基于线粒体COⅠ基因序列的中国沿海缘边银鲈遗传多样性分析#br#

为了解中国沿海缘边银鲈(Gerres limbatus)的遗传背景,分析了9个地理群体140条样本线粒体细胞色素氧化酶Ⅰ(cytochrome oxidase subunitⅠ,COⅠ)基因5′端652 bp序列。结果发现14个变异位点,13个单倍型,其中有9个独有单倍型和4个共享单倍型,整体单倍型和核苷酸多样性都较低(H_d=0.276±0.050,π=0.0005±0.0001)。琼州海峡以东海域的遗传多样性(H_d=0.338~0.456,π=0.0006~0.0011)高于琼州海峡以西北部湾海区(H_d=0~0.295,π=0~0.0005)。单倍型网络图未出现明显谱系结构和地理结构。群体遗传分化系数F_st显示,硇洲群体与其他群体间出现不显著的中低度遗传分化(F_st=0.0773~0.1696,P>0.05),其他群体间分化不明显(F_st=-0.0415~0.0612,P>0.05),总体上群体间分化程度都不高。AMOVA分析显示,遗传变异绝大部分(95%以上)来源于群体内。中性检验结果显示,群体总体的Tajima’s D(-2.2549)和Fu’s F_s(-16.4725)结果均为显著性负值(P<0.01),核苷酸错配图呈现较为明显单峰,单倍型网络图呈典型星状结构,表明缘边银鲈历史上存在过种群扩张,扩张时期约为距今0.1304~0.0435百万年。由于总体单倍型多样性小于0.5,核苷酸多样性小于0.005,推测中国缘边银鲈种群曾经历过较为严重的瓶颈效应。宁德、硇洲群体的核苷酸多样性相对较高,应优先保护。北部湾各群体遗传多样性极低,需要采取措施避免遗传多样性下降。     

中国近海军曹鱼线粒体细胞色素b基因序列的遗传变异

测定了连云港、舟山、防城群体34 ind军曹鱼(Rachycentron canadum)线粒体细胞色素b基因883 bp序列,共检测到5个变异位点,发现6个单倍型,平均单倍型多样性(h)和核苷酸多样性(π)分别为0.324和0.000 4,总体表现出较低的遗传多样性;其中连云港群体遗传多样性最高,单倍型多样性和核苷酸遗传多样性分别为0.473±0.162和0.000 57±0.005 93;而舟山群体没有任何变异。连云港与舟山、防城群体间的FST值分别为0.029(P=0.00)与0.042(P=0.00),舟山与防城群体间的FST为-0.048 03(P=0.00),表明连云港与其它两个群体间仅有较低的分化而舟山与防城群体间无明显分化。分子方差分析(AMOVA)表明,3个群体的遗传变异大部分来自于群体内(74.45%,P=0.000)。军曹鱼单倍型拓扑结构呈星状排列,将3个群体作为一个整体进行Tajimas D和Fu’s Fs分析,二者均为显著负值(FST=-1.922 40,P﹤0.00;FST=-5.735,P﹤0.00),表明军曹鱼在历史上经历了种群的扩张,根据τ的观察值0.364,估算出军曹鱼种群扩张时间约为3.1~1.2万年,即末次冰盛期。     

基于线粒体COⅠ的南海北部长棘银鲈遗传多样性分析

为了解中国长棘银鲈(Gerres filamentosus)的遗传背景以更好地保护与开发利用种质资源,测定了南海北部7个群体线粒体COⅠ基因5′端序列,以分析其遗传多样性和遗传结构。结果发现,176尾长棘银鲈652 bp序列中有25个单倍型,155个变异位点。7个群体整体呈现较高的单倍型多样性(H_d=0.767±0.018)与较低的核苷酸多样性(π=0.055 0±0.003 7)。就遗传多样性指数而言,碣石群体(H_d=0.606±0.092,π=0.059 2±0.006 6)最高,次高为阳江群体(H_d=0.323±0.102,π=0.027 0±0.010 7);琼海群体(H_d=0.560±0.125,π=0.001 0±0.000 3)最低,次低为陵水群体(H_d=0.685±0.077,π=0.001 5±0.000 3)。群体遗传分化系数F_(st)显示,碣石、阳江群体与其他群体间存在高度(极显著)的遗传分化(F_(st)=0.517 1～0.851 4,P<0.001),这2个群体间有低等程度(显著)的遗传分化(F_(st)=0.111 1<0.15,P<0.05),而其他群体间无明显分化(F_(st)=-0.025 5～0.008 8,P>0.05)。AMOVA分析显示,群体变异主要来源于群体内个体间(61.69%～74.13%),但群组间仍有较多变异(19.06%～37.92%);原因可能与历史上琼州海峡的阻碍、当代复杂的海流以及长棘银鲈只进行短距离洄游等生态特性有关。中性检验显示,海南陵水、东方和新盈群体在晚更新世时期发生过种群扩张,但南海长棘银鲈总体未呈现种群扩张现象,可能是不同谱系的叠加造成整体核苷酸不配对分析图呈现多峰。碣石和阳江群体与其他群体间有高度的遗传分化,可将它们作为一个独立的管理保护单位,其中碣石群体遗传多样性最高,应重点保护;其他群体可作为另一管理保护单位,其中琼海和陵水群体遗传多样性较低,建议及时采取保护措施,避免遗传多样性过度下降而导致资源枯竭。     

基于线粒体COI序列的中国沿海蓝点马鲛遗传多样性

为了解中国沿海蓝点马鲛(Scomberomorus niphonius)的遗传背景以更好地保护和开发利用种质资源,对分布于渤海、黄海、东海和南海海域的6个群体76 ind蓝点马鲛线粒体COI基因712 bp序列进行了测定,并结合Gen Bank中下载的9条南海蓝点马鲛序列,分析其遗传多样性、种群结构和历史动态。共检测到21个变异位点,17个单倍型,总体呈现高单倍型多样性(Hd=0.702±0.044)和低核苷酸多样性(π=0.002 8±0.000 2)的特点,其中渤海、黄海和东海海域的蓝点马鲛群体遗传多样性指数相对较高(Hd:0.695~0.816,π:0.002 7~0.003 3),而南海群体明显偏低(Hd=0.442±0.145,π=0.001 7±0.000 6),推测是由于黄海和东海是中心分布区,而南海是边缘分布区的缘故。AMOVA分析结果显示,群体间(-6.27%~-1.52%)不存在变异,群体内个体间(101.52%~106.56%)的变异是变异的主要来源;群体间遗传分化系数Fst值为-0.138~0.040(P0.05),遗传距离N_m值为-81.145~-4.134、11.876~146.559,均大于4或小于0,表明群体间不存在遗传分化,不同海域群体间基因交流频繁,这可能与蓝点马鲛分布范围广、具长距离迁移能力、产卵和越冬时均可发生洄游以及鱼卵具漂浮性且为多次性产卵类型等原因有关。聚类分析的邻接树与单倍型网络图上出现的2个分支均在更新世晚期发生过种群快速扩张事件,但蓝点马鲛总体在数据上未呈现出种群扩张现象,可能是2个分支的叠加造成整体核苷酸不配对分析图呈现多峰分布。     

基于线粒体控制区序列的南海北部近岸鯻的遗传多样性

80尾鯻(Therapon theraps)样本采自广东碣石、阳江,广西东兴,海南临高、陵水,测定其线粒体控制区951 bp序列,发现42个变异位点,60个单倍型。南海北部近岸鯻总体的单倍型多样性(0.986±0.007)和核苷酸多样性相对较低(0.009±0.000),其中陵水群体核苷酸多样性最高(0.010),碣石、阳江、东兴群体最低(0.008)。单倍型网络图与邻接树没有形成明显不同的谱系分支和地理聚群,表明不存在谱系结构和地理结构。大陆沿岸碣石、阳江和东兴群体间的遗传分化系数(Fst)为–0.017 5~0.008 9(P0.05),基因流(Nm)值为–14.54~27.84;海南岛临高和陵水群体间Fst为–0.046 0(P0.05),Nm为–5.68,表明大陆沿岸群体内部和海南岛群体内部都无明显分化;但海南岛与大陆群体间的Fst为0.061 6~0.135 3(P0.05),Nm为1.87~3.81。分子方差分析显示,大陆沿岸群体和海南岛群体间分化极显著(FCT=0.100 7,P=0.000 0),可能是受大陆与海南岛分离形成的琼州海峡,全年余流方向总趋势是由东向西,且强度较高的影响。南海北部近岸鯻可作为2个管理单位进行保护,建议优先保护陵水群体。单倍型简约性网络图呈非典型的星状分布,中性检验Fu’s Fs为显著负值且核苷酸不配对分析为单峰分布,表明历史上经历过种群扩张事件,推测扩张年代为晚更新世(3.62万年前),扩张后的有效群体大小约为扩张前的8.7×106倍,可能是受到海退缩减和海侵扩张的影响。     

华南6水系与澜沧江-湄公河攀鲈线粒体ND2基因的遗传多样性分析

测定了中国华南6水系及澜沧江(云南勐腊)-湄公河流域(柬埔寨洞里萨湖)的125尾攀鲈(Anabas testudineus)线粒体部分ND2基因1 010 bp序列,分析发现39个变异位点和12个单倍型,总遗传多样性较低(h=0.369,π=0.003 8),推测可能经历过严重的瓶颈效应;中国攀鲈群体遗传多样性更低(h=0.282,π=0.000 4),处于边缘区的中国攀鲈群体是造成低遗传多样性的主要原因。在单倍型网络图中柬埔寨和中国攀鲈各自聚类,具有明显地理结构和谱系结构,推测地质运动和气候变化导致基因交流受阻所致。核苷酸错配图和中性检验表明中国群体经历过种群扩张,时间约为(5.94~4.13)万年前。华南水系群体间基因交流通畅,不存在明显分化;但与云南澜沧江群体间分化大而显著(FST=0.775,P0.01),AMOVA分析显示变异主要来自组群间(77.41%),推测二者分化时间约为(4.0~2.8)万年前,云南群体受末次冰期的影响,基因交流受阻而出现分化。中国群体和柬埔寨群体可作为2个管理单位进行保护;就中国群体而言,韩江水系群体遗传多样性最高,建议优先保护;澜沧江与华南水系间群体分化显著且遗传多样性极低,建议对澜沧江水系群体进行保护,以避免种质资源灭绝。     

基于线粒体COⅠ基因序列的江苏省4个太湖新银鱼群体遗传多样性分析

为了解江苏省主要湖泊内太湖新银鱼(Neosalanx taihuensis)的遗传背景,利用PCR扩增江苏省太湖、高邮湖、洪泽湖和骆马湖4个群体153尾样本的COⅠ基因序列,并进行测序和分析。经比对获得630 bp的基因序列片段,检出7个变异位点,其中简约位点3个,单一位点4个。153尾个体共定义9个单倍型,单倍型多样性为0.580±0.022(0.050±0.047～0.361±0.103),核苷酸多样性为0.001 06±0.000 07(0.000 08±0.000 07～0.000 62±0.000 20),呈现出较低的单倍型多样性和核苷酸多样性的特点。4个群体间的遗传距离为0.000 12～0.001 83。AMOVA分子方差分析表明,太湖新银鱼群体间的遗传差异(76.84%)大于群体内遗传差异(23.16%),遗传变异主要来自群体间。种群间遗传分化系数统计检验表明,太湖和洪泽湖群体及高邮湖和骆马湖群体之间差异不显著。中性检验和歧点分布分析表明,4个太湖新银鱼种群偏离中性进化,历史上经历过种群扩张。研究可为太湖新银鱼种质资源科学保护和合理利用提供理论依据。     

基于细胞色素b基因河鲈养殖群体与野生群体遗传多样性分析

利用线粒体DNA细胞色素b基因的421 bp部分序列对北屯、乌伦古湖、博斯腾湖3个野生河鲈(Perca fluviatilis)群体和北湖、五家渠2个养殖河鲈群体序列多样性与种群遗传结构进行分析。结果表明:100个个体中检测到7个单倍型,变异位点10个,其中野生群体60个个体检测到9个变异位点和6个单倍型,养殖群体40个个体共检测到8个变异位点和4种单倍型;野生群体平均单倍型多样性和平均核苷酸多样性(Hd=0.496±0.121,Pi=0.002 53±0.001 54)高于养殖群体(Hd=0.416±0.127,Pi=0.001 23±0.001 13),分子方差分析揭示,98.74%的遗传变异性出现在种群内个体间。群体间的FST分析揭示野生群体和养殖群体分化程度较低(0.05FST0.15)。分子系统树和单倍型网络图分析也表明,河鲈单倍型间关系较近,野生群体和养殖群体不存在显著分化。     

香鱼野生群体和养殖群体遗传多样性比较

香鱼是分布于中国、日本和朝鲜的一种珍稀名贵经济鱼类,本实验比较分析了香鱼养殖和野生群体的遗传多样性。研究结果显示,在长度为445 bp的控制区部分序列上,鳌山卫养殖群体的单倍型多样度h(0.198 4±0.092 4)和核苷酸多样度π(0.000 8±0.000 9)显著低于东张水库野生群体(h=0.810 5±0.067;π=0.002 6±0.002 0),两群体产生了较大的遗传分化(F st=0.447,P=0);单倍型邻接关系树的拓扑结构简单,未呈现明显的地理谱系结构,日本香鱼个体与中国香鱼亲缘关系较远;东张群体的历史动态分析结果表明其可能经历过近期的群体扩张事件。无论是养殖群体还是野生群体,中国香鱼群体的遗传多样性现状不容乐观。     

基于线粒体COⅠ基因序列的中国近海蓝圆鲹遗传多样性研究

为了解蓝圆鲹(Decapterus maruadsi)遗传多样性和种群结构,提出合理的蓝圆鲹种质资源保护依据,测定并分析了中国沿海10个地理群体193个体线粒体细胞色素C氧化酶I(COⅠ)基因序列。在652 bp的COⅠ序列中共检测到62个变异位点,29个单倍型,总体呈现较高的单倍型多样性(0.626 00±0.030 00)和低核苷酸多样性(0.002 25±0.001 16)的特点。核苷酸多样性显示:以福建海域为界北方高于南方(北方π=0.004 06～0.005 68,南方π=0.000 17～0.003 05),南海以阳江为界东部高于西部(东部π=0.001 43～0.005 68,西部π=0.000 17～0.002 86),单倍型网络图未出现明显系谱结构和地理结构。群体间遗传分化系数F_(st)显示,北海群体与其他群体有较高的遗传分化(F_(st)=0.145～0.798,P<0.001),新盈与陵水、惠来、阳江、福州和舟山样本之间也存在较高的遗传分化(F_(st)=0.006～0.229,P<0.05),其余群体间分化不显著。AMOVA分析表明,组群内群体间的变异比例占到16.66%～19.49%,原因可能是蓝圆鲹只作短距离洄游的生活习性以及受中国沿岸流、黑潮、暖流和北部湾特殊的大陆架的影响、北海和新盈与其他群体基因交流有限所致。蓝圆鲹整体的Fu’s F_S为显著性负值(F_S=-1.171 8,P<0.01);核苷酸错配峰图为明显单峰;单倍型网络图呈典型星状结构,表明蓝圆鲹群体经历过种群扩张。根据Bayesian skyline plot(BSP)分析扩张大约发生在0.222百万年前。北海和新盈群体与其他群体间存在较高的分化,可作为独立的管理保护单位,其中核苷酸多样性最低的新盈群体需避免遗传多样性的进一步降低;其余群体作为另一管理保护单位,其中崂山群体核苷酸多样性最高,舟山群体其次,应重点保护二者的种质遗传资源。     

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).