The genus Verticillium is a group of ascomycete fungi that includes several pathogenic plant species. In 2011, a new taxonomic classification, proposed by Inderbitzin and coworkers (2011), re-defined the genus as Verticillium sensu stricto. The objective of our study was the re-classification of the fungal species held in the culture collection in the Slovenian Institute of Hop Research and Brewing in accordance with the newly established taxonomy. With the PCR marker system proposed by Inderbitzin and coworkers in 2011, we re-classified 88 Verticillium isolates out of the 105 samples that are held in the institute’s bank, which were obtained from different geographic locations in Europe, North America, and Japan, and from different host plants, including alfalfa, cotton, hop, olive, potato, and tomato. However, the PCR marker for the V. dahliae identification proved to be less specific, and it resulted in the positive amplification of Gibellulopsis nigrescens, V. isaacii, and V. longisporum. To enable the accurate distinction of the fungi, the SSR and LAMP markers were added to the analyses. The 12 newly identified SSR markers, which were used in simplex PCR reactions or in combination, enabled the accurate identification of all included Verticillium isolates and could potentially be used as biomarkers for rapid and easy species identification.
Contamination with toxic metals prevents the use of sewage sludge (SS) as a soil fertilizer. Hydrodynamic cavitation, thermal microwaving, microwave-assisted alkaline, and acid hydrolysis coupled with ethylenediaminetetraacetate (EDTA) washing were tested as a method to remove toxic metals from SS. Acid hydrolysis coupled with EDTA washing was most effective and was used in a closed-loop process based on ReSoil technology. EDTA and process solutions were recycled at a pH gradient of 12.5–2, which was imposed by the addition of quicklime (CaO) and H2SO4. An average of 78%-Pb, 76%-Zn, 1%-Cu, and 17%-Cr were removed from SS in five consecutive batches. No wastewater was generated, only solid waste (40%). The EDTA lost in the process (42%) was resupplied in each batch. In a series of batches, the process solutions retained metal removal efficiency and quality. The treatment removed 70% and 23% of P and N, respectively, from SS and increased the leachability of Zn, Cu, Mn, and Fe in the washed SS by 11.7, 6.8, 1.4, and 5.2 times, respectively. Acid hydrolysis coupled with EDTA washing proved to be a technically feasible, closed-loop process but needs further development to reduce reagent, material, and nutrient loss and to reduce toxic emissions from the washed sludge.