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1.

建立牙鲆(Paralichthys olivaceus)雌核发育近交系和杂交系, 以两性生殖家系作为对照组, 3个群体的遗传特征和生长进行比较。结果表明, 雌核发育近交系、杂交系和对照组的等位基因数分别为415578, 平均等位基因数为1.72.33.3, 平均观测杂合度为0.419 20.654 90.916 7, 雌核发育杂交系的上述参数明显高于近交系, 但低于对照组。3个群体的平均纯合度分别为0.580 70.345 10.083 3, 个体之间的遗传相似度分别为0.831 20.826 10.672 7, 杂交系小于近交系, 但明显高于对照组(P<0.05)。在生长方面, 雌核发育杂交系比近交系和对照组生长快, 差异极显著(P<0.01)。从结果可以看出, 雌核发育杂交系具有比近交系高的杂合度, 但具有与近交系相似的遗传相似度; 生长上比近交系明显快, 表现出明显的杂交优势。结论认为雌核发育家系杂交方法可以作为一种育种手段, 用于发挥杂交优势和提高遗传相似度。

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2.

在实验室条件下, 设计相同的牙鲆(Paralichthys olivaceus)鱼苗网箱养殖密度(密度5/, 鱼苗体质量约21 g), 分别与不同数量的双齿围沙蚕(Perinereis albuhitensis, 0 g, 50 g, 70 g, 90 g)混合养殖, 4个实验组分别记作CS1S2S3, 网箱规格为Ф60 cm×30 cm。各实验组设3个平行组, 实验进行40 d。牙鲆和沙蚕在称重前均饥饿24 h, 同时取5尾牙鲆和20 g沙蚕作为实验初始样品。实验期间, 每天07:3018:002次投喂配合饲料, 根据对照组饵料剩余情况适时调整投喂量, 每个实验单元的投饵量基本一致, 此过程中不处理残饵和粪便。实验单元采用微流水, 流量为250~500 L/d。实验期间, 24 h持续微量充气。至实验结束时, 全部生物饥饿24 h。将其全部取出, 擦干体表水分, 称重后置于70℃干燥箱中烘干至衡重, 研磨、分析。结果表明, 对照组(C)的底质积累碳、氮量最高, 各实验组水体净输出的氮和有机碳量以及呼吸碳量分别无显著差异(P>0.05)。饲料是碳、氮输入的主要来源。在输出各项中, 各个实验组水体净输出氮占输出氮比例无显著差异(P>0.05), 而系统净排放氮以对照组所占比例最高, 但与S1S2组无显著差异(P>0.05); 对照组底质积累氮所占输出氮比例显著高于S2S3(P<0.05)。对照组底质积累碳所占比例显著高于其他各实验组(P<0.05); 对照组和S1组的水体净输出有机碳比例显著高于S3(P<0.05), S3组碳净排放占碳输出的比例显著低于对照组和S1(P<0.05)。碳、氮产投比均以对照组最低。在本研究条件下, 双齿围沙蚕在综合养殖模型中可利用牙鲆网箱养殖产生的残饵和粪便, 减少底质碳、氮积累, 提高产投比, 综合养殖模型既可以降低碳、氮排放, 又可以增加产品产出。结论认为, 网箱养殖牙鲆搭配双齿围沙蚕的综合养殖模型理论上具有可行性, 双齿围沙蚕可以作为牙鲆网箱养殖产生的残饵和粪便的一道有效处理屏障, 此模型在港湾和围堰鱼类网箱养殖中更有意义。

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3.

在草鱼(Ctenopharyngodon idella)养殖系统中维持碳氮比为201、水温(26.0±2.3)pH7.2~7.8, 24 h不间断供氧以形成生物絮团, 监测培养过程中养殖水体总氮(TN)、总固体悬浮物(TSS)浓度、碱度的动态变化及分析生物絮团的营养组分, 并应用PCR-DGGE技术研究生物絮团的原核及真核微生物组成和动态变化。养殖水体TN变化范围为6.65~11.15 mg/L, 9天达到峰值(10.11±1.05) mg/L, 12天后和第0天时的TN水平无显著性差异(P>0.05); TSS度在第9天达到最大值(419.67±11.5) mg/L, 15天后TSS稳定维持在244.67 mg/L; 碱度变化范围为136.68~239.20 mg CaCO3/L, 6天达到峰值后逐渐下降并趋于平稳。生物絮团的粗蛋白含量为30%(干重)生物絮团原核微生物主要由变形菌门(Proteobacterium)放线菌门(Actinobacteria)、拟杆菌门(Bacteroidetes)和某些未知不可培养的细菌组成, 蓝细菌(Cyanobacterium)只存在于生物絮团培养前期(0510), 产碱菌科细菌(Alcaligenaceae)存在于生物絮团形成的整个过程, 且是第51015天的优势菌。组成生物絮团的真核微生物隶属于原生动物门的斜叶虫属(Loxophyllum)、隐藻纲的隐鞭藻科(Cryptomonadaceae)Goniomonas、硅藻纲的双头菱形藻属(Nitzschia)。其中双头菱形藻属为絮团培养初始第0天到15天特有; 斜叶虫属在絮团培养后期分布较多。结论认为, 生物絮团系统在养殖的15 d左右达到稳定运行的状态, 能有效调节养殖系统菌藻分布, 控制养殖水质, 维持整个系统的平衡与良性发展。

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4.
青虾血蓝蛋白基因的克隆、表达分析及多克隆抗体的制备   总被引:3,自引:2,他引:1  

应用RACE技术克隆了青虾(Macrobrachium nipponense)Hc基因全长cDNA序列, 并对该基因序列特征进行了分析。青虾Hc基因cDNA全长2 235 bp, 包括10 bp5′末端非翻译区(UTR), 2 074 bp的开放阅读框(ORF), 151 bp3′UTR, 开放阅读框编码688个氨基酸。蛋白相似度比对显示, 青虾血蓝蛋白含有典型的6个保守的铜离子结合位点。系统进化树分析表明, 青虾Hc与脊尾白虾(Exopalaemon carinicauda) Hc聚在一起, 具有最近的亲缘关系。荧光定量PCR检测显示, Hc基因在青虾不同组织中均有表达, 其表达量在肝胰腺中最高; 使用荧光定量PCR检测青虾Hc基因在低氧胁迫和复氧条件下在肝胰腺中的mRNA时空表达情况, 结果显示, 经低氧和复氧刺激后, 与对照组相比, Hc 在肝胰腺中的表达量分别在低氧胁迫 12 h24 h 和复氧6 h出现了3次明显上调, 由此推测Hc基因参与低氧应激分子过程。此外, 本研究通过血蓝蛋白的分离纯化技术制备了多克隆抗体, 本研究结果可为更进一步的了解青虾血蓝蛋白的生理功能提供理论支撑。

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5.
大菱鲆选育家系抗鳗弧菌性能   总被引:5,自引:2,他引:3  

基于家系选育技术开展了大菱鲆(Scophthalmus maximus)抗鳗弧菌病选育研究, 2010年构建的37个选育二代家系中选择成活率高的30个家系进行鳗弧菌(Vibrio anguillarum)感染实验, 开展对鳗弧菌的抗病性研究。结果表明, 不同家系间抗鳗弧菌感染能力存在着显著差异(P0.05); 30个家系中, 鳗弧菌感染后, 12个家系的存活率达到65% 以上, 其余家系的存活率则低于65%; 通过bCOX回归分析, 计算各家系的优势比, 优势比最高的5个家系的存活率达到65% 以上; 各家系的成活率、优势比和死亡历时差4项指标进行聚类分析, 优势比最高且存活率达到65% 以上的5个家系聚为一类。综合感染家系的高成活率、高优势比以及聚类分析的结果, 选育出5个抗病力较强的优良家系。选育出的抗病力较强的家系可做为抗鳗弧菌选育的核心育种群体, 为抗鳗弧菌病的传代选育奠定良好的基础。

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6.

(25±0.5) ℃, 设置4个水流速度实验组, 分别为静水对照组0 bl/s(体长/)1.0 bl/s组、2.0 bl/s组和4.0 bl/s, 研究不同水流速度对中华倒刺鲃(Spinibarbus sinensis)幼鱼肌肉自由基代谢的影响, 训练周期为8周。结果显示, 肌肉超氧化物歧化酶(SOD)和谷胱甘肽过氧化物酶(GSH-PX)的活性随着水流速度的增加而升高, 训练组过氧化氢酶(CAT)活性出现不同程度的下降, 而抗氧化物谷胱甘肽(GSH)的含量未发生明显变化(P>0.05); 训练组肌肉总抗氧化能力(T-AOC)均高于对照组, 2.0 bl/s组最高, 4.0 bl/s组次之, 1.0 bl/s组最低, 2.0 bl/s4.0 bl/sT-AOC与对照组有显著性差异(P<0.05)8周训练后, 2.0 bl/s组丙二醛(MDA)含量显著低于其他3(P<0.05), H2O2含量随着水流速度增加呈上升趋势8周运动训练对骨骼肌B细胞淋巴瘤因子-2 (Bcl-2)Bcl-2相关X蛋白(Bax)的含量及血清肌酸激酶(CK)活性无显著影响(P>0.05), 2.0 bl/sBaxBax/ Bcl-2显著低于4.0 bl/s(P<0.05)。结果表明, 2.0 bl/s的水流速度可提高中华倒刺鲃肌肉的抗氧化能力, 降低脂质过氧化水平, 并在一定程度上抑制肌细胞凋亡。

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7.

通过测定建鲤(Cyprinus carpio var. jian)和黑龙江野鲤(Cyprinus carpio haermatopterus)自交和正反交组合的形态参数, 运用方差分析和多元分析, 比较了4个自交和正反交组合的形态差异。结果表明, 不同组合间在侧线鳞、侧线上鳞、侧线下鳞、胸鳍鳍棘以及腹鳍鳍棘数量上没有显著差异(P>0.05)。而在背鳍鳍棘、背鳍鳍条、腹鳍鳍条、臀鳍鳍棘、臀鳍鳍条以及胸鳍鳍条数量上有差异(P<0.05)。聚类分析结果表明, 自交组合之间的形态最为接近, 两个杂交组合与两个自交组合的趋异程度逐渐增大。用判别分析方法构建了4个判别方程, 判别准确率P176.7%~96.7%, 判别准确率P285.2%~90.6%, 综合判别率为87.5%。利用主成分分析提取了9个主成分, 对总变异的累积贡献率为68.943%。研究结果为建鲤和黑龙江野鲤及其杂交后代的种群和亲缘关系鉴定以及育种提供了科学依据。

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8.
斑鳜精液超低温冷冻保存及其效果分析   总被引:1,自引:1,他引:0  

实验所用翘嘴鳜(♀, Siniperca chuatsi) 2~3, 体质量1 000~1 500 g; 斑鳜(♂, Siniperca scherzeri Steindachner)1~2, 体质量300~500 g。于繁殖季节选取成熟度好的亲鱼以促黄体素释放激素类似物(LHRH-A2)、绒毛膜促性腺激素(HCG)和马来酸地欧酮(DOM)进行催产。通过筛选D-15D-17Ringer M-Hank’s4种稀释液和二甲基亚砜 (DMSO) 甘油(Gly)、乙二醇 (EG)1,2-丙二醇 (PG)和二甲基甲酰胺 (DMF)5种抗冻剂, 发现DMSOD-17分别是优选的抗冻剂和稀释液。以D-17作为斑鳜精子的稀释液, 按照13(精液稀释液)体积比稀释, 添加10%体积的DMSO作为抗冻剂, 按照三步冷冻法超低温冷冻保存, 37水浴解冻的斑鳜精子, CASA分析精子活率为(83.26±18.20)%, SCGE检测表明70%以上的精子核DNA不会发生损伤; FCM分析表明26.74%的解冻精子有完整的细胞膜且线粒体功能正常; 用冷冻复苏斑鳜精子与翘嘴鳜精卵进行人工授精, 最高受精率(39.6±6.5)%, 温度24~28孵化时间38 h, 孵化后的鱼苗发育正常, 开口1周以后的鱼苗全长(1.346±0.255 ) cm, 体高(0.438±0.103) cm, 体质量(0.045±0.020) g。鲜精受精鱼苗和冻精受精鱼苗在开口后1周的生长没有显著差异。因此认为D-17+10% DMSO可用于斑鳜精液的超低温冷冻保存。本研究将有助于斑鳜种质资源的收集保存和冷冻保存的斑鳜精液在翘嘴鳜(♀)×斑鳜(♂)杂交中的应用。

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9.

选择在中国分离得到的一株能产生麻痹性贝毒(Paralytic Shellfish Poison, PSP)的赤潮甲藻塔玛亚历山大藻(Alexandrium tamarense) ATHK, 研究其对中国明对虾(Fenneropenaeus chinensis)鳃组织氧化胁迫和Caspase基因(FcCasp)表达的影响。将中国明对虾分别暴露于200 cells/mL 1 000 cells/mL塔玛亚历山大藻中, 于胁迫后361224487296 h测定鳃的超氧化物歧化酶(SOD)活性、谷胱甘肽硫转移酶(GST)活性、丙二醛(MDA)含量和FcCasp基因相对表达量, 以不加藻的过滤海水作为对照。结果表明, 200 cells/mL塔玛亚历山大藻胁迫下, SOD活性、GST活性、MDA含量和FcCasp相对表达量均随取样时间推移表现出先上升后下降的趋势。而在1 000 cells/mL塔玛亚历山大藻胁迫下, SOD活性随时间增加呈现先升高后降低的趋势, 24~96 h 被显著抑制(P<0.05), GST活性除348 h外均被显著抑制(P<0.05), MDA含量和FcCasp相对表达量均整体表现上升的趋势, 呈现出一定的时间效应。相关性分析显示, 塔玛亚历山大藻胁迫下, 中国明对虾鳃的MDA含量和FcCasp相对表达量呈正相关(P<0.05)。实验结果表明, 塔玛亚历山大藻可破坏中国明对虾氧化抗氧化系统的平衡, 引发对虾鳃组织的脂质过氧化(LPO), 造成其氧化损伤, 从而导致FcCasp表达的上调。本实验结果为将SOD活性、GST活性和MDA含量作为生物标志物, 用于评价塔玛亚历山大藻胁迫提供了依据, 同时也为研究塔玛亚历山大藻对中国明对虾的危害机制提供了参考。

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10.
基于微卫星标记对长江中上游胭脂鱼增殖放流效果的评估   总被引:2,自引:1,他引:1  

利用已经发表的11对胭脂鱼多态性微卫星标记, 对长江中上游胭脂鱼(Myxocyprinus asiaticus)增殖放流效果进行评估。结果表明, 5个胭脂鱼养殖场内所有的149尾繁殖亲本和长江中上游采集的65尾胭脂鱼中共观测到140个等位基因, 其观测杂合度和期望杂合度的平均值(范围)分别为0.771(0.519~0.906)0.759(0.469~0.894), 多态信息含量(范围)0.726(0.392~0.882)。通过软件Cervus统计得到, 11个微卫星座位累计排除率为99.998 3%, 并且在长江中上游采集的65个样本中, 11个样本与养殖场内繁殖亲本确定存在亲子关系, 据此确定这11尾为增殖放流的胭脂鱼, 并由此推算增殖放流的胭脂鱼对长江中上游野生群体的贡献量为16.92%。研究结果表明, 增殖放流是实现胭脂鱼野生种群资源恢复的有效途径。

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11.
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.  相似文献   

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14.
Growth hormone (GH) and reproduction: a review   总被引:5,自引:0,他引:5  
Interaction between growth and reproduction occurs in many vertebrates and is particularly obvious at certain stages of the life cycle in fish. Endocrine interactions between the gonadotropic axis and the somatotropic axis are described, the potential role of GH being emphasised. A comparative analysis of these phenomena in mammals, amphibians and fish, suggests a specific role of GH in the physiology of puberty, gametogenesis and fertility. It also shows the original contribution made by studies on the fish model in this field of investigations.  相似文献   

15.
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.  相似文献   

16.
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).  相似文献   

17.
The purpose of this work was to evaluate the suitability of a method based on the presence of two restriction sites (for Hae III and Hindf I) in the mitochondrial NADH dehydrogenase subunit 5 (mt ND5) gene to identify Scomber species. The evaluation was performed on 144 reference and market samples by sequencing of the entire 505-bp fragment of the mt ND5 gene and of a 464-bp fragment of the Kocher fragment of the cytochrome b gene (mt Cytb). Sequence analysis of any of the two fragments allows the identification of each of the four Scomber species, but S. japonicus and S. colias had the same restriction sites at the ND5 amplicon and would not have been differentiated by this analysis. Similarly, loss of the Hae III site in some S. scombrus individuals would have misidentified them as not being Scomber. All the market products were correctly labeled except one acquired in Spain labeled as originating in the Atlantic and containing S. japonicus.  相似文献   

18.
There has been growing concern about the overuse of antibiotics in the ornamental fish industry and its possible effect on the increasing drug resistance in both commensal and pathogenic organisms in these fish. The aim of this study was to carry out an assessment of the diversity of bacteria, including pathogens, in ornamental fish species imported into North America and to assess their antibiotic resistance. Kidney samples were collected from 32 freshwater ornamental fish of various species, which arrived to an importing facility in Portland, Oregon from Colombia, Singapore and Florida. Sixty‐four unique bacterial colonies were isolated and identified by PCR using bacterial 16S primers and DNA sequencing. Multiple isolates were identified as bacteria with potential to cause disease in both fish and humans. The antibiotic resistance profile of each isolate was performed for nine different antibiotics. Among them, cefotaxime (16% resistance among isolates) was the antibiotic associated with more activity, while the least active was tetracycline (77% resistant). Knowing information about the diversity of bacteria in imported ornamental fish, as well as the resistance profiles for the bacteria will be useful in more effectively treating clinical infected fish, and also potential zoonoses in the future.  相似文献   

19.
This study investigated the efficiency of iodophor disinfection (135 ppm active iodine for 15–30 min) of non‐hardened Salmo trutta eggs against different groups of bacteria and against fungus. Egg samples were taken from non‐disinfected and from disinfected eggs, microorganisms were cultured on specific nutrient media and their mass was measured by turbidimetric methods. Bacteria and fungus mass of non‐hardened eggs could be reduced but not eliminated by iodophor disinfection with 135 ppm active iodine for 15 min. The extent of reduction was 47–65% (Experiment 1). The efficiency of disinfection increased with disinfection time as the reduction in bacteria and fungus mass was 40–55% after 15 min and 58–74% after 30 min (Experiment 2). Disinfection efficiency of iodophor solution diluted in water (reduction 49–57%) and of iodophor solution diluted in sodium chloride solution iso‐osmolar to the oocytes (reduction 52–61%) was similar (Experiment 3). The reduction in bacteria and fungus mass was persistent as it was 39–72% lower in embryos deriving from disinfected eggs than in embryos deriving from non‐disinfected ones (Experiment 4). In conclusion, the tested disinfection method is inadequate to eliminate pathogens completely but it could positively influence immune defence of eggs and embryos.  相似文献   

20.
The endemic, anadromous cyprinidChalcalburnus tarichi is the only fish species known to occur in alkaline Lake Van (Eastern Anatolia, Turkey). EightC. tarichi were maintained individually in Lake Van water (17 – 19°C; pH 9.8; 153 mEq·I–1 total alkalinity; 22 total salinity) and tank water samples analyzed for 24 h in 2 to 4 h intervals. At zero time, < 1µM ammonia was present and urea was undetectable in the tank water; at 24 h, total ammonia and urea made up 114±32 and 35±25µM, respectively. Over the experimental period, ammonia-N and urea-N excretion averaged 1041±494 and 607±169moles·kg–1 fish·h–1, respectively. The extent of urea excretion was highly variable between specimens. Uric acid excretion was not detectable.Urea was present at high concentrations in all tissues and plasma (25 – 35moles·g–1·ml–1) of freshly caughtC. tarichi; total ammonia content of the tissues was by a factor of 1.9 (liver) to 3.0 (brain) lower. High arginase activity (2.4±0.2 U·min–1·g–1) was detected in the liver ofC. tarichi but ornithine carbamoylphosphate transferase, a key enzyme of the ornithine-urea-cycle, was absent. Ureagenesis is likely through degradation of arginine and/or uricolysis. High glutamine synthetase activity (11±0.6 U·min–1·g–1) and low ammonia content in brain suggest that, like other teleosts,C. tarichi has an efficient ammonia detoxification in the brain, but in no other tissue.Nitrogenous waste excretion at alkaline pH is discussed. The ability ofC. tarichi to excrete high levels of ammonia at extremely alkaline pH is unique among teleosts studied so far. The mechanism of ammonia excretion under Lake Van conditions remains to be elucidated.  相似文献   

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