Suitability of invertebrate and vertebrate cells in a portable impedance-based toxicity sensor: temperature mediated impacts on long-term survival.

Using ECIS (electric cell-substrate impedance sensing) to monitor the impedance of vertebrate cell monolayers provides a sensitive measure of toxicity for a wide range of chemical toxicants. One major limitation to using a cell-based sensor for chemical toxicant detection in the field is the difficulty in maintaining cell viability over extended periods of time prior to use. This research was performed to identify cell lines suitable for ECIS-based toxicity sensing under field conditions. A variety of invertebrate and vertebrate cell lines were screened for their abilities to be stored for extended periods of time on an enclosed fluidic biochip with minimal maintenance. Three of the ten cell lines screened exhibited favorable portability characteristics on the biochips. Interestingly, all three cell lines were derived from ectothermic vertebrates, and the storage temperature that allowed long-term cell survival on the enclosed fluidic biochips was also at the lower end of reported body temperature for the organism, suggesting that reduced cellular metabolism may be essential for longterm survival on the biochip. Future work with the ectothermic vertebrate cells will characterize their sensitivity to a wide range of chemical toxicants to determine if they are good candidates for use in a field portable toxicity sensor.

Response characteristics of an aquatic biomonitor used for rapid toxicity detection.

The response characteristics of an aquatic biomonitor that detects toxicity by monitoring changes in bluegill (Lepomis macrochirus Rafinesque) ventilatory and movement patterns were evaluated in single chemical laboratory studies at concentrations near the 96-h LC(50) concentration and at the EILATox-Oregon Workshop in sequential tests of multiple unknown samples. Baseline data collected prior to exposure allows each fish to serve as its own control. When at least 70% of exposed fish exhibit ventilatory or movement parameters significantly different from baseline observations, a group alarm is declared. In the laboratory studies, the aquatic biomonitor responded to the majority of chemicals at the 96-h lc(50) within an hour or less, although substantially higher response times were found for malathion and pentachlorophenol. Workshop tests of single chemical concentrations presented as blind samples were consistent with the laboratory test results. There were no alarms under control conditions in any test. Although data are limited, the aquatic biomonitor appears to respond more rapidly to chemicals causing membrane irritation, narcosis or polar narcosis than to acetylcholinesterase inhibitors or oxidative phosphorylation uncouplers. All four monitored parameters (ventilatory rate, cough rate, ventilatory depth and movement) contributed to identification of first alarms at acutely toxic levels. Understanding these response patterns can be useful in data interpretation for biomonitor applications such as surface water monitoring for watershed protection, wastewater treatment plant effluent monitoring or source water monitoring for drinking water protection.

Using higher organisms in biological early warning systems for real-time toxicity detection.

Many biological early warning systems (BEWS) have been developed in recent years that evaluate the physiological and behavioral responses of whole organisms to water quality. Using a fish ventilatory monitoring system developed at the US Army Centre for Environmental Health Research as an example, we illustrate the operation of a BEWS at a groundwater treatment facility. During a recent 12-month period, the fish ventilatory system was operational for 99% of the time that the treatment facility was on-line. Effluent-exposed fish responded as a group about 2.8% of the time. While some events were due to equipment problems or non-toxic water quality variations, the fish system did indicate effluent anomalies that were subsequently identified and corrected. The fish monitoring BEWS increased treatment facility engineers' awareness of effluent quality and provided an extra measure of assurance to regulators and the public. Many operational and practical considerations for whole organism BEWS are similar to those for cell- or tissue-based biosensors. An effective biomonitoring system may need to integrate the responses of several biological and chemical sensors to achieve desired operational goals. Future development of an 'electronic canary', analogous to the original canary in the coal mine, could draw upon advances in signal processing and communication to establish a network of sensors in a watershed and to provide useful real-time information on water quality.

Ecological risk assessment: implications of hormesis.

Hormesis is a widespread phenomenon across many taxa and chemicals, and, at the single species level, issues regarding the application of hormesis to human health and ecological risk assessment are similar. For example, convincing the public of a 'beneficial' effect of environmental chemicals may be problematic, and the design and analysis of laboratory studies may require modifications to detect hormesis. However, interpreting the significance of hormesis for even a single species in an ecological risk assessment can be complicated by considerations of competition with other species, predation effects, etc. Ecological risk assessments involve more than a single species; they may involve communities of hundreds or thousands of species as well as a range of ecological processes. Applying hormetic adjustments to threshold effect levels for chemicals derived from sensitivity distributions for a large number of species is impractical. For ecological risks, chemical stressors are frequently of lessor concern than physical stressors such as habitat alteration or biological stressors such as introduced species, but the relevance of hormesis to non-chemical stressors is unclear. Although ecological theories such as the intermediate disturbance hypothesis offer some intriguing similarities between chemical hormesis and hormetic-like responses resulting from physical disturbances, mechanistic explanations are lacking. Further exploration of the relevance of hormesis to ecological risk assessment is desirable. Aspects deserving additional attention include developing a better understanding of the hormetic effects of chemical mixtures, the relevance of hormesis to physical and biological stressors and the development of criteria for determining when hormesis is likely to be relevant to ecological risk assessments.

Animals as sentinels of human health hazards of environmental chemicals.

A workshop titled "Using Sentinel Species Data to Address the Potential Human Health Effects of Chemicals in the Environment," sponsored by the U.S. Army Center for Environmental Health Research, the National Center for Environmental Assessment of the EPA, and the Agency for Toxic Substances and Disease Registry, was held to consider the use of sentinel and surrogate animal species data for evaluating the potential human health effects of chemicals in the environment. The workshop took a broad view of the sentinel species concept, and included mammalian and nonmammalian species, companion animals, food animals, fish, amphibians, and other wildlife. Sentinel species data included observations of wild animals in field situations as well as experimental animal data. Workshop participants identified potential applications for sentinel species data derived from monitoring programs or serendipitous observations and explored the potential use of such information in human health hazard and risk assessments and for evaluating causes or mechanisms of effect. Although it is unlikely that sentinel species data will be used as the sole determinative factor in evaluating human health concerns, such data can be useful as for additional weight of evidence in a risk assessment, for providing early warning of situations requiring further study, or for monitoring the course of remedial activities. Attention was given to the factors impeding the application of sentinel species approaches and their acceptance in the scientific and regulatory communities. Workshop participants identified a number of critical research needs and opportunities for interagency collaboration that could help advance the use of sentinel species approaches.

The effects of lapsed time since feeding upon the toxicity of zinc to fish.

This study was undertaken to determine the effects of elapsed time from feeding to exposure upon the tolerance of goldfish (carassiu auratus l.) to zinc. Methods followed those in the 24 hour static bioassay of the Ohio River Valley Water Sanitation Commission (SMITH et al., 1973). Fish were fed for 25 minutes and at specific times after feeding (0, 1, 2, 4, 6, 12, 24, 72 hours) groups of 10 fish were placed into 18 liters of water containing a lethal concentration (100 mg/l) of zinc as ZnSO4. Time until death was recorded for individual fish. A total of 420 fish (excluding controls) were exposed to zinc in the course of seven experimental runs. Statistical analysis revealed a slight but non-significant increase in survival time as the interval between feeding and exposure to zinc increased. These variations in survival time do not appear to be of sufficient magnitude to support the standard acute bioassay requirement that fish not be fed for 24 or 48 hours prior to their exposure to a toxicant.