Bioaccumulation of cadmium and its biochemical effect on selected tissues of the catfish (<Emphasis Type="Italic">Clarias gariepinus</Emphasis>)

The present study examines the pattern of accumulation of cadmium (Cd) and its biochemical effects on selected tissues of a variety of African catfish (Clarias gariepinus), after exposure to various doses of Cd. The results obtained indicate that at the end of 21 days of exposure, the total tissue organ cadmium concentration followed the pattern kidney > gill > liver > muscle for each of the exposure concentrations. The levels of Cd in these organs were higher than those in ambient water. Moreover, while the rate of uptake of Cd increased with time in the kidney, liver and muscle, it decreased in the gill. Superoxide dismutase (SOD) was significantly elevated only in the kidney of catfish treated with 0.2 and 0.4 ppm of Cd for 7 days compared with the control. Conversely, gill SOD was significantly decreased in the same concentrations of Cd-treated catfish relative to the control. Statistically similar levels of SOD were observed in the liver, brain and muscle with all the treatments after the same duration of treatment. In the fish exposed for 21 days, SOD activity was significantly decreased in the kidney with a corresponding increase in lipid peroxidation (LPO), but it manifested only with the 0.2- and 0.4-ppm Cd treatment relative to the control. In the liver, however, Cd exposure significantly increased SOD in the 0.2- and 0.4-ppm treatments of the same duration. In conclusion, the present study indicates that the accumulation of Cd and its effect on SOD and LPO in C. gariepinus is dependent on concentration, tissue and time     

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.     

Phospholipase A<Subscript>1</Subscript> activity of crude enzyme extracted from the ovaries of skipjack tuna

The phospholipid class composition, fatty acid composition and phospholipase A₁ (PLA₁) activity from the ovaries of skipjack tuna were compared with those of six other species of marine fish. In the skipjack ovaries, the lysophosphatidylcholine (LPC) proportion for the phospholipid, the docosahexaenoic acid (DHA) percentage for the total fatty acids of the phospholipids and the PLA₁ activity of the crude enzyme were the highest among those of the seven species. The optimum pH and temperature for the PLA₁ activity of the crude enzyme from the skipjack ovaries were in the range of pH 6–7 and 20–30°C, respectively, and calcium ions were not required. As a substrate, phosphatidylcholine was more easily hydrolyzed than phosphatidylethanolamine by this enzyme, and the plasmalogen-type phospholipid was much lower than the acyl-type phospholipid. After a 6-h hydrolysis reaction of the purified phospholipid extracted from the mixed ovaries of skipjack and yellowfin tuna by this enzyme, the LPC ratio of the phospholipid increased from 20 to 72.6% and the percentage of DHA for the total fatty acids of the phospholipid also increased. Thus, skipjack ovaries might possibly be used as a source of PLA₁.     

Reproductive season and female maturity size of coconut crab <Emphasis Type="Italic">Birgus latro</Emphasis> on Hatoma Island,southern Japan

Information about reproductive season, size at sexual maturity and size-dependent reproductive potentials is important for efficient resource management. Such information is necessary to determine seasonal closures and minimum legal size limits. To conserve resources of coconut crab Birgus latro, which are in danger of extinction due to excessive exploitation, their reproductive season, female size at functional maturity and relationship between female size and reproductive potentials were investigated. The reproductive season started in early June and ended late August, and females with higher reproductive potentials showed a tendency to spawn early in the reproductive season. The prohibition of exploiting the resources during the reproductive season, especially early in the season, would be appropriate for sustainability of high reproductive outputs. Fifty percent of investigated females matured functionally at 24.5-mm thoracic length (TL). However, the size at which all females reached functional maturity was 32.3 mm TL, and there was a wide variation in the size of functional maturity among individuals. We must determine minimum legal size by taking the variation into account to allow all individuals to mate at least once before they are caught, which may prevent serious depression in reproductive rate and genetic diversity.     

Direct evidence of multiple paternities in natural population of viviparous Japanese surfperch by allelic markers of microsatellite DNA loci

This study was performed to obtain information on the occurrence of multiple paternities in three species of viviparous Japanese surfperch using allelic markers of microsatellite DNA loci. Direct evidence for multiple fertilizations was established by reconstructing paternal genotypes from the progeny of gravid females. Multiple paternities were ascertained in five of 10 broods of Ditrema temmincki and in three of nine broods of Neoditrema ransonneti, but not in Ditrema viride. The number of patrilines detected in the progeny of D. temmincki and N. ransonneti females were two or three, respectively, as determined by the GERUD v2.0 algorithm for reconstructing parental genotypes from half-sib progeny arrays.     

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).     

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.