温度对尼罗罗非鱼急性耐盐性的影响

采用两因素交叉分组的方法,初步研究了温度对尼罗罗非鱼急性耐盐性的影响。结果表明:温度与盐度的交互作用非常明显(P<0.05),在温度为25、28、31℃时,尼罗罗非鱼的96h半致死盐度(MSL-96)分别为26.2、22.5、16.6,三者之间差异显著(P<0.05),表现出随温度升高而MSL-96降低的趋势。不同盐度和温度水平的平均成活时间(MST)均差异显著(P<0.05),同时也表现出随温度和盐度升高而MST降低的趋势;在同一温度下各盐度水平的50%死亡时间(ST50)差异显著(P<0.05)。     

不同培养液添加物对尼罗罗非鱼精原干细胞生长增殖的影响

【目的】探讨不同血清和细胞因子等培养液添加物对尼罗罗非鱼(Oreochromisniloticus)精原干细胞(Spermatogonial stem cells,SSCs)生长增殖的影响。【方法】无菌取发育第Ⅲ期的尼罗罗非鱼精巢组织,用酶消化法结合差速贴壁法获得尼罗罗非鱼SSCs,在有支持细胞饲养层条件下,L-15培养液中分别添加体积分数2%、5%、10%新生牛血清(New bovine serum,NBS),体积分数1%、2%、3%罗非鱼血清以及5、10、15 ng/mL白血病抑制因子(Leukemia inhibitor factor,LIF),比较不同添加物对尼罗罗非鱼SSCs生长增殖的影响。【结果】在支持细胞饲养层上,SSCs分裂指数明显高于无饲养层的SSCs,支持细胞饲养层明显促进精原干细胞分裂增殖(P 0.01);与2%、10%NBS组相比,添加体积分数5%NBS可显著促进精原干细胞增殖(P 0.01);添加体积分数1%、2%、3%尼罗罗非鱼血清对SSCs无显著的促增殖作用(P 0.05);添加5、10 ng/mL LIF可促进SSCs增殖,添加10 ng/mL LIF效果更佳,添加15 ng/mL LIF则抑制SSCs增殖。【结论】使用尼罗罗非鱼精巢支持细胞作饲养层,用添加体积分数5%NBS及10 ng/mL LIF的L-15培养液培养,有助于促进尼罗罗非鱼精原干细胞生长增殖。     

温度、盐度、pH和体质量对尼罗罗非鱼幼鱼窒息点的影响

利用实验生态学的方法研究不同温度(15、20、25、30和35℃)、盐度(0、5、10、15和20)、pH(5、6、7、8和9)和体质量(1.93±0.08、3.43±0.12、7.95±0.34、11.74±0.41和15.84±0.61 g)对尼罗罗非鱼幼鱼窒息点的影响,并建立温度、盐度、pH、体质量与罗非鱼窒息点间的回归方程。结果表明:温度t、盐度S、pH和体质量m对尼罗罗非鱼幼鱼窒息点有显著影响(P<0.05)。当温度、盐度、pH和体质量分别为25.28℃、3.6、8.06、6.3 g时,窒息点(DAP)最低,回归方程为DAP=0.000 9 t 2-0.045 5 t+0.726 6(R2=0.952 1),DAP=-0.000 1 S 3+0.004 2 S 2-0.023 9 S+0.111 9(R2=0.922 8),DAP=0.009 3 pH2-0.149 9 pH+0.7174(R2=0.976 8);DAP=-0.000 3 m3+0.011 1 m2-0.104 3 m+0.362 8(R2=0.948 4)。在溶氧不足情况下,温度25.28℃、盐度3.6、pH 8.06的环境条件有助于提高幼鱼耐低氧能力。     

盐度对莫荷罗非鱼幼鱼呼吸和氨氮排泄的影响

将莫荷罗非鱼(Oreochromis mossambicus×O.urolepis hornorum)幼鱼养殖水体盐度从淡水渐变至12、18、24、30(每天提升6)等4个目标盐度,利用间歇式呼吸仪分析幼鱼在目标盐度下5、10、15、20 d时的呼吸和氨氮排泄的变化。结果表明,在实验前期,莫荷罗非鱼幼鱼耗氧率、排氨率随盐度的升高而增加,各盐度处理组均在5 d时升到最大值,之后开始下降,下降时间、幅度与外界盐度有关。盐度12、18组幼鱼耗氧率10 d时下降至对照组水平,20 d时低于对照组水平,差异有统计学意义(P<0.05);盐度24、30组幼鱼耗氧率则在15 d时下降至对照组水平并维持稳定;幼鱼排氨率随实验时间的延长,各盐度处理组幼鱼排氨率呈先上后下降的变化,盐度24、30组幼鱼排氨率在15 d下降并维持在对照组水平,盐度12、18组幼鱼排氨率在15 d时下降至对照组水平并于20 d时低于其他盐度组,差异有统计学意义(P<0.05)。莫荷罗非鱼幼鱼适应新盐度环境后,盐度18组的耗氧率、排氨率最低,与盐度12组差异无统计学意义(P>0.05),与其他盐度处理组有差异有统计学意义(P<0.05),推测其等渗点在盐度12~18之间。不同盐度条件下,氧氮比值介于18.810~24.216间,氨熵介于0.083~0.108间,表明莫荷罗非鱼幼鱼主要依靠蛋白质和脂肪氧化供能。     

尼罗罗非鱼淋巴毒素α基因克隆与表达分析

【目的】克隆尼罗罗非鱼(Oreochromis niloticus)淋巴毒素α基因(lymphotoxin alpha, LTα),分析该基因的组织分布,探讨该基因在抗菌抗病毒免疫过程中的重要作用。【方法】通过RACE技术获得尼罗罗非鱼淋巴毒素α基因(On-LTα)的cDNA全长,通过生物信息学分析其序列结构特征;利用实时荧光定量PCR检测基因的组织分布特征,以及健康尼罗罗非鱼感染无乳链球菌后该基因的表达变化,并进一步分离出尼罗罗非鱼非特异性毒性细胞(NCC),检测脂多糖(LPS)和聚肌胞苷酸(Poly I:C)刺激后,On-LTα在NCC中的表达变化。【结果】On-LTα基因序列全长为2699 bp,包含705 bp开放阅读框(ORF),编码234个氨基酸(GenBank登录号:MK770358)。该蛋白属于跨膜蛋白,具有肿瘤坏死因子(TNF)家族保守结构域。实时荧光定量结果表明,On-LTα在健康尼罗罗非鱼11种组织中均有表达,在脾脏中的表达量最高;无乳链球菌刺激后,该基因在脾脏中的表达量显著升高;LPS与PolyI:C刺激后,On-LTα在NCC中表达量也显著上调。【结论】On-LTα参与了尼罗罗非鱼抗菌抗病毒的免疫应答过程。     

不同温度下罗非鱼人工感染海豚链球菌后血液生化指标及组织病理

在28℃、33℃两种温度下,用海豚链球菌人工感染尼罗罗非鱼后,测定尼罗罗非鱼血液生化指标。结果显示:两个温度水平病鱼的血清总蛋白(TP)随时间延长都呈现升高趋势;谷丙转氨酶(ALT)、谷草转氨酶(AST)活性随时间延长先升高后降低再升高,33℃实验组ALT、AST活性比28℃实验组达峰值时间短,并显著高于对照组(p<0.05);尿素、钾(K+)含量随时间延长逐渐升高后降低,钠(Na+)含量先升高后降低再升高,肌酐含量呈上升趋势;血清碱性磷酸酶(AKP)活性28℃实验组呈先升高后降低趋势,33℃实验组呈升高趋势;超氧化歧化酶(SOD)活性随时间延长先升高后降低再升高,33℃实验组各个时间点基本高于28℃实验组;球蛋白(GLO)随感染后时间延长呈现升高趋势,至120 h,显著高于对照组(p<0.05),但温度组间各时间点没有显著差异(p>0.05)。组织病理学观察发现,33℃实验组肝细胞、肾小管上皮细胞水肿变性;28℃实验组肝细胞出现嗜酸性小体;肾小管上皮细胞出现嗜酸性的红色小体,颗粒变性。表明感染海豚链球菌的尼罗罗非鱼肝脏、肾脏均有损伤,造成功能障碍,高温时损伤更严重。     

MS-222和苯唑卡因麻醉后罗非鱼血清电解质变化及其与红细胞数的相关性

分别用3个浓度梯度的MS-222和苯唑卡因对(6.05±1.71)g的尼罗罗非鱼(Oreochromis niloticus)进行6 h浸泡式麻醉,抽取尾部静脉血,研究血清中K~+、Na~+、Cl~-、Ca~(2+)等4种无机离子水平变化以其与红细胞数的相关性。结果表明:1)随MS-222质量浓度的升高,血清中K+浓度呈持续下降趋势,Na+浓度保持相对稳定,Cl-浓度逐步升高,Ca~(2+)浓度呈现出峰谷式波动,各离子浓度变化与红细胞数之间相关系数绝对值大小依次为Cl-(0.911 6)、K+(-0.551 9)、Na~+(0.337 5)、Ca~(2+)(0.208 9),红细胞数升高的同时发生了Cl-外流;2)随苯唑卡因质量浓度的升高,血清中K~+浓度呈峰谷式波动,Na+浓度亦保持相对稳定,Cl-浓度呈现小幅波动,Ca~(2+)浓度基本呈下降趋势,离子浓度与红细胞数之间相关系数绝对值大小依次为Ca~(2+)(0.970 1)、Na~+(0.315 2)、Cl~-(0.265 3)、K~+(-0.049 9),红细胞数降低的同时发生了Ca~(2+)内流;3)两种药物处理后,离子之间的相关性大小次序基本一致,均为K~+-Na~+最高,其次是K~+-Cl~-和Na~+-Cl~-。     

奥利亚罗非鱼微卫星位点的筛选

从GenBank上选取40对尼罗罗非鱼(Oreochromis niloticus)的微卫星引物,分别在奥利亚罗非鱼(Oreo-chromis aureus)(83系)、红色奥利亚罗非鱼、奥利亚罗非鱼(02系)和尼罗罗非鱼的基因组上进行扩增。40对引物中有37对(92.5%)可进行有效扩增,32对引物(80%)可检测到个体间等位基因的多态性。其中引物UNH168与奥利亚罗非鱼的性别相关,在雌性个体中可扩增出二条大小不同的特异带(分别为135和171 bp),在雄性个体中则只有一条(171 bp)。将特异条带回收、克隆并测序,结果显示雌雄个体中171 bp条带的序列完全相同,包括103bp的侧翼序列和34个CA重复,在雌性个体中获得的135 bp条带则只有16个CA重复。引物UNH846、UNH860和UNH995可鉴别奥利亚罗非鱼和尼罗罗非鱼,UNH890可鉴别出红色奥利亚罗非鱼。可见,大部分的尼罗罗非鱼微卫星位点存在于奥利亚罗非鱼中。     

运用RAPD技术分析3种罗非鱼的遗传多样性

应用RAPD技术,从7个系列140种随机引物中筛选出15个,对尼罗罗非鱼(Oreochromis niloticus)、奥利亚罗非鱼(O.aureus)和星洲银罗非鱼(O.sp.)进行遗传多样性分析。结果表明,尼罗罗非鱼、奥利亚罗非鱼、星洲银罗非鱼的多态位点比例(P)分别为54.7%、44.4%和56.0%,种内个体间遗传距离分别为0.145 0、0.115 6和0.141 3,种内遗传相似系数分别为0.855 0、0.884 6和0.858 7;种间遗传距离的聚类分析表明,星洲银罗非鱼与尼罗罗非鱼亲缘关系较近,一定程度上验证了星洲银罗非鱼的分类地位。     

尼罗罗非鱼干扰素调节因子4基因的克隆与表达

【目的】探究干扰素调节因子(IRF)在尼罗罗非鱼(Oreochromisniloticus)免疫中的作用。【方法】从尼罗罗非鱼中鉴定IRF4同源物,命名为OnIRF4。使用生物信息学分析软件分析OnIRF4的氨基酸序列。对健康尼罗罗非鱼腹腔注射浓度为3×10~7 CFU/mL的无乳链球菌200μL,用实时荧光定量PCR法分析OnIRF4在健康鱼中的表达模式及其对细菌感染的生物学效应。【结果与结论】OnIRF4序列的开放阅读框为1 350 bp,编码449个氨基酸。OnIRF4与其他物种的IRF4蛋白的同源性为52%～98%。推导的OnIRF4肽链具有干扰素因子典型的DNA结合结构域(DBD)、干扰素相联结构域(IAD)。亚细胞定位结果表明,OnIRF4主要定位在细胞核。表达分析表明,OnIRF4在健康尼罗罗非鱼皮肤、头肾、鳃、脑、肠、肌肉、肝、胸腺和脾中均有分布。在无乳链球菌(Streptococcus agalactiae)感染后,OnIRF4在肝脏、脾脏、头肾、脑中的表达显著上调(P0.05),表明OnIRF4参与了尼罗罗非鱼对细菌感染的免疫反应。     

盐度对威氏海链藻生长和生化指标的影响

【目的】研究威氏海链藻在不同盐度条件下的生长和生化组分变化。【方法】设置盐度梯度为5、15、25、35、45,采用血细胞计数板计数法、热乙醇提取法、尼罗红染色法、苯酚硫酸法和考马斯亮蓝法,分别测定威氏海链藻在不同盐度下的生物量、叶绿素a、类胡萝卜素、中性脂质、总糖和蛋白质含量。【结果】盐度对威氏海链藻细胞密度影响显著(P<0.05),当盐度25和盐度35时,培养8 d细胞密度最高,均为2.0×10⁶ mL^-1;盐度对威氏海链藻叶绿素a和类胡萝卜素含量有显著影响(P<0.05),当盐度15时,细胞叶绿素a和类胡萝卜素含量均最高,分别为3.4814μg/mL(4 d)、2.3549μg/mL(8 d)。盐度对威氏海链藻可溶性总糖含量有显著影响(P<0.05),当盐度15时,8 d时可溶性总糖积累含量最高,为123μg/mL。当盐度35时,6 d时可溶性蛋白积累含量最高,为142.9μg/mL,且显著高于其他实验组(P<0.05)。盐度45时,8 d时中性脂含量最高,显著高于其他实验组(P<0.05)。【结论】盐度为15~35时,威氏海链藻生长速度较快,生化组成较为稳定,盐度15时最有利于藻细胞叶绿素a和类胡萝卜素、总糖的合成;盐度25和盐度35时,藻细胞平均增长速度较快,盐度35时最有利于藻细胞可溶性蛋白积累。     

Salinity tolerance and growth response of juvenile <Emphasis Type="Italic">Oreochromis mossambicus</Emphasis> at different salinity levels

1　Introduction　Theuseofnaturalresources ,includingthosebeingofseeminglymarginalvalue ,isanimportanthumanactivitydesignedtoincreasefoodproductionandin come .Inagriculture ,soilsmaybetoosalinetosup portprofitablecrophusbandry ,yetsuchsoilmaybeusedalternat…     

盐度对菊黄东方鲀受精卵孵化和仔鱼生长的影响

比较在0、5、10、15、20、25、30(自然海水)、35、40、45、50等11个盐度梯度下,菊黄东方鲀(Takiugu flavidus)的受精卵发育和仔鱼生长情况,测定仔鱼的不投饵存活系数(SAI).结果表明:1)菊黄东方鲀受精卵在盐度0~50范围内均可孵化,最适盐度20~25.盐度(x)与孵化时间(y)呈正相关关系,表达式为:y=0.345 x+141.1,R2=0.956.2)实验盐度范围内,菊黄东方鲀仔鱼SAI值为1.55~30.10.3)在盐度5~45条件下,仔鱼皆可存活,存活率为3%~27%.其中,在10~30盐度下存活率较高,为20%~27%.经15 d的培育,仔鱼平均全长为6.28~7.34 mm.低盐度对菊黄东方鲀仔鱼生长更有利,在15~20盐度条件下,仔鱼生长较好,最终平均全长为7.30~7.34 mm.4)根据SAI值和生长状况,仔鱼生长和存活的适宜生长盐度为10~30,最适盐度范围为15~20.     

Effects of salinity on egg and fecal pellet production,development and survival,adult sex ratio and total life span in the calanoid copepod,A cartia tonsa:a laboratory study

The ef fects of salinity on the copepod, A cartia tonsa in terms of daily egg production rate(EPR), hatching success, fecal pellet production rate(FPR), naupliar development time and survival, sex ratio, and total life span were determined in laboratory conditions through three experiments. In experiment 1, EPR, hatching success, and FPR of individual females were monitored at salinities of 13, 20, 35 and 45 during short-periods(seven consecutive days). Results show EPR was aff ected by salinity with the highest outputs recorded at 20 and 35, respectively, which were considerably higher than those at 13 and 45. Mean FPR was also higher in 35 and 20. In experiment 2, the same parameters were evaluated over total life span of females(long-term study). The best EPR and FPR were observed in 35, which was statistically higher than at 13 and 20. In experiment 3, survival rates of early nauplii until adult stage were lowest at a salinity of 13. The development time increased with increasing of salinity. Female percentage clearly decreased with increasing salinity. Higher female percentages(56.7% and 52.2%, respectively) were signifi cantly observed at two salinities of 13 and 20 compared to that at 35(25%). Total longevity of females was not af fected by salinity increment. Based on our results, for mass culture we recommend that a salinity of 35 be adopted due to higher reproductive performances, better feeding, and faster development of A. tonsa.     

Effects of salinity on egg and fecal pellet production,development and survival,adult sex ratio and total life span in the calanoid copepod, <Emphasis Type="Italic">Acartia tonsa</Emphasis>: a laboratory study

The effects of salinity on the copepod, Acartia tonsa in terms of daily egg production rate (EPR), hatching success, fecal pellet production rate (FPR), naupliar development time and survival, sex ratio, and total life span were determined in laboratory conditions through three experiments. In experiment 1, EPR, hatching success, and FPR of individual females were monitored at salinities of 13, 20, 35 and 45 during short-periods (seven consecutive days). Results show EPR was affected by salinity with the highest outputs recorded at 20 and 35, respectively, which were considerably higher than those at 13 and 45. Mean FPR was also higher in 35 and 20. In experiment 2, the same parameters were evaluated over total life span of females (long-term study). The best EPR and FPR were observed in 35, which was statistically higher than at 13 and 20. In experiment 3, survival rates of early nauplii until adult stage were lowest at a salinity of 13. The development time increased with increasing of salinity. Female percentage clearly decreased with increasing salinity. Higher female percentages (56.7% and 52.2%, respectively) were significantly observed at two salinities of 13 and 20 compared to that at 35 (25%). Total longevity of females was not affected by salinity increment. Based on our results, for mass culture we recommend that a salinity of 35 be adopted due to higher reproductive performances, better feeding, and faster development of A. tonsa.     

Effect of salinity on growth and first sexual maturity of Exopalaemon carinicauda (Holthuis, 1950)

To determine the optimal salinity for growth and first sexual maturity of ExopaIaemon carinicauda, the effects of salinity on growth and reproductive performance of early juvenile prawns of E. carinicauda were evaluated under laboratory conditions. Postlarvae from the same female broodstock were reared at six salinity levels （5, 10, 15, 20, 25, and 30） for 12 weeks. The specific growth rate （SGR） and survival rate （SR） under different salinity levels in the first 6 weeks were calculated and compared. SGR was significantly influenced by salinity. Prawns reared in salinity of 10 grew significantly faster （P〈0.05） than those reared in salinities of 5, 20, 25, and 30. However, ANOVA confirmed that there was no significant effect among the six salinity levels on SR. For the next 6 weeks, the body length （BL）, body weight （BW） at the first sexual maturity, and the age at median sexual maturity （As0） of females were measured and compared. Female prawns reared at salinity of 10 presented significantly shorter A50, but no significant differences among the six salinity levels for BL and BW at the first sexual maturity were observed. Based on the above information, the optimal salinity for growth and first sexual maturity of juvenile E. carinicauda is approximately 10.     

Effects of salinity on embryonic development,survival, and growth of Crassostrea hongkongensis

This study examined the effects of salinity on embryonic development, survival, and growth of the Hong Kong oyster Crassostrea hongkongensis. The embryos, larvae, and juveniles of C. hongkongensis were held in artificial seawater at three different salinities (low, 15; medium, 23; and high, 30) to determine the optimum hatchery and nursery conditions for mass production of the seeds. Results showed that the percentage production of straight-hinged larvae from fertilized eggs was significantly lower at the high salinity than at the low- and medium-salinities (P < 0.05). The survival rates of larvae and juveniles differed significantly among the three salinity trials, with the highest survival rate observed at the low salinity (P < 0.05). The shell height of larvae was significantly larger at the low salinity than at the high and medium salinities from days 9 to 15 (P < 0.05), whereas that of juveniles was significantly larger at the low salinity than at the high and medium salinities on day 70 (P < 0.05). These results indicate that the larvae and juveniles of C. hongkongensis are tolerant to a wide range of salinities (15 to 30), but show better growth and survival at relatively low salinities. Thus, it is recommended to use relatively low salinities in hatchery and nursery systems for improved yields of C. hongkongensis.     

墨西哥湾扇贝稚贝盐度适应性的研究

在室内控制条件下,观察不同海水盐度对墨西哥湾扇贝(Argopecten irradians concentricusSay)稚贝存活及生长的影响,并将168 h 50%死亡的盐度作为适宜生存临界盐度,将增长率为最佳增长率30%时所对应的盐度作为适宜生长临界盐度。结果表明:在稚贝壳高4.38 mm±0.31 mm,水温25.4~27.8℃,pH值7.8~8.2下,墨西哥湾扇贝稚贝适宜生存盐度为23.6~35.7,最适生存盐度为26.0~31.3;适宜生长盐度为23.9~35.8,最适生长盐度为28.7~31.3。在盐度变化超出最适范围时,`墨西哥湾扇贝稚贝对低盐更为敏感,高盐比低盐对稚贝存活、生长、增重的影响小。当盐度为28.7时,墨西哥湾扇贝稚贝具有最高的相对存活率(100%)、壳高日增长率(0.116 2 mm.d-1)及体重日增长率(1.95 mg.d-1)。当盐度在最适范围之外升高或降低,稚贝的存活率、壳高及体重日增长率也随着降低。