Therefore, they should be required to develop highly sophisticated mechanisms to modulate maternal immune system during gestation to avoid immunological rejection, but little is known about this in non-mammalian vivipara. In Embiotocidae, a representative vivipar- ous family in teleosts, fetuses are retained in the ovarian cavity, with receiving maternal secretion, ovarian cavity fluid. Expecting that the fluid plays critical roles not only in nutrient supply but also in maintaining pregnancy, we purified and character- ized a major proteinaceous component of 51 kDa in the ovarian cavity fluid of Neoditrema ransonneti. On the basis of the N-terminal amino acid sequence determined, RACE-PCR was performed and a full length of cDNA was sequenced. The deduced amino acid sequence comprises 214 amino acids including 20 aa of signal peptides. This protein showed some unique characteristics: extremely high glycosylation (apparently more than 50%) and extraordinarily low pI (below 3.0). Sequence analysis revealed that this protein is a homolog of alpha-1- acid glycoprotein, a well-known immuno-modulating protein. This protein, named F-AGP, is produced in the liver but not in the ovary. The fact that F-AGP is specifically accumulated in the ovarian cavity indicates its selective transport from blood to the ovarian cavity, and, therefore, its special relevance to pregnancy in this species. doi:10.1016/j.cbpa.2008.05.166 8. Morphological examination and quantification of hepatic peroxisomes in the koala (Phascolarctos cinereus) by electron microscopy S.N.T. Ngo (University of South Australia, Australia; Charles Darwin University, Australia); R.A. McKinnon, I. Stupans, (University of South Australia, Australia) Peroxisomes are membrane bound cytoplasmic organelles that are involved in lipid metabolism and other biological functions. In rat and mouse, profound xenobiotic-induced peroxisome proliferation has been reported, with a marked increase in number and size of peroxisomes in liver parenchymal cells and induction of lipid metabolising enzymes, in particular cytochrome P450 CYP4A and peroxisomal β-oxidation acyl CoA oxidases (AOX). The present study investigates whether the previously observed higher hepatic CYP4A and AOX expression in the koala (Phascolarctos Cinereus), a unique Australian marsupial, compared with rat and human is associated with peroxisome proliferation. Visualisation and quanti- fication of peroxisomes were performed on liver samples from three koalas utilising transmission electron microscopy, with rat and bandicoot livers being used for comparative purposes. Numerous catalase positive peroxisomes, which clearly stand out by their black single membrane globular structures, were detected in all test ultra- thin sections from koala livers. A higher average number of peroxisomes per hepatocyte was observed for the koala, an obligate Eucalyptus feeder, compared with non-Eucalyptus feeders rat and bandicoot (counting of peroxisomes was performed on ≥ 50 hepatocytes/animal). No species differences in the average size of peroxisomes were detected (measurement of size was performed on ≥ 40 peroxisomes from different hepatocytes). This is the first morphological study examining hepatic peroxi- somes in an Australian marsupial. The results suggested that die- tary Eucalyptus constituents might possess peroxisome proliferating activities. doi:10.1016/j.cbpa.2008.05.167 9. Tributyltin (TBT) effect on the mitochondrial F 0 F 1 complex in the Mytilus galloprovincialis digestive gland A. Pagliarani, S. Nesci, V. Ventrella, F. Trombetti, M. Pirini, A.R. Borgatti (Alma Mater Studiorum Università di Bologna, Italia) The biochemical mechanisms involved in mollusc TBT toxicity remain at least in part unknown and TBT effects on mollusc mitochondria have up to now been poorly elucidated. The mitochon- drial oligomycin sensitive Mg-ATPase (OS-MgATPase), amounting to 80% of total mitochondrial MgATPase, was progressively depressed by increasing TBT concentrations up to a maximal inhibition of 70% at 0.7 μM TBT. Lineweaver Burk and Dixon plots exhibited the typical shape of an apparently non-competitive inhibition with respect to the ATP substrate with a Ki value of 0.23 μM TBT. MgATPase inhibition was partially released within the 0.7–5.0 μM TBT range up to a 70% activity recovery at 5.0 μM TBT. At higher TBT concentrations, OS-MgATPase activity appeared to be once again progressively depressed attaining nearly 100% enzyme inhibition at 30 μM TBT. A parallel increase in an oligomycin insensitive MgATPase (OI-MgATPase) activity was progres- sively observed. The hypothesis of a TBT-driven removal of F 1 subunit from the ATPase complex was excluded by incubation and centrifuga- tion procedures followed by convenient ATPase activity assays. Quercetin, a selective F 1 inhibitor, prevented the OI-MgATPase activity increase at higher than 5.0 μM TBT, thus suggesting that F 1 is not involved in TBT inhibitory mechanism and shouldering the hypothesis that F 0 subunit is the target of TBT. doi:10.1016/j.cbpa.2008.05.168 10. Linking responses at different levels of biological organization: The effect on biomarkers and life cycle traits in the earthworm Eisenia andrei exposed to copper-spiked soils A. Pasteris, N. Gambi, S. Guerrini, V. Bandini, A. Buscaroli, E. Dinelli, E. Fabbri (University of Bologna, Ravenna, Italy) Living organisms respond to chemicals at all the levels of biological organization, from molecules to ecosystems. In general, biochemical and physiological responses (molecules, cells, tissues) are triggered at low concentrations and after short exposure times, while ecological responses (life cycle traits, populations, communities, ecosystems) can be evidenced only at higher concentrations and after longer exposures. On the other hand, responses at the higher levels of organization are considered more relevant from an environmental point of view, as they often result in irreversible, severe damage to ecological systems. It is important to clarify the relationship between low-level and high-level responses to toxicants, particularly considering the increasing applica- tion of biomarkers as early warning systems to assess environmental pollution. In accordance with this view, effects of exposure to copper- spiked soils on life cycle endpoints were compared to effects on a battery of biomarkers in the earthworm Eisenia andrei. A natural clayey soil was spiked at seven copper concentrations, from 25 to 1600 mg/kg. Adult worms were exposed in laboratory to the spiked soils and to a control treatment (unspiked soil). Two sets of experimental container were prepared: the first to measure lisosomial stability, Ca 2+ ATP-ase activity, lipofuscin storage and metallotionein storage, after 10 d of exposure; the second to assess survival, growth and copper bioaccumulation after 28 d, and reproduction after 56 d of exposure. All the biological responses showed a clear concentration- S50 Abstracts / Comparative Biochemistry and Physiology, Part A 151 (2008) S48–S53