A biotelemetry system recording fish activity

A biotelemetry system is described for obtaining, transmitting and recording the electromyograms (EMGs) produced in muscle activity of free-swimming fish as quantitative indicators of overall fish activity. The radiotransmitters used come in the form of cylindrical packages having two sensing electrodes, all fully implantable in the fish body cavity. EMGs are transmitted as radio pulses with the intensity of muscular activity determining the intervals between pulses. The packages also contain temperature sensors and fish temperatures are transmitted with every 32nd pulse. Transmitted EMG pulses are detected, 'measured' and stored by a single portable receiver (Model SRX_400, Lotek). Data can be subsequently transferred to a computer (which can also be portable) for storage, processing and statistical analysis. Transmitter battery life can be in excess of 7 months, permitting laboratory or field studies of long duration. Transmitter package implantation surgery requires a mid-ventral incision and internal securing of transmitter and sensing electrodes. Surgical silk, cyanoacrylate tissue adhesives, and polydioxanone (PD), a synthetic absorbable suture, were all tried as means of incision closure. The most effective was PD alone. Trials of the system consisted of forced swims by transmitter-equipped rainbow trout Oncorhynchus mykiss Walbaum. The data obtained provided an inverse linear relation between forced swim speed and EMG pulse interval. Trials were conducted at intervals over periods up to 2 months. Fish showed neither distress, nor difficulty in swimming up to maximum speeds of 60 cm s ⁻¹ (fish lengths 41.0, 44.4 cm).     

An integrated mini biosensor system for continuous water toxicity monitoring

An integrated water toxicity monitoring system that uses recombinant bioluminescent bacteria was successfully developed for the continuous monitoring and classification of toxicities present in water. This system consists of four channels arranged horizontally inside of a cylinder, with each channel having two small bioreactors that are vertically connected to each other to maintain a separation of the culture reactor from test reactor. This system is easily handled and installed, making its application in the field a potential reality. As well, it performed stably and continuously due to the vertical separation of the culture reactor from the test reactor and a long term operation was also performed because of its small working volume, i.e., only 1 ml for the 1st bioreactor and 2 ml for the 2nd. During an operation with four strains, i.e., EBHJ2, DP1, DK1 and DPD2794, which are responsive to superoxide damage (EBHJ2 and DP1), hydrogen peroxide (DK1), and DNA damage (DPD2794), the O.D. and bioluminescence of the bacterial cultures inside the system were constant when no chemical was injected. However, with the addition of paraquat, hydrogen peroxide or mitomycin C, the bioluminescent responses of the strains were found to be dose-dependent to different concentrations of these chemicals.     

Monitoring fish behaviour with a remote, combined acoustic/radio biotelemetry system

Biotelemetry is a powerful instrument for monitoring aquatic species in their natural environment. Using telemetry, animals can be monitored from a passive perspective, without the biases associated with conventional handling and sampling techniques. To monitor aquatic species in remote environments, with vast stretches of water, and in situations requiring both acoustic and radio transmissions (e.g. for diadromous fish), advances in telemetry are necessary. In this paper, a field-proven telemetry system based on a radio receiver and incorporating combined acoustic and radio smart transmitters, wireless hydrophones and two-way satellite communications is described. The system was first deployed in Bay d'Espoir, Newfoundland, Canada, in 1998. The purpose of this deployment was to determine whether aquaculture triploid steelhead trout ( Oncorhynchus mykiss Walbaum) (1.5–2.0 kg), experimentally released in the vicinity of a commercial aquaculture site, remained at the site (site fidelity) or dispersed. Two sets of fish releases, summer and winter, were performed to determine seasonal effects on the movement of aquaculture triploid steelhead trout in the wild. The results suggested strong site fidelity among steelhead trout when released during the growing season (summer). However, less fidelity was displayed for the winter released steelhead.     

Biosensor system for continuous monitoring of organophosphate aerosols

An enzyme-based monitoring system provides the basis for continuous sampling of organophosphate contamination in air. The enzymes butyrylcholinesterase (BuChE) and organophosphate hydrolase (OPH) are stabilized by encapsulation in biomimetic silica nanoparticles, entrained within a packed bed column. The resulting immobilized enzyme reactors (IMERs) were integrated with an impinger-based aerosol sampling system for collection of chemical contaminants in air. The sampling system was operated continuously and organophosphate detection was performed in real-time by single wavelength analysis of enzyme hydrolysis products. The resulting sensor system detects organophosphates based on either enzyme inhibition (of BuChE) or substrate hydrolysis (by OPH). The detection limits of the IMERs for specific organophosphates are presented and discussed. The system proved suitable for detection of a range of organophosphates including paraoxon, demeton-S and malathion.     

Enhancing the sensitivity of a two-stage continuous toxicity monitoring system through the manipulation of the dilution rate.

Optimization of the dilution rates has been studied to provide an enhanced sensitivity to toxicity by several recombinant bioluminescent Escherichia coli strains, TV1061 (grpE::luxCDABE), DPD2794 (recA::luxCDABE) and DPD2540 (fabA::luxCDABE), in the two-stage continuous toxicity monitoring system. It was found that the sensitivity of both TV1061 and DPD2794 to a pulse injection of phenol and mitomycin C increased with a decrease in the dilution rate. The sensitivity, however, for all the strains to step injections of the toxic chemicals was found to increase with an increase in the dilution rate up to a certain dilution rate and then decreased, mainly due to the rapid washing out of the injected chemicals. The response kinetics of the strains were explained by evaluating the mode of action of the recombinant bioluminescent bacteria to toxicity with the dilution rate, the operating parameter of minibioreactors under consideration in this study.     

A two-stage minibioreactor system for continuous toxicity monitoring.

A two-stage minibioreactor system was successfully developed for continuous toxicity monitoring. This system consists of two minibioreactors in series. Recombinant Escherichia coli DPD2794 containing a RecA::luxCDABE fusion as a model strain was utilized to monitor environmental insults to DNA, with mitomycin C as a model toxicant. Pulse type exposures were used to evaluate the system's reproducibility and reliability. Step inputs of mitomycin C have been adopted to show the system's stability. The system's ability to monitor the possible upsets or accidental discharges of toxic chemicals was also evaluated with these step insults. All the data demonstrated that this two-stage minibioreactor system using recombinant bacteria containing stress promoters fused with lux genes is quite appropriate for continuous toxicity monitoring. Long-term operation and minimized media-usage have been investigated. Thus application to many different areas, including an early warning system of wastewater biotreatment plant upsets and the monitoring and tracking of accidental spills, discharges or failures in plant operation are plausible.     

A novel continuous subcutaneous lactate monitoring system

A novel continuous lactate monitoring system has been developed modifying the GlucoDay portable medical device (A. Menarini Diagnostics), already present in the European market, and used to continuously measure glucose levels. Lactate oxidase based biosensors have been developed immobilising the enzyme on nylon net and placing it on a Pt electrode. The biosensor was connected to the portable device provided with a micro-pump and coupled to a microdialysis system. It is capable to record subcutaneous lactate every 3 min. In vitro analytical results confirmed that the sensors respond linearly in the interval of concentration between 0.1 and 10 mmol/L, covering the whole physiological range. During prolonged monitoring periods, the response of the biosensors remained stable, showing a limited drift of 8%, within 60 h. Stability tests are still on route. However, preliminary results have shown a shelf life of about 10 months. In vivo experiments performed on healthy rabbits have demonstrated the good accuracy and reproducibility of the system. A correlation coefficient equal to 0.9547 (N=80) was found, which represents a good correlation between the GlucoDay and the laboratory reference analyser. A 16 h in vivo monitoring on a healthy volunteer has been also performed.     

The fate of dead fish tagged with biotelemetry transmitters in an urban stream

Mortality is a key factor in understanding the population dynamics of fish. For studies using biotelemetry, missing individuals pose a challenge since the ultimate fate of both the animal and the tag are unknown. In this study, we document three releases of carcasses in a simulated small-scale summer fish kill in a small urban stream using juvenile white sucker (Catostomus commersonii). Passive integrated transponder (PIT) and radio tags were affixed to carcasses that were tracked to determine the fate of both the dead fish and the tags. Mean daily dispersal distances ranged from 0.0 to 7.6 m day^?1 and included downstream and lateral dispersal. Of the 44 radio-tagged carcasses, 26 tags (and presumably carcasses) were consumed by scavengers; the majority were consumed by snapping turtles (Chelydra serpentina) with fewer carcasses scavenged by great blue heron (Ardea herodias), raccoon (Procyon lotor) and muskrat (Ondatra zibethicus). We also contrasted the decomposition rates of in-stream carcasses with those experimentally placed on the riverbank and found that while there was no significant difference in the time to carcass evisceration, the rate of decomposition was more rapid in-stream compared to on-shore. Radio tag loss during the decomposition study was moderate (one of three lost); PIT tag loss occurred when carcasses became eviscerated, typically by invertebrates. By examining the role of scavengers, dispersal and decomposition, it is possible to understand the fate of dead fish, the fate of tags and role of mortality in tagging experiments and the connections between stream and riparian habitats and organisms. This information will help inform the interpretation of potential mortalities in fish tracking studies and improve fish kill investigations.     

Wireless enzyme sensor system for real-time monitoring of blood glucose levels in fish

Periodic checks of fish health and the rapid detection of abnormalities are thus necessary at fish farms. Several studies indicate that blood glucose levels closely correlate to stress levels in fish and represent the state of respiratory or nutritional disturbance. We prepared a wireless enzyme sensor system to determine blood glucose levels in fish. It can be rapidly and conveniently monitored using the newly developed needle-type enzyme sensor, consisting of a Pt-Ir wire, Ag/AgCl paste, and glucose oxidase. To prevent the effects of interfering anionic species, such as uric acid and ascorbic acid, on the sensor response, the Pt-Ir electrode was coated with Nafion, and then glucose oxidase was immobilized on the coated electrode. The calibration curve of the glucose concentration was linear, from 0.18 to 144mg/dl, and the detection limit was 0.18mg/dl. The sensor was used to wirelessly monitor fish glucose levels. The sensor-calibrated glucose levels and actual blood glucose levels were in excellent agreement. The fluid of the inner sclera of the fish eyeball (EISF) was a suitable site for sensor implantation to obtain glucose sample. There was a close correlation between glucose concentrations in the EISF and those in the blood. Glucose concentrations in fish blood could be monitored in free-swimming fish in an aquarium for 3 days.     

An infrared system for monitoring Drosophila motility during microgravity

Presently, the precise mechanisms of the aging process are unknown. Examination and comprehension of the aging process in other species could lead to significant advances in the understanding of human aging. Drosophila melanogaster (fruit fly), commonly used for aging studies, is a widely studied organism in terms of behavior, development, and genetics. Previous microgravity experiments have shown a significant decrease in the life span of young male Drosophila after microgravity exposure. This decrease in lifespan may be related to locomotor activity, a convenient measure of overall physiological performance. This study describes the design and performance of a Drosophila Infrared Motility Monitoring System (DIMMS). The DIMMS uses a unique design of two infrared (IR) beams per fly to measure the locomotor activity of 240 flies. Locomotor activity is measured in terms of number of IR crossings per unit time, instantaneous velocity, and continuous velocity. Ground-based results using the DIMMS equipment agree well with previous values for Drosophila locomotor velocity. DIMMS is an improvement over equipment previously used due to its ability to continuously monitor locomotor activity throughout short-duration microgravity exposure. DIMMS is also lightweight, compact, and power efficient. DIMMS has been flight tested onboard NASA's KC-135 reduced gravity research aircraft and a Nike-Orion sounding rocket.     

A multichannel perifusion system for the continuous monitoring of glycogenolysis and gluconeogenesis in isolated rat hepatocytes

A multichannel perifusion system for isolated rat hepatocytes entrapped in a Sephadex matrix is described and criteria for the choice of matrices are discussed. This system overcomes the usual problem of clogged filters and impaired flow rates encountered in suspension perifusion systems, and is assembled from standard widely available components. Gluconeogenic capability and mitochondrial respiratory control ratios were unaltered. Decreases in trypan blue viability index and respiration rate were small when compared with flask-incubated hepatocytes. The endogenous rate of glycogenolysis was slightly higher in perifused hepatocytes but hormone response, as judged by glucagon stimulation of glycogenolysis, was unimpaired. The potential of this system is indicated by experiments monitoring glycogenolysis and gluconeogenesis in recycling and non-recycling modes.     

New detection system for toxic agents based on continuous spectroscopic monitoring of living cells

In this study, we show the feasibility of a new type of cell based biosensor which uses spectroscopic in situ real time detection of biochemical changes in living cells exposed to toxic chemical agents. We used a high power 785 nm laser to measure the time dependent changes in the Raman spectrum of individual living human lung cells (A549 cell line) treated with a toxic agent (Triton X-100, 250 microM solution). Individual cells were monitored by Raman spectroscopy over a total time span of 420 min, with 30 min sampling intervals. During this period of time, the A549 cells were maintained in a purpose designed temperature controlled cell chamber, which allowed the cells to be maintained in physiological conditions. The time dependent changes in the Raman spectra were correlated with the sequences of events that occur during cell death. The molecular mechanisms involved in cell death are indicated by the decrease in the magnitude of Raman peaks corresponding to proteins (1322, 1342 and 1005 cm(-1)) and DNA (decrease by 80-90% in the 786 cm(-1) phosphodiester bonds C'5-O-P-O-C'3). To support these conclusions, viability tests and Western blotting analysis of PARP protein were carried out. This technique could overcome the limitations of other detection systems available, since the specific time dependent biochemical changes in the living cells can be used for the identification and quantification of a large range of toxic agents. This technique could also be used with cellular microarrays for high throughput in vitro toxicological testing of pharmaceuticals and in situ monitoring of the growth of engineered tissues.     

A fast method for monitoring the colonization rate of lactobacilli in a meat model system

A random amplified polymorphic DNA (RAPD) assay coupled to a fast and reproducible cell lysis method from Lactobacillus colonies were developed to type lactobacilli of different strains and species, with the aim of precisely enumerating each of the different Lactobacillus strains inoculated in a nutrient-rich environment, such as sausage meat batter. Colonization assays were carried out in an aseptic meat fermentation system for up to 14 d and the inoculated strains were challenged with mixtures of wild lactobacilli. The proportion of inoculated strains remaining at different times was compared with the total number of lactobacilli grown on MRS agar by RAPD. The colonization rate of the different strains tested was very different. The RAPD-fast lysis method developed is simple and, with a low cost per assay, could also be applied to other food fermentations.     

A continuous dynamic beam model for swimming fish

When a fish swims in water, muscle contraction, controlled by the nervous system, interacts with the body tissues and the surrounding fluid to yield the observed movement pattern of the body. A continuous dynamic beam model describing the bending moment balance on the body for such an interaction during swimming has been established. In the model a linear visco-elastic assumption is made for the passive behaviour of internal tissues, skin and backbone, and the unsteady fluid force acting on the swimming body is calculated by the 3D waving plate theory. The body bending moment distribution due to the various components, in isolation and acting together, is analysed. The analysis is based on the saithe (Pollachius virens), a carangiform swimmer. The fluid reaction needs a bending moment of increasing amplitude towards the tail and near-standing wave behaviour on the rear-half of the body. The inertial movement of the fish results from a wave of bending moment with increasing amplitude along the body and a higher propagation speed than that of body bending. In particular, the fluid reaction, mainly designed for propulsion, can provide a considerable force to balance the local momentum change of the body and thereby reduce the power required from the muscle. The wave of passive visco-elastic bending moment, with an amplitude distribution peaking a little before the mid-point of the fish, travels with a speed close to that of body bending. The calculated muscle bending moment from the whole dynamic system has a wave speed almost the same as that observed for EMG-onset and a starting instant close to that of muscle activation, suggesting a consistent matching between the muscle activation pattern and the dynamic response of the system in steady swimming. A faster wave of muscle activation, with a variable phase relation between the strain and activation cycle, appears to be designed to fit the fluid reaction and, to a lesser extent, the body inertia, and is limited by the passive internal tissues. Higher active stress is required from caudal muscle, as predicted from experimental studies on fish muscle. In general, the active force development by muscle does not coincide with the propulsive force generation on the tail. The stiffer backbone may play a role in transmitting force and deformation to maintain and adjust the movement of the body and tail in water.