Acclimation of brackish water pearl spot (Etroplus suratensis) to various salinities: relative changes in abundance of branchial Na+/K+-ATPase and Na+/K+/2Cl− co-transporter in relation to osmoregulatory parameters

The present study was conducted to elucidate the osmoregulatory ability of the fish pearl spot (Etroplus suratensis) to know the scope of this species for aquaculture under various salinities. Juvenile pearl spot were divided into three groups and acclimated to freshwater (FW), brackish water (BW) or seawater (SW) for 15 days. The fish exhibited effective salinity tolerance under osmotic challenges. Although the plasma osmolality and Na⁺, K⁺ and Cl^? levels increased with the increasing salinities, the parameters remained within the physiological range. The muscle water contents were constant among FW-, BW- and SW-acclimated fish. Two Na+/K+-ATPase α-isoforms (NKA α) were expressed in gills during acclimation in FW, BW and SW. Abundance of one isoform was up-regulated in response to seawater acclimation, suggesting its role in ion secretion similar to NKA α1b, while expression of another isoform was simultaneously up-regulated in response to both FW and SW acclimation, suggesting the presence of isoforms switching phenomenon during acclimation to different salinities. Nevertheless, NKA enzyme activities in the gills of the SW and FW individuals were higher (p < 0.05) than in BW counterparts. Immunohistochemistry revealed that Na⁺/K⁺-ATPase immunoreactive (NKA-IR) cells were mainly distributed in the interlamellar region of the gill filaments in FW groups and in the apical portion of the filaments in BW and SW groups. The number of NKA-IR cells in the gills of the FW-acclimated fish was almost similar to that of SW individuals, which exceeded that of the BW individuals. The NKA-IR cells of BW and SW were bigger in size than their FW counterparts. Besides, the relative abundance of branchial Na⁺/K⁺/2Cl^? co-transporter showed stronger evidence in favor of involvement of this protein in hypo-osmoregulation, requiring ion secretion by the chloride cells. To the best of our knowledge, this is the first study reporting the wide salinity tolerance of E. suratensis involving differential activation of ion transporters and thereby suggesting its potential as candidate for fish farming under different external salinities.     

Effect of exposure to low salinity water on plasma ion regulation and survival rates in artificially wounded devil stinger <Emphasis Type="Italic">Inimicus japonicus</Emphasis>

We investigated the effect of exposure to low salinity water on plasma ion regulation and survival rates in artificially wounded devil stinger Inimicus japonicus. All fishes survived in 33% seawater (SW), while survival rate in 100% SW was 5.1% at 24 h. In 100% SW, plasma Na⁺, K⁺, Mg²⁺, and Ca²⁺ concentrations significantly increased to 238?±?49.9, 9.6?±?2.4, 15.1?±?3.5 and 5.0?±?0.7 mmol/l at 6 h, respectively; the gill Na⁺/K⁺–ATPase (NKA) activity was almost stable, although only one fish survived to 24 h. In 33% SW, plasma Na⁺ and K⁺ concentrations remained at the same level, and plasma Mg²⁺ and Ca²⁺ concentrations gradually increased to 16.2?±?0.7 and 4.5?±?0.2 mmol/l until 24 h, respectively. The NKA activity significantly increased to 5.1?±?1.1 µmol ADP/mg protein per h at 6 h. A positive correlation was observed between the wound surface area against body weight and the plasma ion concentrations, although no difference was observed in the restoration rate of the wounded area between 100 and 33% SW. These results indicate that exposure of wounded fish to low salinity water improves survivability by favoring plasma ion regulation without influencing the restoration rate.     

急性盐度胁迫对摄食不同镁水平饲料鲈血清渗透压和离子水平以及鳃丝ATP酶活力的影响

为研究饲喂不同镁水平饲料后,鲈血清渗透压、离子水平和鳃丝ATP酶活力对急性盐度胁迫的反应,实验用镁水平为0.413 g/kg(D1,对照组)、1.042 g/kg(D2)、1.577 g/kg(D3)、1.991 g/kg(D4)的4种实验饲料,分别投喂初始平均体质量为(30±0.5)g的鲈。实验鱼在淡水循环养殖系统中饲养50 d后立即转入海水循环养殖系统,并在转入前(0 h)和转入后1、3、6、12、24 h分别采集鲈鳃丝和血液样本。结果显示:盐度胁迫前饲料镁水平对鲈血清渗透压、Na+、K+、Cl-、Mg2+含量和鳃丝中Na+/K+-ATP(NKA)与Ca2+/Mg2+-ATP(CMA)酶的活力有显著影响(P<0.05),其中血清渗透压、Mg2+含量和NKA酶活力随饲料镁水平的增加而升高。D4组的血清渗透压、Mg2+含量、NKA和CMA酶活力显著高于其他组(P<0.05),表明该组鲈处于高强度的渗透压调节的生理状态。急性盐度胁迫中,血清Na+和Cl-含量随胁迫时间的延长呈逐渐上升趋势,血清渗透压、K+、Ca2+、Mg2+和鳃丝ATP酶活力在0～24 h内呈波动性变化。D1组的NKA和CMA酶活力在胁迫1 h时显著低于其他组(P<0.05),表明长期摄食低镁水平的饲料会降低鲈鳃丝NKA和CMA酶对环境盐度刺激的敏感度。研究表明,淡水养殖鲈的饲料中镁水平应低于1.991 g/kg,以减少维持鳃丝NKA与CMA酶高活力而导致的能量损失,而在环境由低盐向高盐的快速转变过程中,较高的饲料镁水平可通过快速提高的鳃丝NKA与CMA酶活力而有利于鲈快速适应高盐环境。     

Physiological effects of salinity on Delta Smelt, <Emphasis Type="Italic">Hypomesus transpacificus</Emphasis>

Abiotic factors like salinity are relevant to survival of pelagic fishes of the San Francisco Bay Estuary. We tested the effects of 4 parts per thousand (ppt) salinity increases on Delta Smelt (DS) in a laboratory experiment simulating salinity increases that might occur around the low-salinity zone (LSZ) (<6 ppt). Adult DS, fed 2 % body mass per day, starting at 0.5 ppt [freshwater (FW)], were exposed to weekly step-increases of 4 ppt to a maximum of 10 ppt saltwater (SW) over 19 days, and compared to FW controls. DS (n = 12/treatment per sampling) were sampled at 24, 72, and 96 h (1, 3, and 4 days) post-salinity increase for analyses of hematocrit, plasma osmolality, muscle water content, gill chloride cell (CC) Na⁺/K⁺-ATPase (NKA) and apoptosis after being weighed and measured (n = 3 tanks per treatment). No apparent increase in length or weight occurred nor did a difference in survival. Following step-increases in SW, hematocrit increased over time. Other fish responses generally showed a pattern; specifically plasma osmolality became elevated at 1 day and diminished over 4 days in SW. Percent muscle water content (%) did not show significant changes. CCs showed increased NKA, cell size and apoptosis over time in SW, indicating that CCs turnover in DS. The cell renewal process takes days, at least over 19 days. In summary, DS are affected by salinities of the LSZ and ≤10 ppt, though they employ physiological strategies to acclimate.     

Exploration of the mechanisms of protein quality control and osmoregulation in gills of Chromis viridis in response to reduced salinity

Fish gills are the vital multifunctional organ in direct contact with external environment. Therefore, activation of the cytoprotective mechanisms to maintain branchial cell viability is important for fish upon stresses. Salinity is one of the major factors strongly affecting cellular and organismal functions. Reduction of ambient salinity may occur in coral reef and leads to osmotic stress for reef-associated stenohaline fish. However, the physiological responses to salinity stress in reef-associated fish were not examined substantially. With this regard, the physiological parameters and the responses of protein quality control (PQC) and osmoregulatory mechanisms in gills of seawater (SW; 33–35 ‰)- and brackish water (BW; 20 ‰)-acclimated blue-green damselfish (Chromis viridis) were explored. The results showed that the examined physiological parameters were maintained within certain physiological ranges in C. viridis acclimated to different salinities. In PQC mechanism, expression of heat-shock protein (HSP) 90, 70, and 60 elevated in response to BW acclimation while the levels of ubiquitin-conjugated proteins were similar between the two groups. Thus, it was presumed that upregulation of HSPs was sufficient to prevent the accumulation of aggregated proteins for maintaining the protein quality and viability of gill cells when C. viridis were acclimated to BW. Moreover, gill Na⁺/K⁺-ATPase expression and protein amounts of basolaterally located Na⁺/K⁺/2Cl^? cotransporter were higher in SW fish than in BW fish. Taken together, this study showed that the cytoprotective and osmoregulatory mechanisms of blue-green damselfish were functionally activated and modulated to withstand the challenge of reduction in salinity for maintaining physiological homeostasis.     

iTRAQ proteomic analysis of salinity acclimation proteins in the gill of tropical marbled eel (<Emphasis Type="Italic">Anguilla marmorata</Emphasis>)

Osmoregulation plays an important role in the migration process of catadromous fish. The osmoregulatory mechanisms of tropical marbled eel (Anguilla marmorata), a typical catadromous fish, did not gain sufficient attention, especially at the molecular level. In order to enrich the protein database of A. marmorata, a proteomic analysis has been carried out by iTRAQ technique. Among 1937 identified proteins in gill of marbled eel, the expression of 1560 proteins (80 %) was quantified. Compared with the protein expression level in the gill of marbled eel in freshwater (salinity of 0 ‰), 336 proteins were up-regulated and 67 proteins were down-regulated in seawater (salinity of 25 ‰); 33 proteins were up-regulated and 32 proteins were down-regulated in brackish water (salinity of 10 ‰). These up-regulated proteins including Na⁺/K⁺-ATPase, V-type proton ATPase, sodium–potassium–chloride co-transporter and heat shock protein 90 were enriched in many KEGG-annotated pathways, which are related to different functions of the gill. The up-regulated oxidative phosphorylation and seleno-compound metabolism pathways involve the synthesis and consumption of ATP, which represents extra energy consumption. Another identified pathway is the ribosome pathway in which a large number of up-regulated proteins are involved. It is also more notable that tight junction and cardiac muscle contraction pathways may have correlation with ion transport in gill cells. This is the first report describing the proteome of A. marmorata for acclimating to the change of salinity. These results provide a functional database for migratory fish and point out some possible new interactions on osmoregulation in A. marmorata.     

Photoperiod and temperature effects on growth and maturation of pre- and post-smolt Atlantic salmon

The present study was undertaken to examine the long-term effect of continuous light and constant temperature and their interaction on growth, feed intake, gill Na⁺, K⁺-ATPase (NKA) activity and early sexual maturation in Atlantic salmon pre- and post-smolts. The fish (mean initial weight = 15.9 g ± 0.4 SE) were reared on two photoperiods (continuous light, LL and simulated natural photoperiod, LDN, 60°25′N) and on two constant temperatures (average 8.3 and 12.7 °C) from June to July the following year. This resulted in four experimental groups abbreviated as LL8, LL12, LDN8 and LDN12. Growth in freshwater was highest in the LL12 group and final weight of this group was 70–330 % higher than in the other experimental groups, and our findings further demonstrate that the growth-enhancing effect of continuous light alone corresponds to a 4.5° increase in temperature. Overall, the highest feed intake was registered in the LL12 group, whereas no differences in feed intake or growth were observed between the LL8 and LDN12 groups, and the lowest feed intake and growth in the LDN8 group. Both temperature groups on LL developed peak levels in gill NKA activity in October–November, 4–5 months prior to the natural season for the parr–smolt transformation. Fish at 12 °C showed peak levels in NKA activity 4–6 weeks before the fish on 8 °C. The proportion of mature males was higher at 12.7 °C (66 %) compared to 8.3 °C (11 %). Highest maturation was seen at LL12 (82 %). For the salmon industry, this means that long-term rearing at LL and 12.7 °C will improve growth but also leads to higher maturation proportion. By rearing fish at LL8, it is, however, possible to achieve high growth and low maturation simultaneously.     

Immunolocalization of chloride transporters to gill epithelia of euryhaline teleosts with opposite salinity-induced Na<Superscript>+</Superscript>/K<Superscript>+</Superscript>-ATPase responses

Opposite patterns of branchial Na⁺/K⁺-ATPase (NKA) responses were found in euryhaline milkfish (Chanos chanos) and pufferfish (Tetraodon nigroviridis) upon salinity challenge. Because the electrochemical gradient established by NKA is thought to be the driving force for transcellular Cl⁻ transport in fish gills, the aim of this study was to explore whether the differential patterns of NKA responses found in milkfish and pufferfish would lead to distinct distribution of Cl⁻ transporters in their gill epithelial cells indicating different Cl⁻ transport mechanisms. In this study, immunolocalization of various Cl⁻ transport proteins, including Na⁺/K⁺/2Cl⁻ cotransporter (NKCC), cystic fibrosis transmembrane conductance regulator (CFTR), anion exchanger 1 (AE1), and chloride channel 3 (ClC-3), were double stained with NKA, the basolateral marker of branchial mitochondrion-rich cells (MRCs), to reveal the localization of these transporter proteins in gill MRC of FW- or SW-acclimated milkfish and pufferfish. Confocal microscopic observations showed that the localization of these transport proteins in the gill MRCs of the two studied species were similar. However, the number of gill NKA-immunoreactive (IR) cells in milkfish and pufferfish exhibited to vary with environmental salinities. An increase in the number of NKA-IR cells should lead to the elevation of NKA activity in FW milkfish and SW pufferfish. Taken together, the opposite branchial NKA responses observed in milkfish and pufferfish upon salinity challenge could be attributed to alterations in the number of NKA-IR cells. Furthermore, the localization of these Cl⁻ transporters in gill MRCs of the two studied species was identical. It depicted the two studied euryhaline species possess the similar Cl⁻ transport mechanisms in gills.     

LHRHa‐induced ovulation of the endangered‐Caspian brown trout (Salmo trutta caspius) and its effect on egg quality and two sex steroids: testosterone and 17α‐hydroxyprogestrone

To induce synchronized ovulation, migrating wild Caspian brown trout (Salmo trutta caspius) females were treated with two interperitoneal injections of Des‐Gly¹⁰, d ‐Ala⁶ LHRH (LHRHa), given 3 days apart. Two injections of 100 μg kg^?1 body weight of this hormone effectively induced ovulation. Within 27 days from the second injection, all fish injected with 100 μg kg^?1 LHRHa had ovulated compared with 54.5% of the controls. The mean time to ovulation was reduced significantly (P<0.05) from 31.67±4.84 days in control fish and 28.83±7.31 days in sham‐treated fish to 16.36±1.61 days in fish injected with 100 μg kg^?1 LHRHa. The fertilization rate in 50 and 100 μg kg^?1 LHRHa‐injected fish was significantly lower than that in the control fish (P<0.05). In fish injected with 50 and 100 μg kg^?1 LHRHa, significant (P<0.05) changes in testosterone (T) and 17α‐hydroxyprogestrone (OHP) levels were observed. After the second LHRHa injection, the fish injected with 100 μg kg^?1 showed the highest serum levels of testosterone and OHP. These results demonstrate that the use of LHRHa can effectively reduce the mean time to ovulation and induce synchronized ovulation in Caspian brown trout.     

不同盐度和驯养时间中华鲟子二代幼鱼鳃的显微结构变化

采用光学显微镜和组织切片方法,研究了不同盐度和驯养时间中华鲟幼鱼鳃及鳃中泌氯细胞的结构变化特征。试验用鱼为1+龄中华鲟子二代,平均全长50.7 cm,平均体重约441.9 g。试验前在养殖池中暂养1周。试验设2个盐度梯度(10、15)组和1个对照组(淡水)。盐度驯养为将中华鲟子二代从暂养池转入盐度10海水中驯养,分别取驯养1、3、6、9、12、24、48、72、96、120、144、168、192、216、240和264 h的幼鱼各1尾用于试验;将剩下的鱼再转入盐度15海水中驯养,分别取驯养24、48、72 h的幼鱼各1尾用于试验。结果显示,在淡水组中,中华鲟子二代幼鱼鳃丝发达,细胞饱满,鳃小片宽且长。鳃丝上皮的泌氯细胞数量较少,通常靠近鳃小片基部,呈近椭圆状。与淡水组相比,盐度10组的幼鱼鳃丝主干部明显变窄,鳃小片基部区域及鳃小片上泌氯细胞的数量略有增加,胞体变大,且随驯养时间的延长,出现了明显的细胞膜间隙。幼鱼在盐度15组中鳃丝主干明显变窄变短,鳃小片也显著缩短,鳃泌氯细胞数量显著增加,主要分布在鳃小片基部,有明显的分泌腔。中华鲟子二代幼鱼能够快速适应外界水体盐度的变化,这与鳃及鳃上皮中调节渗透压功能细胞的数量和结构变化相适应。     

Fluctuating salinity effect on Sphoeroides annulatus (Jenyns 1842) physiological responses

Euryhaline fish, such as the Bullseye puffer Sphoeroides annulatus (Jenyns 1842), experience sudden salinity changes in their natural environment, which is more common than the exception, so they must adapt to survive and cope with extreme salt conditions. Therefore, Bullseye puffer juveniles were exposed to short‐term stress (39 hr) by fluctuating salinity conditions (41, 35, 29, 23, 17, 11, 5, 11, 17, 23, 29, 35, 41 psu) with a 3‐hr interval between each point at 26 ± 1ºC in a respirometer chamber and acclimation reservoirs. Responses to oxygen consumption rate (OCR: 23–35 mg O₂ h^–1 kg^–1), ammonium excretion rate (AER: 1–1.85 mg NH4⁺ h^?1 kg^?1), oxygen‐nitrogen atomic ratio (O:N 17–30), osmoregulatory pattern (blood osmotic pressure from 342.4 to 332.8 mmol/kg) and changes in expression levels of Na⁺/K⁺‐ATPase in the gills (higher values at higher salinities) were measured. Although some signs of stress were detected below the iso‐osmotic point (11.4 psu), the puffer fish is a strong euryhaline fish that survives under these conditions. Nonetheless, it could recover when salinity returned to the initial acclimation point because Sphoeroides annulatus is able to live in a wide range of environments with wide natural salinity fluctuations; thus, a common practice in aquaculture has been to expose fish to low salinity for several reasons discussed in this study. This capacity reveals its high plasticity to saline adaptation from 41 to 5 psu an up from 5 to 41 psu, all in less than 2 days.     

Salinity and temperature tolerance of brown-marbled grouper Epinephelus fuscoguttatus

Grouper have to face varied environmental stressors as a result of drastic changes to water conditions during the storm season. We aimed to test the response of brown-marbled grouper to drastic and gradual changes in temperature and salinity to understand the grouper’s basic stress response. The results can improve the culture of grouper. Brown-marbled grouper, Epinephelus fuscoguttatus (6.2 ± 0.8 g) were examined for temperature and salinity tolerances at nine different environmental regimes (10, 20, and 33 ‰ combined with 20, 26 and 32 °C), in which the fish were subjected to both gradual and sudden changes in temperature and salinity. The critical thermal maximum (50 % CT_MAX) and the upper incipient lethal temperature (UILT) were in the ranges of 35.9–38.3 and 32.7–36.5 °C, respectively. The critical thermal minimum (50 % CT_MIN) and the lower incipient lethal temperature (LILT) were in the ranges of 9.8–12.2 and 14.9–22.3 °C, respectively. The critical salinity maximum (50 % CS_MAX) and the upper incipient lethal salinity (UILS) were in the ranges of 67.0–75.5 and 54.2–64.8 ‰, respectively. Fish at temperature of 20 °C and a salinity of 33 ‰ tolerated temperatures as low as 10 °C when the temperature was gradually decreased. Fish acclimated at salinities of 10–33 ‰ and a temperature of 32 °C tolerated salinities of as high as 75–79 ‰. All fish survived from accumulating salinity after acute transfer to 20, 10, 5, and 3 ‰. But all fish died while transferred to 0 ‰. Relationships among the UILT, LILT, 50 % CT_MAX, 50 % CT_MIN, UILS, 50 % CS_MAX, salinity, and temperature were examined. The grouper’s temperature and salinity tolerance elevated by increasing acclimation temperature and salinity. On the contrary, the grouper’s temperature and salinity tolerance degraded by decreasing acclimation temperature and salinity. The tolerance of temperature and salinity on grouper in gradual changes were higher than in drastic changes.     

Response of ATPases in the osmoregulatory tissues of freshwater fish Oreochromis niloticus exposed to copper in increased salinity

An increase in salinity of freshwater can affect the physiology and metal uptake in fish. In the present study, Nile tilapia Oreochromis niloticus were exposed to copper (1.0 mg/l) in increased salinities (2, 4, and 8 ppt) for 0, 1, 3, 7, and 14 days. Following the exposures, the activities of Na⁺/K⁺-ATPase, Mg²⁺-ATPase, and Ca²⁺-ATPase were measured in the gill, kidney, and intestine to evaluate the changes in osmoregulation of fish. Results showed that increases in salinity and Cu exposure of fish significantly altered the ATPase activities depending on the tissue type, salinity increase, and exposure durations. Salinity-alone exposures increased Na⁺/K⁺-ATPase activity and decreased Ca²⁺-ATPase activity. Na⁺/K⁺-ATPase activity decreased following Cu exposure in 2 and 4 ppt salinities, though the activity increased in 8 ppt salinity. Ca²⁺-ATPase activity decreased in the gill and intestine in all salinities, while the activity mostly increased in the kidney. However, there were great variations in Mg²⁺-ATPase activity following exposure to salinity alone and salinity+Cu combination. Cu accumulated in the gill and intestine following 14 days exposure and accumulation was negatively correlated with salinity increase. Data indicated that ATPases were highly sensitive to increases in salinity and Cu and might be a useful biomarker in ecotoxicological studies. However, data from salinity increased freshwaters should carefully be handled to see a clear picture on the effects of metals, as salinity affects both metal speciation and fish osmoregulation.     

Effect of dietary protein levels and feeding rates on the growth and health status of juvenile genetically improved farmed tilapia (<Emphasis Type="Italic">Oreochromis niloticus</Emphasis>)

A 2 × 3 factorial study was conducted to investigate the effects of dietary protein levels (DPLs) and feeding rates (FRs) on the growth and health status of juvenile genetically improved farmed tilapia (GIFT), Oreochromis niloticus. Triplicate tanks of fish (initial weight 15.87 ± 0.11 g) were fed diets containing 25 or 35% protein at rates of 3, 5, or 7% body weight per day (BW day^?1) for 8 weeks. At the end of the feeding trial, the results showed that fish growth (final mean weight 34.61–81.07 g) and condition factor (3.39–4.45 g cm^?3) increased with the DPLs and FRs. Feed efficiency (FE, 0.48–0.88) increased as DPLs increased but decreased as FRs increased; the opposite trend was observed for feed cost (FC, 3.24–5.82 CHN Yuan kg^?1) and hepatosomatic index (0.98–2.33%). Apparent protein retention efficiency (APRE, 23.92–38.78%) was reduced by high FR. A 35% protein diet resulted in higher (P < 0.05) FE and APRE, and lower (P < 0.05) FC at 5% BW day^?1 than those at 7% BW day^?1. As FRs increased, lipid contents of the hepatopancreas, viscera, muscle, and eviscerated body increased, while moisture contents of hepatopancreas and viscera decreased. All serum biochemical parameters, including glutamic-oxalacetic transaminase and glutamic-pyruvic transaminase activity and levels of creatinine, glucose, high-density lipoprotein cholesterol, low-density lipoprotein cholesterol, total cholesterol, triglyceride, and total protein were unaffected by DPL or FR (P > 0.05), except urea nitrogen levels, which were affected by DPLs (P < 0.05). Moreover, the size of hepatocytes and the area ratio of hepatocyte vacuoles were enlarged (P < 0.05), whereas the area ratios of the nucleus and cytoplasm were reduced (P < 0.05) with increasing FRs. These results suggested that the optimal feeding strategy for juvenile GIFT is 35% protein diet at 5% BW day^?1.     

Effect of dietary chitosan on growth performance,hematological parameters,intestinal histology and stress resistance of Caspian kutum (<Emphasis Type="Italic">Rutilus frisii kutum</Emphasis> Kamenskii, 1901) fingerlings

This study was conducted to investigate the effects of dietary chitosan on growth performance, hematological parameters, intestinal histology, stress resistance and body composition in the Caspian kutum (Rutilus frisii kutum, Kamenskii, 1901) fingerlings. Fish (1.7 ± 0.15 g) were fed diets containing chitosan at different levels (0, 0.25, 0.5, 1 and 2 g kg^?1 diet) for a period of 60 days. Results showed that the feed conversion ratio significantly decreased in fish fed diet containing 1 g kg^?1 of chitosan compared to the other groups (P < 0.05), but there were no significant differences between treatments in terms of specific growth rate and condition factor (P > 0.05). Leukocyte increased in fish fed diet containing 2 g kg^?1 of chitosan compared to the other groups (P < 0.05). Lymphocytes, eosinophils and neutrophils did not significantly change among dietary treatments (P > 0.05). Also, the chitosan supplementation did not affect the whole-fish body composition (P > 0.05). Light microscopy demonstrated that the intestinal villus length increased in fish fed diet containing 1 g kg^?1 of chitosan compared to control group (P < 0.05). While 11 and 13 ‰ salinity and 30 °C thermal stress had no effect, 1 g kg^?1 of chitosan (P < 0.05) showed highest survival rate (70 %) in 34 °C thermal stress. The results showed that chitosan in the diet of the Caspian kutum fingerlings could improve feed conversion ratio, the nonspecific defense mechanisms and resistance to some of the environmental stresses.     

Osmoregulation in juvenile Chinese sturgeon (<Emphasis Type="Italic">Acipenser sinensis</Emphasis> Gray) during brackish water adaptation

The osmoregulation capabilities of 7-month-old juvenile Chinese sturgeon (Acipenser sinensis Gray) (128.8 ± 15 g) transferred directly from fresh water (0‰, 46 mOsmol kg⁻¹) to brackish water (10‰, 273 mOsmol kg⁻¹) were studied over a 20-day period. Changes in serum osmolarity, chloride (Cl⁻), sodium (Na⁺), potassium (K⁺) and calcium (Ca²⁺) ion concentrations, as well as gill and spiral valve Na⁺,K⁺-ATPase activities were measured at 3, 12, 24, 72, 216 and 480 h after transfer to BW. The serum osmolarity and ion concentrations (Na⁺, Cl⁻ and Ca²⁺) increased immediately after the transference to BW, reaching maximum at 24 h and returned to a new steady state at 216 h, while the FW control group maintained basal levels which showed lower (P < 0.05) than the BW group. Gill Na⁺,K⁺-ATPase activity of BW group exhibited an abrupt decrease in the first 3 h after transfer, but began to increase at 3 h, reaching a peak value at 24 h, and returned to a new steady state at 216 h. The differences between gill Na⁺,K⁺-ATPase activity of BW and FW fish were significant (P < 0.05) after 12 h. In contrast, Na⁺,K⁺-ATPase activity of the spiral valve showed transient increase after transference from FW to BW, and then decreased rapidly at 3 h, reaching the lowest at 24 h after transference. At 216 h after exposure to BW, Na⁺,K⁺-ATPase activities of the spiral valve increased slowly to the levels of FW control. The results of our study indicate the existence of hyposmoregulatory adaptive mechanisms in 7-month-old juvenile Chinese sturgeon which enable this fish to acclimate itself successfully to brackish water.     

Growth performance,osmoregulatory and metabolic modifications in red porgy fry,Pagrus pagrus,under different environmental salinities and stocking densities

The effects of two different environmental salinities [brackish water (BW), 12‰; sea water (SW), 39‰] and initial stock densities [low (LD), 1.0 g L^?1; high (HD), 2.0 g L^?1] on growth, osmoregulation, stress and energy metabolism of the fry Pagrus pagrus were investigated over a period of 45 days. Pagrus pagrus (n=80, 5.51 ± 0.25 g mean initial body weight) were randomly divided in eight groups. Growth, weight gain and specific growth rate increased in BW‐acclimated fish compared with SW‐acclimated fish. No differences were observed between the two stock densities tested at either environmental salinity. Plasma osmolality was lowest in BW‐acclimated specimens, but the stock density had no effect on this parameter. Branchial Na⁺,K⁺‐ATPase activity was positively correlated with environmental salinity, but unaltered at the renal level. Plasmatic parameters were enhanced by salinity and stocking conditions. At the hepatic level, triglyceride values were enhanced in BW‐acclimated fish maintained at LD. Muscle metabolites (glycogen, glucose and lactate) increased in BW‐ compared with SW‐acclimated fish; stock density had no influence. Our data suggest that changes in metabolic parameters could be correlated with the higher growth rates observed in P. pagrus acclimated to BW, while no significant effects due to the stocking density used were observed.     

Dietary resveratrol improves immunity but reduces reproduction of broodstock medaka <Emphasis Type="Italic">Oryzias latipes</Emphasis> (Temminck &amp; Schlegel)

Here, we investigated the effect of dietary resveratrol (20, 40, and 80 µg/g BW/day) on cell-mediated immunity (activity of spleen phagocytes and proliferative response of lymphocytes) and reproductive parameters (egg and sperm quality, i.e. fecundity—total number of eggs produced by individual fish, fertility, embryo survival, and hatching rate) in medaka. Fish fed feed with resveratrol at 40 and 80 µg/g BW/day had significantly higher metabolic activity and intracellular phagocyte killing activity than control. The proliferative lymphocyte activity of the fish from R80 group was greater by more than 20 % in comparison with the control group (P < 0.05). The percentage of macrophages (MO) and their mean fluorescence intensities (MFI) in R40 and R80 groups were significantly higher compared to C and R20 groups (P < 0.05). The differences in MO and MFI values ranged from 52.5 % (±1.5; R0 group) to 65.8 % (±1.6; R80 group) and from 23.2 (±1.4; R0 group) to 38.2 (±2.4; R80 group), respectively. Moreover, resveratrol at 80 µg/g BW/day decreased liver COX activity, i.e. 5.4 in R80 group and 7.9 in R0 group (P < 0.05). The motility parameters of the sperm obtained from the males fed feed supplemented with resveratrol at 80 µg/g BW/day exhibited the highest values except the linearity, which was lower as compared to the control (P < 0.05). The results indicate that diet supplemented with resveratrol at a dosage of 40 µg/g BW/day improves phagocyte killing ability and lymphocyte proliferation in broodstock and accelerates offspring hatch. Also, the results suggest that COX activity influences sperm and oocyte quality in fish; the presence of a COX inhibitor in the dose of 40 µg/g BW/day decreased the embryo survival.     

Spawning induction in Kutum Rutilus frisii kutum (Kamenskii, 1901) using carp pituitary extract or GnRH analogue combined with metoclopramide

Kutum Rutilus frisii kutum (Kamenskii, 1901), Cyprinidae is an endemic fish of the Caspian Sea. Iranian Fisheries Organization (Shilat) produce up to 200 million fry (1–2 g body weight (b.w.)) to restock the Caspian Sea population annually. Some of these fry are produced by spawning induction in broodfish by carp pituitary extract (CPE). The objective of this study was to assay the effectiveness of the gonadotropin releasing hormone analogue (d ‐Ala⁶, Pro⁹‐Net GnRH) alone or in combination with metoclopramide (MET), a dopamine antagonist, on the percentage of ovulated females, latency period, ovulation index and fertilization success. The following hormone treatments were tested: single injection of 2 mg kg^?1 b.w. of CPE as a positive control, GnRHa alone 20 and 40 μg kg^?1 b.w. and combination of GnRHa and MET as follows: 5 μg+2.5 mg, 10 μg+ 5 mg and 20 μg+10 mg kg^?1 b.w. Negative control group was injected with 0.7% saline. The percentage of ovulated females, ovulation index and fertilization success were 90%, 71.3±1.24%, 68.4±2.3%, respectively, in the group treated with GnRHa+MET at a dose of 20 μg+10 mg kg^?1 b.w. and were significantly higher than those in the positive control (60%, 64.5±0.23%, 69.1±4.5%) (P<0.05). However, the latency period in this group was longer than that in the positive control (P<0.05). Only 20% and 40% fish ovulated in groups that received 20 or 40 μg kg^?1 b.w. GnRHa. No fish ovulated in the negative control.