History of Biomarkers - Ecological Biomarkers: Indicators of Ecotoxicological Effects - page 30

invertebrates (Flammarion et al. 2002; Gagné et al. 2002; Charissou et al. 2004; Radetski et
al. 2004; Almeida et al. 2005; Cadet et al. 2005; Gagné et al. 2006; Nigro et al. 2006; Toyooka
and Ibuki 2007; Almeida et al. 2007).
2.5 Multibiomarker Approach
The multibiomarker approach to evaluate the environmental quality of water is recom-
mended by all specialists in ecotoxicology for the biological monitoring of the pollution of
the environment henceforth. However, a long way had to be traveled before this point was
reached, as discussed below.
At the University of Oslo in Norway, in August 1986 there took place a practical work-
shop on the biological effects of the pollutants under the auspices of the Group of Experts
on the Effects of Pollutants (GEEP) of the Intergovernmental Oceanographical Commission
of UNESCO. A special publication of the journal Marine Ecology Progress Series (volume
46, 1988) was devoted to the results of this workshop (GEEP Workshop). The workshop,
according to Bayne et al. (1988a), had several goals: (1) to evaluate methods covering a
broad spectrum from molecular approaches (biochemical level) to cellular and physiologi-
cal processes (levels of the cell and whole organism) to the structure of communities of
benthic organisms (community level); (2) the participants were to be researchers work-
ing on these subjects and interested in the measurement of the impact of pollution; (3)
biological samples have to be taken from a site known for its pollution gradient according
to a very precise protocol of sampling and analysis, and carried out during the work-
shop; (4) the participants to the workshop should follow a rigorous statistical model, that
is, without knowing the ranking of sites along the pollution gradient; (5) the biological
analyses carried out throughout the workshop were to be supplemented by meticulous
chemical analyses in order to evaluate the relationship between the levels of contamina-
tion and the biological responses. The collected material consisted of mussels ( M. edulis ),
crabs ( Carcinus maenas ), winkles ( L. littorea ), and flounders ( P. l e su s ), as well as sediments.
The Frier and Langesund fjords of the south of Norway were selected as sites of inter-
est because they showed a chemical gradient of contamination from the bottom of the
Frier fjord to the bay of Langesund. In the conclusions of the GEEP workshop, Bayne et
al. (1988b) emphasized the development of biochemical measurements responding to spe-
cific organic pollutants: PAHs and PCBs (P450 enzymes) or metals (MTs). These authors
concluded that measurements of EROD activity in the flounder P. l e su s give the clearest
and most sensitive response to the gradients of organic pollution. Later, an international
(European) program, Biological Effects of Environmental Pollution (BEEP) in Marine
Coastal Ecosystems, 2001-2004, was established with the aim of validating and intercali-
brating a battery of biomarkers of contaminant exposure and effects in selected indicator
species in the Mediterranean, the North Atlantic, and the Baltic Seas. One of the main
goals of the program was to set up a network of biomarker researchers around Europe and
to assess the applicability of biomarkers for different regions and species in the surround-
ing sea areas (Lehtonen et al. 2006). The selected biomarkers were specific biomarkers
(EROD, MT, AChE inhibition, FACs) but also histochemical biomarkers of toxic effects such
as neutral red accumulation showing a disturbed lipid metabolism or “general health”
biomarkers, reflecting cytotoxicity LMS and immunotoxicity [acid phosphatase activity
of macrophage aggregates (M-ACT) and macrophage aggregate size (M-AREA)] as well as