Wireless monitoring of blood glucose levels in flatfish with a needle biosensor

A wireless biosensor system was developed for the continuous measurement of blood glucose levels in flatfish. The biosensor was implanted in the interstitial fluid under the scleral surface of the eyeball (EISF) to investigate the relationship between EISF and blood glucose levels. EISF glucose levels were found to be correlated with those in the blood and to be approximately the same as blood glucose levels in the range of 7–25 mg dl⁻¹. A needle-type biosensor was prepared for the continuous EISF glucose monitoring in flatfish. A working electrode was constructed using platinum iridium wire, and glucose oxidase was immobilized to the electrode. The biosensor was inserted into the EISF of flatfish for sensor implantation. A 650-mV potential (vs. Ag/AgCl) was applied by a wireless potentiostat to the working electrode for the amperometric glucose measurement. We investigated whether glucose in the EISF can be determined in vivo. The estimated glucose levels using a one-point calibration method were correlated with actual blood glucose levels. In conclusion, using a wireless biosensor system, we were able to monitor blood glucose levels in flatfish under free-swimming conditions for 16 h.     

Development of biosensor for measuring oxidative stress of fish

Fisheries Science - To elucidate the dynamics of oxidative stress in fish, it is necessary to know the concentration of superoxide anions as a precursor to various reactive oxygen species in the...     

Development of a label-free immunosensor system for detecting oocyte maturation-inducing hormone in fish

A label-free immunosensor for detecting the oocyte maturation-inducing hormone 17, 20β-dihydroxy-4-pregnen-3-one (DHP), was developed. A principle of the sensor system was based on differences in electrochemical activity changed by an immunoreaction in the absence and presence of DHP. For preparation of the immunosensor, anti-DHP IgG was immobilized on an Au working electrode modified with a self-assembled monolayer of 3-mercaptopropionic acid. The sensor was immersed into a sample solution containing DHP. DHP was determined by cyclic voltammetry. The immunosensor showed a specific response to DHP, and the oxidation peak current linearly decreased in the range of 7.8–500 pg ml⁻¹ with a correlation coefficient of 0.996. The sensor system was applied to determine the DHP levels in plasma of goldfish and was compared with the DHP levels of the same samples determined using an ELISA as the conventional method. Good correlation was obtained between values determined using both methods in the range of 0.1–7.7 ng ml⁻¹ (correlation coefficient 0.876). These findings suggest that the proposed label-free immunosensor can be used to analyze DHP levels in fish plasma samples.     

Physiological characteristics of blue sac disease in chum salmon alevins

Fisheries Science - Blue sac disease (BSD) in salmonids results in the abnormal accumulation of ascites fluid and can often lead to mass mortality. However, the symptoms and causes of BSD remain...     

The effects of organic carbon on acid rain in a temperate forest in Japan

From recent studies, we noticed that stemflow had an acidity that differed from that of precipitation or throughfall. Organic substances, supplied from the tree surface, would be one of the factors that modifies the acidity of rain. The objectives of this study were to determine the DOC concentration and to clarify the influence of dissolved organic carbon (DOC) on acidity in precipitation, throughfall and stemflow. Throughfall and stemflow were measured in sugi [Cryptomeria japonica D. Don], hinoki [Chamaecyparis obutusa Endl.] and kojii [Castanopsis cuspidata (Thumb.) Schottky.] stands. All samples were analyzed for their pH, electric conductivity (EC), major inorganic anions and cations and DOC concentration.The annual average of DOC was highest in stemflow, and that of throughfall and precipitation were one-third and one-tenth of stemflow, respectively. The averages of DOC in stemflow in two coniferous, sugi and hinoki stands, were higher than that of broadleaved kojii stand. DOC concentration was low in summer and high in winter in all stands. In Stemflow, pH and DOC were negatively correlated, while EC and DOC in stemflow were positively correlated in all stands. However in throughfall, there was no evident relationship between pH, DOC and EC. This relationship was not explained by the cause of organic acid.     

Carbon nanotube enhanced label-free immunosensor for amperometric determination of oocyte maturation-inducing hormone in fish

Maintaining high-quality fish eggs stably and efficiently is important for aquaculture. We developed a label-free immunosensor system for measuring 17,20β-dihydroxy-4-pregnen-3-one (DHP). DHP is suddenly secreted before ovulation as a maturation-inducing hormone in fish, and therefore, DHP levels are an indicator for predicting ovulation. The method is based on immunologic reactions and amperometric measurement using cyclic voltammetry (CV). For biomolecular immobilization on the surface of sensing electrode, Au electrode, we used self-assembled monolayers of thiol-containing compounds to fix anti-DHP immunoglobulin. In addition, we used a single-walled carbon nanotube to improve sensitivity. Using this electrode, we were able to determine the CV signal change caused by the antigen–antibody complex. The proposed immunosensor system showed a linear correlation (correlation coefficient: 0.9827) between the anodic peak current of the CV and the DHP level in range from 15.6 to 50,000 pg ml^?1. The sensor system was then applied to monitor DHP of goldfish (Carassius auratus). Blood plasma of fish was collected every 3 h after administering a DHP inducer. In the measurement, the anodic peak current of the CV showed distinct changes depending on DHP levels in the blood plasma. A good relationship was observed between DHP levels determined by our proposed system and the conventional method (correlation coefficient: 0.9351).     

An evaluation of the contribution of hatchery stocking on population density and biomass: a lesson from masu salmon juveniles within a Japanese river system

Stocking is an important management tool for enhancing fisheries resources, but its actual contribution to fisheries resources is controversial, taking into consideration both the positive and negative effects. This study compared density and biomass of hatchery (otolith thermal marked) and wild masu salmon parr between stocked and unstocked rivers to evaluate the contribution of stocking with hatchery‐reared fish. Density and biomass of all fish did not differ between stocked and unstocked rivers. Moreover, density and biomass of wild fish in the stocked rivers were lower than those of the unstocked rivers. Density and biomass of hatchery fish in a non‐natural reproducing river were similar with those of all fish in natural reproducing rivers. These results indicate that hatchery stocking does not have positive effects on population density or biomass but replaces wild fish with hatchery fish and that non‐natural reproducing areas are more suitable as stocking sites.     

Development of a label-free immunosensor system for detecting plasma cortisol levels in fish

Fishes display a wide variation in their physiological responses to stress, which is clearly evident in the plasma corticosteroid changes, chiefly cortisol levels in fish. In the present study, we describe a novel label-free immunosensor for detecting plasma cortisol levels. The method is based on immunologic reactions and amperometric measurement using cyclic voltammetry. For the immobilization of the antibody on the surface of sensing electrode, we used a self-assembled monolayer of thiol-containing compounds. Using this electrode, we detect the CV signal change caused by the generation of antigen–antibody complex. The immunosensor showed a response to cortisol levels, and the anodic peak value linearly decreased with a correlation coefficient of 0.990 in diluted plasma. The specificity of the label-free immunosensor system was investigated using other steroid hormones, such as 17α, 20β-dihydroxy-4-pregnen-3-one, progesterone, estriol, estradiol, and testosterone. The specific detection of cortisol was suggested by a minimal change from ?0.32 to 0.51 μA in the anodic peak value of the other steroid hormones. The sensor system was used to determine the plasma cortisol levels in Nile tilapia (Oreochromis niloticus), and the results were compared with those of the same samples determined using the conventional method (ELISA). A good correlation was obtained between values determined using both methods (correlation coefficient 0.993). These findings suggest that the proposed label-free immunosensor could be useful for rapid and convenient analysis of cortisol levels in fish plasma samples.     

High level expression of C-terminal truncated recombinant chicken interferon-gamma in baculovirus vector system

Chicken interferon-gamma (ChIFN-gamma) was expressed by baculovirus in a C-terminal truncated form, namely ChIFN-gammaT, to accelerate the secretion of the expressed protein. It is also expressed as ChIFN-gammaT bearing poly His tag, ChIFN-gammaTHis, for easy purification. The expressed proteins were detected by SDS-PAGE analysis with Coomassie brilliant blue staining. The purified ChIFN-gammaTHis with nickel chelated column showed anti-viral activity in vitro and stimulation of the secretion of nitrogen intermediates such as nitric oxide in chicken peripheral blood mononuclear cells. Antiserum against ChIFN-gammaTHis recognized the 15 kDa, 16 kDa, and 32 kDa bands that seemed to be an unglycosylated monomer, a glycosylated monomer, and a homodimer of ChIFN-gammaTHis in the culture supernatant, respectively. The anti-serum also recognized around 14 kDa and 28 kDa bands in the sera of chickens or concanavalin A stimulated spleen cell culture supernatants that seemed to be monomeric and dimeric forms of a natural ChIFN-gamma, respectively.     

Seasonal fluctuation of groundwater in an evergreen forest, central Cambodia: experiments and two-dimensional numerical analysis

This study of a water cycle was conducted in an evergreen forest located in the Mekong River Basin in central Cambodia. At the observation site, we measured the dynamics of the spatial distribution of groundwater levels. The groundwater movement was analyzed two-dimensionally using boundary conditions and parameters that had been observed in the field. The climate in the research area is dominated by two seasons, which occur annually: a rainy and a dry season. The groundwater levels are generally high during the rainy season and low during the dry season. Groundwater levels were measured along a stream, which flowed through the study site. The streambed was visible at the head of the stream in January. At the next downriver well point, the streambed appeared in March. Finally, it became visible at all well points in April, meaning that surface runoff had disappeared temporarily and instead flowed underground during the ensuing dry period. Groundwater levels of the studied lateral flow perpendicular to the stream that seeped and infiltrated into the stream were 1.2–2.5 m deep (in April), which was the lowest level recorded for the year. During that period, the depth of the groundwater of the studied lateral flow fell by as much as 56 mm per month. In addition, the lateral flow groundwater infiltrated into groundwater of the stream during that period. The groundwater level fluctuation was estimated based on a two-dimensional analysis of lateral flow perpendicular to the stream using a numerical simulation model with soil physical parameters and observed boundary conditions. The observations of ground water fluctuations were well reproduced. Deep seepage of groundwater was estimated using a uniform boundary condition that allowed efflux through the bottom, estimated as being approximately 30 mm per year. The simulated deep seepage rate was considered plausible considering other hydrological components such as soil water storage fluctuation.