Uncovering the Role of Autochthonous Deteriogenic Biofilm Community: Rozanec Mithraeum Monument (Slovenia).

The primary purpose of the study, as part of the planned conservation work, was to uncover all aspects of autochthonous biofilm pertaining to the formation of numerous deterioration symptoms occurring on the limestone Rozanec Mithraeum monument in Slovenia. Using state-of-the-art sequencing technologies combining mycobiome data with observations made via numerous light and spectroscopic (FTIR and Raman) microscopy analyses pointed out to epilithic lichen Gyalecta jenensis and its photobiont, carotenoid-rich Trentepohlia aurea, as the origin of salmon-hued pigmented alterations of limestone surface. Furthermore, the development of the main deterioration symptom on the monument, i.e., biopitting, was instigated by the formation of typical endolithic thalli and ascomata of representative Verrucariaceae family (Verrucaria sp.) in conjunction with the oxalic acid-mediated dissolution of limestone. The domination of lichenized fungi, as the main deterioration agents, both on the relief and surrounding limestone, was additionally supported by the high relative abundance of lichenized and symbiotroph groups in FUNGuild analysis. Obtained results not only upgraded knowledge of this frequently occurring but often overlooked group of extremophilic stone heritage deteriogens but also provided a necessary groundwork for the development of efficient biocontrol formulation applicable in situ for the preservation of similarly affected limestone monuments.

The suitability of a glovebox and of a covered still air box design for semi-sterile applications in environmental monitoring.

Laminar flow cabinets (LFCs) ensure a safe working space within which product manipulation can be carried out safely excluding contaminations of the product with the environmental microorganisms. However, for environmental monitoring applications mobile laboratories are required and these prefer the lighter gloveboxes (GB; restricted arm movement) or still air boxes (SAB; free arm movement) over the heavier, more expensive LFCs, which need to be regularly maintained. Nevertheless, the efficiency of simple GBs/SABs (no HEPA filter), in providing semi-sterile working conditions has yet to be clearly defined. Consequently, our aim was to assess the suitability of GBs/SABs for semi-sterile applications by using passive and active bioaerosol sample collection procedures within the interior spaces of these boxes. Prior to sample collection the boxes were pre-treated with different spraying preparations (70% ethanol, 2% detergent or sterile water). For a greater restriction of bioaerosol entry, SABs were constructed with covered arm ports and these were classified as partially covered (SABPC) and completely covered SABs (SABCC). Results showed that ethanol sprayed GB and SABCC exhibited microbial aerosol colony counts of zero after one hour of passive sample collection, and active sample collection revealed counts ranging between 1.9 (for GB) - 2.3 Log10CFU/m3 (for SABCC). However, ethanol sprayed SAB and SABPC were ineffective having colony counts of 6.9 and 6.5 Log10CFU/m3, respectively. Other spraying regimes resulted in even higher colony counts (up to 7.3 Log10CFU/m3). Therefore, the ethanol sprayed GB and SABCC could effectively be used for semi-sterile applications, with the SABCC allowing for an unrestricted arm movement within it.

Application of (super)cavitation for the recycling of process waters in paper producing industry.

In paper production industry, microbial contaminations of process waters are common and can cause damage to paper products and equipment as well as the occurrence of pathogens in the end products. Chlorine omission has led to the usage of costly reagents and products of lower mechanical quality. In this study, we have tested a rotation generator equipped with two sets of rotor and stator assemblies to generate developed cavitation (unsteady cloud shedding with pressure pulsations) or supercavitation (a steady cavity in chocked cavitation conditions) for the destruction of a persistent bacteria Bacillus subtilis. Our results showed that only supercavitation was effective and was further employed for the treatment of waters isolated from an enclosed water recycle system in a paper producing plant. The water quality was monitored and assessed according to the chemical (COD, redox potential and dissolved oxygen), physical (settleable solids, insolubles and colour intensity) and biological methods (yeasts, aerobic and anaerobic bacteria, bacterial spores and moulds). After one hour of treatment, a strong 4 logs reduction was achieved for the anaerobic sulphate reducing bacteria and for the yeasts; a 3 logs reduction for the aerobic bacteria; and a 1.3 logs reduction for the heat resistant bacterial spores. A 22% reduction in COD and an increase in the redox potential (37%) were observed. Sediments were reduced by 50% and the insoluble particles by 67%. For bacterial destruction in real industrial process waters, the rotation generator of supercavitation spent 4 times less electrical energy in comparison to the previously published cavitation treatments inside the Venturi constriction design.

A novel rotation generator of hydrodynamic cavitation for the fibrillation of long conifer fibers in paper production.

Refining of cellulose pulp is a critical step in obtaining high quality paper characteristics, however, this process is slow and costly especially for refining longer conifer fibers which are the preferred source for high quality paper production and give the paper its strength. In this study, we have applied a novel rotation generator of hydrodynamic cavitation for refining conifer rich pulp samples. Our results show that the device is capable of generating intense shear forces and multiple zones of developed cavitation and is successful in increasing the drainage rate of high consistency pulp (3%). The paper produced by means of the obtained pulp has higher quality because of its higher tensile index (50.5 kN m kg-1) and burst index (3 kPa m2 g-1). These physical parameters were sufficient for newsprint paper and other paper/board quality manufacture. In addition, this laboratory scale rotation generator proved to be economically efficient in comparison to the routinely employed laboratory beaters. To our knowledge, this is the first example of using hydrodynamic cavitation for the refinement of softwood fiber pulp of standard industrial consistencies (3%).

Removal of bacteria Legionella pneumophila, Escherichia coli, and Bacillus subtilis by (super)cavitation.

In sufficient concentrations, the pathogenic bacteria L. pneumophila can cause a respiratory illness that is known as the "Legionnaires" disease. Moreover, toxic Shiga strains of bacteria E. coli can cause life-threatening hemolytic-uremic syndrome. Because of the recent restrictions imposed on the usage of chlorine, outbreaks of these two bacterial species have become more common. In this study we have developed a novel rotation generator and its effectiveness against bacteria Legionella pneumophila and Escherichia coli was tested for various types of hydrodynamic cavitation (attached steady cavitation, developed unsteady cavitation and supercavitation). The results show that the supercavitation was the only effective form of cavitation. It enabled more than 3 logs reductions for both bacterial species and was also effective against a more persistent Gram positive bacteria, B. subtilis. The deactivation mechanism is at present unknown. It is proposed that when bacterial cells enter a supercavitation cavity, an immediate pressure drop occurs and this results in bursting of the cellular membrane. The new rotation generator that induced supercavitation proved to be economically and microbiologically far more effective than the classical Venturi section (super)cavitation.

Efficient inactivation of MS-2 virus in water by hydrodynamic cavitation.

The aim of this study was to accurately quantify the impact of hydrodynamic cavitation on the infectivity of bacteriophage MS2, a norovirus surrogate, and to develop a small scale reactor for testing the effect of hydrodynamic cavitation on human enteric viruses, which cannot be easily prepared in large quantities. For this purpose, 3 mL scale and 1 L scale reactors were constructed and tested. Both devices were efficient in generating hydrodynamic cavitation and in reducing the infectivity of MS2 virus. Furthermore, they reached more than 4 logs reductions of viral infectivity, thus confirming the scalability of hydrodynamic cavitation for this particular application. As for the mechanism of page inactivation, we suspect that cavitation generated OH- radicals formed an advanced oxidation process, which could have damaged the host's recognition receptors located on the surface of the bacteriophage. Additional damage could arise from the high shear forces inside the cavity. Moreover, the effectiveness of the cavitation was higher for suspensions containing low initial viral titers that are in similar concentration to the ones found in real water samples. According to this, cavitation generators could prove to be a useful tool for treating virus-contaminated wastewaters in the future.

The influence of Dekkera bruxellensis on the transcriptome of Saccharomyces cerevisiae and on the aromatic profile of synthetic wine must.

A double compartment membrane system was constructed in order to systematically study possible microbial interactions between yeasts Saccharomyces cerevisiae and Dekkera bruxellensis and their impact on wine aroma. The presence of D. bruxellensis induced 77 transcripts of S. cerevisiae. These were mostly of unknown function; however, some were involved in thiamine biosynthesis and in amino acid and polyamine transport, suggesting a competitive relationship between the two yeast species. Among the transcripts with no biological function, 14 of them were found to be the members of the PAU gene family that is associated with response to anaerobiosis stress. In separated cultures, S. cerevisiae produced glycerol which was subsequently consumed by D. bruxellensis. The concentration of ethylphenols was reduced and we assume that they were absorbed onto the surfaces of S. cerevisiae yeast walls. Also in separated cultures, D. bruxellensis formed a typical profile of aromatic esters with decreased levels of acetate esters and increased level of ethyl esters.

Monitoring of quorum-sensing molecules during minifermentation studies in wine yeast.

At high cell density or under low nutrient conditions, yeasts collectively adapt their metabolism by secreting aromatic alcohols in what is known as quorum sensing. However, the mechanisms and role of quorum sensing in yeast are poorly understood, and the methodology behind this process is not well established. This paper describes an effective approach to study quorum sensing in yeast fermentations. The separation, detection, and quantification of the putative quorum-sensing molecules 2-phenylethanol, tryptophol, and tyrosol have been optimized on a simple HPLC-based system. With the use of a phenyl HPLC column and a fluorescence detector, the sensitivity of the system was significantly increased. This allowed extraction and concentration procedures to be eliminated and the process to be scaled down to 2 mL minifermentations. Additionally, an innovative method for rapid viable-cell counting is presented. This study forms the basis for detailed studies in kinetics and regulation of quorum sensing in yeast fermentation.