An effective and practical method of net settings in rearing tank to suppress hypermelanosis in Japanese flounder

In Japanese flounder aquaculture, hypermelanosis occurs widely on the blind side. Rearing flounders in a net-lined tank was recently reported to prevent hypermelanosis. To effectively apply this method to larger tanks for aquaculture farming, the net setting method was examined. Juvenile flounders without darkened areas on the blind side [total length (TL) 13 cm] were selected, and reared for 6 months (TL 32 cm). In the control tank without a net, the median value of darkened area ratio (darkened area to blind side area) was 46%. By only covering the tank bottom with slack net, the darkened area ratio was suppressed to 8%, less than 1/5 of that in the control tank. At the end of experiment, bottom coverage ratios of 0%, 10%, 30%, 50%, and 100% revealed a negative correlation between bottom net coverage and darkened area ratio. In this experiment, the darkening area in the tank with 50% bottom net coverage decreased to 1/3 of the control. Although the occurrence of hypermelanosis differs depending on the production lot, these results are expected to serve as a reference for selecting the suitable net size to meet the level of clearness of the blind side.

     

Metabolism of fungicide diethofencarb in grape (Vitis vinifera L.): definitive identification of thiolactic acid conjugated metabolites

The metabolic fate of diethofencarb (isopropyl 3,4-diethoxycarbanilate) separately labeled with (14)C at the phenyl ring and 2-position of the isopropyl moiety was studied in grape (Vitis vinifera L.). The acetonitrile solution of (14)C-diethofencarb at a rate of 500 g a.i. ha(-)(1) was once applied topically to fruits or leaves at the maturity stage of fruits (PHI 35 days), and the plants were grown in the greenhouse until harvest. In the grape plants, diethofencarb was scarcely translocated to the untreated portion and was degraded more in the fruit as compared to the leaf. For the fruit, diethofencarb primary underwent O-deethylation at the 4-position of the phenyl ring to form the phenolic derivative, isopropyl 3-ethoxy-4-hydroxycarbanilate (0.9% of the total radioactive residue, TRR). This metabolite was successively transformed via conjugation with glucose at the phenolic hydroxy group (8.1-18.1% TRR) or with thiolactic acid at the 5-position of the phenyl ring (1.5-1.7% TRR). The thiolactic acid conjugate was further metabolized mainly to two different types of glucose conjugates at the 4-position of the phenyl ring (8.7-13.5% TRR) and the hydroxy group in the thiolactic acid moiety (6.4-7.3% TRR), as evidenced by (1)H NMR and atmospheric pressure chemical ionization-liquid chromatography-mass spectrometry together with cochromatographies with synthetic standards.     

Characterization of Treatment Processes and Mechanisms of COD, Phosphorus and Nitrogen Removal in a Multi-Soil-Layering System

Characteristics of the treatment processes inside a MSL system were investigated by using a laboratory-scale MSL system, which was set up in a D 10 × W 50 × H 73 cm acrylic box enclosing "soil mixture blocks" alternating with permeable zeolite layers. For the study of the treatment processes inside the system, wastewater, with mean concentrations (mg L⁻¹) of COD: 70, T-N: 12, T-P: 0.9, was introduced into the system at a loading rate of 1,000 L m⁻² d⁻¹. Treatment processes in the MSL system were different for the COD, P and N pollutants. Eighty percent of COD was removed in the 1st soil layer among the 6 layers, and the removal rate increased as water moved down and finally reached 90% in the last layer of the system. Phosphorus concentration was lower under the soil mixture layers than under the permeable layers, presumably because P was adsorbed mainly by soil and mixed iron particles. The P concentration in water gradually decreased in the lower layers of the system. The concentration of PO₄^3--P was generally lower in the aerated MSL system than in the non-aerated one. NH₄⁺-N was adsorbed and nitrified in the upper part of the system. The NO₃⁻-N concentration was lower in water under the soil mixture layers than under the permeable layers, indicating that denitrification mainly occurred in the soil mixture layers.     

Effects of Curtailment of Nitrogen Fertilizer on Biological Properties and Tea Leaf Yield in Acid Tea Field Soils

In Japan, the soils in tea fields are further acidified by heavy application of nitrogen fertilizer. In the long-term fertilizer application fields, available P, total carbon and nitrogen contents had the highest in the Multi-F plot, followed by the 98-D.F plot and the smallest in the 95-D.F plot. Soil enzymes activity had the highest the Multi-F plot. It was estimated that all of the enzyme activity were often corresponded with the results of the organic matter and the available P contents. The number of fungi under acidic condition had the close relation since change of these enzymes activity. It was evident that fungi were participated in carbon, nitrogen and phosphorus metabolism in acid tea field soils. It became clear that the number of fungi and enzymes activity decreased corresponding well with the available P and the organic matter contents depend on fertilization management. Moreover in case of qualities of green tea, generally difference between experimental plots and Multi-F plot were to be small. It was indicated that influence to the yields by curtailment of the amount of nitrogen fertilization was small.     

Detection of chromogranin A in the adrenal gland extracts of different animal species by an enzyme-linked immunosorbent assay using Thomsen-Friedenreich antigen-specific Amaranthus caudatus lectin

The reactivity of different lectins with crude chromogranin A (CgA) obtained from different animals, namely, cow, horse, dog, pig, and dolphin, was examined to identify lectin(s) that would be useful as coating reagent(s) in a sandwich enzyme-linked immunosorbent assay (ELISA). Of the different lectins studied, the Amaranthus caudatus lectin (ACA), which is specific for the Thomsen-Friedenreich (T)-antigen (Galβ1-3GalNAc), was found to react with the CgA from different animals by western blotting. Purified rabbit anti-bovine CgA antibody was also found to cross-react with the crude CgA preparations. On the basis of these findings, a sandwich ELISA was developed with ACA as the coating reagent and anti-bovine CgA antibody as the probing antibody. Using this method, concentration-dependent curves ranging from 0.003 μg/mL to 25 μg/mL and from 0.02 μg/mL to 25 μg/mL were obtained for bovine CgA and canine CgA, respectively. Similarly, concentration-dependent curves were obtained for the equine, swine, and dolphin crude CgA extracts. Thus, ACA is concluded to be a valuable reagent for CgA detection in crude extracts from different animal species, and for CgA isolation/purification.     

Persistent Contamination of Octopuses and Mussels with Lipophilic Shellfish Toxins during Spring Dinophysis Blooms in a Subtropical Estuary

This study investigates the occurrence of diarrhetic shellfish toxins (DSTs) and their producing phytoplankton species in southern Brazil, as well as the potential for toxin accumulation in co-occurring mussels (Perna perna) and octopuses (Octopus vulgaris). During the spring in 2012 and 2013, cells of Dinophysis acuminata complex were always present, sometimes at relatively high abundances (max. 1143 cells L⁻¹), likely the main source of okadaic acid (OA) in the plankton (max. 34 ng L⁻¹). Dinophysis caudata occurred at lower cell densities in 2013 when the lipophilic toxins pectenotoxin-2 (PTX-2) and PTX-2 seco acid were detected in plankton and mussel samples. Here, we report for the first time the accumulation of DSTs in octopuses, probably linked to the consumption of contaminated bivalves. Perna perna mussels were consistently contaminated with different DSTs (max. 42 µg kg⁻¹), and all octopuses analyzed (n = 5) accumulated OA in different organs/tissues: digestive glands (DGs) > arms > gills > kidneys > stomach + intestine. Additionally, similar concentrations of 7-O-palmytoyl OA and 7-O-palmytoly dinophysistoxin-1 (DTX-1) were frequently detected in the hepatopancreas of P. perna and DGs of O. vulgaris. Therefore, octopuses can be considered a potential vector of DSTs to both humans and top predators such as marine mammals.