Takifugu bimaculatus is a cultured marine teleost fish, distributed ranging from the southern Yellow Sea to the South China Sea. Understanding the mechanism of sex determination is essential for sex control breeding biotechnologies of T. bimaculatus. However, there lacks a rapid and accurate sex specific marker for the T. bimaculatus. In this study, whole genome resequencing was conducted on 71 individuals (including 49 females and 22 males), 331,190 high-quality polymorphic loci (34,435 INDEL, 296,755 single-nucleotide polymorphisms (SNPs)) were obtained through genome-wide association analysis (GWAS) by GEMMA (Genome-wide Efficient Mixed Model Association algorithm). 1403 loci on chromosome 7 were identified as sex significantly associated loci (P < 1.51 x 10(-7)), and a 12 Mb sex-associated region on Chr7 was narrowed down by F-st, scanning. It was worth noting that the SNP, SNP1100879 on Chr7 at position 8,764,381 was identified to be sex determination in T. bimaculatus. 13 pairs of primers for sex identification Marker-F/R were designed and it was validated by PCR. However, only one pair of primers (SNP-7F/R) was sex-linked, in which 95% of female genotypes are homozygous CC and 88% of male genotypes are heterozygous CT, suggesting a XX/XY sex determination system in T. bimaculatus. In addition, 12 sex-related candidate genes were identified around 306 SNPs, including amhr2, dzip1, patz1, brdt, hmga2, gli1, inhbb, mmp19, wnt2b, taf4 and so on. Because of the significant difference of gonadal weight (GW) and gonadosomatic index (GSI) between females and males of T. bimaculatus, GWAS on GW and GSI of different genders were also conducted independently. Besides, we identified 11 SNPs, located on Chr7, Chr18, Chr21, and Chr2, showing genome-wide significant association with GW and GSI and five genes including cyp27b1, hexb, inhba, cgba and shbg, involved in oogenesis, ovarian follicle development, and regulation of estradiol secretion. Then, we performed the gene ontology (GO) and KEGG signaling pathway (KEGG) and Protein-protein interaction (PPI) network analyses of the candidate genes. amhr2, gli1, ihbb and dzip1 genes were identified as hub genes. In the present study, a sex-linked marker of T. bimaculatus was identified for the first time using whole-genome resequencing technology, and its limited versatility provided important clues for deciphering the sex determination mechanism in T. bimaculatus.

双斑东方鲀(Takifugu bimaculatus)隶属鲀形目、鲀科、东方鲀属,分布于南海、东海和黄海南部,为近海暖温性底层鱼类,主要以贝类、甲壳类等无脊椎动物和小鱼为食.双斑东方鲀肉质鲜嫩可口,营养价值高,高蛋白低脂肪,目前已在福建省漳州市沿海开展人工养殖. 福建省水产研究所东方鲀种业创新与产业化工程项目组在福建省南部沿海池塘采用鱼虾生态混养方式,进行双斑东方鲀健康养殖示范.根据对虾的养殖周期,分为春夏季、秋冬季两个阶段进行养殖.春夏季养殖池塘投放一冬龄双斑东方鲀苗种,混养凡纳滨对虾,收捕凡纳滨对虾之后,进行清池消毒.池塘重新进水,经过春夏季养殖的双斑东方鲀放回原池进行秋冬季养殖,并混养日本囊对虾.次年1月～3月双斑东方鲀、日本囊对虾捕捞销售.该分段养殖模式可有效提高池塘利用率,降低养殖风险,可为双斑东方鲀养殖产业化发展提供一种新模式.

Myostatin (MSTN) is a negative regulator of muscle growth, which restrains the proliferation and differentiation of myoblasts. To understand the role of two mstn genes of Takifugu bimaculatus, the full-length cDNAs of 1131 bp Tbmstn1 and 1,080 bp Tbmstn2 were obtained from the T. bimaculatus’ genomic database, which encodes 376 and 359 amino acids, respectively. The results of qRT-PCR showed that Tbmstn1 was expressed in the eye, kidney, spleen, skeletal muscle, gill, and brain, and the expression level in the skeletal muscle was extremely significantly higher than in other examined tissues. Tbmstn2 was expressed in the skin, skeletal muscle, gill, and brain, and had the highest expression in the skeletal muscle, followed by expression in the brain. Meanwhile, in different stages of embryonic development, the expression of Tbmstn1 started from the gastrula stage. Its expression in the eye-pigment formation stage and hatching stage was significantly higher than that in other stages. The Tbmstn2 was expressed in all examined embryonic stages with different levels, and the highest expression was detected in the eye-pigment formation stage. These results suggested that Tbmstn1 and Tbmstn2 may involve in the development of skeletal muscle, and Tbmstn2 may be related to the formation of nervous system.

Pufferfish are ideal models for vertebrate chromosome evolution studies. The yellowbelly pufferfish, Takifugu flavidus, is an important marine fish species in the aquaculture industry and ecology of East Asia. The chromosome assembly of the species could facilitate the study of chromosome evolution and functional gene mapping. To this end, 44, 27 and 50 Gb reads were generated for genome assembly using Illumina, PacBio and Hi-C sequencing technologies, respectively. More than 13 Gb full-length transcripts were sequenced on the PacBio platform. A 366 Mb genome was obtained with the contig of 4.4 Mb and scaffold N50 length of 15.7 Mb. 266 contigs were reliably assembled into 22 chromosomes, representing 95.9% of the total genome. A total of 29,416 protein-coding genes were predicted and 28,071 genes were functionally annotated. More than 97.7% of the BUSCO genes were successfully detected in the genome. The genome resource in this work will be used for the conservation and population genetics of the yellowbelly pufferfish, as well as in vertebrate chromosome evolution studies.

刺激隐核虫病是双斑东方鲀养殖生产中的主要致死性寄生虫病,为安全、有效地防治刺激隐核虫病,开展了硫酸铜对双斑东方鲀幼鱼的急性毒性试验及硫酸铜对双斑东方鲀刺激隐核虫病的治疗效果试验.急性毒性试验结果显示,硫酸铜对双斑东方鲀幼鱼24、48、72、96 h半致死质量浓度分别为7.94、7.12、5.32、4.44mg/L,其安全质量浓度为1.72mg/L.刺激隐核虫病治疗试验结果表明,1.0、1.5mg/L的硫酸铜质量浓度对病鱼治疗有效,药浴后刺激隐核虫虫体活力下降,第3d部分虫体解体或脱落,药浴5d治愈;0.5mg/L硫酸铜对病鱼治疗效果不佳,病情继续发展,第8d病鱼死亡.临床上建议使用硫酸铜质量浓度1.0 mg/L治疗双斑东方鲀刺激隐核虫病.本研究对其他鱼类刺激隐核虫病及其纤毛虫病治疗也具有一定的借鉴意义.

利用16S rRNA高通量基因测序技术研究了双斑东方鲀(Fugu bimaculatus)循环水养殖系统养殖水体细菌种群结构特征,并比较养殖患病(溃疡症)的双斑东方鲀的水体细菌群落结构与健康的之间的差异,探讨细菌群落结构与双斑东方鲀细菌性病害发生的相关性.研究结果表明,双斑东方鲀循环水养殖水体细菌群落的优势细菌门为变形菌门(Proteobacte-ria)、 拟杆菌门(Bacteroidetes),优势细菌纲为γ-变形菌纲(Gammaproteobacteria)、 黄杆菌纲(Flavobacteriia)、 α-变形菌纲(Alphaproteobacteria),优势细菌目为黄杆菌目(Fla-vobacteriales)、 硫发菌目(Thiotrichales)、 红细菌目(Rhodobacterales)、 交替单胞菌目(Al-teromonadales)、 弧菌目(Vibrionales),优势细菌科为黄杆菌科(Flavobacteriaceae)、 硫发菌科(Thiotrichaceae)、 红细菌科(Rhodobacteraceae)、 弧菌科(Vibrionaceae)、 交替假单胞菌科(Pseudoalteromonadaceae).患病的和健康的循环水养殖水体细菌群落种类组成相似度高,细菌群落多样性指数无较大差异,在同一分类水平上各细菌群落丰度占比差异显著;二者核心微生物(属)的差异明显,健康的循环水养殖水体核心细菌群落以Polaribacter属为主;患病的循环水养殖水体核心细菌群落以弧菌科的发光杆菌属(Photobacterium)和弧菌属(Vibrio)为主,而发光杆菌属和弧菌属的多数种类属于病原菌或条件致病菌.双斑东方鲀患病(溃疡症)与养殖水体细菌群落的弧菌属丰度占比高有关.该研究结果对双斑东方鲀循环水养殖细菌性病害的监测、 预警、 诊断及管理控制具有重要的意义.

在水温(27±1)℃、 盐度25±1、PH值为7.90~8.10、 溶解氧>6.00 mg/L条件下,采用静水法进行了高锰酸钾、 聚维酮碘、 甲醛、 戊二醛、 敌百虫、 乙酸铜等6种药物对双斑东方鲀幼鱼急性毒性试验.结果表明:高锰酸钾、 聚维酮碘、 甲醛、 戊二醛、 敌百虫和乙酸铜对双斑东方鲀幼鱼半致死浓度(24、48、72、96 h)分别为(1.88、1.84、1.76、1.75 mg/L)、(23.30、22.55、20.34、19.50 mg/L)、(11.27、7.06、6.38、5.57 mg/L)、(9.25、9.03、7.03、5.48 mg/L)、(16.85、14.19、10.23、7.92 mg/L)和(10.54、8.78、6.13、5.46 mg/L),安全浓度分别为0.53、6.34、0.83、2.58、3.02和1.83 mg/L.双斑东方鲀幼鱼对6种药物耐受能力的强弱依次为:聚维酮碘>敌百虫>戊二醛>乙酸铜>甲醛>高锰酸钾.实际生产上应严格按照药物安全浓度控制药物的使用.

采用福建省水产研究所研制的“河豚毒素半定量快速检测试剂盒”,在双斑东方鲀人工繁殖及苗种培育过程中,对不同繁殖群体亲种,不同繁殖群体交配所得受精卵、胚胎及仔稚幼鱼的河豚毒素进行跟踪检测.结果表明:不同繁殖群体亲鱼组织器官毒性差异大,亲鱼眼珠、精巢为阴性,皮肤、肌肉、肝脏、卵巢呈现不同阳性检出率,分别为5.4％、16.2％、40.5％、40.5％;养殖繁殖群体亲鱼皮肤、肝脏为阳性,野生繁殖群体亲鱼肝脏和雌性性腺河豚毒性阳性检出率比养殖繁殖群体亲鱼高;野生群体雌性亲鱼与野生群体雄性亲鱼、养殖群体雄性亲鱼交配组受精卵河豚毒素均为阳性,早期胚胎为阳性,后期胚胎为阴性;养殖群体雌性亲鱼与野生群体雄性亲鱼、养殖群体雄性亲鱼交配组受精卵、胚胎河豚毒素皆为阴性;野生群体雌性亲鱼与野生群体雄性亲鱼交配组仔稚鱼河豚毒素为阳性,早期幼鱼为阳性,后期幼鱼为阴性;野生群体雌性亲鱼与养殖群体雄性亲鱼交配组仔鱼河豚毒素为阴性,稚鱼、早期幼鱼为阳性,后期幼鱼为阴性;养殖群体雌性亲鱼与野生群体雄性亲鱼、养殖群体雄性亲鱼交配组仔稚鱼河豚毒素皆为阴性.

In the southeast coastal area of China, tetrodotoxin poisoning often occurred for eating puffer fish. Currently main detection methods of tetrodotoxin are mouse assay and enzyme linked immunosorbent assay ( ELISA) etc. The tetrodotoxin detection methods presently need much more effort in time and cost, and can not meet the market requirements nowadays. We developed a tetrodotoxin colloidal gold immunochromato-graphic test kit, which the lowest detection limit was 100 ng/mL. The logic four parameter curve fitting, var-iation coefficient analysis, and standard test methods were used to test the stability, sensitivity, the fitting degree of the trend line, and detection range of homemade detection card in tetrodotoxin quantitative detec-ting. The results showed that the variation coefficient of homemade tetrodotoxin fluorescence immuno-chrom-atographic testing card was 0. 124; correlation coefficient r2 value was 0. 995 5; the detection range was 2~600 ng/mL. The testing card can meet the need of tetrodotoxin detection.

为获得具有杂交优势东方鲀苗种,开展了双斑东方鲀与红鳍东方鲀杂交组、对照组人工繁殖及育苗;正交组(SH)、反交组(HS)胚胎发育观察;杂交组、对照组幼鱼耐高温试验.结果表明:正交组、反交组、双斑东方鲀子一代对照组(SB)、红鳍东方鲀子一代对照组(HQ)卵受精率分别为95.6％、93.5％、96.0％、89.5％,受精卵孵化率分别为92.2％、80.6％、89.4％、79.9％,室内育苗成活率分别为28.4％、18.7％、27.5％、19.2％;正交组(SH)池塘生态培育成活率为18.3％;在18.0～20.0C水温条件下,正交组(SH)受精卵经132 h孵出仔鱼,反交组(HS)受精卵经164 h40 min孵出仔鱼;在28.0 ～35.0℃水温条件,反交组(HS)、对照组(HQ)幼鱼不能存活,正交组(SH)、对照组(SB)幼鱼正常生长;杂交子代生物学特性与母本相近,正交组(SH)可在福建、广东、海南等南方沿海地区推广养殖.

为开展黄鳍东方鲀(Fugu xanthopterus)批量全人工育苗、胚胎及仔稚幼鱼发育特征研究,作者使用地欧酮(DOM)、促黄体释放激素类似物(LRH)及绒毛膜促性腺激素(HCG)对性成熟黄鳍东方鲀进行人工催产。结果表明黄鳍东方鲀亲鱼效应时间为39~60h,获产率为55.0%~92.5%；卵受精率为35.5%~84.7%,3年产卵7400 g；受精卵在海水水温17.0~23.2℃、盐度21~30、pH8.0~8.2人工条件下孵化,孵化率为33.6%~91.7%,孵出仔鱼499万尾；2005年、2006年经前期培育,共培育出黄鳍东方鲀后期仔鱼394.5万尾,成活率为68.2%~94.3%；室内水泥池后期培育出平均全长2.6 cm±0.2 cm幼鱼3100尾,成活率为15.5%；池塘生态培育出平均全长3.7 cm±0.5 cm幼鱼53.3万尾,成活率为11.2%~13.8%。黄鳍东方鲀受精卵为沉性黏性卵,近椭球形,乳白色,卵膜厚且不透明,卵径0.9~1.0mm,在海水水温18.2~22.2℃条件下,3 h分裂为2细胞；3 h 20 min 4细胞；3 h 45 min 8细胞；10 h 10 min多细胞；17 h 30 min囊胚出现；28 h原肠期；57 h 15 min胚体形成期；69 h 20 min心跳出现；130 h 50 min出膜期；144 h孵出仔鱼。初孵仔鱼全长2.2~2.4 mm,体圆而粗短,尾部细长、透明；孵出后第20天,胸鳍、臀鳍、背鳍基本形成,进入稚鱼期；孵出后第45天,上下颌门齿发达,各鳍发育完整,小刺基本形成,进入幼鱼期。

To provide basic data for the economy utilization of interspecific hybridization of grouper species,we conducted hybridization between Epinephelus akaara( ♀) and Epinephelus moara( ♂) by artificial insemination. The morphological features and growth of the hybrid F1 were analyzed. The growth trend in hybrid F1 was close to the Epinephelus moara,the body morphological features and color in hybrid F1 were close to Epinephelus akaara. The hybrid F1 could live through winter in Fujian. These results suggested that the hybrids of Epinephelus akaara( ♀) × E. moara( ♂) had practical value in aquaculture.

In this study artificial crossbreeding between Fugu xanthopteru and Fugu bimaculatus was made.The In-ter-Simple Sequence Repeats (ISSR)technology was applied to analyze the genetic diversity between Fugu xan-thopterus,Fugu bimaculatus and their hybrid progeny (F1).The results showed that there were some particular IS-SR loci among Fugu xanthopterus,Fugu bimaculatus and their reciprocal hybrids.The genetic diversity index (H′) of parent was 0.314 8 and 0.276 5 while between their reciprocal hybrids was 0.336 2 and 0.325 7.The genetic distance (D)between Fugu xanthopterus and Fugu bimaculatus was 0.895 while the genetic distance between the reversed hybrid population and Fugu bimaculatus was 0.169.This experiment indicated that the effect of Fugu bi-maculatus on hybrid progeny was much more significant than Fugu xanthopterus.

ISSR analysis was used to investigate the genetic relationships of five Nile Tilapia populations from different areas called GF,NLⅠ,NLⅡ,NLⅢ and NLⅣ in this paper.Among 81 ISSR primers,15 primers were able to generate highly reproducibly polymorphic DNA fragments.The genetic trees were constructed based on ISSR amplification finger printings using unweighted pair group method for arithmetic averages analysis(UPGMA).According to the tree,Population NL Ⅲ and NL Ⅳ formed one branch first,then grouped with population GF.Population NLⅠand NLⅡ formed another branch.This result showed that population GF separated from other Nile Tilapias populations;The 500 bp band amplified by primer UBC835 could distinguish population GF from other Nile Tilapia populations.

Growth and survival rates of Octopus vulgaris paralarvae feeding on rotifers,enriched artemia nauplii,enriched Moina mongolica,Daphnia-like larvae of Portunus trituberculatus were tested.The result showed that rotifer was not appropriate for Octopus vulgaris paralarvae as diet;the effect sequence of diets on growth and survival rates of octopus vulgaris paralarvae in constant temperature test was mix enriched artemia nauplii>mix enriched Moina mongolica > artemia nauplii enriched by Chlorella>Moina mongolica enriched by Chlorella;the effect sequence of diets on growth and survival rates of octopus vulgaris paralarvae at room temperature test was daphnia-like larvae of Portunus trituberculatu+artemia nauplii > artemia nauplii enriched by fish oil > mix enriched artemia nauplii.It may be considered that the "daphnia-like larvae of Portunus trituberculatu + artemia nauplii" was the better food for the paralarvae of Octopus vulgaris.

Hatching,abnormal,growth and survival rate of the fertilized eggs of Nothobranchius guentheri whitc were carried by Shenzhou 7 spacecraft were studied.The results indicated that the hatching and abnormal rate were no significant difference between the spaceflight group(99.3% and 16.8%) and ground group(97.2% and 10.4%);but the growth rate of male fish from spaceflight group was significant higher(0.094g/d) than that of ground group(0.059g/d),leading to the significant bigger of the male fish from spaceflight group.The survival rate of spaceflight group(66.7%) was higher than the ground group(47.9%).It was concluded that there was a higher growth and survival rate of Nothobranchius guentheri fertilized eggs after space flight.

This paper reported the result of the embryonic development in Seriola aureovittata.The incubation conditions were: water temperature 23℃~25℃,salinity 24~27,pH 8.0~8.3.The fertilized eggs became 2-cell at about 40min,blastula stage at about 3h and 20min,gastrula stage at about 6h and 55min and the larval hatched out at about 41h and 30min after fertilization.

As one new kind of burgeoning marine culture type in Fujian province,the octopus has a high nutritional value,a commodity value and industrial culture prospects.This article gives a general introduction to the classification,distribution,zoology habits,bio-characteristics,artificial reproduction and culture of Octopus.

The paper reports the process and results of studies on artificial propagation and larva-rearing of Fugu flavidus done in Fujian Fisheries Research Institute in 2004~2005.In spring,when the water temperature is above 20℃,mature parent fishes are induced to spawn by injecting DOM,LHRH or HCG,mixedly or respectively.The dosage of hormone depends on the sexual maturity of the parent fishes.The zygotes incubated under the condition of seawater salinity 20.0~30.0 and temperature 20.0~25.0℃.It took 144~156 hours for larvae to hatch.During the studies,about 60.8×106 larvae were obtained.,and all larvae were raised in outdoor ponds or indoor concrete pools.At the end,about 16.85×106 fingerlings,which mean total length is 4.43 cm and mean body weight is 1.98 g.The survival rate is 27.7%.The key technologies on parent fishes raised,inducing to spawn,incubation and pre-larvae and post-larvae raising also have been discussed in the present paper.