Feeding behaviour of Japanese flounder larvae under laboratory conditions

Tank–reared Japanese flounder larvae, Paralichthys olivaceus , had a major feeding peak in the morning and a secondary peak in the afternoon throughout the larval development, with light being the primary factor regulating their feeding activity. The larvae consumed rotifers in preference to Artemia for up to 10 days, after which the food preference shifted to Artemia . Feeding rates of the larvae prior to 10 days post–hatch depended on prey density, but in the old larvae, feeding rates were independent of prey density. Maximum feeding rate occurred at 19° C. The occurrence of the attack posture, after its onset at first feeding (2 days post–hatch), increased up to 25 days, began to decrease when the larvae prepared to settle down, then disappeared after settlement. The occurrence frequency of the attack posture was positively related to fish density, but inversely related to starvation duration, and occurred most frequently at 19° C. This posture depended on prey density in larvae prior to 10 days post–hatch, but became independent of prey density as the larvae developed. It was obvious that, for flounder larvae, attack posture was a behavioural character closely related to feeding and subject to larval development and environmental factors.     

Genomic Bacterial Artificial Chromosome Library of the Japanese Flounder Paralichthys olivaceus

We have constructed a genomic bacterial artificial chromosome (BAC) library from homozygous cloned Japanese flounder Paralichthys olivaceus using the pBAC-lac vector. This BAC library consists of about 49,100 clones and is deposited in 128 microtiter plates with 384 wells. The average size of inserted DNA was calculated to be 165 kb. The BAC library was determined to cover 9 times the Japanese flounder haploid genome. The Japanese flounder genomic BAC library will be useful for gene isolation as well as quantitative trait loci (QTL) analysis. Received March 1, 2000; accepted May 29, 2000.     

Effects of starvation on morphometric characteristics of olive flounder, <Emphasis Type="Italic">Paralichthys olivaceus</Emphasis>

An experiment was conducted for 12 weeks to determine the effect of feeding and starvation on truss and classical parameters in the external body of olive flounder, Paralichthys olivaceus. There was an increase in the truss dimension of body depth in the trunk region of the fed group at the end of the experiment (P < 0.05). In the olive flounder, the trunk region dimensions, including body depth measurements, are likely to be compromised by variability related to differences in the feeding of fish from different habitats. Classical dimensions in relation to the anterior–posterior body axis decreased and classical dimensions of head characteristics increased upon starvation but decreased upon feeding (P < 0.05). These results suggest that these morphometric parameters may be a useful index of nutritional status in olive flounder.     

Physiological responses of the olive flounder, <Emphasis Type="Italic">Paralichthys olivaceus</Emphasis>, to a series stress during the transportation process

The main objective of this investigation was to examine physiological changes occurring within the olive flounder, Paralichthys olivaceus, in the presence of continuous stress factors. The experiment gave continuous stress, and each stress step is as follows: water level reduction (S1), selection process (S2), confinement (S3), air exposure (S4), transportation (S5), and storing in volume 0.5-t (S6) and 50.0-t (S7) rearing tanks for 24 h after transportation. The cortisol concentration showed a trend to continuously rise in response to consecutive stress from transportation from 7.4 ± 0.6 ng/ml in an experimental opening. The concentration showed the highest level, 25.3 ± 4.4 ng/ml, after confinement stress. Glucose concentration in S3 and S4 were increased significantly to 71.0 ± 13.0 and 78.7 ± 7.0 mg/dl, respectively (P < 0.05). Lactic acid was increased significantly from 0.5 ± 0.1 mM (S0) to 9.0 ± 1.2 mM (S4), but it did not recover until 24 h (7.0 ± 1.0 mM) after transportation (S6 and S7) (P < 0.05). Some contents displayed recovery within 24 h, but a longer time would be required for normal physical metabolism to resume after fish experienced stress. Therefore, it would be endangering the survival of this species if the transport were repeated within a 24-h period.     

牙鲆碱性磷酸酶cDNA序列分析与蛋白质高级结构预测

为研究碱性磷酸酶(EC 3.1.3.1; alkaline phosphatase,ALP)在牙鲆(Paralichthys Olivaceus)发育和变态中的作用,采用RACE的方法克隆了牙鲆ALP基因cDNA全长,通过生物信息学分析了核苷酸序列并进行蛋白结构预测. 结果表明,牙鲆ALP cDNA全长为1 811bp,能编码476个氨基酸的蛋白质,分子量为52 293.1,等电点为7.67. 编码区核苷酸GC含量在ALP同源基因中差异比较大,脊椎动物明显高于非脊椎动物和细菌. 分子系统分析显示,牙鲆ALP和青黑斑河豚(Tetraodon nigroviridis)、斑马鱼（Danio rerio）的组织非特异性ALP有较高的同源性,分子进化树和物种进化树是一致的. 在蛋白序列中的一些重要的功能位点,包括金属离子结合位点、N糖基化位点和丝氨酸磷酸化位点等表现了较高的保守性. 牙鲆ALP和人胎盘ALP(PALP)在蛋白序列上有43%的相似性,其3D结构非常接近.通过氨基酸空间位置比较发现,牙鲆ALP中141和203位半胱氨酸对应于人PALP的121和183位半胱氨酸,推测能形成一个二硫键. 在两者酶活性中心,3个金属离子结合的氨基酸残基非常保守,Zn离子周围的9个氨基酸中有2个不同;Mg离子周围的7个氨基酸也只有2个不同,包括一对类似的丝氨酸155和苏氨酸175.     

Genetic differentiation among common and selected hatchery populations of flounder: Evidence from RAPD markers

Randomly amplified polymorphism DNA (RAPD) analysis was performed in order to evaluate one common population and two selected hatchery populations of flounder Paralichthys olivaceus. A total of 90 individuals belonging to three populations were screened using 12 different RAPD primers. A total of 71 loci were produced in the three studied populations, and 43.5%, 42.4% and 42.2% of these loci were polymorphic over all the genotypes tested in common, susceptible and resistant populations, respectively. The number of loci detected by single primers ranged from four to eight. The average heterozygosites of common, susceptible and resistant populations were 0.112, 0.103 and 0.111, respectively. Compared with the common population, the two selected hatchery populations, susceptible and resistant, showed some genetic differences including a smaller number of loci, a smaller number of polymorphic loci, a smaller number of genotypes and a smaller Shannon index. The present study confirmed that it is important to detect the genetic variability of the selected hatchery population for the conservation of natural flounder resources.     

Effects of phenanthrene on growth and basic physiological functions of the olive flounder, Paralichthys olivaceus

Flounder were exposed to waterborne phenanthrene (0.5, 1 and 2 μM) for 4 weeks to test effects of waterborne phenanthrene on growth and hematological properties of the olive flounder (Paralichthys olivaceus). The average weight gain (WG) of flounder was significantly decreased in fish exposed to phenanthrene at 2.0 μM for 2 weeks, whereas WGs of fish treated by 1.0 and 2.0 μM phenanthrene for 4 weeks were significantly decreased. However, hepatosomatic index (HSI) and condition factor (CF) of flounder were not significantly affected by phenanthrene exposure. Red blood cell (RBC) count, hemoglobin (Hb) concentration, hematocrit (Ht), the mean corpuscular hemoglobin (MCH) and the mean corpuscular hemoglobin concentration (MCHC) mean levels were decreased with an increase in exposure time of phenanthrene to the fish, but the level of the mean corpuscular volume (MCV) was increased. Plasma bilirubin concentrations were significantly increased following exposure to waterborne phenanthrene (2.0 μM) for 2 and 4 weeks; however, there were no significant changes in plasma total cholesterol in fish of all treated groups compared to control. The phenanthrene-exposed groups (≥1.0 μM) showed significantly higher mean plasma lysozyme activity. Kidney lysozyme activity of fish exposed to phenanthrene (≥1.0 μM) was also significantly higher than that of control fish. The central finding from these data is that olive flounder exposed to waterborne phenanthrene at more than 1.0 μM are likely to experience negative impacts on fish health and basic physiological functions.     

Morphological development and allometric growth patterns in hatchery-reared California halibut larvae

Morphological development, allometric growth and behaviour of hatchery-reared California halibut Paralichthys californicus were studied from hatching to metamorphosis (42 days post hatch, dph) at 187° C. Mean standard length ( L _S) of larvae and juveniles increased from 2.1 mm at hatching to 10.5 mm at metamorphosis with the increase in length being approximately linear. Stages of morphological development were described using the alphabetic staging (A–I) used for other flatfish species. Organogenesis and differentiation were more rapid and complex in yolk-sac (hatching, stage A–3 dph, stage B), preflexion (3–19 dph, stages B–C), and flexion larvae (from 20 to 23 dph, stages D–E), as larvae developed most of their sensory, feeding, respiratory and swimming systems. After notochord flexion at 24–25 dph (stage F), most morphological changes were related to the progressive transformation from a bilateral symmetrical larva to an asymmetrical benthic juvenile (42 dph, stages G–I).     

日本牙鲆主要组织相容性复合体DAB等位基因的多态性

根据牙鲆(Paralichthys olivaceus)主要组织相容性复合体(MHC)DAB基因序列设计特异性引物,在日本牙鲆基因组中扩增了包括DAB基因完整外显子2和内含子1在内的长度为408 bp的DNA片段.对该片段克隆测序后,发现了37个MHC-DAB等位基因,各等位基因的频次以及各主型中亚型数目分布都不平衡.在第二外显子273 bp核苷酸序列中有50个位点发生变异,核苷酸多样性PI值为0.0618,在编码的氨基酸序列中存在多态变异位点30个,其中简约信息位点29个,单变异位点1个.非同义替代与同义替代的比率在抗原结合区(PBR)和非PBR编码区分别为10.0和1.62,分析表明正向选择机制可能是产生牙鲆MHC-DAB多态性的主要因素.估计的核苷酸的转换和颠换数随着遗传距离的增加而增加.各等位基因间的系统发生关系表明,19个主型分为两个群.对氨基酸组分和密码子的偏倚性以及分布频率的分析表明各同义密码子的使用频率不均衡.本研究为牙鲆MHC-DAB基因的遗传进化和分子标记辅助育种的研究提供了理论基础.     

Effects of a muscle‐infecting parasitic nematode on the locomotor performance of their fish host

The southern flounder Paralichthys lethostigma, host to the nematode Philometroides paralichthydis that is embedded in place of the inclinator muscles of the dorsal and anal fin elements, is hypothesized to impair two aspects of locomotor performance (swimming and burying capacity). Peak swimming acceleration and both measures of burying performance did not differ between infected and uninfected fish, whereas swimming velocity of infected fish was significantly lower than that of uninfected fish. Smaller infected fish swam at significantly slower speeds than smaller uninfected fish, whereas there was no difference among larger fish. Neither the location nor the number of worms affected either swimming or burying performance. The decrease in swimming velocity observed in smaller infected fish may be sufficient in rendering them more vulnerable to predation and environmental stressors.