点带石斑鱼的亲鱼培育、产卵受精和胚胎发育

在水泥池人工养殖条件下,对人工诱导性逆转的“功能性雄性”、天然雄性及天然雌性点带石斑鱼(Epinephelus malabaricus)亲鱼进行强化培育,自然产卵受精。2002年繁殖季节,共获3cm以上的点带石斑鱼苗145万尾。亲鱼培育、产卵受精和胚胎发育的程序：(1)经过强化培育的亲鱼,自然产卵受精,受精率平均81％,最高97％;(2)成熟的“人工变性雄鱼”的精子头部呈球型,精子头径约3．7—6．1gm,尾长12．4～44．5μm;在水温26℃、盐度3％、pH7．9条件下,精子游动时间最长超过58min,精液中90％精子在31min停止游动,与天然雄亲鱼的精子无异,可作为生产雄亲鱼的来源;(3)在东山岛,产卵期由4月初持续到11月中旬,产卵高潮集中在4月底至5月底、9月底至10月初。在24h内,自然产卵通常从16：30持续到次日1：30;(4)产卵适宜水温为23．5—28．6℃,最适产卵水温为25．5—26．5℃;(5)水温26℃时,受精卵在盐度2．82％～2．96％,的海水中呈悬浮状态,盐度2．82％以下时下沉,盐度2．96％以上时上浮;(6)在水温24．9—27．6℃、盐度3％—3．3％、pH7．6—8．2的情况下,胚胎发育时间为21h。     

湛江棕囊藻对南美白对虾虾苗和多种鱼苗的毒性研究

测定了湛江棕囊藻赤潮海水和棕囊藻(Phaeocystis globosa)对南美白对虾(Penaeus vannamei Boone)虾苗、青石斑鱼(Epinephelus awoara)、鲻鱼(Mugil cephalus)和尖吻鲈鱼(Latescal carifer)鱼苗的毒性。结果表明,棕囊藻对虾苗有一定的毒性,24 h LC50为1.0×109 cells L-1,去除囊泡液后的棕囊藻碎片对虾苗毒性较弱,细胞密度为1.0×109 cells L-1时,24 h虾苗的死亡率仅为10%,赤潮海水对虾苗无毒性;棕囊藻囊泡液对青石斑鱼有一定毒性,24 h LC50为囊泡液占海水总体积的10.9%,赤潮海水对青石斑鱼苗无毒性;棕囊藻对尖吻鲈鱼和鲻鱼鱼苗无毒性。     

An analysis of nucleotide and amino acid variability in the barcode region of cytochrome c oxidase I (cox1) in fishes

The 655 bp cytochrome c oxidase subunit I barcode region of single specimens of 388 species of fishes (four Holocephali, 61 Elasmobranchii and 323 Actinopterygii) was examined. All but two (Urolophus cruciatus and Urolophus sufflavus) showed different cox1 nucleotide sequences (99.5% species discrimination); the two that could not be resolved are suspected to hybridize. Most of the power of cox1 nucleotide sequence analysis for species identification comes from the degenerate nature of the genetic code and the highly variable nature of the third codon position of amino acids. Variation at the third codon position is bimodally distributed, and the more variable mode is dominated by amino acids with four or six codons, while the less variable mode is dominated by amino acids with two codons. The ratio of nonsynonymous to synomymous changes is much less than one, indicating that this gene is subject to strong purifying selection. Consequently, cox1 amino acid sequence diversity is much less than nucleotide sequence diversity and has very poor species resolution power. Fourteen of the 16 amino acid residues recognized as having important functions in the region of cox1 sequenced were completely conserved over all 388 species (and the bovine cox1 sequence), with one fish species varying at one of these sites, and three fish at another site. No significant differences in amino acid conservation were observed between residues in helices, strands and turns. Patterns of nucleotide and amino acid variability were very similar between elasmobranchs and actinopterygians.     

SRIF及CSH对斜带石斑鱼脑垂体生长激素合成和分泌的调控

斜带石斑鱼 (Epinepheluscoioides)属于雌性先成熟、具有性转变的雌雄同体鱼类。生长激素释放抑制因子 (SRIF)是鱼类生长激素 (GH)分泌的主要抑制性调节剂 ,半胱胺 (CSH)可抑制SRIF的作用。本文采用静态孵育系统 ,应用RPA及RIA研究SRIF及CSH对斜带石斑鱼GHmRNA表达及GH分泌的调节。结果显示 ,SRIF能以剂量依存方式抑制斜带石斑鱼脑垂体释放GH ,时间越长作用越强。但SRIF作用 2 4h对GHmR NA水平的影响不显著 ,表明SRIF是斜带石斑鱼GH释放的抑制性调节剂 ,对GHmRNA的表达没有明显影响。较低剂量的CSH (10 -4- 10 -2 mol/L)使斜带石斑鱼的GH释放量增加 ,较高剂量 (10 -1mol/L)的CSH引起的GH增加趋势减缓 ,这种现象可能与较高剂量的CSH不仅抑制下丘脑SRIF的释放 ,同时影响GHRH的释放 ,使得GH的分泌量增幅下降有关 ;无论是较高剂量还是较低剂量的CSH都不能使GHmRNA的水平增加 ,表明CSH只能引起GH的释放量增加 ,不影响GH的合成。GnRH与CSH共同作用引起的GH释放量明显高于CSH单独作用的效应 ,其主要原因是由于GnRH促进GHmRNA的表达所致     

A wireless communication system for determining site fidelity of juvenile dusky groupers Epinephelus marginatus (Lowe, 1834) using coded acoustic transmitters

The value of marine reserves to protect threatened and vulnerable species, such as the dusky grouper (Epinephelus marginatus), is often uncertain because of a lack of knowledge about their ecology and habitat utilisation. To address this issue, a Wireless Communication System for location-based positioning of fish tagged with coded acoustic transmitters was installed to determine site fidelity of juvenile dusky groupers (mean total length = 30.4 cm, range: 20.5–40.0 cm; mean total weight = 470 g, range: 113–950 g) within a marine reserve (Island of Ustica, Sicily, Italy). The system was based on five wireless hydrophones distributed approximately 200 m apart along the coastline of the reserve, each monitoring acoustic signals from transmitters within a reception cell. The hydrophones retransmitted detected signals as a radio signal to a land-based receiver and data logger. Unique identifiers for each hydrophone permitted the identification of the detection cell containing the tagged fish. The wireless hydrophones had partly overlapping detection ranges, dividing the marine reserve into more than five different detection cells. The average detection range was 232 m, but coverage varied from site to site and with time, depending on many factors, such as ambient noise, wave action and signal strength of the transmitters. Despite some limitations, the Wireless Hydrophone System (WHS) combined with manual tracking provided a valuable tool for investigating site fidelity in the dusky grouper. Preliminary home ranges for the fish were estimated by the kernel method (probability level of 95%), based on a limited set of manual tracking data, to be a median 5312 m² (range 1673–18626 m²).     

Molecular characterization and transcriptional analysis of non-mammalian type Toll like receptor (TLR21) from rock bream (Oplegnathus fasciatus)

Toll-like receptors (TLRs) are a large family of pattern recognition receptors, which are involved in triggering host immune responses against various pathogens by detecting their evolutionarily conserved pathogen associated molecular patterns (PAMPs). TLR21 is a non-mammalian type TLR, which recognizes unmethylated CpG DNA, and is considered as a functional homolog of mammalian TLR9. In this study, we attempted to identify and characterize a novel TLR21 counterpart from rock bream (Oplegnathus fasciatus) designated as RbTLR21, at molecular level. The complete coding sequence of RbTLR21 was 2919 bp in length, which encodes a polypeptide of 973 amino acids with a predicted molecular mass of 112 kDa and a theoretical isoelectric point of 8.6. The structure of the deduced RbTLR21 protein is similar to that of the members of typical TLR family, and includes the ectodomain, which consists of 16 leucine rich repeats (LRRs), a transmembrane domain, and a cytoplasmic Toll/interleukin-1 receptor (TIR) domain. According to the pairwise sequence analysis data, RbTLR21 was homologous to that of the orange-spotted grouper (Epinephelus coioides) with 76.9% amino acid identity. Furthermore, our phylogenetic analysis revealed that RbTLR21 is closely related to E. coioides TLR21. The RbTLR21 was ubiquitously expressed in all the tissues tested, but the highest expression was found in spleen. Additionally, upon stimulation with Streptococcus iniae, rock bream iridovirus (RBIV), and Edwardsiella tarda, RbTLR21 mRNA was significantly up-regulated in spleen tissues. Collectively, our findings suggest that RbTLR21 is indeed an ortholog of the TLR21 family and may be important in mounting host immune responses against pathogenic infections.     

性逆转石斑鱼脑垂体差异表达基因克隆的筛选

以17α甲基睾丸酮投喂赤点石斑鱼(Epinephelusakaara),成功地获得了性逆转的有功能的雄鱼。用SMARTcDNA合成和长片段PCR技术构建了性逆转前后脑垂体cDNA文库;用抑制消减杂交技术建立了性反转前后抑制性差减文库。对获得的560个雄鱼脑垂体PCR阳性克隆和350个雌鱼脑垂体PCR阳性克隆进行斑点杂交,共筛选到103个差异表达cDNA片段。对其中29个克隆进行了测序,与GenBank中的已知基因序列进行同源性比较,发现有6个片段为促性腺激素α亚基前体基因(GenBank注册号:AY207430),与同属不同种的石斑鱼促性腺激素α亚基前体基因有97%的同源性;1个片段为生长激素前体(GenBank注册号:AY207431),与同属不同种的石斑鱼生长激素前体基因有99%的同源性;其余22个cDNA片段与GenBank中的序列无明显同源性 。