Interim report of the HELCOM CORESET project

3.10. Lysosomal membrane stability
Author: Katja Broeg, Doris Schiedek and Kari Lehtonen
ICES SGEH Biological Effects methods Background Documents
for the Baltic Sea region (ICES/OSPAR document from the ICES
SGIMC Report 2010, complemented and modifi ed by SGEH 2011
with information relevant for application in the Baltic Sea region)
Description of the indicator
Lysosomal functional integrity is a generic common target for environmental stressors in all eukaryotic
organisms from yeast and protozoans to humans (Cuervo 2004), that is evolutionarily highly conserved.
The stability of lysosomal membranes is a good diagnostic biomarker of individual health status (Allen and
Moore 2004; Broeg et al. 2005; Köhler et al. 1992, Lowe et al. 2006). Dysfunction of lysosomal processes
has been mechanistically linked with many aspects of pathology associated with toxicity and degenerative
diseases (Cuervo 2004; Köhler 2004; Köhler et al. 2002; Moore et al. 2006a, b, Broeg 2010). Lysosomes
are known to accumulate many metals and organic xenobiotics. Metals such as copper, cadmium and
mercury are known to induce lysosomal destabilisation in mussels (Viarengo et al. 1981, 1985a, b). LMS
is strongly correlated with the concentration of PAHs and PCBs in mussel tissue (Cajaraville et al. 2000;
Krishnakumar et al. 1994; Moore 1990; Moore et al. 2006a, b; Viarengo et al. 1992, Strand et al. 2009),
as well as organochlorines and PCB congeners in the liver of fi sh (Köhler et al. 2002, Broeg et al. 1992).
LMS of various species of mussel and fi sh from different climate zones clearly refl ect gradients of complex
mixtures of chemicals in water and sediments (Da Ros et al. 2002; Pisoni et al, 2004; Schiedek et al. 2006,
Barsiene et al. 2006; Sturve et al. 2005), point sources of pollution, single pollution events and accidents
(Garmendia et al. 2011; Einsporn et al. 2005; Broeg et al. 2002, Broeg et al. 2008, Nicholson and Lam,
2005) and also serves for the discovery of new “Hot Spots” of pollution (Bressling 2006; Moore et. al.
1998; 2004a).
LMS can also be used as a prognostic tool, able to predict liver damage and tumour progression in the
liver of various fi sh species (Broeg et al. 1999; Diamant et al. 1999; Köhler et al. 2002; Köhler 2004, Broeg
2010). Also hepatopancreatic degeneration in molluscs, coelomocyte damage in earthworms, enhanced
protein turnover as a result of radical attack on proteins, and energetic status an indicator of fi tness of
individuals within a population can be predicted (Allen & Moore 2004; Kirchin et al. 1992; Köhler et al.
2002; Moore et al. 2004a, 2006a; Nicholson & Lam 2005; Svendsen & Weeks 1995; Svendsen et al. 2004).
Recently it is tested for its prognostic potential with respect to reproductive disorders in amphipods in the
Baltic Sea. For eelpout, this prognostic potential could already been demonstrated. Low membrane stabilities
coincided with distinct reproductive disorders that indicated adverse effects at the population level
(Broeg and Lehtonen 2006).
Thus, LMS has been adopted by UNEP as part of the fi rst tier of techniques for assessing harmful impact
in the Mediterranean Pollution programme (MEDPOL Phase IV) and is also recommended as biomarker to
be included into the OSPAR Coordinated Environmental Monitoring Programme (pre-CEMP). LMS of blue
mussel from the Inner Danish waters and the Danish Belt Sea is part of the Danish monitoring programme
NOVANA since 2003 (Strand et al. 2009). It is also under consideration for the Swedish monitoring programme
(Granmo, pers. comm.). Methods applied to assess LMS are the Neutral Red Retention test (NRR)
on living cells like mussel haemocytes, and the cytochemical test on serial cryostat sections performed from
snap-frozen tissue. These methods are described in detail by Moore et al. (2004b). Currently a new method
is developed for the assessment of LMS in single tissue sections of small indicator species like amphipods
(Broeg and Schatz, in prep.).
Beside LMS, adverse lysosomal reactions to xenobiotic pollutants include swelling, lipidosis (pathological
accumulation of lipid), and lipofuscinosis (pathological accumulation of age/stress pigment) in molluscs but
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