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1.
FUMI  KATOH  TOYOJI  KANEKO 《Fisheries Science》2002,68(2):347-355
ABSTRACT: To examine the involvement of chloride cells in the uptake of Ca2+ in freshwater (FW) killifish, chloride cell morphology was compared in fish acclimated to different defined FW environments with Ca2+ concentrations of either 0.1 mM, 0.5 mM, or 2.5 mM. Numerous chloride cells were detected in whole-mount preparations of the gill filaments, which were stained with an antiserum specific for Na+, K+-ATPase. Chloride cells, located mostly on the afferent–vascular edge of the filaments, were larger at lower Ca2+ concentrations. Electron microscopic observations showed that in the 0.1 mM and 0.5 mM Ca2+ experimental groups, the apical membrane of chloride cells were flat or slightly projecting and equipped with numerous microvilli. In the 2.5 mM Ca2+ group, some chloride cells formed an apical pit, whereas other cells were similar to those observed in the 0.1 mM and 0.5 mM Ca2+ groups. Plasma osmolality decreased with decreasing ambient Ca2+ concentration, suggesting that environmental Ca2+ affects the permeability of the body surfaces. Gill Na+, K+-ATPase activity in the 0.1 mM and 0.5 mM Ca2+ groups were significantly higher than that in the 2.5 mM Ca2+ group, implying the involvement of the Na+–Ca2+ exchanger in Ca2+ uptake in the gills. These findings suggest that chloride cells function as the site for Ca2+ uptake in killifish acclimated to low Ca2+ environments.  相似文献   

2.
The effects of heavy metals (Hg2+, Cd2+, Co2+, Cr3+, Pb2+, Cu2+, Fe2+. Ni2+, Zn2+, Mn7+, Mn4+ and Mn2+) on the relative hatching percentage of brine shrimp, Artemia salina were assessed. Artemia salina cysts (San Francisco Bay Brand) hatched in seawater containing various heavy metal concentrations. After 48 hours, the number of hatched nauplii were counted. A negative linear relationship between hatching rate of Ariemia salina cysts and heavy metal concentration occurred, except copper which conformed to a cubic model. The relative toxicities of heavy metals on Artemia salina cysts were as follows (Cu2+, most toxic; Mn2+, least toxic): Cu2+ > Zn2+ > Ni2+ > Co2+ > Mn7+ > Cr3+ > Cd2+ > Fe2+ > Hg2+ > Mn4+ > Pb2+ > Mn2+.  相似文献   

3.
To determine the changes in blood chemistry associated with sequential transfer of summer flounder Paralichthys dentatus (320–480 g), 300 hatchery-reared fish were moved to three different environmental conditions during a 20-d period. Fish were transferred in progression from a recirculating seawater system (22 ppt, 22.5 C) to a flow-thru seawater system (31 ppt, 20.0 C), to three small coastal net pens (33 ppt, 15.5 C), and finally to a large open ocean net pen (33 ppt, 16.0 C). For this study, eight random fish were captured at each progressive step (environmental condition), anesthetized (MS222), and bled from the caudal vein (2 mL). Transferred flounder were bled every 12 h for 48 h to collect plasma cortisol and glucose samples. Fish were bled 24 h after transport and every 3 d thereafter for osmolarity, hematocrit, hemoglobin concentration, mean corpuscular hemoglobin content, glucose, cortisol, and the electrolytes Cl- Na+, K+ and Ca+. The most significant perturbations to blood chemistry (P < 0.05) occurred within 24 h of initial transfer from the recirculating to flow-thru seawater systems, suggesting an osmoregulatory rather than handling or transfer related stress. Osmolarity, electrolyte, and hematological parameters fluctuated and then recovered to stable levels by day 8 in the flow-thru seawater system. However, unlike the initial transfer, successive movement to the coastal and then the open ocean net pens produced transient increases in both plasma cortisol and glucose levels, suggesting a high level of stress associated with extended flounder handling and transfer.  相似文献   

4.
Growth and amino acid oxidation studies were conducted to estimate methionine requirement of juvenile Japanese flounder, Paralichthys olivaceus , by using the purified diets containing 500 g kg–1 crude protein from casein, gelatine and crystalline amino acids (CAA). Diets with six graded levels of methionine (5.3, 8.3, 11.3, 14.3, 17.3 and 20.3 g kg–1 diet) were fed to triplicate groups of the juvenile (initial weight 2.8 ± 0.05 g) twice a day for 40 days. To prevent leaching losses, CAA were precoated using carboxymethylcellulose (CMC), and further diets were bound by CMC and κ-carrageenan. Based on broken-line analysis of percentage weight gain and feed conversion efficiency, the methionine requirements of Japanese flounder in the presence of 0.6 g kg–1 of cystine were 14.9 and 14.4 g kg–1 dry diet, respectively. After the growth study was finished, a direct estimate of methionine requirement was made by examining the influence of dietary methionine level on 14C-methionine oxidation by determining radioactive carbon dioxide, protein and nonprotein fractions of the whole body. The dose–response curve between expired radioactive CO2 and dietary methionine levels showed that the optimum methionine level for the flounder was estimated to be within the range of 14.3–17.3 g kg–1 of diet in high agreement with values obtained from the growth study.  相似文献   

5.
Abstract. The sensitivity of the immunoperoxidase (IP) and fluorescent antibody (FA) techniques applied to frozen sections of organs of carp infected with spring viraemia virus (SVCV) was similar, both in respect of the intensity of the reaction and in the detection rate of the antigen. Seven days after a waterborne infection both methods detected the antigen in the kidneys and to a lesser extent in the liver and spleen. Quantification of virus gave a titre of 102.8 to 105.5 TCID50/g of kidney, whereas the agent could not be isolated from liver and spleen tissue. In contrast, at 241/2 days post–infection the viral antigen could be readily detected in liver and spleen but only occasionally in the kidneys using the IP and FA techniques. At this time, liver and spleen showed infectivity titres of 103.8 to 107.2 TCID50/g tissue, whereas the kidneys were found to be free of infective virus. It is concluded that using the IP and FA techniques SVCV can be detected most frequently in the kidneys from 7 to 14 days post–infection and in the liver and spleen from 14 to 24 days post-infection.  相似文献   

6.
Olive flounder artificially infected with lymphocystis disease virus (LCDV) were reared at 10, 20 and 30 °C for 60 days, to compare LCD-incidence. In the fish reared at 20 °C, lymphocystis cells appeared on the skin and fins at 35 days post-challenge, and the cumulative LCD-incidence was 80% at 60 days. High levels of LCDV, with a mean polymerase chain reaction (PCR) titre of 106 PCR-U mg−1 tissue, were detected in the fins and skin of LCD-affected fish at 20 °C, but were not detected in the spleen, kidney, brain and intestinal tissues of these fish. No LCD clinical signs were observed in the fish reared at 10 °C and 30 °C; however, a low level of LCDV (103 PCR-U mg−1 tissue) was detected in the fins and skin of these fish. By increasing the rearing temperature from 10 to 20 °C, lymphocystis clusters appeared on the skin and fins of the fish with no previous LCD clinical signs within 33 days after the temperature change. It was shown that permissive cells for LCDV infection exist in the epidermis of olive flounder. At low temperatures, small amounts of LCDV were able to persist over a period extended for a further 45 days in the fish epidermis, even though the fish showed no LCD clinical signs. The optimum growth temperature of LCDV is near 20 °C.  相似文献   

7.
ABSTRACT: The concentrations of Zn and sulfhydryl (SH) groups in the digestive tract tissue of common carp and some aquatic animals were studied. It was found that Zn and bound SH groups could be used as indicators for detecting the Zn-binding protein in the digestive tract tissue of common carp. The digestive tract tissue of the fish underwent subcellular fractionation, and it was found that the nuclei/cell debris fraction contained most of the DNA (85%), Na+/K+-ATPase (82%), organic phosphate (90%) and the Zn-binding protein (79%), but only part of the 5'-nucletidase and alkaline phosphatase (<23%). The nuclei/cell debris fraction of the digestive tract tissue of common carp was treated with either collagenase type I or type IV, and subfractionated by sucrose density centrifugation. It was found that treatment with collagenase type IV could release more than 50% of the Zn-binding protein, Na+/K+-ATPase and organic phosphate from collagen. Sections of digestive tract tissue of common carp were stained for Zn. It was observed that Zn can be found mainly on the edge of the epithelial layer, and everywhere in the 'membrane-like' portion of the submucosal and muscular layers. It is proposed that most of the Zn-binding protein in the digestive tract tissue of common carp is located on the basolateral plasma membranes of the epithelial cells and on the surrounding muscle cells that are attached to the collagen type IV of basal laminae.  相似文献   

8.
ABSTRACT: The filamentous fungi Rhizopus , like many fungal species, possesses physiologically active substances. Rhizopus extract (RU) is reported to be effective for various aspects of growth and reproduction in many vertebrates. The effects of RU administration on body growth and plasma levels of steroid hormones were investigated in lacustrine sockeye salmon Oncorhynchus nerka . One-year-old fish were fed daily with RU (20 mg/kg feed) from July 1999 to October 2000 for 15 months. Fish were sampled every month and plasma levels of testosterone (T), 11-ketotestosterone (11-KT), estradiol-17β (E2) and 17α,20β-dihydroxy-4-pregnen-3-one (DHP) were measured by time-resolved fluoroimmunoassay. Body growth of RU-fed fish of both sexes increased significantly in 1+ and 2+ October, and 2+ January–March and July. All RU-fed males and one female matured in 2+ October. RU-fed 1+ precociously mature males showed increased plasma levels of T, 11-KT and DHP in 1+ October. In 2+ males, RU significantly elevated plasma levels of T from May to June, 11-KT from June to July, and DHP in October. In sockeye salmon, administration of RU accelerated body growth of both sexes and sexual maturation in males, suggesting physiologically active substances present in RU enhance somatic growth and sexual maturation by sex-specific mechanisms.  相似文献   

9.
10.
The main objective of the present study was to evaluate whether dietary supplementation of urea might reduce the incidence of winter ulcer in sea water-farmed Atlantic salmon. Salmon destined to be S0 smolt were fed with a urea-supplemented diet (0 or 20 g kg−1 urea) over an 8-week period prior to sea water transfer and were then fed supplemented diet (0, 5, 10 or 20 g kg−1 urea) during the first and second winters in the sea. During the first winter positive relationships between dietary urea and plasma urea and between plasma urea and plasma osmolality were observed. Further, plasma osmolality displayed a negative relationship to mortality. Of the salmon that died during the first winter in the sea 90% had one or more skin ulcers. Both during the first and second winter there were fewer salmon with ulcer among fish fed with the diets supplemented with urea. Salmon fed with 20 g kg−1 urea tended to have higher percentage water in the muscle. Mortality and incidence of salmon with ulcer seemed to relate to plasma osmolality amongst fish fed on diets that differed in levels of urea supplementation, suggesting that an osmotic imbalance may contribute to the development of winter ulcer in farmed salmon. Salmon fed with 20 g kg−1 urea showed significantly greater body weight during the second winter in sea. Fish killed without prior starvation had significantly higher level of muscle urea in the 20 g kg−1 urea group compared with fish fed with the unsupplemented diet. However, a 13-day starvation period reduced urea content in the muscle to the level of the control. No effects of dietary urea supplementation on the sensory quality of market size Atlantic salmon were observed.  相似文献   

11.
Localization of Na+, K+-ATPase-rich cells in the gill and urinary system of Acipenser persicus fry was performed through immunofluorescence light microscopy using a mouse monoclonal antibody IgGα5 raised against the α-subunit of chicken Na+, K+-ATPase. Different types of epithelia were clearly identified in the gill epithelium: epithelia of branchial arch, interbranchial septum, filament and lamellar epithelium. The Na+, K+-ATPase-rich cells were found in the epithelia of branchial arch, interbranchial septum, filament, interlamellar region and also in the lamellae. Histologically, the urinary system is divided into head kidney, trunk kidney and short caudal kidney. The head kidney is composed of the pronephric tubules and the haemopoietic tissues, while the trunk kidney is composed of a large number of glomeruli and convoluted nephrons. Each nephron consisted of a large glomerulus and tubules (neck, proximal, distal and collecting tubules) which connected to ureters. Posteriorly, ureters extended and joined together to form a small urinary bladder. In the urinary system, no specific fluorescence staining was observed in the glomerulus, neck segment and proximal tubules. The distal tubule cells and collecting tubule cells showed a strong immunostaining of Na+, K+-ATPase. Epithelia of ureters and urinary bladder also showed several isolated immunofluorescent cells. Immunofluorescent cells were rich in Na+, K+-ATPase enzyme which is very important for osmoregulation.  相似文献   

12.
ABSTRACT: The present study was conducted to investigate the effects of dietary vitamin A on reproduction and egg quality in Japanese flounder Paralichthys olivaceus . Broodstock were fed experimental pellets containing two levels of vitamin A [11 × 103 IU/100 g (control diet; CD), 337 × 103 IU/100 g (experimental diet; ED)] for approximately 2 months before spawning and during the spawning period. Two groups of five females (average weight 1.4 kg) and 10 males (average weight 0.7 kg) were randomly allocated to two 30 m3 indoor tanks. Total egg production of the CD group was slightly higher than the ED group. Percentage of buoyant eggs and hatching rate of the ED group were significantly higher than the CD. In other egg quality parameters, such as percentage abnormal larvae and starvation tolerance of larvae, no notable difference was found between these two groups. At the end of the experiment, the skin color of broodstock in the ED group was darker than that of the CD group. Vitamin A content in eggs of the ED group was significantly higher than that of the CD group. However, the difference in vitamin A content in eggs between the ED and CD groups was much smaller than that in the liver of the females between the two groups. These results indicate that feeding broodstock a higher level of vitamin A increases the vitamin A content in eggs but does not affect egg quality in Japanese flounder because excess dietary vitamin A was stored mainly in the broodstocks' liver.  相似文献   

13.
Abstract Two groups of European catfish, Silurus glanis L., fingerlings were infected with channel catfish virus (CCV) by either intraperitoneal injection with 105 TCID50 of CCV, or bathing in water containing 105 TCID50 of CCV per 1·0 ml. The virus was isolated from spleen, intestine and brain of CCV-injected fish at day 1 and the titres ranged from 102·1 to 103·3 TCID50/g. However, the tissue distribution of CCV was irregular and no virus was isolated after day 3 post-exposure. In CCV-bathed fish, the virus was isolated only from the liver of one specimen at day 3 post-exposure. No clinical signs of CCV disease developed in any of the fish. Specimens in each regime from all sampling periods showed some minor histopathological changes, but there were no differences between treatments. Lesions included oedema and focal haemorrhage in the liver and the spleen was congested. Electron micrographs of tissue samples showed the presence of a few virus particles around the nuclei of kidney, spleen and intestinal cells, and in or around a myelinated nerve within the optic lobes of infected fish during the first 4 days of infection.  相似文献   

14.
ABSTRACT:   Amylase, with MW of 59 kDa, was purified from small abalone Haliotis sieboldii by ammonium sulfate fractionation, CM Sepharose Fast Flow and Sephacryl S-100 HR chromatographies. The optimal pH and temperature of purified amylase were 6.0 and 37°C, respectively. The purified enzyme was stable at pH 6.0–8.0 and low temperatures. It was activated by Ba2+, Mg2+, Ca2+, Co2+, Ni2+, Mn2+, K+, Ag+, Na+ and Li+, but completely or partially inhibited by Al3+, Cu2+, Cd2+, Hg2+ and Zn2+. EDTA could completely inhibit, while iodoacetamide, N-ethylmaleimide and urea partially inhibit the purified amylase. According to the digestion mode of various polysaccharides, the purified enzyme was considered to be an α-amylase.  相似文献   

15.
The aim of this study was to determine whether magnesium (Mg2+) in seawater is required for egg activation of the black tiger shrimp Penaeus monodon and whether manipulation of Mg2+ levels can be used to delay the process and thereby synchronize egg activation. Female P. monodon broodstock were allowed to spawn in artificial seawater containing Mg2+ at varying levels with respect to the normal (100%) level: 100%, 50%, 20% and 0%. Egg activation occurred normally at 100% Mg2+, incompletely at 50% and 20% Mg2+ levels and did not occur at all with 0% Mg2+. The fertilization rate with 100% Mg2+ was observed to be 83%, but fertilization failed to take place in all the other groups. The fertilization rate was restored from 0% to 76% following the 20% Mg2+ level treatment when Mg2+ levels returned to normal (100%) as soon as spawning was completed. This study suggests that the level of Mg2+ in seawater plays a vital role in P. monodon egg activation, and that commencement of this process could be delayed by manipulation of the Mg2+ level during and immediately after spawning.  相似文献   

16.
The influence of dietary cation–anion difference (CAD, Na+ + K+– Cl, mEq kg–1) on energy metabolism and nitrogen losses in juvenile African catfish Clarias gariepinus (Burchell) was examined in fish exposed to different dietary CAD levels (–146, 116, 497, 713 and 828 mEq kg–1 diet). The experiment was conducted in open circuit balance respiration chambers over a 3-week period. Five 24-h monitoring periods were carried out at 3-day intervals during the experimental period with O2 consumption, ammonia and nitrate + nitrite (NOx) and CO2 production being measured at 5-min intervals for each chamber. The negative dietary CAD (–146 mEq kg–1) resulted in the highest energy expenditures (83 kJ kg–0.8· d–1). With increasing dietary CAD levels, heat loss gradually decreased to minimum values of 56 kJ kg–0.8 day–1 at a dietary CAD level of 713 mEq kg–1. Consequently, metabolizable energy utilization efficiency (MEU, percentage of retained energy over metabolizable energy) quadratically ( P  < 0.05) increased and reached a maximum at a dietary CAD of 713 mEq kg–1.  相似文献   

17.
Seasonal variations in osmoregulatory and metabolic parameters were assessed in juvenile gilthead sea bream ( Sparus auratus ) cultured in earthen ponds under a natural photoperiod and temperature. Specimens were sampled, and the plasma, gill, kidney and liver were collected during winter 2005 and 2006 (January), spring 2005 (April), summer 2005 (July) and autumn 2005 (October). Plasma osmoregulatory parameters showed higher values in summer, while metabolic parameters presented different patterns of variations. Gill Na+,K+-ATPase activity decreased significantly in winter, while gill metabolite levels showed different patterns of variations among seasons. The enzymatic activities tested did not present a clear pattern of variation [(glutamate dehydrogenase (EC 1.4.1.2) (GDH) and hexokinase (EC 2.7.1.11) (HK)] or significant differences along seasons [glucose 6-phosphate dehydrogenase (EC 1.1.1.49)]. Kidney Na+,K+-ATPase activity decreased during summer and autumn. Different patterns of variation were observed in kidney metabolite levels while all the enzymatic activities assessed [lactate dehydrogenase-oxidase (EC 1.1.1.27) (LDH-O), HK and GDH] presented the highest values during summer. In the liver, metabolite levels and enzymatic activities did not show significant variations or present clear patterns of variation along different seasons. These results indicated seasonal variations in the osmoregulatory and metabolic parameters of different organs (blood, gill, kidney and liver) in earthen pond-cultured gilthead sea bream ( S. auratus ), which could be mainly attributed to seasonal changes in temperature.  相似文献   

18.
Abstract The role of A-layer (A), protease (P) and haemolysin (H) as virulence factors of Aeromonas salmonicida, the aetiological agent of fish furunculosis, was a investigated using three strains of the bacterium. Strain MT004 lacked the A-Iayer (A) and produced extracellular caseinase and gelatinase (P+) and haemolysin (T-lysin; H+). Strain MT028 was A, P and H, and strain MT048 was A+, P+ and H. The pathology and LD50 produced in rainbow trout by cells or extracellular products (ECP) of each strain were determined. The ECP was produced by two different methods where the protease and haemolytic activities differed in relative levels, or when the protease of MT004 ECP was inhibited by the serine protease inhibitor PMSF. The results indicate that the presence of A-layer is not essential, at least for a moderate degree of virulence; that in vitro production of extracellular proteases is not a requisite of virulent strains; that presence of protease and haemolysin in ECP can be correlated with the development of certain lesions and a rapid time to death but cannot be correlated with the lethal toxicity of the ECP. The authors conclude that an as-yet unrecognized component of ECP is responsible for killing fish.  相似文献   

19.
ABSTRACT:    This study incorporated the 43 kDa Zn-binding membrane protein isolated from common carp into liposome. The specificity and strength of the binding of 65Zn to the 43 kDa protein-liposomes, and the binding of the 65Zn-labeled 43 kDa protein-liposomes to laminin were studied. It was found that 65Zn was bound to the external side of the 43 kDa protein-liposomes. Specific binding of 65Zn to the protein-liposomes was detected. The binding parameter of Zn to the protein was found to be: maximum binding site (Nmax), 76.7 pmole/µg protein (approx. 3 mole of Zn2+/mole); and equilibrium dissociation constant (Kd), 0.19 µM. Of the cations introduced (Ca2+, Cd2+,Co2+, Cr2+, Cu2+, Fe2+, Hg2+, Mg2+, Mn2+, Ni2+, Pb2+), only Co2+ competed significantly with Zn. The protein-liposomes were also found to bind specifically to laminin with a Nmax of 1.1 pmole/µg laminin, and Kd of 4.79 µM. No significant protein-liposome binding occurred to other extracellular matrix proteins (fibronectin, fibrinogen or vitronectin). Furthermore, the binding was specifically inhibited by the Arg-Gly-Asp (RGD) peptide or GRGDSPG, while two other analogs (GRGESPG and GRADSPG) were without effect.  相似文献   

20.
迟恒  绳秀珍  唐小千  战文斌 《水产学报》2009,33(6):1011-1017
应用抗牙鲆免疫球蛋白单克隆抗体(2D8、1H1)对牙鲆外周血系统、肾、脾、肝、胰、肠道组织中抗体阳性细胞进行了定位观察,并对其组织学特征进行了描述。两株单抗均能在外周血滴片和组织切片中成功地检测到抗体阳性细胞。外周血系统中的抗体阳性细胞主要为淋巴细胞,没有发现抗体阳性的巨噬细胞;牙鲆头肾中没有肾单位,肾小管、肾小球等主要存在于后肾中,脾脏和肾脏都含有巨噬细胞、粒细胞、淋巴细胞等免疫相关细胞,抗体阳性细胞存在方式也极为相似,成簇或单独分布于黑色素巨噬细胞中心和血管周围;牙鲆的胰组织镶嵌在肝上,形成肝胰脏,也参与免疫应答,抗体阳性细胞单个存在,分布于肝组织中,胰组织中没有发现抗体阳性细胞;肠道抗体阳性细胞主要存在于固有层中,有成簇存在现象,在上皮层也可见到单个存在的抗体阳性细胞。  相似文献   

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