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Development and application of quantitative detection method for nervous necrosis virus(NNV) isolated from sevenband grouper Hyporthodus septemfasciatus

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Asian Pacific Journal of Tropical Medicine 2016年第8期9卷 720-726页

作者： Jong-Oh Kim Jae-Ok Kim Wi-Sik Kim Myung-Joo Oh Department of Aqualife Medicine College of Fisheries and Ocean Science Chonnam National University

Objective:To develop the rapid and efficient quantitative detection tool for nervous necrosis virus isolated from sevenband grouper Hyporhodus ***:The viral genes of the NNV(SGYeosu08) isolated from sevenband grouper were phylogenetically *** addition,novel quantitative PCR primers based on the genomic sequence of SGYeosu08 isolate were designed and compared it with the conventional bio-assay method(TCID_(50)) using in vitro and in vivo ***:The phylogenetic analysis of viral genes demonstrated the relationship of SGYeosu08 with members of red-spotted grouper nervous necrosis virus(RGNNV).The qNNV_Rl primer set(R1_F and R1_R) and the qNNV_R2 primer set(R2_F and R2_R) revealed 93%primer efficiency(regression:y=-0.2861 x + 9.9401,R^2= 0.9976)and the revealed 108%primer efficiency(regression:y=-0.3172 x + 10.0611,R^2= 0.9982),*** comparison with viral infectivity calculated by TCID_(50) method showed similar kinetic pattern at in vitro and NNV challenged fish(in vivo) ***:Result show that this method is rapid and efficient to diagnose NNV infection compare to traditional bioassay method(TCID_(50)).

关键词： Nervous necrosis virus(NNV) Quantitative detection Diagnostic Sevenband grouper

Exploration of the evolution and production of volatile compounds in grouper (Epinephelus coioides) during cold storage

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Food Bioscience 2023年第2期52卷 1222-1230页

作者： Yuanming Chu Zhaoyang Ding Jinfeng Wang Jing Xie College of Food Science&Technology Shanghai Ocean UniversityShanghaiChina Shanghai Professional Technology Service Platform on Cold Chain Equipment Performance and Energy Saving Evaluation Shanghai Ocean UniversityShanghaiChina National Experimental Teaching Demonstration Center for Food Science and Engineering Shanghai Ocean UniversityShanghaiChina Shanghai Engineering Research Center of Aquatic Product Processing&Preservation Shanghai Ocean UniversityShanghaiChina Collaborative Innovation Center of Seafood Deep Processing Ministry of EducationDalian116034China

Microbial growth,protein degradation and lipid oxidation played an important role in the flavor of grouper during cold *** order to study the relationship between volatile compounds(VOCs)and microorganisms,proteins and lipids in grouper during cold storage,headspace-solid phase microextraction(HS-SPME)combined with gas chromatography-mass spectrometry(GC-MS)was used to analyze the VOCs of grouper during cold *** total viable counts(TVC),Pseudomonas count,total sulfhydryl(SH)content,carbonyl content,TBARs,total free fatty acids(FFAs)content and free amino acids(FAAs)were *** results showed that the indicators related to microorganisms,proteins and lipids deteriorated to different degrees during cold storage.A total of 67 VOCs were identified in the *** screening,10 key VOCs were found,8 of which had a negative odor and were highly correlated with the deterioration of grouper *** further correlation analysis,the source of key VOCs was basically determined,and the main factors affecting VOCs production were determined to be microbial growth,followed by protein degradation and lipid *** laid a foundation for regulating the flavor formation of grouper during cold storage from the source in the future.

关键词： grouper HS-SPME-GC-MS Volatile components Quality Cold storage

Genetic population structure of red snapper(Lutjanus malabaricus)and orange-spotted grouper(Epinephelus coiodes)in Brunei and Sabah

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Integrative Zoology 2008年第3期3卷 208-215页

作者： Zohrah H.SULAIMAN Khadijah H.ABDUL RAHMAN Teo Y.YING Hussein H.TAHA NurQistina H.MUHD TAHA Department of Biology Faculty of ScienceUniversity Brunei DarussalamJalan Tungku LinkGadongBrunei Darussalam

The genetic population structure of red snapper Lutjanus malabaricus and orange-spotted grouper Epinepheluscoiodes in Brunei and Sabah was investigated using allozyme electrophoresis. Samples were collected from threesites in Brunei for E. coiodes and from three sites in Brunei and Sabah for L. malabaricus. A total of 22 loci and 16loci were scored, respectively. The index of fixation (FST) for the E. coiodes population was 0.176 but, in general, itlacked within-population structuring. The FST was particularly high between Brunei Bay/Jerudong and Brunei Bay/Kuala Belait, suggesting genetic subdivision on a small spatial scale. Isolation of Brunei Bay from the South ChinaSea may constrain the movement of adult fishes and larval dispersal, thereby reducing homogeneity amongsubpopulations. All variable loci for E. coiodes were in Hardy–Weinberg equilibrium except for MDH* and GTDHP*(P grouper resources inBrunei waters must take into account the presence of a single stock and two stocks, respectively.

关键词： allozymes fishery resource genetic structure of fish grouper snapper

Genomic selection of orange-spotted grouper(Epinephelus coioides)based on multiplex PCR enrichment capture sequencing

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Aquaculture and Fisheries 2023年第6期8卷 681-688页

作者： Xinxin Shan Xinhui Zhang Zhiqiang Ruan Jieming Chen Qiong Shi Junmin Xu Xinxin You Shenzhen Key Lab of Marine Genomics Guangdong Provincial Key Lab of Molecular Breeding in Marine Economic AnimalsBGI Academy of Marine SciencesBGI MarineBGIShenzhen518083China Southern Marine Science and Engineering Guangdong Laboratory(Zhuhai) State Key Laboratory of BiocontrolSchool of Marine SciencesSun Yat-Sen UniversityGuangzhou510275China College of Life Sciences University of Chinese Academy of SciencesBeijing100049China

Orange spotted grouper(Epinephelus coioides)is an important mariculture fish,and genomic breeding of this grouper species has been hindered due to lack of efficient genotyping ***,we developed a single nucleotide polymorphism(SNP)genotyping technology based on multiplex PCR enrichment capture sequencing,which mainly aims at target area for high-throughput sequencing,and 741 SNPs were designed for genomic selection(GS)of growth and ammonia tolerance traits at the same *** multiplex PCR enrichment capture sequencing assay showed that the genotyping efficiency was more than 99%in the orange-spotted grouper and the predictive accuracy of body weight and ammonia tolerance traits was 82%and 96%,*** importantly,the average identity of the sequences with these SNPs aligned to the genomes of giant grouper(***)and brown-marbled grouper(***)were both over 96%.Test data showed that the SNP genotyping efficiency was more than 94%in both giant grouper and brown-marbled *** summary,these results indicated that the development of SNP loci and genotyping approach based on the multiple PCR enrichment capture sequencing are suitable for GS of growth and ammonia tolerance traits in various grouper species,and it would provide technical support for practical grouper breeding.

关键词： grouper SNP genotyping Approach Multiple PCR enrichment Capture sequencing Genomic selection

Control of gonadal maturation and sex in grouper

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Aquaculture and Fisheries 2022年第5期7卷 519-524页

作者： Yasuhisa Kobayashi Laboratory for Aquatic Biology Department of FisheriesFaculty of AgricultureKindai UniversityNakamachi 3327-204Nara631-0052Japan

Protogynous groupers,belonging to the subfamily Epinephelinae,are among the most crucial fishery targets in the Asian ***,several grouper species are cultured,providing seed for the aquaculture ***,cultivation grouper larvae are ***,the improvement of aquaculture technology of grouper production is *** managing the reproduction and sex of the target parent fish is vital to enhance the efficiency of *** paper reviews new technologies for induction of gonadal maturation and control of sex in *** dynamics of androgen-producing cells,which are essential for a sex change,are also discussed.

关键词： grouper Sex change Protogynous Gonadal maturation Androgen Estrogen

DISEASE RESISTANCE AND HUMORAL IMMUNOMODULATORY EFFECTS OF VITAMIN C ON grouper, EPINEPHELUS AWOARA

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Chinese Journal of Oceanology and Limnology 2000年第3期18卷 247-252页

作者：秦启伟吴灶和潘金培 South China Sea Institute of Oceanology Chinese Academy of Sciences Guangzhou China TMSI National University of Singapore Singapore

The disease resistance and humoral immunomodulatory effects of vitamin C administered orally to grouper, Epinephelus awoara maintained on a frozen fish diet supplemented with vitamin C at 500, 1000, 1500 and 2000 mg/kg were investigated. After 20 weeks, the growth rates of the groups with high level of vitamin C apparently increased. The untreated fish had symptoms of vitamin C deficiency. The endogenous liver tissue vitamin C levels were found to reflect well the dietary treatments. After intraperitoneal injection or bath challenge with a virulent strain of Vibrio vulnificus , fish fed with high level vitamin C showed significantly higher survival rate compared with the normal control group. Vaccination with formalin inactivated V. vulnificus significantly enhanced the specific antibody production in fish treated with vitamin C, and completely protected from strong bacterial challenge the groups fed on fish with vitamin C 1500 and 2000 mg/kg diet.

关键词： grouper Epinephelus awoara vitamin C disease resistance immunomodulation pathogen of Vibrio vulnificus

High-throughput simple sequence repeat (SSR) markers development for the kelp grouper (<i>Epinephelus bruneus</i>) and cross-species amplifications for Epinephelinae species

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Advances in Bioscience and Biotechnology 2014年第2期5卷 117-130页

作者： Satoshi Kubota Qi Liu Kanonkporn Kessuwan Nobuaki Okamoto Takashi Sakamoto Yoji Nakamura Yuya Shigenobu Takuma Sugaya Motohiko Sano Susumu Uji Kazuharu Nomura Akiyuki Ozaki Faculty of Marine Science Tokyo University of Ma-rine Science and Technology Tokyo Japan Faculty of Marine Science Tokyo University of Ma-rine Science and Technology Tokyo Japan National Research Institute of Fisheries Science Fi-sheries Research Agency Kanagawa Japan National Research Institute of Aquaculture Fisheries Research Agency Mie Japan National Research Institute of Fisheries Science Fi-sheries Research Agency Kanagawa Japan

The kelp grouper (Epinephelus bruneus), belonging to one of the largest genera among the subfamily Epinephelinae, is a commercially important fish in Japan. There are limited data about the genomics of this species. To provide tools for addressing both population genetics studies and gene mapping, dito pentanucleotide simple sequence repeat (SSR) markers were developed using 454 pyrosequencing. Among the 1466 SSR markers developed, 1244 primer sets produced strong PCR products, of which 905 (72.7%) were polymorphic in kelp grouper. Cross-species utility of the 905 polymorphic SSR markers was tested in four additional Epinephelinae species of Hyporthodus septemfasciatus, Plectropomus leopardus, Epinephelus lanceolatus and Epinephelus coioides. Results revealed that, respectively, 401 (44.3%), 136 (15.0%), 434 (49.0%) and 538 (59.4%) SSRs showed specific polymorphic products. Of these, 40 SSR markers (33 di-, 1 tri- and 6 tetra-nucleotides) showed polymorphism in all species tested. Additionally, three AGAT SSR motifs which accounted for 42.9% of the nondi-nucleotide markers were found in the 40 SSR markers. This indicates that the AGAT SSR motif has a high potential as a highly versatile SSR marker in grouper Epinephelinae. The SSR markers developed in this study can be employed to obtain reliable genetic variability estimates for groupers (Epinephelinae).

关键词： Kelp grouper Epinephelus bruneus Repeat Motif Simple Sequence Repeat (SSR) 454 Pyrosequencing Cross-Species Amplification

Detection of Fish Disease Caused by Iridovirus on grouper （Epinephelus sp.） and Pomfret Star （Trachinotus blochii） with Co-agglutination Method in Tanjungpinang, Indonesia

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Journal of Agricultural Science and Technology(B) 2016年第2期6卷 121-128页

作者： Surya Amanu Dwi Sulistiyono Inyoman Suardana Division of Microbiology Faculty of Veterinary Medicine Gadjah Mada University Yogyakarta 560862 Indonesia Tanjungpinang Fish Quarantine and Inspection Agency Tanjungpinang 29113 Indonesia

Iridovirus infection often causes death and considerable economic losses in the aquaculture industry. This research applies the co-agglutination method that is fast, cheap and accurate in confirming the diagnosis of the cause of an outbreak of illness caused by iridovirus in the field, so that remedial action can be taken quickly and appropriately to minimize the impact of wider losses. Samples were taken from the grouper and pomfret star farms that are experiencing outbreaks of infectious diseases in the months from May to August 2015, Tanjungpinang, Indonesia. The sick and allegedly attacked by iridovirus samples showed abnormal swimming clinical symptoms, weakness and the swollen spleen. The swollen spleen of sick fish created suspension in phosphate buffered saline （PBS） with pH 7.2, and then centrifuged at 8,000 rpm for I0 rain. The supernatant after centrifuge was used as the test sample. On a clean object glass, 50 μL of the supematant was treated with 50 μL kit co-agglutination pre-prepared. The positive results were shown by the agglutination reaction after 10-15 rain, while as a negative control, PBS was reacted with co-agglutination kit that looked homogeneous （no agglutination）. It was showed that the grouper （Epinepkelus sp.） on four farms and pomfret star （Thracinotus blochii） on one farm that experienced an outbreak of infectious disease in Tanjungpinang showed positively infected iridovirus. The same positive iridovirus result was also demonstrated by examination using polymerase chain reaction （PCR） at 570 bp. So, the causative agent of plague on grouper and pomfret star was iridovirus. In addition, the co-agglutination test based on serology is more quick, cheap and accurate.

关键词： Iridovirus grouper pomfret star co-agglutination and polymerase chain reaction.

Advances in the Study on Nutrient Requirements of grouper (Epinephelus sp.): a Review

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Journal of Ocean University of China 2005年第2期4卷 93-98页

作者： LUOZhi LIUYongjian MAIKangsen TIANLixia InstituteofAquaticEconomicAnimals SchoolofLifeSciencesSunYat-senUniversityGuangzhou510275P.R.China FisheriesCollege OceanUniversityofChinaQingdao266003P.R.China

The paper reviews the recent advances in studying grouper nutrition requirement for the development of cost-effective and environmentally friendly artificial diets. It consists of seven parts: protein and amino acid, lipid and essential fatty acid, carbohydrate, vitamin, mineral, alternative protein source, broodstock and larval nutrition. The review provides some basic information for further investigation of nutrient requirements of groupers.

关键词： epinephelus grouper nutrient requirement fish nutrition