Interim report of the HELCOM CORESET project

damage to LMS (Köhler 1990, 1991). Salinity changes didn’t provoke signifi cant effects on LMS in fl ounder
(Broeg, unpublished results).
Using the cytochemical approach, temperature stress during the tissue incubation at 37°C has to be considered
when working with animals from subpolar and polar regions. For these animals, temperature stress
leads to a signifi cant decrease of LMS. In this case, temperature during incubation should not be higher than
20°C above the ambient temperature of the sampling location to avoid effects of too severe hyperthermia.
Ecological relevance
Lysosomal integrity is directly correlated with physiological scope for growth (SFG) and is also mechanistically
linked in terms of the processes of protein turnover (Allen and Moore 2004; Moore et al. 2006a). Ringwood
et al. (2004) have also shown that LMS in parent oysters is directly correlated with larval viability. It is also
inversely correlated with reproductive disorders in eelpout (Broeg & Lehtonen 2006). Finally, LMS is directly
correlated with diversity of macrobenthic organisms in an investigation in Langesund Fjord in Norway (Moore
et al. 2006b), and with parasite species diversity in fl ounder from the German Bight (Broeg et al. 1999).
Quality Assurance
Intercalibration exercises for LMS techniques have been carried out in the ICES/UNESCO-IOC-GEEP Bremerhaven
Research Workshop, the UNEP-MEDPOL programme, in the framework of the EU-project BEEP and
the BONUS+ project BEAST as well as for the neutral red retention method in the GEF Black Sea Environmental
Programme (Köhler et al. 1992; Lowe et al. 1992; Moore et al. 1998; Viarengo et al. 2000, BEEP
2004). The results from these operations indicated that both techniques could be used in the participating
laboratories in an effective manner with insignifi cant inter-laboratory variability.
Comparisons of the cytochemical and the neutral red retention techniques have been performed in fi sh liver
(ICES-IOC Bremerhaven Workshop, 1990) and in mussels experimentally exposed to PAHs (Lowe et al. 1995).
An AWI/Imare international workshop on “Histochemistry of lysosomal disorders as biomarkers in environmental
monitoring” in Bremerhaven, 2008, demonstrated good correspondence of results obtained by the
participants by applying various different assessments by computer assisted image analysis and light microscopy.
In 2010, an ICES\OSPAR Workshop on Lysosomal Stability Data Quality and Interpretation (WKLYS) has
been held in Alessandria, Italy (ICES 2010). This workshop concentrated on the NRR.
Guidelines for LMS procedures are published as ICES Times Series (Moore et al. 2004b), and in the UNEP/
Ramoge biomarker manual (UNEP 1999).
Assessment Criteria
Health status thresholds for NRR and cytochemical methods for LMS have been determined from data
based on numerous studies (Cajaraville et al. 2000; Moore et al. 2006a, Broeg et al. 2005, Broeg &
Lehtonen 2006).
LMS is a biophysical property of the bounding membrane of lysosomes and appears to be largely independent
of taxa. In all organisms tested to date, which includes protozoans, annelids (terrestrial and
marine), molluscs (freshwater and marine), crustaceans (terrestrial and aquatic), echinoderms and fi sh,
the absolute values for measurement of LMS (NRR and cytochemical method) are directly comparable.
Furthermore, measurements of this biomarker in animals from climatically and physically diverse terrestrial
and aquatic ecosystems also indicate that it is potentially a universal indicator of health status. For the cytochemical
method animals are considered to be healthy if the LMS is >20 minutes; stressed but compensating
if 10 minutes and severely stressed and probably exhibiting pathology if 120 minutes; stressed but compensating if 50 minutes and severely
stressed and probably exhibiting pathology if