Does heavy oil pollution induce bacterial diseases in Japanese flounder Paralichthys olivaceus?

As basic research for the effect of heavy oil on the fish immune system, in this study, the number of leukocyte was counted in Japanese flounder Paralichthys olivaceus, after exposure to heavy oil at a concentration of 30 g/8 L for 3 days. To compare the numbers of bacteria in the skin mucus between oil-exposed and control fish, viable bacteria were enumerated by counting colony forming unit (CFU). Compared with 5.79 ± 1.88 × 10⁷ leukocytes/mL in the controls, the exposed fish demonstrated higher counts, averaging 1.45 ± 0.45 × 10⁸ cells/mL. The bacterial numbers of control fish were 4.27 ± 3.68 × 10⁴ CFU/g, whereas they were 4.58 ± 1.63 × 10⁵ CFU/g in the exposed fish. The results suggest that immune suppression of the fish occurred due to heavy oil stressor, and bacteria could invade in the mucus, resulting in the increasing leukocyte number to prevent infectious disease.     

外源激素对养殖牙鲆血浆睾酮和雌二醇含量的影响研究

雄性牙鲆单独注射促黄体激素释放激素类似物(LHRH-A)(每千克质量的注射量,下同)25μg/kg或人绒毛膜促性腺激素(hCG)1 000 IU/kg,血浆睾酮(T)含量在注射后6 h大幅度降低,12h时达到最低值,以后逐渐回升;LHRH-A与hCG联合注射[LHRH-A(12.5μg/kg)+hCG(500 IU/kg)],注射后6 h血浆T含量剧烈下降,24 h时达到最低点,以后缓慢回升;对照组雄鱼血浆T含量随着采样进行而呈现降低趋势.雌鱼单独LHRH-A后,血浆雌二醇(E₂)水平逐渐下降,24 h时达到最低值,30 h后略有回升;单独注射hCG后,血浆E₂含量缓慢下降,30 h达到最低值;LHRH-A与hCG联合注射,注射后6 h血浆E2含量逐渐下降,24 h时达到最低点,30 h时缓慢回升.对照组雌鱼血浆E₂水平也随着采样的进行而逐渐降低.结果表明:单独或联合注射LHRH-A和hCG降低了雌、雄牙鲆血浆E₂和T含量,与对照组结果相同.     

Genetic structure of the common sole (Solea solea) in the Bay of Biscay: Nurseries as units of selection?

Coastal and estuarine areas in the Bay of Biscay are essential nursery habitats for sole (Solea solea). Using three intronic markers, we investigated patterns of genetic differentiation among cohorts of juveniles (0+, 1+, sub-adults) inhabiting four major nurseries in the Vilaine and Loire river estuaries, the Pertuis d'Antioche strait, and the Gironde estuary. Genetic differentiation was not significant for the age 0- and 1-groups, but was among sub-adult samples (θ_WC = 0.017; P = 0.031). Hierarchical analyses of genotypic frequencies by nested multivariate ANOVA using redundancy analysis indicated that northern nurseries of Loire and Vilaine rivers had different patterns of genotypic variation with age than southern nurseries of the Pertuis d'Antioche and Gironde estuaries (F = 1.36; P = 0.007). Bay of Biscay nurseries appeared to be spatially structured between a southern and a northern group that exhibit different genotypic distributions. Genetic drift and gene flow explains this pattern poorly, but a statistical test (P < 0.035) suggested that the metallothionein (MT) locus may be significantly impacted by selection in southern estuaries. As metallothionein is involved in heavy-metal detoxification, and southern nurseries are known to be affected by cadmium, this indicates a possible correlation between observed genotypic distributions and some nursery habitat features in sole. Some selective patterns might also be proposed to explain patterns of linkage disequilibrium observed at amylase loci in three of the four nurseries.     

条斑星鲽早期发育生物学研究——受精卵的形态、生态和卵胚发育特征

条斑星鲽(Verasper moseri)的受精卵为圆球形、分离透明、无油球的悬浮卵。卵膜具粗波纹状的纹络。卵黄均匀,浅黄色或米黄色。卵径平均为1.70 mm±0.02 mm。卵黄囊径1.57 mm±0.025 mm。卵黄间隙狭窄,约为0.11 mm。受精卵的可孵化水温为6.0~11.0℃,最适孵化水温8.0~10.0℃。可孵化盐度为27.00~36.00,最适孵化盐度为33.00~35.00。受精卵在不同盐度的海水中,呈现不同的生态分布特征。在水温8.0~10.0℃,盐度33.00,光照500~6000 lx,pH值7.5~8.4的孵化条件下,受精卵约历时216 h孵化出膜,出膜方式以头部首先破膜为主。优质卵的孵化率可达90%以上。在条斑星鲽胚胎发育过程中,原肠期及原肠期以前的时期对温度的敏感性明显强于原口关闭后各个发育时期。卵胚卵裂期的不同步分裂现象要比其他硬骨鱼类提前,出现在第三次卵裂时期。多细胞期的裂球均一程度较低。在孵化育苗生产中,出现大量带有"空泡"的不正常的卵胚和前期仔鱼。该研究结果为比较鲆鲽鱼类早期发育过程提供了基础参考资料,同时揭示作为冷温性鱼类条斑星鲽受精卵发育过程中出现的特殊规律,从而为该鱼种进一步大规模苗种生产提供了理论依据。     

Analysis of new microsatellite markers developed from reported sequences of Japanese flounder <Emphasis Type="Italic">Paralichthys olivaceus</Emphasis>

The expressed sequence tags (ESTs) of Japanese flounder, Paralichthys olivaceus, were selected from GenBank to identify simple sequence repeats (SSRs) or microsatellites. A bioinformatic analysis of 11111 ESTs identified 751 SSR-containing ESTs, including 440 dinucleotide, 254 trinucleotide, 53 tetranucleotide, 95 pentanucleotide and 40 hexanucleotide microsatellites respectively. The CA/TG and GA/TC repeats were the most abundant microsatellites. AT-rich types were predominant among trinucleotide and tetranucleotide microsatellites. PCR primers were designed to amplify 10 identified microsatellites loci. The PCR results from eight pairs of primers showed polymorphisms in wild populations. In 30 wild individuals, the mean observed and expected heterozygosities of these 8 polymorphic SSRs were 0.71 and 0.83 respectively and the average PIC value was 0.8. These microsatellite markers should prove to be a useful addition to the microsatellite markers that are now available for this species.     

Cloning of the cDNA encoding adenosine 5′-monophosphate deaminase 1 and its mRNA expression in Japanese flounder Paralichthys olivaceus

AMP deaminase catalyzes the conversion of AMP into IMP and ammonia. In the present study, a full-length cDNA of AMPD1 from skeletal muscle of Japanese flounder Paralichthys olivaceus was cloned and characterized. The 2 526 bp cDNA contains a 5’-UTR of 78 bp, a 3’-UTR of 237 bp and an open reading frame (ORF) of 2 211 bp, which encodes a protein of 736 amino acids. The predicted protein contains a highly conserved AMP deaminase motif (SLSTDDP) and an ATP-binding site sequence (EPLMEEYAIAAQVFK). Phylogenetic analysis showed that the AMPD1 and AMPD3 genes originate from the same branch, but are evolutionarily distant from the AMPD2 gene. RT-PCR showed that the flounder AMPD1 gene was expressed only in skeletal muscle. QRT-PCR analysis revealed a statistically significant 2.54 fold higher level of AMPD1 mRNA in adult muscle (750±40 g) compared with juvenile muscle (7.5±2 g) (P<0.05). HPLC analysis showed that the IMP content in adult muscle (3.35±0.21 mg/g) was also statistically significantly higher than in juvenile muscle (1.08±0.04 mg/g) (P<0.05). There is a direct relationship between the AMPD1 gene expression level and IMP content in the skeletal muscle of juvenile and adult flounders. These results may provide useful information for quality improvement and molecular breeding of aquatic animals.     

牙鲆选育F3代家系的建立及遗传效应分析

牙鲆是太平洋西岸海水养殖、捕捞和自然增殖的主要鱼类,但是人工养殖病害多、生长慢,针对这一问题,从2003年开始开展了牙鲆选择育种。本文利用筛选和培育的F1、F2代牙鲆家系核心群体,构建了F3代家系52个,在生长到196d时,对家系1 560尾鱼苗全长、体宽、体质量、全长日增量和体重日增量5个生长性状进行测量和计算,获得7 800个数据。采用"加性-显性"线性模型、最小范数二阶无偏估计法(MINQUE)对以上生长性状的边际平均值、加性方差分量、显性方差分量、随机方差分量、狭义遗传力、广义遗传力、亲本加性效应、杂交随机效应等遗传参数进行了评估。结果显示:1202、1206、1226和1211家系以上生长性状的边际平均值同时显著高于其他家系。以上5个生长性状的加性方差分量比率为(0.099±0.01)~(0.24±0.02),显性方差分量比率(0.30±0.02)~(0.31±0.02),随机方差分量比率(0.48±0.02)~(0.61±0.01),5个性状狭义遗传力0.099~0.24,广义遗传力0.38~0.54,以上遗传参数均达极显著性水平(P0.01)。综合比较18个亲本5个性状的加性效应预测值,发现F0917、F0908、F0905、F0915、F09125和KS(韩国牙鲆)亲本在5个性状上都具有极显著的正向效应(P0.01),其他亲本5个性状都不同程度的表现为显著(P0.05)或极显著(P0.01)的负向加性效应。对42个杂交组合显性随机效应进行预测,D(2×6)、D(2×18)、D(3×4)、D(7×12)、D(7×14)和D(8×13)6个杂交组合在5个性状上都表现为极显著(P0.05)正效应,利用这些杂交组合建立家系将促进牙鲆以上5个性状的生长。本文研究结果,为牙鲆优良家系的建立及新品种的培育初步筛选出了性状优良的亲本、杂交组合和家系,同时为牙鲆进一步选择育种提供了理论和技术依据。     

Spatial and seasonal patterns in sighting rate and life-history composition of the white shark Carcharodon carcharias at Mossel Bay,South Africa

White sharks Carcharodon carcharias aggregate at specific times of the year at localities along the South African coast. At Mossel Bay, on the southern Cape coast, four sites were sampled (Seal Island, Hartenbos, Kleinbrak and Grootbrak) to investigate spatial and seasonal patterns in relative abundance and life-history composition. These are known aggregation sites within the bay, each having particular physical and/or biological characteristics. Sightings-per-unit-effort data were collected from February to December 2008–2010. Sighting rates demonstrated significant seasonal and interannual variation at the four sites. The highest mean sighting rate was recorded at Seal Island and the lowest at Hartenbos, which might be a consequence of differences in prey availability. The greatest interannual variability was recorded at Kleinbrak, followed by Seal Island, with little variability at Grootbrak and Hartenbos. White sharks appeared to concentrate at Grootbrak and Kleinbrak in summer and autumn, at Seal Island in winter, and at Hartenbos and Seal Island in spring. All life-history stages were present year-round but their occurrence was influenced significantly by season (p < 0.05), although not site. Few adults (325–424 cm total length) were seen, with the highest frequency being in spring, whereas that of young-of-the-year (≤174 cm) was in autumn. Juveniles (175–324 cm) constituted 78% of the animals sighted, indicating that Mossel Bay is an important aggregation site for this life-history stage.