Microbial Diversity and Bioactive Compounds in Dried Lycium barbarum Fruits (Goji): A Comparative Study.

This study compares the microbial diversity and content of bioactive compounds in dried goji berries available on the Polish market to those of the most highly valued goji berries from the Ningxia region in China. The content of phenols, flavonoids, and carotenoids were determined, as well as the antioxidant capacities of the fruits. The quantitative and qualitative composition of the microbiota inhabiting the fruits was assessed using metagenomics by high-throughput sequencing on the Illumina platform. The highest quality was demonstrated by naturally dried fruits from the Ningxia region. These berries were characterized by a high content of polyphenols and high antioxidant activity, as well as high microbial quality. The lowest antioxidant capacity was shown by goji berries cultivated in Poland. However, they contained a high amount of carotenoids. The highest microbial contamination was found in the goji berries available in Poland (>106 CFU/g), which is important in terms of consumer safety. Despite the widely accepted benefits of consuming goji berries, both the country of cultivation and the preservation method may influence their composition, bioactivity, and microbial quality.

Bioactivities and Microbial Quality of Lycium Fruits (Goji) Extracts Derived by Various Solvents and Green Extraction Methods.

Goji berries, known for their health-promoting properties, are increasingly consumed around the world, often in the form of naturally- or freeze-dried fruits, further unprocessed. The aim of the study was to obtain dried goji berries extracts, characterized by high bioactivity and safety in terms of microbial contamination for the consumer. In the study, various solvents (water, ethanol, acetone, ethyl lactate, sunflower oil) and green extraction methods (heating and ultrasound-assisted extraction (UAE)) were used. In goji extracts, antioxidant activity and carotenoids content as bioactivity indicators, as well as total number of microorganisms were determined. Boiling of powdered dried goji fruits in water resulted in extracts with the best features, i.e., with high antioxidant properties (2.75−4.06 µmol of Trolox equivalent (TE)/mL), moderate to high content of carotenoids (0.67−1.86 mg/mL), and a reduced number of microorganisms compared with fruits. Extracts in 50% ethanol and 50% acetone were characterized primarily by very high antioxidant activity (3.09−4.90 µmol TE/mL). The high content of carotenoids (0.98−3.79 mg/mL) and high microbial quality (<10 CFU/g) were obtained by extraction in ethyl lactate by the UAE method. The results could be useful in the development of functional food based on goji berry ingredients.

Microfluidic preparation of antimicrobial microparticles composed of l-lactide/1,3-dioxolane (co)polymers loaded with quercetin.

The resistance of microorganisms against commonly used antibiotics is becoming an increasingly important problem in the food and pharmaceutical industries. Therefore, the development of novel bactericidal agents, as well as the design of drug delivery systems based on materials composed of biocompatible and biodegradable building blocks, has attracted increasing attention. To address this challenge, microparticles composed of l-lactide homopolymer and l-lactide/1,3-dioxolane (co)polymers loaded with quercetin (Q) were fabricated by using a microfluidic technique. This method enables the preparation of homogeneous particles with sizes ranging from 60 to 80 µm, composed of degradable semicrystalline or amorphous (co)polyesters. The microencapsulation of Q in a (co)polymeric matrix enables prolonged release of the antimicrobial agent. The antibacterial properties of the obtained biocompatible microparticles are confirmed by the agar diffusion plate method for various bacterial strains. Therefore, Q-loaded microparticles can have important applications in food preservation as a novel antimicrobial system.

A simple strategy for efficient preparation of networks based on poly(2-isopropenyl-2-oxazoline), poly(ethylene oxide), and selected biologically active compounds: Novel hydrogels with antibacterial properties.

Novel polymer networks composed of biocompatible, hydrophilic poly(2-isopropenyl-2-oxazoline) (PiPOx), poly(ethylene oxide) (PEO), and selected biologically active compounds (cinnamic acid, benzoic acid or eugenol) were developed for potential antimicrobial applications. The applied crosslinking method, based on the addition reaction between oxazoline pendant groups and chosen reagents containing carboxyl functions, is relatively facile, free from by-products, and thus well suited for biomaterial preparation. The one-step synthesis enabled efficient network formation with high gel contents (>90%). The chemical structure of the newly synthesized networks was characterized using Fourier Transform Infrared-attenuated Total Reflection spectroscopy (FTIR-ATR) and 13C Magic-Angle Spinning (MAS) NMR. To evaluate the suitability for biomedical applications, swelling in water and the mechanical properties of the networks were investigated. The antimicrobial efficacy of the prepared hydrogels was tested in neutral medium both by the agar diffusion method and in the liquid culture against Gram-positive and Gram-negative strains: Staphylococcus aureus, Pseudomonas aeruginosa, Escherichia coli, Klebsiella pneumoniae and Enterobacter cloaceae. All the tested hydrogels showed an antimicrobial effect in the direct contact zone. Moreover, the eugenol loaded hydrogel expressed a broader bacteriostatic effect inhibiting microorganism growth beyond the contact zone. These form-stable hydrogels with antibacterial properties may be of interest for designing materials dedicated to biomedical applications.

Antifungal Activity of Polyoxometalate-Ionic Liquids on Historical Brick.

Moulds inhabiting mineral-based materials may cause their biodeterioration, contributing to inestimable losses, especially in the case of cultural heritage objects and architectures. Fungi in mouldy buildings may also pose a threat to human health and constitute the main etiological factor in building related illnesses. In this context, research into novel compounds with antifungal activity is of high importance. The aim of this study was to evaluate the antifungal activity of polyoxometalate-ionic liquids (POM-ILs) and their use in the eradication of moulds from historical brick. In the disc diffusion assay, all the tested POM-ILs inhibited growth of a mixed culture of moulds including Engyodontium album, Cladosporium cladosporioides, Alternaria alternata and Aspergillus fumigatus. These were isolated from the surfaces of historical brick barracks at the Auschwitz II-Birkenau State Museum in Oswiecim, Poland. POM-IL coatings on historical brick samples, under model conditions, showed that two compounds demonstrated very high antifungal activity, completely limiting mould growth and development. The antifungal activity of the POM-ILs appeared to stem from their toxic effects on conidia, as evidenced by environmental scanning transmission electron microscopy observations. The results herein indicated that POM-ILs are promising disinfectant materials for use not only on historical objects, but probably also on other mineral-based materials.

Clove Oil (Syzygium aromaticum L.) Activity against Alicyclobacillus acidoterrestris Biofilm on Technical Surfaces.

Acidotermophilic bacteria Alicyclobacillus acidoterrestris is one of the main contaminants in the fruit industry forming biofilms which are difficult to remove from the production line by conventional methods. An alternative approach aims for the use of essential oils to prevent Alicyclobacillus biofilm development. The effect of clove essential oil on A. acidoterrestris biofilms on glass and polyvinyl chloride surfaces under static and agitated culture conditions was investigated by atomic force microscopy and the plate count method. The medium-flow and the type of technical surface significantly influenced A. acidoterrestris biofilm. The PVC was colonized in a greater extent comparing to glass. Clove essential oil in 0.05% (v/v) caused 25.1-65.0% reduction of biofilms on the technical surfaces along with substantial changes in their morphology by a decrease in the biofilm: height, surface roughness, and surface area difference. The oil also induced alteration in individual bacterial cells length and visible increase of their roughness. Clove essential oil seems to release EPS from biofilm and thus induce detachment of bacteria from the surface. Due to anti-A. acidoterrestris biofilm activity, the clove oil may be used in the juice industry to hinder a development of A. acidoterrestris biofilms on production surfaces.

PLA/ß-CD-based fibres loaded with quercetin as potential antibacterial dressing materials.

Microbial infections lead to elevated inflammatory responses, which usually result in prolonged and incomplete wound healing. Therefore, there is an increasing demand for biodegradable fibres that are effective against a different range of microorganisms, especially those with antibiotic resistance. Herein, quercetin-(Q)-loaded polylactide-based fibres were developed using the electrospinning technique. Since Q exhibits low chemical stability, we used star-shaped polylactides (PLAs) with a ß-CD core to host Q by inclusion complexation. To enhance the stability of the fibres and additionally entrap the Q between polymeric chains, we adapted supramolecular cross-linking by the stereocomplexation of PLAs with opposite configurations. As a control, we prepared an additional formulation of star-shaped/commercial PLA/Q for the preparation of nonwovens in which the ß-CD moiety was not present. All developed fibres were smooth and continuous, with an average diameter of 37 µm. Although nonwovens did not possess diffusible activity, good antibacterial effects against Staphylococcus aureus (S. aureus), Escherichia coli (E.coli) and Klebsiella pneumoniae (K. pneumoniae) were observed. All these features validate the proposed approach, in which different supramolecular interactions were used to modify the properties of PLA-based fibres and, most importantly, show their great potential usefulness against microbial infections.

Effect of Clove and Thyme Essential Oils on Candida Biofilm Formation and the Oil Distribution in Yeast Cells.

Candida biofilm structure is particularly difficult to eradicate, since biofilm is much more resistant to antifungal agents than planktonic cells. In this context, a more effective strategy seems to be the prevention of biofilm formation than its eradication. The aim of the study was to examine whether the process of initial colonization of materials (glass, polyethylene terephthalate, polypropylene) by food-borne Candida sp. can be impeded by clove and thyme essential oils, used at their minimal inhibitory concentrations. In the presence of clove oil, 68.4-84.2% of the yeast tested showed a statistically significant reduction in biofilm formation, depending on the material. After treatment with thyme oil, statistically significant decrease in biofilm cell numbers was observed for 63.2-73.7% of yeasts. Confocal laser scanning microscopy showed diverse compounds of clove and thyme oils that were disparately located in C. albicans cell, on a cell wall and a cell membrane, in cytoplasm, and in vacuoles, depicting the multidirectional action of essential oils. However, essential oils that were used in sub-inhibitory concentration were sequestrated in the yeast vacuoles, which indicate the activation of Candida defense mechanisms by cell detoxification. Clove and thyme essential oils due to their anti-biofilm activity can be efficiently used in the prevention of the tested abiotic surfaces colonization by Candida sp.

Multistep approach to control microbial fouling of historic building materials by aerial phototrophs.

The scientific multistep approach described herein is a result of two years of research into a control method against microbial fouling and biodeterioration of historic building materials by phototrophs. A series of tests were conducted to select the best antifouling agent for eliminating 'green' coatings and protecting surfaces against biofouling. Of the seven active compounds, two with the best penetration abilities were subjected to a photosynthetic activity inhibition test using confocal microscopy. Of the two, a quaternary ammonium salt (QAC) - didecyldimethylammonium chloride (DDAC) - was found to be the most effective. Ten biocides containing QACs at different concentrations were then tested against 'green' coatings on wood, brick and plaster, with the best four being selected for further research in model conditions. As a result, biocides containing >14% (v v-1) DDAC were found to be successful antifouling agents for protecting historical materials against biodeterioration by phototrophs.

Typing and virulence factors of food-borne Candida spp. isolates.

Food-borne yeasts, excluding yeasts used as starter cultures, are commonly considered as food spoilage microorganisms. However, the incidence of non-C. albicans Candida (NCAC) infections has increased considerably over the past two decades. Although 15 Candida species are frequently identified as pathogens, a threat to human from food-borne Candida is poorly recognized. In the present study food-borne NCAC were characterized for the virulence factors, known to be associated with yeast pathogenicity. All food-borne strains in planktonic forms and 89% in biofilm structures represented biotypes established for C. albicans, and 61% demonstrated hemolytic activity. 56-94% of food-borne isolates formed biofilms on glass and biomaterials at a level comparable to clinical C. albicans. Nine out of eighteen tested food-borne NCAC strains (C. krusei, C. lusitaniae, C. famata, C. colliculosa, C. parapsilosis, C. tropicalis) showed similarity to clinical C. albicans in terms of their biotypes and the tested virulence factors, allocating them in a group of risk of potential pathogens. However, their capacity to grow at 37 °C seems to be the preliminary criterion in the study of potential virulence of food-borne yeasts.

Exon-specific U1 snRNAs improve ELP1 exon 20 definition and rescue ELP1 protein expression in a familial dysautonomia mouse model.

Familial dysautonomia (FD) is a rare genetic disease with no treatment, caused by an intronic point mutation (c.2204+6T>C) that negatively affects the definition of exon 20 in the elongator complex protein 1 gene (ELP1 also known as IKBKAP). This substitution modifies the 5' splice site and, in combination with regulatory splicing factors, induces different levels of exon 20 skipping, in various tissues. Here, we evaluated the therapeutic potential of a novel class of U1 snRNA molecules, exon-specific U1s (ExSpeU1s), in correcting ELP1 exon 20 recognition. Lentivirus-mediated expression of ELP1-ExSpeU1 in FD fibroblasts improved ELP1 splicing and protein levels. We next focused on a transgenic mouse model that recapitulates the same tissue-specific mis-splicing seen in FD patients. Intraperitoneal delivery of ELP1-ExSpeU1s-adeno-associated virus particles successfully increased the production of full-length human ELP1 transcript and protein. This splice-switching class of molecules is the first to specifically correct the ELP1 exon 20 splicing defect. Our data provide proof of principle of ExSpeU1s-adeno-associated virus particles as a novel therapeutic strategy for FD.

Candida albicans Impairments Induced by Peppermint and Clove Oils at Sub-Inhibitory Concentrations.

Members of Candida species cause significant health problems, inducing various types of superficial and deep-seated mycoses in humans. In order to prevent from Candida sp. development, essential oils are more and more frequently applied, due to their antifungal activity, low toxicity if used appropriately, and biodegrability. The aim of the study was to characterize the early alterations in Candida albicans metabolic properties in relation to proteins and chromosomal DNA profiles, after treatment with peppermint and clove oils at sub-inhibitory concentrations. The yeasts were affected by the oils even at a concentration of 0.0075% v/v, which resulted in changes in colony morphotypes and metabolic activities. Peppermint and clove oils at concentrations ranging from 0.015× MIC (minimal inhibitory concentration) to 0.5× MIC values substantially affected the enzymatic abilities of C. albicans, and these changes were primarily associated with the loss or decrease of activity of all 9 enzymes detected in the untreated yeast. Moreover, 29% isolates showed additional activity of N-acetyl-ß-glucosaminidase and 14% isolates-α-fucosidase in comparison to the yeast grown without essential oils addition. In response to essential oils at 0.25-0.5× MIC, extensive changes in C. albicans whole-cell protein profiles were noted. However, the yeast biochemical profiles were intact with the sole exception of the isolate treated with clove oil at 0.5× MIC. The alterations were not attributed to gross chromosomal rearrangements in C. albicans karyotype. The predominantly observed decrease in protein fractions and the yeast enzymatic activity after treatment with the oils should be considered as a phenotypic response of C. albicans to the essential oils at their sub-inhibitory concentrations and may lead to the reduction of this yeast pathogenicity.

Selected Essential Oils as Antifungal Agents Against Antibiotic-Resistant Candida spp.: In Vitro Study on Clinical and Food-Borne Isolates.

Candida spp. cause significant health problems, inducing various types of superficial and deep-seated mycoses in humans. As a result of the increasing antibiotic resistance among pathogenic yeasts, the interest in alternative agents of antifungal activity is growing. This study evaluated the antimicrobial activity of selected essential oils (EOs) against Candida clinical and food-borne strains, including antibiotic-resistant isolates, in relation to yeast cell surface hydrophobicity (CSH). Candida strains showed different range of susceptibility to tea tree, thyme, peppermint, and clove oils, and peppermint oil demonstrated the lowest anticandidal activity with minimal inhibitory concentrations (MICs) of 0.03-8.0% v/v. MIC values for thyme and clove oils ranged from 0.03% to 0.25% v/v, and for tea tree oil-from 0.12% to 2.0% v/v. The exception was Candida tropicalis food-borne strain, the growth of which was inhibited after application of EOs at concentration of 8% v/v. Due to diverse yeast susceptibility to EOs, isolates were divided into five clusters in a principal component analysis model, each containing both clinical and food-borne strains. Hydrophobic properties of yeast were also diversified, and 37% of clinical and 50% of food-borne strains exhibited high hydrophobicity. The study indicates high homology of clinical and food-borne Candida isolates in relation to their susceptibility to anticandidal agents and hydrophobic properties. The susceptibility of yeasts to EOs could be partially related to their CSH. High antifungal activity of examined EOs, also against antibiotic-resistant isolates, indicates their usefulness as agents preventing the development of Candida strains of different origin.

Protection of Historical Wood against Microbial Degradation-Selection and Application of Microbiocides.

The aim of this study was to select effective and safe microbiocides for the disinfection and protection of historical wooden surfaces at the former Auschwitz II-Birkenau concentration and extermination camp. We tested seven active compounds against bacteria and moulds, of which didecyldimethylammonium chloride and N-(3-aminopropyl)-N-dodecylpropane-1,3-diamine were effective even at 0.02%-2%. Subsequently, eight microbiocides containing the selected active ingredients were chosen and applied three times on the surface of wood samples colonized by bacteria and moulds. ABM-1 and ABM-2-6% solution; Rocima 101-8%; Preventol R 80-12%; Acticide 706 LV-15% and Boramon-30% were the most effective disinfectants. Under laboratory conditions, ABM-1, Boramon and Rocima 101 ensured antimicrobial protection of new wood samples for six months. In situ, 30% Boramon and 8% Rocima 101 applied by spraying effectively protected the historical wood from bacterial and mould growth for 12 and 3 months, respectively. Colour and luminance of the new wood were not altered after exposure to the biocides. Boramon and Rocima 101, applied by the spraying method, caused no significant change in the colour of the historical wood. Results from this study were used to develop a procedure for the protection of wood in historical buildings against biodeterioration.

Proteasome machinery is instrumental in a common gain-of-function program of the p53 missense mutants in cancer.

In cancer, the tumour suppressor gene TP53 undergoes frequent missense mutations that endow mutant p53 proteins with oncogenic properties. Until now, a universal mutant p53 gain-of-function program has not been defined. By means of multi-omics: proteome, DNA interactome (chromatin immunoprecipitation followed by sequencing) and transcriptome (RNA sequencing/microarray) analyses, we identified the proteasome machinery as a common target of p53 missense mutants. The mutant p53-proteasome axis globally affects protein homeostasis, inhibiting multiple tumour-suppressive pathways, including the anti-oncogenic KSRP-microRNA pathway. In cancer cells, p53 missense mutants cooperate with Nrf2 (NFE2L2) to activate proteasome gene transcription, resulting in resistance to the proteasome inhibitor carfilzomib. Combining the mutant p53-inactivating agent APR-246 (PRIMA-1MET) with the proteasome inhibitor carfilzomib is effective in overcoming chemoresistance in triple-negative breast cancer cells, creating a therapeutic opportunity for treatment of solid tumours and metastasis with mutant p53.

Quaternary ammonium biocides as antimicrobial agents protecting historical wood and brick.

Quaternary ammonium compounds (QACs) are widely used in disinfection of water, surfaces and instruments as well as in textile, leather and food industries because of their relatively low toxicity, broad antimicrobial spectrum, non-volatility and chemical stability. Due to these advantages, QACs are also used in restoration and can be applied on historical material. The aim of this study was to determine the usefulness of biocides based on quaternary ammonium salts and containing various excipients in the protection of historical materials against microbial growth. The study determined the antimicrobial activity of three biocides against bacteria: Pseudomonas fluorescens, Staphylococcus equorum, Bacillus cereus, Bacillus muralis, Sporosarcina aquimarina and Rhodococcus fascians, and moulds: Chaetomium globosum, Penicillium citreonigrum, Cladosporium cladosporioides I, Acremonium strictum, Aspergillus fumigatus and Cladosporium cladosporioides II, all isolated from historical wood and brick. Staphylococcus equorum, Bacillus cereus, Sporosarcina aquimarina and Rhodococcus fascians bacteria, and Cladosporium cladosporioides I and Acremonium strictum moulds showed high sensitivity to quaternary ammonium biocides. Historical wood can be effectively disinfected by three applications of biocide A (30% v/v) containing dodecyl dimethyl ammonium chloride (DDAC), citric acid, propiconazole and propanol. Disinfection of historical brick can be carried out by three applications of 6% v/v solutions of biocide B (based on DDAC and ethylenediaminetetraacetic acid - EDTA) or biocide C (containing a non-ionic surfactant, DDAC and EDTA). Effective protection of historical building materials against microbial growth for a period of seven days can be achieved by the application of biocide A (30% v/v) on the wood surface and biocide B (6% v/v) on the brick surface.

Opportunistic Gram-negative rods' capability of creating biofilm structures on polivynyl chloride and styrene-acronitrile copolymer surfaces.

Biofilms are highly organized microbial communities displaying high resistance to disinfectants and other external environmental factors. Medical equipment, such as stents and catheters, can be colonized by a variety of bacteria including opportunistic pathogens circulating in the environment and dangerous to immunocompromised patients. Application of materials resistant to biofilm formation will minimize the risk of patients' infection. Hence, the aim of this research was to determine the biofilm growth of environmental bacteria isolates on polyvinyl chloride and styrene-acronitrile copolymer surfaces. Nine strains (Pseudomonas aeruginosa, Burkholderia cepacia and Serratia liquefacies) isolated from cosmetics, and a reference P. aeruginosa strain ATCC 15442, were tested. The ability and dynamics of biofilm formation on intubation catheters (30°C, up to 24 h) in bacterial growth cultures (10(7)-10(8) CFU/ml) was investigated, with subsequent sonication and quantification by agar plate count method. The results indicated that all the tested bacteria expressed a strong ability for the polymer surface adhesion, reaching 4.6 to 6.7 log CFU/cm(2) after 30 minutes. Moreover, for the majority of strains, the level of 24-hour biofilm production was from 6.67-7.61 log CFU/cm(2). This research indicates that the environmental strains circulating between the cosmetics and patients may pose a threat of biofilm formation on medical equipment surfaces, and presumably in the clinical surroundings as well.

Hydrophobic properties of Candida spp. under the influence of selected essential oils.

Processes of colonization of biotic and abiotic surfaces and biofilm formation depend inter alia on hydrophobic properties of Candida spp. The aim of this research was to determine the effect of tea tree, thyme and clove essential oils on hydrophobic properties of environmental and clinical Candida isolates. The relative cell surface hydrophobicity of strains tested was high, and ranged from 68.7% to 91.2%, with the highest value for a C. rugosa food-borne strain. The effectiveness of essential oils was diversified and depended on the type of essential oil, concentration and yeast strain. Statistically significant decrease of hydrophobicity indexes was observed after application of tea tree oil for C. krusei, clove oil for C. albicans reference strain, and all essential oils tested for C. rugosa. Only in the case of C. famata food-borne strain and C. albicans clinical isolate, solely used essential oils did not affect their hydrophobic properties. To determine the interactions of essential oils, their mixtures (1 MIC:1 MIC, 1 MIC:2 MIC and 2 MIC:1 MIC) were applied. Generally, essential oils used in combinations influenced yeast's hydrophobic properties much more than applied separately. The essential oils' mixtures reduced hydrophobicity of Candida yeasts in the range of 8.2 to 45.1%, depending on combination and strain. The interaction indexes of essential oils used in combinations predominantly indicate their additive effect. The application of tea tree, thyme and clove essential oils, especially in combinations, decreases hydrophobicity of the tested Candida isolates with implications of a probable advantageous limitation of their ability to colonize the food production industry environment.

Biodiversity of brewery yeast strains and their fermentative activities.

We investigated the genetic, biochemical, fermentative and physiological characteristics of brewery yeast strains and performed a hierarchical cluster analysis to evaluate their similarity. We used five different ale and lager yeast strains, originating from different European breweries and deposited at the National Collection of Yeast Cultures (UK). Ale and lager strains exhibited different genomic properties, but their assimilation profiles and pyruvate decarboxylase activities corresponded to their species classifications. The activity of another enzyme, succinate dehydrogenase, varied between different brewing strains. Our results confirmed that ATP and glycogen content, and the activity of the key metabolic enzymes succinate dehydrogenase and pyruvate decarboxylase, may be good general indicators of cell viability. However, the genetic properties, physiology and fermentation capacity of different brewery yeasts are unique to individual strains.

Abiotic determinants of the historical buildings biodeterioration in the former Auschwitz II-Birkenau concentration and extermination camp.

The paper presents the results of a study conducted at the Auschwitz-Birkenau State Museum in Oswiecim on the occurrence of biodeterioration. Visual assessment of the buildings revealed signs of deterioration of the buildings in the form of dampness, bulging and crumbling plaster, and wood fiber splitting. The external surfaces, and especially the concrete strips and ground immediately adjoining the buildings, were colonized by bryophytes, lichens, and algae. These organisms developed most intensively close to the ground on the northern sides of the buildings. Inside the buildings, molds and bacteria were not found to develop actively, while algae and wood-decaying fungi occurred locally. The factors conducive to biological corrosion in the studied buildings were excessive dampness of structural partitions close to the ground and a relative air humidity of above 70%, which was connected to ineffective moisture insulation. The influence of temperature was smaller, as it mostly affected the quantitative composition of the microorganisms and the qualitative composition of the algae. Also the impact of light was not very strong, but it was conducive to algae growth.