Induction and reversion of multixenobiotic resistance in the marine snail Monodonta turbinata

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.     

Metabolic fate of aromatic amines in the mussel Mytilus galloprovincialis

The postmitochondrial fraction of the digestive gland from the marine mussel Mytilus galloprovincialis possesses FAD-containing monooxygenase (EC 1.14.38) but lacks cytochrome P-450 dependent benzo(a)pyrene monooxygenase (EC 11.4.14.1). This is also evidenced by the ability of the mussel preparation to activate carcinogenic aromatic amines, but not carcinogenic benzo(a)pyrene, to Salmonella typhimurium TA 98 mutagens. This metabolic activity is NADPH dependent. Mussel digestive gland postmitochondrial fraction also possesses the enzymes needed for the detoxicating part of the aromatic amine metabolism: UDP-glucuronyl transferase (EC 2.4.1.17) and -glucuronidase (EC 3.2.1.31). Under the experimental conditions used here, this aromatic amine metabolic pathway converts up to 8% of 2-acetylamino(9-¹⁴C)fluorene, but not (G-³H)benzo(a)pyrene, to water soluble glucuronides. Glucuronic acid stimulates the formation of these glucuronides. The metabolites liberated from these glucuronides by the -glucuronidase treatment could be converted to TA 98 strain mutagens by the carp liver postmitochondrial fraction, but not by the mussel's digestive gland preparation. The presence of such a selective potential for the bioactivation and detoxication of aromatic amines, and not polycyclic aromatic hydrocarbons, in the marine invertebrate(s) may bring new insight to our understanding of the effects and the fate of carcinogens in the marine environment.     

Fostering collaboration between universities regarding regional sustainability initiatives – the University of Maribor

This paper presents the current position and role of the University of Maribor (UM) in linking stakeholders, in order to achieve sustainable regional development. A strategic approach is proposed for facilitating collaboration of the UM with regional development agencies, the local community and industry, based on collaborative learning and development. Furthermore, elements contributing to a successful learning partnership are identified together with the barriers. The results show that regional stakeholders (university, municipality, NGOs, and companies) are commonly engaged in a sustainable regional development, but their activities are not sufficiently linked. The strategic approach, and collaborative learning have led to many on-going SD projects within the University of Maribor. Room for improvement exists, regarding external university collaboration, especially in the fields of research, knowledge transfer, technological development, dissemination of information, education, and innovations.     

Density-dependent effects control the reproductive strategy and population growth of <Emphasis Type="Italic">Aurelia</Emphasis><Emphasis Type="Italic">aurita</Emphasis> s.l. scyphistomae

Aureliaaurita s.l. scyphistomae are capable of developing different asexual modes for propagation and thus present a multi-mode reproductive strategy. The reproduction rates and the reproductive strategy they adopt depend on a combination of various environmental parameters. We investigated the A.aurita s.l. polyp-to-polyp reproduction strategy and population growth in relation to polyp density. Our results confirmed that density-dependent factors control population growth of A. aurita s.l. scyphistomae in three different ways: (1) decreasing the polyp reproduction rate, (2) triggering the production of motile bud-like tissue particles and (3) inducing the detachment of developed scyphistomae. Whereas the decrease in the reproduction rate reduces the number of recruits, the motile particles and the detachment of scyphistomae contribute to minimizing density-dependent effects by allowing reproductive products and scyphistomae to drift away. Thus, not only are the negative effects of intraspecific competition for space and food diminished but also the potential colonization of new substrates, and further increase in scyphistoma density is favoured on larger spatial scales. The potential capability to switch its polyp-to-polyp reproduction strategy in response to environmental clues and population density may give Aurelia high adaptability in the temperate coastal waters where they commonly live and where they may be exposed to wide-ranging and fluctuating environmental variables that affect their survival and longevity. Considering these features, as well as the wide tolerance of Aurelia scyphistomae (and medusae) to environmental parameters, it is not surprising that the species/lineages of Aurelia are cosmopolitan and exhibit the most frequent bloom events worldwide.     

Changes in soil characteristics during landfill leachate irrigation of Populus deltoides

The effects of wastewater application on electrical conductivity, water retention and water repellency of soils planted with Populus deltoides (eastern cottonwood) and irrigated with different concentrations of landfill leachate and compost wastewater, tap water and nutrient solution were evaluated. Substrate water content at field capacity (?0.033 MPa) and at permanent wilting point (?1.5 MPa) was determined with a pressure plate extractor to assess available water capacity of the substrate. A water drop penetration test was used to determine substrate water repellency. The biomass of nutrient and landfill leachate treatments was significantly (P < 0.05) greater compared to the tap water and compost wastewater treatments. All treatments increased substrate water content at field capacity and at permanent wilting point. Landfill leachate significantly increased available water capacity (up to 52%); treatment with compost wastewater significantly decreased it (25–47%). All substrates showed increased water repellency after the experiment at field capacity and permanent wilting point comparing to the original substrate. The strongest influence on water repellency at both field capacity and permanent wilting point showed irrigation with compost wastewater and tap water. Pronounced influence on substrate’s water repellency of compost wastewater could be contributed to a high content of dissolved organic carbon, whereas Mg and Ca cations caused flocculation and consequent water repellency of the substrate irrigated with tap water. The results indicate that soil physical characteristics must be closely monitored when landfill leachate and compost wastewater are used for irrigation to avoid long term detrimental effects on the soil, and consequently on the environment. Due to the complexity of the compost wastewater quality the latter should be applied on open fields only after prior pre-treatment to reduce dissolved organic carbons, or alternatively, compost wastewater should be added only intermittently and in diluted ratios.     

Ecotoxicity of wastes in avoidance tests with Enchytraeus albidus,Enchytraeus crypticus and Eisenia fetida (Oligochaeta)

Contact bioassays are important for testing the ecotoxicity of solid materials. However, survival and reproduction tests are often not practical due to their duration which may last for several weeks. Avoidance tests with soil invertebrates may offer an alternative or extension to the classic test batteries due to their short duration (days rather than weeks) and due to a sensitive sub-acute endpoint (behavior). The aims of our study were: (a) to evaluate the effects of three solid industrial wastes (incineration ash, contaminated wood chips and contaminated soil) on three Oligochaeta species (enchytraeids Enchytraeus albidus, Enchytraeus crypticus and earthworm Eisenia fetida) in avoidance tests; (b) to compare the sensitivity among the species and to compare results of avoidance test to reproduction tests; (c) to elucidate if measuring the weight in the earthworm avoidance test could be reasonable additional endpoint. Avoidance mostly increased with the increasing percent of waste in the mixture showing a dose–response curve. E. fetida was the most sensitive species and E. crypticus the least one. An additional endpoint, (changes in weight after two-day exposure) was not found to be more sensitive than avoidance reaction, but it confirmed that earthworms staying in the highest concentrations of the waste mixture were affected showing apparent weight reduction. Our results indicate that avoidance tests with earthworms and enchytraeids are feasible for waste testing.     

Mixed sex allocation strategies in a polytocous mammal,the house mouse (<Emphasis Type="Italic">Mus musculus</Emphasis>)

The issue of adaptive adjustment of offspring sex ratio (proportion of male births) in polytocous mammals, producing several offspring per litter, is controversial because females of these species can maximize their fitness mainly by adjusting offspring number. To address this issue, we examined the effect of maternal condition at mating, experimentally decreased by pre-mating food restriction, on the sex ratio variation in 137 female mice. We tested two basic sex allocation hypotheses plausible for polytocous mammals: (1) the Myers hypothesis, predicting that cheaper sex should be favored in poor environmental conditions to maximize offspring number; and (2) the Williams hypothesis, predicting maximum fitness returns by adjusting size- and sex-specific composition of the litter according to the maternal condition. The food-restricted mothers produced larger litters with a higher proportion of cheaper daughters than the control mothers. By contrast, the control mothers optimized size and sex composition of the litter according to their weight at mating. In addition, the offspring of the food-restricted mothers suffered less from pre-weaning mortality than those of the control mothers. Therefore, when comparing the groups, the Myers hypothesis had a general significance while the Williams hypothesis was plausible only for the control mothers. Furthermore, some of the food-restricted mothers partly coped with the pre-mating food restriction and increased the proportion of sons in the litter with the increasing maternal weight loss (during the period of food restriction). The sex ratio variation was thus a result of three sex allocation strategies depending on the maternal condition at mating.