西伯利亚鲟不同性别与卵巢发育时期血液性类固醇激素的差异与判别分析

利用电化学发光免疫分析法(ECLIA)测定了人工养殖西伯利亚鲟(Acipenser baerii baerii)不同性别和卵巢发育时期血浆中睾酮(T)和雌二醇(E2)的含量。利用活组织取样的方法,根据性别和卵子颜色及大小将实验鱼分为成熟雄鱼(M)、雌鱼Ⅲ期、Ⅳ期和Ⅴ期共4个组。研究结果表明,西伯利亚鲟血浆T含量雄鱼高于雌鱼(P<0.01),且雄鱼与雌鱼Ⅲ期和Ⅴ期组有极显著性差异(P<0.01),而E2含量除与雌鱼Ⅳ期有极显著性差异外(P<0.01),与雌鱼Ⅲ期和Ⅴ期组无显著性差异(P>0.05),雄鱼表现为T含量和T/E2比值高,而E2含量低。雌鱼随卵巢的发育T和E2含量均表现为"先上升,后下降"的趋势,即Ⅳ期组性类固醇激素的含量明显高于Ⅲ期和Ⅴ期,且差异极显著(P<0.01)。根据血浆中T和E2的含量建立了西伯利亚鲟性别和性腺发育时期的4个判别函数:YM=-11.322+0.043T+0.153E2,YⅢ=-1.390+0.000T+0.061E2,YⅣ=-18.074+0.032T+3.305E2,YⅤ=-2.316+0.785T+0.008E2,经检验总体判别准确率为95%,其中对性别的判别准确率为100%,对雌鱼卵巢发育时期的判别准确率为93%。     

西伯利亚鲟性别及性腺发育的超声波鉴定

利用超声波检查仪对人工养殖西伯利亚鲟的性别及性腺发育进行超声波扫描,并利用微创手术检查和组织学观察法来验证超声波对西伯利亚鲟性别及发育时期扫描的鉴定结果。通过对比研究表明:超声波方法对西伯利亚鲟性别鉴定的准确率较高,对2～5龄雌性鉴定的准确率为95%,对雄性鉴定的准确率为87%。超声波技术鉴定性腺发育的准确率随性腺的发育而提高,鉴定早期性腺(Ⅰ～Ⅱ期)发育的准确率(65.25%)低于对晚期性腺(Ⅲ～Ⅴ期)发育鉴定的准确率(88.65%)。超声波技术鉴定西伯利亚鲟性别及性腺发育速度较快,根据鱼的发育状况,平均每尾鱼所用的时间约30s。     

福氏刺尻鱼性逆转的配对诱导及组织学观察

本研究通过人工配对方式对福氏刺尻鱼的性逆转进行了诱导，并从形态学和组织学层面对其性逆转过程和性腺发育进行了研究。结果表明，福氏刺尻鱼的性腺分为纯雌鱼卵巢、纯雄鱼精巢和雌雄（♀/♂）兼性期性腺3种类型。性腺均先分化为卵巢；性逆转发生于第II期卵巢阶段，通过1大1小配对诱导1~2周时，大个体雌鱼第II期卵巢停止发育并逐渐萎缩，在卵巢壁和产卵板上逐渐分化出雄性生殖细胞，性腺内雌雄生殖细胞共存，形成兼性期性腺，即为雌雄同体阶段；配对诱导3周后，兼性期性腺完全分化为早期精巢，完成性逆转，个体成为功能性雄鱼；配对诱导4周后，性逆转的雄鱼精巢和小个体雌鱼的卵巢发育成熟，开始排精产卵。本文还分析了福氏刺尻体长、体重与性逆转的关系，并对性逆转过程、特点及可能的诱导机制进行了探讨。研究结果可为开展福氏刺尻鱼人工繁育和性别控制技术研究提供理论指导，也为揭示其性腺发育及性逆转机制奠定了基础。     

仿洄游培育美洲鲥亲本的性腺发育

为解决美洲鲥(Alosa sapidissima)人工培育亲本性腺发育不佳的问题,开展了美洲鲥亲本仿洄游的培育,研究观察了美洲鲥亲本卵巢和精巢不同发育阶段(II^V期)的形态特征及相关参数,确定了仿洄游培育美洲鲥亲本性腺初次发育的特征。结果显示:在美洲鲥仿洄游培育过程中,在卵巢、精巢发育前期(II期),雌雄亲本生长迅速;而性腺发育到III期后,雌雄亲本的生长滞缓,雌鱼亲本甚至出现负生长。仿洄游培育美洲鲥性腺初次发育(II^V期)过程中,雌雄亲本均有较高的肥满度,分别为1.55~2.07和1.56~1.71;卵巢发育Ⅴ期雌鱼亲本的肥满度(2.07)显著高于在卵巢II^IV期亲本(1.55~1.67);而雄鱼亲本肥满度在精巢发育各阶段(II^V期)均无显著性差异。仿洄游培育美洲鲥性腺发育良好,雌雄亲本的性腺指数均较高,性腺发育Ⅴ期雌雄亲本的性腺指数(GSI)分别为(26.10±6.46)(18.88~33.62)和(5.18±0.68)(4.49~6.11)。研究表明,仿洄游培育的水温(10.8~30.8℃)和盐度(1~15)的周年变化符合美洲鲥亲本性腺发育的要求。     

西伯利亚鲟的人工繁殖

取7龄西伯利亚鲟(AcipenserbaeriBrandt),第1次产卵前4个月,放入2组不同温度的水池中越冬,Ⅰ组水温13～14℃;Ⅱ组7～8℃。每组4尾雌鱼,6尾雄鱼。催产前水温升至15～16℃,人工催产结果,Ⅰ组1尾雌鱼排卵和2尾雄鱼排精,Ⅱ组3尾雌鱼排卵,5尾雄鱼排精;雌鱼排卵全部采用输卵管切割手术取卵,获得成熟卵3.26kg,共计21.5×104粒;用“半干法”授精,获受精卵19.11×104粒,受精率86.7%～95.6%;孵出鱼苗12.5×104尾,孵化率40%～68.9%。结果说明,在人工培育条件下,西伯利亚鲟能够成熟并成功地进行人工繁殖;产前培育温度变化对西伯利亚鲟性腺发育和人工繁殖效果有明显影响;与养殖施氏鲟的人工繁殖比较,催产效应时间基本相同;雄鱼发育较差。     

多鳞铲颌鱼性腺分化发育的组织学研究

用光镜和透射电镜对多鳞铲颌鱼（Varicorhinus macrolepis）性腺的分化发育进行组织学研究，观察了雌雄性腺的形成过程及各级生殖细胞的形态特征。结果表明，多鳞铲颌鱼卵巢和精巢开始分化的时间分别在80日龄和90日龄，雌鱼和雄鱼的性成熟年龄分别为36月龄和24月龄，精子和卵子的发育均可分为5个时期。在多鳞铲颌鱼的生长发育过程中，12月龄（雄鱼）和30月龄（雌鱼）性腺质量与体长、体质量之间具有极显著的相关关系。     

西伯利亚鲟卵巢发育过程血清脂蛋白脂酶活性与血脂代谢的研究

利用微创手术检查的方法对人工养殖西伯利亚鲟的卵巢发育期进行鉴别,根据卵巢发育期将西伯利亚鲟分为Ⅱ期、Ⅲ期、Ⅳ期、Ⅴ期和Ⅵ期共5个实验组。对5个实验组血清脂蛋白脂酶(LPL)活性及血脂含量进行测定,并对其之间的关系进行分析。结果表明:随着卵巢发育,血清LPL活性、极低密度脂蛋白胆固醇(VLDL-C)、甘油三酯(TG)呈"先上升,后下降"的趋势,其中LPL在Ⅴ期达到最高值,VLDL-C、TG均在Ⅳ期达到最高值;总胆固醇(TC)、低密度脂蛋白胆固醇(LDL-C)随卵巢发育表现为上升趋势。血清TG与LPL呈正相关关系。     

幼鱼阶段电刺激对稀有鮈鲫性腺发育及繁殖的影响

控制水域电压为75 V,电流280 m A,持续电刺激5 500 ms左右,电刺激的作用强度是使鱼既能达到休克昏迷状态又能在几分钟内苏醒。通过对1月龄的稀有鮈鲫(Gobiocypris rarus)进行电刺激,观察电刺激对其性腺发育及性成熟后繁殖的影响,以期为渔政部门评价电捕鱼行为对鱼类资源的影响提供参考。结果表明,电刺激1次组、电刺激3次组和电刺激5次组可以促进雌雄鱼的生长和性腺发育,连续电刺激组对雄鱼的生长和性腺发育影响不大却会抑制雌鱼的生长和性腺发育;电刺激1次组、电刺激3次组和电刺激5次组对性成熟后雌雄鱼的繁殖没有产生显著差异,而连续电刺激组对性成熟后雄鱼的繁殖没有产生显著差异,却会造成雌鱼不产卵、或产出的卵颜色泛白,且可产卵雌鱼的产卵周期平均延长4.5 d,产卵量平均减少36粒,受精率平均降低9.2%。组织切片观察发现,电刺激1次组、电刺激3次组和电刺激5次组对性成熟后雌鱼的卵巢没有造成影响,而连续的周期性电刺激对性成熟后雌鱼卵巢中大的卵细胞的细胞膜造成了损坏,细胞膜界限变得不清晰;电刺激对性成熟后雄鱼精巢没有造成影响。     

养殖银鲳性腺发育规律和性类固醇激素变化

为掌握养殖银鲳的性腺发育规律及其与性类固醇激素水平的关系,本研究采用组织切片技术和酶联免疫吸附法(Elisa),观察养殖条件下银鲳的精巢与卵巢发育特征和血清中性类固醇激素的周年变化,并分析性腺成熟指数(GSI)、肝重指数(HSI)和肥满度(CF)与性腺发育的关系。结果显示,养殖银鲳雌鱼1龄即可发育成熟,成熟卵巢呈一对""形的囊状结构。11月份有近一半个体发育至III期,33.3%已发育至IV期,12月—翌年1月越冬期间约2/3退化至II期,3月又迅速发育至IV期,4月份达到V期并产卵,5月份产卵结束卵巢进入恢复期IV期。银鲳精巢属小叶型,III期精巢出现小叶腔,IV期精巢出现精子,精巢发育速率快于卵巢,即"雄性早熟",7月龄约20%个体可达到性成熟,越冬时退化至III期,3月重新发育,精子分批成熟排精。雌鱼GSI值为0.19%~12.89%,HSI为0.97%~2.30%,CF为2.30~3.08 g/cm~3,雄鱼的GSI为0.08%~2.62%,HSI值为0.73%~1.83%,CF为2.11~2.80 g/cm~3,GSI值在V期时达到最大,HSI和CF值则于IV期达到峰值。雌鱼血清中雌二醇(E2)周年表达水平为23.27~59.13 pg/m L,雄鱼为15.90~36.20 pg/m L,雌鱼的睾酮(T)为14.57~68.67 nmol/L,雄鱼的T为18.62~66.49 nmol/L,E2水平与HSI值呈现显著正相关关系,T表达水平与GSI值呈现显著正相关关系。研究表明,银鲳血清中性类固醇激素含量与精巢和卵巢的发育密切相关,可作为了解养殖银鲳性腺发育的重要指标。     

西伯利亚鲟性腺早期发生、分化、发育的组织学观察

利用组织学方法研究人工繁育、养殖的西伯利亚鲟(Acipenser baerii)仔、幼、成鱼性腺发育过程中原始生殖细胞(Primordial germ cells,PGC)的迁移,生殖褶的生成,性腺雌雄分化和发育过程。结果表明,西伯利亚鲟于3dph(Days post-hatch)时,PGC以单细胞的形式存在于仔鱼卵黄囊上方两中肾管之间,10dph时其迁移到背侧的腹膜上皮上,13dph时其沿腹膜上皮朝肾管区下方迁移,22dph时生殖嵴形成,31dph时原始性腺形成,40～60dph时原始性腺发育形成裂腔,PGCs沿腔壁排列,裂腔经过进一步发育并从背离腹膜上皮的底端慢慢愈合成实体,90dph时原始性腺中出现生殖上皮和性细胞,150dph时脂肪细胞和血管增多,210dph时性腺出现2种不同的表面结构,性腺开始分化。本研究利用组织学方法借鉴鱼类性腺分期标准,将西伯利亚鲟性腺分化划分为0期和Ⅰ～Ⅵ期,并描述了各期精巢、卵巢的结构变化以及细胞形态学特征。     

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.     

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.     

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.     

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).