锯缘青蟹不同器官组织中总抗氧化能力和SOD活性的比较研究

采用生化测定的方法,对锯缘青蟹的鳃、肝胰腺和肌肉等不同器官组织中的总抗氧化能力和超氧化物歧化酶活性进行了测定.结果表明,不同器官组织中的总抗氧化能力和超氧化物歧化酶活性各异.总抗氧化能力由强至弱依次为肝胰腺>肌肉>鳃,分别为71.16±17.75U/mg蛋白,28.55±4.45U/mg蛋白,19.82±3.20U/mg蛋白.而超氧化物歧化酶活性由强至弱则依次为肝胰腺>鳃>肌肉,分别为38.62±5.24 NU/mg蛋白,27.08±4.79 NU/mg蛋白,6.33±1.54 NU/mg蛋白.统计分析显示,各器官组织间的总抗氧化能力和超氧化物歧化酶活性有显著性差异(p<0.01),这与不同器官组织的生理功能是密切相关的.     

金钱鱼鳃细胞系的建立及其生长特性的初步研究

鳃是鱼类渗透压调节的主要器官, 鳃细胞的培养技术的建立可以为鱼类渗透压调节机理研究提供重要的实验手段和研究方法。金钱鱼(Scatophagus argus)作为一种广盐性鱼类, 是研究渗透压调节机制的理想动物模型。本研究采用胰蛋白酶消化法进行了金钱鱼鳃细胞的体外培养, 确定该类细胞原代和传代培养的最优条件, 分析了其生长特性。结果表明, 金钱鱼鳃细胞系在28℃, 含有20%胎牛血清(fetal bovine serum, FBS)的L-15培养基中生长最佳, 2~4d即可传代, 传代后可稳定培养, 命名为SG。在传代培养过程中, 对其增殖情况进行分析, 发现SG细胞群体倍增时间为40.8h。正常传代的SG细胞冻存、复苏后, 经台盼蓝染色测得细胞存活率约为87%。鳃细胞直接同水体环境接触, 具有高效的渗透压调节能力。环境盐度变化时, 鳃细胞启动渗透压胁迫应答机制, 维持内环境稳态。本研究对SG细胞进行渗透压刺激后, 检测其增殖情况并观察形态变化。分析表明, SG细胞在分别为150和600mOsmol·kg ^-1的低渗和高渗胁迫后均可增殖, 且低渗胁迫后的增殖速度是高渗胁迫的1.5倍。渗透压胁迫后SG细胞的形态观察结果显示, 低渗培养基胁迫后细胞体积发生膨胀而高渗条件下细胞体积发生皱缩。由此推断SG细胞具有较强渗透压耐受性, 且低渗耐受能力强于高渗。SG细胞系的建立为金钱鱼渗透压应答机制和相关基因功能的研究提供了基础实验材料。     

栉孔扇贝鳃、唇瓣和口唇一氧化氮合酶活性

采用组织化学、免疫组织化学和分光光度技术对栉孔扇贝(Chlamys farreri)鳃、唇瓣和口唇内的一氧化氮合酶(EC EC1.14.13.39;NOS)进行了研究。组织化学显示,鳃轴结缔组织内血细胞、血管内皮细胞、神经细胞、神经纤维呈NOS阳性;鳃丝的部分上皮细胞及血细胞呈NOS强阳性;唇瓣内部分上皮细胞、血细胞、神经细胞和神经纤维均呈NOS阳性;口唇有大量的神经纤维呈极强的NOS阳性。免疫组化显示,鳃和唇瓣血细胞、神经细胞、神经纤维和部分上皮细胞呈iNOS阳性。口唇内血细胞、神经细胞、神经纤维和部分上皮细胞呈nNOS和eNOS阳性,而呈iNOS强阳性。生化测定表明,tNOS活力和NO含量均为鳃轴中最高,鳃丝中次之,唇瓣中较低,口唇中最低,鳃丝内的NOS以cNOS为主;鳃轴和口唇内的cNOS活力和iNOS活力相近;唇瓣内的NOS则以iNOS为主。     

盐度突变对半滑舌鳎血浆渗透压和鳃丝Na+/K+-ATP酶活性的影响

研究了半滑舌鳎(Cynoglossus semilaevis)由盐度30突变至0、1O、20、35和40盐度后血浆渗透压和鳃丝Na+/K+-ATP酶活性的变化.结果表明,盐度对半滑舌鳎血液渗透压和鳃丝Na+/K+-ATP酶活性均有显著影响(P＜0.05).盐度突变后,各处理组的血液渗透压和鳃丝Na+/K+-ATP酶活性...     

Thraustochytrium caudivorum——一种香港牡蛎新型原生生物寄生虫

破囊壶菌(Thraustochytrids)是一类具有分解作用的海洋异养原生生物。研究表明破囊壶菌可对软体动物、海洋扁虫类造成消耗性疾病。本研究从发病的牡蛎鳃组织中分离出一种未在牡蛎中记录过的破囊壶菌寄生虫,经形态学及分子生物学分析后,鉴定为一种破囊壶菌寄生虫Thraustochytriu caudivorum。利用倒置显微镜和激光共聚焦超高分辨系统观察到T.caudivorum的生活史可分为游动孢子、营养细胞、孢子囊和变形虫四个阶段。游动孢子沉降成营养细胞,营养细胞再通过二分裂或形成孢子囊的方式释放不动的和可游动的孢子。破囊壶菌的形态大小变化迥异,本研究利用流式细胞仪和共聚焦系统探究了不同盐度对细胞大小的影响。结果表明,随盐度增大,体积小的细胞占比增多;在盐度为15 ppt时,细胞增殖缓慢,体积变大;而在盐度为20和25 ppt时,细胞体积变小,增殖速率加快。25 ppt盐度条件下,游动孢子数目最多。本研究首次从香港牡蛎鳃组织中分离出T. caudivorum寄生虫,并确立了其与破囊壶菌其他属之间的系统发育关系。     

重金属离子对中华绒螯蟹肝胰脏和鳃丝SOD,CAT活力的影响

研究了 3种重金属离子 (Cu2 ,Zn2 ,Cd2 )对中华绒螯蟹肝胰脏和鳃丝 SOD,CAT活力的影响。实验结果表明 ,肝胰脏和鳃丝 SOD,CAT活力在 3种重金属离子作用下分别随取样时间变化显著 (P<0 .0 5 ) ,对 2种抗氧化酶活力的抑制均为肝胰脏 <鳃丝。 3种重金属离子在实验时间内对肝胰脏 2种抗氧化酶活力的影响呈峰值变化 ,低浓度组表现为促进作用 ,高浓度组在 96 h时为抑制作用 ;低浓度组在 72 h内能提高鳃丝 2种抗氧化酶活力 ,随后下降 ,至 96 h时表现为抑制作用 ,而高浓度组一直为明显的抑制作用 ,2种抗氧化酶活力表现出明显的时间、剂量效应关系。因此 ,肝胰脏和鳃丝SOD,CAT2种抗氧化酶可作为中华绒螯蟹毒理学的评价指标。     

Spatial variability in branchial basket meristics and morphology of southern African sardine Sardinops sagax

We examined spatial variability in meristic and morphological characteristics of the branchial basket of sardine Sardinops sagax collected from four geographical regions around the southern African coast, namely Namibia and the South African west, south and east coasts. Our analysis tested the hypothesis of three putative sardine stocks off South Africa, one in each of the three geographical regions. We therefore collected fish data from Namibia to compare with South Africa, because sardine from the two countries are considered to be separate stocks. Morphometric measurements (gill arch length and gill raker spacing) and meristic data (number of gill rakers) were collected from the left side of the first gill arch from a total of 377 sardine, approximately equally divided between the regions. A multivariate general linear model with caudal length as covariate was used to assess differences among fish from the four regions and significant differences were observed, although not always consistently across all fish size classes. Small South Coast sardine had shorter gill arches than small West Coast sardine, but adults had gill arches of similar length, longer than those from Namibia and the East Coast. Small sardine from the South Coast had fewer gill rakers than small sardine from the West Coast, but larger fish had similar numbers of gill rakers, significantly more than sardine from Namibia and the East Coast. Sardine from the West and South coasts had similar gill raker spacings, which were smaller than those of fish from the East Coast and Namibia. Despite spatially and particularly temporally unbalanced sampling, we consider that these differences provide evidence of spatial variation in Benguela sardine phenotype and that it would support the hypothesis of discrete sardine stocks off Namibia and South Africa. The results are consistent with the hypothesis of three sardine stocks within the southern Benguela.     

Identification and characterization of the Vibrio anguillarum prtV gene encoding a new metalloprotease

We cloned and sequenced a prtV-like gene from Vibrio anguillarum M3 strain.This prtV gene encodes a putative protein of 918 amino acids,and is highly homologous to the V.cholerae prtV gene.We found that a prtV insertion mutant strain displayed lower gelatinase activity on gelatin agar,lower protease activity against azocasein,and lower activity for four glycosidases.This prtV mutant strain also had increased activity for two esterases in its extracellular products,as analyzed by the API ZYM system.In additi...     

Effects of waterborne Fe(II) on juvenile turbot Scophthalmus maximus: analysis of respiratory rate, hematology and gill histology

The concentration of Fe(Ⅱ) is high in some groundwater supplies used in turbot culture,and the toxicity of waterborne Fe(Ⅱ) is unknown.We investigated the stress responses of juvenile turbot,Scophthalmus maximus,exposed to Fe(Ⅱ) of different concentrations (0.01,0.05,0.1,0.5,1,and 2 mg/L) for 1,7,14,and 28 d,under the same ambient conditions of other parameters.Changes in respiratory rate,hematological parameters,and gill structure were determined.The results show that waterborne Fe(Ⅱ) did not cause severe hematological perturbation to turbot.A low-medium Fe(Ⅱ) concentration (lower than 0.1 mg/L) could boost the respiratory rate,and caused no or very limited damage to fish.A high Fe(Ⅱ) concentration (0.1 mg/L or higher),however,caused gill damage,such as vacuoles in branchial lamellae,epithelial necrosis,and hypertrophy of epithelial cells,and even death after extended exposure time.Therefore,excess waterborne Fe(Ⅱ) and long-term exposure to Fe(Ⅱ) could be responsible for poor growth and high mortality of turbot in culture.The concentration of waterborne Fe(Ⅱ) in turbot culture should be kept below 0.1 mg/L.