from the virus genome. pcr amplification with the primers generated a product of the expected size from the purified ihhnv dna of litopenaeus van ...200717118584
viral interference between infectious hypodermal and hematopoietic necrosis virus and white spot syndrome virus in litopenaeus vannamei.white spot syndrome virus (wssv) is highly virulent and has caused significant production losses to the shrimp culture industry over the last decade. infectious hypodermal and hematopoietic necrosis virus (ihhnv) also infects penaeid shrimp and, while being less important than wssv, remains a major cause of significant production losses in litopenaeus vannamei (also called penaeus vannamei) and l. stylirostris (also called penaeus stylirostris). these 2 viruses and their interactions were previo ...200617140141
detection of infectious myonecrosis virus (imnv) of penaeid shrimp by reverse-transcriptase polymerase chain reaction (rt-pcr).infectious myonecrosis virus (imnv) infecting cultured litopenaeus vannamei in brazil is a double-stranded rna virus that causes a slowly progressive disease with cumulative mortalities of up to 70%. the disease is currently diagnosed using a combination of gross signs (primarily skeletal tail muscle necrosis with white opaque discoloration), histopathology, and in situ hybridization with a digoxigenin-labeled gene probe. a rapid and sensitive method for definitive diagnosis of the disease was d ...200617240754
detection of infectious hypodermal and hematopoietic necrosis virus (ihhnv) in litopenaeus vannamei by ramification amplification assay.infectious hypodermal and hematopoietic necrosis virus (ihhnv) is a single-stranded dna virus that causes developmental and growth abnormalities in pacific white shrimp litopenaeus vannamei (also known as penaeus vannamei). nucleic acid based methods such as in situ hybridization (ish) and pcr have been commonly used for ihhnv detection. ramification amplification (ram), an isothermal nucleic acid amplification approach, was used in this study to detect ihhnv in l. vannamei. ram offers many adva ...200617260829
pcr assay for discriminating between infectious hypodermal and hematopoietic necrosis virus (ihhnv) and virus-related sequences in the genome of penaeus monodon.we developed a pcr assay that can detect infectious hypodermal and hematopoietic necrosis virus (ihhnv) but that does not react with ihhnv-related sequences in the genome of penaeus monodon from africa and australia. ihhnv is a single-stranded dna virus that has caused severe mortality and stunted growth in penaeid shrimp. recently, ihhnv-related sequences were found in the genome of some stocks of p. monodon from africa and australia. these virus-related sequences have a high degree of similari ...200717432046
functional characterization of putative promoter elements from infectious hypodermal and hematopoietic necrosis virus (ihhnv) in shrimp and in insect and fish cell lines.infectious hypodermal and hematopoietic necrosis virus (ihhnv) of shrimp contains a linear single-stranded dna genome of approximately 4.1kb with three putative open reading frames (orfs) on the same dna strand designated, the left, middle, and right orfs. the left orf codes for non-structural protein and the right orf codes for capsid protein, whereas the role of the middle orf is still unknown. two putative promoters, designated p2 and p61, were detected upstream of the left orf and right orf, ...200717434223
a multiplex rt-pcr for simultaneous differentiation of three viral pathogens of penaeid shrimp.a multiplex reverse transcription polymerase chain reaction (mrt-pcr) was developed and optimized to simultaneously detect 3 viral pathogens of shrimp. three sets of specific oligonucleotide primers for taura syndrome virus (tsv), white spot syndrome virus (wssv) and infectious hypodermal and hematopoietic necrosis virus (ihhnv) were used in the assay. the mrt-pcr dna products were visualized by gel electrophoresis and consisted of fragments of 231 bp for tsv, 593 bp for wssv and 356 bp for ihhn ...200717718168
multiplex rt-pcr assay for simultaneous detection of six viruses of penaeid shrimp.in the present study, multiplex reverse transcription-polymerase chain reaction (mrt-pcr) was developed for simultaneously detection of six major shrimp viruses including yellow-head virus (yhv), white spot syndrome virus (wssv), taura syndrome virus (tsv), hepatopancreatic parvovirus (hpv), infectious hypodermal and hematopoietic necrosis virus (ihhnv) and monodon baculovirus (mbv). the six primer sets could amplify viral nucleic acids resulting in pcr products with different sizes. they were h ...200818406570
shrimp hepatopancreatic parvovirus detection by combining loop-mediated isothermal amplification with a lateral flow dipstick.present methods such as traditional pcr, pcr-elisa, real-time pcr and histopathology for detection of shrimp hepatopancreatic parvovirus (pmdnv) entail various disadvantages including high cost, long assay time or use of toxic substances. loop-mediated isothermal amplification (lamp) of target nucleotide sequences under inexpensive isothermal conditions combined with amplicon detection by chromatographic lateral flow dipsticks (lfd) allowed simpler detection within 75 min. biotinylated lamp ampl ...200818835299
immune response of white shrimp, litopenaeus vannamei, after a concurrent infection with white spot syndrome virus and infectious hypodermal and hematopoietic necrosis virus.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 co ...200918948207
development of a real-time multiplex pcr assay for detection of viral pathogens of penaeid shrimp.a real-time multiplex polymerase chain reaction (rtm-pcr) assay was developed and optimized to simultaneously detect three viral pathogens of shrimp in one reaction. three sets of specific oligonucleotide primers for white spot syndrome virus (wssv), infectious hypodermal and haematopoietic necrosis virus (ihhnv) and taura syndrome virus (tsv), along with three taqman probes specific for each virus were used in the assay. the rtm-pcr results were detected and analyzed using the light cycler 2.0 ...200819018451
real-time quantitative loop-mediated isothermal amplification as a simple method for detecting white spot syndrome virus.white spot syndrome virus (wssv) continues to be the most pathogenic virus among the crustacean aquaculture causing mass mortality. in the present study, we established a one-step, single tube, real-time accelerated loop-mediated isothermal amplification (real-time lamp) for quantitative detection of wssv.200919018969
detection of shrimp infectious myonecrosis virus by reverse transcription loop-mediated isothermal amplification combined with a lateral flow dipstick.infectious myonecrosis virus (imnv) has caused a slowly progressive disease with cumulative mortalities of up to 70% or more in cultured penaeus (litopenaeus) vannamei in northeast brazil and indonesia. rapid detection of viruses by loop-mediated isothermal amplification (lamp) of genomic material with high specificity and sensitivity can be applied for diagnosis, monitoring and control of diseases in shrimp aquaculture. using an imnv template, successful detection was achieved after a 60-min rt ...200919022295
expression and self-assembly of virus-like particles of infectious hypodermal and hematopoietic necrosis virus in escherichia coli.infectious hypodermal and hematopoietic necrosis virus (ihhnv) is one of the highly pathogenic shrimp viruses. in this study, ihhnv capsid protein (cp) was recombinantly expressed in escherichia coli and found to self-assemble into virus-like particles (vlps) with homogeneous size and shape similar to native ihhnv particles. furthermore, we found that ihhnv-vlps encapsidate rna and dna with a predominant size of 0.5 kb. stability experiments revealed that the vlp could not be disassembled by edt ...200919252812
molecular detection of three shrimp viruses and genetic variation of white spot syndrome virus in hainan province, china, in 2007.abstract white spot syndrome virus (wssv), taura syndrome virus (tsv) and infectious hypodermal and haematopoietic necrosis virus (ihhnv) have been responsible for major pandemics affecting the shrimp farming industry. shrimp samples were collected from eight farms in hainan province, china, during 2007 and analysed by polymerase chain reaction (pcr) or reverse transcriptase pcr methods to determine the prevalence of these viruses. from the eight sampling locations, only samples from one farm di ...200919490390
development of a real-time pcr assay for detection of monodon baculovirus (mbv) in penaeid shrimp.a real-time pcr method was developed to detect monodon baculovirus (mbv) in penaeid shrimp. a pair of mbv primers to amplify a 135bp dna fragment and a taqman probe were developed. the primers and taqman probe were specific for mbv and did not cross react with hepatopancreatic parvovirus (hpv), white spot syndrome virus (wssv), infectious hypodermal and haematopoietic virus (ihhnv) and specific pathogen free (spf) shrimp dna. a plasmid (pmbv) containing the target mbv sequence was constructed an ...200919622362
simple immunoblot and immunohistochemical detection of penaeus stylirostris densovirus using monoclonal antibodies to viral capsid protein expressed heterologously.penaeus stylirostris densovirus (pstdnv), called formerly infectious hypodermal and hematopoietic necrosis virus (ihhnv), is an important shrimp pathogen which can cause mortality in the blue shrimp penaeus (litopenaeus) stylirostris and stunting in the whiteleg shrimp penaeus (litopenaeus) vannamei. five monoclonal antibodies (mabs) were produced against the 37kda capsid protein 3 (cp3) of pstdnv expressed heterologously in the form of a fusion protein with glutathione-s-transferase called gst- ...200919654023
rapid and sensitive detection of penaeus monodon nucleopolyhedrovirus by loop-mediated isothermal amplification.loop-mediated isothermal amplification (lamp) is a novel, sensitive and rapid method for amplification of nucleic acids under isothermal conditions. in this report, a lamp method was developed for detection of penaeus monodon nucleopolyhedrovirus (pemonpv), known previously as monodon baculovirus (mbv), using a set of six primers designed to specifically recognize the pemonpv polyhedrin gene. the optimized time and temperature conditions for the lamp assay were 60 min at 63 degrees c. the sensit ...200919703492
rapid and sensitive detection of penaeus monodon nucleopolyhedrovirus (pemonpv) by loop-mediated isothermal amplification combined with a lateral-flow dipstick.several methods such as traditional pcr or nested-pcr, immuno assay and histopathology have been developed for detection of penaeus monodon nucleopolyhedrovirus (pemonpv) formerly called monodon baculovirus (mbv). however, these methods have various disadvantages including low sensitivity, long assay time, use of toxic substances or unsuitability for field diagnosis. loop-mediated isothermal amplification of target nucleotide sequences under isothermal conditions, combined with amplicon detectio ...201019818396
fast short-fragment pcr for rapid and sensitive detection of shrimp viruses.this article describes a fast short-fragment pcr method for the detection of white spot syndrome virus (wssv), infectious hypodermal and hematopoietic necrosis virus (ihhnv), and monodon baculovirus (mbv). fast two-temperature (95 degrees c denaturation and 60 degrees c annealing/extension) pcrs were performed in 5-10 microl volume samples in miniaturized microplates using a fast peltier thermal cycler. 40 cycles were completed in 25-30 min. rapid high-resolution agarose gel electrophoresis of 7 ...201020471423
attempts at producing a hybridised penaeus mondon cell line by cellular fusion.the lack of a standardised system for the isolation, identification and purification of prawn viruses, is a major obstacle to the control of viruses in penaeid aquaculture. to date, spontaneous and induced transformation of somatic penaeid cells has failed. hybrid cells with the aim of supporting the growth of penaeid viruses were created using polyethylene glycol (peg)-mediated fusion with two immortal cell lines, epithelioma papulosum cyprinid (epc) and spodoptera frugiperda pupal ovarian cell ...201020580832
genetic signature of rapid ihhnv (infectious hypodermal and hematopoietic necrosis virus) expansion in wild penaeus shrimp populations.infectious hypodermal and hematopoietic necrosis virus (ihhnv) is a widely distributed single-stranded dna parvovirus that has been responsible for major losses in wild and farmed penaeid shrimp populations on the northwestern pacific coast of mexico since the early 1990's. ihhnv has been considered a slow-evolving, stable virus because shrimp populations in this region have recovered to pre-epizootic levels, and limited nucleotide variation has been found in a small number of ihhnv isolates stu ...201020668694
[bioinformatics analysis of mosquito densovirus nostructure protein ns1].to analyze and predict the structure and function of mosquito densovirus (mdv) nostructual protein1 (ns1).200920718357
rapid and sensitive detection of infectious hypodermal and hematopoietic necrosis virus by loop-mediated isothermal amplification combined with a lateral flow dipstick.infectious hypodermal and hematopoietic necrosis virus (ihhnv) is an important shrimp pathogen that causes mortality in penaeus stylirostris and stunting (called runt deformity syndrome or rds) in penaeus vannamei. loop-mediated isothermal amplification (lamp) allows rapid amplification of nucleic acids under isothermal conditions. it can be combined with a chromatographic lateral flow dipstick (lfd) for highly specific, rapid and simple visual detection of ihhnv-specific amplicons. using this p ...201020887752
differential diagnosis of active hypodermal and hematopoietic necrosis virus based on gene choice and reverse transcription coupled with pcr.the pacific whiteleg shrimp litopenaeus vannamei (penaeidae) is one of the most important cultivated species in world aquaculture. in brazil, the northeastern states are home to the main shrimp producers. as shrimp aquaculture has expanded and intensified, diseases have progressively become one of the most serious threats to this industry. infectious hypodermal and hematopoietic necrosis virus (ihhnv) is an enzootic viral agent in brazilian shrimp farms. its is usually diagnosed by histological ...201020957606
real-time target-specific detection of loop-mediated isothermal amplification for white spot syndrome virus using fluorescence energy transfer-based probes.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. ...201121256868
prevalence of three shrimp viruses in zhejiang province in 2008.white spot syndrome virus (wssv), taura syndrome virus (tsv) and infectious hypodermal and haematopoietic necrosis virus (ihhnv) are three shrimp viruses responsible for major pandemics affecting the shrimp farming industry. shrimps samples were collected from 12 farms in zhejiang province, china, in 2008 and analyzed by pcr to determine the prevalence of these viruses. from the 12 sampling locations, 8 farms were positive for wssv, 8 for ihhnv and 6 for both wssv and ihhnv. an average percentag ...201121331893
[development of rapid detection of infectious hypodermal and hematopoietic necrosis virus by loop-mediated isothermal amplification].loop-mediated isothermal amplification (lamp) assay is a novel method of gene amplification with high specificity, sensitivity and rapidity, which can be applied for disease diagnosis in shrimp aquaculture. the method is performed under isothermal conditions with a set of four specially designed primers that recognize six distinct sequences of the target. in the present study, according to the conservative regions of non-structural protein gene ns1, a set of four specific primers were designed, ...201021344755
complete nucleic acid sequence of penaeus stylirostris densovirus (pstdnv) from india.infectious hypodermal and hematopoietic necrosis virus (ihhnv) of shrimp, recently been classified as penaeus stylirostris densovirus (pstdnv). the complete nucleic acid sequence of pstdnv from india was obtained by cloning and sequencing of different dna fragment of the virus. the genome organisation of pstdnv revealed that there were three major coding domains: a left orf (ns1) of 2001 bp, a mid orf (ns2) of 1092 bp and a right orf (vp) of 990 bp. the complete genome and amino acid sequences o ...201121402111
gene profiling and characterization of arginine kinase-1 (mrak-1) from freshwater giant prawn (macrobrachium rosenbergii).arginine kinase-1 (mrak-1) was sequenced from the freshwater prawn macrobrachium rosenbergii using illumina solexa genome analyzer technique. mrak-1 consisted of 1068 bp nucleotide encoded 355 polypeptide with an estimated molecular mass of 40 kda. mrak-1 sequence contains a potential atp:guanido phosphotransferases active domain site. the deduced amino acid sequence of mrak-1 was compared with other 7 homologous arginine kinase (ak) and showed the highest identity (96%) with ak-1 from cherry sh ...201121549198
screening of parasitic and ihhnv infections in wild giant freshwater prawn macrobrachium rosenbergii from rejang river at kuching, sarawak.a preliminary survey of parasitic and infectious hypodermal and haematopoietic necrosis virus (ihhnv) infections in giant freshwater prawn from the damak sea of rejang river, kuching, sarawak was conducted. symptoms of black spots/patches on the rostrum, carapace, pleopods or telson were observed in most of the 107 samples collected. parasitic examination revealed sessiline peritrichs such as (zoothamnium sp.), nematode larvae, gregarine stage and cocoon of leech with prevalences of 1.2%, 1.2%, ...201121602773
prophenoloxidase activating enzyme-iii from giant freshwater prawn macrobrachium rosenbergii: characterization, expression and specific enzyme activity.the prophenoloxidase activating system is an important innate immune response against microbial infections in invertebrates. the major enzyme, phenoloxidase, is synthesized as an inactive precursor and its activation to an active enzyme is mediated by a cascade of clip domain serine proteinases. in this study, a cdna encoding a prophenoloxidase activating enzyme-iii from the giant freshwater prawn macrobrachium rosenbergii, designated as mrproae-iii, was identified and characterized. the full-le ...201121614523
detection and genetic profiling of infectious hypodermal and haematopoietic necrosis virus (ihhnv) infections in wild berried freshwater prawn, macrobrachium rosenbergii collected for hatchery production.infectious hypodermal and haematopoietic necrosis virus (ihhnv) has been detected widely in penaeid culture facilities in asia and the americas. ihhnv infection on sub-adult and postlarvae of the giant freshwater prawn, macrobrachium rosenbergii which had caused up to 80% mortalities was first reported in southeast taiwan in 2006. in malaysia, although, there has been no report on ihhnv infections in m. rosenbergii, preliminary work suggests that there is an urgent need to setup a screening prot ...201121755294
additional random, single to multiple genome fragments of penaeus stylirostris densovirus in the giant tiger shrimp genome have implications for viral disease diagnosis.scattered reports of viral inserts in shrimp and insect genomes led to the hypothesis that random, autonomous insertion of such sequences occurs in these organisms and leads to specific, heritable immunity. to test the prediction regarding random insertion of viral sequences into the shrimp genome, we examined the giant tiger shrimp for random genomic insertions of penaeus stylirostris densovirus (also called ihhnv). by pcr analysis using a set of 7 overlapping primer pairs to cover the whole ih ...201121723348
bioinformatic characterization and gene expression pattern of apoptosis inhibitor from macrobrachium rosenbergii challenged with infectious hypodermal and hematopoietic necrosis virus.apoptosis is genetically programmed cellular killing processes that execute unnecessary or infected cells. it plays an important role in embryogenesis, homeostasis, insect metamorphosis and immunity. apoptosis inhibitor (mriap) was sequenced from the freshwater giant prawn macrobrachium rosenbergii using illumina solexa genome analyzer technique. mriap consisted of 1753 base pair nucleotides encoded 535 polypeptide with an estimated molecular mass of 60 kda. mriap amino acid sequence contains ia ...201121945707
genomic sequence of infectious hypodermal and hematopoietic necrosis virus (ihhnv) klv-2010-01 originating from the first korean outbreak in cultured litopenaeus vannamei.due to the need to track and monitor genetic diversity, the genome of the infectious hypodermal and hematopoietic necrosis virus (ihhnv) strain klv-2010-01 in cultured litopenaeus vannamei shrimp that originated from the first korean outbreak in 2010 was sequenced and analyzed. the genome, with a length of 3914 nucleotides, was sequenced from the korean ihhnv. the genome encoded three large and overlapping open reading frames: orf1 (ns-1) of 2001 bp, orf2 (ns-2) of 1092 bp and orf3 (capsid prote ...201122038073
effect of infectious hypodermal and haematopoietic necrosis virus (ihhnv) infection on caspase 3c expression and activity in freshwater prawn macrobrachium rosenbergii.caspase 3c (mrcasp3c) was sequenced from the freshwater giant prawn macrobrachium rosenbergii using illumina solexa genome analyzer technique. mrcasp3c consisted of 2080 bp nucleotide encoded 521 polypeptide with an estimated molecular mass of 59 kda. mrcasp3c sequence contains caspase family p20 domain profile and caspase family p10 domain profile at 236-367 and 378-468 respectively. the quantitative real time pcr analysis revealed a broad expression of mrcasp3c with the highest expression in h ...201222119573
survey of protozoan, helminth and viral infections in shrimp litopenaeus setiferus and prawn macrobrachium acanthurus native to the jamapa river region, mexico.we surveyed protozoan and metazoan parasites as well as white spot syndrome virus (wssv) and infectious hypodermal hematopoietic necrosis virus (ihhnv) in white shrimp litopenaeus setiferus and the palaemonid prawn macrobrachium acanthurus native to the lower jamapa river region of veracruz, mexico. the presence of parasites and the infection parameters were evaluated in 113 palaemonid prawns collected during the northwind (n = 45), rainy (n = 38) and dry seasons (n = 30) between october 2007 an ...201122013749
the infectious hypodermal and haematopoietic necrosis virus: a brief review of what we do and do not know.given its high prevalence, its wide distribution and its remarkable capacity to cause severe mortality in shrimp, the infectious hypodermal and haematopoietic necrosis virus (ihhnv) may deserve far more attention than it has received, as it remains considered as one of the most serious problems plaguing the global shrimp farming industry. furthermore, its real measurable impact over wild shrimp populations remains unknown. undeniably, the progress that we have reached today on the knowledge of i ...201122390574
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