Effects of biofloc technology on reproduction and ovarian recrudescence in Nile tilapia

The objectives of this study were to evaluate the reproductive cycle, morphological changes of ovary and mobilization of energy reserves in Nile tilapia reared with biofloc technology (BFT). In general, the growth and reproductive performance were highly similar between BFT and Control system (clear water). Difference between the systems was found in the hepatosomatic index (using mixed‐effects models), which suggested that BFT can alter the energy mobilization in the post‐spawning period. The absolute and relative fecundity, fertilization rate, number of larvae produced per female, gonadosomatic index, proportion of oogenesis cells, number of post‐ovulatory and atretic follicles were similar between the two systems. We also did not detect a reduction in the reproductive cycle length in Nile tilapia reared in BFT. Because there was no evidence of the negative effects of BFT on Nile tilapia reproduction, we concluded that BFT might be used for breeder stocking of this species.     

泰山螭霖鱼(Varicorhinus macrolepis)在年周期中卵巢发育的组织学研究

本文对泰山螭霖鱼(Varicorhinus macrolepis)卵巢的组织结构在年周期中的变化规律进行了研究。结果表明: 泰山螭霖鱼卵母细胞的发育过程在年周期中分为6个时相。卵巢发育分期依卵巢切面中所占面积超过50%或居最高比例的卵母细胞的时相来划分,分期情况如下:2-4月份,卵巢由Ⅳ期发育至第Ⅴ期,4月份进入繁殖期;4-6 月份,为第Ⅴ期;6月下旬-7月份为第Ⅵ期,是排卵后退化阶段;8月份,为Ⅱ期卵巢(休整期);9月份-翌年的1 月份为第Ⅲ期,并以Ⅲ期卵巢越冬。由研究结果可知:泰山螭霖鱼为短期分批产卵类型,一年产卵一次。     

长鳍吻鮈性腺发育的初步观察

采用常规石蜡切片技术对野生长鳍吻鮈(Rhinogobio ventralis(Sauvage et Dabry))性腺进行了组织学观察,描述了其各类型生殖细胞形态及卵巢、精巢的分期特征。结果显示:长鳍吻鮈卵巢为细线状或扁条状,精巢为细线状或直棍状;卵细胞发生经历了卵原细胞、初级卵母细胞和次级卵母细胞3个阶段,精细胞发生经历了精原细胞、初级精母细胞、次级精母细胞、精子细胞及精子5个阶段;性腺发育分为6个时期,卵巢中第Ⅱ时相晚期至Ⅲ时相早期的初级卵母细胞中出现核仁外排现象;同一时期卵巢中存在多种时相的卵母细胞,故推测其卵巢发育类型为部分同步型,精巢为小叶型结构。     

用3种方法提取中华绒螯蟹卵巢总蛋白效果的比较

为了探寻适于中华绒螯蟹Eriocheir sinensis卵巢蛋白质组学研究的总蛋白最佳提取方法,采用RC DC Protein Assay检测蛋白浓度、SDS-PAGE分析蛋白组成等,比较分析了用裂解液法Ⅰ、裂解液法Ⅱ和TCA/丙酮沉淀法提取中华绒螯蟹卵巢蛋白的效果。结果表明:应用3种方法提取的卵巢总蛋白浓度分别为15.59、13.05、0.65 mg/m L,用裂解液法提取的效果最佳,而TCA/丙酮沉淀法不适合用于中华绒螯蟹卵巢总蛋白的提取;SDS-PAGE电泳图谱进一步显示,用裂解液法Ⅱ提取的蛋白样品组成较为丰富,且重复性好,比较适合后续蛋白质组学研究。该研究结果可为中华绒螯蟹卵巢发育蛋白质组学研究提供参考。     

牛MEN1基因真核表达载体的构建及其表达分析

多发性内分泌腺瘤致病因子1 (multiple endocrine neoplasia Ⅰ,MEN1)参与乳腺发育与泌乳行为的调控.本研究克隆了牛(Bos taurus) MEN1基因(bMEN1)的全长cDNA,并在不同细胞中检测bMEN1mRNA及其编码蛋白menin的表达情况.根据GenBank中bMEN1基因序列,设计特异性引物,用qRT-PCR方法得到附带EcoR Ⅰ和HindⅢ酶切位点的bMEN1片段,并将其克隆到真核表达载体pcDNA3.1-myc-his(A)中.体外转染物种同源性细胞牛乳腺上皮细胞(bovine mammary gland epithelial cell,MAC-T)和异源性细胞中国仓鼠(Cricetulus barabensis)卵巢细胞(Chinese hamster ovary cells,CHO)、小鼠(Mus musculus)成肌细胞(mouse myoblast cells,C2C12),利用qRT-PCR和Westem blot技术检测转染前后bMEN1 mRNA和蛋白menin的表达.双酶切和基因测序结果表明,成功构建了bMEN1基因的真核表达载体pcDNA3.1-myc-his-bMEN1.所建立的转染体系可以使目的基因bMEN1在3种不同细胞中成功表达mRNA和目的蛋白,转染后24 h都达到最高表达量,并达到极显著水平(P＜0.01),之后逐渐降低.其中,CHO细胞中的转染24 h后bMEN1 mRNA和menin蛋白的表达量分别是对照的28 415倍和5.65倍.本研究建立了bMEN1基因的真核表达载体及其转染体系,能够在不同细胞中成功表达,为体外研究MEN1基因对于乳腺的调节功能及其对于机体代谢的调节机制提供了一种工具和技术体系.     

绵羊RARG基因在发情不同阶段卵巢中的mRNA表达

【目的】分离鉴定绵羊视黄酸受体-γ（Retinoic acid receptor gamma,RARG）基因,分析该基因的分子特征,研究其在发情周期不同阶段卵巢中的表达差异。【方法】选取年龄和体况相近、健康的甘肃高山细毛羊周岁母羊8只,通过同期发情处理,根据卵巢生理状态,将其分为两组：黄体期组和卵泡期组,各4只,屠宰后,采集卵巢组织,以周岁母羊卵巢组织RNA为模板,采用RT-PCR技术扩增绵羊RARG基因并克隆到pMD18-T载体中进行测序验证。利用ORF Finder和DNAStar等生物信息学软件分析绵羊RARG基因的理化性质、同源性和预测结构域和功能。利用Q-PCR技术,检测绵羊RARG基因在母羊发情周期不同阶段卵巢组织中的相对表达量,用SPSS19.0软件进行差异显著性分析,探讨RARG基因对绵羊发情周期的调控机制。【结果】获得了绵羊RARG基因包含完整编码区序列在内的1 588 bp的绵羊RARG基因序列,并提交至NCBI,GenBank登录号为KF019682。该序列含有1个1 377 bp的完整开放阅读框（Open reading frame,ORF）,编码458个氨基酸,其蛋白分子质量为50 955.8 D,理论等电点为7.45。绵羊RARG基因与牛、普通狨、家犬、仓鼠、家猫、人、小鼠、虎鲸、狒、家鼠、野猪、海牛、海豚和蟾蜍等14个物种的同源性分析,结果显示RARG基因在上述物种间高度保守,且与牛的亲缘关系较近,序列同源性最高。GO功能分析结果显示,绵羊RARG基因作为结构蛋白参与转录调控的几率最高,分别为0.272和0.223。在绵羊RARG基因编码的氨基酸序列上发现了2个功能结构域,分别是核激素受体c4锌指结构域（c4 zinc finger in nuclear hormone receptors,ZnF_C4）和 激素受体配体结构域（ligand binding domain of hormone receptors,HOLI）。Q-PCR结果显示,绵羊RARG mRNA 在甘肃高山细毛羊卵泡期卵巢中的相对表达量显著高于黄体期（P<0.05）。【结论】绵羊RARG基因在物种间高度保守,含有ZnF_C4和HOLI等2个与转录调控相关的功能结构域,推测该基因可能作为一种重要的转录因子参与母羊发情周期调控。     

Surgical resection of a dysgerminoma in a mare

A mare was referred for further evaluation of a mass found in the left caudal abdomen during a routine postpartum reproductive palpation. The mare was clinically normal with no history of health problems. Ultrasonographic examination of the mass confirmed its presence, but the origin of the mass could not be accurately determined. Routine haematology and serum biochemistry results were within normal limits. The mare was initially treated conservatively with antibiotics, but the mass continued to increase in size, so it was surgically excised. The mass involved the left ovary. The mare showed transient abdominal pain after surgery, but developed no other complications and was in foal 7 months later. On histology, the mass was diagnosed as a dysgerminoma, a rare ovarian tumour of germ cell origin.     

Undifferentiated tumor in the ovary of a corn snake (Elaphe guttata guttata)

A 6-year-old intact female corn snake (Elaphe guttata guttata) was presented with a 3-week history of anorexia. Coelomic radiographs revealed a 9 x 4 cm soft tissue opacity suggestive of a right ovarian enlargement. The mass (9 x 5 x 4 cm) was surgically removed, and multiple smears from tissue sections were stained with Diff-Quik. Multiple tissue samples from the mass were collected and fixed in formalin. Cytologic specimens were moderately cellular and contained light pink amorphous background material. The cells were primarily spindle-shaped with moderate to marked anisocytosis and anisokaryosis. Cells sometimes were round to polygonal, and rarely were arranged in small clusters. Macrophages occasionally were observed. Histologic specimens consisted of a highly cellular mass composed of pleomorphic, spindle-shaped cells and, occasionally, round to polygonal cells arranged in irregular fascicles. The neoplastic cells were immunoreactive for cytokeratin (AE1/AE3), smooth muscle actin, and skeletal muscle actin, but did not stain for vimentin or desmin. On the basis of the morphologic and immunohistochemical results, a diagnosis of ovarian undifferentiated carcinoma was made. In this report, we describe the challenges of using immunohistochemistry to diagnose this uncommon type of tumor in reptiles.     

野生斑节对虾卵巢发育过程中不同组织的氨基酸组成

测定了海南岛南部海域野生斑节对虾（Penaeus mofbodon）亲虾在卵巢发育不同阶段的全虾、肝胰腺和卵巢的氨基酸组成。结果表明,在卵巢发育初级阶段（含I期和Ⅵ期）、中级阶段（含Ⅱ期、Ⅲ期和Ⅳ期）和高级阶段（V期）斑节对虾全虾样品中,牛磺酸（Tau）质量分数差异显著（P〈0．05）,由高到低为中级阶段〉高级阶段〉初级阶段;而其他的氨基酸质量分数、氨基酸总量和必需氨基酸总量略有变化,但差异不显著（P〉0．05）。肝胰腺和卵巢氨基酸总量随着性腺发育逐渐上升,临产时（V期）下降,但差异不显著（P〉0．05）。中级阶段卵巢中的天门冬氨酸（Asp）、谷氨酸（Glu）、脯氨酸（Pro）、酪氨酸（Tyr）和赖氨酸（Lys）等5种氨基酸质量分数处于峰值。