Reduced hypoosmoregulatory ability and alteration in gill chloride cell distribution in mature chum salmon (Oncorhynchus keta) migrating upstream for spawning

Osmoregulatory ability of mature chum salmon (Oncorhynchus keta) during spawning migration was examined by following the changes in gill Na⁺, K⁺-ATPase activity and in the distribution and morphology of chloride cells. Mature chum salmon caught in Otsuchi Bay, northern Honshu Island, Japan, died within 5 d in seawater (SW) in association with a marked increase in plasma osmolality, whereas the fish transferred to fresh water (FW) maintained plasma osmolality efficiently. Gill Na⁺, K⁺-ATPase activity decreased in both SW-maintained and FW-transferred fish. Well-developed chloride cells, identified by immunocytochemical staining specific for Na⁺, K⁺-ATPase, were present mainly in the filament epithelium of immature fish caught in the ocean. In mature fish caught in the bay, however, additional chloride cells were also found in the lamellar epithelium. The number of filament chloride cells decreased markedly in the mature fish both in SW and in FW, whereas the number of lamellar chloride cells was maintained. These results suggest that the loss of hypoosmoregulatory ability in mature chum salmon may be attributable to the decrease in filament chloride cells and associated decrease in gill Na⁺, K⁺-ATPase activity, and also that appearance of lamellar chloride cells may be preparatory to the forthcoming upstream migration. Received: 14 April 1997 / Accepted: 5 May 1997     

Modulation of branchial ion transport protein expression by salinity in glass eels (<Emphasis Type="Italic">Anguilla anguilla</Emphasis> L.)

The Anguillid juvenile glass eel must deal with the osmoregulatory consequences of highly variable environmental salinities on its recruitment migration from coastal to fresh waters. Changes in ionoregulatory parameters and branchial ion transport protein [Na⁺/K⁺-ATPase, Na⁺:K⁺:2Cl⁻ cotransporter (NKCC), cystic fibrosis transmembrane regulator (CFTR) anion channel, V-type proton ATPase] expression (activities, protein and/or mRNA level expression and/or cellular localization) in response to acclimation to a broad range of ionic strengths [distilled water (DW) to hypersaline water (HSW; 150%) sea water (SW 32‰)] was studied. The estuarine glass eels were very euryhaline and successfully acclimated to acute changes in environmental ionic strength from 50% SW, with high mortality only observed in HSW (51%) and sublethal osmoregulatory indicators (whole body water content and sodium levels) disturbed at the extremes (DW and HSW). Central to a high salinity acclimation were elevated branchial Na⁺/K⁺-ATPase, NKCC and CFTR expression. At lower salinity, Na⁺/K⁺-ATPase expression was maintained and NKCC and CFTR expressions were reduced. Branchial chloride cells increased in size up to SW but decreased in HSW. During hypotonic disturbance (DW), no compensatory elevation in V-ATPase or Na⁺/K⁺-ATPase expression was observed.     

Temperature-dependent recruitment delay of the Japanese glass eel <Emphasis Type="Italic">Anguilla japonica</Emphasis> in East Asia

Japanese eels spawn mainly during June–August. The larvae (leptocephali) then drift for 3–5 months before metamorphosing into glass eels. The recruitment season generally starts in southern East Asia in November and in northern areas in April the following year, a lag of ~5 months. However, analysis of otolith daily growth rings revealed only a 1–2-month difference in the mean leptocephalus stage between southern and northern East Asian samples. Experiments and field observation indicate that glass eels may starve, lose body weight, and remain in early pigmentation stage for a few months in cold waters. The time lag in recruitment can be accounted for by a longer leptocephalus stage combined with a low temperature-driven delay to upstream migration in winter. The leptocephalus duration and oceanic currents determine the dispersal locations up to the glass eel phase, while temperatures determine the timing of upstream migration time at each location.     

Environmental levels of sediment-associated tri-n-butyltin chloride (TBTCl) and ionic regulation in flounders during seawater adaptation

The effects of exposure to sediment-associated tri-n-butyltin chloride (TBTCl) were examined in the euryhaline European flounder, Platichthys flesus (L.). The effects were quantified by measuring the changes in sodium efflux; Na⁺/K⁺-ATPase activity; and the numbers, areas, and distribution of chloride cells in the gills of freshwater-adapted fish, following a rapid transfer to seawater. Following the transfer to seawater, the Na⁺/K⁺-ATPase activity and the sodium efflux were significantly increased in the control group but remained unchanged in the TBTCl-exposed group. The normal morphological changes to the gill epithelium associated with seawater adaptation, which involve chloride cell distribution, took place in the control group but were significantly inhibited or delayed in the TBTCl group. The results presented in this study lead to the conclusion that environmental concentrations of tri-n-butyltin chloride in sediments are capable of significantly disrupting both the physiological and the morphological components of iono-regulatory functions of an estuarine flatfish.     

Branchial and intestinal osmoregulatory acclimation in the four-eyed sleeper, <Emphasis Type="Italic">Bostrychus sinensis</Emphasis> (Lacepède), exposed to seawater

Bostrychus sinensis is a facultative air breather that inhabits waters of a wide range of salinities. This study aimed to elucidate whether branchial and intestinal osmoregulatory acclimation occurred in B. sinensis transferred from 5‰ water through a progressive increase in salinities to seawater. Our results indicate that B. sinensis acted as a hyperosmotic regulator in 5‰ water, but exhibited hypoosmotic hypoionic regulation in seawater. During short- (1 day) and medium- (10 days) term acclimation to seawater, there were only minor perturbations in plasma osmolality and [Na⁺], which returned to control levels after 45 days of exposure to seawater. Branchial Na⁺/K⁺-ATPase activity was unaffected by 1, 10 or 45 days of exposure to seawater. However, prolonged (45 days) acclimation to seawater led to a significant increase in Na⁺/K⁺-ATPase α-subunit protein abundance. Taken together, these results indicate that there could be changes in the expression of Na⁺/K⁺-ATPase isoforms and/or post-translational modification of Na⁺/K⁺-ATPase in the gills of fish exposed to seawater. Immunofluorescence microscopy revealed that acclimation to seawater for 10 days only resulted in no change in branchial Na⁺/K⁺-ATPase protein expression, but there were increases in protein expression of cystic fibrosis transmembrane regulator (CFTR)-like chloride channel and Na⁺:K⁺:2Cl⁻ cotransporter (NKCC; probably NKCC1). Indeed, NKCC was undetectable in gills of fish kept in 5‰ water by Western blotting, but it became weakly detectable in fish exposed to seawater for 10 days and prominently expressed in fish exposed to seawater for 45 days. Therefore, our results indicate that branchial CFTR-like chloride channel and NKCC1 were the determining factors in the transition between hyperosmotic regulation and hypoosmotic hypoionic regulation in B. sinensis. Furthermore, the intestine of B. sinensis also served as an important osmoregulatory organ, since there were significant increases in both the activity and protein abundance of intestinal Na⁺/K⁺-ATPase in fish acclimated to seawater for 45 days. The effectiveness of branchial and intestinal osmoregulatory acclimation in B. sinensis during seawater acclimation led to only a minor increase in plasma osmolality, and thus resulted in relatively unchanged free amino acid contents in muscle and liver.     

A comparison of transport-related gill enzyme activities and tissue-specific free amino acid concentrations of Baltic Sea (brackish water) and freshwater threespine sticklebacks, Gasterosteus aculeatus, after salinity and temperature acclimation

The osmoregulatory abilities of one freshwater and two brackish water (Baltic Sea) populations of the euryhaline teleost fish Gasterosteus aculeatus were studied with respect to evolutionary physiology. Plasma osmolality, activities of Na⁺K⁺-ATPase, citrate synthase, creatine kinase in the gill and free amino acids in liver, axial muscle and pectoral fin muscle were measured. After transfer from 10 to 35 ppt at 15 °C, time-course changes of plasma osmolality and gill Na⁺K⁺-ATPase showed no significant fundamental differences between the freshwater and one of the Baltic Sea populations. In a multi-factorial experiment, each population was exposed to four different abiotic regimes. Both brackish water populations had high mortality in freshwater at 4 °C, which is discussed as a failure of osmotic regulation (reduced taurine concentrations). Freshwater specimens had higher levels of glycine in the axial and pectoral fin muscles compared to the brackish water populations. This is interpreted as a genetically based effect. In brackish (20 ppt) water of 15 °C, the freshwater population had high activities of Na⁺K⁺-ATPase, but low activities of creatine kinase, whereas both brackish water populations behaved in the opposite way. A fundamental difference between the freshwater and brackish water populations on the level of the osmoregulatory machinery was not observed. Received: 10 December 1998 / Accepted: 22 September 1999     

Microstructural growth in otoliths of conger eel (Conger myriaster) leptocephali during the metamorphic stage

Otolith growth during metamorphosis and some aspects of the early life history of conger eel (Conger myriaster) were determined as indicated from microstructure in otoliths of the leptocephali collected from Cheonsu Bay, Korea during May and June 1988. The leptocephali occurred from early May to late June in the study area. Larvae collected in early May were in the late leptocephalus stage, and the proportion of the metamorphosing leptocephali increased over time. Otoliths in the late leptocephalus stage showed a translucent zone only. Although the fish did not feed and the body length diminished during metamorphosis, the otolith continued to grow and, consequently, the opaque zone was formed outside the translucent zone. The inner translucent zone can be considered a leptocephalus zone, and the outer opaque zone a metamorphic zone. Assuming that the growth increments were deposited daily from hatching, the conger eel can be considered to have hatched between September and February. The number of increments in the inner hyaline zone ranged from 124 to 239, and was assumed to represent the number of days from hatching to the onset of metamorphosis. The duration of metamorphosis was estimated as 51 to 75 d based on the number of increments in the opaque zone at the end of the metamorphic stage.     

Effects of environmental hypercapnia on hemato-immunological parameters,carbonic anhydrase,and Na+, K+-ATPase enzyme activities in rainbow trout (Oncorhynchus mykiss) tissues

In this study, the effects of environmental hypercapnia on hemato-immunological parameters and the activities of respiratory enzymes such as carbonic anhydrase (CA) and Na⁺, K⁺-ATPase were investigated in rainbow trout (Oncorhynchus mykiss) tissues (gill, liver and kidney). Batches of 12 fish were exposed to 4.5 mg L^?1 (control) and 14 mg L^?1 CO₂. No mortalities occurred during the 14 days of the experimental period. Red blood cell (RBC), hemoglobin (Hb), and hematocrit (Ht) levels, and innate immune parameters such as nitro blue tetrazolium (NBT), lysozyme, and myeloperoxidase activities, and the melano-macrophage frequency were negatively affected by elevated CO₂ levels. Patterns of change in CA activity differed among the gill, liver, and kidney. Compared with the activities of CA in the control group, the CA enzyme was significantly stimulated at day 7 in the gill tissue, whereas it was stimulated at day 14 of the experiment in the liver tissue of fish exposed to 14 mg L^?1 CO₂ (P < 0.05). In contrast to the pattern of CA enzyme activities, the Na⁺, K⁺-ATPase enzymes were stimulated significantly in the liver after day 7 but inhibited in the kidney and gill (P < 0.05). These results suggest that a subchronic exposure to hypercapnia of rainbow trout tissues may lead to adaptive changes in the respiratory enzymes and negatively affects hemato-immunological parameters.     

Metabolic response under different salinity and temperature conditions for glass eel Anguilla japonica

Diadromous fish often enter freshwater directly from seawater via fish ladders or channels built in estuarine dams. The oxygen consumption rates (OCR) of glass eel, Anguilla japonica, were determined using an automatic intermittent flow respirometer under various salinity and temperature regimes to physiologically explain this direct movement. The endogenous rhythm of the OCR in wild glass eels, freshly collected from estuaries, was nearly synchronous with the tidal pattern at the estuarine collection site. When the salinity was changed from 20 psu (12°C) at a constant temperature to that of freshwater, the OCR of the glass eels decreased by 21.6±7.0% (mean ± SD) (P<0.05), showing a dampened rhythm for about 48 h. After this period of impediment, the glass eels resumed normal metabolic activity. Direct migration from seawater to freshwater under constant temperature would result in a severe physiological stress for these glass eels for about two days. When the glass eels were exposed to a cyclic change in water temperature of 2°C 26 h⁻¹, as they encounter in estuaries, and then were introduced to freshwater abruptly, the OCR rhythm corresponded to the cyclic changes in water temperature after exposure to freshwater. Under these conditions, the mean OCR of the glass eels had a small difference before and after exposure to freshwater. These data explained how glass eels can directly move from sea water into the freshwater without any apparent metabolic stress in the estuaries showing cyclic change in water temperature (Δt=2°C).     

Changes in otolith microstructure and microchemistry during larval development of the European conger eel (<Emphasis Type="Italic">Conger conger</Emphasis>)

Otolith microstructure and chemical composition (Sr:Ca ratios) of the European conger eel (Conger conger) were examined during the larval developmental stages by scanning electron microscopy and wavelength dispersive spectrometer. Back-calculated hatching dates from the otolith microstructure of the developing leptocephali indicate a protracted spawning season from December to June. The early age of our developing specimens captured south of the Azores Islands suggests that the conger eel has another spawning area closer to Azores than the Mediterranean. Otolith increment width, which was relatively constant and narrow in the developing leptocephalus stage, increased sharply at age 170-250 days. Sr:Ca ratios in the otolith, which increased during the developing leptocephalus stage, showed a rapid drop coinciding with the increase in increment width. These coincidental changes were regarded as the onset of metamorphosis for this species. A close linear relationship between the age at metamorphosis and otolith growth rate indicates that the faster-growing larvae metamorphose earlier, suggesting that somatic growth should play an important role in the timing of metamorphosis. As shown in earlier work, the existence of an otolith marginal zone with unclear rings during metamorphosis prevents an accurate estimate of the larval stage duration of this species.     

Sodium- and potassium-activated adenosine triphosphatase in the excretory organs of Sepia officinalis (Cephalopoda)

Sodium- and potassium-activated ATPase (Na⁺–K⁺-ATPase) has been demonstrated in excretory organs of Sepia officinalis, using a cytochemical procedure. In the renal appendages, both epithelia of the pancreatic appendages, the folded epithelium of the branchial heart appendage and the transport-active epithelium of the gill, the enzyme is localized exclusively in the basolateral cell membranes, i.e., the membranes of the basal labyrinth and the lateral plasma membranes. In addition, Na⁺–K⁺-ATPase is also located in the sarcolemma of the muscle fibres of the branchial heart. Distribution and localization of the enzyme is further substantiated by [³H]-ouabain autoradiography. The possible involvement of Na⁺–K⁺-ATPase in the excretion of ammonia and in ionic regulation in dibranchiate cephalopods is discussed.This study was supported by the Deutsche Forschungsgemeinschaft and is part of a doctoral dissertation     

Aspects of osmotic and lonic regulation in two Baltic teleosts: Effects of salinity on blood and urine composition

Hydromineral regulation was studied by examining the response to different environmental salinities in two Baltic brackish-water (BW) teleosts—a species of marine ancestry, Myoxocephalus scorpius (L.), and a glacial relict, M. quadricornis (L.). M. scorpius tolerated fresh water (FW) and M. quadricornis sea water (SW) for only about 24 h, but the survival time of M. scorpius in SW and M. quadricornis in FW was one to several weeks. M. scorpius seems able to balance plasma ionic concentrations in salinities down to about 2 to 3. Death of M. scorpius in FW was associated with partial haemolysis, increased volume of red blood cells (RBC), increased plasma K⁺ concentration, and decreased concentrations of Na⁺, Cl^- and Mg²⁺ in plasma and, to a lesser extent, in urine. Death of M. quadricornis in SW was associated with increased plasma osmolality, and Na⁺, Cl^- and Mg²⁺ concentrations, but the renal excretion of ions approached that generally found in marine teleosts. In most cases, RBC volume followed the changes in plasma osmolality or Na⁺ and Cl^- concentrations. Both species showed an ability to increase tubular Mg²⁺ secretion much over that needed in BW, and increased secretion was associated with high urine Cl^- concentration. M. quadricornis, but not M. scorpius, reabsorbed Na⁺ effectively in SW also. Differences between Oceanic and Baltic specimens of M. scorpius are discussed.     

溴氰菊酯对菲律宾蛤仔体内酶活性和组织损伤的初步探索

采用不同质量浓度的溴氰菊酯(0.0070 mg·L~(-1)、0.014 mg·L~(-1)、0.020 mg·L~(-1)、0.027 mg·L~(-1))对菲律宾蛤仔进行20 d半静置染毒,测定不同时间淋巴液中乙酰胆碱酯酶(ACh E)和钠离子-钾离子-三磷酸腺苷酶(Na~+-K~+-ATPase)活性、鳃和肝脏中谷胱甘肽转硫酶(GST)活性的变化,并观察染毒20 d后鳃丝组织和消化盲囊组织的损伤情况。酶活性分析结果显示,与对照组相比,低浓度组(0.0070 mg·L~(-1))试验期间酶活性均无显著差异(P0.05);中浓度组(0.014 mg·L~(-1)、0.020 mg·L~(-1))淋巴液中ACh E和Na~+-K~+-ATPase均呈先激活后抑制的变化规律(P0.05),鳃和肝脏中GST活性均呈上升趋势(P0.05);高浓度组(0.027mg·L~(-1))淋巴液中ACh E和Na~+-K~+-ATPase、肝脏中GST活性在试验期间持续下降(P0.01),而鳃中GST活性呈先抑制后升高的趋势(P0.05)。研究表明,低中浓度的溴氰菊酯对菲律宾蛤仔体内的酶活性表现为先诱导后抑制,具有明显的时间、剂量效应;高浓度的溴氰菊酯对菲律宾蛤仔体内酶活持续抑制,且染毒浓度越高,组织细胞变异越显著,表现为鳃丝上皮细胞纤毛层萎缩、纤毛脱落,消化盲囊上皮细胞膨胀,出现包涵体样结构。     

Gas–liquid chromatography for fenvalerate residue analysis: alterations in the ionic concentration and associated adinosine triphosphatases in fish,Cirrhinus mrigala (hamilton)

In the present study, an attempt has been made to quantify the fenvalerate accumulated in different tissues (gill, muscle and liver) and observe changes involved in the levels of sodium, potassium and calcium ions and Na⁺–K⁺, Mg²⁺ and Ca²⁺ adenosine triphosphatase (ATPase) activities in the freshwater fish, Cirrhinus mrigala on short-term and long-term exposure to the median lethal and sublethal concentration of fenvalerate. Residue analysis using gas–liquid chromatography (GLC) technique revealed that fenvalerate accumulated in highest quantity in gill followed by liver and muscle under median lethal concentration (6?µg?L^?1). Whereas in sublethal concentration (0.6?µg?L^?1), muscle accumulated highest quantity followed by gill and liver, which might be due to the fact that fenvalerate is highly lyphophilic. The ion concentration and ATPase activity were found effected in fish exposed to lethal and sublethal concentrations of fenvalerate. Concentration of Na⁺, K⁺ and Ca²⁺ ions decreased in gill, muscle and liver on being exposed to median lethal concentration to a significant level. Whereas the changes were not highly pronounced at sub lethal level indicating low concentration of fenvalerate and its non-toxic effect at chronic exposure. Na⁺–K⁺, Mg²⁺ and Ca²⁺ ATPases activity were also found decreased in correspondence to the ionic change under median lethal and sub lethal concentrations in target tissues. This might have lead to behavioural changes and create wide-spread disturbance in the normal physiology, ultimately causing the death of the fish. The results suggest that in biomonitoring programmes, ions and associated ATPases can be a good diagnostic tool for fenvalerate toxicity.     

Inhibition of Na+/K+-ATPase and of active ion-transport functions in the gills of the shore crab Carcinus maenas induced by cadmium

Inhibition of Na⁺/K⁺-ATPase from gill plasma membranes of the shore crab Carcinus maenas by cadmium was investigated and compared with inhibitory effects by known antagonists (ouabain and Ca²⁺). For comparative considerations the Cd²⁺-inhibition of the enzyme from dog kidney was also tested. Na⁺/K⁺-ATPase from dog kidney and from crab gill differed greatly in sensitivity against ouabain. The inhibition constant K _i of the dog enzyme amounted to 9.1 × 10⁻⁷ mol l⁻¹, i.e. more than 300-fold smaller than the K _i of 2.9 × 10⁻⁴ mol l⁻¹ determined for the crab enzyme. Ca²⁺ inhibited the activity of Na⁺/K⁺-ATPase from crab gill plasma membranes with a K _i of 4.3 × 10⁻⁴ mol l⁻¹. The Na⁺/K⁺-ATPase from crab gill was inhibited by Cd²⁺ with a K _i of 9.1 × 10⁻⁵ mol l⁻¹. Cd²⁺ inhibited the Na⁺/K⁺-ATPase from dog kidney with a K _i (6.4 × 10⁻⁵ mol l⁻¹) comparable to that observed in the crab gill enzyme. Under experimental conditions Cd²⁺-inhibition of Na⁺/K⁺-ATPase was irreversible. Repeated washing, centrifugation and homogenization of the plasma membranes (four times) with Cd²⁺-free buffer did not restore any activity lost in the presence of 1 × 10⁻³ mol l⁻¹ Cd²⁺. Since ouabain-insensitive (nonspecific) ATPases in the plasma membrane fraction of crab gills were inhibited by Cd²⁺ in the same way as Na⁺/K⁺-ATPase, the heavy metal is considered as an unspecific ATPase inhibitor. Comparing these results with literature data on Cd²⁺-binding to electrophoretically separated proteins suggests that Na⁺/K⁺-ATPase is a Cd²⁺-binding enzyme. The results obtained on Na⁺/K⁺-ATPase were reflected by Cd²⁺-inhibition of the branchial ion-transport functions depending on this enzyme. The transepithelial short-circuit current of isolated gill half lamellae, a direct measure of area-specific active ion uptake, and the transepithelial potential difference of isolated, perfused whole gills, also indicative of active ion uptake, were inhibited by the heavy metal in a time- and dose-dependent mode. Remarkably these inhibitions were also irreversible. These findings are ecologically and biomedically significant: even when the actual environmental or tissue concentrations measured are low, biological microstructures such as Na⁺/K⁺-ATPase may accumulate the heavy metal by tight binding over prolonged periods until the first inhibitory effects occur. Received: 25 June 1997 / Accepted: 25 August 1997     

Interaction of short-term testosterone treatment with osmotic acclimation in the gilthead sea bream <Emphasis Type="Italic">Sparus auratus</Emphasis>

To assess the interaction between testosterone (T) treatment and acclimation to different salinities, seawater-acclimated gilthead sea bream (Sparus auratus) were implanted with slow-release coconut oil implants alone (control) or containing T (5 μg/g body mass). After 5 days, eight fish of control and T-treated groups were sampled. The same day, eight fish of each group were transferred to low salinity water (LSW, 6 ppt, hypoosmotic test), seawater (SW, 38 ppt, control test) and high salinity water (HSW, 55 ppt, hyperosmotic test) and sampled 9 days later. Gill Na⁺, K⁺-ATPase activity increased in HSW-acclimated fish with respect to SW- and LSW-acclimated fish in both control and T-treated groups. Kidney Na⁺, K⁺-ATPase activity was also enhanced in HSW-acclimated fish, but only in T-treated group. From a metabolic point of view, most of the changes observed can be attributed to the action of salinity and T treatment alone, since few interactions between T treatment and osmotic acclimation to different salinities were observed. Those interactions included in treated fish: in the liver, decreased capacity in using glucose in fish acclimated to extreme salinities; in the gills, decreased capacity in using amino acids in HSW; in the kidneys increased capacity in using amino acids in extreme salinities; and in the brain, decreased glycogen and acetoacetate levels of fish in LSW.     

Early life history and recruitment of the tropical eel Anguilla bicolor pacifica, as revealed by otolith microstructure and microchemistry

Otolith microstructure and microchemistry of the tropical eel Anguilla bicolor pacifica Schmidt were examined in glass eels collected at the mouth of the Dumoga River, North Sulawesi Island, Indonesia. Ages of the glass eels examined (age at recruitment) ranged from 124 to 202 d (167 ± 19.3 d; mean ± SD), hatching being estimated as having occurred between November 1995 and March 1996. Otolith increment widths markedly increased from age 101 to 172 d (135 ± 18.2 d; mean ± SD), coincident with a drastic decrease in otolith Sr:Ca ratios, suggesting that metamorphosis began during that period. The duration of metamorphosis was estimated as 20 to 40 d, on the basis of otolith microstructural characteristics. The fluctuation patterns in otolith increment widths and Sr:Ca ratios were similar to those of the temperate Japanese eel A. japonica. Received: 20 May 1998 / Accepted: 7 October 1998     

三甲基氯化锡对斑马鱼(Danio rerio)生理生化特性的影响

为初步探讨三甲基氯化锡(trimethyltin chloride,TMT)对鱼类的毒性效应以及评价环境中TMT的安全性,采用静态鱼类急性毒性试验法测定了TMT对斑马鱼(Danio rerio)生理生化指标的影响;参考TMT的96 h-LC50值,设定3个浓度(0.39、0.78和1.17 mg·L-1)处理斑马鱼,测...     

Changes of plasma osmolality,chloride concentration and gill Na−K-ATPase activity in tilapia Oreochromis mossambicus during seawater acclimation

Changes of plasma osmolality, chloride concentration and gill Na–K-ATPase activity in tilapia Oreochromis mossambicus (obtained from Tainan Fish Culture Station of Taiwan Fisheries Research Institute, 1987) during seawater acclimation were examined. Three experiments were performed. (1) Freshwater (FW) to 30 salinity seawater (SW): plasma osmolality and chloride rose violently immediately post-transfer. At 6 h, gill Na–K-ATPase activity began to increase but most fish died from excessive plasma osmolality and Cl. (2) FW to 20 salinity SW: plasma osmolality and chloride increased immediately post-transfer, but more slowly than in (1), and began to decrease at 24 h. However it was not until 12 h post-transfer that gill Na–K-ATPase activity rose slowly. (3) FW to 20 salinity SW for 24 h, then to 30 salinity SW: after transfer to 30 salinity, plasma osmolality and chloride showed only a small increase initially then declined, while gill Na–K-ATPase activity started to rise rapidly within 3 h. The present results coincided with our previous morphological data concerning the ultrastructural responses of gill chloride cells. These are discussed to elucidate the osmoregulation mechanisms in tilapia during seawater acclimation.     

Target-site sensitivity in a specialized herbivore towards major toxic compounds of its host plant: the Na+K+-ATPase of the oleander hawk moth (Daphnis nerii) is highly susceptible to cardenolides

The caterpillars of the oleander hawk moth, Daphnis nerii (Linnaeus, 1758) (Lepidoptera: Sphingidae) feed primarily on oleander (Nerium oleander). This plant is rich in cardenolides, which specifically inhibit the Na⁺K⁺-ATPase. Since some insects feeding on cardenolide plants possess cardenolide-resistant Na⁺K⁺-ATPases, we tested whether D. nerii also possesses this strategy for circumventing cardenolide toxicity. To do so, we established a physiological assay, which allowed direct measurement of Na⁺K⁺-ATPase cardenolide sensitivity. Using Schistocerca gregaria, as a cardenolide-sensitive reference species, we showed that D. nerii Na⁺K⁺-ATPase was extremely sensitive to the cardenolide ouabain. Surprisingly, its sensitivity is even higher than that of the cardenolide-sensitive generalist, S. gregaria. The presence or absence of cardenolides in the diet of D. nerii did not influence the enzyme’s cardenolide sensitivity, indicating that target-site insensitivity is not inducible in this species. However, despite the sensitivity of their Na⁺K⁺-ATPase, caterpillars of D. nerii quickly recovered from an injection of an excessive amount of ouabain into their haemocoel. We conclude that D. nerii possesses adaptations, which enable it to feed on a cardenolide-rich diet other than that previously described in cardenolide specialized insects, and discuss other potential resistance mechanisms.