Optimal conditions for egg storage, incubation and post-hatching growth for the freshwater turtle, Chelodina rugosa: Science in support of an indigenous enterprise

Incubation of northern snake-necked turtle (Chelodina rugosa) eggs and subsequent sale of hatchlings for the pet industry has the potential to provide culturally suitable employment for indigenous communities in northern Australia. Developmental arrest in response to egg inundation is unique to C. rugosa. Eggs can be stored under water for up to 10 weeks without appreciable impact on egg or embryo survival, allowing the transport and sale of eggs into niche markets without high levels of mortality, and permitting eggs to accumulate in diapause until there are sufficient numbers to incubate as batches. Eggs that are not inundated or inundated for short periods experience similar survival rates to eggs inundated for lengthy periods. Incubation temperature influences embryo survival and development period in C. rugosa. Embryonic survival is greatest at 26 °C, steadily declining as temperature increases to 32 °C. A similar increase in incubation temperature decreases incubation period by approximately 40 days, however almost half of this variation is attributed to the increase in incubation temperature from 26 to 28 °C. Hatchling growth in C. rugosa is characterized by two phases. There is an initial phase of relatively slow growth under the partial influence of initial egg size and incubation duration, followed by a second phase of relatively rapid growth under the partial influence of water temperature and mass at hatching. Post-hatching survival is negatively correlated with duration of egg inundation and water temperature. Evidence suggests that inundation of C. rugosa eggs for 6 weeks, incubation of embryos at 28 °C and raising hatchlings in 28 °C water will yield the best overall outcomes.     

鲁西黄牛耳缘组织成纤维细胞的体外培养

实验以鲁西黄牛耳缘组织为研究材料,采用组织块贴壁培养法对其进行了成纤维细胞的体外培养。通过细胞的传代培养、形态学和动力学观察与分析,结果表明,所建立的鲁西黄牛耳缘组织成纤维细胞系具有典型的成纤维细胞形态,胞体均为梭形和不规则三角形,且细胞丰满;细胞核形态为卵圆形,并位于胞质中央,核仁清晰。在细胞生长于繁殖过程中亦经历了潜伏期、指数增生、停滞期和衰退死亡期等4个时期。细胞群体倍增时期为72h。实验结果表明,已成功地培养了鲁西黄牛耳缘组织成纤维细胞。     

鲶繁殖生物学的研究

嘉陵江鲶的卵母细胞划分为６个时相。第Ⅱ时相卵母细腻外不仅具有质膜，而且还有滤泡膜的结缔组织膜，第Ⅳ时相卵终细腻具漏斗状和精孔及椭圆形的精孔细腻。嘉陵江鲶的成熟年龄大多为１龄。生殖期雌雄经为２：１。繁殖季节中雌鱼的成熟系数高达１４．５％。对鲶进行人工催产，成功地获得了鲶的受精卵。鲶的受精卵圆形、绿色，膨大后其直径为．０５－４．５７ｍｍ。温度在２７．５－３１℃，幼鱼孵出需要２９个小时３０分钟。初孵仔鱼     

鸡Mx基因全长cDNA序列的克隆及分析

以Poly(I)-Poly(C)诱导鸡成纤维细胞Mx基因的表达,提取总RNA,RT-PCR扩增出全长Mx cDNA,扩增产物克隆入pMD19-T Simple载体中进行序列测定。结果表明:与GenBank公布的鸡Mx cDNA序列相比,其同源性达99.9%,为进一步研究鸡Mx基因的抗病毒活性和作用机理奠定了基础。     

日本米虾胚胎发育及离体孵化

本研究采用自制离体孵化装置,对日本米虾(Caridina japonica)不同发育期胚胎进行离体孵化研究,结果显示,水温为25.5℃时,日本米虾受精卵孵化大约需要25 d,发育积温为637.5℃。胚胎发育历经受精卵、卵裂期、囊胚期、原肠胚期、前无节幼体期、后无幼体期、前溞状幼体期和膜内溞状幼体期8个时期。各期离体胚胎均能孵化出幼体,膜内溞状幼体期离体胚胎孵化率最高,为(80.7±2.4)%,非离体孵化的对照组为(79.1±4.9)%,二者差异不显著;卵裂期离体胚胎孵化率最低,为(28.2±2.6)%,与对照组差异显著。各组离体胚胎所孵化出的Ⅰ期(ZⅠ)和Ⅱ期溞状幼体(ZⅡ)的变态率无显著差异。温度对日本米虾前溞状幼体期胚胎离体孵化影响显著,在15.0℃~32.5℃范围内,随水温升高孵化时间逐渐缩短,15.0℃时,前溞状幼体离体孵化时间为(436.8±124.8) h,32.5℃时缩短至(228.0±88.8) h,但温度高于29.0℃时,孵化出的幼体变态率开始下降。本研究可为日本米虾繁殖生物学及甲壳动物胚胎离体孵化技术研究提供参考。     

供体细胞对猪体细胞克隆胚胎早期发育的影响

以中国农业大学实验用小型猪香猪胎儿成纤维细胞、成年耳成纤维细胞和颗粒细胞3种细胞系为供体细胞进行核移植。比较了血清饥饿法和接触抑制法处理胎儿成纤维细胞诱导进入G0／G1期的效率，发现二者差异不显著（P〉0．05），血清饥饿2d和4d差异不明显，同样接触抑制2d和4d差异也不显著（P〉0．05）。系统研究了影响克隆胚胎发育的供体因素：血清饥饿与否、细胞形态、细胞类型及个体差异等，结果表明：血清饥饿处理对克隆胚的早期发育没有明显的促进作用；圆形光滑细胞有利于细胞融合，对早期发育无显著影响（P〉0．05）；不同个体、不同类型的供体细胞对克隆胚囊胚发育率有一定的影响。     

鲁西黄牛体细胞克隆保种技术的研究

以体外培养的鲁西黄牛耳皮肤成纤维细胞作核供体，研究利用体细胞克隆技术保存我国地方黄牛优良品种的技术方法。通过组织块培养法建立的鲁西黄牛（1♂，4♀）成纤维细胞系，作为核移植供体细胞，利用屠宰场母牛卵巢卵母细胞经体外培养成熟后作为核受体进行核移植，试验结果表明：重构胚的融合率为62．5％（242／387），分裂率为63．6％（154／242），体外培养第7天囊胚发育率为42．9％（66／154）。体外培养第7天的囊胚的内细胞团细胞（ICM）与滋养层细胞数平均为37和47，ICM占44．2％。体外发育到7d的囊胚新鲜胚胎的移植妊娠率（新鲜胚胎）移植受体10头，60d妊娠率为20％（2／10）。     

分子修饰前后人参皂苷体外抗马立克氏病毒鸡胚成纤维细胞模型的建立

用雏鸡将马立克氏病毒(MDV)血毒复壮,分离发病鸡淋巴细胞并接种于鸡胚成纤维细胞,观察其病变。获得适应鸡胚成纤维细胞(CEF)的MD强毒,通过电镜观察、琼脂扩散实验进一步鉴定病毒,并建立了MDV感染CEF细胞模型,为研究人参皂苷及其衍生物的体外抗病毒作用及其机制奠定基础。     

Analysis of chromosome aberration due to ethidium bromide using AFM

AIM: To study chromosome aberration due to ethidium bromide (EB),a heterocyclic organic compound and an organic fluorescence dye commonly used in biochemical experiment,and to help further understanding the molecular mechanism of tumor or cancer induced by EB and other heterocyclic organic compounds.METHODS: The toxicity action of EB was evaluated from three aspects including DNA,chromosome and embryo stem cells (ESCs) using atomic force microscopy (AFM),and thereinto,the morphology structural difference of ESCs treated with two EB doses was also valuated.RESULTS: The morphological structures of DNA,chromosome and ESCs were dramatically damaged.The average height of DNA decreased 0.5 nm;chromosomal arms were ruptured from centromere location;molecules of cellular membrane congregated and loop-like structure formed,and ES cell masses were collapsed and became dead after large EB doses treatment and mesh-like morphological structure was discernable.CONCLUSION: The toxicity action of EB is strong and destroys the surface structure of DNA and chromosome.EB induces structural aberration of ES cellular membrane and cell death.The results indicate that the action of EB is externalized at gene level and cell level,which is important to study the carcinogenicity of EB.     

家蚕胚胎伴性温敏性的遗传研究

试验用新九与伴1等4个温敏性的品种杂交，将其F1代蚕种用高温干燥条件催青，结果雄蚕正常孵化，雌蚕几乎不孵化，表明伴1等品种具有控制催青温敏性表达的基因存在。用华1与伴1组配的F1、F2及F1与亲本回交的14个组合作遗传分析，认为催青敏感性状由位于Z性染色体上的一个主基因控制，呈隐性的伴性遗传。