暴发性鱼病菌苗的灭活条件及免疫效果的研究

０．０５％－１．０％的福尔马林、０．０１％－０．１％的新洁尔灭、３．０％－４．０％的氯仿以及６３℃高温均对嗜水气单胞菌灭活作用。温度对嗜水气单胞菌的灭活无明显影响，但低温灭活有利于菌苗抗原性的保持，受试鱼产生的保护作用更强，故以低温４℃时用３．０％－４．０％的氯仿、０．１％－０．２％的福尔马林，０．０３％－０．０５％的新洁尔灭制备的菌苗免疫效果最好。     

暴发性鱼病菌苗产生免疫力的时间及免疫时效研究

嗜水气单胞菌苗免疫接种试验表明，室温３０℃时，试验鱼注射接种１．８～２．７×１０８个／尾的灭活菌苗，８－１０天后即产生较强的免疫力，注射接种１％一１‰的灭活组织苗０．２－０．３ｍｌ／尾，６－８天后即产生较强的免疫力，两者的保护力均达９２％以上，浸泡相当于１．８×１０６个／ｍｌ的脂多糖粗提物（ＬＰＳ），１０—１２天后产生的保护力可达７０％左右。不同温度的接种试验表明，水温３０℃时接种比１５℃时接种，免疫力的形成早４—７天。大池试验表明，灭活苗苗接种鱼的免疫时效达６个月以上。     

团头鲂暴发性败血症病原分离鉴定及免疫防治

从患有暴发性败血症的团头鲂体内分离到两个菌株，经攻毒试验证明是致病菌。经16SrDNA序列分析鉴定为嗜水气单胞菌。采用两种不同条件对其进行灭活，分别为0．2％福尔马林25℃灭活24h和0．5％福尔马林4℃灭活24h，注射免疫团头鲂。通过测定受免团头鲂血清中抗体凝集效价和血液中吞噬细胞活性以评估免疫效果，结果显示两种方法制备的灭活菌苗均可刺激鱼体产生较强的免疫应答，且采用0．2％福尔马林灭活的免疫效果优于0．5％福尔马林灭活，但二者无显著差异。     

白鲢暴发病菌苗的制备及免疫效果分析

在３２℃下对嗜水气单胸菌Ｈ２０株作振荡培养，１％福尔马林、６０℃灭活１ｈｒ．，制备白鲢暴发病菌苗。用该菌苗免疫白鲢，１８－２０℃水温下饲养１０天后，白鲢血清中凝集抗体效价达１：１１２８左右，未免疫对照组凝集抗体效价约为１：８。免疫鱼的抗感染保护力达８０％。     

红螯螯虾对硫酸铜和孔雀石绿耐受性的研究

测定了红螯螯虾虾苗对硫酸铜和孔雀石绿的耐受性，结果表明，２４ｈＬＣ５０分别为６３．８２×１０＾－４％和１３．７８×１０＾－４％；４８ｈＬＣ５０分别为２３．３７×１０＾－４％和４．８４×１０＾－４％；９６ｈＬＣ５０分别为１５．０７×１０＾－４％和２．６５×１０＾－４％；安全浓度分别为０．９４×１０＾－４％和０．１８×１０＾－４％。     

中华鳖腐皮病的免疫预防研究

选择对中华鳖毒力最强的Ｐ－１菌株,经扩大培养制成灭活福尔马林菌苗（Ｆ－ＡＨ）及Ｓｐａｎ白油佐剂菌苗（Ｚ－ＡＨ）,对中华鳖进行了Ｆ－ＡＨ菌苗的１次注射、２次注射及Ｚ－ＡＨ菌苗的注射免疫试验。结果表明,受免疫组鳖血液白细胞吞噬活性、凝集抗体效价及脾脏吞噬活性均显著高于对照组鳖,免疫保护力均达到８８％以上。     

淡水虾苗种高温期运输技术探讨

利用增设攀缘设施，进行帆布袋充气运输的方法，保持充气量４４．８Ｌ／ｍ３·分钟。①８月份，水温２７－２９℃，虾苗规格４３００－４５００尾／公斤，起运密度以７．２－７．８万尾／ｍ３为好，这种情况下，运输９，５小时，成活率为９４．９％；运输１７．０－２１．５小时，成活率８７．３％。②９月份，水温２５－２８℃，虾苗规格２０００－２４００尾／公斤，起运密度以４．３－４．４万尾／ｍ３为宜，这种情况下，运输９．０－１１．０小时，成活率为９２．６％；运输１８．０小时，成活率８６．７％。     

菲律宾蛤仔低温保活方法的研究

本文在菲律宾蛤仔生物学研究基础上，对菲律宾蛤仔在２７．０￣－１．７℃范围内不同温度条件下的存活率，失重率及保活过程中的化学变化等进行了系统的分析研究，确定了菲律宾蛤仔的保活工艺和方法。结果表明，（１）菲律宾蛤仔的最肥季节为每年的３￣７月，出肉率（干物计）最高可达１０％；（２）最低保活温度为－１．７℃；（３）－１．０￣－１．７℃可保活１３天，存活率９１％；（４）－１．０￣１．７℃保活，保活时间最长，     

温度对中国对虾能量分配和元素组成的影响

于１９９５年８月～１０月，在２０℃、２５℃和３０℃下，以日本刺沙蚕为饵料，测定中国对虾的能量分配和元素组成（ＣＮＨ）。结果表明，水温３０℃时呼吸能显著增大（Ｐ＜０．０５），占摄食能的７７．８％。三个温度下平均能量分配比例为：排泄能２．２７％，粪便能３．４６％，蜕壳能４．１９％，生长能２２．２７％，呼吸能６７．７３％。呼吸能的变化对生长能影响显著。元素组成以２０℃含量最高，虾体能值也最大。Ｃ／Ｎ值变化表明，随温度升高，脂肪作为中国对虾能量代谢的能源物质比例增大。     

菲律宾蛤仔（Ruditapes philippinarum）低温保活方法的研究↑（*）

本文在菲律宾蛤仔（Ｒｕｄｉｔａｐｅｓ ｐｈｉｌｉｐｐｉｎａｒｕｍ）生物学研究基础上，对菲律宾蛤仔在 ２７． ０～－１． ７℃范围内不同温度条件下的存活率，失重率及保活过程中的化学变化等进行了系统的分析研究，确定了菲律宾蛤仔的保活工艺和方法。结果表明，（１）菲律宾蛤仔的最肥季节为每年的 ３～ ７月，出肉率（干物计）最高可达 １０％；（２）最低保活温度为－１．７℃；（３）－１．０～－１．７℃可保活１３天，存活率９１％；（４）－１．０～１．７℃保活，保活时间最长，失重率也低，主要化学成分无显著变化。     

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).