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The basic morphological, ethological, and physiological-biochemical adaptations of Monodonta turbinata to survival in the littoral zone were investigated in this work. Quantitative estimation of myoglobin content in radular tissues of mollusks inhabiting the Mediterranean Sea Basin has been carried out.

A regression model is proposed for studying the relationship between trace metals concentrations and weights of individuals of Monodonta turbinata (Born), a gastropod mollusc used as a trace metal biomonitor of marine coastal areas. The model herein presented was developed including sites as reference factors and showed very good correlations. The prevision model of contamination gave good results for this kind of pollutants, and it can be used as a tool for monitoring programmes.

Morphological and biochemical adaptations of two Mediterranean mollusks Gastrana fragilis and Monodonta turbinata to living in the littoral zone which is most exposed to abiotic factors are analyzed. Hemoglobin favoring to a certain extent their survival under constantly changing environmental conditions was quantified in the mollusks.

Mehanizam multiksenobiotičke otpornosti (MXR) u vodenih organizama predstavlja prvu liniju obrane od potencijalno toksičnih ksenobiotika iz okoliša. MXR je posredovan transportnom aktivnošću transmembranskog P-glikoproteina (Pgp) koji aktivno izbacuje ksenobiotike iz stanice/organizma. Međutim, tvari koje nazivamo MXR-inhibitorima ili kemosenzitizatorima mogu blokirati aktivnost Pgp-a uzrokujući povećanu akumulaciju ksenobiotika i mogući toksični učinak. Temeljem ovih saznanja cilj ovog istraživanja bio je procijeniti vrijednost in vivo mjerenja MXR-aktivnosti kao parametra u procesu procjene okolišnog rizika (POR). MXR-aktivnost mjerili smo metodom akumulacije ili izbacivanja fluorescentnog modelnog MXR-supstrata rodamina B u/iz škrga odabranih pokusnih organizama (slatkovodni i morski školjkaši i puževi). Naši rezultati pokazali su: (1) da je MXR genetski urođen svim ispitivanim vodenim organizmima, ali da postoje značajne interspecijske razlike u MXR-aktivnosti. Razina MXR-aktivnosti podudara se s tolerancijom pojedinih vrsta na organsko zagađenje. Među ispitivanim vrstama kao najpogodniji pokusni organizmi za biomonitoring u svrhu mjerenja MXR-aktivnosti pokazali su se slatkovodni školjkaš Dreissena polymorpha, odnosno morski školjkaš Mytilus galloprovincialis; (2) određivanje koncentracije MXR-inhibitora u uzorcima vode s referentnih zagađenih i nezagađenih lokacija pokazalo je značajnu korelaciju sa stupnjem zagađenja. Industrijski i komunalno opterećene vode dokazano imaju i višu koncentraciju MXR-inhibitora, a prisustvo takvih tvari može u potpunosti poništiti zaštitni učinak MXR-mehanizma; (3) MXR-mehanizam je vrlo elastičan, tj. inducibilan i deinducibilan. Međutim, mjerenje indukcije MXR-aktivnosti kao biomarkera izloženosti ksenobioticima za sada je teško primjenjivo u procesu POR-a. Naime, osjetljivost opisanih metoda u ovu svrhu nije na odgovarajućoj razini, a brojni čimbenici otežavaju interpretaciju rezultata. Na temelju ovih rezultata zaključujemo da je ekotoksikološki značaj prisustva i funkcije MXR-mehanizma u vodenih organizama neupitan. Istraživanja ključnih molekularno bioloških značajki MXR-fenomena u vodenih organizama treba nastaviti, pa i proširiti razjašnjavanjem uloge drugih sličnih transportnih proteina, a određivanje koncentracije MXR-inhibitora kao donedavno nepoznate skupine okolišno opasnih tvari može značajno unaprijediti proces POR-a, te stoga treba postati sastavni dio kako svake ekološke studije, tako i zakonske regulative.

Protamine of the archaeogastropod mollusc Monodonta turbinata has been isolated and characterized. With a mass of 13,476 Da, it is the largest known prolamine. Amino acid sequence of this protamine (106 residues) was established from data provided by automated sequence analysis and mass spectrometry of the protein and of its fragments. The primary structure of the NH2-terminal region exhibits repetitive sequence motifs “Basic-Ser” (mainly R-S) and both central and COOH-terminal regions are composed by arginine clusters. The amino acid sequence of Monodonta turbinata protamine shows structural similarities with other protamines from invertebrates and from birds and mammals.

Many marine organisms inherently express the presence and function of a new defence mechanism, termed multixenobiotic resistance mechanism (MXRM) because of its similarity to multidrug resistance (MDR) found in tumor cell lines resistant to chemotherapeutic drugs. However, previously no information was available on a possible induction of the activity of MXRM in organisms living at polluted sites. The purpose of the present study was to demonstrate the inducibility of this defence mechanism in the marine snail Monodonta turbinata, an organism known to inhabit a wide range of environmental conditions existing along a pollution gradient. Specimens of M. turbinata used in the present study were collected from March 1993 through March 1994 near Rovinj, Croatia. The accumulation of generally labelled 3H-vincristine (3H-VCR) in the gills of snails living at an unpolluted site was 67% higher and very sensitive to verapamil, an inhibitor of P-glycoprotein activity in comparison to snails living at a polluted site. The accumulation of vincristine (VCR) in snails from a polluted site was reduced and insensitive to verapamil due to the induced state of P-glycoprotein, as was demonstrated immunochemically by the enhanced concentration of a 140 kDa protein cross-reacting with the polyclonal antibodies raised against mammalian P-glycoprotein. MXRM could be induced in snails from the unpolluted site to the level found in snails living at a polluted site either by transplantation for 3 d to a polluted site, or by exposure for 3 d to sea water polluted with Diesel-2 oil (hydrocarbon concentration equivalent to 50 ppb of Kuwait oil). Discovery of the inducibility of this new defence mechanism in aquatic organisms may have important implications in ecotoxicology, as was demonstrated by a 104% enhanced accumulation of VCR in the presence of the MXRM-inhibitor, the so-called “chemosensitizer”. Besides, a striking difference in the levels of MXRM-elements (the sensitivity of the accumulation rate of xenobiotics to verapamil, immunochemical expression of P-glycoprotein, and the sensitivity of the binding of xenobiotics on membrane vesicles to verapamil) found between specimens living at the polluted and unpolluted sites offers a new molecular biomarker for exposure to pollutants.

This work underlines the importance of the statistical procedures necessary for the significant assessment of analytical data of heavy metals derived from marine organisms (biomonitors) collected at five stations in Ustica at the Southern Tyrrhenian Sea. The statistical method described indicates that, where the differences among the averages of Pb, Cu and Zn concentrations in the molluscs of the sites analysed tested with ANOVA are significant, it is necessary to conduct multiple comparison tests in order to determine which average is significantly different from the other. Using the method described, it is possible to indicate which site or sites is/are more contaminated in comparison with the other sites. This way it is possible to make a preliminary evaluation of the basal levels of trace metals present in the reference marine ecosystem under study.

The trochid snail Monodonta turbinata was exposed to various concentrations of mercury and cadmium salts at different temperatures. Survival was significantly affected by salt concentration and by temperature, as well as by the interaction between them, so that the toxicities of both mercury and cadmium salts were found to be enhanced at higher temperatures. The ecological significance of these results on this Mediterranean littoral trochid is discussed.

The effects of zinc and copper pollution on the allozymic variation of phosphoglucose isomerase (Pgi) genotypes were tested in the Mediterranean marine gastropods Monodonta turbinata and M. turbiformis in 26 laboratory tests involving 2 081 adult individuals, with 545 survivors. Our results indicate differential survivorship of electrophoretical Pgi allozyme genotypes for both pollutants. Zinc pollution selected against the allozyme genotype SS in M. turbinata, whereas no differential survivorship was observed in M. turbiformis which lacks this allozyme. Copper pollution selected against the MM genotype in both species. These results are inconsistent with the neutral theory of allozyme polymorphisms and appear to reflect the adaptive nature of some Pgi genotypes in these marine gastropods.

The sperm cells ofPatella coerulea (Patellacea),Monodonta turbinata, andGibbula tumida (Trochacea) were investigated by means of transmission electron microscopy. They belong to the primitive type (sensu Franzen) and have more features in common with primitive Bivalvia sperms than with Neritacea. Their head contains an apical acrosome and a roundish nucleus followed by 4 or 5 mitochondria and a centriolar apparatus which consists of two centrioles, one of which bears a flagellum. The sperm cells ofMonodonta andGibbula are very similar to each other and differ mainly in size;Patella exhibits more differences (very small acrosome, subacrosomal space, variable number of spherical mitochondria (origin of spermic dimorphism ?). The development of the sperm cells shows no peculiarities.

Zonal oscillations of Monodonta turbinata (Born) were monitored with an actographic device which precluded clustering behaviour. Unfed snails maintained their cyclic pattern of behaviour for up to 8 days under a light-dark cycle which simulated the natural one. Under constant conditions of light or dark, however, the snails ceased migration and occupied a zonal position typical of day and night, respectively. Experiments in diffuse light and with light from below the floor of the experimental tank showed that the downward migration of M. turbinata in the daytime depends on positive geotaxis combined with negative phototaxis whilst the upward migration at night depends on a negative geotaxis. This mechanism is similar to that described in other littoral molluscs. There was no evidence of an endogenous control of rhythmic zonal activity.

Samples of Patella coerulea (Linnaeus), Brachydontes variabilis (Krauss), Monodonta turbinata (Born) and surface seawater were collected at eight locations along the coast of Ras Beirut, Lebanon, and analysed for lead, cadmium, copper, nickel and iron. With the exception of cadmium, the metal levels found in the three molluscs appear to be high in relation to levels reported by investigators from other coastal areas. The average values for lead, cadmium and nickel within these animals were quite similar. Concentrations of copper in P. coerulea were lower, but concentrations of iron were much higher than those found in both B. variabilis and M. turbinata . Further studies of metal levels in various other intertidal organisms from Ras Beirut, together with investigations into the possible effects of seasonal changes on metal concentration and distribution, are currently under way.

Mineralogical and morphological modifications through a whole annual cycle in shells of Monodonta articulata, Monodonta turbinata and Patella coerulea have been investigated and related to temperature variations. The polymorphic composition of Ca-carbonate, both in the last-formed portion and in the whole shell, appears to be correlated with temperature in M. articulata; the correlations are usually significant, except for the largest shells, at a high level of confidence and their use as a paleoecological tool is suggested. Also, the thickness of the shells is correlated with temperature, both in M. articulata and M. turbinata and such a correlation could be used in paleothermometry investigations. Patella coerulea shells do not show much sensitivity to temperature and the correlation found with aragonite percentages seems too weak to be used confidently in paleoecological investigations, while the morphological parameters are not correlated with temperature at all.

1. 1. The green pigment in the foot of Monodonta turbinata was isolated. 2. 2. The free pigment was found to be a hydrophilic substance green in acid and yellow in alkaline solution. 3. 3. The absorption spectrum of the pigment in methanolic HCl showed maxima at 680 and 370 mμ (E370/E680=0.87), and inflexions at 440, 330 and 270 mμ. The zinc complex exhibited an absorption maximum at 665–670 mνm, and in inflexion at 615 mμ, after oxidation with iodine. 4. 4. The pigment gave the Gmelin reaction. It did not react with diazotised sulphanilic acid. 5. 5. It is suggested that the pigment is a bilatriene. 6. 6. Green pigment with an identical absorption spectrum was isolated from foot tissue of the realted species Monodonta articulata.

1. 1. A blue-green pigment has been isolated from shells of the trochid Monodonta turbinata . 2. 2. The pigment behaved as a highly polar compound and gave the Gmelin reaction. 3. 3. The absorption spectrum of the pigment in methanolic HCl showed maxima at 630, 363 and 315 mμ and inflexions at 405 and 270 mμ. The zinc complex of the pigment exhibited yellow-green fluorescence in u.v. light. On oxidation with iodine it showed a shoulder at 660 mμ extending to 610 mμ in the absorption spectrum. 4. 4. It is suggested that the pigment is a biladiene probably related to the bilatriene present in the foot of M. turbinata .

THE trochids Monodonta turbinata and Gibbula divaricata are midlittoral and sublittoral animals respectively. On the rocky shores of Malta, M. turbinata is found semi-exposed in the very narrow “intertidal” zone. G. divaricata is usually found submerged in shallow water. It was considered that G. divaricata might be adapted for prolonged submersion by having among other factors a higher capacity to incur an oxygen debt than M. turbinata. The experiments described here were carried out to test this hypothesis.