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).     

The effect of temperature on the energy budget of the Manila clam, <Emphasis Type="Italic">Ruditapes philippinarum</Emphasis>

In this study, the energy budget of the Manila clam, Ruditapes philippinarum, was evaluated after one-week acclimation periods at 5, 10, 15, 20, and 25°C. Small clams (151 ± 12 mg DW) and large clams (353 ± 16 mg DW) were fed with the microalgae, Isochrysis galbana. Filtration rate, ingestion rate, assimilation efficiency, oxygen-consumption rate, and ammonia excretion rate were measured. Both filtration rate and ingestion rate of small and large clams were found to be related to temperature. The highest Q ₁₀ values were measured in the range 15–20°C for both small and large clams. Assimilation efficiency of both small and large clams was not significantly influenced by temperature, although the maximum mean values were detected at 20°C. Oxygen consumption rate and ammonia excretion rate of small and large clams were found to be related directly to temperature over the entire range, with a maximum being detected at 25°C. The highest Q ₁₀ value was estimated in the range 10–15°C with regard to oxygen consumption rate, and in the range of 15–20°C with regard to ammonia excretion rate. Scope for growth (SFG) was positive at all temperatures, achieving a maximum value at 20°C in both small and large clams, primarily as a consequence of the enhanced ingestion rate which offset the concomitant elevation in the metabolic rate. In this study we have estimated the thermal optimum for this species at 20°C.     

Growth,absorption and assimilation efficiency by mature cuttlefish (<Emphasis Type="Italic">Sepia officinalis</Emphasis>) fed with alternative and artificial diets

The effects of feeding three natural frozen diets, grass shrimp (Palaemonetes sp.), crayfish (Procambarus clarkii) and fish (Sardina pilchardus) and two semi-humid artificial diets (based on fish powder) to mature cuttlefish, Sepia officinalis, were analysed. Growth and feeding rates (GR and FR, % BW day⁻¹), food conversions (FC, %), and total protein and lipid composition of the diets were determined. Digestive gland to body weight ratio and absorption efficiency were calculated for each diet. Cuttlefish fed shrimp and crayfish grew larger (1.5 and 1.1% BW day⁻¹, respectively) compared to the other diets. Shrimp promoted the highest FC, followed by crayfish, and sardine. The highest FR was obtained for cuttlefish fed crayfish (8.4% BW day⁻¹). Although both artificial diets were accepted, none produced growth. A positive correlation (r = 0.96) between cuttlefish ingestion rate and digestive gland weight was obtained. Some cannibalism occurred among cuttlefish fed the artificial diets during the last week of the experiment. According to the results obtained, P. clarkii could be used as an alternative prey to shrimp for rearing adult mature (>50 g) S. officinalis.     

Differential HSP90α expression in fish hepatocytes from polluted estuary during summer

The molecular chaperone, heat shock protein 90α (HSP90α), plays an important role in protein folding, degradation of denatured proteins and steroid activation. It is essential for the maintenance of cellular integrity and survival when induced in response to environmental, physical and chemical stresses. In the present investigation the effect of environmental stress on HSP90α expression was examined in grey mullet Mugil cephalus living in either a contaminated (Ennore) or uncontaminated (Kovalam) estuary over two seasons: Hepatocytes were isolated from grey mullet of both estuaries. Oxidative stress was determined along with HSP90α in these fish. Additionally, immunohistochemical changes were studied to confirm the HSP90α expression. Comparison of the results revealed enhanced hepatocyte oxidative stress and HSP90α expressions in fish from Ennore to a significant extent than fish from Kovalam. Also, the results showed significant seasonal variations with maximum expression observed during summer compared to the monsoon season. Overexpression of HSP90α in hepatocytes exposed to chronic environmental stress by pollutants may confer differential effects on cell survival by protecting against oxidative stress induced changes. The results also indicate that seasonal variations have significant effect on the HSP90α expression.     

Application of ELISA-based kit for detecting AOZ and determining its clearance in eel tissues

Furazolidone, an antibacterial drug that was once widely used in the livestock industry and aquaculture, is now prohibited in numerous countries. It is difficult to detect residual furazolidone because it is readily metabolized in animal tissues but, by using and liquid chromatography coupled with tandem mass spectrometry, its metabolite, 3-amino-2-oxazolidinone (AOZ) can be detected. Here we describe the validity of an enzyme-linked immunosorbent assay (ELISA) kit to detect AOZ in Japanese eel Anguilla japonica tissue. ELISA is capable of detecting AOZ at 1.0 μg/kg in an eel sample with excellent accuracy and precision. Our results show that ELISA is suitable for regulatory purposes and for studying the fate of AOZ residues in eel treated with furazolidone. To measure the persistence of AOZ in eel tissues, eels (1.4–6.5g) were immersed in tanks containing 2 and 10 mg furazolidone/L for 3 h, and then maintained in a tank supplying well water for the next 160 days. The half-lives of AOZ, calculated from the linear terminal part of the excretion curve, were 25.0 days in muscle and 21.6 days in liver from fish exposed to 2 mg/L furazolidone. In the eels treated with 10 mg/L furazolidone, by contrast, high levels of AOZ were detected in liver and muscle, but the half-lives of AOZ were similar to those in fish treated with 2 mg/L furazolidone. The half-lives of AOZ in eel tissues were prolonged by the condition of low water temperature.     

Ecology,physiology and genetics of a phycodnavirus infecting the noxious bloom-forming raphidophyte <Emphasis Type="Italic">Heterosigma akashiwo</Emphasis>

Abstract:   Heterosigma akashiwo virus (HaV) is a large icosahedral virus (∼0.2 μm) harboring a double-stranded DNA (dsDNA) genome (∼294 kbp). The virus is the only member of the genus Raphidovirus in the family Phycodnaviridae. Since its first discovery, a number of ecologic, physiologic and genetic studies about HaV have been conducted; especially, the relationship between H. akashiwo and HaV in nature was studied and viral infection is now regarded as a significant factor influencing the dynamics and termination of H. akashiwo blooms. HaV infection has considerable impacts on H. akashiwo populations in both aspects of fluctuation in biomass (quantity) and changes in clonal composition (quality). Partial sequencing of the HaV genome revealed that a number of genes showed considerable similarity to those of other protist-infecting viruses; still, the phylogenetic position of HaV suggested a number of enigmas in host–virus coevolution. Here are summarized the ecology, physiology and genetics of HaV especially from the viewpoint of the host–virus relationship.     

Induced thermotolerance and expression of heat shock protein 70 in sea cucumber <Emphasis Type="Italic">Apostichopus japonicus</Emphasis>

ABSTRACT:   Thermal limits, induced thermotolerance and the expression of heat shock protein 70 (Hsp70) in an echinoderm Apostichopus japonicus were studied. The sublethal and lethal temperatures for the juveniles were 30 and 34°C, respectively; a previous sublethal heat shock exposure (30°C, 2 h) could increase the survival rates of the sea cucumbers when they were exposed to 34°C. This induced thermotolerance could last for at least 2 days. Levels of Hsp70 increased substantially after sublethal heat shock exposure and linearly decreased with time. This result indicated that a close relationship existed between the induction of thermotolerance and the levels of Hsp70 in A. japonicus.     

Effects of silvering state on induced maturation and spawning in wild female Japanese eel <Emphasis Type="Italic">Anguilla japonica</Emphasis>

ABSTRACT:   The effects of silvering state of wild female Japanese eels Anguilla japonica on the success of induced maturation and the following spawning were examined. Thirty-eight females, collected in Mikawa Bay, were divided into four stages based on their silvering state: yellow (Y1), late-yellow (Y2), silver (S1) and late silver eels (S2). Despite injections of salmon pituitary extract (SPE) through the standard technique, Y1 and Y2 eels did not respond to the treatment with undeveloped gonad (gonad-somatic index [GSI]: 0.3–0.9), and all these females died by 5 weeks, probably due to an abnormal physiological condition. Most S1 (81%) and S2 eels (100%) matured completely (GSI: 17.8–51.4), and finally spawned successfully (69% for S1, 89% for S2). S2 eels fully matured with oocytes of over 750 μm in diameter by significantly smaller number of injections of SPE (5–6 times) than the case of S1 eels (6–8 times). The amount of eggs released by S2 eels (0.65 ± 0.11 g/fish per body weight [BW]) was significantly larger than those by S1 eels (0.54 ± 0.09 g/fish per BW). There was no difference in fertilization and hatching rates between eggs released by S1 eels and those of S2 eels. These results indicate that the success of induced maturation and spawning in wild female Japanese eels depends on their silvering state, and matured eggs can be obtained efficiently through the use of S2 eels rather than other stages.     

Carbonic anhydrase and Na/K-ATPase activities at different molting stages of the giant freshwater prawn <Emphasis Type="Italic">Macrobrachium rosenbergii</Emphasis>

ABSTRACT:   In the present study, the role of carbonic anhydrase (CA) and Na/K-ATPase in the gill and epidermal tissues in the giant freshwater prawn Macrobrachium rosenbergii was examined as a function of molting stage. CA activity levels in the front and back gills were low at the intermolt stage C₀, but increased significantly at premolt stage D₃, and then decreased after molting. In the epidermal tissue, activity levels decreased gradually towards premolt to a minimum level at stage D₃, but became elevated at postmolt stages A and B. Na/K-ATPase levels in the front and back gills did not change significantly during the molt cycle. CA in the gill is possibly involved in supplying counter-ions for ion uptake, while CA in the epidermal tissue may play a role in mineralizing the exoskeleton after ecdysis. Na/K-ATPase in the gills may function in ion uptake from the ambient medium; however, since its activity was not influenced by the molt cycle, it probably does not have a major role in osmoregulation in the freshwater environment.