Ultrastructural changes in fish gills as biomarker to assess small stream pollution

In order to verify the principal suitability of gill ultrastructure as a biomarker, semi-field studies with two endigoneous fish, trout (Salmo trutta f. fario) and loach (Barbatula barbatula), were performed. The fish were exposed in flow-through systems to one heavily polluted (K?rsch) and one lightly polluted small stream (Kr?henbach) in South-West Germany. Ultrastructural responses in gills were correlated with limnological and chemical data recorded over a 2 year period in each stream. After 8 weeks of exposure to the heavily polluted stream, fish showed ultrastructural changes in the gills, such as cell proliferation, dilation of the endoplasmic reticulum, hyperplasia, hypersecretion, and epithelial lifting in chloride, epithelial, and mucus cells. The results of the study demonstrate that ultrastructural reactions in the gills of fish kept under semi-field conditions are potentially useful biomarkers indicating small stream pollution.     

温度、pH和盐度对克氏原螯虾鳃 Na+-K+-ATPase活性的影响

采用钼蓝法测定克氏原螯虾(Procambarus clarkii)鳃Na~ -K~ -ATPase活性,探讨温度、pH、盐度3个环境因子在环境驯化和突变状态下对Na -K -ATPase活性的影响。实验结果表明,Na~ -K~ -ATPase的活性与环境因子密切相关。酶活性在温度、盐度驯化实验中都表现为正相关关系,不同pH驯化中则表现为中性pH活性最高。在突变状况下表现出明显的应激性,应激响应在2～8h之间,之后逐渐缓和,最终结果与驯化结果相同。     

碱度胁迫对尼罗罗非鱼鳃离子细胞形态以及鳃、肾和肠中HCO3-转运因子的影响

采用扫描电镜观察了不同碱度(0、2、4 g/L Na HCO_3)胁迫对尼罗罗非鱼(Oreochromis niloticus)鳃离子细胞形态变化的影响,并采用免疫组化技术观察了鳃、肾、肠中4个HCO_3~-转运因子碳酸酐酶(CAⅡ、CAⅣ)、碳酸氢钠协同转运载体(SLC4A4)、Cl~-/HCO_3~-离子交换体(SLC26A6)的阳性反应变化。扫描电镜结果表明,鳃离子细胞分布在鳃小片基部。根据其表面开孔形状和尺寸,可分为Ⅰ型、Ⅱ型、Ⅲ型和Ⅳ型4种亚型,各亚型离子细胞的开孔尺寸随碱度胁迫强度增高呈正比增大,Ⅲ型离子细胞开孔尺寸变化最明显(P0.01);离子细胞总数目也随碱度升高而增加,Ⅲ型离子细胞数目上升最为显著(P0.01)。免疫组化结果表明,在淡水、碱水组中,CAⅡ、CAⅣ、SLC4A4、SLC26A6在鳃小片基部和肾中均有阳性反应,且随着碱度升高,阳性反应增强,但在肠道中未观察到阳性反应。本研究结果初步表明,尼罗罗非鱼可通过鳃离子细胞形态和数量调节适应碱度变化,鳃和肾为主要应答调节器官。     

纳米氧化锌对斑马鱼鳃组织的氧化损伤

正纳米颗粒是指粒径在1—100 nm,表面积较大,具有块状颗粒所没有的特有性质。由于纳米颗粒具有独特的理化性质,近年来合成和生产纳米颗粒的产量大大增加[1]。纳米氧化物颗粒在科研和工业产品的应用正在逐年增加,科学家预测依靠纳米技术创造的产品产生的价值,到2015年可能达到1万亿美元[2]。随着纳米颗粒的使用,它对人类健康和环境的风险也随之增加。因此了解纳米颗粒的不利影响,确定对生物的影响,确保纳米材料的安全使用是十分必要的[3]。     

Endothelin redistributes blood flow through the lamellae of rainbow trout gills

The lamellae of the fish gill are the primary sites for oxygen uptake from the water. Here, only two very thin layers of cells separate the blood from the water. Therefore, energetically costly ion-fluxes will also occur between blood and water, and it has been hypothesised that the blood flow within the lamellae can be regulated through vasoconstriction, but evidence for this has been lacking. Through direct observations of the lamellae of rainbow trout (Oncorhynchus mykiss) in vivo, using epi-illumination microscopy, we show here that an endothelium-derived vasoactive peptide, endothelin-1 (ET-1, 0.2 μg kg⁻¹ or 1.0 μg kg⁻¹), is able to completely constrict the vascular sheet in the lamellae, probably by inducing contraction of pillar cells. This coincided with a dose-dependent increase in ventral aortic blood pressure (rising from 6.6 kPa to 12.0 kPa in response to the high ET-1 dose). However, blood continued to flow through the marginal channel that circumvents each lamella. Thus, ET-1 caused an intralamellar blood shift from the lamellar sheet towards the marginal channels. Vasoconstriction in the lamellae is likely to provide the fish with a mechanism for matching its respiratory surface area with its respiratory needs, thereby minimising ion-fluxes. Accepted: 8 September 1998     

Differential responses in gills of euryhaline tilapia, Oreochromis mossambicus, to various hyperosmotic shocks

Euryhaline tilapia (Oreochromis mossambicus) survived in brackish water (BW; 20‰) but died in seawater (SW; 35‰) within 6 h when transferred directly from fresh water (FW). The purpose of this study was to clarify responses in gills of FW tilapia to various hyperosmotic shocks induced by BW or SW. In FW-acclimated tilapia, scanning electron micrographs of gills revealed three subtypes of MR cell apical surfaces: wavy-convex (subtype I), shallow-basin (subtype II), and deep-hole (subtype III). Density of apical surfaces of mitochondrion-rich (MR) cell in gills of the BW-transfer tilapia decreased significantly within 3 h post-transfer due to disappearance of subtype I cells, but increased from 48 h post-transfer because of increasing density of subtype III cells. SW-transfer individuals, however, showed decreased density of MR cell openings after 1 h post-transfer because subtype I MR cell disappeared. On the other hand, relative branchial Na⁺/K⁺-ATPase (NKA) α1-subunit mRNA levels, protein abundance, and NKA activity of the BW-transfer group increased significantly at 6, 12, and 12 h post-transfer, respectively. In the SW-transfer group, relative mRNA and protein abundance of gill NKA α1-subunit did not change while NKA activity declined before dying in 5 h. Upon SW transfer, dramatic increases (nearly 2-fold) of plasma osmolality, [Na⁺], and [Cl⁻] were found prior to death. For the BW-transfer group, plasma osmolality was eventually controlled by 96 h post-transfer by enhancement of NKA expression and subtype III MR cell. The success or failure of NKA activation from gene to functional protein as well as the development of specific SW subtype in gills were crucial for the survival of euryhaline tilapia to various hyperosmotic shocks.     

The Physiology and Evolution of Urea Transport in Fishes

This review summarizes what is currently known about urea transporters in fishes in the context of their physiology and evolution within the vertebrates. The existence of urea transporters has been investigated in red blood cells and hepatocytes of fish as well as in renal and branchial cells. Little is known about urea transport in red blood cells and hepatocytes, in fact, urea transporters are not believed to be present in the erythrocytes of elasmobranchs nor in teleost fish. What little physiological evidence there is for urea transport across fish hepatocytes is not supported by molecular evidence and could be explained by other transporters. In contrast, early findings on elasmobranch renal urea transporters were the impetus for research in other organisms. Urea transport in both the elasmobranch kidney and gill functions to retain urea within the animal against a massive concentration gradient with the environment. Information on branchial and renal urea transporters in teleost fish is recent in comparison but in teleosts urea transporters appear to function for excretion and not retention as in elasmobranchs. The presence of urea transporters in fish that produce a copious amount of urea, such as elasmobranchs and ureotelic teleosts, is reasonable. However, the existence of urea transporters in ammoniotelic fish is curious and could likely be due to their ability to manufacture urea early in life as a means to avoid ammonia toxicity. It is believed that the facilitated diffusion urea transporter (UT) gene family has undergone major evolutionary changes, likely in association with the role of urea transport in the evolution of terrestriality in the vertebrates.     

孕育对褶纹冠蚌滤食率的影响及鳃微结构变化

对比了褶纹冠蚌(Cristaria plicata)孕育蚌、未孕雌蚌和雄蚌的滤食率,并运用组织学、扫描电镜和透射电镜对实验蚌的鳃结构进行比较观察,以此探讨鳃结构变化对滤食功能的影响。滤食实验结果表明:孕育事件显著降低了雌蚌的滤食率,而未孕雌蚌与雄蚌的滤食率无显著差异。孕育雌蚌内外鳃均由两鳃小瓣愈合而成,每一鳃小瓣由成排的鳃丝组成,在中介区鳃丝之间通过丝间隔连接,在内部侧区则通过瓣间隔相连。雌蚌内鳃的鳃间隔为外鳃的2～3倍,而雄蚌的内外鳃无差异。孕育雌蚌外鳃在初级水管、瓣间隔等出现明显的变化,并出现了二级水管结构,而内鳃未发现显著变化。扫描电镜显示:在褶纹冠蚌鳃丝表面存在3种类型的纤毛(前纤毛、前侧纤毛和侧纤毛),其形态结构和分布各具特点,长径58～85μm椭圆形的鳃小孔成排相间分布于鳃丝之间,而3组实验蚌的内外鳃丝之间无明显差异。透射电镜观察发现:孕育雌蚌鳃丝表皮细胞表面形成突起,显著增加了表面微绒毛的数量,可能有利于雌蚌在孕育期间由于初级水管转化成育儿囊后对呼吸、滤食等功能的补偿,与其他蚌科物种报道类似。综合实验表明,孕育雌蚌外鳃结构的变化,尤其是初级水管的结构改变和二级水管、鳃丝表面褶皱的出现可能是影响孕育雌蚌滤食功能的主要原因。     

合浦珠母贝鳃的显微与超微结构

合浦珠母贝(Pinctada fucata)是典型的滤食性瓣鳃类动物,也是我国重要的海水珍珠养殖贝类。本研究用光学显微镜、扫描电镜和透射电镜观察了合浦珠母贝鳃的显微和超微结构。结果表明,合浦珠母贝鳃结构属于异丝鳃型,左右两侧各2个鳃瓣,每个鳃瓣由内鳃瓣和外鳃瓣组成。鳃瓣由主鳃丝和普通鳃丝构成,主鳃丝在鳃瓣中主要起支架作用,每2根主鳃丝之间的9～12根普通鳃丝由"簇内连接"(intrabunchial junction)相连成簇。普通鳃丝之间通过"丝间连接"(interfilament junction)相连,丝间连接的上皮细胞与普通鳃丝的扁平细胞结构一样,为鳃的呼吸上皮。丝间连接的存在扩大了鳃的表面积,这种结构有助于进行气体交换。主鳃丝和普通鳃丝表面有前纤毛和侧纤毛,与食物运送和气体交换有关。普通鳃丝表面的纤毛为典型的"9+2"型微管结构。     

台湾棘带吸虫囊蚴在鳙鳃丝、鳃耙上的分布

通过对鳙为期16个月的定期取样研究发现,1993年8月到1994年3月,台湾棘带吸虫的囊蚴主要分布在鳃耙上,1994年4月至11月,囊蚴则主要分布在鳃丝上;当鳙体长范围为30-90mm时,囊蚴主要分布在鳃耙上,当体长大于90mm时,囊蚴主要分布在鳃丝上,随着鳙的生长,鳃丝囊蚴所占的比例逐渐增大,从0增加到100%,而鳃耙囊蚴所占的比例逐渐减小,从100%下降到0;不同月份、不同体长组鳙其不同组鳃丝囊蚴分布规律相同,第2组鳃丝囊蚴分布较多,第3、第1、第4组鳃丝囊蚴分布依次减少：其不同组鳃耙囊蚴分布规律也相同,总是第1组鳃耙囊蚴分布较多,其余依次为第2、第3、第4组鳃耙;囊蚴在鳙鳃丝和鳃耙上的分布都呈聚集分布,且聚集的强度随囊蚴内种群密度的增高而增高。