A tool for better management of water quality during periods of algal blooms: the case of the River Oise

Like other rivers in the Paris area, the Oise is subject to important seasonal algal blooms. This eutrophication generates notable problems for the production of drinking-water from a treatment plant on the river at Méry. A mathematical model has been developed to simulate variation in water quality in a pre-treatment storage basin, and another model is currently being adapted to model the River Oise. Integration of the two models should provide a comprehensive tool for predicting variations of phytoplankton and water-quality parameters associated with algal blooms. This will be a decision-aid for optimizing control of the treatment process for providing potable water.

A control of gas-vacuolate cyanobacteria

The cyanobacteria that cause problems in water supply are principally the colonial forms that are buoyed up by gas vesicles. The success of these organisms is due, in part, to their gas vesicles, which enable them to perform vertical migrations or to maintain themselves in the euphotic zone. The gas vesicles are also the root cause of the problems. In calm periods they cause the cyanobacteria to float to the water surface forming noxious scums, and they may prevent the colonies from sedimenting in water treatment plants. Gas vesicles are hollow, gas-filled structures; they are rigid but can be collapsed by the application of pressure. Their critical collapse pressure is influenced by their dimensions, which vary in different organisms. Gas vesicles are formed by the assembly of two types of protein, which determine their mechanical and physical properties. Methods for collapsing gas vesicles in natural populations of cyanobacteria will be considered. They may have application to the control of cyanobacteria in water supply.

The effects of stress on benthic algal communities

The effects of stress on both microalgal and macroalgal communities are considered. On one hand the contrasting approaches of studies of these two communities reflect intrinsic differences in plant size, longevity and ease of handling. On the other hand they reveal that biological monitoring of the potentially deleterious effects of man's activities has focused largely on freshwater environments in which macroalgae only occasionally dominate. Large conspicuous plants can be readily investigated as individuals, whereas it is virtually impossible to trace effects of stress on an individual cell of a vegetatively-reproducing microalga; a population approach is almost inevitably necessary. However, rapid turnover rates, a spectrum of ecological characteristics distributed between many taxa, and the potential for statistical analysis, have facilitated the use of microalgae in environmental impact studies. Failure to extend such investigations into marine systems rests as much on man's ability to ignore environmental deterioration until it affects his quality of life as on the visual dominance of seaweeds around our coasts. However, large gaps remain in our knowledge of both large and small algae; some reported community changes over time are suspect, and the causes of even blatant changes are not always apparent.

Use of macroinvertebrate communities to detect environmental stress in running waters

This paper presents an account of some current uses of RIVPACS (River Invertebrate Prediction and Classification System), a software package developed by the Institute of Freshwater Ecology (UK). Background information is also given on the unique data-set on which the system is based. Before discussing RIVPACS, we consider the range of environmental stresses encountered in flowing-water systems and some of the ways in which stresses may affect macroinvertebrate communities. The wide application and relevance of the RIVPACS approach was recognised when it was chosen as the biological method for use throughout the UK in the 1990 River Quality Survey (RQS). In the concluding section we list some lessons learnt both from the 1990 survey and from our own testing exercise, and we outline current developments which will lead to a new version of RIVPACS for use in the 1995 RQS.

From the individual to the community and beyond: water quality, stress indicators and key species in coastal waters

This review examines water quality and stress indicators at levels of organisation from the individual to the community and beyond by means of three case studies concentrating on rocky shores within the north-east Atlantic. Responses of dogwhelks (Nucella) to tributyltin pollution from antifouling paints is examined as the main case study. There are effects at the individual level (development of male sexual characteristics in the female leading to effective sterility) and population level (reduction in juveniles, few females and eventual population disappearance of dogwhelks in badly contaminated areas) but information on community level effects of dogwhelk demise is sparse. Such effects were simulated by dogwhelk removal experiments on well studied, moderately exposed ledges on shores on the Isle of Man. The removal of dogwhelks reduced the size and longevity of newly established Fucus clumps that had escaped grazing. Removal of dogwhelks also increased the likelihood of algal escapes. In a factorial experiment dogwhelks were shown to be less important than limpets \{Patella) in structuring communities but still had a significant modifying effect by increasing the probability of algal escapes. Community level responses to stress on rocky shores are then explored by reference to catastrophic impacts such as oil spills, using the Torrey Canyon as a case study. Recovery of the system in response to this major perturbation took between 10-15 years through a series of damped oscillations. The final case study is that of indicators of ecosystem level change in response to climate fluctuations, using ratios of northern \{Semibalanus balanoides) and southern (Chthamalus spp.) barnacles. Indices derived from counts on the shore show good correlations with inshore sea-water temperatures after a 2-year lag phase. The use of barnacles to measure offshore changes is reviewed. The discussion considers the use of bioindicators at various levels of organisation.

From organism to population: the role of life-history theory

The role of life-history theory in population and evolutionary analyses is outlined. In both cases general life histories can be analysed, but simpler life histories need fewer parameters for their description. The simplest case, of semelparous (breed-once-then-die) organisms, needs only three parameters: somatic growth rate, mortality rate and fecundity. This case is analysed in detail. If fecundity is fixed, population growth rate can be calculated direct from mortality rate and somatic growth rate, and isoclines on which population growth rate is constant can be drawn in a ”state space” with axes for mortality rate and somatic growth rate. In this space density-dependence is likely to result in a population trajectory from low density, when mortality rate is low and somatic growth rate is high and the population increases (positive population growth rate) to high density, after which the process reverses to return to low density. Possible effects of pollution on this system are discussed. The state-space approach allows direct population analysis of the twin effects of pollution and density on population growth rate. Evolutionary analysis uses related methods to identify likely evolutionary outcomes when an organism's genetic options are subject to trade-offs. The trade-off considered here is between somatic growth rate and mortality rate. Such a trade-off could arise because of an energy allocation trade-off if resources spent on personal defence (reducing mortality rate) are not available for somatic growth rate. The evolutionary implications of pollution acting on such a trade-off are outlined.

The connection between single species and ecosystems

Ecosystem level models are motivated by some combination of scientific and practical concerns. Those models motivated by practical concerns are likely to bear little historical relation to previous models. Mechanisms of interaction between particular species and their ecosystems vary enormously. Some species literally construct their own ecosystems. Others have more or less complex and important interactions with other species so that their presence or absence may alter the ecosystem. Prior information about the natural history of particular species can make ecosystem investigations quicker, cheaper, and more effective. The optimal resource for preparing to deal with the unlimited diversity of questions asked of ecologists would be a large' computerized databank of natural history observations for as many species as possible.

The ecological impact of different mechanisms of chronic sub-lethal toxicity on feeding and respiratory physiology

Sub-lethal toxicity tests, such as the scope-for-growth test, reveal simple relationships between measures of contaminant concentration and effect on respiratory and feeding physiology. Simple models are presented to investigate the potential impact of different mechanisms of chronic sub-lethal toxicity on these physiological processes. Since environmental quality is variable, even in unimpacted environments, toxicants may have differentially greater impacts in poor compared to higher quality environments. The models illustrate the implications of different degrees and mechanisms of toxicity in response to variability in the quality of the feeding environment, and variability in standard metabolic rate. The models suggest that the relationships between measured degrees of toxic stress, and the maintenance ration required to maintain zero scope-for-growth, may be highly nonlinear. In addition it may be possible to define critical levels of sub-lethal toxic effect above which no environment is of sufficient quality to permit prolonged survival.

Stress, shredders and streams: using gammarus energetics to assess water quality

This paper reviews the effectiveness of Gammarus scope for growth (SfG) as an indicator of water quality. In addition, the link between physiological changes and effects at higher levels of biological organisation is addressed. Exposure to a range of toxicants resulted in decreases in Gammarus SfG which were qualitatively and quantitatively correlated with subsequent reductions in growth and reproduction. Reductions in SfG were due principally to a decrease in energy intake (i.e. feeding rate) rather than an increase in energy expenditure. Gammarus pulex is an important shredder in many stream communities and stressed-induced reductions in its feeding activity were correlated with reductions in the processing of leaf litter by a semi-natural stream community. Hence, changes in the physiological energetics of Gammarus provide a general and sensitive indicator of stress which can be linked to effects at higher levels of biological organisation. Under long-term stress and hence prolonged reductions in SfG, animals may adapt by modifying their life-history strategies and producing fewer, larger offspring.

Effects of stress on the immune system of fish

The effects of stress on the immune system of various fish species including dab Limanda limanda, flounder Platichthys flesus, sea bass Dicentrarchus labrax and gobies Zosterisessor ophiocephalus, were investigated from laboratory and field experiments, using various assays to measure immunocompetence, correlated with histological and ultrastructural observations. Modulation of the immune system was demonstrated at tissue, cellular and biochemical levels following exposure to various stressors. The spleen somatic index was depressed in dab stressed in the laboratory and gobies collected from polluted sites in the Venice Lagoon. Differential blood cell counts consistently showed an increase in phagocytes and decrease in thrombocytes in fish exposed to various stressors. Phagocytic activity from spleen and kidney adherent cells was stimulated in dab stressed by transportation but depressed in fish exposed to chemical pollutants. Respiratory burst activity in phagocytic cells was also stimulated in stressed dab but depressed in sea bass exposed to cadmium. The results are discussed in relation to current concepts on stress in fish and the regulation of the immune system.

Mussel eggs as indicators of mutagen exposure in coastal and estuarine environments

The aim of this study was to develop a short-term genotoxicity assay for monitoring the marine environment for mutagens. Based on the developing eggs and embryos of the marine mussel Mytilus edulis, an important pollution indicator species, the test employs the sensitive sister chromatid exchange (SCE) technique as its end-point, and exploits the potential of mussel eggs to accumulate mutagenic pollutants from the surrounding sea water. Mussel eggs take up to 6 months to develop while in the gonad, which provides scope for DNA damage to be accumulated over an extended time interval; chromosome damage is subsequently visualised as SCEs in 2-cell-stage embryos after these have been spawned in the laboratory. Methods which measure biological responses to pollutant exposure are able to integrate all the factors (internal and external) which contribute to the exposure. The new cytogenetic assay allows the effects of adult exposure to be interpreted in cells destined to become part of the next generation.

Metallothionein as an indicator of water quality: assessment of the bioavailability of cadmium, copper, mercury and zinc in aquatic animals at the cellular level

The study of metallothioneins (MTs) has greatly improved our understanding of body burdens, metal storage and detoxification in aquatic organisms subjected to contamination by the toxic heavy metals, Cd, Cu, Hg and Zn. These studies have shown that in certain organisms MT status can be used to assess impact of these metals at the cellular level and, whilst validation is currently limited to a few examples, this stress response may be linked to higher levels of organisation, thus indicating its potential for environmental quality assessment. Molluscs, such as Mytilus spp., and several commonly occurring teleost species, are the most promising of the indicator species tested. Natural variability of MT levels caused by the organism's size, condition, age, position in the sexual cycle, temperature and various stressors, can lead to difficulties in interpretation of field data as a definitive response-indicator of metal contamination unless a critical appraisal of these variables is available. From laboratory and field studies these data are almost complete for teleost fish. Whilst for molluscs much of this information is lacking, when suitable controls are utilised and MT measurements are combined with observations of metal partitioning, current studies indicate that they are nevertheless a powerful tool in the interpretation of impact, and may prove useful in water quality assessment.

Molecular biomarkers and toxic consequences of impact by organic pollution in aquatic organisms

Organic contaminants are readily bioaccumulated by aquatic organisms. Exposure to and toxic effects of contaminants can be measured in terms of the biochemical responses of the organisms (i.e. molecular biomarkers). The hepatic biotransformation enzyme cytochrome P4501A (CYP1A) in vertebrates is specifically induced by organic contaminants such as aromatic hydrocarbons, PCBs and dioxins, and is involved in chemical carcinogenesis via catalysis of the covalent binding of organic contaminants to DNA (DNA-adducts). Hepatic CYP1A induction has been used extensively and successfully as a biomarker of organic contaminant exposure in fish. Fewer but equally encouraging studies in fish have used hepatic bulky, hydrophobic DNA-adducts as biomarkers of organic contaminant damage. Much less is known of the situation in marine invertebrates, but a CYPlA-like enzyme with limited inducibility and some potential for biomarker application is indicated. Stimulation of reactive oxygen species (ROS) production is another potential mechanism of organic contaminant-mediated DNA and other damage in aquatic organisms. A combination of antioxidant (enzymes, scavengers) and pro-oxidant (oxidised DNA bases, lipid peroxidation) measurements may have potential as a biomarker of organic contaminant exposure (particularly those chemicals which do not induce CYP1A) and/or oxidative stress, but more studies are required. Both CYP1A- and ROS-mediated toxicity are indicated to result in higher order deleterious effects, including cancer and other aspects of animal fitness.

The microbiological quality of water: the nature of the problem

Improvements in methods for the detection and enumeration of microbes in water, particularly the application of techniques of molecular biology, have highlighted shortcomings in the ”standard methods” for assessing water quality. Higher expectations from the consumer and increased publicity associated with pollution incidents can lead to an uncoupling of the cycle which links methodological development with standard-setting and legislation. The new methodology has also highlighted problems within the water cycle, related to the introduction, growth and metabolism of microbes. A greater understanding of the true diversity of the microbial community and the ability to transmit genetic information within aquatic systems ensures that the subject of this symposium and volume provides an ideal forum to discuss the problems encountered by both researcher and practitioner.

The impact of molecular biology on assessment of water quality: advantages and limitations of current techniques

The advent of molecular biology has had a dramatic impact on all aspects of biology, not least applied microbial ecology. Microbiological testing of water has traditionally depended largely on culture techniques. Growing understanding that only a small proportion of microbial species are culturable, and that many microorganisms may attain a viable but non-culturable state, has promoted the development of novel approaches to monitoring pathogens in the environment. This has been paralleled by an increased awareness of the surprising genetic diversity of natural microbial populations. By targeting gene sequences that are specific for particular microorganisms, for example genes that encode diagnostic enzymes, or species-specific domains of conserved genes such as 16S ribosomal RNA coding sequences (rrn genes), the problems of culture can be avoided. Technical developments, notably in the area of in vitro amplification of DNA using the polymerase chain reaction (PCR), now permit routine detection and identification of specific microorganisms, even when present in very low numbers. Although the techniques of molecular biology have provided some very powerful tools for environmental microbiology, it should not be forgotten that these have their own drawbacks and biases in sampling. For example, molecular techniques are dependent on efficient lysis and recovery of nucleic acids from both vegetative forms and spores of microbial species that may differ radically when growing in the laboratory compared with the natural environment. Furthermore, PCR amplification can introduce its own bias depending on the nature of the oligonucleotide primers utilised. However, despite these potential caveats, it seems likely that a molecular biological approach, particularly with its potential for automation, will provide the mainstay of diagnostic technology for the foreseeable future.