Molecular characterization of microalgae used in aquaculture with biotechnology potential

Microalgae are the main component of first tropic level in aquatic food chain; it is for this reason that they are used as food in aquaculture. Also due to its biotechnological potential properties, they are used in the production of diverse components, dyes, antioxidants, enzymes, emulsifiers, etc. The extended ways of microalgae applications require physiologically and genetically stable cultures as well as correctly identified organisms to guarantee reproducibility and reliability. But the variety of species and the morphological similarity between some of them make difficult the identification of some microalgae. The use of molecular markers has supplied a very useful tool for identification of microalgae in fast mode, such as in classification. The present study has worked on the molecular characterization of main species of microalgae used in aquaculture in base of the molecular markers 18S rRNA and 16S rRNA. Microalgae DNA has been amplified and sequenced, and the resultant sequences were analyzed and reflected in phylogenetic trees. The phylogenetic analyses obtained reflect as both molecular markers allow to differentiate the main genus used in aquaculture.     

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.     

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.     

Gonadal sex differentiation and sex control in red sea bream, Pagrus major

The gonadal sex differentiation in red sea bream, Pagrus major, which is one of the most important species for aquaculture in Japan, was revealed histologically. The suitable conditions for induction of all-male groups in the fish were investigated, and functional males were induced by the conditions of oral administration of 17α-methyltestosterone. The sex determination of this fish was also discussed.     

The relationship between water intake rates,paddy ponding depth,and farmers’ water management techniques

Japanese farmers manage their irrigation water based on their past experiences and preferences, considering such factors as weather and available water (hereafter defined as empirical water management). They elaborately control the intake and drainage rates of their own paddy fields to maintain optimal ponding depths. But these well-managed systems will drastically change because of the decreasing number of farmers. Therefore, it is necessary to clarify if the optimal ponding depth will be maintained within the limits of traditionally-allowed water intake rate from the main river. The first objective of this study was the quantification of actual water use in the paddy fields, resulting from the farmers water management on the basis of their experience. The significance of the present water intake rate under empirical water management was studied for a paddy field command area of about 230 ha. Water intake rates and the water requirements of the whole area were investigated by measuring the flow rate at 17 points of irrigation and drainage canals. Characteristics of the farmers empirical water management were investigated by measuring the hourly changes in inflow and outflow rates for a sub-area using an automatic measurement system, and an inferential method of determining water management patterns for the paddy fields was proposed. The newly-proposed inferential method was introduced in the tank model, which expresses the characteristics of water management in the command area. The Shuffled Complex Evolution Algorithm (SCE-UA) method was used for optimizing the model parameters. It was proven that the model accuracy improved when the farmers empirical water management was taken into account. The optimal amount of water to be applied to the command area was quantified by the simulation. The second objective was to predict the effect of the decreasing number of farmers on future water use conditions. The simulated result indicates the difficulty of maintaining optimal ponding depth for the whole command area when the farmers empirical water management is not maintained. In other words, results indicated that efficient water use requires an automatic water management system or a new pipeline system to replace the farmers present empirical water management.