Recent Submissions

  • Using RIVPACS for studies on conservation and biodiversity

    Boon, P.J.; Wright, J.F.; Sutcliffe, D.W.; Furse, M.T. (Freshwater Biological Association, 2000)
    Invertebrate conservation relies not only on public support and political will, but also on possessing an adequate understanding of the distribution and ecology of invertebrate species and communities. In the UK, RIVPACS is making an important contribution to assessing the conservation importance of river invertebrate assemblages. So far, work has largely centred on using RIVPACS as an integral part of SERCON (System for Evaluating Rivers for Conservation), in which data collected using the standard RIVPACS method are interpreted with reference to conservation criteria such as species richness and representativeness. Applications of RIVPACS to other areas of conservation - whether providing information on the ecological requirements of rare species, monitoring the success of river restoration projects, or making broader assessments of sustainability - are probably more limited, but merit further examination. It is important to develop closer links between RIVPACS and techniques such as SERCON and RHS (River Habitat Survey) in order to maximise the benefit each can bring tostudies on conservation and biodiversity. It should also be recognised that there are limitations in transferring such systems to other countries where approaches to nature conservation may be very different.
  • The potential of RIVPACS for predicting the effects of environmental change

    Armitage, P.D.; Wright, J.F.; Sutcliffe, D.W.; Furse, M.T. (Freshwater Biological Association, 2000)
    RIVPACS has been used successfully for biological assessment of river water quality but its potential in forecasting the effects of environmental change has not been investigated. This study has shown that it is possible to simulate faunal changes in response to environmental disturbance, provided that the disturbance directly involves the environmental variables used in RIVPACS predictions. These variables relate to channel shape, discharge and substratum. Many impacts, particularly those associated with pollution, will not affect these variables and therefore RIVPACS cannot simulate the effects of pollution. RIVPACS was sensitive only to major changes in substratum. It was concluded that, because of the static nature of RIVPACS, it cannot respond to the dynamic effects and processes associated with environmental disturbance. Thus RIVPACS, while showing direction of change and indicating sensitive taxa, cannot be used to predict or forecast the effects of environmental impacts.
  • An introduction to RIVPACS

    Wright, J.F.; Wright, J.F.; Sutcliffe, D.W.; Furse, M.T. (Freshwater Biological Association, 2000)
    RIVPACS (River InVertebrate Prediction And Classification System) is a software package developed by the Institute of Freshwater Ecology (IFE). The primary application is to assess the biological quality of rivers within the UK. RIVPACS offers site-specific predictions of the macroinvertebrate fauna to be expected in the absence of major environmental stress. The expected fauna is derived by RIVPACS using a small suite of environmental characteristics. The biological evaluation is then obtained by comparing the fauna observed at the site with the expected fauna. RIVPACS also includes a site classification based on the macroinvertebrate fauna of the component reference sites. New sites, judged by their fauna to be of high biological quality, may be allocated to classification groups within the fixed RIVPACS classification. This has potential for evaluating sites for conservation. In this chapter, the origins and history of the RIVPACS approach are described, including major scientific and operational developments over the life of the project. RIVPACS III is described in detail and predictions at different taxonomic levels are demonstrated. The value of the reference dataset for river management and conservation is examined, and the chapter concludes with a brief consideration of some future challenges.
  • Detection of cryptosporidium oocysts in water and environmental concentrates

    Smith, H.V.; Sutcliffe, D.W. (Freshwater Biological Association, 1997)
    Whilst current methods for the isolation and enumeration of Cryptosporidium spp. oocysts in water have provided some insight into their occurrence and significance, they are regarded as being inefficient, variable and time-consuming, with much of the interpretation being left to the expertise of the analyst. Two expectations of novel developments are to reduce the variability and subjectivity associated with the isolation and identification of oocysts. Flocculation, immunomagnetisable and flow cytometric techniques, for concentrating oocysts from water samples, should prove more reliable than current methods, whilst the development of more avid and specific monoclonal antibodies in conjunction with the use of nuclear fluorochromes will aid identification. Further insight into the viability, taxonomy, species identification, infectivity and virulence of the parasite should be forthcoming through the use of techniques such as the polymerase chain reaction, in situ hybridisation and non-uniform alternating current electrical fields. Such information is necessary in order to enable microbiologists, epidemiologists, engineers, utility operators and regulators to assess the safety of a water supply, with respect to Cryptosporidium contamination, more effectively.
  • The diversity and ecological role of protozoa in fresh waters

    Finlay, B.J.; Sutcliffe, D.W. (Freshwater Biological Association, 1997)
    Protozoa feed on and regulate the abundance of most types of aquatic microorganisms, and they are an integral part of all aquatic microbial food webs. Being so small, aerobic protozoa thrive at low oxygen tensions, where they feed (largely unaffected by metazoan grazing) on the abundance of other microorganisms. In anaerobic environments, they are the only phagotrophic organisms, and they live in unique symbiotic consortia with methanogens, sulphate reducers and non-sulphur purple bacteria. The number of extant species of protozoa may be quite modest (the global number of ciliate species is estimated at 3000), and most of them probably have cosmopolitan distributions. This will undoubtedly make it easier to carry out further tasks, e.g. understanding the role of protozoan species diversity in the natural environment.
  • Immunodetection of planktonic algae

    Taylor, J.A.; Sutcliffe, D.W. (Freshwater Biological Association, 1997)
    Interest in the identification and characterisation of cyanobacteria and dinoflagellates in aquatic environments is increasing rapidly due to the perceived roles of these organisms in primary production and nuisance aspects in terms of water treatment and public health. Techniques for the identification and quantification of these organisms currently are limited, and the application of molecular approaches provides fundamental taxonomic information and techniques of practical value. Antigenic properties of algal cells may be useful taxonomic markers. Immunodetection techniques utilise the specificity of the antibody/antigen association as a probe for recognising and distinguishing between microorganisms according to their cell- surface chemistry. Immunofluorescent detection of unicellular cyanobacteria and dinoflagellates has been studied with success in marine and freshwater ecosystems and a range of techniques and results are presented and discussed. The most recent advances in the study of planktonic algae have come with the application of continuous flow cytometric methods (CFC). Flow cytometry makes use of the autofluorescence properties of the algal cells, which alone can be used to demonstrate their presence and permit their quantification in natural water samples. When used in conjunction with immunolabelling techniques, the potential of CFC analysis is broadened to study the serological/strain composition of plankters in natural populations. Changes in algal strains represented within and between waters over periods of time are reported and discussed, along with the ecological issues thus raised.
  • Recent progress in the identification and determination of freshwater phytoplankton in the natural environment

    Wilhelm, C.; Lohmann, C.; Sutcliffe, D.W. (Freshwater Biological Association, 1997)
    The biomass of the phytoplankton and its composition is one of the most important factors in water quality control. Determination of the phytoplankton assemblage is usually done by microscopic analysis (Utermöhl's method). Quantitative estimations of the biovolume, by cell counting and cell size measurements, are time-consuming and normally are not done in routine water quality control. Several alternatives have been tried: computer-based image analysis, spectral fluorescence signatures, flow cytometry and pigment fingerprinting aided by high performance liquid chromatography (HPLC). The latter method is based on the fact that each major algal group of taxa contains a specific carotenoid which can be used for identification and relative quantification of the taxa in the total assemblage. This article gives a brief comparative introduction to the different techniques available and presents some recent results obtained by HPLC-based pigment fingerprinting, applied to three lakes of different trophic status. The results show that this technique yields reliable results from different lake types and is a powerful tool for studying the distribution pattern of the phytoplankton community in relation to water depth. However, some restrictions should be taken into account for the interpretation of routine data.
  • Persistence of viable but non-culturable bacteria during the production and distribution of drinking water

    Cervantes, P.; Mennecart, V.; Robert, C.; de Roubin, M.R.; Joret, J.C.; Sutcliffe, D.W. (Freshwater Biological Association, 1997)
    The direct measurement of in situ respiring bacteria using 5-cyano-2,3-ditolyl tetrazolium chloride (CTC) shows that, especially for Gram-negative bacteria, large numbers of viable but non-culturable (VBNC) bacteria are present in finished water from a conventional water treatment plant, and the regrowth of bacteria along distribution networks can be seen rapidly by using this very sensitive technique. The level of bacterial inactivation with chlorine is much less important than has been previously supposed (based on experiments with non-injured laboratory strains of bacteria and classical culture techniques). Threshold values of VBNC bacteria leaving water treatment plants or regrowing along distribution systems have to be determined for better control of coliform regrowth and health- risks associated with the consumption of drinking water.
  • Biodiversity in drinking water distribution systems:a brief review

    Block, J.C.; Sibille, I.; Gatel, D.; Reasoner, D.J.; Lykins, B.; Clark, R.M.; Sutcliffe, D.W. (Freshwater Biological Association, 1997)
    In drinking water distribution systems, three groups of living organisms are usually found in the biofilm and circulating water: heterotrophic bacteria, free-living protozoa, and macro-invertebrates. Indirect evidence suggests that protozoa grazing in distribution systems can partially eliminate biomass production and accidental microbiological pollution. This paper examines the biodiversit in drinking water distribution systems.
  • Tastes and odours in potable water:perception versus reality

    Kelly, M.G.; Pomfret, J.R.; Sutcliffe, D.W. (Freshwater Biological Association, 1997)
    Tastes and odours are amongst the few water quality standards immediately apparent to a consumer and, as a result, account for most consumer complaints about water quality. Although taste and odour problems can arise from a great many sources, from an operational point of view they are either ”predictable” or ”unpredictable”. The former - which include problems related to actinomycete and algal growth - have a tendency to occur in certain types of water under certain combinations of conditions, whereas the latter - typically chemical spills - can occur anywhere. Long-term control is one option for predictable problems, although biomanipulation on a large scale has had utile success. Detection and avoidance is a more practicable option for both predictable and unpredictable problems, particularly if the distribution network can be serviced from other sources. Where these are not feasible, then water treatment, typically using activated carbon, is possible. In general there is a reasonable understanding of what compounds cause taste and odour problems, and how to treat these. An efficient taste and odour control programme therefore relies ultimately on good management of existing resources. However, a number of problems lie outside the remit of water supply companies and will require more fundamental regulation of activities in the catchment.
  • The health significance of heterotrophic bacteria in drinking water

    Lightfoot, N.; Sutcliffe, D.W. (Freshwater Biological Association, 1997)
    Tap water is not sterile; it contains organisms which grow in water distribution systems or inside taps and their fittings. The absence of known pathogenic bacteria is assured by the absence of the indicator organisms but concerns have been raised in the past few years that drinking water fulfilling the standards laid down in the EC Directive ECC 80/778 may still cause disease. These concerns have arisen from several sources: the fact that a cause has been identified in only half of all suspected waterborne outbreaks of disease; reports have suggested that heterotrophic bacteria possessing single pathogenic mechanisms such as haemolysin may cause disease; reports of heterotrophic organisms causing water contact diseases in hospitals. These concerns led to a reappraisal of the pathogenic potential of heteretrophic bacteria, by carrying out an extensive literature search and review commissioned by the UK Water Research Company. This research identified many papers showing an association between drinking water and heterotrophic bacteria but only very few reports of suspected waterborne disease associated with the heterotrophs. The organisms demonstrating potential to cause disease were species of Aeromonas and Yersinia, but typing of organisms identified in patients and isolated from the water revealed very few similarities. The potential of Aeromonas and Yersinia to cause waterborne disease is thought to be very low and the Communicable Disease Surveillance Centre database of laboratory infections due to these two genera of organisms was analysed to produce population-related incidences for each health region in England and Wales. Additionally a laboratory questionnaire revealed different levels of ascertainment of these two organisms in different laboratories of the Public Health Laboratory Service.
  • Viruses and drinking water

    Cartwright, R.Y.; Sutcliffe, D.W. (Freshwater Biological Association, 1997)
    There is no evidence to indicate that there is a risk of acquiring a virus infection through the consumption of properly treated drinking water, provided the integrity of the distribution system is maintained and there is no post-treatment contamination. The consumption of inadequately treated, untreated or post-treatment contaminated water is, however, associated with a risk of hepatitis A, hepatitis E and viral gastroenteritis. The use of the standard bacterial indicators for water monitoring provides an adequate safeguard against viral contamination.
  • Production of positive controls for calcivirus-specific PCR using recombinant baculiovirus technology

    Butcher, S.A.; Gould, E.A.; Sutcliffe, D.W. (Freshwater Biological Association, 1997)
    Recent advances in our knowledge of the genetic structure of human caliciviruses (HuCVs) and small round-structured viruses (SRSVs) have led to the development of polymerase chain reaction (PCR)-based molecular tests specific for these viruses. These methods have been developed to detect a number of human pathogenic viruses in environmental samples including water, sewage and shellfish. HuCVs and SRSVs are not culturable, and no animal model is currently available. Therefore there is no convenient method of preparing viruses for study or for reagent production. One problem facing those attempting to use PCR-based methods for the detection of HuCVs and SRSVs is the lack of a suitable positive control substrate. This is particularly important when screening complex samples in which the levels of inhibitors present may significantly interfere with amplificiation. Regions within the RNA polymerase regions of two genetically distinct human caliciviruses have been amplified and used to produce recombinant baculoviruses which express RNA corresponding to the calicivirus polymerase. This RNA is being investigated as a positive control substrate for PCR testing, using current diagnostic primer sets. Recombinant baculovirus technology will enable efficient and cost-effective production of large quantities of positive control RNA with a specific known genotype. We consider the development of these systems as essential for successful screening and monitoring applications.
  • Detection of specific bacteria in water: implications of survival strategy

    Pickup, R.W.; Rhodes, G.; Sutcliffe, D.W. (Freshwater Biological Association, 1997)
    It is widely recognised that conventional culture techniques may underestimate true viable bacterial numbers by several orders of magnitude. The basis of this discrepancy is that a culture in or on media of high nutrient concentration is highly selective (either through ”nutrient shock” or failure to provide vital co-factors) and decreases apparent diversity; thus it is unrepresentative of the natural community. In addition, the non-culturable but viable state (NCBV) is a strategy adopted by some bacteria as a response to environmental stress. The basis for the non-culturable state is that cells placed in conditions present in the environment cannot be recultured but can be shown to maintain their viability. Consequently, these cells would not be detected by standard water quality techniques that are based on culture. In the case of pathogens, it may explain outbreaks of disease in populations that have not come into contact with the pathogen. However, the NCBV state is difficult to attribute, due to the failure to distinguish between NCBV and non-viable cells. This article will describe experiences with the fish pathogen Aeromonas salmonicida subsp. salmonicida and the application of molecular techniques for its detection and physiological analysis.
  • Cultural methods of detection for microorganisms: recent advances and successes

    Watkins , J.; Jian, X.; Sutcliffe, D.W. (Freshwater Biological Association, 1997)
    Most microbiological methods require culture to allow organisms to recover or to selectively increase, and target organisms are identified by growth on specific agar media. Many cultural methods take several days to complete and even then the results require confirmation. Alternative techniques include the use of chromogenic and fluorogenic substances to identify bacteria as they are growing, selective capture using antibodies after short periods of growth, molecular techniques, and direct staining with or without flow cytometry for enumeration and identification. Future microbiologists may not use culture but depend on the use of specific probes and sophisticated detection systems.
  • The impact of molecular biology on assessment of water quality: advantages and limitations of current techniques

    Saunders, J.R.; Saunders, V.A.; Sutcliffe, D.W. (Freshwater Biological Association, 1997)
    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.
  • The microbiological quality of water: the nature of the problem

    Jones, J.G.; Sutcliffe, D.W. (Freshwater Biological Association, 1997)
    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.
  • Metallothionein as an indicator of water quality: assessment of the bioavailability of cadmium, copper, mercury and zinc in aquatic animals at the cellular level

    George, S.G.; Langston, W.J.; Sutcliffe, D.W. (Freshwater Biological Association, 1994)
    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

    Livingston, D.R.; Förlin, L.; George, S.G.; Sutcliffe, D.W. (Freshwater Biological Association, 1994)
    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.
  • Mussel eggs as indicators of mutagen exposure in coastal and estuarine environments

    Dixon, D.R.; Pascoe, P.L; Sutcliffe, D.W. (Freshwater Biological Association, 1994)
    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.

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