Expression of HSP70 in response to heat-shock and its cDNA cloning from Mediterranean blue mussel

Expression of HSP70 in response to heat-shock was investigated at the protein and mRNA levels in Mediterranean blue mussel. Western and Northern blot analyses revealed that HSP70 was expressed following heat-shock in the mantle at both protein and mRNA levels, suggesting that gene expression of HSP70 is implicated in the cellular response to heat-shock stress in mussel. It was then attempted to clone HSP70 cDNA in order to determine the primary structure of mussel HSP70. As a result, two full-length cDNA encoding HSP70 were isolated from a cDNA library prepared from the heat-shocked mantle. The isolated cDNA consist of single open reading frames of 2067 bp and 1911 bp which encode proteins of 689 amino acids and 637 amino acids, respectively. Both HSP70 cDNA encode an ATPase do main, and a substrate-binding do main in addition to a Glu-Glu-Val-Asp (EEVD) peptide motif that is specific for cytosolic HSP70. These findings suggest that the cDNA clones obtained in the present study encode cytosolic HSP70.     

Age estimation of the wild population of Japanese mantis shrimp <Emphasis Type="Italic">Oratosquilla oratoria</Emphasis> (Crustacea: Stomatopoda) in Tokyo Bay,Japan, using lipofuscin as an age marker

ABSTRACT:   In this study, the age composition of the Japanese mantis shrimp, Oratosquilla oratoria , in Tokyo Bay, central Japan, was investigated using lipofuscin, an autofluorescent pigment, as an age marker. Lipofuscin in a histological section of protocerebral bridge cell mass (PBCM) in the brain was identified by confocal microscopy, and its concentration was quantified by image analysis. Modal analysis of the lipofuscin concentration showed four regularly-spaced modes that could each be regarded as a distinct age group. This implied a constant lipofuscin accumulation in PBCM at a 6.5 × 10⁻²% volume fraction per year; it also implied the existence of individuals that are at least 4 years old. The lipofuscin concentration was found to be a more appropriate index than body length for estimating the age of O. oratoria , because the modal analysis on the body–length histogram failed to detect apparent age groups. This was probably a result of the declining growth rate with age and the individual variations in growth through molting leading to considerable overlap in the size between different age groups. The lipofuscin analysis suggested that fast-growing individuals in each cohort are recruited to the fishery, and most individuals attain the exploitable size between 2 and 3 years of age.     

The effects of vitamin E on non-specific immune response of the juvenile soft-shelled turtle <Emphasis Type="Italic">Pelodiscus sinensis</Emphasis>

ABSTRACT:   This study was conducted to determine the effect of dietary vitamin E supplements on non-specific immune responses in juvenile soft-shelled turtles. Turtles were fed diets supplemented with vitamin E at 0 (control), 50, 250, 500, 1000 and 5000 mg/kg, respectively, for 4 weeks. Results showed that blood cell phagocytosis and serum bactericidal activity were significantly improved in turtles from two diets (250 and 500 mg/kg vitamin E supplementation), while had no significant improvement in turtles from the three remaining diets (50, 1000 and 5000 mg/kg vitamin E supplementation) when compared to turtles from the control group. Serum bacteriolytic activity in turtles from diets with 50, 250, 500 and 1000 mg/kg vitamin E supplementation were higher than activity levels quantified for the control group, but no significant difference was observed between the 5000 mg/kg vitamin E supplementation diet and the control diet. The results suggest that vitamin E has an upper and lower threshold for improving non-specific immune function in soft-shelled turtles, and the optimal supplementation may be between 250 and 500 mg/kg.     

Acute tolerance and histopathological effects of ammonia on juvenile maroon clownfish Premnas biaculeatus (Block 1790)

This study evaluated the median lethal concentration values (LC₅₀) and the histopathological effects of un‐ionized ammonia (NH₃^‐‐N) on juvenile maroon clownfish Premnas biaculeatus. After 96 h of exposure to different concentrations of ammonia, juveniles were sampled for histopathological evaluation. The 24 and 96 h LC50 values of NH₃^?‐N determined were 1.68 and 0.89 mg L^?1 respectively. Maroon clownfish exposed to different ammonia concentrations displayed histopathological alterations in the gills, kidney, liver and brain. Gill tissue damage included lamellar hyperplasia, lamellar shorting and hyperplasia and hypertrophy of mucous cells. The kidney showed hyperanaemia, enlarged sinusoids within an apparently decreased amount of haematopoietic tissue, oedema on tubular cells and tubular necrosis, and an enlarged Bowman's capsule. The liver presented dilatation of hepatic sinusoids, fatty deposition in hepatocytes and Mallory bodies. Examination of the brain revealed a proliferation of glial cells, and the Virchow‐Robin space indicated a severe perivascular oedema and signs of neuronal suffering with satellitosis. The results of this study indicate that juvenile maroon clownfish are relatively sensitive to ammonia and particular attention must be given to this toxic compound in culture systems.     

Measurements of auditory sensitivity in common carp <Emphasis Type="Italic">Cyprinus carpio</Emphasis> by the auditory brainstem response technique and cardiac conditioning method

ABSTRACT:   Recently, the non-invasive auditory brainstem response (ABR) technique has been applied to the determination of hearing sensitivity in fish. The technique has some advantages over, and methodological differences from, the classical techniques. The auditory thresholds of common carp were measured by the ABR technique. The classical method was applied to measure cardiac response by electrocardiogram (ECG) after conditioning with electric shock. The most sensitive frequency obtained by ABR was 505 Hz, and by ECG, 1000 Hz. The shapes of the audiogram obtained by ABR and ECG were similar, though threshold levels in both audiograms differed from each other. Furthermore, audiograms obtained by using behavioral techniques were compared with those taken by the ABR and ECG techniques to assess the validity of the ABR technique for testing fish hearing sensitivity. The measured threshold levels increased in the order of behavioral, ABR, and ECG techniques, in the range below 1000 Hz.     

三沙湾海域化学需氧量（COD）分布特征及相关性分析

2012年5月和11月对三沙湾海域水质化学需氧量( COD)进行大面积调查研究,结果表明,2012年三沙湾海域COD含量变化范围为0．52~2．13 mg/L,平均含量为0．94 mg/L;水平分布特征呈西北侧向东南水域逐渐递减的趋势,白马港海域COD含量最高;秋季高于春季;平均COD含量能达到一类海水水质标准要求;化学需氧量与活性硅酸盐、无机氮、总氮及悬浮物呈极显著正相关;与pH值、盐度呈极显著负相关。     

The effects of less-expensive plant protein sources replaced with soybean meal in the juvenile diet of grass carp (Ctenopharyngodon idella): growth, nutrient utilization and body composition

In this study, the effects of less-expensive protein sources replaced with soybean meal on growth performance, nutrient utilization and body composition of juvenile grass carp were investigated. For this purpose, a control diet containing 42% soybean meal was prepared. Cotton seed meal (CSM), sunflower meal (SFM) and corn meal (CM) were added to the experimental diets by replacing 100% of the soybean meal protein used in the control diet. All diets were iso-nitrogenous (40% crude protein) and iso-caloric (19.5?MJ?kg^?1 gross energy). Each treatment had three replicates, and 20 juvenile grass carps (3.5?±?0.1?g, initial weight) were located in per replicate. The fish were fed to satiety three times in a day during 3?months at 26°C of water temperature in glass aquarium (45 l). The results of the study showed that the weight gain (308.6?C448.6%), specific growth rate (1.56?C1.89%/day), feed conversion rate (1.2?C1.9), protein efficiency ratio (1.32?C2.08), and apparent digestibility coefficients of protein (80.5?C89%) or sum of amino acids (80.5?C88.9%) were not significantly affected by the experimental diets, except diet CM. All of these values obtained for the diet CM were significantly lower than those of fish fed on the control, CSM and SFM diets. However, there were no significant differences between the lipid digestibility of the diets, and crude protein, lipid, and gross energy contents in the whole body of fish in experiments. In conclusion, CSM and SFM diets were highly utilized by juvenile grass carp, but not CM diet. These results combined with the remarkable acceptability of diets containing high levels of plant protein ingredients with identical growth performances of juvenile grass carp show clearly that dietary soybean meal level can be considerably reduced without any adverse consequence in terms of somatic growth and nitrogen utilization.     

Effect of the dusk photoperiod change from light to dark on the incubation period of eggs of the spotted rose snapper, Lutjanus guttatus (Steindachner)

Spotted rose snapper, Lutjanus guttatus (Steindachner), eggs were incubated under different photoperiods to examine the effect of photoperiod on incubation. The eggs from two fish were incubated under five artificial photoperiods: constant dark (D), constant light (L) from 06:00 hours and 6, 10 and 14 h of light from 06:00 hours. The eggs from seven other fish were incubated under a natural photoperiod. Different spawning times (21:00 – 01:00 hours) and different photoperiods combined to give the start of the dusk photoperiod change after 11–23 h of incubation. Constant light or applying the dusk photoperiod change after ≥20 h of incubation appeared to extend the hatching period. The mean hatching period for groups of eggs incubated in darkness or that received the dusk photoperiod change after ≤19 h of incubation (n=8 different groups) was 2 h 15±10 min, which was significantly lower (P<0.05) than the mean hatching period of 4 h±37 min for groups that did not receive the dusk photoperiod change or that received the dusk photoperiod change after ≥20 h of incubation (n=9 groups). However, despite these differences, the majority of the eggs hatched during a 2–3 h period from 17 to 20 h of incubation, and a sigmoid regression (r²=0.9) explained the relationship between percentage hatch and hours of incubation for all photoperiod groups.     

Effects of density on growth and survival of juvenile Red Swamp Crayfish, Procambarus clarkii (Girard), reared under laboratory conditions

The aim of this study was to quantify the importance of population density on the growth and survival of juvenile red swamp crayfish, Procambarus clarkii (Girard), and to understand the effects of density on both the period between moults and the increment per moult at high densities without food limitation. A laboratory experiment was performed to determine growth and survival of juvenile crayfish at five densities (20, 40, 60, 80 and 100 crayfish m^?2). Density clearly affected crayfish growth. The impact of density on final weight was higher than on total length increase, growth rate and final length (FL) (by decreasing order of effect). When comparing minimum and maximum densities, we observed that FL suffered a 34% reduction. Number of moults, mean intermoult period (IP) and survival were not significantly affected by density. Our results indicated that the IP and the percentage of increment per moult are affected by the size of crayfish before moult and we provide equations for these relations. Our findings are relevant both for crayfish aquaculture management and for the management of wild or harvested rice‐field crayfish populations.