10.1.
Environmental Risk Assessment and Mitigation of Hazardous Chemical Contamination
Business Plan
LABORATORY FOR MOLECULAR ECOTOXICOLOGY (LME)
LABORATORY FOR BIOGEOCHEMISTRY OF ORGANIC COMPOUNDS (LBOC)
M. Ahel, D. Hršak, M. Picer and T. Smital
Vision
The main idea is to form a regional center for risk assessment and mitigation of hazardous chemical contamination in the marine and terrestrial ecosystems. A comprehensive assessment of environmental risk associated with the increasing use of a large variety of toxic chemical contaminants requires an extremely high level of multidisciplinary expertise, which can be found exclusively in specialized interdisciplinary-oriented scientific institutions. Moreover, a truly objective judgment of the environmental issues can be achieved only through fully independent institutes that are capable to respond to growing public demands for a safe and healthy environment.
Mission
The mission of the future Center comprises research, postgraduate education and performing of services in the field of environmental protection, including monitoring, ecotoxicological assessment and life cycle analysis of harmful substances, as well as developing strategies for the prevention and mitigation of hazardous chemical pollution. It also envisages a market-oriented dissemination of the results of fundamental and applied research to various users, such as governmental agencies, local authorities and industry in order to promote a scientifically-based approach to the detection, assessment and mitigation of risks associated with usage and disposal of hazardous substances.
Objectives
Research:
§ Systematic investigation of hazardous chemical contamination in the marine and freshwater ecosystems as a basis for sustainable development of the Republic of Croatia
§ Development of new highly specific methods for the determination of xenobiotic compounds in environmental samples
§ Investigation of detoxification mechanisms in aquatic organisms and development of new biomarkers to detect hazardous chemical contamination (e.g. multixenobiotic resistence MXR)
§ Evaluation of the combined use of chemical and biomarker approach in the ecological risk assessment
§ Study of microorganisms as catalysts in biotransformation processes and cycling of elements in the biosphere
§ Investigation of metabolic pathways and biotransformation kinetics of ecotoxicologically relevant chemical contaminants, with the emphasis on highly toxic, genotoxic and endocrine disrupting substances
Education:
§ Participation in postgraduate studies in marine sciences (specialized courses, execution of Ms and PHD theses)
§ Participation in postgraduate studies in environmental protection (specialized courses, execution of Ms and PHD theses)
§ Teaching and providing specialized training courses in environmental analytical chemistry, ecotoxicology, environmental microbiology, implementation of environmental directives
Development of new measurement techniques:
§ Introduction of the most reliable state-of-the-art methodologies for the determination the prominent pollutant classes foreseen by the Croatian regulations and EU Water Directive
§ Becoming a center of excellence for the analysis of war-related hazardous pollution in soil and aquatic systems
§ Obtaining ISO certification for the standardized methods for the determination of the most common xenobiotic pollutants such as PAH and organo-halogen compounds
§ Development of suitable in vitro and in vivo bioassays for the determination of transport activity of specific proteins included in MXR phenomenon;
§ Development of appropriate methods and statistical models for the determination of additive, antagonistic and synergistic effects of binary (or even more complex) combination of pollutants
§ Introduction, standardisation and intercalibration of in vitro and in vivo methods for the detection of xenoestrogenic effects of environmentally relevant pollutants; development of molecular biology techniques (PCR, DGGE and in situ rRNA hybridization) for the characterisation of detoxification mechanisms in aquatic organisms and for the detection and identification of active microorganisms, participating in biotransformation processes
§ Validation of the reference methods for biodegradability and ecotoxicity testing
Services:
§ Monitoring of various pollutants and exposure biomarkers in the framework of national projects supported by Ministries and governmental agencies
§ Expert assistance in accidental situations
§ Authorised reference laboratory for the assessment of ecotoxicity, biodegradability and other biological parameters in wastewaters and natural waters
§ Environmental impact studies and consulting
Technology development and support:
§ Participation in the development of biotechnological processes for the treatment of xenobiotic substances in wastewater and in situ biotransformation. (e.g designing a treatment plant for purification of wastewaters from the herbicide production)
§ Development of new procedures for the advanced wastewater treatment based on membrane technologies (e.g. treatment of landfill leachate containing biorefractory pharamaceuticals)
Background
The co-workers of the future Center presently work in 2 Laboratories of the Center for Marine and Environmental Research, Laboratory for biogeochemistry of organic compounds (LBOC) and Laboratory for molecular ecotoxicology (LME). The Laboratories consist, in fact, of 4 autonomous research groups with distinct goals and funding resources. With respect to the proposed new association, the groups have complementary structures including environmental analytical chemistry, biogeochemistry, molecular ecotoxicology, environmental microbiology and biotechnology. All these disciplines form a promising network regarding the high requirements, which are needed to cope with the complex issues of environmental risk assessment. Besides the fundamental research, which considered as the only solid basis for all intended applications, the expertise gathered around the proposal allows the initiation of activities towards development and support of new technologies for wastewater treatment and in situ remediation. Besides an adequate scientific background, each of individual groups has a long experience in applying their fundamental research to the practical problems in the environment.
The Group for organic pollutant analysis was founded in 1974 with the main task to initiate gas chromatographic methods for the analysis of organochlorine pollutants in the marine environment. The members of the group have gradually broadened their interest to other pollutant classes, such as hydrocarbons, methyl-mercury, volatile halogenated hydrocarbons, anionic and nonionic surfactants and their metabolites, triazine herbicides and pharmaceutical chemicals, evolving into two separate groups (LME-analytical and LBOC-biogeochemical). Research activity of the LME-analytical group can be divided into two main branches: a) development of methods for the analysis of organic pollutants in environmental samples and b) application the developed methods in various field studies aimed at determining exposure levels and fate of selected pollutants in soil, marine and freshwater environments. Methods for analysis mineral oil and chlorinated hydrocarbons have been successfully intercalibrated in numerous domestic and international intercalibration exercises.
Research programme of the LBOC-biogeochemical group comprises systematic investigation of biogeochemical behaviour and fate of biogenic and anthropogenic organic compounds in different freshwater and marine environments using highly specific analytical techniques such as high-resolution gas chromatography, high-performance liquid chromatography and mass spectrometry. Regarding biogenic compounds, a special attention was paid to the natural compounds which play a major role in organic carbon cycling (e.g. photosynthetic pigments and carbohydrates), while for xenobiotic chemicals emphasis was on anionic and nonionic surfactants and emerging pollutants having pharmaceutical origin. The research programme includes studies of environmental processes, which govern the fate of organic compounds in real systems with a special emphasis on biodegradation. Intensive field studies have been carried out in the large variety of aquatic environments, ranging from marine environments to the alluvial aquifers. Frequent sampling campaigns in the marine environments are mainly focussed on the shelf and estuarine areas influenced by increased freshwater inputs (e.g. northern Adriatic and Krka River and Zrmanja River estuaries) with the emphasis on eutrophication-related phenomena.
The members of both groups participated in UNEP MED POL Projects of Monitoring the Adriatic Sea (Phase I. Phase II and Phase III) and former Yugoslav-Italian cooperation in monitoring the Adriatic Sea. The Group have taken part in numerous ecological studies of the various parts of the Adriatic environment andfresh water environment in Croatia. During the 1991-1995 war in Croatia, and especially after the “Oluja“-action in 1995, the LME-group has been intensively involved in the investigation of the ecological consequences of the war damages in the karst area of Croatia.
The members of analytical/biogeochemical groups published more than 200 scientific and professional papers dealing with the analytical determination environmental fate of various organic and organo-metallic pollutants in soil and water, earning more than 1600 citations.The average impact factor per paper published in CC-journals (being an indicator of the quality of journals) is 1.286 The total number of citations received by the researchers of our laboratory (period 1978-2001) is about 1400 (including some cross-citations In terms of scientific productivity in the period 1996-2001, the number of papers published in CC and SCI-journals per faculty is about 5.
The basic goal of the Laboratory for molecular ecotoxicology (LME) is to improve Ecological Risk Assessment by discovering and establishing new, better and more relevant biomarkers as important indicators of environmental quality.
The research group is primarily focused on the identification of detoxification and/or defense mechanisms that are present in taxonomically various classes of marine and freshwater organisms. The work comprises explanation of the physiological and biochemical relationships between the activity of detoxification mechanisms and the level of environmental pollution with the aim to develop and standardize biomarkers that would allow the measurement of the activity of these defense mechanisms on the molecular, and even more important on the sublethal level. This approach offers important information on biologically relevant exposure levels and effect of environmental pollutants and provides a scientific basis for both national and international legislation as well as for rational spending of funds assigned to environmental management and/or protection.
In the last 5 years LME published 24 papers in SCI quoted journals, with the average impact factor per paper of 1.264 (Note: average impact factor of journals listed in Environmental Sciences is ~ 0.737) and average citations per paper of 6.4. Average number of papers per faculty is about 4. One patent was filed. The LME-ecotoxicology research group discovered the presence of a new defense mechanism inherently present in aquatic organisms. The discovery of this defense system, hereafter in aquatic organisms named Multixenobiotic Resistance Mechanism (MXR), had exceptional ecotoxicological significance, especially after investigations that demonstrated that MXR might be inhibited by specific chemicals, so-called MXR inhibitors that are also present in aquatic environment. Therefore, MXR-inhibitors may be considered as a new potentially dangerous class of chemicals. During the last five-year period LME actively participated in several ecological and biomonitoring studies and as a consequence of this MXR mechanism was recently classified as a promising (“under development”) biomarker within international (US EPA, ICES, UNEP MED POL) biomonitoring programs.
The main research interest of the research group for Environmental microbiology and biodegradation (LBOC- environmental microbiology) is in basic principles governing the biotransformation of complex synthetic organic compounds (xenobiotics) with a special emphasis on microorganisms as essential factor (catalysts) in biotransformation processes. The research activities are focused on the enrichment, isolation, identification and characterization of bacterial communities from various pristine and polluted environments with the aim to study their roles in the transformation of complex synthetic organic compounds. This research is a prerequisite for better understanding the principles of the biogeochemical cycling of elements in the biosphere and is crucial for opening the possibility to influence the rate and the extent of xenobiotics transformation. Furthermore, this research provides a basis for the optimisation of the existing and development of new biotechnological procedures for the remediation of chemically-polluted sites as well as for improving the efficiency of traditional treatment of domestic and industrial wastewaters that regularly contain various xenobiotic compounds.Besides these basic investigations, a significant part of the research has been carried out in the framework of applied projects. Currently, an innovative-technological project is in progress with the main goal for designing the biotechnological procedure for the treatment of wastewater from atrazine production. Specific goals of these projects are possible improvements in maximizing atrazine recovery from technological wastewater and to meet the new, stringent discharge standards (minimize raw water use, wastewater generation and pollution of natural waters).
All research groups involved in this proposal have had an intensive co-operation with many internationally recognized laboratories, including those in EU and USA. The staff actively participated in several international projects, in some of them acting as co-proposers and scientific co-ordinators. Very recently, the leader of the LME-analytical group has been accepted as the scientific co-ordinator of a project on “Assessment of the selected POPs (PCBs, PCDDs/F, OCPs) in the athmosphere and water ecosystems from the waste materials generated by warfare in area of former Yugoslavia”, which is being realised within the EU Fifth framework INCO Copernicus Programme, involving 12 institutions from Croatia (3 institutions), Slovenia, Bosnia and Herzegovina, Kosovo, FR Yugoslavia (3 institutions), Austria, Czech Republic and Greece.
Current situation
LME-Ecotoxicology group
Staff:
§ The Lab staff include 5 researchers (educated in ecology, molecular biology and ecotoxicology) + 1 technician;
Existing infrastructure:
§ Lab space (6 rooms, 136 m2);
§ Supraspeed centrifuge Sorvall RC 28S (2 x) + other centriguges;
§ Microplate fluorimeter Fluorolite 1000 (Dynatech); Microplate photometer Anthos HT III (Anthos);
§ Incubator, LaminAir, inverse microscope (Olympus) and other cell culture equipment (Heraeus);
§ Colony counter and other equipment for Ames microsomal test;
§ Freezers (-80°C, 2 x), liquid nitrogen containers;
§ "Wet" lab (basins, experimental aquaria, flow of dechlorineted tap water;
§ Rotary evaporator, sterilizators, autoclav, balances, etc.
Major projects/ revenues
§ ”Multixenobiotic Resistance Mechanism as a Biomarker of Environmental Quality”, 2002-2004) – Project supported by the Ministry of Science and Technology of The Republic of Croatia - estimated budget ~ 22.000 $ (EUR/year);
§ “Projekt Jadran” - Systematic investigation of the Adriatic Sea as a basis for sustainable development of the Republic of Croatia (a project funded by Ministry of Science and Technology) ) – estimated budget ~ 11.000 $ (EUR/year);
§ ”An integrated environmental monitoring system for Croatian freshwater, estuarine and coastal marine areas”, (2002-2004) – Bilateral Project supported by the Research Council of Norway and the Ministry of Science and Technology of the Republic of Croatia – our budget ~ 28.000 $ (EUR)/year;
§ ”Ecological study of gas fields in the northern Adriatic”, (2001.-) – Pilot project supported by INA (Croatian oil and gas company) – estimated budget ~ 10.000 $ (EUR)/year.