The Cocktail Effects on the Acute Cytotoxicity of Pesticides and Pharmaceuticals Frequently Detected in the Environment.

Xenobiotics never appear as single, isolated substances in the environment but instead as multi-component mixtures. However, our understanding of the ecotoxicology of mixtures is far from sufficient. In this study, three active pharmaceutical ingredients (carbamazepine, diclofenac, and ibuprofen) and three pesticides (S-metolachlor, terbuthylazine, and tebuconazole) from the most frequently detected emerging micropollutants were examined for their acute cytotoxicity, both individually and in combination, by bioluminescence inhibition in Aliivibrio fischeri (NRRL B-11177). Synergy, additive effects, and antagonism on cytotoxicity were determined using the combination index (CI) method. Additionally, PERMANOVA was performed to reveal the roles of these chemicals in binary, ternary, quaternary, quinary, and senary mixtures influencing the joint effects. Statistical analysis revealed a synergistic effect of diclofenac and carbamazepine, both individually and in combination within the mixtures. Diclofenac also exhibited synergy with S-metolachlor and when mixed with ibuprofen and S-metolachlor. S-metolachlor, whether alone or paired with ibuprofen or diclofenac, increased the toxicity at lower effective concentrations in the mixtures. Non-toxic terbuthylazine showed great toxicity-enhancing ability, especially at low concentrations. Several combinations displayed synergistic effects at environmentally relevant concentrations. The application of PERMANOVA was proven to be unique and successful in determining the roles of compounds in synergistic, additive, and antagonistic effects in mixtures at different effective concentrations.

Evaluating the In Vivo Virulence of Environmental Pseudomonas aeruginosa Using Microinjection Model of Zebrafish (Danio rerio).

(1) Background: Microinjection of zebrafish (Danio rerio) embryos offers a promising model for studying the virulence and potential environmental risks associated with Pseudomonas aeruginosa. (2) Methods: This work aimed to develop a P. aeruginosa infection model using two parallel exposition pathways on zebrafish larvae with microinjection into the yolk and the perivitelline space to simultaneously detect the invasive and cytotoxic features of the examined strains. The microinjection infection model was validated with 15 environmental and clinical strains of P. aeruginosa of various origins, antibiotic resistance profiles, genotypes and phenotypes: both exposition pathways were optimized with a series of bacterial dilutions, different drop sizes (injection volumes) and incubation periods. Besides mortality, sublethal symptoms of the treated embryos were detected and analyzed. (3) Results: According to the statistical evaluation of our results, the optimal parameters (dilution, drop size and incubation period) were determined. (4) Conclusions: The tested zebrafish embryo microinjection infection model is now ready for use to determine the in vivo virulence and ecological risk of environmental P. aeruginosa.

Survival and growth of microscopic fungi derived from tropical regions under future heat waves in the Pannonian Biogeographical Region.

Warming and heat waves are predicted by different climate models in the near future in the Pannonian Biogeographical Region (PBR). These climatic effects may have impact on the prevalence and distribution of certain fungal species of this area. In this study the effects of predicted climate scenarios were tested on fungi being endemic or unintentionally introduced by global trade from regions of warm temperate climate. Common fungal species were selected for the study and exposed to heat waves during 7 days according to two climate scenarios: one moderately (RCP 4.5, Tavg = 27 °C, Tmax = 35 °C, RH: 100%) and one strongly pessimistic (RCP 8.5, Tavg = 30 °C, Tmax = 40 °C, RH: 100%) that include predictions for the Central Hungarian Region for July 2050. According to our results, Aspergillus flavus, Aspergillus niger, Aspergillus tubingensis and Fusarium strains introduced from tropical regions tolerated heat waves, unlike Penicillium and Talaromyces spp. and endemic Cladosporium spp. which were unable to grow under the RCP 8.5 treatment. The effects of climate change on fungi raise new issues not only from economic and health perspectives, but also in relation with plant protection and environment. Our results suggest that heat waves driven by climate change promote the colonization and growth of the tested strains of non-native fungi more likely than that of the native ones.

Accumulation of HT-2 toxin from contaminated mushroom compost by edible Agaricus bisporus.

Wheat straw is commonly used as a cellulose source in mushroom compost and could be a secondary source of mycotoxin contamination in the food chain. We cultivated edible Agaricus bisporus and Pleurotus ostreatus on T-2/HT-2 artificially-contaminated mushroom compost and developed and in-house validated an UHPLC-MS/MS method for determination of T-2, HT-2, T2-triol and T2-tetraol in mushroom compost and mushroom basidiocarp. A rapid phase I metabolization of T-2 and HT-2 in mushroom compost was observed. In Agaricus bisporus, basidiocarps 8-15 µg kg-1 accumulation of HT-2 calculated on wet weight was measured. No detectable mycotoxins were found in Pleurotus ostreatus basidiocarp.

Mycological Investigation of Bottled Water Dispensers in Healthcare Facilities.

The usage of bottled water dispensers (BWDs) has spread worldwide. Despite their popularity, few studies have dealt with their microbial contaminants, and little attention is given to their fungal contamination. To our knowledge this is the first mycological study of BWDs in Europe. 36 devices have been examined in Budapest, Hungary. Despite of the strictly regulated water hygiene system in Hungary, molds and yeasts were detected in 86.8% of the samples, 56.76% were highly contaminated. Elevated heterotrophic plate counts were also observed in all samples compared to that of Hungarian drinking water. As all physical and chemical water quality characteristics have met the relevant national and European parametric values and neither totally explained the results of microbial counts, the effect of usage and maintenance habits of the devices were examined. Fungal concentrations were affected by the time elapsed since disinfection, days remaining until expiration of bottles, month of sampling and exposure to sunlight during storage. Microbes are able to proliferate in the bottled water and disperse inside the BWDs. Many of the detected fungal species (Sarocladium kiliense, Acremonium sclerotigenum/egyptiacum, Exophiala jeanselmei var. lecanii-corni, Exophiala equina, Meyerozyma guilliermondii, Cystobasidium slooffiae, Aspergillus jensenii, Bisifusarium biseptatum) are opportunistic pathogens for subpopulations of sensitive age groups and patients with immunodeficient conditions, including cystic fibrosis. Thus BWDs may pose a health risk to visitors of healthcare institutions, especially to patients with oral lesions in dental surgeries. The study draws attention to the need to investigate microbial contamination of these devices in other countries as well.