NaCl盐度和NaHCO3碱度对鲤、鲫和大鳞鲃的精子活力及其受精率的影响

研究了不同NaCl盐度(0、1、2、3、4、5、6、7、8、9、10、11、12)和NaHCO₃碱度(0 mmol/L、10.00 mmol/L、15.85 mmol/L、25.12 mmol/L、39.81 mmol/L、63.10 mmol/L)对松浦镜鲤(Cynipus carpio)、方正鲫(Carassius auratus gibelio)和大鳞鲃(Barbus capito)精子活力及其受精率的影响。结果表明: 1) 在盐度为4时, 方正鲫和大鳞鲃精子的激烈运动时间、快速运动时间和寿命最长, 方正鲫分别为(90.11±9.03) s、(126.34±13.90) s和(154.27±11.36) s; 大鳞鲃分别为(48.91±1.43) s、(62.19±4.28) s和(90.68±4.46) s。在盐度为5时松浦镜鲤精子的激烈运动时间、快速运动时间和寿命最长, 分别为(72.44±9.42) s、(102.16±8.82) s和(206.99±6.65) s。2) 当盐度达到8以上时, 3种鱼的精子激活将受到抑制; 盐度大于10时, 方正鲫和大鳞鲃精子死亡; 盐度大于11时, 松浦镜鲤精子死亡。3) 在碱度为15.83 mmol/L时, 3种鱼的精子激烈运动时间、快速运动时间和寿命均最长, 且显著高于其他碱度条件下的精子活力(P<0.05)。4) 在盐度为1时, 方正鲫和大鳞鲃受精率达到最高分别为63.0%和68.0%, 在盐度为3时, 松浦镜鲤受精率达到最高为72.3%, 当盐度大于3时, 受精率开始呈明显下降趋势。碱度为10.00 mmol/L时, 3种鱼的受精率均最高, 分别为75.4%、54.0%和66.0%。本实验旨在通过测定不同NaCl盐度、NaHCO₃碱度条件下3种鱼类精子的活力和受精率所受的影响, 为北方地区碳酸型盐碱水域的渔业开发利用提供基础资料。 

     

建鲤和黑龙江野鲤自交和正反交F1形态差异分析

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P₁为76.7%~96.7%, 判别准确率P₂为85.2%~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。

     

Molecular cloning and expression of kidney‐type glutaminase from common carp (Cyprinus carpio) and its up‐regulation by glutamine in primary culture enterocyte

Glutaminase (GLS) is the key enzyme of glutamine (Gln) metabolism and utilization. In this study, a cDNA encoding GLS protein was identified from common carp Cyprinus carpio intestine. The open reading frame of GLS cDNA encodes a polypeptide of 595 amino acids, which shows a high similarity with its zebrafish Danio rerio counterpart. Bioinformatic analysis showed the protein belongs to kidney‐type GLS. The putative protein has glutaminase domain and ankyrin repeats domain, which are highly conserved among vertebrate orthologues. Real‐time quantitative PCR analysis revealed that the abundance of GLS mRNA was the highest in the white muscle, followed by the brain, eyeball and pituitary. Glutaminase was ubiquitously expressed in all intestinal segments of common carp. The activity of GLS did not distribute uniformly along the entire length of the intestine. In primary culture enterocyte, and the expression of GLS mRNA is up‐regulated quickly and effectively by Gln.     

氯氰菊酯通过食物链在鲤鱼肾脏和小肠中的生物积累效应

以鲤鱼为研究对象,对常用农药氯氰菊酯通过食物链（蔬菜-黄粉虫幼虫-鲤鱼）在鲤鱼肾脏和小肠内的富集效应进行了研究。饲喂的蔬菜农药残留剂量为2．00mg·kg^-1和10．00mg·kg^-1,通过HPLC定量检测黄粉虫幼虫体内及鲤鱼肾脏和小肠内氯氰菊酯的富集浓度,同时对鲤鱼肾脏和小肠组织结构进行了观察。结果显示,氯氰菊酯在黄粉虫幼虫及鲤鱼体内的富集浓度与剂量呈正相关,在食物链中的富集呈逐级递增,且对黄粉虫幼虫及鲤鱼的摄食情况及活动能力有明显的影响,随着处理时间的延长及剂量的增大,氯氰菊酯对鲤鱼肾脏及小肠结构损伤程度也增大。因此,杀虫剂氯氰菊酯的使用会通过食物链对环境中生物乃至人类健康和生态平衡造成影响,应慎重使用。     

Cloning, nucleotide sequence and phylogenetic analyses, and tissue-specific expression of the transferrin gene in Cirrhinus mrigala infected with Aeromonas hydrophila

Transferrin partial complementary DNAs were cloned from the livers of five species in four genera of Indian carps (Indian major carp species: Labeo rohita, Catla catla and Cirrhinus mrigala; medium carp: Puntius sarana; minor carp: Labeo bata) subsequent to polymerase chain reaction amplification with published heterologous primers or self-designed primers derived from conserved regions of transferrin cDNA sequences. The partial transferrin cDNAs of the five species of carps had sizes from 624 to 633 bp (487 bp for L. rohita) and encoded an open reading frame consisting of 206–211 (162 for L. rohita) amino acids. The alignments of carp cDNA sequences showed 85–97% homology and 71–93% homology in deduced amino acid sequences. A phylogenetic tree of amino acid sequences of transferrin cDNAs from carps showed that the relationship among the four genera of Indian carps is well correlated with that derived from classic morphologic analyses. The hypothesized cleavage site and interdomain bridge of transferrin molecule were predicted for the above carp species and interestingly the cleavage site amino acid sequence was found to be conserved among all the carps. To study the tissue-specific expression of the transferrin gene, various tissues (liver, kidney, spleen, brain, muscle, testis, heart, intestine, gill and fin) from apparently healthy (control), moribund and survived C. mrigala experimentally infected with Aeromonas hydrophila infection were analyzed. Transferrin mRNA was detected only in liver RNA and to lesser extent in brain tissue out of the 10 tissues analyzed irrespective of bacterial infection.     

构建鲤现代种业的核心技术

对于任何一个农业物种而言,现代种业核心技术包括3个方而：收集与创制最优良的种质资源、建立先进实用的育种技术、方便的可最大化利用上述资源的数据库及网站。针对鲤这个水产养殖种,黑龙江水产研究所及合作单位经过30多年的研究积累和创制了抗寒能力强、生长速度快、肉质好的三大系列种质,在此基础七已经培育出6个水产新品种;构建了基于亲本遗传距离的分子育种技术,并在常规育种技术的使用过程中增加了统计分析和染色体操作技术,如BLUP技术、雌核发育技术等,将现代育种技术充分地运用于鲤的品种培育之中;同时建瓿了涵盖表型、生物学特征、一般遗传参数的鲤种质数据库和涵盖基因、标记及经济性状QTLs的鲤基因组数据库,使鲤具备了现代种业需要的生物学、遗传学、基因组学的完整数据库。综上所述,初步构建了鲤现代种业的核心技术。     

Seasonal Differences in Growth Performance of Four Strains of Common Carp,Cyprinus carpio in India

To assess strain related differences in growth performance and growth patterns under the same culture environment, four strains of common carp, two each of the scale carp, Cyprinus carpio var. communis (Chinese big-belly carp and long bodied carp) and mirror carp, C. carpio var. specularis (scattered carp and linear carp) were communally stocked in three fertilized earthen ponds of 0.14 ha each at 5,000 fish ha^?1 in the ratio of 1:1:1:1 during an 11-month (February to December) culture cycle. Chinese big-belly carp grew larger than other groups, among which there were no significant differences. Scale carp strains performed relatively better than mirror carp at higher temperatures and then essentially stopped growing as temperatures declined into winter. The strains of mirror carp, on the other hand continued growing well later into the cold season.     

Effects of myo‐inositol on proliferation,differentiation, oxidative status and antioxidant capacity of carp enterocytes in primary culture

This study investigated the effects of myo‐inositol (MI) on the growth and antioxidant capacity of carp enterocytes. The enterocytes were incubated in media containing 0, 15, 30, 45, 60 and 75 mg MI L^?1 for 96 h. The results indicated that MI could increase cell viability. In addition, the activities of cellular alkaline phosphatase (AKP), gamma‐glutamyl transpeptidase (γ‐GT), Na⁺, K⁺‐adenosine trisphosphatase (Na⁺, K⁺‐ATPase) and creatinkinase (CK) increased with MI supplementation at levels ranging from 15 to 60 mg MI L^?1 medium, indicating an improvement in cell differentiation and function. Further, enzymatic antioxidant ability, as measured by total superoxide dismutase (T‐SOD), Cu/Zn‐SOD, Mn‐SOD, catalase (CAT), glutathione peroxidase (GPx) and glutathione‐S‐transferase (GST) activities, improved with MI supplementation. Finally, cell damage, as indicated by lactic acid dehydrogenase (LDH) activity, malondialdehyde (MDA) content of the medium and cellular protein carbonyls (PC), was all depressed by MI. Correlation analyses showed that cell viability (MTT) was positively related to the antioxidant enzyme activities, but negatively related to cell damage (LDH, MDA and PC). In summary, the data showed that MI could improve the growth of fish enterocytes. This result may be partly due to the enhanced antioxidant status and depressed oxidative damage.     

建鲤生长激素促泌素受体1基因的特性及其与增重相关SNP位点的筛选

GHS-R基因在哺乳类为候选的肥胖和生长数量性状位点。本实验使用RT-PCR及PCR的方法,从建鲤(Cyprinus carpio var.jian)分离到2个jlGHS-R1s,称为jlGHS-R1a和jlGHS-R1b(分别简称1a和1b)。jlGHS-R1s的阅读框编码360个氨基酸,两者间相似性高达96%;另外还存在2个jlGHS-R1s的转录变体(分别简称1a’和1b’),选择性切割了翻译起始碱基后第490-680 nt的191 bp片段,该片段两端碱基分别为gt-ag,这种选择性切割导致翻译提前终止,仅翻译184个氨基酸,包括前面3个半跨膜区,这与已报道的罗非鱼等的保留内含子的转录变体不同。jlGHS-R1s的阅读框被1个内含子分隔,该内含子位于第260个氨基酸密码子的第1和第2个碱基之间,1a和1b的内含子长度分别为676 bp和885 bp。使用分段PCR在建鲤群体的1a和1b基因上共找到32个SNP位点,构建PCR-RFLP法对其中9个SNPs进行基因型检测,结果发现各种基因型在322尾建鲤中均有出现,但存在着明显的偏分布。与增重的相关分析表明,位点1a-I_C386T和1b-E1_G159T与雌雄鱼鱼种和成鱼阶段增重分别呈极显著(P<0.01)和显著(P<0.05)相关,CC型和GG型个体增重最快,另外有5个位点则与其中的某阶段或者某性别的增重相关,为了确定上述所得标记的适用性,选取其中4个标记在另外7个家系共610尾鱼中检测,结果 C386T和G159T还是与增重呈极显著(P<0.01)和显著(P<0.05)相关,可用于建鲤分子育种。     

Lipid peroxidation,protein oxidant and antioxidant status of muscle and serum for juvenile Jian carp (Cyprinus carpio var. Jian) fed grade levels of methionine hydroxy analogue

Lipid peroxidation, protein oxidation and antioxidant status of serum and muscle in juvenile Jian carp (Cyprinus carpio var. Jian) fed graded levels of methionine hydroxy analogue (MHA: 0, 5.1, 7.6, 10.2, 12.7, 15.3 g kg^?1 diet) for 60 days were investigated. Both malondialdehyde and protein carbonyl content in serum and muscle decreased with increasing dietary MHA level up to 5.1–10.2 g kg^?1 diet (P < 0.05). Anti‐hydroxyl radical and activities of catalase, glutathione peroxidase and glutathione reductase in muscle and serum, as well as anti‐superoxide anion, superoxide dismutase activity and glutathione content in serum, increased with optimal MHA supplement (P < 0.05). Meanwhile, glutathione‐S‐transferase activity in serum showed a downward trend with dietary MHA up to 7.6 g kg^?1 diet (P < 0.05). These results indicated that MHA improved antioxidant status and depressed lipid and protein oxidation in serum and muscle.