A total of 209 samples of various cereal crops (maize, wheat, barley, rye and oats) grown in Croatian fields during 2016 and 2017 were collected to analyze and determine the occurrence and co-occurrence of EU regulated mycotoxins in cereals (AFB1, AFB2, AFG1, AFG2, DON, FB1, FB2, ZEA, T-2, HT-2 and OTA). The analysis, performed by a validated confirmatory LC-MS/MS method based on a dilute and shoot principle, highlighted Fusarium mycotoxins as the main contaminants, often co-occurring in samples from both years (50.0% in 2016 and 33.7% in 2017). DON was found to be the most frequent mycotoxin, present in 72.5% of the 2016 samples and 32.6% of the 2017 samples, while maize proved to be the most contaminated cereal type of both years with FUM as the most abundant mycotoxins, with an average concentration of 1180 µg/kg. Moderate temperatures with periods of high humidity favored the accumulation of DON in wheat samples instead of other Fusarium mycotoxins, while similar conditions favored maize contamination with FUM. A total of 8.3% of all the 2016 harvest samples and 7.9% of the 2017 harvest samples were assessed as non-compliant, containing mycotoxins in concentrations higher than the levels set by the EU legislation for food.

The availability of reliable sensitive multi-analyte methods for unambiguous determination of mycotoxins is crucial for ensuring food and feed safety, considering their adverse health effects and (co-)occurrence in various foods. Accordingly, a multi-mycotoxin confirmatory method for simultaneous determination of 11 mycotoxins regulated in cereals within the European Union (EU) using ultra-high performance liquid chromatography coupled to tandem mass spectrometry (UHPLC-MS/MS) was developed and in-house validated to fit the EU legislation requirements for analytical methods. A simple sample preparation was based on a solid–liquid extraction using a solvent mixture acetonitrile/water/formic acid (79/20/1, v/v/v) and a dilution of raw extract using water/acetonitrile/formic acid (79/20/1, v/v/v) before instrumental analysis. Average recoveries in all three validated cereal crop types (maize, wheat, and barley), spiked at multiple levels, were found acceptable for all analytes when matrix-matched calibration was used, ranging from 63.2% to 111.2% and also showing very good repeatability, with relative standard deviations below 20%. Matrix effect (SSE) evaluation revealed maize as the most complex of the three analyzed cereal matrices, with strong SSE (150%) recorded for all 11 analyzed mycotoxins. An additional method verification was performed through successful participation in proficiency testing schemes, with the achieved z-scores generally in the acceptable range of −2 ≤ z ≤ 2. The obtained validation results demonstrated the suitability of the developed confirmatory multi-mycotoxin UHPLC-MS/MS method based on a dilute-and-shoot principle for the simultaneous determination of low concentrations of 11 EU-regulated mycotoxins in cereals, including aflatoxins B1, B2, G1 and G2, deoxynivalenol, fumonisins B1 and B2, zearalenone, T-2 and HT-2 toxins, and ochratoxin A.

Anaerobic digestion, despite its preferable use as a treatment for high organic matter polluted waste streams, is susceptible to inhibitors, salt included. Therefore, two different experiments were conducted to observe the responses of bacterial and archaeal communities to hypersaline environments. In the first experiment, salt was added gradually, while in the second experiment, salt was added rapidly (so-called salt shocks were performed). The results of the gradual addition of salt showed a recovery of methane production after the salt concentration decreased. The NaCl concentration of 28.2 g/L seems to be the limit between stable operation and occurrence inhibition. The specific biogas production varied between 0.490 and 0.562 m³/kg_tCOD during the stepwise salt addition, depending on the salt concentration, while the maximal achieved COD removal was 79.8%. The results of the rapid salt addition showed good recovery of the bacterial community, while a reduction of salt-sensitive species was observed in the archaeal community. The trend of specific biogas production during rapid salt addition was stable with an average value of 0.590 m³/kg_tCOD, and it was observed that higher concentrations of up to 39.4 g/L of NaCl were tolerated. The maximum COD removal achieved during rapid salt addition was 83.1%. In conclusion, certain bacterial and archaeal communities were well-adapted to the hypersaline environment and remained active during the anaerobic digestion of substrates with high salt concentration.

Biopolymers are a suitable alternative for the ongoing problem of plastic accumulation, even though commercialization is difficult, which is reflected in the price of the product. However, costs can be reduced if active sludge bacteria and cheap, accessible substrates such as waste brewer’s yeast are used. Waste brewer’s yeast is a rich source of carbon and nitrogen and is widespread as a substrate in various industries. Thus, the cultivation of active sludge bacteria was performed on waste brewers’ yeast to obtain biopolymers that can be used in 3D printing. FT-IR, TG, and DSC analyses of produced polymers were conducted after extraction, as well as biogas and biomethane potential tests. Results of cultivation under various conditions show that biopolymer content is extremely heterogeneous. However, during cultivation in SBR, signals at 1741.3, 1709.6, 1634.3, and 1238 cm⁻¹ were detected. Further analyses are needed, but when said results are compared to those of consulted scientific articles, there is an indication that at least a small amount of PHA is present in biomass produced in SBR. Biopolymers produced in SBR were used as a material for the 3D printing of a cube. Moreover, testing of the physical properties (Young’s modulus) of a 3D-printed cube was performed. After conducting experiments, it can be concluded that said process, although time-consuming, achieved the goal of printing a stable and rigid 3D-printed cube made from biopolymers. Further optimization of said process should focus on more detailed microbial selection as well as biopolymer extraction. In that way, isolation, purification, and identification techniques will be improved, which could achieve higher biopolymer yield and, thus, make biopolymers more accessible in various industries.