Bioanalytical and chemical characterization of organic micropollutant mixtures in long-term exposed passive samplers from the Joint Danube Survey 4: Setting a baseline for water quality monitoring.

Monitoring methodologies reflecting the long-term quality and contamination of surface waters are needed to obtain a representative picture of pollution and identify risk drivers. This study sets a baseline for characterizing chemical pollution in the Danube River using an innovative approach, combining continuous three-months use of passive sampling technology with comprehensive chemical (747 chemicals) and bioanalytical (seven in vitro bioassays) assessment during the Joint Danube Survey (JDS4). This is one of the world's largest investigative surface-water monitoring efforts in the longest river in the European Union, which water after riverbank filtration is broadly used for drinking water production. Two types of passive samplers, silicone rubber (SR) sheets for hydrophobic compounds and AttractSPETM HLB disks for hydrophilic compounds, were deployed at nine sites for approximately 100 days. The Danube River pollution was dominated by industrial compounds in SR samplers and by industrial compounds together with pharmaceuticals and personal care products in HLB samplers. Comparison of the Estimated Environmental Concentrations with Predicted No-Effect Concentrations revealed that at the studied sites, at least one (SR) and 4-7 (HLB) compound(s) exceeded the risk quotient of 1. We also detected AhR-mediated activity, oxidative stress response, peroxisome proliferator-activated receptor gamma-mediated activity, estrogenic, androgenic, and anti-androgenic activities using in vitro bioassays. A significant portion of the AhR-mediated and estrogenic activities could be explained by detected analytes at several sites, while for the other bioassays and other sites, much of the activity remained unexplained. The effect-based trigger values for estrogenic and anti-androgenic activities were exceeded at some sites. The identified drivers of mixture in vitro effects deserve further attention in ecotoxicological and environmental pollution research. This novel approach using long-term passive sampling provides a representative benchmark of pollution and effect potentials of chemical mixtures for future water quality monitoring of the Danube River and other large water bodies.

Analysis of Mycotoxin and Secondary Metabolites in Commercial and Traditional Slovak Cheese Samples.

Cheese represents a dairy product extremely inclined to fungal growth and mycotoxin production. The growth of fungi belonging to Aspergillus, Penicillium, Fusarium, Claviceps, Alternaria, and Trichoderma genera in or on cheese leads to undesirable changes able to affect the quality of the final products. In the present investigation, a total of 68 types of commercial and traditional Slovak cheeses were analyzed to investigate the occurrence of fungal metabolites. Altogether, 13 fungal metabolites were identified and quantified. Aflatoxin M1, the only mycotoxin regulated in milk and dairy products, was not detected in any case. However, the presence of metabolites that have never been reported in cheeses, such as tryptophol at a maximum concentration level from 13.4 to 7930 µg/kg (average: 490 µg/kg), was recorded. Out of all detected metabolites, enniatin B represents the most frequently detected mycotoxin (0.06-0.71 µg/kg) in the analyzed samples. Attention is drawn to the lack of data on mycotoxins' origin from Slovak cheeses; in fact, this is the first reported investigation. Our results indicate the presence of fungal mycotoxin contamination for which maximum permissible levels are not established, highlighting the importance of monitoring the source and producers of contamination in order to protect consumers' health.

Genetic Diversity, Ochratoxin A and Fumonisin Profiles of Strains of Aspergillus Section Nigri Isolated from Dried Vine Fruits.

We investigated ochratoxin A (OTA) contamination in raisin samples purchased from Slovak markets and determined the diversity of black-spored aspergilli as potential OTA and fumonisin (FB1 and FB2) producers. The taxonomic identification was performed using sequences of the nuclear ITS1-5.8s-ITS2 region, the calmodulin and beta-tubulin genes. We obtained 239 isolates from eight fungal genera, of which 197 belonged to Aspergillus (82%) and 42 strains (18%) to other fungal genera. OTA contamination was evidenced in 75% of the samples and its level ranged from 0.8 to 10.6 µg/kg. The combination of all three markers used enabled unambiguous identification of A. carbonarius, A. luchuensis, A. niger, A. tubingensis and A. welwitschiae. The dominant coloniser, simultaneously having the highest within-species diversity isolated from our raisin samples, was A. tubingensis. Out of all analysed strains, only A. carbonarius was found to produce OTA, but in relatively high quantity (2477-4382 µg/kg). The production of FB1 and FB2 was evidenced in A. niger strains only.

Aluminium leaching from red mud by filamentous fungi.

This contribution investigates the efficient and environmentally friendly aluminium leaching from red mud (bauxite residue) by 17 species of filamentous fungi. Bioleaching experiments were examined in batch cultures with the red mud in static, 7-day cultivation. The most efficient fungal strains in aluminium bioleaching were Penicillium crustosum G-140 and Aspergillus niger G-10. The A. niger G-10 strain was capable to extract up to approximately 141 mg·L(-1) of aluminium from 0.2 g dry weight red mud. Chemical leaching with organic acids mixture, prepared according to A. niger G-10 strain's respective fungal excretion during cultivation, proved that organic acids significantly contribute to aluminium solubilization from red mud.

Penicillium strains isolated from Slovak grape berries taxonomy assessment by secondary metabolite profile.

The secondary metabolite profiles of microfungi of the genus Penicillium isolated from samples of grape berries collected in two different phases during two vegetative seasons in Slovakia is described to assess the taxonomy. Three Slovak vine regions have been selected for this study, based on their climatic differences and national economic importance. Cultures of microfungi isolated from berries were incubated on different selective media for macro and micromorphology identification. The species Penicillium brevicompactum, Penicillium crustosum, Penicillium chrysogenum, Penicillium expansum, Penicillium palitans and Penicillium polonicum were identified according to growth and morphology. The related strains were found to produce a broad spectrum of fungal metabolites, including roquefortine C, chaetoglobosin A, penitrem A, cyclopeptin, cyclopenin, viridicatin, methylviridicatin, verrucofortine, secalonic acid D, cyclopiazonic acid, fumigaclavine and mycophenolic acid. Chemotaxonomy was performed using high-performance liquid chromatography (HPLC) and mass spectrometry (MS). Dried grape berries were also analyzed allowing to assess the presence of patulin, roquefortine C and penicillic acid; this last one has been identified in dried berries but not in vitro.

Fusarium fungi and associated metabolites presence on grapes from Slovakia.

Toxinogenic Fusarium species were identified on grape berries from Slovak vineyards, and their toxic metabolites were analysed by HPLC-MS/MS. F. subglutinans, F. oxysporum, F. proliferatum, F. semitectum, F. solani, F. subglutinans, and F. verticillioides were found with varying frequency. F. oxysporum and F. proliferatum, cultured in vitro on Czapek yeast autolysate agar and yeast extract sucrose agar, produced beauvericin, in the range from 3,265 to 13,400 µg/kg, and fusaproliferin in high concentration, ranging from 49,850 to 259,500 µg/kg. A maximum value of 2.24 µg/kg has been observed for beauvericin in dried grape berries. Fumonisin B1, and fumonisin B2 were also identified, and the observed levels ranged from 500 to 2,040 µg/kg. Over 2 years (namely 2008 and 2009) many other metabolites have been identified and analysed in grape berries, in particular: avenacein Y, apicidin, aurofusarin, chlamydosporol, 2-amino-14,16-dimethyloctadecan-3-ol, enniatin A, enniatin A1, enniatin B2, enniatin B3, and equisetin.

Berries contamination by microfungi in Slovakia vineyard regions: impact of climate conditions on microfungi biodiversity.

BACKGROUND: Weather conditions can selectively promote the growth of particular fungal species, which cause rotting and spoilage of grape berries before harvest. The presence of pathogenic fungi can lead to the development of opportunist microfungi that can produce mycotoxins and cause grapes and wine contamination. OBJECTIVE: The variation of climatic conditions allowed to design a pilot study and address relevant questions for risk assessment of climate related mycotoxins production in grapes and in wine. METHODS: Microfungi contaminating berries during the vegetative period of year 2008 and 2009 in vineyards regions of Slovakia at the early veraison and at the ripening in harvest time have been identified. RESULTS: Spoilage fungi were more abundant in veraison for both years in all the studied geographical regions, with an average temperature of 20°C, humidity between 60 and 80%, and precipitation in the range 6-5mm. Much more strains of toxigenic fungi were found during veraison and ripening in the case of a year temperature variation in the range 17-22.5°C, humidity values ranging from 58 to 80% and precipitations in the range 0.7-8.4mm. CONCLUSIONS: The results suggest that there is an effect of the climatic conditions on microfungi biodiversity.