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.     

Effects of sodium selenite on some biochemical and hematological parameters of rainbow trout (Oncorhynchus mykiss Walbaum, 1792) exposed to Pb2+ and Cu2+

This study was carried out to understand the preventive effect of selenium (Se⁴⁺) on heavy metal stress induced by lead and copper in rainbow trout (Oncorhynchus mykiss). Variation in glutathione peroxidase (Se-GSH-Px) and superoxide dismutase (SOD) activity, and in malondialdehyde (MDA) levels in liver, spleen, heart, and brain tissues of rainbow trout after 72 h of exposure to Pb²⁺ and Cu²⁺ were investigated in the presence and absence of Se⁴⁺. In the presence of Se⁴⁺, Se-GSH-Px activity and SOD activity were found to be higher and MDA levels were lower compared with in its absence. Hematological parameters were also determined and it has been observed that total leukocyte count (WBC), mean cell volume (MCV), and mean cell hemoglobin (MCH) were increased and erythrocyte number (RBC), hemoglobin (Hb), and hematocrit value (Hct; P < 0.05) were decreased in fish exposed to heavy metals in the absence of selenium. Selenium presence recovered hematological parameters to normal levels. In the light of our findings, it could be stated that Pb²⁺ and Cu²⁺ lead to dramatic changes in biochemical and hematological parameters and selenium caused these parameters to converge to control levels when it was administered concurrently with these heavy metals.     

Responses of antioxidant enzymes,lipid peroxidation,and Na<Superscript>+</Superscript>/K<Superscript>+</Superscript>-ATPase in liver of the fish <Emphasis Type="Italic">Goodea atripinnis</Emphasis> exposed to Lake Yuriria water

Lake Yuriria, located in the heavily populated and polluted Mexican Central Plateau, receives domestic sewage, industrial effluents, and municipal wastewaters that are still directly discharged without treatment into the tributaries and the lake. Pollutants in water and sediments include heavy metals, aromatic hydrocarbons, and organochlorine pesticides. Activities of the antioxidant enzymes superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx), as well as Na⁺/K⁺-adenosine triphosphatase (Na⁺/K⁺-ATPase) activity, and lipid peroxidation (LPO) were evaluated in the livers of the fish Goodea atripinnis after 96 h of exposure to water collected in March and June 2005 from three sites: Y (limnetic zone), L (Lerma tributary), and C (la Cinta tributary). Physical and chemical parameters of the lake water were also analyzed. Increases in CAT activity and LPO levels at all three sites were detected compared with control fish (P < 0.05), while GPx and SOD activities decreased significantly (P < 0.05). Na⁺/K⁺-ATPase activities were similar to the control in fish exposed to limnetic water from both March and June but were higher than control at the two tributary sites in March (P < 0.05); fish exposed to water from the Lerma tributary in June exhibited lower Na⁺/K⁺-ATPase than the control (P < 0.05). During March, CAT and Na⁺/K⁺-ATPase activities were increasing more than in June in Y and L, respectively, while in June, SOD and GPx were depleted more than March in L and Y and L, respectively. Despite the antioxidant defenses of the fish liver, exposure to all water samples from Lake Yuriria exerted alterations in hepatic LPO levels, antioxidant enzymes, and Na⁺/K⁺-ATPase activities that could substantially impair the mechanisms of fish defenses against oxidative stress.     

Mitochondrion-rich cells distribution,Na<Superscript>+</Superscript>/K<Superscript>+</Superscript>-ATPase activity and gill morphometry of the Amazonian freshwater stingrays (Chondrichthyes: Potamotrygonidae)

Detailed measurements of gill area and constituent variables (total filament number, total filament length and mean filament length), and immunolocalization of the α-subunit of Na⁺/K⁺-ATPase and Na⁺/K⁺-ATPase activity were performed on both hemibranchs of all five arches of freshwater potamotrygonid stingrays (Paratrygon aiereba and Potamotrygon sp.). Both species exhibit similar mass-specific gill area, 89.8 ± 6.6 and 91.5 ± 4.3 mm² g⁻¹ for P. aiereba and Potamotrygon sp., respectively. The density of Na⁺/K⁺-ATPase-rich MRCs and Na⁺/K⁺-ATPase activity was higher in the 4th gill arch in both species. The Na⁺/K⁺-ATPase activity was positively correlated to the Na⁺/K⁺-ATPase-rich (Na⁺/K⁺-ATPase rich) mitochondrion-rich cell (MRC) distribution among the gill arches of P. aiereba but not in Potamotrygon sp. The levels Na⁺/K⁺-ATPase activity were not correlated to the gill surface area among the arches for both rays’ species. Considering that the Na⁺/K⁺-ATPase-rich MRC is the main site for active ion transport in the gill epithelia and Na⁺/K⁺-ATPase activity plays a crucial role in osmoionoregulatory function, we suggesting that 4th gill arch is more relevant for osmoregulation and ion balance in these potamotrygonids.     

Mechanism of Cd<Superscript>2+</Superscript> on DNA cleavage and Ca<Superscript>2+</Superscript> on DNA repair in liver of silver crucian carp

The subject of acute injury, apoptosis and canceration of animals induced by heavy metal ions has been one of the hotspots studied worldwide. However, the exact molecular mechanism of Cd-induced carcinogenicity remains largely unclear, and how to relieve the toxicity in vivo has rarely been reported. For this paper, we have investigated the mechanism of Cd²⁺ on DNA cleavage and Ca²⁺ on DNA repair in the liver of silver crucian carp (Carassius auratus gibelio) by agarose gel electrophoresis methods and by estimating biochemical indexes. Our results show that Cd²⁺ induces the classical laddering degradation of DNA in vivo and that DNA cleavage is repaired after injection with Ca²⁺ under various Cd²⁺ concentrations. DNA cleavage caused by Cd²⁺ is due to the activation of deoxyribonuclease (DNase) and the accumulation of reactive oxygen species (ROS). Furthermore, Cd²⁺ destroys the antioxidant system, which diminishes the activities of superoxide dismutase (SOD), catalase (CAT) and peroxidase (POD), causing an increase of the lipid peroxidation (LPO) level, respectively. However, after the liver is injected with Ca²⁺ under various Cd²⁺ concentrations, the DNase activity, the ROS generating rate and the LPO level are obviously reduced, the activities of SOD, CAT, and POD are greatly increased. At the same time, ROS production and removal recoves its balance. The results show that Ca²⁺ can relieve the toxicity of Cd²⁺ in silver crucian carp.     

Osmoregulation in juvenile Siberian sturgeon (Acipenser baerii)

The osmoregulation capabilities of juvenile Siberian sturgeon exposed to three experimental osmolalities (22, 250 and 387 mOsmol kg⁻¹) were studied over a 45-day period. Growth performance, haematological parameters, ion concentrations, gill and spiral valve Na⁺-K⁺-ATPase activities, as well as gill and spiral valve histology, were measured. At the end of the period, the plasma osmolality of fish kept in 250 and 387 mOsmol kg⁻¹ was higher than that of fish kept in 22 mOsmol kg⁻¹. Similar trends were observed in electrolyte concentrations. Spiral valve and gill Na⁺-K⁺-ATPase activity varied with exposure time and environmental salinity. Shortly after being transferred to hyperosmotic media, spiral valve Na⁺-K⁺-ATPase activity fell, while gill Na⁺-K⁺-ATPase activity remained constant. At the end of the experiment, gill Na⁺-K⁺-ATPase activities in fish kept in isosmotic and hyperosmotic media had increased in comparison to those of the control fish. Moreover, spiral valve Na⁺-K⁺-ATPase activities recovered and were similar to those recorded in fish kept in hyposmotic environments. Although some of the morpho-physiological mechanisms were operational in juvenile Siberian sturgeon in their adaptation to hyperosmotic media, fish cannot be considered hyperosmotic regulators as they were unable to maintain their plasma osmolality and electrolyte equilibrium in salinities higher than 250 mOsmol kg⁻¹. This suggests that the culture of juvenile Siberian sturgeon in brackish environments is unlikely to be successful. However, our data indicate that in natural environments, juvenile Siberian sturgeon in migratory populations (Ob and Lena Rivers) would be able to migrate successfully into estuarine brackish grounds with a salinity of up to 9%. This revised version was published online in July 2006 with corrections to the Cover Date.     

盐度胁迫对三疣梭子蟹鳃Na+/K+-ATPase酶活的影响

通过钼蓝法测定三疣梭子蟹在3组实验盐度的胁迫过程中第2对和第6对鳃Na⁺/K⁺-ATPase酶活的变化,比较了3组实验盐度胁迫1 d时,鳃Na⁺/K⁺-ATPase的酶活大小。结果表明,在盐度胁迫初期,3组实验盐度下第2对和第6对鳃Na⁺/K⁺-ATPase的酶活下降;之后,各组实验盐度下第2对和第6对鳃Na⁺/K⁺-ATPase的酶活开始随胁迫时间增长而上升;最后,各组实验盐度下第2和第6对鳃Na⁺/K⁺-ATPase的酶活下降并趋于稳定。另外,胁迫1 d时,各组实验盐度下三疣梭子蟹前5对鳃Na⁺/K⁺-ATPase的酶活显著低于后3对鳃Na⁺/K⁺-ATPase的酶活。三疣梭子蟹对盐度变化的调节可分为被动应激期(酶活力下降)、主动调节期(酶活力逐渐上升)和适应期(酶活力稳定);三疣梭子蟹后3对鳃是离子转运、渗透压调节的主要部位。     

Biochemical alterations induced by Zn and Cd individually or in combination in the serum of <Emphasis Type="Italic">Oreochromis niloticus</Emphasis>

Measurement of serum biochemical parameters in response to metal exposures can be especially useful to help identify target organs of toxicity as well as the general health status of animals. Oreochromis niloticus were exposed to 5.0 mg/L Zn, 1.0 mg/L Cd, and 5.0 mg/L Zn + 1.0 mg/L Cd mixture for 7 and 28 days, and alterations in serum enzyme activities and ion levels were measured. Significant changes in all the biochemical parameters were found to be time dependent. Following metal exposure, alkaline phosphatase activity was elevated at both exposure periods. No changes in activities of lactate dehydrogenase and lipase were observed in response to single or combined Zn and Cd exposure at 7 days while they increased at 28 days. Fish exposed to metals showed a decrease in cholinesterase activity at 7 days followed by a return to control levels at the end of the exposure period. The individual and combined effects of metals caused a decline in levels of Na⁺, Cl⁻, and Ca²⁺, especially at 28 days. K⁺ level increased at 7 days but it returned to control levels with increasing duration of exposure. This study indicated that the alterations in serum parameters may be a result of the target tissue (i.e., liver, gill, and kidney) damage and dysfunction induced by the metals and that these parameters can thus be used to assess the effects of metals on organisms.     

Biochemical and molecular differences in diploid and triploid ocean-type chinook salmon (<Emphasis Type="Italic">Oncorhynchus tshawytscha</Emphasis>) smolts

There is increasing evidence for complex dosage effects on gene expression, enzyme activity and phenotype resulting from induced ploidy change. In this study, ocean-type chinook salmon were bred using a 2 × 2 factorial mating design to create four families and test whether triploidization resulted in changes in growth performance and smolting. Eggs were pressure shocked after fertilization to create triploid fish from a subset of each family. In June, fish were sampled for size, plasma insulin-like growth factor 1 (IGF-1), gill Na⁺–K⁺-ATPase activity, and expression of two Na⁺–K⁺-ATPase α subunits in the gill. Diploids were significantly heavier than triploids, and there were significant differences due to family. Despite a significant positive correlation between plasma IGF-1 and fish size, plasma IGF-1 did not differ between diploid and triploid smolts. Diploids also had significantly greater gill Na⁺–K⁺-ATPase enzyme activities than triploids and there was a strong family effect. Gill Na⁺–K⁺-ATPase α1b isoform expression differed significantly by family, but not ploidy, and generally families with lower Na⁺–K⁺-ATPase enzyme activity had higher α1b isoform gene expression. Na⁺–K⁺-ATPase α1a isoform expression did not differ among any of the groups. Although diploids were larger and had higher specific activities of Na⁺–K⁺-ATPase in the gills, there was no difference in gene expression or circulating hormone levels. The strong family effect, however, suggests that strain selection may be useful in improving performance of triploids for aquaculture.     

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.     

Physiological characteristics and stress resistance of great sturgeon (<Emphasis Type="Italic">Huso huso</Emphasis>) juveniles fed with vitamins C,E, and HUFA-enriched <Emphasis Type="Italic">Artemia urmiana</Emphasis> nauplii

This study was carried out to examine the effect of Artemia urmiana nauplii enriched with HUFA, and vitamins C and E on stress tolerance, hematocrit, and biochemical parameters of great sturgeon, Huso huso juveniles. Cod liver oil (EPA 18% and DHA 12%), ascorbyl-6-palmitate and α-tocopherol acetate were used as lipid, and vitamin C and E sources, respectively. Beluga juveniles at the stage of first feeding (69.7 ± 5.9 mg body weight) were randomly divided into five treatments and three tanks were assigned to each diet. All fish groups were fed non-enriched Artemia for the initial 5 days and then fed enriched Artemia for 7 days. Juveniles were fed with Artemia enriched with HUFA + 20% vitamin C (C group); HUFA + 20% vitamin E-enriched Artemia nauplii (E group); HUFA + 20% vitamin C + 20% vitamin E (C and E group); HUFA without vitamins (HUFA) and non-enriched Artemia (control). After the period of enrichment, Juveniles were fed with Daphnia sp. from the 13th to the 40th day. At day 40, the fish were transferred directly from fresh water (0.5 ppt) to brackish water (6 ppt for 4 days and 12 ppt for 2 days) and warm water (from 27 to 33°C) to evaluate juvenile resistance to salinity and thermal shocks. Moreover, all treatments were separately exposed to freshwater in tanks with the same capacity as used for osmotic and thermal tests (as fresh water control). The addition of vitamins C, E, and C + E to HUFA significantly increased fish resistance to 12 ppt salinity and temperature stress tests, whereas survival was not significantly different among challenges at 6 ppt. There was no significant difference in the hematocrit index under stress conditions. Enrichment had significant influence on plasma Na⁺ level in the C group on the 4th day at 6 ppt. Na⁺ and Ca²⁺ concentrations in C, E, and C and E groups on the 1st day at 12 ppt, and Ca²⁺ level in E group on the 2nd day at 12 ppt were lower than the other groups. The glucose level in the C and C and E groups was lower than the other treatments on the 1st day at 12 ppt and the 2nd day at 33°C. Regardless of Artemia enrichment, plasma ions (Na⁺, K⁺, Ca²⁺, and Mg²⁺) and glucose concentrations in fish exposed to salinity stress tests were higher than fish in fresh water. Glucose concentration in plasma also increased after 2 days at 33°C. Although most of our results were not significantly different, the use of vitamins C, E, and HUFA in Artemia enrichment can improve Juveniles tolerance under stress conditions, and regardless of enrichment, these data show that beluga juveniles are partly sensitive to high salinity and temperature.     

Effects of Na/K and Mg/Ca ratios in saline groundwaters on Na-K-ATPase activity, survival and growth of Marsupenaeus japonicus postlarvae

The effects of the Na⁺/K⁺ and Mg²⁺/Ca²⁺ ratios in saline groundwaters on Na⁺-K⁺-ATPase activity, survival and growth of Marsupenaeus japonicus postlarvae were investigated. The results indicate that the Na⁺-K⁺-ATPase activity, survival rate and weight gain of postlarvae were significantly affected by the Na⁺/K⁺ and Mg²⁺/Ca²⁺ ratios (P < 0.05). The Na⁺-K⁺-ATPase activity of postlarvae, in every treatment, changed corresponding to Na⁺/K⁺ and Mg²⁺/Ca²⁺ ratios, and came to a stable level after 24 h. There was a negative relation between Na⁺-K⁺-ATPase activity and Na⁺/K⁺ ratio, while there was a positive relation between Na⁺-K⁺-ATPase activity and Mg²⁺/Ca²⁺ ratio. Compared with seawater (the Na⁺/K⁺ and Mg²⁺/Ca²⁺ ratios are 27.8 and 4.64 respectively), the Na⁺-K⁺-ATPase activity of the Na⁺/K⁺ ratio 30 treatment showed no significant difference, while the Mg²⁺/Ca²⁺ ratio 4.5 treatment showed distinct difference. The survival rates and weight gain of postlarvae increased markedly when the suitable amount of K⁺ and Ca²⁺ was added to test water, and arrived at their maximum in the Na⁺/K⁺ ratio 20-30 or Mg²⁺/Ca²⁺ ratio 4.5 treatment, having no significant difference compared with normal seawater. Therefore, considering the Na⁺/K⁺, Mg²⁺/Ca²⁺ ratios and the absolute concentration of Mg²⁺, Ca²⁺ in the experimental saline groundwaters applied to Marsupenaeus japonicus farming, it should be modulated to around 30, 4.5 and 1312 mg/l, 291 mg/l, respectively.     

Effect of a probiotic bacterium <Emphasis Type="Italic">Bacillus circulans</Emphasis> PB7 in the formulated diets: on growth,nutritional quality and immunity of <Emphasis Type="Italic">Catla catla</Emphasis> (Ham.)

Bacillus circulans PB7, isolated from the intestine of Catla catla, was evaluated for use as a probiotic supplement in the feeds for the fingerlings of Catla catla. The effect of supplement on the growth performance, feed utilization efficiency, and immune response was evaluated. Catla fingerlings (ave. wt. 6.48 ± 0.43 g) were fed diets supplemented with 2 × 10⁴ (feed C1), 2 × 10⁵ (feed C2), and 2 × 10⁶ (feed C3) B. circulans PB 7 cells per 100 g feed for 60 days at 5% of the body weight per day in two equal instalments in triplicate treatments. The control feed (CC) was not supplemented with the B. circulans. All the feeds were isocaloric and isonitrogenous. Fish fed with feed C2 displayed better growth, significantly (P ≤ 0.05) highest RNA/DNA ratio, a lower feed conversion ratio (FCR), and a higher protein efficiency ratio (PER) than the other experimental diets. Highest carcass protein and lipid was also observed in the fish fed C2 feed compared to the others. Significantly (P ≤ 0.05), highest protease was recorded in fish fed feed C2 (47.9 ± 0.016) and lowest in fish fed feed C3 (32.10 ± 0.009), where α-amylase activity did not differ significantly (P ≤ 0.05) beyond the lowest inclusion level. ALP, ACP, GOT, and GPT in the liver of Catla catla were the highest (P ≤ 0.05) in fish fed C2 feed. The highest TSP, albumin, and globulin was observed in fish treated with C2 feed after 60 days feeding trial, but the lowest glucose level was observed in the same treatment. After the feeding trial, the non-specific immunity levels and disease resistance of fish were also studied. Phagocytic ratio, phagocytic index, and leucocrit value were the highest in fish fed feed C2. After the feeding trial, the fish were challenged for 10 days by bath exposure to Aeromonas hydrophila (AH1) (10⁵c.f.u. ml⁻¹ for 1 h, and, after 7 days, 10⁷c.f.u. ml⁻¹ for 1 h). Highest survival percentage was observed in fish fed with feed C2 compared with only 6.66% in the controls, which indicated the effectiveness of B. circulans PB 7 in reducing disease caused by A. hydrophila.     

Use of vitamin C as an immunostimulant. Effect on growth,nutritional quality,and immune response of <Emphasis Type="Italic">Labeo rohita</Emphasis> (Ham.)

Teleost fish lack the enzyme for endogenous synthesis of ascorbic acid (AA), an essential micronutrient for fish. The aim of this study was to examine the effect of higher levels of dietary vitamin C on growth, nutritional quality, and immunomodulation in the Indian major carp, rohu (Labeo rohita). Four groups of L. rohita were fed experimental diets containing either no vitamin C (control) or supplemented with vitamin C at 500 mg kg⁻¹ (Exp-1), 1000 mg kg⁻¹ (Exp-2), or 1500 mg kg⁻¹ (Exp-3) for 60 days. Growth parameters (NWG, ADG, and SGR), serological parameters (TSP, TSA, TSG, and A:G), haematological parameters (TLC, TEC, Hct, MCV, and MCH), and different non-specific immunological parameters (PR, PI, respiratory burst activity, and bactericidal activity) were evaluated during the experimental trial. Fish fed a vitamin C-supplemented diet showed higher specific growth rate (SGR) up to 1000 mg kg⁻¹ compared with control fish. Different haematological and serological parameters along with non-specific immune parameters were influenced by vitamin C supplementation. Among the non-specific immune parameters phagocytic activity (PR and PI) and respiratory burst activity (NBT cells) were significantly (P ≤ 0.05) enhanced by increasing doses of vitamin C supplementation. Higher levels of dietary vitamin C significantly (P ≤ 0.05) enhanced protection against Aeromonas hydrophila (AH1) infection compared with controls. Results from this study help to establish the beneficial effect of vitamin C on growth and immunmodulation in rohu (L. rohita).     

Effects of triploidy on the Caspian salmon <Emphasis Type="Italic">Salmo trutta caspius</Emphasis> haematology

The aim of this study was a comparison of key haematological features of diploid (2n) and triploid (3n) Caspian salmon (Salmo trutta caspius). Morphometric indices of erythrocytes were determined on blood smears by light microscopy. Triploidy significantly (P < 0.001) increased all morphometric indices measured in the erythrocytes including cell size, cell surface area, and cell volume. The increase in cell size was larger for the major (27%) axis than for the minor (22%) axis, thus making erythrocytes of 3n Caspian salmon more ellipsoidal. The estimated increase in erythrocyte nuclear volume (87%) was bigger than the theoretical expected 50% increase. Haematological indices were measured manually by hemocytometry. Triploids had lower numbers of red blood cells (RBC: 1,120,000 cells/mL in 2n vs. 700,000 cells/mL in 3n; P < 0.001) but they were larger in size (mean erythrocytic volume [MEV]: 363.1 nm³ in 2n vs. 483.3 nm³ in 3n; P < 0.001). The decrease in RBC number was not compensated by the increase in MEV and, thus, triploidy affected the haematocrit (Hct: 38.8% in 2n vs. 33.06% in 3n; P < 0.05). Total blood hemoglobin concentration was lower in triploid fish (Hb: 9.9 g/dL in 2n vs. 8.9 g/dL in 3n; P < 0.05). In contrast, mean erythrocytic hemoglobin (MEH: 95 μg in 2n vs. 133.2 μg in 3n; P < 0.001) was higher for 3n Caspian salmon as a result of their larger erythrocytes, although MEH concentration (MEHC: 0.26 g/dL in 2n vs. 0.27 g/dL in 3n) did not significantly differ (P > 0.05). White blood cell (WBC) counts (lymphocytes and neutrophiles) were measured and WBC/RBC ratios were calculated. There were no significant differences in WBC (15,710 cells/mL in 2n vs. 12,683 cells/mL in 3n; P > 0.05), lymphocytes, and neutrophils as %WBC as well as WBC/RBC ratios between two ploidy levels (P > 0.05). Triploid Caspian salmon showed higher erythrocyte abnormalities such as ‘twisted’, ‘tailed’, and ‘anucleated’ cells as well as high portions of immature RBC in blood smears in comparison with diploids (P < 0.001).     

Enhanced hypoosmoregulatory response to growth hormone after cortisol treatment in immature rainbow trout,Salmo gairdneri

The growth-independent effect of ovine growth hormone (oGH) and oGH + cortisol treatment on seawater (SW) adaptation in immature rainbow trout, Salmo gairdneri was investigated. Fish were injected every second day with saline, 2.0 μg oGH/g or 2.0 μg oGH + 8.0 μg cortisol/g for a maximum of 8 injections in freshwater (FW). Subgroups were transferred to 28‰ SW after 4 or 8 injections, and changes in plasma Na⁺ and Cl⁻, muscle water content and gill Na⁺/K⁺-ATPase activity were measured. In both of the hormone-treated groups retained in FW, gill Na⁺/K⁺-ATPase activity and interlamellar chloride cell density increased. The effects were most pronounced in the oGH + cortisol group after 2 weeks of treatment. After transfer to SW most of the control fish died due to the osmotic stress, whereas in the hormone-treated groups, mortality was low and there was a positive correlation between pretransfer gill Na⁺/K⁺-ATPase and the ability to maintain ionic-osmotic homeostasis after SW transfer. After two weeks of oGH + cortisol treatment, gill Na⁺/K⁺-ATPase activity was maximal. In contrast, after SW transfer, Na⁺/K⁺-ATPase activity increased further in the oGH-treated group. This group regulated ionic-osmotic parameters less effectively than the oGH + cortisol-treated group. The data indicate that GH and cortisol are important hormones in the regulation of hypoosmoregulatory mechanisms in S. gairdneri.     

Effect of prebiotic xylooligosaccharides on growth performances and digestive enzyme activities of allogynogenetic crucian carp (<Emphasis Type="Italic">Carassius auratus gibelio</Emphasis>)

The effect of prebiotic xylooligosaccharides (XOS) on the growth performance and digestive enzyme activities of the allogynogenetic crucian carp, Carassius auratus gibelio, was investigated. XOS was added to fish basal semi-purified diets at three concentrations by dry feed weight: diet 1, 50 mg kg⁻¹; diet 2, 100 mg kg⁻¹; diet 3, 200 mg kg⁻¹, respectively. Twelve aquaria (n = 20) with three replicates for each treatment group (diets 1–3) and control treated without XOS were used. Weights of all collected carp from each aquarium were determined at the initial phase and at the end of the experiment, and the carp survival was also determined by counting the individuals in each aquarium. After 45 days, there were significant differences (P < 0.05) in the relative gain rate (RGR), and daily weight gain (DWG) of diets 1–3 were compared with the control. However, the survival rate was not affected (P > 0.05) by the dietary treatments. For enzymatic analysis, dissection produced a crude mixture of intestine and hepatopancreas of each segment to measure. The protease activity in the intestine and hepatopancreas content of fish in diet 2 (487.37 ± 20.58 U g⁻¹ and 20.52 ± 1.93 U g⁻¹) were significantly different (P < 0.05) from that in the control (428.13 ± 23.26 U g⁻¹ and 12.81 ± 1.52 U g⁻¹) and diet 3 (428.00 ± 23.78 U g⁻¹ and 14.04 ± 1.59 U g⁻¹). Amylase activity in the intestine was significantly higher for diet 2 compared to diet 1 and the control. As for amylase in the hepatopancreas, assays showed higher activity in diet 2 (P < 0.05) compared to the rest.     

Effects of sulfide on K+ flux pathways in red blood cells of crucian carp and rainbow trout

The effect of sulfide on K⁺ influx pathways was measured in red blood cells (RBCs) of sulfide-sensitive rainbow trout (Oncorhynchus mykiss) and sulfide-tolerant crucian carp (Carassius carassius). In trout RBCs, maximal inhibition of Na⁺, K⁺-ATPase was attained at 10 mol l^–1 sulfide and amounted to 32% without being influenced by pH between 6.7 and 8.3. Ouabain-resistant K⁺ influx in the absence and presence of sulfide was insignificant at pH values between 6.7 and 7.7. At higher pH values ouabain-resistant K⁺ influx increased, but was inhibited to about 15% by 30 mol l^–1 sulfide. In RBCs of crucian carp neither Na⁺, K⁺-ATPase nor ouabain-resistant K⁺ influx were affected by sulfide concentrations up to 850 mol l^–1. Differences in sulfide-sensitivity of K⁺ influx between both species can be based upon different properties of the membrane transporter themselves. The reduced Na⁺, K⁺-ATPase activity in trout RBCs may also result from a slightly reduced (by 9%) ATP level after sulfide exposure. In addition, intracellular sulfide concentrations were higher in trout RBCs as compared to crucian carp. In trout, intracellular sulfide concentrations reached extracellular levels within 5 min of incubation whereas sulfide concentrations in crucian carp RBCs remained about 2-fold lower than extracellular concentrations. Although the physiological basis of sulfide-insensitive K⁺ influx in crucian carp RBCs is currently unknown it may contribute to the extremely high sulfide-tolerance of this species.     

壳聚糖对Cd2+和Pb2+的吸附作用

为开发新型吸附剂,降低重金属对水体的污染,研究了壳聚糖对Cd²⁺和Pb²⁺的吸附条件,探讨了pH、温度、反应时间、壳聚糖添加量和金属离子初始浓度等因素对壳聚糖吸附性能的影响。结果表明,pH 7~8和pH 5~6条件下壳聚糖对Cd²⁺和Pb²⁺的吸附能力最强;低温有利于壳聚糖的吸附;在8 h时壳聚糖对Cd²⁺的吸附容量达到最大,而对Pb²⁺的吸附在实验时间内是随着吸附时间的延长而增大;随着壳聚糖添加量的增加,其对Cd²⁺和Pb²⁺的吸附能力也增强;初始金属离子浓度的变化对Cd²⁺的影响不大,而在高的金属浓度下对Pb²⁺的吸附率显著降低。壳聚糖对Cd²⁺、Pb²⁺的吸附动力学和热力学分别符合Lagergren方程二级吸附模型和Langmuir吸附方程。研究表明,壳聚糖对不同金属离子的吸附能力不同,在单一金属溶液中,壳聚糖对Cd²⁺的吸附能力要强于对Pb²⁺的吸附能力。系统地研究了壳聚糖对Cd²⁺和Pb²⁺的吸附条件及性能,为壳聚糖作为重金属吸附剂的应用提供理论依据。