Using otolith weight to predict the age of <Emphasis Type="Italic">Pennahia macrocephalus</Emphasis> in the mouth of the Beibu Gulf

The relation between otolith weight (OW) and the age of marine fish is studied. A total of 222 individuals of bighead white croaker, Pennahia macrocephalus were sampled seasonally in the mouth of the Beibu Gulf, the South China Sea, in 2007. Since there are no significant differences in sagittal OW between otolith in pairs (P≥0.05), the undamaged left sagittal otolith is used for age determination. The highest correlations among standard length, OW and fish ages are confirmed by linear, exponential and multinomial regression. Results show that sagittal OW overlaps only occasionally among age groups, and to individuals with similar standard length, the older and slower-growing fish has a heavier otolith because of the continued otolith material deposition. There are differences in sagittal OW among different age groups and significant positive linear relationship with age (P<0.05). The age readings can be verified by plotting the sagittal OW versus the standard length for age groups, and the individuals with similar standard length but in different ages can be separated by sagittal OW frequency analysis. Mostly, the predicted ages using the regression between sagittal OW and ages are closed to the observed ages by counting annulus on scale. It indicates that the sagittal OW analysis is a useful technique for validating the accuracy of age determination by annuli counts, especially for individuals of similar size. Furthermore, the technique is applied for Pennahia macrocephalus with discussion in this paper. Supported by the National Natural Science Foundation of China (No. 30771653) and Bureau of Fisheries, Ministry of Agriculture     

Formation and early development of tetraspores of Polysiphonia urceolata （Rhodomelaceae, Rhodophyta）

Polysiphonia urceolata is one type of potential commercial red seaweeds used for breeding and cultivation, because of its significant biochemical and biomedical application. However, the information of breeding and seedling incubation for cultivation is limited, especially the early development. In this study, tetrasporohyte and gametophyte of P. urceolata were taken as the study materials in Huiquan Bay, Qingdao, China. The cleaned and sterilized tetrasporophytes and gametophytes were pre-cultured in sterilized seawater, then nurtured at 18°C, 25 μmol photons m⁻² s⁻¹ in 12:12 h (light:dark) photoperiod. Continuous observation under microscope showed that the early development consists of bipolar division stage and seedling stage. In the division stage, tetraspores germinate into bipolar sporelings that further differentiate into a colorless rhizoidal portion and a lightly pigmented upright shoot. The lightly pigmented rhizoidal cell develops to a rhizoid and the larger pigmented cell transforms to an erect axis. In the seedling stage, several quasi-protuberances appear on the erect axis and form juvenile seedlings. The results demonstrate the culture of P. urceolata from tetraspores under laboratory conditions. Supported by National Key Technology Support Program, Development Program of China (No.2006AA09Z21), National Natural Science Foundation of China (No. 40618001 and N_CUHK438/06) and Shandong Agricultural Seed Stock Breeding Project     

Initiation of primary cell culture from amphioxus <Emphasis Type="Italic">Branchiostoma belcheri tsingtauense</Emphasis>

Amphioxus, a cephalochordate, is an important model fish for studies in evolution and comparative biology. A successful cell culture from amphioxus tissues in vitro would help understanding some basic issues. To determine the optimal culture conditions for proliferation of amphioxus cells, primary cultures were initiated from buccal cirri, tail, gill, gut and metapleural fold of amphioxus Branchiostoma belcheri tsingtauense. The media tested were L-15, F-12, M 199, MEM, DMEM, PRMI 1640 and LDF, each was supplemented with 20% fetal bovine serum. The optimal conditions include tail tissue cultured in L-15 or F-12 with supplement of 20% FBS and 1.5% NaCl at about 25°C. Supported by Doctoral Initial Fund of Ludong University (No.43304)     

钻井柱状图可视化查询系统的设计与实现

钻井柱状图是石油行业勘探开发最常用的一种图件。为了开发出灵活通用的钻井柱状图制作软件,为可视化分析提供良好基础,提出了“两库四类三层”系统设计方案,应用COM思想和模板定制技术,采用C#．net语言从底层进行了软件实现,所开发的软件模块具备一定的独立性和较好的通用性。     

Construction of bacterial artificial chromosome libraries for Zhikong Scallop <Emphasis Type="Italic">Chlamys farreri</Emphasis>

Two Large-insert genomic bacterial artificial chromosome （BAC） libraries of Zhikong scallop Chlamys farreri were constructed to promote our genetic and genomic research. High-quality megabase-sized DNA was isolated from the adductor muscle of the scallop and partially digested by BamH I and Mbo I, respectively. The BamH I library consisted of 53 760 clones while the Mbo I library consisted of 7 680clones. Approximately 96 % of the clones in BamH I library contained nuclear DNA inserts in average size of 100 kb, providing a coverage of 5.3 haploid genome equivalents. Similarly, the Mbo I library with an average insert of 145 kb and no insert-empty clones, thus providing a genome coverage of 1.1 haploid genome equivalents.     

Effects of diet and body size on phosphorus utilization of Liza haematocheila T. ＆ S.

A 21-d laboratory experiment was conducted to study, the phosphorus （P） utilization of two different diets by redlip mullet Liza haematocheila T. ＆ S. Sand-filtered water in salinity 30 and temperatare 25℃ was used. Twenty-nine fish individuals were divided into three groups： 11 to group 1 （G1） fed on diet 1, 11 to group 2 （G2） fed on diet 2, and 7 to contrast group. Diet 1 was a commercial feed, more valuable in nutrition than diet 2 that similar to natural detritus. The results show the intake phosphorus （IP） of G1 was significantly higher than that of G2, and both increased linearly with body size at a certain amount of diet. The retention phosphorus （RP） in fish of G1 was lower than G2. The relationship between retention phosphorus and body size was positive and stronger in G2. Significant difference in faecal phosphorus （FP） was found between G1 and G2. Body size significantly impacted the excretion phosphorus （EP） in G1 but G2. The loss of intake phosphorus in G1 was 10.83-20.27 mg per g fish weight gain, higher than that in G2 for 6.63-9.56. Of the phosphorus, about 10% was allocated into growth, 50% in faeces, and the rest lost in excretion. The main part of phosphorus was lost in faeces but excretion. The phosphorus budget of the fish could be described as 100IP = 7.40RP ＋ 47.39FP ＋ 36.63EP （Diet 1） or 100IP = 11.93RP ＋ 56.64FP ＋ 21.76EP （Diet 2）.     

The hydrogen peroxide impact on larval settlement and metamorphosis of abalone <Emphasis Type="Italic">Haliotis diversicolor supertexta</Emphasis>

Abalone Haliotis diversicolor supertexta is an important economic mollusk. The settlement and metamorphosis are two critical stages during its development period, which has direct influence on abalone survival and production. The influence of reactive oxygen species （hydrogen peroxide） on abalone embryo and juvenile development were examined in this study. Larvae of Haliotis diversicolor supertexta were induced to settlement and metamorphose by exposure to seawater supplemented with hydrogen peroxide. They had the best performance at 800 μmol/L. The concentration of 1 000 μmol/L or higher was toxic to the larvae, as the larvae could settle down only at benthic diatom plates without complete metamorphosis. In addition, H2O2 adding time was critical to the larval performance. 24h after two-day post-fertilization was proved to be the optimal adding time. In this paper, two action mechanisms of hydrogen peroxide are discussed： （1） hydrogen peroxide has direct toxicity to ciliated cells, thus cause apoptosis; （2） hydrogen peroxide, as a product from catecholamines＇ autoxidation process in vivo, can reverse this process to produce neuro-transmitters to induce abalone metamorphosis.     

Molecular phylogenetic analysis of attached Ulvaceae species and free-floating <Emphasis Type="Italic">Enteromorpha</Emphasis> from Qingdao coasts in 2007

Based on the sequence data of the nuclear ribosomal DNA internal transcribed spacer (ITS) 1, 5.8 S, and ITS 2, the molecular phylogeny was analyzed on Ulvaceae species collected from Qingdao coasts in summer of 2007, including 15 attached Ulva and Enteromorpha samples from 10 locations and 10 free-floating Enteromorpha samples from seven locations. The result supported the monophyly of all free-floating Enteromorpha samples, implying the unialgal composition of the free-floating Enteromorpha, and the attached Ulvaceae species from Qingdao coasts were grouped into other five clades, suggesting that they were not the biogeographic origin of the free-floating Enteromorpha in that season. Supported by NSFC (40506030); the Innovative Key Project of the Chinese Academy of Sciences (KZCX2-YW-209); Science & Technology Project of Qingdao City (06-2-2-12-JCH)     

Non-specific immune response of bullfrog <Emphasis Type="Italic">Rana catesbeiana</Emphasis> to intraperitoneal injection of bacterium <Emphasis Type="Italic">Aeromonas hydrophila</Emphasis>

Non-specific immune response of bullfrog Rana catesbeiana to pathogenic Aeromonas hydrophila was studied to 60 individuals in two groups. Each bullfrog in bacterium-injected group was injected intraperitoneally （i.p.） with 0.2 ml bacterial suspension at a density of 5.2 × 10^6 CFU/ml, while each one in control group injected i.p. with 0.2 ml sterile saline solution （0.85%, w/v）. Three bullfrogs in both groups were sampled at 0, 1, 3, 7, 11, 15 and 20 days post-injection （dpi） for the evaluation of non-specific immune parameters. It was observed that intraperitoneal injection of A. hydrophila significantly increased the number of leucocytes and that of NBT-positive cells in peripheral blood. Significant increases in serum bactericidal activity and serum acid phosphatase activity were also observed in the bacterium-injected frogs when compared with those in the control group. However, a significant reduction was detected in vitro in phagocytosis activity of peripheral blood phagocytes. No significant difference in changes in the number of peripheral erythrocytes, serum superoxide dismutase （SOD） activity, and lysozyme activity was detected between the two groups. It is suggested that bullfrogs may produce a series of non-specific immune reactions in response to the A. hydrophila infection.