Mihael J. Toman

The evaluation of waste, surface and ground water quality using the Allium test procedure

The bulbs of Allium cepa were grown in test liquids of various pollution levels as follows: undiluted industrial and municipal waste water; biological treatment plant output water; water from the Drava river upstream and downstream of the city of Maribor; and non-chlorinated drinking water as a negative control test. The paper presents the response of the Allium cepa genetic material to the presence of potential cytotoxic and genotoxic substances in test liquids and the suitability of the Allium cepa testing procedure as a method for short-term determination of water pollution level. The suitability of the Allium test procedure as a system for environmental monitoring is presented. The influence of water pollution on macroscopic and cytologic parameters of the common onion by application of the biological testing method was examined. The macroscopic parameter was inhibition of root growth. The cytological parameters were: aberrant cells in metaphase and anaphase, index of micronuclei appearance and inhibition of cell division. The possibility of categorization the different polluted test liquids into quality classes is presented according to the influence of the test liquids on macroscopic and cytologic parameters. Test liquids are divided into 8 quality classes: the first class is the least polluted surface waters, the second and the third classes are more polluted surface water, the fourth and the fifth classes are biological treatment plant output waters, the sixth till the eighth quality classes are untreated waste waters. The most polluted test liquids (untreated industrial and municipal waste waters) caused sublethal and even lethal effects. The most polluted tested liquids cause the inhibition of root growth over 50% (even up to 74%), decrease of mitotic index over 36% (even up to 66%), increase of presence of interphase cells with micronuclei over 3% and increase of presence of aberrant cells for more than 10 times in comparison to control test.

Periphyton as a bioindicator of mercury pollution in a temperate torrential river ecosystem

Mercury presents a potential risk to the environment and humans, especially in its methylated form. It is among the highest priority environmental pollutants. River Idrijca (Slovenia) is highly contaminated with mercury due to past mercury mining. The aim of this work was to investigate whether the periphyton community in rivers such as Idrijca is a suitable indicator of Hg pollution and of changes in mercury methylation and could serve as an early warning system of increased input of MeHg in the food chain. Periphyton is the only site of primary production in temperate torrential rivers such as Idrijca and is therefore an important link in the food chain. It is also a potential site of Hg accumulation and its introduction to higher trophic levels. Our aim was to assess the response of the periphyton to seasonal and spatial variations in mercury levels and to evaluate its potential as an early warning system of changes in mercury reactivity and mobilization The results indicate that periphyton in a torrential river is too complex and unpredictable to be used as a sole indicator of mercury concentrations and changes in the river. Nevertheless, it can complement environmental measurements due to its importance in the riverine food web.

TRACE ELEMENT CONCENTRATIONS IN THE TISSUES OF PROTEUS ANGUINUS (AMPHIBIA, CAUDATA) AND THE SURROUNDING ENVIRONMENT

The concentrations of some essential (Cu, Zn, Se) and some toxicelements (Hg, As) were determined in tissues (liver, kidneys, integument, and muscle) of the endemic cave salamander, Proteus anguinus Laurenti 1768 and Proteus anguinus parkelj ssp.n. captured in the Planina Cave, Kompolje Cave, and Jelševnik, all situated within the Dinaric Karst in Slovenia. The highest amounts of selenium and mercury were found in the liver of Proteus anguinus specimens from all three localities. Some of the animals from the Planina Cave showed higher concentrations of copper in the liver and muscle. Pigmented subspecies of Proteus from Jelševnik showedincreased values of arsenic in the integument and of zinc in theliver and integument. The liver of the animals contained the highest amounts of the metals analysed and therefore may be considered as a target organ. Metal levels were also measured inthe cave water and sediments. The waters were not polluted withmetals. However, the metals were relatively high in the sediments, but not to levels considered contaminated. In additionto our study of metal concentrations in Proteus tissues andhabitats, several water quality parameters were measured in the water of each locality. The main differences between the threelocalities concerned dissolved oxygen concentration and saturation, while all other physical and chemical parameterswere characteristic of the aquatic environment in the calcareousKarst region.

Efficiency of advanced oxidation processes in lowering bisphenol A toxicity and oestrogenic activity in aqueous samples

Summary Bisphenol A (BPA) is a well-known endocrine disruptor with adverse oestrogen-like effects eliciting adverse effects in humans and wildlife. For this reason it is necessary to set up an efficient removal of BPA from wastewaters, before they are discharged into surface waters. The aim of this study was to compare the efficiency of BPA removal from aqueous samples with photolytic, photocatalytic, and UV/H2O2 oxidation. BPA solutions were illuminated with different bulbs (halogen; 17 W UV, 254 nm; and 150 W UV, 365 nm) with or without the TiO2 P-25 catalyst or H2O2 (to accelerate degradation). Acute toxicity and oestrogenic activity of treated samples were determined using luminescent bacteria (Vibrio fischeri), water fleas (Daphnia magna), zebrafish embryos (Danio rerio), and Yeast Estrogen Screen (YES) assay with genetically modified yeast Saccharomyces cerevisiae. The results confirmed that BPA is toxic and oestrogenically active. Chemical analysis showed a reduction of BPA levels after photolytic treatment and 100 % conversion of BPA by photocatalytic and UV/H2O2 oxidation. The toxicity and oestrogenic activity of BPA were largely reduced in photolytically treated samples. Photocatalytic oxidation, however, either did not reduce BPA toxic and oestrogenic effects or even increased them in comparison with the baseline, untreated BPA solution. Our findings suggest that chemical analysis is not sufficient to determine the efficiency of advanced oxidation processes in removing pollutants from water and needs to be complemented with biological tests. Izvleček A je dobro poznan motilec endokrinega sistema z estrogensko aktivnostjo in škodljivimi učinki na ljudi in živali. Zaradi tega se pojavlja potreba po učinkovitem odstranjevanju BPA iz odpadnih voda. V raziskavi smo proučevali učinkovitost odstranjevanja BPA iz vodnih vzorcev s procesi fotolitske, fotokatalitske in UV/H2O2 oksidacije. Med poskusom smo vodne vzorce osvetljevali s tremi različnimi žarnicami (halogensko, 17 W UV, 254 nm in 150 W UV, 365 nm). V procesih foto(kata)litske oksidacije smo uporabili H2O2 in katalizator TiO2 P-25 (Degussa). Akutno strupenost in estrogensko aktivnost BPA ter tretiranih vzorcev smo določili s testi strupenosti na luminiscentne bakterije (Vibrio fischeri), vodne bolhe (Daphnia magna), zarodke cebric (Danio rerio) in YES testom z gensko spremenjenimi kvasovkami Saccharomyces cerevisiae. Rezultati so pokazali, da je BPA za vodne organizme strupen in estrogensko aktiven. Kemijske analize so pokazale zmanjšanje vsebnosti BPA po fotolitski oksidaciji in popolno odstranitev BPA iz vodnih vzorcev po fotokatalitski oksidaciji in UV/H2O oksidaciji. Strupenost in estrogenska aktivnost sta se zmanjšali pri vzorcih, tretiranih s fotolitsko oksidacijo, sorazmerno z zmanjšanjem vsebnosti BPA. Pri fotokatalitsko tretiranih vzorcih se kljub popolni pretvorbi BPA strupenost in estrogenska aktivnost nista zmanjšali ali pa sta se celo zvišali v primerjavi z izhodnim vzorcem. Ugotovili smo, da ugotavljanje učinkovitosti odstranjevanja BPA iz vodnih vzorcev z naprednimi oksidacijskimi procesi z uporabo kemijskih analiz ne zadostuje. Potrebna so biološka testiranja, ki dokažejo morebitno preostalo strupenost in estrogensko aktivnost obdelanih vzorcev.

Adverse effects of bisphenol A on water louse (Asellus aquaticus).

Experiments were performed to study the effects of short and long-term exposure to bisphenol A (BPA) on a freshwater crustacean isopod Asellus aquaticus (L.). Two life stages of isopods were exposed to a range of BPA concentrations, from aqueous and two dietary sources, in the form of with BPA spiked conditioned alder leaf (Alnus glutinosa) discs, or spiked formulated sediment, to determine the relative importance of each source of exposure on the uptake of this contaminant. Several lethal and sublethal endpoints were evaluated in this study to measure the potential effects of BPA on A. aquaticus, including mortality, growth and feeding rate inhibition, mobility inhibition, de-pigmentation and molting disturbances. They signify a correlation to BPA levels and a difference in BPA uptake efficiency from different uptake sources. Results of acute exposure to BPA show a greater sensitivity of test systems using juvenile specimens with a 96 h LC₅₀ of 8.6 mg L(-1) BPA in water medium and a 96 h LC₅₀ of 13.5 mg L(-1) BPA in sediment. In comparison, adult isopods show a 96 h LC₅₀ of 25.1 mg L(-1) BPA in water medium and a 96 h LC₅₀ of 65.1 mg L(-1) BPA in sediment. Observed endpoints of chronic exposures suggest the alder leave discs to be the most efficient uptake source of BPA, in contrast to uptake from water or heterogeneous sediment. Significant (p<0.05) growth inhibition, with a 21d NOEC of 0.5/2.5 mg L(-1) (for juvenile/adult organisms), and feeding rate inhibition, with a 21d NOEC of 0.5/1.0 mg L(-1) (for juvenile/adult organisms), were proven to be the most sensitive toxicity endpoints. An even more sensitive effect turned out to be molting frequency, which was significantly reduced; a 21d NOEC was 1.0 mg L(-1) of BPA for adult organisms and an even lower 21d NOEC of 0.05 mg L(-1) of BPA for juveniles. The observed endpoints are recorded at very low, non-toxic exposure concentrations, indicating that BPA acts as an endocrine disrupting compound, as well as a toxic substance. We also determined the importance of the direct dietary uptake of the pollutants, significant for juveniles as well as adult animals.