molecular phylogenetic relationships of china seas groupers based on cytochrome b gene fragment sequences.the classification and evolutionary relationships are important issues in the study of the groupers. cytochrome b gene fragment of twenty-eight grouper species within six genera of subfamily epinephelinae was amplified using pcr techniques and the sequences were analyzed to derive the phylogenetic relationships of the groupers from the china seas. genetic information indexes, including kimura-2 parameter genetic distance and ts/tv ratios, were generated by using a variety of biology softwares. w ...200616856492
infection competition against grouper nervous necrosis virus by virus-like particles produced in escherichia coli.dragon grouper (epinephelus lanceolatus) nervous necrosis virus (dgnnv) comprises 180 copies of capsid protein that encapsulate a bipartite genome of single-stranded (+)-rnas. this study reports that virus-like particles (vlps) are formed in escherichia coli expressing the full-length orf encoding the dgnnv capsid protein. two sizes of vlps are observed. the heavier particles resemble the native piscine nodavirus in size and stain permeability, while the lighter ones are approximately two-thirds ...200312771428
involvement of the terminus of grouper betanodavirus capsid protein in virus-like particle assembly.dragon grouper, epinephelus lanceolatus, nervous necrosis virus (dgnnv) consists of 180 copies of capsid protein that encapsulates a bipartite genome of single-stranded (+)rnas. expressing the open reading frame (orf) of rna2 in escherichia coli forms virus-like particles (vlps) that resembles native virus. deleting n- and c-termini revealed different impacts on vlp formation. deletion of 35 or 52 residues at the n-terminus completely ruined the vlp assembly, presumably due to removal of positiv ...200312556997
characterization of virus-like particles assembled in a recombinant baculovirus system expressing the capsid protein of a fish nodavirus.betanodaviruses are causative agents of neurological disorders in several species of fish. we cloned and sequenced the rna2 segment of two grouper viruses isolated from epinephelus malabaricus (malabaricus grouper nervous necrosis virus, mgnnv) and epinephelus lanceolatus (dragon grouper nervous necrosis virus, dgnnv). the sequences of the two rnas were 99% identical and comparison with previously sequenced rna2 segments of fish nodaviruses striped jack nervous necrosis virus, atlantic halibut v ...200111883005
pcr amplification and sequence analysis of the major capsid protein gene of megalocytiviruses isolated in taiwan.viruses belonging to the genus megalocytivirus in the family iridoviridae are one of the major agents causing mass mortalities in marine and freshwater fish in asian countries. outbreaks of iridovirus disease have been reported among various fish species in taiwan. however, the genotypes of these iridoviruses have not yet been determined. in this study, seven megalocytivirus isolates from four fish species: king grouper, epinephelus lanceolatus (bloch), barramundi perch, lates calcarifer (bloch) ...200919460084
subset selection of high-depth next generation sequencing reads for de novo genome assembly using mapreduce framework.recent progress in next-generation sequencing technology has afforded several improvements such as ultra-high throughput at low cost, very high read quality, and substantially increased sequencing depth. state-of-the-art high-throughput sequencers, such as the illumina miseq system, can generate ~15 gbp sequencing data per run, with >80% bases above q30 and a sequencing depth of up to several 1000x for small genomes. illumina hiseq 2500 is capable of generating up to 1 tbp per run, with >80% bas ...201526678408
recombinant production of biologically active giant grouper (epinephelus lanceolatus) growth hormone from inclusion bodies of escherichia coli by fed-batch culture.growth hormone (gh) performs important roles in regulating somatic growth, reproduction, osmoregulation, metabolism and immunity in teleosts, and thus, it has attracted substantial attention in the field of aquaculture application. herein, giant grouper gh (gggh) cdna was cloned into the pet28a vector and expressed in shuffle® t7 competent escherichia coli. recombinant n-terminal 6× his-tagged gggh was produced mainly in insoluble inclusion bodies; the recombinant gggh content reached 20% of tot ...201525703054
permanent genetic resources added to molecular ecology resources database 1 august 2009-30 september 2009.this article documents the addition of 238 microsatellite marker loci and 72 pairs of single nucleotide polymorphism (snp) sequencing primers to the molecular ecology resources database. loci were developed for the following species: adelges tsugae, artemisia tridentata, astroides calycularis, azorella selago, botryllus schlosseri, botrylloides violaceus, cardiocrinum cordatum var. glehnii, campylopterus curvipennis, colocasia esculenta, cynomys ludovicianus, cynomys leucurus, cynomys gunnisoni, ...201021565018
recombinant expression of epinephelus lanceolatus serum amyloid a (elsaa) and analysis of its macrophage modulatory activities.serum amyloid a (saa) is an acute-phase protein that plays a crucial role in the inflammatory response. in this study, we identified an saa homolog from epinephelus lanceolatus (elsaa). molecular characterization revealed that elsaa contains a fibronectin-like motif that is typical of saas. recombinant elsaa protein (relsaa) was produced in e. coli bl21 (de3) cells and purified as a soluble protein. to analyze its biological activity, mouse raw264.7 macrophage cells were treated with various con ...201728323212
exophiala xenobiotica aerocystitis in a queensland grouper epinephelus lanceolatus (bloch). 201524397705
establishment of a new cell line from the heart of giant grouper, epinephelus lanceolatus (bloch), and its application in toxicology and virus susceptibility.a new marine fish cell line, derived from the heart of giant grouper, epinephelus lanceolatus (bloch), was established and characterized. the cell line was designated as elgh and subcultured with more than 60 passages. the elgh cells were mainly composed of fibroblast-like cells and multiplied well in leibovitz's l-15 medium supplemented with 10% foetal bovine serum (fbs) at 28 °c. chromosome analysis indicated that the modal chromosome number was 48. the fluorescent signals were detected in elg ...201524372271
crystallization and x-ray diffraction of virus-like particles from a piscine betanodavirus.dragon grouper nervous necrosis virus (dgnnv), a member of the genus betanodavirus, causes high mortality of larvae and juveniles of the grouper fish epinephelus lanceolatus. currently, there is no reported crystal structure of a fish nodavirus. the dgnnv virion capsid is derived from a single open reading frame that encodes a 338-amino-acid protein of approximately 37 kda. the capsid protein of dgnnv was expressed to form virus-like particles (vlps) in escherichia coli. the vlp shape is t = 3 q ...201425084387
immune response against grouper nervous necrosis virus by vaccination of virus-like particles.the grouper is a high-value fish in the seafood market. grouper nervous necrosis virus (gnnv) causes mass mortality, near 100% in larvae and juveniles, which has great economic impact on the aquaculture of the marine fish. since vaccination is one of the best methods against viral diseases, grouper epinephelus lanceolatus was injected with virus-like particles (vlps) of gnnv at different dosages and injection frequencies. the anti-sera of vaccinated fish were analyzed with antigen-capture elisa ...200616797803
infection and pathology in queensland grouper, epinephelus lanceolatus, (bloch), caused by exposure to streptococcus agalactiae via different routes.since 2007, 96 wild queensland groupers, epinephelus lanceolatus, (bloch), have been found dead in ne australia. in some cases, streptococcus agalactiae (group b streptococcus, gbs) was isolated. at present, a gbs isolate from a wild grouper case was employed in experimental challenge trials in hatchery-reared queensland grouper by different routes of exposure. injection resulted in rapid development of clinical signs including bilateral exophthalmia, hyperaemic skin or fins and abnormal swimmin ...201525117665
natural outbreak of streptococcus agalactiae (gbs) infection in wild giant queensland grouper, epinephelus lanceolatus (bloch), and other wild fish in northern queensland, australia.ninety-three giant queensland grouper, epinephelus lanceolatus (bloch), were found dead in queensland, australia, from 2007 to 2011. most dead fish occurred in northern queensland, with a peak of mortalities in cairns in june 2008. in 2009, sick wild fish including giant sea catfish, arius thalassinus (rüppell), and javelin grunter, pomadasys kaakan (cuvier), also occurred in cairns. in 2009 and 2010, two disease epizootics involving wild stingrays occurred at sea world marine aquarium. necropsy ...201222324342
transcriptome analysis of the effect of vibrio alginolyticus infection on the innate immunity-related tlr5-mediated induction of cytokines in epinephelus lanceolatus.epinephelus lanceolatus, considered to be an aquaculture fish species of high economic value in east asia, is one of the largest groupers in the epinephelus genus. vibrio alginolyticus is a bacterial species that causes high morbidity in marine fish; infection can cause exophthalmia, ulcers, septicemia, and corneal opaqueness in fish. epinephelus lanceolatus larvae infected with vibrio alginolyticus were subjected to transcriptome analysis to study the immune regulation pathway. grouper larvae w ...201626975410
isolation, identification and pathogenicity of vibrio harveyi, the causal agent of skin ulcer disease in juvenile hybrid groupers epinephelus fuscoguttatus × epinephelus lanceolatus.the hybrid grouper, epinephelus fuscoguttatus (♀) × epinephelus lanceolatus (♂), is a newly bred cultivated marine fish species of high economic value. however, a skin ulcer disease with high mortality has occurred, and the responsible pathogen remains unknown. in this study, we summarized the epidemic status and external signs of this disease. we screened potential pathogens and finally isolated one bacterial strain ml01 from affected fish. we subjected healthy juvenile hybrid groupers to bacte ...201728252178
sperm of the giant grouper: cryopreservation, physiological and morphological analysis and application in hybridizations with red-spotted order to develop excellent germplasm resources for giant grouper (epinephelus lanceolatus), cryopreservation of giant grouper sperm was examined in the present study. firstly, 13 kinds of sperm dilution (els1-3, em1-2, ts-2, mprs, elrs0-6) were prepared with physiological salt, sucrose, glucose and fetal bovine serum. the physiological parameters of elrs3 (ratio of fast motion, ratio of slow motion, time of fast motion, time of slow motion, lifespan and motility) and els3 (sperm ratio of slow ...201525985804
culture-independent genomics of a novel chlamydial pathogen of fish provides new insight into host-specific adaptations utilized by these intracellular bacteria.several chlamydiales families are associated with epitheliocystis, a common condition of the fish gill epithelium. these families share common ancestors with the chlamydiaceae and environmental chlamydiae. due to the lack of culture systems, little is known about the biology of these chlamydial fish pathogens. we investigated epitheliocystis in cultured orange-spotted grouper (epinephelus coioides) from north queensland, australia. basophilic inclusions were present in the gills of 22/31 fish an ...201728205377
formation of diploid and triploid hybrid groupers (hybridization of epinephelus coioides ♀ × epinephelus lanceolatus ♂) and their 5s gene analysis.interspecies hybridization is widely used to achieve heterosis or hybrid vigor, which has been observed and harnessed by breeders for centuries. natural allopolyploid hybrids generally exhibit more superior heterosis than both the diploid progenies and their parental species. however, polyploid formation processes have been long ignored, the genetic basis of heterosis in polyploids remains elusive.201627717311
sperm cryopreservation in different grouper subspecies and application in interspecific hybridization.the present study established and confirmed an efficient technology for groupers: giant grouper epinephelus lanceolatus, orange-spotted grouper e. coioides, seven-band grouper, e. septemfasciatus, and kelp grouper e. moara sperm cryopreservation and successfully applied the cryopreserved e. lanceolatus sperm into interspecific hybridization with e. coioides. for both e. lanceolatus and e. coioides, the best motility of postthaw sperm were achieved using 6% to 10% dmso, 6% to 16% propylene glycol ...201626803184
characterization of alpha-2-macroglobulin from groupers.alpha-2-macroglobulin (α-2-m) is a protease inhibitor broadly present in the plasma of vertebrates and invertebrates, and is an important non-specific humoral factor in defence system of the animals. this study conducted the immuno-analysis and mass spectrometric analysis methods to investigate the characteristics of the protease inhibitor, α-2-m, among groupers and related species. rabbit antiserum to the purified α-2-m of epinephelus coioides was used in different immunological methods to dete ...201323711467
complete mitochondrial genome of yellow grouper epinephelus awoara (perciformes, epinephelidae).in this paper, the complete mitochondrial (mt) genome of epinephelus awoara has been determined. the mt genome (16,802 bp) had the canonical mt gene content and arrangement, including 13 protein-coding genes, 2 rrna genes, 22 trna genes, and 2 non-coding regions. the overall base composition of the heavy-strand is 28.47% a, 27.29% t, 16.54% g, and 27.70% c, with an at content of 56.76%. it shared 92.6%, 85.5%, 86.1%, 84.9%, and 85.7% identities with that of epinephelus akaara, epinephelus bruneu ...201222943451
[microsatellite analysis of genetic diversity and phylogenetic relationship of nine species of grouper in genus epinephelus].thirteen microsatellite markers of epinephelus awoara previously discovered by our lab were selected to analyze the genetic diversity and phylogenetic relationship of nine species of grouper (e. awoara, e. merra, e. fario, e. fasciatus, e. lanceolatus, e. akaara, e. septemfasciatus, e. coioides and e. fuscoguttatus) from south china sea. the results showed that the number of total alleles of these 13 microsatellite loci was 84 in these fishes, the mean number of alleles ranged from 2.69 to 5.38, ...200717646150
growth hormone and two forms of insulin-like growth factors i in the giant grouper (epinephelus lanceolatus): molecular cloning and characterization of tissue distribution.growth hormone and insulin-like growth factors play important roles in the growth, development and metabolism of vertebrates. in this study, we used reverse transcription and rapid amplification of cdna ends (race) to obtain the three full-length cdna sequences encoding gh and two forms of igf-i from the giant grouper (epinephelus lanceolatus), a coral fish of high commercial value cultured in southeast asia. gh precursor cdna consists of 938 bp in size with an open-reading frame (orf) encoding ...201020467861
roe protein hydrolysates of giant grouper (epinephelus lanceolatus) inhibit cell proliferation of oral cancer cells involving apoptosis and oxidative stress.roe protein hydrolysates were reported to have antioxidant property but the anticancer effects were less addressed, especially for oral cancer. in this study, we firstly used the ultrafiltrated roe hydrolysates (urh) derived from giant grouper (epinephelus lanceolatus) to evaluate the impact of urh on proliferation against oral cancer cells. we found that urh dose-responsively reduced cell viability of two oral cancer cells (ca9-22 and cal 27) in terms of atp assay. using flow cytometry, urh-ind ...201627195297
patterns of coral-reef finfish species disappearances inferred from fishers' knowledge in global epicentre of marine shorefish the philippines, very high fishing pressure coincides with the globally greatest number of shorefish species, yet no long-term fisheries data are available to explore species-level changes that may have occurred widely in the most species rich and vulnerable marine ecosystem, namely coral reefs. through 2655 face-to-face interviews conducted between august 2012 and july 2014, we used fishers' recall of past catch rates of reef-associated finfish to infer species disappearances from catches in ...201627191602
development of novel tetra- and trinucleotide microsatellite markers for giant grouper epinephelus lanceolatus using 454 pyrosequencing.giant grouper (epinephelus lanceolatus) is a commercially important species, but its wild population has recently been classified as vulnerable. this species has significant potential for use in aquaculture, though a greater understanding of population genetics is necessary for selective breeding programs to minimize kinship for genetically healthy individuals. high-throughput pyrosequencing of genomic dna was used to identify and characterize novel tetra- and trinucleotide microsatellite marker ...201627059503
the complete mitochondrial genome of the epinephelus lanceolatus (perciformes: serranidae).the complete mitochondrial genome of the epinephelus lanceolatus was presented in this study. the mitochondrial genome is 16,743 bp long and consists of 13 protein-coding genes, two rrna genes, 22 trna genes and a control region. the gene order and composition of e. lanceolatus mitochondrial genome was similar to that of most other vertebrates. the nucleotide compositions of the light strand are 26.55% of a, 15.02% of c, 29.67% of t and 28.76% of g. with the exception of the nadh dehydrogenase s ...201625286235
serranus magnificus macleay 1882, a junior synonym of epinephelus lanceolatus (bloch 1790) (teleostei: serranidae). 201425283126
effect of codon optimisation on the production of recombinant fish growth hormone in pichia pastoris.this study was established to test the hypothesis of whether the codon optimization of fish growth hormone gene (fgh) based on p. pastoris preferred codon will improve the quantity of secreted rfgh in culture supernatant that can directly be used as fish feed supplements. the optimized fgh coding sequence (ofgh) and native sequence (nfgh) of giant grouper fish (epinephelus lanceolatus) were cloned into p. pastoris expression vector (ppiczαa) downstream of alcohol oxidase gene (aox1) for efficien ...201425147851
development of pedigree classification using microsatellite and mitochondrial markers for giant grouper broodstock (epinephelus lanceolatus) management in taiwan.most giant groupers in the market are derived from inbred stock. inbreeding can cause trait depression, compromising the animals' fitness and disease resistance, obligating farmers to apply increased amounts of drugs. in order to solve this problem, a pedigree classification method is needed. here, microsatellite and mitochondrial dna were used as genetic markers to analyze the genetic relationships among giant grouper broodstocks. the 776-bp fragment of high polymorphic mitochondrial d-loop seq ...201424796300
application maldi tof on protein identification of vitellogenin in giant grouper (epinephelus lanceolatus).a new proteomics technology has been implemented to study the protein repertoires of developing oocytes of giant grouper (epinephelus lanceolatus). knowledge of the chemical composition and physiochemical properties of vitellogenin (vtg) is necessary to interpret the functional and biological properties attributed during ovulation. vtg, as a biomarker indicator in sex determination, has been analyzed to determine the sex and maturational status of fish in the absence of the gonad tissue. a male ...201323494207
development and characterization of 32 microsatellite loci in the giant grouper epinephelus lanceolatus (serranidae).an economically important marine fish species, the giant grouper epinephelus lanceolatus (serranidae) is widely cultured in taiwan and costal areas of china. we isolated and characterized 32 polymorphic microsatellite loci from a ca-enriched genomic library of giant grouper. the number of alleles per locus ranged from 3 to 7, with a mean of 4.69. observed and expected heterozygosities per locus varied from 0.387 to 1.000 and from 0.377 to 0.843, respectively. six loci significantly deviated from ...201122194200
characterization of eight polymorphic microsatellite loci for the giant grouper (epinephelus lanceolatus bloch).eight polymorphic microsatellite loci were isolated and characterized using a small insert genomic dna library for the giant grouper (epinephelus lanceolatus bloch, 1790), a commercially valuable marine fish in tropical waters. they showed polymorphism information content ranging from 0.177 to 0.775, allele numbers ranging from two to 10, effective allele numbers ranging from 1.227 to 5.012, and observed and expected heterozygosities from 0.2 to 0.733 and from 0.185 to 0.801, respectively, which ...200821585897
susceptibility of farmed juvenile giant grouper epinephelus lanceolatus to a newly isolated grouper iridovirus (genus ranavirus).a ranavirus was isolated from the diseased farmed groupers (grouper iridovirus in genus ranavirus, giv-r), epinephelus hybrids (blotchy rock cod, epinephelus fuscoguttatus ♀×giant grouper, epinephelus lanceolatus ♂), in sanya, hainan, in july 2013. in this study, susceptibility of farmed juvenile giant grouper e. lanceolatus to giv-r was determined by intraperitoneally injection. the cumulative mortality reached to 81% at 5 day post infection. histologically, severe degeneration with massive pyc ...201525912024
decreasing salinity of seawater moderates immune response and increases survival rate of giant groupers post betanodavirus infection.giant groupers (epinephelus lanceolatus), an important aquaculture fish in asia, are attacked by nervous necrosis virus (nnv), belonging to betanodavirus. environmental salinity can affect fish immunity and physiology. we examined whether decreasing salinity from 30 to 15 ppt during acclimation of groupers could affect survival with nnv infection and the associated factors. although nnv infection decreased muscle moisture, up-regulated the gene expression of na(+)-k(+)-2cl(-) cotransporter isofo ...201627569983
microsatellite analysis of the genetic relationships between wild and cultivated giant grouper in the south china sea.the giant grouper (epinephelus lanceolatus) is a coral fish with high commercial value in southeast asia. in the present study, we isolated 11 microsatellite dna markers, and analysed the genetic diversity and differentiation between cultured stocks and wild populations of the giant grouper originating from the south china sea. a total of 390 alleles at 11 microsatellite loci were detected in 130 individuals from five different populations. the expected heterozygosity varied from 0.131 to 0.855 ...201627350681
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