条纹斑竹浙江省鲨线粒体DNA的研究

用６种限制性内切酶分析了４条条纹斑竹鲨的线粒体ＤＮＡ，ＰｓｔＩ，ＨｐａＩ，ＸｂａＩ，ＥｃｏＲＩ，ＥｃｏＲＶ，ＢｇｌⅡ在条纹斑竹鲨ｍｔＤＮＡ分子上分别具有０至２个切点，ｍｔＤＮＡ分子大小为１６．６ｋｂ，根据单酶和双酶完全酶解片段的大小，构建了条纹斑竹浙江省ｍｔＤＮＡ的限制性酶切图谱。     

鲇鱼线粒体DNA的酶切图谱

采用差速离心法及ＤＮａｓｅⅠ、ＲＮａｓｅ消化法制备并纯化了鲇（Ｓｉｌｕｒｕｓａｓｏｔｕｓ）肝脏线粒体ＤＮＡ（ｍｉｔｏｃｈｏｎｄｒｉａｌＤＮＡ，ｍｔＤＮＡ）。用８种限制性内切酶对ｍｔＤＮＡ进行了分析。ＢｇⅡ、ＥｃｏｒⅠ、ＰｓｔⅠ、ＢｇｌⅠ、ＢａｍＨⅠ、ＸｂａⅠ、ＨｉｎｄⅢ、ＸｈｏⅠ在鲇ｍｔＤＮＡ分子上分别具１、１、１、２、２、７、７和０个切点。ｍｔＤＮＡ分子量约１０．８４×１０￣６道尔顿，大小为１７．５４ｋｉｌｏｂａｓｅｐａｉｒｓ。根据单酶和双酶解片段的数目和分子量，建立了鲇ｍｔＤＮＡ的限制性酶切图谱。     

湖北淤泥湖团头舫mtDNA限制性片段长度多态性的研究

用ＢａｍＨＩ、ＢｇｌＩ、ＣｌａＩ、ＥｃｏＲＩ、ＨｉｎｄⅢ、ＫｐｎＩ、ＰｓｔＩ，ＰｖｕⅡＳａｃⅡ、ＳａｃＩ、ＳｃａＩ、ＸｂａＩ，ＸｂａＩ和ＸｈｏＩ十四种限制性内切酶对来源于湖北淤泥湖的团头舫进行线粒体ＤＮＡ限制性片估长度多态性研究。初步表明ＤＮＡ上切点呈现不同程度的多态性。共检测到六种母集集团。发现一尾团头舫ｍｔＤＮＡ分子大小比普通型鱼ｍｔＤＮＡ小约０．７０ｋｂ。呈现出ｍｔＤＮＡ发子大小多态现象。     

鲢鳙线粒体DNA的九种限制性内切酶酶场图谱的比较

以十一种限制性内切酶对鲢鱼ｍｔＤＮＡ进行单酶完全酶解的切点数：ＳａｌＩ和ＢｇｌⅡ为０；ＰｓｔＩ．ＸｈｏＩ．ＫｐｎＩ和ＳａｃＩ为１；ＨｐａＩ和ＸｂａＩ为２；ＢａｍＨＩ为３；ＢｇｌＩ和ＥｃｏＲＩ为４。以九种限制性内切酶对鳙鱼ｍｔＤＮＡ进行单酶完全酶解，其切点数；ＰｓｔＩ．ＸｈｏＩ．ＳａｌＩ．ＫｐｎＩ．ＢｇｌⅡ均为１；ＨＰａＩ和ＸｂａＩ为２；ＳａｃＩ为３；ＢｇｌＩ为４。采用琼脂糖凝胶电泳法测得鲢鱼ｍｔＤＮＡ分子量为１５９３０±２２０碱基对（ｂｐ）；鳙鱼ｍｔＤＮＡ分子量为１６６５０±１５０碱基对（ｂｐ）。用单、双酶解法和部份酶解法构建了鲢鱼九种酶１８个切点和鳙鱼九种酶１６个切点的限制性内切酶酶切图谱。另外，对这两种鱼的酶解位点数，片段大小和限制酶图谱进行了比较。     

草鱼线粒体DNA限制性内切酶分析

本文报道了用ＢａｍＨⅠ，ＢｇｌⅠ，ＢｇｌⅡ，ＣｌａＩ，ＥｃｏＲＩ，ＨｉｎｄⅢ，ＰｖｕⅡ，ＳａｃⅠ，ＸｂａⅠ，ＸｈｏⅠ等十种限制性内切酶酶切草鱼线粒体ＤＮＡ，计算出各酶切片段长度及草鱼ｍｔＤＮＡ大小。其中六种酶有多个切点，在所检测的样品中未发现碱基替换。     

鲢鳙线粒体DNA的九种限制性内切酶酶切图谱的比较

以十一种限制性内切酶对鲢鱼ｍｔＤＮＡ进行单酶完全酶解的切点数：ＳａＩ和ＢｇｌⅡ为０；Ｐｓｔ Ｉ．Ｋｐｎ Ｉ和Ｓａｃ ｉ为１；Ｈｐａ和Ｘｂａ Ｉ为２；ＢａｍＨ Ｉ为３；Ｂｇｌ Ｉ和ＥｃｏＲ Ｉ为４。以九种限制性内切酶对鳙鱼ｍｔＤＮＡ进行单酶完全酶解，其切点数：Ｐｓｔ Ｉ．Ｘｈｏ Ｉ．Ｓａｌ Ｉ．Ｋｐｎ Ｉ．Ｂｇｌ Ⅱ均为１；Ｈｐａ Ｉ和Ｘｂａ Ｉ为２；Ｓａｃ Ｉ为３；Ｂｇｌ Ｉ为４。采用琼脂糖凝胶电     

华东地区中华鳖地方群体mtDNA多态分析

应用多聚酶链式反应（ＰＣＲ）技术和限制性长度多态（ＲＥＬＰ）方法，对我国华东地区中华鳖三个地方群体的线粒体ＤＮＡ（ｍｔＤＮＡ）的细胞色素（ｃｙｔ）ｂ基因进行了分析。６种限制性内切酶共获得１８种酶谱和１４种基因型。ｃｙｔｂ基因大小的估算值为１０８０±６０ｂｐ。基因型多样性和核苷酸序列多样性指数分别为０．６８７９±０．０９７８和０．０２６６±０．０９７８。浙江绍兴、江苏南京和山东青岛中华鳖群体间存在显著的遗传差异（Ｐ＜０．０５），其中南京群体与绍兴群体的遗传距离最近，青岛群体与绍兴群体的遗传距离最远。三群体的遗传关系与地理距离相一致，暗示地理分隔可能是造成中华鳖群体遗传分化的主要原因。还发现内切酶ＨｅⅢ和ＨｈａⅠ可作为中华鳖种群鉴定的标志酶。     

斑节对虾白斑综合症病毒部分基因组文库及核酸探针检测法

通过分离纯化白斑综合症病毒（ＷＳＳＶ）粒子，抽提病毒ＤＮＡ。用限制性内切酶ＥｃｏＲⅠ或ＳａｌⅠ酶切后，克隆入质粒ｐＢｌｕｅｓｃｒｉｐｔⅡＫＳ中，从而建立了ＷＳＳＶ部分基因组文库。估计ＷＳＳＶ基因组ＤＮＡ在１６５ｋｂ以上。将ＷＳＳＶ ＥｃｏＲⅠ克隆片段标记制备为探针。进行Ｓｏｕｔｈｅｒｎ杂交、打点杂交和原位杂交，其结果证明了克隆片段对ＷＳＳＶ特异，并为检测ＷＳＳＶ提供了方法。通过对部分基因组文库序列     

动物线粒体DNA研究及在鱼类种群遗传结构研究中的应用

ｍｔＤＮＡ是核外遗传物质，呈母系遗传［Ｂｒｅｓｃｈ１９８４］。由于其结构简单，易于分离，进化较快等特点［Ｂｒｏｗｎ１９７４］，在动植物种群遗传结构分析、物种及品系鉴定方面得到了广泛的应用。线粒体ＤＮＡ酶切技术在国外已被遗传学家用于鱼类种群遗传结构及品...     

建鲤线粒体DNA的遗传多态性初步分析

用16种限制性内切酶对建鲤mtDNA进行酶解分析，结果表明：建鲤的mtDNA分子大小约为16.6kb；除Bgl Ⅱ无切点，ClaⅠ、PstⅠ、SalⅠ和XhoⅠ有一个切点外，其余11种酶都有2个或2个以上酶切位点，其中DraⅠ、PvuⅡ检出mtDNA多态，将建鲤mtDNA分为两种不同的单倍型；计算建鲤种内（群体内）mtDNA多态值π＝0.00164。建鲤在分子水平显示出的遗传多样性，可能是建鲤能够适应全国各地的环境条件和养殖方式，并得以大规模推广的重要原因之一。     

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.     

Rare serotype occurrence and PFGE genotypic diversity of Streptococcus agalactiae isolated from tilapia in China

Previously, we reported 10 PEGE types of 85 tilapia Streptococcus agalactiae(GBS), which shifted from Streptococcus iniae in China, by using PEGE method. Presently, larger and more representative tilapia GBS were isolated, for the ?rst time in China, to characterize their serotypes and genetic diversities more precisely than had done before. 168 GBS strains were distributed in ?ve provinces of China, in which Guangdong, Guangxi and Hainan were the major ones, holding36.9%(62/168), 37.5%(63/168) and 19.6%(33/168), respectively. Serotypes, Ia, Ib and III, were observed in these strains and the most predominant one was Ia(95.2%), which mainly distributed in Guangdong, Guangxi and Hainan. Ia initially occurred in 2009, it shoot up to 32.1% in 2010,but decreased to 16.1% in 2011 before went up to 45.2% in 2012. Ib sporadically occurred during2007–2011, III onlyoccurred in 2012. 14 different PFGE types, including 4 new types(N, O,P and Q), were observed, in which B, D, F and G were the predominant types, holding 83.9%(141/168) of the total GBS strains. Ia corresponded to 11 PFGE types(A–H, N–P), in which type D predominated(51%). Ib represented 3 genotypes(I, J and Q) and III harbored only 2genotypes(N and F). Type N and Fsynchronously presented in Ia and III. In summary, the genetic diversity of tilapia GBS varied by serotypes and changed with geographical locations and years.Although Iastillpredominated, new rareserotypeIII alreadyoccurred in China.     

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.     

Purification and partial characterization of GtHs (cfLH and cfFSH) from Indian walking catfish (Clarias batrachus) (L.) and development of a homologous ELISA for cfLH

Two gonadotropins (GtH; Qa and Qb) were purified by gel filtration and ion exchange chromatography from the pituitaries of Indian walking catfish (Clarias batrachus). The presence of GtH during purification was assessed by in vitro oocyte maturation and in vivo steroidogenic activity, and their identities were determined by elution profiles, molecular weight, biological activities and yield. The molecular weights of Qa and Qb were 37 and 42 kDa, respectively, and composed of distinct subunits (Qa: 20 and 14 kDa and Qb: 26 and 18 kDa). Polyclonal antibodies raised against Qa immunostained Qa, Qb and pituitary GtH cells. A competitive Qa‐ELISA was developed whose sensitivity was 6.25 ng mL^?1 (1.25 ng well^?1) with intra‐ (3.5%) and inter‐ (12.4%) assay coefficients of variation. Displacement curves parallel to the standard were obtained with plasma and pituitary extracts of catfish, Qb and carp GtHII. The assay was validated by measuring the plasma Qa levels after LHRH treatment and in relation to ovarian growth in the female catfish during different reproductive phases. Based on the results, Qa and Qb corresponded to fish LH and FSH respectively. The findings will increase the knowledge of the mechanisms controlling fish reproduction and identification of sensitive phases in fish in captivity for hormonal manipulation.     

Controlled infection of Poecilia reticulata Peters (guppy) with Tetrahymena by immersion and intraperitoneal injection

Tetrahymena is a protozoan parasite, which infects guppy, Poecilia reticulata Peters, and causes substantial economical losses in commercial farms worldwide. Studies of guppy infected by Tetrahymena require standardized infection protocols. The LD50 for Tetrahymena infection of guppies by intraperitoneal (IP) injection was calibrated, and the level obtained was 946 parasites per fish. Guppy infection with Tetrahymena by immersion, imitating the natural route of infection via the integument, was studied under normal or stress conditions. Exposure to cold and netting (CNI) and to cold only (CI) followed by immersion exposure to 10 000 Tetrahymena per mL resulted in 22.5% and 19.2% mortality, respectively, as compared to 14.2% and 10% in groups that were netted only (NI) or non‐stressed (I). Histopathology revealed that immersion infection resulted in a systemic infection. Lysozyme levels, measured 3 weeks after infection, were significantly higher in the CNI group (288 μg per mg protein) compared with CI‐, NI‐ and I‐treated groups (94.5, 64 and 62.3 μg mg^?1, respectively). There was no evident parasite immobilization activity in body homogenates, suggesting no development of acquired immunity. Re‐infection by IP injection revealed no increase in protection in any of the treatment groups, mortality range of 56.3–75%, higher than in the non‐exposed control (40.6% mortality).     

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.