Biochemical biomarkers in environmental studies—lessons learnt from enzymes catalase,glutathione <Emphasis Type="Italic">S</Emphasis>-transferase and cholinesterase in two crustacean species

Background, aim and scope  

For reliable environmental risk assessment of pollutants, knowledge on the effects at different levels of biological organisation is needed. During the early days of biomarker research in environmental studies approximately two decades ago, biochemical biomarkers were considered as the most promising tool for such purposes. Among these, three enzymes have often been studied: catalase (CAT), glutathione S-transferase (GST) and cholinesterase (ChE). However, despite their intensive research, their measurements in invertebrates have not been commonly applied in environmental risk assessment (ERA) or for regulatory purposes.     

Assessment of low-level metal contamination using the Mediterranean mussel gills as the indicator tissue

Purpose  

The aim of this study was to compare the level of metal contamination in two bays in the middle part of the Eastern Adriatic coastal zone in Croatia using the gills of mussels Mytilus galloprovincialis as indicator tissue. Despite the existing sources of contamination, previous studies with caged mussels only indicated moderate metal contamination of the Kaštela Bay, contrary to the Trogir Bay in which marina and shipyard present a probable source of Cu- and Zn-contamination.     

Photodegradation of sulfonamides and their N 4-acetylated metabolites in water by simulated sunlight irradiation: kinetics and identification of photoproducts

Once released into the aquatic environment, pharmaceuticals may undergo different degradation processes. Photodegradation, for example, might be an important elimination process for light-sensitive pharmaceuticals, such as antibiotics. In this study, the fate of sulfonamides (sulfamethazine, sulfadiazine, and sulfamethoxazole) and their N ⁴-acetylated metabolites (N ⁴-acetylsulfadiazine, N ⁴-acetylsulfamethazine, and N ⁴-acetylsulfamethoxazole) under simulated sunlight irradiation was investigated. The irradiation resulted in total or almost total degradation (88 to 98 %) of the pharmaceuticals tested, except for sulfamethazine (52 %), during 24 h of irradiation. The photoproducts of all investigated pharmaceuticals have been analyzed using high-performance liquid chromatography coupled to electrospray ionization tandem mass spectrometry. Structure elucidation performed from photodegradation products of both, sulfonamides and their N ⁴-acetylated metabolites, clearly showed two major formation pathways. These were cleavage of the sulfonamide bond as well as SO₂ extrusion. In total, nine photoproducts were elucidated. Among these photoproducts, the tautomers of sulfamethoxazole and desulfonated products of sulfadiazine and sulfamethazine were also present. Tautomers of sulfadiazine and sulfamethazine have been characterized here for the first time as well as some photoproducts of sulfadiazine, sulfametoxazole, and their metabolites N ⁴-acetylsulfadiazine and N ⁴-acetylsulfametoxazole. The obtained results are an important piece in the complex puzzle for assessing the environmental fate of sulfonamides and their metabolites in the environment.     

The Properties of Poly(<Emphasis Type="SmallCaps">l</Emphasis>-Lactide) Prepared by Different Synthesis Procedure

Different synthesis methods were applied to determine optimal conditions for polymerization of (3S)-cis-3,6-dimethyl-1,4-dioxane-2,5-dione (l-lactide), in order to obtain poly(l-lactide) (PLLA). Bulk polymerizations (in vacuum sealed vessel, high pressure reactor and in microwave field) were performed with tin(II) 2-ethylhexanoate as the initiator. Synthesis in the vacuum sealed vessel was carried out at the temperature of 150 °C. To reduce the reaction time second polymerization process was carried out in the high pressure reactor at 100 °C and at the pressure of 138 kPa. The third type of rapid synthesis was done in the microwave reactor at 100 °C, using frequency of 2.45 GHz and power of 150 W at the temperature of 100 °C. The temperature in this method was controlled via infrared system for in-bulk measuring. The solution polymerization (with trifluoromethanesulfonic acid as initiator) was possible even at the temperature of 40 °C, yielding PLLA with narrow molecular weight distribution in a very short period of time (less than 6 h). The obtained polymers had the number-average molecular weights ranging from 43,000 to 178,000 g mol⁻¹ (polydispersity index ranging from 1 to 3) according to the gel permeation chromatography measurements. The polymer structure was characterized by Fourier transform infrared and NMR spectroscopy. Thermal properties of the obtained polymers were investigated using thermogravimetry and differential scanning calorimetry.     

In vitro toxicity of selected fungicides from the groups of benzimidazoles and demethylation inhibitors to Cladobotryum dendroides and Agaricus bisporus

Twenty microfungal isolates were collected from diseased fruiting bodies of Agaricus bisporus sampled from Serbian mushroom farms during 2003–2007. Based on morphological characteristics and pathogenicity tests, the isolates were identified as Cladobotryum dendroides. The isolates of C. dendroides and A. bisporusF56 and U3 were tested for sensitivity to several selected fungicides in vitro. C. dendroides isolates were found to be more sensitive to prochloraz manganese and flusilazole + carbendazim than to the other fungicides tested (EC₅₀ values were 0.09 and 0.11 mg L^{? 1}, respectively) and weakly resistant to thiophanate-methyl (EC₅₀ values ranged between 6.53 and 12.09 mg L^{? 1}). Selectivity indexes of the tested fungicides on both C. dendroidesand A. bisporusindicated that thiophanate-methyl, cyproconazole + carbendazim and flusilazole + carbendazim had much less selective fungitoxicity than benomyl, carbendazim and prochloraz manganese.     

Pluri-annual study of the reproduction of two Mediterranean Oscarella species (Porifera, Homoscleromorpha): cycle, sex-ratio, reproductive effort and phenology

This study presents the phenology of two common Mediterranean sponges belonging to the genus Oscarella (Porifera, Homoscleromorpha). Oscarella tuberculata and Oscarella lobularis are two sibling species, dwellers of shallow benthic communities which tend to have distinct ecological behavior, respectively, euryecious and rather stenoecious. The comparative study of their reproductive cycle showed that both Oscarella species have a seasonal reproductive cycle with a successive phase duration differing from one species to another. In both species, there is a continuous oogenesis, with new oocytes appearing in spring, whereas the spermatogenesis generally starts later with the early warming of the sea. The embryonic development and the larval release are restricted to the warmest months of the year. We also observed a shift in the period of gametogenesis and larval emission depending on species and differences in their sensitivity to changes in thermal regime. It appears that an increase in seawater temperature can affect sex determination, with mainly a shift toward males in both species. Their reproductive efforts are variable in time, and can be in some cases influenced by the temperature regime. This is especially the case of O. lobularis which seems to be the most thermosensitive, its phenology responding significantly to changes in thermal regime, whereas O. tuberculata seems to be less sensitive and/or reactive. By detecting phenological changes among sponges, this study demonstrated the relevance of such monitoring to assess the possible biological response to climate change.     

Fungicide sensitivity of Trichoderma spp. from Agaricus bisporus farms in Serbia

Trichoderma species, the causal agents of green mould disease, induce great losses in Agaricus bisporus farms. Fungicides are widely used to control mushroom diseases although green mould control is encumbered with difficulties. The aims of this study were, therefore, to research in vitro toxicity of several commercial fungicides to Trichoderma isolates originating from Serbian and Bosnia-Herzegovina farms, and to evaluate the effects of pH and light on their growth. The majority of isolates demonstrated optimal growth at pH 5.0, and the rest at pH 6.0. A few isolates also grew well at pH 7. The weakest mycelial growth was noted at pH 8.0–9.0. Generally, light had an inhibitory effect on the growth of tested isolates. The isolates showed the highest susceptibility to chlorothalonil and carbendazim (ED₅₀ less than 1 mg L^?1), and were less sensitive to iprodione (ED₅₀ ranged 0.84–6.72 mg L^?1), weakly resistant to thiophanate-methyl (ED₅₀ = 3.75–24.13 mg L^?1), and resistant to trifloxystrobin (ED₅₀ = 10.25–178.23 mg L^?1). Considering the toxicity of fungicides to A. bisporus, carbendazim showed the best selective toxicity (0.02), iprodione and chlorothalonil moderate (0.16), and thiophanate-methyl the lowest (1.24), while trifloxystrobin toxicity to A. bisporus was not tested because of its inefficiency against Trichoderma isolates.     

CFD Prediction of Air Quality in the Area between Two City Vehicular Tunnels Considering Moving Cars

Concentration fields of different pollutants that spread outside two roadtunnels predicted with a CFD code will be presented. The solution domain represents the city area located between two tunnel outlets – tunnel Strahov and tunnel Mrazovka in Prague. The vicinity of both tunnels is a heavily built up area with tall buildings forming typical street canyons. The CFD modelling predicts the situation after the tunnel Mrazovka will be finished and traffic will increase considerably between both tunnels. Namely, an interest was given to the prediction of dispersion of emissions leaving both tunnel and the area touched by the traffic. For the CFD predictions, a method previously developed for moving vehicles was used. The method uses combination of Eulerian and Lagrangian approaches to moving objects and is capable of modeling different speeds and traffic rates of cars as well as traffic-induced turbulence. Influence of several meteorological parameters was studied, namely wind speed and direction and traffic parameters, like traffic rates and speed of cars. The method separates contributions from different sources to the total concentration field, namely from background, tunnel outlet and roadway. Results are presented in the form of horizontal and vertical concentration fields of NO_x.     

Anaerobic Biodegradation of Blends Based on Polyvinyl Alcohol

The presented work deals with blends composed of polyvinyl alcohol (PVA) and biopolymers (protein hydrolysate, starch, lignin). PVA does not belong to biologically inert plastics but its degradation rate (particularly under anaerobic conditions) is low. A potential solution to the issue problem lies in preparation of blends with readily degradable substrates. We studied degradation of blow-molded films made of commercial PVA and mentioned biopolymers in an aqueous anaerobic environment employing inoculation with digested activated sludge from the municipal wastewater treatment plant. Films prepared in the first experimental series were to be used for comparing biodegradation of blends modified with native or plasticized starch; in this case effect of plasticization was not proved. The degree of PVA degradation after modification with native or plasticized starch increases in a striking and practically same manner already at a starch level as low as approximately 5 wt.%. Films of the second experimental series were prepared as additionally modified with protein hydrolysate and lignin. Only lignin-modified samples exhibited a somewhat lower degree of biodegradation but regarding the measure of lignin present in blend this circumstance is not essential. Level of biodegradation with all discussed films differed only slightly—within range of experimental error.