单环刺螠消化道的发生和分化

用组织学、组织化学和电镜技术对单环刺螠（Urechis unicinctus）消化道的发生及分化进行研究。结果表明，单环刺螠消化道起源于原肠胚期内陷而成的原肠腔；至早期担轮幼虫时期，消化道连通，开始摄食，此时据管腔直径不同消化道可分为口、食道、胃、中肠及后肠。附着变态后，其消化道在结构及功能上都有了明显的发育与分化。至蠕虫状幼虫时期。食道、胃及后肠已出现黏膜下层及肌层。嗉囊与砂囊可明显区分。中肠已产生肌纤维的分化，呼吸肠由中肠分化而来，其胞体细长，且核质比较小，可与中肠明显区分。至幼螠时期，食道前端产生了咽部的分化，至此消化道各部位在结构及功能上都与成体有了很大的相似性。即由口、咽、食道、嗉囊、砂囊、胃、中肠、呼吸肠、后肠和肛门组成。组织化学研究结果表明，随着发育，消化道各部位分别呈现不同的碱性磷酸酶、酸性磷酸酶、脂酶、ATP酶、非特异性酯酶及多糖类物质活性。     

单环刺螠Vasa基因的早期发育表达图式

Vasa基因编码DEAD-box家族成员中一种ATP依赖的RNA解旋酶,是决定生殖系发育的重要调控因子之一。采用同源克隆策略及SMART-RACE技术,克隆了海洋经济螠虫动物单环刺螠Vasa基因的全长cDNA,该序列长4 080 bp,开放阅读框2 322 bp,编码733个氨基酸,具有DEAD-box蛋白家族共有的全部9个保守域。整体原位杂交和半定量RT-PCR结果显示,Vasa基因在未受精卵、受精卵、2～8细胞的早期胚胎中均有明显的表达,显示其母源性提供的特点;从囊胚开始,表达量明显降低,在原肠胚表达信号主要集中在内中胚层细胞中;至担轮幼虫,表达信号进一步集中在消化道处;当发育至体节幼虫时,阳性细胞分布于消化道和体节的隔膜处,进入蠕虫状幼虫,信号仅在头部的腹刚毛附近以及后肠周围的细胞中表达。实验结果为探知刺螠动物生殖系的起源和分化以及低等型生殖腺的发生提供重要的数据。     

单环刺螠的繁殖生物学

运用实验生态学和组织学方法研究了单环刺螠的性比、性腺指数、繁殖力、胚胎与胚后发育等繁殖生物学特征,并采用正交实验法分析了单环刺螠受精率和孵化率的最佳环境条件,为单环刺螠的人工增养殖提供科学依据。结果显示,单环刺螠雌、雄比例为1∶1.17,雄性略多于雌性。成熟卵径为(145.16±1.70)μm×(141.27±1.94)μm,处于第1次成熟分裂前期。绝对繁殖力为111 800～720 488(449 684±265 905)粒,相对繁殖力为2 214～12 702(6 849±4 854)粒,怀卵量与体质量呈显著正相关。生物学最小型体长约7 cm,体质量为11.21 g。精子密度为(5.88±0.52)×109个/mL,雄性个体平均精子数为(2.41±0.73)×1010个/只。雌性性腺指数在繁殖季节4—5月份呈现2个峰值,表明单环刺螠为分批产卵类型。在盐度25、水温15°C的条件下,胚胎发育经20～24 h孵化。组织学研究表明,单环刺螠为间黄卵,进行不等全裂,且有螺旋卵裂的特征,形成偏极囊胚,原肠作用方式为外包和内陷。胚后发育经担轮幼虫(孵化后1～10 d)、体节幼虫(11～20 d)、蠕虫状幼虫(21～30 d)发育为幼螠。正交实验结果表明,单环刺螠受精率的最佳环境条件为温度25°C、盐度35、pH 8～9;孵化率的最佳环境条件为温度15°C、盐度25、pH 8～9。     

鄱阳湖日本沼虾幼体发育及其同工酶研究

对鄱阳湖日本沼虾(Macrobrachiumnipponensis)幼体发育的5个阶段即Z0、Z1、Z3、Z5～Z6及Z9～Z11期进行观察,分析乳酸脱氢酶(LDH)、酯酶(EST)、苹果酸脱氢酶(MDH)同工酶谱带变化。结果表明,各期间幼体形态均有明显变化,幼体蜕皮次数随发育快慢不同,发育快的需9次蜕皮完成变态发育过程,慢的则需14次;LDH酶谱在5个发育阶段各有5条带,仅酶活性有强弱变化;EST酶谱不仅条带数从2条增加到9条,酶活性也有明显变化,EST1和EST3在5个阶段都出现,且酶活性逐渐升高;MDH酶谱共出现6条带,MDH1和MDH4分布延伸时间跨度大,酶活性高,可看作主带。因此日本沼虾变态发育过程中同工酶谱带随变态发育而变化,反映了不同组织器官细胞组成和不同组织中不同同工酶的活性和出现顺序,可作为鉴别种属特异性和发育阶段特异性的一项生化指标。     

罗氏沼虾胚胎发育期主要消化酶和同工酶特性

采用生物化学方法分别测定了罗氏沼虾胚胎发育期5种消化酶:胃蛋白酶、类胰蛋白酶、淀粉酶、纤维素酶和脂肪酶的比活力,测定了6种同工酶:酯酶(EST)、苹果酸脱氢酶(MDH)、谷氨酸脱氢酶(GDH)、碱性磷酸酶(ALP)、乳酸脱氢酶(LDH)和淀粉酶(AMY)在成熟卵细胞和胚胎发育时期的表达特征。结果显示:5种消化酶的活力表现出两种变化趋势,即胃蛋白酶、类胰蛋白酶、淀粉酶和纤维素酶的酶活力呈现出“高—低—高”的变化趋势,而脂肪酶活力则呈现出下降的变化趋势。6种同工酶酶谱则随胚胎发育渐趋复杂,酶活性随之增强。研究表明,所测定的消化酶以及同工酶在罗氏沼虾受精卵期均已表现出酶活性,此阶段为卵源性的酶;原肠期是胚胎发育中一个较为重要的时期,胚胎内酶基因表达能力开始逐渐增强,开始合成一些酶类,为胚胎发育后期利用卵黄物质提供了保证。消化酶以及同工酶对卵黄物质的利用和胚胎发育的物质代谢起重要作用。     

点带石斑鱼与褐点石斑鱼4种同工酶的比较研究

采用聚丙烯酰胺凝胶垂直梯度凝胶电泳法,对点带石斑鱼与褐点石斑鱼7种组织中的4种同工酶(EST、LDH、MDH、SOD)的组织特异性进行比较研究。试验结果表明:2种石斑鱼的4种同工酶均具有明显的组织特异性。2种石斑鱼同一组织间MDH酶谱没有差别;LDH酶谱相似,只在迁移率上略有差异;EST、SOD酶谱间存在不同程度的种间差异,可作为鉴定点带石斑鱼与褐点石斑鱼的遗传标记。     

单环刺螠浮游幼体饥饿实验及不可逆生长点的确定

正单环刺螠俗称海肠,是一种经济价值较高的海产品种。本文通过研究饥饿条件对单环刺螠浮游幼体生长和存活的影响,初步确定了单环刺螠浮游幼体最佳投饵时间及不可逆点,为单环刺螠人工育苗提供理论依据。单环刺螠(Urechis unicinctus)隶属螠虫动物门、螠纲、无管螠目、刺螠科、刺螠属。生活在海边潮间带,     

大鳞鲃鱼同工酶的研究

本实验采用聚丙烯酰胺垂直平板电泳方法,分析了大鳞鲃鱼的肝、肾、眼3种组织中的醋酶（EST）、淀粉酶（AMY）、乳脱酸氢酶（LDH）和苹果酸脱氢酶（MDH）4种同工酶。结果表明各种同工酶具有明显的组织特异性,肝脏酶活性最高,具有所有酶带。     

大鳞鲃鱼同工酶的研究

本实验采用聚丙烯酰胺垂直平板电泳方法,分析了大鳞鲃鱼的肝、肾、眼3种组织中的醋酶（EST）、淀粉酶（AMY）、乳脱酸氢酶（LDH）和苹果酸脱氢酶（MDH）4种同工酶。结果表明各种同工酶具有明显的组织特异性,肝脏酶活性最高,具有所有酶带。     

水温、盐度对单环刺螠幼体发育影响

<正>单环刺螠(Urechis unicinctus),俗称"海肠",是辽宁地区著名的土著水产经济品种,属螠虫动物门,螠纲,无管螠目,刺螠科,为真体腔原口动物。味道鲜美,富含蛋白质、人体必需氨基酸、多肽、糖胺聚糖等多种活性物质。近年来,过度捕捞和环境污染严重破坏了单环刺螠的自然资源,导致其采捕量急剧下降。随着人工育苗关键技术的突破[1-2],单环刺螠的人工养殖势在必行。该试验研究了水温、盐度对单环刺螠幼体发育的影响,从发育速度、成活率两个角度初步建立了适     

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.     

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.     

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.     

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.     

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.     

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).