RAPD方法对中华绒螯蟹长江、辽河、瓯江三群体的遗传多样性分析

用RAPD技术检测了中华绒螯蟹（E．sinensis）长江、辽河、瓯江三个群体各10个个体的基因组DNA的多态性。经20个随机引物扩增得到每一个体的多态片段，通过对群体内及群体间共享片段的分析，得到三群体间的相似率数据，为中华绒螯蟹的种群关系的研究提供了重要的资料。     

江苏及安徽地区十个中华绒螯蟹成蟹群体的RAPD分析

采用随机扩增多态DNA(RAPD)技术对取自江苏及安徽地区的10个群体的中华绒螯蟹共64个个体进行了遗传多样性分析.从24个随机引物中筛选出15个扩增重复性好、条带清晰、特异性强的引物,共检测出88个位点(100～2000 bp),各群体的多态位点比例为3.45%～26.19%,遗传多态度为0.0141～0.1036,说明该蟹类基因组DNA多态度较为贫乏,遗传变异水平较低;各群体间的遗传距离为0.0237～0.2466,遗传相似度为0.7815～0.9766,该蟹类群体之间发生了一定程度的分化;分化系数为0.4283～0.6632,Nm为0.2539～0.6674,表明该蟹类群体之间发生了不同程度的遗传变异和遗传交换.     

巨野生群体遗传多样性的 RAPD 分析

巨（Bagarius yarrelli）为云南特有的经济鱼类，利用随机扩增多态性 DNA（RAPD）技术对元江下游的河口巨野生群体进行了遗传多样性分析，在50条随机引物中筛选出19条多态性较好的引物，通过 PCR 扩增，19条引物在巨群体中共检测到67个位点，其中多态性位点66个，平均多态性位点比率为98．51％，个体间平均遗传相似系数为0．8909，个体间平均遗传距离为0．1176；群体 Nei′s 基因多样性指数为0．2916， Shannon 信息指数为0．4269，结果表明巨野生群体具有较高的遗传多样性。     

中国大陆沿海六水系绒螯蟹（中华绒螯蟹和日本绒螯蟹）群体亲缘关系：R …

用４８个具有丰富多态性的１０碱基随机引物对辽河、黄河、长江、瓯江、珠江和南流江的中华绒螯蟹和日本绒螯蟹的６个群体进行ＲＡＰＤ分析。结果发现：（１）两个引物具有群体特异性带，其中Ｚ２扩增的８８０ｂｐ片段为珠江蟹和南流江蟹群体中所特有，而７００ｂｐ片段为长江蟹、辽河蟹、黄河蟹和瓯江蟹所特有，可作为区别日本绒螯蟹和中华绒螯蟹的分子遗传标记；（２）Ｏｐｐ１７扩增的９４７ｂｐ片段的出现频率，在长江、黄河和辽     

欧、美中华绒螯蟹源于中国长江水系中华绒螯蟹的遗传证据

用包括Z2和Opp17两个10碱基随机引物在内的10个引物,对来自荷兰斯科克莱（Skodely)、美国加州圣何塞（San Jose)的中华绒螯蟹群体与中国长江水系中华绒螯蟹群体进行RAPD遗传比较分析。结果发现：（1）中华绒螯蟹群体特有的Z2引物扩增的700bp标记带（Z2^700bp）,在荷兰与美国2个中华绒螯蟹群体中同时出现,而不出现日本绒螯蟹南流江种群中特有的880bp标记带（Z2^880bp）,表明欧洲、美国中华绒螯蟹与中国中华绒螯蟹为同种Eriocheir sinensis,而非日本绒螯蟹Eriocheir japonicus;（2）Opp17引物扩增的947bp片段在中国长江、荷兰及美国3个中华绒螯蟹群体内的出现频率均达100%。结合Z2引物扩增结果,欧洲与美国中华绒螯蟹群体极可能是从中国长江水系中华绒螯蟹引入繁衍的。     

中华绒螯蟹、日本绒螯蟹及其杂交F_1代群体的遗传多样性AFLP分析

通过4对引物组合对辽河水系中华绒螯蟹群体,俄罗斯的日本绒螯蟹群体及其杂交F1代(辽河蟹♀×俄罗斯蟹♂)的遗传多样性进行了AFLP分析。结果表明,4对引物共扩增出233条带(100bp~1000 bp),其中辽河群体多态条带184条,多态位点比例78.97%;俄罗斯群体177条多态带,多态位点比例75.97%;杂交群体182条多态带,多态位点比例78.11%。3个群体的群体内遗传相似度分别为0.7074±0.0469,0.7171±0.0475,0.7406±0.0449,香农多样性指数分别为0.185±0.128,0.194±0.119,0.176±0.138。变异来源有79.13%来自群体内,20.87%来自群体间。     

牙鲆和大菱鲆养殖群体的分子标记和遗传变异

用１６个１０碱基随机引物对养殖牙鲆（Ｐａｒａｌｉｃｈｔｈｙｓ ｏｌｉｖａｃｅｕｓ）和大菱鲆（Ａｃｏｐｈｔｈａｌｍｕｓ ｍａｘｉｍｕｓ）进行基因组随机扩增多态性ＤＮＡ（ＲＡＰＤ）遗传分析。发现：（１）多个引物具有种的特异性扩增带。其中引物Ｓ７扩增的６９０和１１６０ｂｐ带,引物Ｓ２扩增的６８０ｂｐ带为牙鲆的种的特异性谱带;而引物Ｓ７扩增的６５３和８８９ｂｐ带,引物Ｓ２扩增的６５０、８７０、９９０及１１     

中华绒螯蟹RAPD扩增条件及个体间遗传差异

该项研究优化了河蟹ＲＡＰＤ反应体系，初步确定：Ｔａｑ酶用量为ＩＵ，引物浓度０．３５μｍｏｌ／Ｌ，ｄＮＴＰｓ浓度１００μｍｏｌ／Ｌ模板量５０－１００ｎｇ，ＭｇＣｌ２浓度２．５ｍｍｏｌ／Ｌ，退火温度３８℃，得出同一养殖群中不同个体河蟹遗传差异在２０％以下。     

长吻遗传多样性的RAPD分析

以洞庭湖和原种场长吻（Leiocassis longirostris）为研究对象,采用20个随机引物对10个野生个体进行了随机扩增多态性DNA（random amplified polymorphic DNA,RAPD）群体遗传多样性分析。共检测到103条带,每个引物产生的条带数在2～9之间,片段大小在0.2～3.0kb之间,多态座位比例为41.75%,2群体内个体间遗传相似系数和遗传距离分别是,洞庭湖个体间遗传相似性系数（S）0.8795～0.9833,遗传距离（D）0.0167～0.1205;原种场个体间S为0.8794～0.9892,D为0.0319～0.1108;比较2群体S为0.7983～0.9994,D为0.0167～0.3017;Nei遗传多样性指数（He）0.3269。结果表明,长吻群体内遗传多样性较为丰富。     

不同鲤鱼种间DNA遗传标记多态性

用８４个随机引物对黑龙江野鲤（Ｃｙｐｒｉｎｕｓ ｃａｒｐｉｏ ｈａｅｍａｔｏｐｔｅｒｕｓ Ｔｅｍｍｉｎｃｋ ｅｔ Ｓｃｈｌｅｇｅｌ）和柏氏鲤（Ｃｙｐｒｉｎｕｓ ｐｅｌｌｅｇｒｉｎｉ ｐｅｌｌｅｇｒｉｎｉ Ｔｃｈａｎｇ）进行种间ＲＡＰＤ分析，结果共扩增出４８４个片段，二者共有扩增片段为１３个，黑龙江野鲤特有的为２３３个，柏氏鲤特有的为２２５个，这些特异片段可做为黑龙江野鲁和柏氏鲤的分子遗传标记。并对     

Protein and amino acid composition from the mantle of juvenile O ctopus vulgaris exposed to prolonged starvation

Protein and amino acid composition of the mantle of juvenile O ctopus vulgaris (Cuvier, 1797) during fasting for 27 days were determined. Average protein content of octopus mantle was of 711.19 ± 46.80 g kg^?1 DW, and it decreased with increasing fasting days. The non‐essential amino acids content was higher (486.18 ± 11.08 g kg^?1 protein) than essential amino acids (425.82 ± 9.15 g kg^?1 protein) at the start of the experiment (unstarved animals). The results suggest that the amino acid profile of the mantle where the most abundant amino acids are Arg, His, Lys, Gly, Leu and Pro could indicate a prolonged fasting condition (>20 days) or poor nutrition of O . vulgaris. This study supports the idea of using mantle for metabolic needs of starved O . vulgaris suggesting that the degradation pathway of amino acids to pyruvate and tricarboxylic acid cycle intermediates was favoured contrary to the degradation pathway of ketogenic amino acids. Special considerations should be taken concerning Thr, Ile, Ser, Ala, Asx (Asp, Asn), Glx (Glu, Gln) (because of their fast intake) and Lys and His (due to their stable contents) during a prolonged period of fasting.     

美人鱼发光杆菌对凡纳滨对虾非特异性 免疫功能及抗病力的影响

在基础饲料中分别添加0.1%和1%美人鱼发光杆菌灭活菌、0.1%美人鱼发光杆菌活菌配制成3种免疫实验饲料,以基础饲料为空白对照组饲料,每组设3个平行样。对个体质量为(4.83±0.36)g的凡纳滨对虾进行为期20 d的饲养实验,分别在0、5、10、15和20d进行取样,以血清中的酚氧化酶(PO)、酸性磷酸酶(ACP)、碱性磷酸酶(AKP)、超氧化物歧化酶(SOD)和溶菌酶(UL)活性为免疫指标,探讨了美人鱼发光杆菌作为免疫制剂对凡纳滨对虾非特异性免疫效应的影响;在投喂免疫饲料后的第22天,按0.004 2 kg/kg体重的剂量,直接投喂对虾白斑综合征病毒(WSSV)病料,并记录累积死亡率。结果表明,美人鱼发光杆菌免疫实验组对凡纳滨对虾血清中PO、ACP、AKP、UL和SOD活性影响明显高于对照组,并且在饲料中添加美人鱼发光杆菌后,明显提高了对虾抵御WSSV感染的能力。其中0.1%美人鱼发光杆菌活菌实验组的抗病毒感染能力最强,WSSV感染14d内累计死亡率为63.3%±5.8%;而对照组为96.7%±3.3%。研究表明,美人鱼发光杆菌添加在对虾饲料中能提高凡纳滨对虾非特异性免疫水平,增强抵抗疾病的能力,将其作为对虾免疫增强剂具有良好的应用前景。     

Effects of Spirulina platensis as a feed additive on growth and coloration of blue dolphin cichlids (Cyrtocara moorii Boulunger, 1902)

This study was carried out to investigate the effects of replacing fish meal with dietary Spirulina as a feed supplement on the growth performance and coloration of blue dolphin cichlids (Cyrtocara moorii). Five isonitrogenous (47% crude protein) and isocaloric (17.36 kJ/g digestible energy) diets were for formulated to replace FM with 0, 5, 10, 15 and 20% Spirulina (designated as Control, SP5, SP10, SP15 and SP20 respectively) and fed to the fish (initial body weight, 3.15 ± 0.01 g). Fish were randomly distributed into fifteen 120 L aquariums (26.5 ± 1.00°C), 15 fish per aquarium. The diets were tested in triplicate for 12 weeks. Experimental groups were fed twice daily (09:00 and 17:00) by hand to satiation. At the end of the feeding trial, significantly (p < 0.05) higher weight gain (WG), specific growth rate (SGR), protein efficiency ratio (PER) and lower feed conversion ratio (FCR) were observed in fish fed the SP10 diet when compared to the SP20 diet. There was no significant difference in these parameters between the other groups. The skin coloration of blue dolphin cichlid fed a diet containing Spirulina meal was enhanced. The best coloration was observed in the SP15 group. These impressions were objectively validated by chemical determinations of carotenoids extracted from fish skins and passed statistical tests of significance. The study findings show that Spirulina meal does not diminish growth rates except at very high levels.     

A Minchinia mercenariae‐like parasite infects cockles Cerastoderma edule in Galicia (NW Spain)

The cockle Cerastoderma edule fishery has traditionally been the most important shellfish species in terms of biomass in Galicia (NW Spain). In the course of a survey of the histopathological conditions affecting this species in the Ria of Arousa, a haplosporidan parasite that had not been observed in Galicia was detected in one of the most productive cockle beds of Galicia. Uni‐ and binucleate cells and multinucleate plasmodia were observed in the connective tissue mainly in the digestive area, gills and gonad. The parasite showed low prevalence, and it was not associated with abnormal cockle mortality. Molecular identification showed that this parasite was closely related to the haplosporidan Minchinia mercenariae that had been reported infecting hard clams Mercenaria mercenaria from the Atlantic coast of the United States. The molecular characterization of its SSU rDNA region allowed obtaining a fragment of 1,796 bp showing 98% homology with M. mercenariae parasite. Phylogenetic analysis supported this identification as this parasite was clustered in the same clade as M. mercenariae from the United States and other M. mercenariae‐like sequences from the UK, with bootstrap value of 99%. The occurrence of M. mercenariae‐like parasites infecting molluscs outside the United States is confirmed.     

Dietary thiamin and pyridoxine requirements of fingerling Indian major carp,Cirrhinus mrigala (Hamilton)

Two experiments were conducted to quantify the dietary thiamin (experiment I) and pyridoxine (experiment II) requirements of fingerling Cirrhinus mrigala for 16 weeks. In experiment I, dietary thiamin requirement was determined by feeding seven casein–gelatin‐based diets (400 g kg^?1 CP; 18.69 kJ g^?1 GE) with graded levels of thiamin (0, 0.5, 1, 2, 4, 8 and 16 mg kg^?1 diet) to triplicate groups of fish (6.15 ± 0.37 cm; 1.89 ± 0.12 g). Fish fed diet with 2 mg kg^?1 thiamin had highest specific growth rate (SGR), protein retention (PR), RNA/DNA ratio, haemoglobin (Hb), haematocrit (Hct), RBCs and best feed conversion ratio (FCR). However, highest liver thiamin concentration was recorded in fish fed 4 mg thiamin kg^?1 diet. Broken‐line analysis of SGR, PR and liver thiamin concentrations exhibited the thiamin requirement in the range of 1.79–3.34 mg kg^?1 diet (0.096–0.179 μg thiamin kJ^?1 gross energy). In experiment II, six casein–gelatin‐based diets (400 g kg^?1 CP; 18.69 kJ g^?1 GE) containing graded levels of pyridoxine (0, 2, 4, 6, 8 and 10 mg kg^?1 diet) were fed to triplicate groups of fish (6.35 ± 0.37 cm; 1.97 ± 0.12 g). Fish fed diet containing 6 mg kg^?1 pyridoxine showed best SGR, FCR, PR, RNA/DNA ratio, Hb, Hct and RBCs, whereas maximum liver pyridoxine concentration was recorded in fish fed 8 mg kg^?1 dietary pyridoxine. Broken‐line analysis of SGR, PR and liver pyridoxine concentrations reflected the pyridoxine requirement from 5.63 to 8.61 mg kg^?1 diet. Data generated during this study would be useful in formulating thiamin‐ and pyridoxine‐balanced feeds for the intensive culture of this fish.     

Sea bream culture in Egypt; status, constraints and potential

Marine fish farming in Egypt began in 1976 with the culture of gilthead sea bream, (Sparus aurata) as this fish was notably adaptable to brackish and marine pond conditions. Today, marine fish and shrimp farms amount to about 19,000 ha, out of which 42% is already in production while the rest, i.e., 58% is still under construction. In 1997, cultured gilthead sea bream production of 2,250 tons made up 3% of the 75,000 tons total aquaculture catch. In polyculture, usually with the grey mullet and sea bass, gilthead sea bream contributed 440 kg ha^–1 to the total yield of 1,700 kg ha^–1 (26%) over a period of 16 months. For the same period, the yield of monoculture ponds averaged 100 kg ha^–1, while in marine cages, yields ranged from 4–10 kg m³. In 1996–1997, fry of 0.25–1 g and fingerlings 1–10 g with a total of 3 million, were collected from the wild and 1 million fry were produced in the three marine hatcheries out of the four existing ones. The development of sea bream culture in Egypt is now severely inhibited by the shortage of seeds and adequate feeds. Exports of both sea bream and sea bass, during 1994–1996 averaged 1,300 tons per year.     

Fertilization by refrigerator stored sperm of the Indian major carp, Labeo calbasu (Hamilton, 1822)

This study reports on the spermatological properties, and on the development of a protocol for refrigerator storage (4°C) of Labeo calbasu (Hamilton, 1822) sperm for artificial breeding. Volume, motility, concentration and pH of the freshly collected sperm were 2.21 ± 0.53 (μL g^?1 of fish weight) (mean ± SD), 95 ± 1 (%), 1.93 ± 0.44 × 10⁹ (cells mL^?1) and 7.56 ± 0.17 respectively. Sperm activation was evaluated at different osmolalities of NaCl solution, and motility ceased completely when osmolality of the extender was ≥287 mOsmol kg^?1. Sperm retained motility for 24, 72 and 108 h, after refrigerator storage when sperm were undiluted, suspended in Alsever's solution and suspended in Alsever's solution containing 5% methanol respectively. Fertilization rate of fresh eggs with sperm stored at 4°C in Alsever's solution and Alsever's solution containing 5% methanol was 77% and 60% with a hatching rate of 60% and 43% respectively. The fertilization and hatching success of the stored sperm suggests potential to use refrigeration for transporting genetic material to hatcheries for artificial breeding of L. calbasu in Bangladesh.     

A novel approach to denitrification processes in a zero-discharge recirculating system for small-scale urban aquaculture

This paper presents an innovative process to solve the nitrate build-up problem in recirculating aquaculture systems (RAS). The novel aspects of the process lie in a denitrification bioreactor system that uses solid cotton wool as the primary carbon source and a unique degassing chamber. In the latter, the water is physically stripped of dissolved gaseous O₂ (by means of a Venturi vacuum tube), and the subsequent denitrification becomes more efficient due to elimination of the problems of oxygen inhibition of denitrification and aerobic consumption of cotton wool. The cotton wool medium also serves as a physical barrier that traps organic particles, which, in turn, act as an additional carbon source for denitrification. Operation in the proposed system gives an extremely low C/N ratio of 0.82 g of cotton wool/g of nitrate N, which contributes to a significant reduction of biofilter volume. The additional advantage of using solid cotton wool as the carbon source is that it does not release organic residuals into the liquid to be recycled. Operation of the system over a long period consistently produced effluents with low nitrate levels (below 10 mg N/l), and there was only a very small need to replace system water. The overall treatment scheme, also incorporating an aerobic nitrification biofilter and a granular filtration device, produced water of excellent quality, i.e., with near-zero levels of nitrite and ammonia, a sufficiently high pH for aquaculture, and low turbidity. The proposed system thus provides a solution for sustainable small-scale, urban aquaculture operation with a very high recovery of water (over 99%) and minimal waste disposal.     

Effects of metomidate anaesthesia or transfer to pur sea water on plasma parameters in ammonia-exposed Atlantic salmon (Salmo salar L) in sea water

Atlantic salmon (Salmo salar L) postsmolts weighing 150 ± 53 g were exposed to 14–15 mg l^–1 TA-N (total ammonia-N) in sea water in 1 m³ tanks for 24h. Blood samples were then taken A) immediately after the fish were netted from the exposure tanks and stunned by a blow to the head; B) 2–20 min after the fish were transferred to 15 l of an anaesthetic solution of metomidate in ammonia-free sea water; or C) 2–20 min after the fish were transferred to 15 l of ammonia-free sea water. Plasma TA-N level was 18% lower in the anaesthetised fish compared to in the fish sampled directly from the exposure tanks (p 0.05), and accordingly 16% lower in the fish transferred to pure sea water although this difference was not significant (p = 0.07). Plasma glucose level was higher in the fish transferred to pure sea water than in the fish receiving the two other treatments (p 0.05), but plasma urea, osmolality, Na^{+, Cl–, Ca2+ or Mg2+} levels did not vary significantly between the different treatments. Plasma TA-N level increased with time in the fish in the metomidate solution (p 0.02).