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31.
We focused on elucidating the toxic effects of water‐borne copper on the giant freshwater prawn Macrobrachium rosenbergii. After seven days of exposure to copper (Cu2+) at concentrations ranging from 0.01 mg L?1 to 0.5 mg L?1, three isozymes, malate dehydrogenase, alkaline phosphatase and esterase, were analyzed and compared using polyacrylamide electrophoresis (PAGE) and biochemical staining. The results indicated that the electrophoretic patterns of the isozymes showed a copper‐concentration‐related difference. Low doses of copper stimulated strong expression of the three isozymes. Electrophoretic patterns of malate dehydrogenase and alkaline phosphatase gradually became weaker or even lost as the level of copper increased. In contrast, esterase patterns exhibited an increased molecular heterogeneity at higher copper concentrations. A transmission electron microscope was used to study ultrastructure differences in the gills and hepatopancreas of M. rosenbergii, and the results showed significant structural damage at increased levels of copper compared with the control group. The basement membranes and mitochondira in the gills were seriously damaged, the cuticle electron density distribution was not homogeneous, and an infolded basement membrane, circularized nucleus, disintegrated nuclear membrane, and decreased mitochondria number and size were observed in the gills. Similarly, flowing out of karyoplasms, partly falling microvilli, decreased mitochondrion, partly disappeared mitochondrial cristae, and a thinned matrix were observed in the hepatopancreas. These findings indicate that exposure to elevated copper levels might damage the ultrastructure of the gills and hepatopancreas of M. rosenbergii and might further weaken their normal physical activities. Isozymes were quite sensitive to environmental stress and changes in isozyme elctrophoretic patterns might be effective biomarkers of environmental contamination.  相似文献   
32.
将VP110基因的部分序列克隆到pET-28a载体中构建pET28a-vp110b重组质粒并进行原核表达,获得重组表达的蛋白rVP110-B;用rVP110-B注射凡纳滨对虾Litopenaeus vannamei后,经WSSV感染,实验表明,该蛋白注射使凡纳滨对虾感染WSSV的半数死亡时间比对照组延长了20%。用表达纯化的该重组蛋白制备了兔抗rVP110-B多克隆抗体,该抗体用于凡纳滨对虾鳃细胞膜蛋白与rVP110-B的Far-western分析显示,凡纳滨对虾鳃细胞膜蛋白中除90 kDa左右的血蓝蛋白外,在41.7 kDa存在结合条带,经质谱分析表明这条鳃细胞膜蛋白是肌动蛋白。  相似文献   
33.
The myotomal fibres of the pectoral fins of white croaker (Micropogonias furnieri) have been studied using histochemical techniques and transmission electron microscopy (TEM). Succinic dehydrogenase (SDH) for mitochondria, periodic acid Schiff (PAS) for glycogen, Sudan Black for lipids and myosin-adenosintriphosphatase (mATPase) pre-incubated at alkaline and acid pHs were used to visualize the contraction velocity. Three zones were determined: superficial (SZ), medium (MZ) and deep (DZ). Staining for SDH, PAS and Sudan Black was positive only in the SZ. The level of alkaline mATPase was the highest in fibres from the DZ, intermediate in the MZ and low in the SZ; at an acid pH, the reverse was obtained. Fibres from the SZ were small with large quantities of subsarcolemmal mitochondria, scarce intermyofibrilar mitochondria and a well-developed sarcoplasmic reticulum; the myofibrils displayed a polygonal distribution along the entire length of the fibre. Fibres in the MZ were larger than those in the SZ, the myofibrils were densely packed, mitochondria prevailed under the sarcolemma and the sarcoplasmic reticulum was not abundant. Fibres from the DZ were the largest, with ribbon-shaped myofibrils and scarce mitochondria. The intercellular space was abundant and nervous endings were frequently observed.  相似文献   
34.
耳石微化学被广泛应用于鱼类洄游生态学研究,但耳石的摘取难免会造成鱼的死亡和损伤,故寻找可以快捷、非致死地替代耳石进行微化学分析的材料很有意义。为此,本研究比较了刀鲚(Coilia nasus)胸鳍条和耳石微化学的相似性特征。耳石的锶钙比值(Sr/Ca)和Sr含量图谱分型结果分别很好地对应了所研究的洄游型、淡水定居型和陆封型的典型个体。胸鳍条微化学结果显示,湖鲚和短颌鲚2类个体呈现出稳定的低Sr/Ca值?(1.38±0.52)~(2.04±0.74)和(1.40±0.32)~(1.81±0.66)?与刀鲚胸鳍条生活史第一阶段的特征相吻合?(2.06±0.64)~(2.26±0.29)?,这些均反映了淡水生境“指纹”的特征。刀鲚胸鳍条Sr含量图谱和耳石相似,自核心至边缘可分为明显的3个阶段,分别对应于面分布图上自核心至边缘的蓝色、绿色、蓝色之间微化学“指纹”的变化。由此可见,刀鲚胸鳍条可以作为耳石的非致死、低损伤的代替材料,用于刀鲚洄游习性、生境履历等研究。同时,由于胸鳍条具有取样和前处理的优势,在其他鱼类资源或标志放流评价研究时,可以考虑以胸鳍条代替耳石来开展研究。  相似文献   
35.
36.
Biochemical procedures developed to isolate plasma membranes from the branchial epithelium of rainbow trout (Oncorhynchus mykiss) yield membrane fractions that are specifically enriched in the plasma membrane marker enzyme Na+/K+-ATPase. As the bulk of the branchial Na+/K+-ATPase is assumed to be confined to the mitochondria-rich chloride cells, such membrane preparations must contain the essence of the enzymatic machinery of the chloride cells. Basal Na+ activity in branchial (chloride) cells is around 10 millimolar and, accordingly, we find a Km for Na+ of the Na+/K+-ATPase of 13 millimolar, indicating that the enzyme may be regulated by changes in cytosolic sodium. The Na+-gradient across the serosal plasma membrane created by this pump provides energy for 3Na+/Ca2+-exchange and bumetanide-sensitive Na+/K+/2Cl--cotransport. Here we further postulate the presence of a Na+/Cl--cotransporter, indicated by thiazide-sensitive, bumetanide-insensitive transport of Na+ and Cl-; this cotransporter activity awaits the characterization of its kinetics. The Na+/Ca2+-exchanger has kinetic characteristics compatible with a regulatory role of cytosolic Na+ in the activity of this carrier. Both Na+/Ca2+-exchange and Ca2+-ATPase activity may contribute to transport of Ca2+, the former having lower affinity for calcium but a higher capacity than the latter carrier. The Na+/K+/2Cl--cotransporter has kinetics that favor a regulatory role for plasma K+ in the activity of this carrier. Seawater adaptation leads to increased activity of cotransporter molecules in the plasma membrane fractions (the activity increases relative to that of the Na+/K+-ATPase) and this may reflect a function in Cl--extrusion performed by the chloride cells in a seawater environment. A function for the cotransporter in the gills of freshwater fish may be the regulation of cell volume.  相似文献   
37.
Goldfish, carp and trout gills were incubated with 3H-17-hydroxyprogesterone (17P). With goldfish gills, the metabolites were 17,20-dihydroxy-4-pregnen-3-one (17,20P; 82%), 17,20-dihydroxy-4-pregnen-3-one (17,20P; 8%), 11-ketotestosterone (KT) glucuronide (5.4%) and 17,20P glucuronide (0.2%). Sulfates were not detected. Carp gills converted 17P into 17,20P (11.2%), 17,20P (9.6%), KT (8.4%), glucuronides of 17,20P (1.3%) and 17,20P (1.6%) and sulfates of 17,20P (5.1%) and 17,20P (7.2%). 17,20P (38% free, 1.8% glucuronide and 21.1% sulfate) was the sole metabolite of 3H-17P in trout gill incubations. In the presence of high (10; µg ml-1) substrate concentration, cyprinid gills gave predominantly free 17,20P, while trout gills yielded only free 17,20P. Production of 17,20P, predominantly as its sulfate, from endogenous precursors was demonstrated in trout gills but was not stimulated by trout primary extract. Our results demonstrate for the first time the steroidogenic potential of teleost gills and suggest that they may play a role in secretion of pheromones in some species.  相似文献   
38.
Starry flounder (Platichthys stellatus) were cannulated and a bolus of 9 Ci14C-creatine in saline was injected into the caudal vein. The fish were sacrificed at intervals ranging from 1h to 36d after label injection. Creatine pool size (PCr+Cr) and creatinine (Crn) content in blood, muscle, gills and liver were analyzed and specific activities (SA) determined.Mean concentrations of PCr+Cr/Crn in PCA-extracts of muscle, gills, liver and blood of experimental fish (at rest) were 38.1/2.40, 4.1/0.25, 5.6/0.45 and 0.3/n.d. mol.g–1 respectively.Within 10 min, plasma SA had decreased by approximately 90%. In white muscle, the rate of14C–Cr appearance as well as label disappearance was slow compared to gills and liver. In fish examined 36d postinjection, mean SA in muscle had decreased to 23% of maximum SA which occurred 24h after injection.14C–Cr was incorporated into the liver tissue at a very high rate, SA being two orders of magnitude higher in liver than in white muscle. Over the first 6d, retention of label was observed in liver; after 36d only 3% of the original label was detected.Creatine pool size (PCr+Cr) in white muscle decreased with food deprivation. In flounder sacrificed after 36d, PCr+Cr was only 52% that of fed control fish, suggesting that creatine or precursors for its biosynthesis are supplied with the diet.  相似文献   
39.
Gas transfer and blood acid-base status in the blood-perfused trout head preparation (in vitro) were compared with intact trout (in vivo) fitted with oral membranes, dorsal aortic, ventral aortic, and opercular cannulae. Gas transfer rates were calculated from both arterial-venous blood differences and inspired-expired water differences using the Fick method. Oxygen uptake and carbon dioxide excretion were lower, while ammonia excretion was higher, in the blood-perfused head relative toin vivo rates. Gas transfer rates calculated from water were consistently greater than rates calculated from blood, the difference being greaterin vitro compared toin vivo. We conclude that the inadequacy of O2 and CO2 transfer in the blood-perfused head was not due to abnormal gill diffusive conductance, but was more likely related to the reduced magnitude of the blood-to-water O2 and CO2 diffusion gradients, low hematocrit, and decreased perfusion flow rate . Under the conditions of the present study, the blood-perfused trout head is not a suitable preparation for the study of oxygen transfer. We conclude this preparation is useful for evaluating branchial carbon dioxide or ammonia transfer only when comparable measurements or manipulations cannot be made on intact fish.  相似文献   
40.
Infectious gill diseases of marine salmonid fish present a significant challenge in salmon-farming regions. Infectious syndromes or disease conditions affecting marine-farmed salmonids include amoebic gill disease (AGD), proliferative gill inflammation (PGI) and tenacibaculosis. Pathogens involved include parasites, such as Neoparamoeba perurans, bacteria, such as Piscichlamydia salmonis and Tenacibaculum maritimum, and viruses, such as the Atlantic salmon paramyxovirus (ASPV). The present level of understanding of these is reviewed with regard to risk factors, potential impacting factors, methods of best practice to mitigate infectious gill disease, as well as knowledge gaps and avenues for future research.  相似文献   
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