Mediator-type biosensor for real-time wireless monitoring of blood glucose concentrations in fish

We developed a wireless mediator-type biosensor system to rapidly monitor blood glucose concentrations in free-swimming fish. Glucose oxidase was used to determine the glucose concentration, but fluctuations in the oxygen concentrations in the samples affected the measurements. In this work, we used a mediator, a synthetic electron acceptor, as a substitute in the oxidation–reduction reaction, thus ensuring that changes in the concentration of oxygen do not affect the measurements of glucose concentration performed by the sensor. We especially focused on the use of ferrocene as a mediator. The resulting enzyme sensor utilized a Pt–Ir wire (diameter 0.178 mm) as the working electrode and Ag/AgCl paste as the reference electrode. Chitosan was used to fix ferrocene and glucose oxidase to the working electrode. The characteristics of the sensor were tested, and it was found to be capable of measuring glucose independent of the oxygen concentration of the sample. Its reproducibility was also tested, and the results showed that the sensor was suitable for performing measurements in vivo. The sensor was then inserted into to eyeball interstitial sclera fluid in order to wirelessly monitor the glucose concentration in free-swimming fish. Measurements were taken for a total of 48 h. During these measurements, artificial stress was applied to the fish. The glucose concentrations of fish rise with their stress levels. The output current from the sensor gradually increased during the application of stress, which hinted that the stress was monitored by this system.     

基于ZigBee的水产养殖水环境无线监控系统设计

设计了一种基于ZigBee协议的水产养殖水环境无线监控系统,实现了对溶解氧、pH值、温度等多参数的采集、处理和显示,并通过无线网络实现了传感器检测节点和协调器节点之间数据快速、准确的传输,进而对多参数进行实时远程监测。该系统适用于工厂化水产养殖、水环境、智能温室等诸多领域。     

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.     

Use of an optical biosensor with a silicone-immobilized enzyme to determine plasma total cholesterol concentrations in fish

Decreased plasma total cholesterol concentrations are a useful indicator in fish of reduced resistance to bacterial infection. Taking advantage of the hydrophobic properties of silicone rubber, we developed an optical enzyme sensor system for measuring plasma cholesterol concentrations in fish. The system was constructed using an immobilized enzyme membrane, optical oxygen probe, flow system, and personal computer. The enzyme membrane was prepared from cholesterol esterase, cholesterol oxidase, and silicone rubber. The calibration curves were linear in the range of 55–85 mg dl⁻¹ for yellowtail and 20–90 mg dl⁻¹ for rainbow trout. Assays could be completed within 3 min, and the sensor response was stabilized by the hydrophobic properties of silicone rubber. Storage of immobilized enzyme membranes at 5°C permitted stable measurements for 58 days. The sensor system was applied to determine plasma total cholesterol concentrations in fish. Good correlations were obtained between results obtained using the sensor and results obtained using conventional methods (correlation coefficients: yellowtail 0.9751, rainbow trout 0.9947). Our method required much less time than conventional cholesterol assays and can be used economically for continuous determination of plasma total cholesterol in fish.     

基于Zigbee和GPRS的水质监测系统节能设计

针对无线化的水产养殖水质监测系统耗能大、电池寿命短的问题,设计了基于Zigbee和GPRS的节能型水质监测系统。通过采用低功耗器件,在电源与传感器、信号调理电路之间添加选通芯片ADG1414控制各模块分时分区工作,减少各模块的供电时间来降低硬件能耗;通过设置阈值对采集的数据进行判断,对阈值范围内的数据不发送,减少数据发送量,从而减少系统数据发送能耗。以CC2530为核心构建无线传感网络,将传感器采集到的温度、p H、溶氧等水质参数传输至监测中心,构建实时监测平台,并在此基础上建立数据管理系统,实现对水产养殖水质环境的实时监测。系统测试与实验结果表明,该系统节能效果显著,能有效延长无线水质监测系统电池的工作时间。     

Seasonal changes in vertical distribution patterns of cod (Gadus morhua) in the Bornholm Basin,central Baltic Sea

Hydroacoustic single fish detection and corresponding hydrographic measurements were used to study seasonal changes in vertical distribution of adult cod (Gadus morhua) in relation to ambient environmental conditions in the Bornholm Basin, central Baltic Sea. Sampling was conducted in April, June and August covering the years 2006–2009. Vertical distribution of individual fish was resolved from hydroacoustic single‐target detection in combination with a fish‐tracking algorithm and related to ambient hydrographic conditions. Based on a generalized linear effect model, both salinity and oxygen concentration were identified as key parameters affecting cod vertical distribution. Results also showed a clear seasonal effect with a more shallow distribution as the spawning season progressed and oxygen concentrations in the deep parts of the basin deteriorated. The upper limit of the distributional range was mostly constituted by the halocline and remained rather constant, whereas increasing oxygen depletion in the deep water layers lifted the lower boundary of the vertical distribution, leading to the observed upward shift in the overall distribution pattern. The results presented in this study highlight a significant shortcoming of the assessment survey design established for this species, as the observed shift in vertical distribution is not taken into account, thus introducing a potential bias into a data series used to tune the ICES standard stock assessment of this species.     

越冬池溶解氧含量的无线监测与控制系统初步设计

为确保鱼类安全越冬,避免因越冬池缺氧而大量死亡,设计了越冬池溶解氧含量无线监测和增氧机控制系统。该系统运行安全可靠,能够保证鱼类越冬生产的安全性,提高鱼类越冬成活率。     

基于分布式无线网络的水质监控系统设计

为提高对水产养殖水质监控的实时性和测量精度,设计了一种基于无线传感器网络的水产养殖水质参数监控系统。该系统由水质参数采集终端、分布式传感器网络、传输控制中心基站、远程在线监控系统组成。参数采集终端采集水质参数并传输到中心基站,再通过GPRS发送给远程在线监控中心,根据用户向监控中心输入的参数实现水温、pH、溶氧(DO)的调节。参数测量过程中引入数字滤波算法提高测量精度,使用经过改进粒子群优化算法(PSO)整定的PID控制器实现水质参数的调节。结果显示:测量精度达到要求,温度、pH和DO的测量误差分别为2.1%、1.3%和3.6%,系统对温度、pH和溶氧调节的最大误差分别为1.9%、2.6%和3.1%。整个系统工作稳定可靠。     

Estimating oxygen consumption of rainbow trout (Oncorhynchus mykiss) in a raceway: A Precision Fish Farming approach

The Precision Fish Farming (PFF) approach was applied to the estimation of fish oxygen consumption of rainbow trout in a raceway farm. A dynamic model, simulating the evolution of Dissolved Oxygen concentration, was identified: the daily oscillation of fish oxygen consumption rate was simulated by means of a sinusoidal function. The model was applied to the data set collected during a four-week field study, which was carried out in July 2019. Water temperature and Dissolved Oxygen concentration were measured with an hourly frequency in farm influent and effluent. Fish biomass was monitored on a daily basis by combining the data provided by a state-of-the art system for non-invasive estimation of fish weight distribution with mortality counting. The monitoring period was partitioned into two time-windows, as fish was not fed during the first two weeks. These windows were further partitioned into a calibration and validation set. Three model parameters, i.e. the average daily respiration rate, the amplitude of its daily oscillation, and its phase were estimated by fitting the model output to the time series of DO concentration in the effluent. The results of the calibration show that: 1) the daily average oxygen consumption rate is consistent with the literature; 2) the amplitude of the daily oscillation when fish is regularly fed is more than twice that estimated for fasting fish. The results of the validation suggest that the model could be used to implement a cost-effective automatic control of oxygen supply, based on the short-term prediction of oxygen demand.     

无线传感器网络技术在养殖业污染监测上的应用

综述了国际上无线传感器网络技术发展的特点和趋势,提出无线传感器网络技术在畜禽和水产养殖业污染监测上应用需要解决的2个技术问题:一是选择合理的IP无线传输方式;二是选择合理的系统结构,达到较高的性价比。设计出适合不同地域、不同生态环境的无线传感器网络的拓扑结构,实现在户外或野外环境下对畜禽和水产养殖业污染的监测。     

Acid–base balance in sea bass (Dicentrarchus labrax L.) in relation to water oxygen concentration

The influence of water oxygen concentration on the acid–base balance of sea bass was evaluated. Fish weighing 200–250 g were cultured under different dissolved oxygen concentrations of 64%, 97%, 150% and 250% saturation (92.7, 140.5, 217.5 and 362.7 mmHg respectively) under mild hypoxia, normoxia, mild hyperoxia and high hyperoxia conditions. The results showed that high hyperoxia and mild hypoxia conditions modified some blood parameters significantly when compared with fish held under the normoxia condition, while no differences were shown with respect to the acid–base balance of fish cultured under normoxia and mild hyperoxia conditions. This testifies that the mild hyperoxia condition does not produce physiological disturbances in the acid–base status of sea bass and it could be considered a favourable condition in sea bass land‐based farming, mainly in comparison with the mild hypoxia condition, responsible for other physiological problems.     

Effective monitoring of freshwater fish

Freshwater ecosystems constitute only a small fraction of the planet's water resources, yet support much of its diversity, with freshwater fish accounting for more species than birds, mammals, amphibians or reptiles. Fresh waters are, however, particularly vulnerable to anthropogenic impacts, including habitat loss, climate and land use change, pollution and biological invasions. This environmental degradation, combined with unprecedented rates of biodiversity change, highlights the importance of robust and replicable programmes to monitor freshwater fish. Such monitoring programmes can have diverse aims, including confirming the presence of a single species (e.g., early detection of alien species), tracking changes in the abundance of threatened species, or documenting long‐term temporal changes in entire communities. Irrespective of their motivation, monitoring programmes are only fit for purpose if they have clearly articulated aims and collect data that can meet those aims. This review, therefore, highlights the importance of identifying the key aims in monitoring programmes and outlines the different methods of sampling freshwater fish that can be used to meet these aims. We emphasize that investigators must address issues around sampling design, statistical power, species’ detectability, taxonomy and ethics in their monitoring programmes. Additionally, programmes must ensure that high‐quality monitoring data are properly curated and deposited in repositories that will endure. Through fostering improved practice in freshwater fish monitoring, this review aims to help programmes improve understanding of the processes that shape the Earth's freshwater ecosystems and help protect these systems in face of rapid environmental change.     

Computer Analysis of Fish Populations in Fluctuating Streams

POPULATION ESTIMATES, a computer program designed to analyse fish populations in fluctuating streams, is described. Its special feature is to calculate population density (number and g m^?2) as a dynamic function of variable stream area. Other output includes estimated population size, mean length, mean weight and mean condition factor. Simulated flow changes of 12.5–400% changed stream surface area 6–37% for straight channels, 11–80% for meandering channels and 18–182% for braided channels. Fish population density, however, may not change inversely proportional to stream size. The main use of POPULATION ESTIMATES is to evaluate the effects of fluctuating flows on fish populations.     

换水换气式充氧密封鱼苗运输装置及其应用的研究

换水换气式充氧密封鱼苗运输装置是直接在塑料袋运输活鱼基础上改装而成，它能在不松开塑料袋的条件下，轻松快速地进行换水换气。通过以鳙鱼苗为试验对象，选用换水量、换气量、换水间隔期和换气间隔期四因素模拟运输进行了该装置的九组正交试验，得出了四因素最经济可行的组合是换水量为60％，换水间隔期为16小时，换气量为50％，换气间隔期为8小时。正交试验中存活时间最长为55小时（空白试验为26小时），换水量是最主要的影响因素，只换水比只换气更利于延长活鱼运输时间。     

基于Bytetrack的多目标跟踪算法在斑马鱼毒性行为识别中的应用

利用计算机视觉技术识别斑马鱼(Danio rerio)在不同污染物暴露下的行为变化是水质毒性评价的常用方法之一,但传统方法存在效率低、面对遮挡和复杂环境时性能差等缺陷。针对这些问题,本研究使用基于Bytetrack的多目标跟踪算法追踪斑马鱼在4种污染物(Zn、Pb、Cr和苯酚)暴露2 h后的行为变化,对斑马鱼在4种浓度梯度中的平均速度、最大速度、最低速度、平均碰撞次数和行为轨迹等指标进行分析。结果显示,算法的追踪精度、漏检率和检测时间(每300帧)分别能达到90.26%、16.33%和0.19 min,检测时间和精度相比于传统目标检测方法有较大提升。同时,根据污染物不同,该方法能准确识别特定污染物环境中斑马鱼相应的运动状态及轨迹变化,可实现精确识别和实时响应,在鱼类毒性行为识别领域具有重要参考意义。     

Enzymatic Hydrolysis of Alaska Pollock Proteins Based on Kinetics Model and Lysine Biosensor–Neural Network Model

Controlled proteolysis is important for target hydrolysates. The hydrolysis kinetics of pollock bone protein (PBP) was obtained by mathematic deduction and experimental analysis. The equation of degree of hydrolysis was as follows: DH = 2.801 ln|1 + (0.06044E₀/S₀?0.1005)t|, which could predict the hydrolysis process of PBP when hydrolysis condition was strictly controlled. However, the hydrolysis reaction was affected by several factors, and the predicted value may be against the experimental result. Therefore, online monitoring was necessary for obtaining the target peptides. Biosensor and artificial neural network (ANN) were employed for monitoring hydrolysis process online. Free lysine level was chosen as the monitoring factor determined by immobilized lysine oxidase electrodes. Based on the lysine biosensor and ANN, the hydrolysis monitoring model LYS-ANN was built, including 3 input layer nodes (E₀, S₀, and LYS), 1 output layer node (DH), and 11 hidden layer nodes. R² value between sample value and simulation value was 0.9964. Simulation error was in the range of 0–4.56%, and the average relative error was 0.94%. The verification tests of PBP hydrolysis showed that the LYS-ANN model could forecast hydrolysis process successfully with high efficiency and accuracy even if the hydrolysis conditions varied.     

基于FPGA的水产养殖环境无线测控模块设计

针对我国水产养殖无线远程信息测控多以成熟单片机为核心控制主件,不易满足行业特殊接口需求以及缺乏独立核心控制器的现状,项目以标准化及芯片自主化为最终目的,提出基于现场可编程门阵列(FPGA)为核心的实现方案,尝试性设计了一种较为通用的、可实现AD转换、数字接口、控制输出和驱动以及养殖现场数据存储和远距离数字通讯的测控模块。通过对水产养殖领域环境信息的无线测控模块各主要环节的研究设计,以模块到系统的FPGA原型功能验证方式,实现了现场模块对水产养殖的温度、溶氧信息的远距离测量和控制。系统测试和板级实验结果表明,该设计可以满足低成本、接口可扩展及标准化核心控制器的水环境测控模块需求。     

The effect of ploidy and temporal changes in the biochemical profile of gibel carp (Carassius gibelio): a cyprinid fish species with dual reproductive strategies

The basic biochemical profile of gibel carp (Carassius gibelio), diploid–triploid cyprinid species complex with dual reproduction modes, was investigated. The aim of the study was to test for the effect of fish group (which combines the effects of ploidy and sex) and temporal changes on indicators of the biochemical profile of blood in gibel carp. We showed that the basic biochemical profile of blood is significantly influenced by season and fish group. Triploid gynogenetic females had a better condition measured by total protein concentration than sexual diploids. The higher concentrations of triacylglycerols and cholesterol in triploid females when compared to diploids indicate a higher metabolic rate and higher energy intake during the whole year, which may represent a disadvantage for triploids. Differences in the concentrations of blood biochemical analytes between triploid females and diploids suggest a potential difference in the regulation of a wide range of physiological and biochemical processes in which these analytes participate. The biochemical profiles of two forms of C. gibelio could at least partially explain the changes of character of its naturally occurring populations.     

Thermal stress altered growth performance and metabolism and induced anaemia and liver disorder in Labeo rohita

Sudden fish deaths occurred during summer peaks in India and expected to be more in future. This study was conducted to explain the biochemical responses of Labeo rohita under extreme thermal condition (treated: 37–38°C against controlled: 28–30°C). Exposure of 14 days resulted in 30% of fish mortality. Glucose concentration was maintained in serum for both treated and controlled groups. However, triglycerides, protein, globulin, calcium, cholesterol and haemoglobin were declined and enzymes (glutamate‐pyruvate transaminase, GPT and glutamate‐oxalate transaminase, GOT) were elevated in serum in the treated group. Indicators of overall metabolism (biomolecules: glucose, triglycerides, protein, cholesterol; enzymes: GPT, GOT; and RNA: DNA) were diminished in liver, whereas these were less affected in muscle. Higher GPT, GOT and lower albumin in serum confirmed disturbance in hepatic panel. Simultaneously compromised growth (lower SGR, DNA content in liver) was observed in treated fish due to impaired metabolism. Extreme thermal stress induced by such catastrophic warmer water exposure caused liver problems and anaemia which ultimately caused fish death.     

全双工射频识别技术的过鱼效果监测与分析

全双工射频识别系统(RFID)相较于半双工具有操作简便、数据传输方便等优势,国外广泛应用于鱼道过鱼效果评价。针对而国内相关研究较少的现状,以植有PIT标签的仿野生齐口裂腹鱼(Schizothorax prenanti)识别与否作为评判系统监测效率高低的依据,定量分析鱼体入射角度、鱼体入射范围、鱼体长大小和游泳速度对全双工射频识别系统监测效率的影响,以接收到监测数据作为因变量,构建Logistic回归模型,利用AIC模型准则筛选出最优的Logistic回归模型。结果表明,游泳速度(P=0.001)和鱼体长大小(P=0.03)与系统成功监测概率呈负相关。鱼体入射角度(0°～90°)对系统监测效率的影响不显著(P0.05);鱼体入射范围0～10 cm的监测效率显著高于入射20～30 cm和30～40 cm(P0.05)。为进一步提高鱼道监测效率,建议在开展实际鱼道工程监测前,应掌握鱼类基础参数(如体长)和行为(如游泳速度)数据,合理布置天线位置,将有助于为监测评估后的鱼道优化设计提供技术支撑和数据参考。