-
Identification and expression analysis of goose type lysozyme in half smooth tongue sole (Cynoglossus semilaevis)
作 者: Zhen-Xia Sha, Qi-Long Wang, Yang Liu, Song-Lin Chen 摘 要: Lysozymes are considered to be potent innate immune molecules against the invasion of bacterial pathogens. The goose-type lysozyme is one of the three major distinct lysozyme types identified in the animal kingdom including teleosts. In this report, we identified, sequenced, and characterized the goosetype lysozyme gene (CsGLys) from half-smooth tongue sole (Cynoglossus semilaevis). The full-length cDNA of CsGLys is 1191 bp in length from the transcription start site to polyadenylation site, including a 91 bp 50-terminal untranslated region (UTR), a 452 bp 30-terminal UTR and a 648 bp open reading frame (ORF) of encoding a polypeptide with 215 amino acids. The deduced amino acid sequence of CsGLys possesses a Goose Egg White Lysozyme (GEWL) domain with three conserved residues (E91, D104 and D121) essential for catalytic activity. The CsGLys gene consisting of 2535 bp, was similar to those of other teleost species such as Japanese flounder and large yellow croaker with five exons... -
MHC polymorphism and disease-resistance to Edwardsiella tarda in six turbot (Scophthalmus maximus) families
作 者: DU Min, CHEN SongLin, LIU YanHong, NIU BaoZhen, YANG JingFeng & 摘 要: This study examined genetic variation in the major histocompatibility complex (MHC) Class II B gene in turbot (Scophthalmus maximus) by virulent bacterial pathogen challenge. One hundred fry from each of six families were infected with Edwardsiella tarda by intraperitoneal injection. Family mortality ranged from 28.0% to 83.3%. Complete exon 2 and intron 1 sequences of MHC Class II B genes were amplified from five survivor and five non-survivor individuals per family using the clone-sequence method. Thirty-seven sequences from 60 individuals revealed 37 different alleles, 25 of which were unique to this study. The 25 unique alleles belonged to 16 major allele types. Nine alleles were used to examine the association between alleles and resistance/susceptibility to disease. Five alleles were present in an individual, suggesting a minimum of three loci or copies of the turbot MHC Class II B gene. The rate of non-synonymous substitution (dN) was 2.30 and 1.58 times higher... -
牙鲆胚胎冷冻损伤的观察
作 者: 王春花 ,田永胜 摘 要: 为了观察牙鲆胚胎在冷冻保存过程中的形态受损情况,将其浸入20%PM(20%丙二醇和20% 甲醇1:l的混合液)中平衡,并用程序化法和玻璃化法对其冷冻保存2h后解冻,用摄影显微镜记录其在抗冻剂里平衡时和冷冻保存后的形态。结果显示在平衡过程中胚胎卵膜出现凹陷(称为溶液损伤),但可以恢复;在冷冻过程出现胞内冰损伤,是致命的;用程序化法冷冻保存的尾芽期胚胎卵膜和卵黄完好,胚体边缘受伤;用玻璃化法保存的尾芽期胚胎,卵膜和卵黄损伤较重,胚体损伤较轻;因此将玻璃化法和程序化法结合可以达到扬长避短的效果。 -
牙鲆胚胎程序化冷冻保存研究
作 者: 王春花,陈松林 ,田永胜 ,刘本伟 ,邓寒 摘 要: 以牙鲆胚胎为材料 ,对胚胎程序化冷冻保存的主要环节进行了探讨 ,以优化牙鲆胚胎的程序化超低温冷冻保存法。单一抗冻剂的毒性试验表明甲醇(Methanol) 、 1 ,22丙二醇( PG)的毒性低于其他抗冻剂的毒性。 -
牙鲆胚胎玻璃化冷冻技术研究
作 者: 田永胜,陈松林,严安生 摘 要: 对名优经济鱼类牙鲆 (Paralichthys Oliuaceus) 胚胎的玻璃化冷冻技术方法进行了研究。筛选出了适合于牙鲆胚胎平衡和培养的基础液 BS2, 对牙鲆不同时期胚胎在玻璃化液 VS2中的耐受能力进行了研究 -
牙鲆胚胎玻璃化冷冻及解冻过程中的温度因子与玻璃化率
作 者: 田永胜,刘本伟,陈松林 摘 要: 为避免鱼类胚胎玻璃化冷冻过程中脱玻璃化现象发生,本文对冷冻过程中温度变化及解冻方法进行了研究。在室温13℃下,将含有玻璃化液FVS3的麦管直接投入到一196℃ 的液氮中,麦管内温度达到一196℃ 时,所用的时间为15.46±1.38 s,降温速率为800。C/rain。对玻璃化麦管在35~45。C水浴中解冻时间进行测定。结果显示,解冻所需时间在6.17~7.94 S之间,麦管从一196℃升高到13。C的解冻速率为1 500~2 000。C/rain。当水浴温度从37℃逐步升高到72。C时,对水浴表面蒸气温度的变化进行了测定,相应水温下蒸气温度从25.47℃升高到38.6。C。在麦管中加入不同浓度的玻璃化液FVS1、FVS2、FVs3和FVS4进行玻璃化冷冻,然后将麦管分别在37℃ 水浴中和28.6℃的蒸气中解冻时玻璃化率进行研究。结果显示,在蒸气中解冻时的玻璃化率明显高于水浴中解冻;麦管玻璃化率在水浴中解冻时不稳定,在蒸气中解冻时表现稳定,玻璃化率随着玻璃化液浓度的升高有逐渐升高的趋势。对含有牙鲆胚胎的玻璃化冷冻麦管分别在37℃水浴和28.6℃蒸气中解冻,结果表明,蒸气中解冻时的玻璃化率同样明显高于水浴中解冻。 -
显微注射抗冻剂对牙鲆(Paralichthys olivaceus)胚胎毒性及冷敏感性的影响
作 者: 姜静,田永胜,翟介明,陈松林 摘 要: :在鱼类胚胎冷冻保存中,由于胚胎体积大、卵膜具多层结构,并且通透性差,传统的平衡法无法使抗冻剂足够、有效的进入胚胎内部起到保护作用。为弥补了普通平衡法的不足,此研究利用显微注射技术,将抗冻剂直接注射到鱼类胚胎卵黄囊内,提高了抗冻剂对胚胎的保护程度。此研究以牙鲆胚胎为试验材料,对显微注射抗冻剂种类、抗冻剂的注射剂量、注射浓度、以及抗冻剂注射后胚胎的冷敏感性影响进行了筛选。结果表明:牙鲆胚胎较适宜的显微注射剂量为750pl;几种抗冻剂注射牙鲆胚胎后的毒性 大小相对排列为PVP>蔗糖,DMSO>MeOH>PG,其中注射PG获得的胚胎成活率和孵化率最高,注射PVP获得的胚胎成活率和孵化率最低,而PG与MeOH组成混合抗冻剂PM毒性更低获得的胚胎成活率和孵化率更高。而任何一种抗冻剂均随着浓度的增加其毒性随之增加;另外,在冷敏感试验中,显微 注射6 mol/L的PM的胚胎胚胎,应用程序化法处理,以2~C/min的速率由室温降至一20℃ ,平衡10 min后解冻处理,发现注射PM的胚胎低温处理后成活率为(25.07~1.57)%,而6mol/的PM五步平衡法同步处理的对照组胚胎成活率为(20.88士2.84)% ,说明显微注射的胚胎冷敏感性一定降低。 -
鲈鱼胚胎的玻璃化冷冻保存
作 者: 田永胜, 陈松林,严安生,季相山,于过才 摘 要: 本文对鲈鱼 ( L ateolabrax japonicus) 胚胎进行了玻璃化冷冻保存研究 , 筛选出了浓度较低、玻璃化程度较稳定的5 种玻璃化液 , 冷冻时形成玻璃化的概率在4811 %~100 % , 在35~43 ℃ 的水浴中解冻时保持玻璃化的概率在 4414 %~6310 %; -
鲈鱼胚胎程序化冷冻保存的研究
作 者: 于过才 陈松林 孔晓瑜 田永胜 季相山 摘 要: 对鲈鱼 (Lateolabrax japonicus) 胚胎的程序降温冷冻保存进行了研究。通过稀释液的筛选实验, 选择了一种培养效果好的稀释液 DS1 液。 -
鲈鱼冷冻精子诱导半滑舌鳎胚胎发育
作 者: 田永胜 陈松林 邵长伟 刘本伟 庄志猛 摘 要: 半滑舌鳎是分布在我国黄、 渤海的特有鱼类 ,雌鱼明显具有较雄鱼生长快的特点 ,利用雌核发育方法诱导产生半滑舌鳎超雌鱼 ,通过与正常雄鱼交配 ,最终可培育生长速度快、 个体大的全雌性后代 ,对于提高养殖效益具有重要的意义。 -
花鲈冷冻精子诱导漠斑牙鲆雌核发育
作 者: 杨景峰, 陈松林, 徐亘博, 苏鹏志, 田永胜, 翟介明, 李 波 摘 要: 鱼类雌核发育工作开始于 20 世纪 50 年代,首先在淡水鱼类遗传育种的相关研究中迅速开展, 由于其母本遗传的特殊性, 雌核发育技术在基因作图与纯系的快速建立、 生产单性种群等方面有广泛的应用前景[ 1] -
花鲈冷冻精液受精鱼苗与鲜精受精鱼苗的同工酶比较分析
作 者: 陈艳翠 ,高天翔 ,陈松林 ,季相山 ,田永胜 ,邓 寒 摘 要: 采用水平凝胶电泳技术对花鲈(Lateolabrax maculatus)冷冻精液受精鱼苗与鲜精受精鱼苗进行了同工酶分析。本实验选取肌肉和肝脏两种组织,检测了IDHP、PGM、LDH、GPI、MDH、G3PDH、ME、HK、SOD、G6PD和SDH等11种酶,得到13个位点:IDHP 、PGM一1 、PGM.2 、LDH 、GPI-1 、GPI-2 、MDH 、G3PDH 、ME 、HK 、SOD 、C.6PD 和SDH 。结果显示在0.99水平,SDH 和PGM一2 位点在两种受精鱼苗中均表现为多态,多态位点比例均为0.1538;平均预期杂合度分别为0.0460和0.0407,但未观测到杂合子,观测杂合度均为0,遗传偏离指数均为一1;有效等位基因数分别为1.0720和1.0850,花鲈冷冻精液受精鱼苗与鲜精受精鱼苗的遗传相似度为0.9991,遗传距离为0.0010 -
额尔齐斯河江鳕精子冷冻保存研究
作 者: 田永胜, 姜静, 马允, 向伟, 李胜忠, 马骁, 高坚, 范镇民, 钱龙 摘 要: 齐斯河中蕴藏着丰富的冷水鱼类种质资源,江鳕(厶施fo把)是其中之一,建立其精子冷冻保存技术和精子冷冻库,对种质资源保存具有重要意义。本研究利用无机盐、葡萄糖、蔗糖、胎牛血清等配制7种精子稀释液,以5:1的比例稀释精液,激活后观察统计精子活力。结果显示,江鳕精子仅在SS、D15和CM中具有(13.33±2.89)%~(8.33±2.88)%活 -
超低温冷冻保存对七带石斑鱼胚胎酶活性的影响
作 者: 姜静, 田永胜, 翟介明, 陈松林, JIANG Jing, TIAN Yong-Sheng, ZHAI Jie-Ming, CHEN Song- Lin 摘 要: 带石斑鱼(印i唧^ek sep£e,咖sc泐凇)是一种生长速度快、低温下耐受性强具有较高经济价值的海水鱼类,在自然环境中种群数量较少,对其种质资源进行长期保存迫在眉睫。本研究以七带石斑鱼胚胎为实验材料进行种质冷冻保存,探讨超低温冷冻(一196℃)对七带石斑鱼胚胎内超氧化物歧化酶(superoxide dismutase,SOD)、肌酸激酶(creatine kinase,CK)、N。+瓜+.ATPase、乳酸脱氢酶(1actate dehydrogenase,LDH)、丙二醛(malondialdehyde,MDA)和谷胱甘肽过氧化物酶(glutathione peroxidase,GSH—PX)活性的影响,以检测抗冻剂在冷冻保存中对胚胎的影响。本研究分别测定了经过PM(24%1,2一丙二醇(PG)+16%甲醇(MeOH))、PMG(15.75%1,2一丙二醇+8.75%甲醇(MeOH)+8.75%甘油(Gly))和PMGT(15.75%1,2一丙二醇+8.75%甲醇(MeOH)+8.75%甘油(Glv)+5%海藻糖(trehalose))3种玻璃化液处理冷冻前后七带石斑鱼胚胎内6种酶活性的变化。结 -
Sperm cryopreservation of sex-reversed seven-band grouper, Epinephelus septemfasciatus
作 者: Seven-band grouper Epinephelus septemfasciatus is a protogynous hermaphrodite. The male individuals’ number is more less than the female one. Thus, it is necessary for artificial reproduction and crossbreeding to research the sperm cryopreservation of sex-reversed seven-band grouper. 摘 要: Seven-band grouper Epinephelus septemfasciatus is a protogynous hermaphrodite. The male individuals’ number is more less than the female one. Thus, it is necessary for artificial reproduction and crossbreeding to research the sperm cryopreservation of sex-reversed seven-band grouper. -
Sperm cryopreservation of sex-reversed seven-band grouper, Epinephelus septemfasciatus
作 者: Yongsheng Tiana,∗, Wenshan Qia,b, Jing Jianga,b, Na Wanga, Di Wanga,c, Jieming Zhaid, Chao Chena, Songlin Chena,∗ 摘 要: Seven-band grouper Epinephelus septemfasciatus is a protogynous hermaphrodite. The male individuals’ number is more less than the female one. Thus, it is necessary for artificial reproduction and crossbreeding to research the sperm cryopreservation of sex-reversed seven-band grouper. -
Long-term cryopreservation of sperm from Mandarin fish Siniperca chuatsi
作 者: Shuyan Dinga, Jiachun Gea,∗, Chen Haoa, Mingsheng Zhanga, Weihui Yana, ZhiqiangXua, Jianlin Pana, Songlin Chenb, Yongsheng Tianb, YahongHuang 摘 要: In order to develop cryopreservation techniques for long-termpre-serving the sperm of Mandarin fish Siniperca chuatsi, we examined the effects of various extender and cryopreservation on post-thawmotility. -
Effects of cryopreservation on the survival rate of the seven-band grouper (Epinephelus septemfasciatus) embryos
作 者: Yongsheng Tian a, *, 1, Jing Jiang a, b, 1, Lini Song a, b, Zhangfan Chen a, Jieming Zhai c, Jiangchun Liu c, Na Wang a, Songlin Chen a, * 摘 要: The effects of cryopreservation and the vitrification solution on the embryo hatchability of the seven- band grouper Epinephelus septemfasciatus were evaluated in this study. Six small molecule cryoprotec-tants (PG, MeOH, Gly, DMF, DMSO and EG) and four macromolecular cryoprotectants (glucose, fructose, sucrose and trehalose) were used to determine the embryo toxicity levels. -
Cryopreservation of sperm from turbot (Scophthalmus maximus) and application to large-scale fertilization
作 者: Song-Lin Chena,*, Xiang-Shan Jia,b, Guo-Cai Yua, Yong-Sheng Tiana, Zhen-Xia Shaa 摘 要: This paper reports on cryopreservation of sperm from turbot (Scophthalmus maximus). The effects of various extenders, cryoprotectants and sperm–egg insemination ratios on motility score and/or fertilization capacity of post-thaw turbot spermatozoa were examined to optimize cryopreservation procedures. -
Cryopreservation of spotted halibut (Verasper variegatus) sperm
作 者: Y.S. Tian a, S.L. Chen a,⁎, X.S. Ji b, J.M. Zhai c, L.J. Sun c, C. Chen a, P.Z. Su a 摘 要: Spotted halibut (Verasper variegatus) is now regarded as being endangered in China because of its continuous decline in the last three decades. Preservation and protection of its genetic resources were demanded in order to protect this rare fish species -
Cryopreservation of sea perch (Lateolabrax japonicus) spermatozoa and feasibility for production-scale fertilization
作 者: X.S. Jia, S.L. Chena,b,*, Y.S. Tiana, G.C. Yua, Z.X. Shaa, M.Y. Xua, S.C. Zhangc 摘 要: With the objective of germplasm conservation and cryobank construction, a method for cryopreserving sea perch (Lateolabrax japonicus, Cuvier) semen in 1.8-ml cryovials was developed. The effects of various extenders, cryoprotectants, volume of diluted semen on the motility score of post-thaw spermatozoa were examined. -
云纹石斑鱼心脏细胞系的建立与鉴定
作 者: 刘肖峰 陈松林 沙珍霞 吴娅红 于洋 王娜 修旺珊2 摘 要: 鱼类细胞系在鱼类病毒学及功能基因组学方面发挥着重要作用。云纹石斑鱼(Epinephelusmoara) 是一种具有重要养殖价值的海水鱼类, 尚未见细胞系建立的报道。本研究以云纹石斑鱼的心脏组织为材料, 建立了云纹石斑鱼心脏细胞系 -
半滑舌鳎脾脏"卵巢组织细胞离体培养的初步研究
作 者: 郑媛 陈松林 摘 要: 44半滑舌鳎#-$)*#1*33233,:"1&,F"3$ 属鲽形目#60>=?/C>23;Q/?9>A$ " 舌鳎科#-@C/E0/AA;B:>$ " 舌鳎属#-@C/E0/AA=A$ " 俗称牛舌头% 鳎目% 鳎米" 是一种暖温性近海大型底层鱼类"终年生活栖息在我国近海海区&1'"具广温%广盐和适应多变的环境条件的特点( 半滑舌鳎市场价值高"但自然资源量少"是我国新开发的重要海水养殖鱼类