The ISF in fish is found behind the eyeball and can be accessed b

The ISF in fish is found behind the eyeball and can be accessed by gently pushing the eyeball to the side near the snout. We confirmed that glucose levels selleck chemicals llc in the ISF are highly correlated with those in blood based on 112 samples of fish eyeball ISF (y = 2.2996 + 0.9438x, R = 0.960) [8]. We named this ISF ��eyeball interstitial sclera fluid�� (EISF). We then developed a fish body-implantable needle-type glucose biosensor connected to a wireless monitoring system [9]. Implantation of the biosensor in the EISF allowed for real-time monitoring of blood glucose levels in fish. The needle-type glucose biosensor we developed is a dipole biosensor comprising a platinum iridium wire as the working electrode and Ag/AgCl as the counter electrode [8�C11].
To limit the effect of impurities in the EISF, a 5% Nafion solution was applied to the working electrode and glucose oxidase (GOx) was immobilized onto the electrode by forming an enzyme layer on top of the Nafion layer. The sensor output current in glucose standard solutions had a very strong correlation (R = 0.9983) with glucose concentrations within the range of 0.18 to 144 mg?dL?1 [9]. The blood glucose level of the fresh water fish (tilapia) is reported to be ~50 to 100 mg?dL?1 [12]. Therefore, this sensor is applicable for measuring glucose levels in fish. By connecting the sensor to a wireless, radio wave-based monitoring system and, we were able to continuously measure blood glucose levels for 72 h [9].In 1996, Yoon et al. developed a stick-type biosensor for measuring l-lactic acid concentrations [13].
A method for measuring blood l-lactic acid concentrations in fish, however, has not been reported to date.Based on our understanding of fish stress responses during breeding, real-time monitoring of blood l-lactic acid concentration and blood glucose levels in fish will contribute to maintaining disease-free fish aquaculture. The purpose of this study was to develop a method for real-time monitoring of l-lactic acid concentrations in the fish blood using a biosensor. We first investigated the relationship between EISF and blood l-lactate acid levels in fish. We then developed a needle-type enzyme sensor, evaluated the performance of this sensor, and examined the possibility of measuring l-lactic Dacomitinib acid concentrations in fish blood using a wireless monitoring method.2.?Experimental Section2.1.
ReagentsLactic acid oxidase (LOX) from Pediococcus sp.; E.C. 1.13.12.4, 100 U/g was selleck purchased from Sigma-Aldrich (St. Louis, MO, USA). Bovine serum albumin (BSA), 5% Nafion dispersion solution, 2-phenoxy ethanol, and heparin sodium (10,000 U/g) were purchased from Wako Pure Chemical Industries (Tokyo, Japan). Glutaraldehyde solution (25%) was purchased Tokyo Chemical Industries (Tokyo, Japan). A lactic acid standard solution was prepared by dissolving lactic acid in 0.1 M phosphate-buffered saline (PBS).2.2.

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