Whole Body Vibration Training Has No Effect on Vascular Endothelial and Inflammatory Markers in Young Healthy Women.

Background: The aim of the study was to comparatively assess the impact of single and repeated whole body vibration training (WBVT) and training without vibration on changes in the concentration of vascular endothelial growth factor (VEGF), endothelial nitric oxide synthase (eNOS), and high-sensitivity C-reactive protein (hsCRP) in healthy, young, non-training women. Methods: The study involved 46 women (age 20.48 ± 1.72 years), who were divided into three groups: the experimental group participating in WBVT (EVG, n = 17); the comparison group performing the same exercises but without the vibration factor (EXG, n = 12); and the control group, which did not participate in any training (CON, n = 17). The program included participation in 36 training sessions conducted over 12 weeks, with a frequency of 3 times per week. In the EVG and EXG groups, venous blood was collected before and after the first and last training sessions, while in the CON group, blood was collected twice at a 3-month interval. Results: No significant changes were observed in the concentrations of the studied markers either after a single or repeated training session in both experimental groups (p > 0.05). Conclusions: The proposed WBVT protocol appears to be a safe form of exercise that does not induce negative inflammatory reactions. The applied vibration stimulus combined with physical exercises did not initiate pro-angiogenic processes or stimulate eNOS activity in healthy women, suggesting that similar studies should be conducted in individuals with circulatory problems or chronic inflammatory diseases.

Non-interchangeable functions of efflux transporters of Pseudomonas aeruginosa in survival under infection-associated stress.

Pseudomonas aeruginosa is a challenging opportunistic pathogen due to its intrinsic and acquired mechanisms of antibiotic resistance. A large repertoire of efflux transporters actively expels antibiotics, toxins, and metabolites from cells and enables growth of P. aeruginosa in diverse environments. In this study, we analyzed the roles of representative efflux pumps from the Resistance-Nodulation-Division (RND), Major Facilitator Superfamily (MFS), and Small Multidrug Resistance (SMR) families of proteins in the susceptibility of P. aeruginosa to antibiotics and bacterial growth under stresses imposed by human hosts during bacterial infections: an elevated temperature, osmotic stress, low iron, bile salts, and acidic pH. We selected five RND pumps MexAB-OprM, MexEF-OprN, MexCD-OprJ, MuxABC-OpmB, and TriABC-OpmH that differ in their substrate specificities and expression profiles, two MFS efflux pumps PA3136-3137 and PA5158-5160 renamed here into MfsAB and MfsCD-OpmG, respectively, and an SMR efflux transporter PA1540-1541 (MdtJI). We found that the most promiscuous RND pumps such as MexEF-OprN and MexAB-OprM are integrated into diverse survival mechanisms and enable P. aeruginosa growth under various stresses. MuxABC-OpmB and TriABC-OpmH pumps with narrower substrate spectra are beneficial only in the presence of the iron chelator 2,2'-dipyridyl and bile salts, respectively. MFS pumps do not contribute to antibiotic efflux but play orthogonal roles in acidic pH, low iron, and in the presence of bile salts. In contrast, MdtJI protects against polycationic antibiotics but does not contribute to survival under stress. Thus, efflux pumps play specific, non-interchangeable functions in P. aeruginosa cell physiology and bacterial survival under stresses. IMPORTANCE: The role of multidrug efflux pumps in the intrinsic and clinical levels of antibiotic resistance in Pseudomonas aeruginosa and other gram-negative bacteria is well-established. Their functions in bacterial physiology, however, remain unclear. The P. aeruginosa genome comprises an arsenal of efflux pumps from different protein families, the substrate specificities of which are typically assessed by measuring their impact on susceptibility to antibiotics. In this study, we analyzed how deletions and overproductions of efflux pumps affect P. aeruginosa growth under human-infection-induced stresses. Our results show that the physiological functions of multidrug efflux pumps are non-redundant and essential for the survival of this important human pathogen under stress.

Effect of Whole-Body Cryotherapy on Oxidant-Antioxidant Imbalance in Women with Multiple Sclerosis.

The aim of the study was to investigate whether 20 whole-body cryotherapy treatments have an effect on oxidative-antioxidant imbalances in women with multiple sclerosis. Fifty women aged 30-55 were examined: study group-15 women with multiple sclerosis, subjected to whole-body cryotherapy; first control group-20 women with multiple sclerosis who did not receive cryotherapy intervention; second control group-15 healthy women who participated in cryotherapy treatments. Blood from the examined women was collected twice (before and after the series of 20 cryotherapy sessions). An insignificant increase in the total antioxidant capacity (study group: p = 0.706; second control group: p = 0.602) was observed after the whole-body cryotherapy intervention. After the series of cryotherapy sessions, the total oxidative status/total oxidative capacity value was insignificantly decreased among the multiple sclerosis patients (decrease by 14.03%, p = 0.495). In women with multiple sclerosis, no significant cryotherapy impact was demonstrated on changes in the oxidant-antioxidant imbalance or concentrations of nitric oxide, uric acid, or matrix metalloproteinase-9.

Respiratory Muscle Function in Older Adults with Chronic Respiratory Diseases after Pulmonary Rehabilitation in Subterranean Salt Chambers.

Training the respiratory muscles is a crucial aspect of pulmonary rehabilitation. The purpose of this study was to assess the function of respiratory muscles in older adults both before and after a period of pulmonary rehabilitation and treatment stay within the underground chambers of a salt mine. A total of 50 patients aged 65 years and older with chronic respiratory conditions was enrolled in the study. These participants underwent a 3-week subterranean pulmonary rehabilitation (PR) program in the "Wieliczka" Salt Mine. Levels of sniff nasal inspiratory pressure (SNIP), maximum inspiratory pressure (MIP), and maximum expiratory pressure (MEP) were measured using the MicroRPM both before and after the outpatient PR program conducted 135 m underground. A total of 44 patients with a mean age of 68.8 ± 2.9 years who completed the PR program and tests were included in the analysis. The average changes in the parameters of pulmonary function before and after the PR were: MIP 8.8 cmH2O, MEP 7.1 cmH2O, and SNIP 11.2 cmH2O (for p < 0.05). For patients older than 70 years, beneficial changes were only observed for MEP, which increased by 9.3 cmH2O (for p < 0.05). Speleotherapy combined with pulmonary rehabilitation improves respiratory muscle function in older adults with chronic respiratory diseases, mainly in terms of MEP. Therefore, a greater emphasis on inspiratory muscle training in the rehabilitation program should be considered.

Effect of Whole-Body Vibration Training on Hemorheological Blood Indices in Young, Healthy Women.

The aim of the study is to assess the effect of single and 12-week WBVT and training without vibration on changes in hemorheological blood indices and plasma fibrinogen levels in young, healthy women. Three groups are distinguished: the experimental group-participating in WBVT (n = 17); the comparison group-implementing the same physical exercise protocol without the vibration factor (n = 12); and the control group-no intervention (n = 17). In the experimental and comparison group, blood is collected before and after the first and last training, while in the control group, blood is collected twice, 3 months apart. After a series of WBVT, a significant decrease in the mean erythrocyte volume and mean hemoglobin mass in erythrocytes, as well as a slight increase in the mean erythrocyte hemoglobin concentration, is found, and the effect of the last training is a significant decrease in plasma volume. Under the influence of repeated WBVT, there is an increase in erythrocyte deformability at low shear stress and an increase in the aggregation amplitude. The study shows that WBVT improves blood flow in the vessels and does not affect erythrocyte aggregation and the level of fibrinogen, which confirms the safety of this form of exercise.

Functional Diversity of Gram-Negative Permeability Barriers Reflected in Antibacterial Activities and Intracellular Accumulation of Antibiotics.

Gram-negative bacteria are notoriously more resistant to antibiotics than Gram-positive bacteria, primarily due to the presence of the outer membrane and a plethora of active efflux pumps. However, the potency of antibiotics also varies dramatically between different Gram-negative pathogens, suggesting major mechanistic differences in how antibiotics penetrate permeability barriers. Two approaches are used broadly to analyze how permeability barriers affect intracellular accumulation of antibiotics. One compares the antibacterial activities of compounds, while the other measures the total intracellular concentrations of compounds in nongrowing cells, with both approaches using strains harboring wild-type or genetically modified efflux systems and permeability barriers. Whether the two assays provide similar mechanistic insights remains unclear. In this study, we analyzed the intracellular accumulation and antibacterial activities of antibiotics representative of major clinical classes in three Gram-negative pathogens of high clinical importance, Pseudomonas aeruginosa, Escherichia coli, and Acinetobacter baumannii. We found that both assays are informative about properties of permeability barriers, but there is no quantitative agreement between the assays. Our results show that the three pathogens differ dramatically in their permeability barriers, with the outer membrane playing the dominant role in E. coli and P. aeruginosa but efflux dominating in A. baumannii. However, even compounds of the same chemotype may use different permeation pathways depending on small chemical modifications. Accordingly, a classification analysis revealed limited conservation of molecular properties that define compound penetration into the three bacteria.

Acinetobacter baumannii Survival under Infection-Associated Stresses Depends on the Expression of Resistance-Nodulation-Division and Major Facilitator Superfamily Efflux Pumps.

Multidrug efflux transporters are major contributors to the antibiotic resistance of Acinetobacter baumannii in clinical settings. Previous studies showed that these transporters are tightly integrated into the physiology of A. baumannii and have diverse functions. However, for many of the efflux pumps, such functions remain poorly defined. In this study, we characterized two putative drug efflux pumps, AmfAB and AmfCD (Acinetobacter Major Facilitator), that are homologous to EmrAB-like transporters from Escherichia coli and other Gram-negative bacteria. These pumps comprise the Major Facilitator Superfamily (MFS) transporters AmfB and AmfD and the periplasmic membrane fusion proteins AmfA and AmfC, respectively. We inactivated and overproduced these pumps in the wild-type ATCC 17978 strain and its derivative strains lacking the major efflux pumps from the Resistance-Nodulation-Division (RND) superfamily and characterized antibiotic susceptibilities and growth of the strains under stresses typical during human infections. We found that neither AmfAB nor AmfCD contribute to the antibiotic non-susceptibility phenotypes of A. baumannii. The two pumps, however, are critical for the adaptation and growth of the bacterium under acidic stress, whereas AmfCD also contributes to growth under conditions of low iron, high temperature, and in the presence of bile salts. These functions are dependent on the presence of the RND pumps, the inactivation of which further diminishes A. baumannii survival and growth. Our results suggest that MFS transporters contribute to stress survival by affecting the permeability properties of the A. baumannii cell envelope.

Effect of Whole-Body Vibration on Serum Levels of Brain Derived Neurotrophic Factor and Cortisol in Young, Healthy Women.

Vibration exercises on a platform (whole-body vibration, WBV), widely used in rehabilitation, sports medicine, and fitness, is an alternative to strength effort. The presented study assessed the effect of a 12-week cycle of vibration training on the serum concentrations of brain-derived neurotrophic factor (BDNF) and cortisol in young women (trial ID: ACTRN 12621000114842). Volunteers were assigned to three groups: performing exercises on a vibrating platform (n = 17), performing identical exercises without a platform (n = 12), and passive control group (n = 17). The concentration of BDNF and cortisol was assessed four times: before the first training session, 5 min after it, also before, and 5 min after the last training session. There were no statistically significant changes in the groups or among groups for both substances. WBV in the presented form did not increase the secretion of BDNF and is not a stressful stimulus.

Dynamic Balance and Chest Mobility of Older Adults after Speleotherapy Combined with Pulmonary Rehabilitation, Endurance and Strength Training-A Prospective Study in Chronic Respiratory Diseases.

BACKGROUNDS: As people age, they are more likely to experience balance disturbances. Pulmonary rehabilitation is recognized as a core component in the management of older adults with chronic respiratory diseases. The aim of the study was to assess the dynamic balance and chest mobility of older adults participating in speleotherapy combined with pulmonary rehabilitation, endurance and strength training. METHODS: The study group consisted of 51 older adults with chronic respiratory disorders who participated in a 3-week pulmonary rehabilitation programme in underground salt chambers in the 'Wieliczka' Salt Mine Health Resort. These individuals underwent the Four Square Step Test (FSST) and circumferential chest mobility measurement before and after the outpatient rehabilitation programme conducted 135 m underground. RESULTS: Before rehabilitation in the underground salt chambers, half of the results (50%, 22 patients) were below the norm in the assessment of chest mobility between maximal inhale and exhale. The average time needed to perform FSST decreased significantly (p ≤ 0.05) from 10.2 ± 1.9 s before the stay to 9.1 ± 1.7 s after the stay and the average increase in chest mobility increased significantly (p ≤ 0.05) from 4.5 ± 5.5 cm to 5.4 ± 2.8 cm. CONCLUSIONS: Speleotherapy combined with pulmonary rehabilitation, endurance and strength training increased the dynamic balance and chest mobility of older adults with chronic respiratory diseases, as measured by the FSST and circumferential chest expansion assessment.

Making sense of drug-efflux transporters in the physiological environment.

Bacterial drug-efflux transporters act synergistically with diffusion barriers of cellular membranes and other resistance mechanisms to protect cells from antibiotics and toxic metabolites. Their critical roles in clinical antibiotic and multidrug resistance are well established. In addition, a large body of evidence has been accumulated in support of their important contributions to bacterial growth and proliferation during infections. However, how these diverse functions of drug transporters are integrated at the level of bacterial cell physiology remains unclear. This opinion briefly summarizes the current understanding of substrate specificities and physiological roles of drug-efflux pumps from Resistance-Nodulation-Division (RND) superfamily of proteins in two ESKAPE pathogens Pseudomonas aeruginosa and Acinetobacter baumannii. Based on the analysis of phenotypic and transcriptomic studies in vitro and in vivo, we propose that RND pumps of Gram-negative bacteria fall into three categories: constitutively expressed, regulated, and silent. These three categories of efflux pumps participate in different physiological programs, which are not involved in the central metabolism and bacterial growth.

Pulmonary rehabilitation in subterranean chambers combined with neuro-orthopedic activity-dependent plasticity therapy influences patients' quality of life - A preliminary study.

OBJECTIVE: The aim of the study was to evaluate if Neuro-orthopedic Activity-dependent Plasticity (N.A.P.) therapy combined with standard subterranean pulmonary rehabilitation (SPR) conducted in the salt mine influences the health-related quality of life (HRQoL) and chest mobility in patients with asthma or chronic upper respiratory tracts diseases. METHODS: 54 patients enrolled for the study completed a 3-week SPR. The 15D questionnaire, chest mobility and back scratch flexibility tests were conducted before and after SPR. In the study group (N = 23,16 women, 7 men) the N.A.P. therapy was added to SPR, while the control group (N = 31, 21 women and 10 men) completed the SPR program. RESULTS: After the SPR statistically significant improvements were observed on the 15D dimensions of breathing and vitality dimensions, in the chest mobility and back flexibility in both groups. With the baseline gender distribution, age, generic 15D score, chest mobility and back scratch adjusted, the change in the 15D score was 0.068 greater in the study group than in the control group. This difference is clinically important and statistically significant (p = 0.022). There was no statistically significant difference in the chest mobility and back scratch flexibility between the groups. CONCLUSIONS: Adding N.A.P. therapy techniques to the SPR program in the underground part of the 'Wieliczka' Salt Mine Health Resort results in a statistically significant and clinically important improvement in the subjects' HRQoL.

Effect of Whole-Body Cryotherapy on Morphological, Rheological and Biochemical Indices of Blood in People with Multiple Sclerosis.

The aim of this study was to examine and assess the impact of a series of 20 whole-body cryotherapy (WBC) treatments on the biochemical and rheological indices of blood in people with multiple sclerosis. In this prospective controlled study, the experimental group consisted of 15 women aged 34-55 (mean age, 41.53 ± 6.98 years) with diagnosed multiple sclerosis who underwent a series of whole-body cryotherapy treatments. The first control group consisted of 20 women with diagnosed multiple sclerosis. This group had no intervention in the form of whole-body cryotherapy. The second control group consisted of 15 women aged 30-49 years (mean age, 38.47 ± 6.0 years) without neurological diseases and other chronic diseases who also underwent the whole-body cryotherapy treatment. For the analysis of the blood indices, venous blood was taken twice (first, on the day of initiation of whole-body cryotherapy treatments and, second, after a series of 20 cryotherapy treatments). The blood counts were determined using an ABX MICROS 60 hematological analyzer (USA). The LORCA analyzer (Laser-Optical Rotational Cell Analyzer, RR Mechatronics, the Netherlands) was used to study the aggregation and deformability of erythrocytes. The total protein serum measurement was performed using a Cobas 6000 analyzer, Roche and a Proteinogram-Minicap Sebia analyzer. Fibrinogen determinations were made using a Bio-Ksel, Chrom-7 camera. Statistically significant differences and changes after WBC in the levels of red blood cells (RBC), hemoglobin (HGB), hematocrit (HCT), elongation index, total extend of aggregation (AMP), and proteins (including fibrinogen) were observed. However, there was no significant effect of a series of 20 WBC treatments on changes in blood counts, rheology, and biochemistry in women with multiple sclerosis. Our results show that the use of WBC has a positive effect on the rheological properties of the blood of healthy women.

Loss of RND-Type Multidrug Efflux Pumps Triggers Iron Starvation and Lipid A Modifications in Pseudomonas aeruginosa.

Transporters belonging to the resistance-nodulation-division (RND) superfamily of proteins are invariably present in the genomes of Gram-negative bacteria and are largely responsible for the intrinsic antibiotic resistance of these organisms. The numbers of genes encoding RND transporters per genome vary from 1 to 16 and correlate with the environmental versatilities of bacterial species. Pseudomonas aeruginosa strain PAO1, a ubiquitous nosocomial pathogen, possesses 12 RND pumps, which are implicated in the development of clinical multidrug resistance and known to contribute to virulence, quorum sensing, and many other physiological functions. In this study, we analyzed how P. aeruginosa's physiology adapts to a lack of RND-mediated efflux activities. A combination of transcriptomics, metabolomics, genetic, and analytical approaches showed that the P. aeruginosa PΔ6 strain, lacking the six best-characterized RND pumps, activates a specific adaptation response that involves significant changes in the abundance and activities of several transport system, quorum sensing, iron acquisition, and lipid A modification pathways. Our results demonstrate that these cells accumulate large quantities of Pseudomonas quinolone signals (PQS), which triggers iron starvation and activation of siderophore biosynthesis and acquisition pathways. The accumulation of iron in turn activates lipid A modification and membrane protection pathways. A transcriptionally regulated RND pump, MuxABC-OpmB, contributes to these transformations by controlling the concentration of coumarins. Our results suggest that these changes reduce the permeability barrier of the outer membrane and are needed to protect the cell envelope of efflux-deficient P. aeruginosa.

Defining new chemical space for drug penetration into Gram-negative bacteria.

We live in the era of antibiotic resistance, and this problem will progressively worsen if no new solutions emerge. In particular, Gram-negative pathogens present both biological and chemical challenges that hinder the discovery of new antibacterial drugs. First, these bacteria are protected from a variety of structurally diverse drugs by a low-permeability barrier composed of two membranes with distinct permeability properties, in addition to active drug efflux, making this cell envelope impermeable to most compounds. Second, chemical libraries currently used in drug discovery contain few compounds that can penetrate Gram-negative bacteria. As a result of these challenges, intensive screening campaigns have led to few successes, highlighting the need for new approaches to identify regions of chemical space that are specifically relevant to antibacterial drug discovery. Herein we provide an overview of emerging insights into this problem and outline a general approach to addressing it using prospective analysis of chemical libraries for the ability of compounds to accumulate in Gram-negative bacteria. The overall goal is to develop robust cheminformatic tools to predict Gram-negative permeation and efflux, which can then be used to guide medicinal chemistry campaigns and the design of antibacterial discovery libraries.

Inactivation of AdeABC and AdeIJK efflux pumps elicits specific nonoverlapping transcriptional and phenotypic responses in Acinetobacter baumannii.

Multidrug resistant (MDR) strains of Acinetobacter baumannii present a serious clinical challenge. The development of antibiotic resistance in this species is enabled by efflux pumps of the Resistance-Nodulation-Division (RND) superfamily of proteins creating an efficient permeability barrier for antibiotics. At least three RND pumps, AdeABC, AdeIJK, and AdeFGH are encoded in the A. baumannii genome and are reported to contribute to antibiotic resistance in clinical isolates. In this study, we analyzed the contributions of AdeABC and AdeIJK in antibiotic resistance and growth physiology of the two MDR strains, AYE and AB5075. We found that not only the two pumps have nonoverlapping substrate specificities, their inactivation leads to specific nonoverlapping changes in gene expression as determined by RNA sequencing and confirmed by gene knockouts and growth phenotypes. Our results suggest that inactivation of AdeIJK elicits broader changes in the abundances of mRNAs and this response is modified in the absence of AdeB. In contrast, inactivation of AdeB leads to a focused cellular response, which is not sensitive to the activity of AdeIJK. We identified additional efflux pumps and transcriptional regulators that contribute to MDR phenotype of clinical A. baumannii isolates.

The influence of speleotherapy combined with pulmonary rehabilitation on functional fitness in older adults - preliminary report.

OBJECTIVE: Our aim was to determine the influence of pulmonary rehabilitation conducted in therapeutic salt mine chambers on the functional fitness of older adults. METHODS: The study included 22 individuals of age >65 years with chronic respiratory conditions. The patients underwent the Fullerton test before and after a 3-week outpatient pulmonary rehabilitation in the "Wieliczka" Salt Mine Health Resort. RESULTS: After the rehabilitation stay, the results showed statistically significant improvements within five of the six parameters evaluated. In the Arm Curl, the mean number of repetitions within 30 s increased from 14.55 ± 3.63 to 16.68 ± 3.83 and in the Chair Stand from 11.86 ± 2.55 to 14.41 ± 2.95. Beneficial changes were observed in the Back Scratch, but without statistical significance. In Sit and Reach results increased from -2.3 ± 11.11cm to 2.14 ± 9.19 cm. Time for performing the 8-Foot Up and Go decreased from 6.63 ± 1.27 s to 5.8 ± 0.86 s and in 2-Minute Step results increased from 88.27 ± 20.64 to 96.55 ± 16.38 repetitions. CONCLUSION: Functional fitness of examined older adults with pulmonary disorders has increased after a rehabilitation and treatment stay in underground salt mine chambers. The reviews of this paper are available via the supplemental material section.

Factors Influencing Microbiological Biodiversity of Human Foot Skin.

The aim of the study was to analyze the microbiological biodiversity of human foot skin with respect to factors such as age, gender, frequency of foot washing and physical activity, and to select indicator species to be considered when designing textile materials with antimicrobial properties used for sock and insole production. The experiment was carried out on a group of 40 people. The number of microorganisms was determined using culture-dependent methods. Biodiversity was determined using culture followed by genetic identification based on 16S rRNA gene sequencing (bacteria), ITS region (fungi), or using Illumina next-generation sequencing (in a group of eight selected individuals). The total bacterial number on women's feet was on average 1.0 × 106 CFU/cm2, and was not statistically significantly different than that of men's feet (mean 1.2 × 105 CFU/cm2). The number of bacteria, in most cases, decreased with age and with increased frequency of physical activity. The number of bacteria increased with diminishing feet-washing frequency; however, statistically significant differences were found between groups. The number of fungi was not significantly different amongst groups. Bacteria belonging to the phyla Firmicutes, Proteobacteria and Actinobacteria constituted the main microorganisms of the foot skin. Ascomycota and Basidiomycota predominated amongst the fungi. The presence of specific species varied in groups depending on the factors mentioned above. Two of the species identified were classified as pathogens (Neisseria flavescens and Aspergillus fumigatus). These findings suggest that it is necessary to extend the list of microorganisms tested on textiles with respect to hygienic properties.

Factors Determining the Biodiversity of Halophilic Microorganisms on Historic Masonry Buildings.

The aim of the present study was to obtain insights into the relationship between the chemical (salt content and pH) and physico-mechanical (humidity and compressive strength) properties of mineral-based materials from historic buildings with salt efflorescence and the growth and biodiversity of halophilic microorganisms. Samples were mainly characterized by pH 6.5-8.5 and a moisture content of between 0.12 and 3.3%. Significant variations were also found in the salt content (sulfates, chlorides, and nitrates) of the materials. An SEM/EDS analysis of material surfaces revealed the presence of halite, calcite, gypsum, sodium sulfate, and potassium-sodium sulfate. Culture-dependent and culture-independent (clone library construction) approaches were both applied to detect halophilic microorganisms. Results derived from culturable methods and the materials analysis revealed a correlation between the total halophile count and pH value as well as sulfate content. A correlation was not observed between the concentration of chlorides or nitrates and the number of halophilic microorganisms. The materials studied were inhabited by the culturable halophilic bacteria Halobacillus sp., Virgibacillus sp., and Marinococcus sp. as well as the yeast Sterigmatomyces sp., which was isolated for the first time from mineral materials. Culture-independent techniques revealed the following bacterial species: Salinibacterium, Salinisphaera, Rubrobacter, Rubricoccus, Halomonas, Halorhodospira, Solirubrobacter, Salinicoccus, and Salinibacter. Biodiversity was the highest in materials with high or moderate salinity.

Untargeted Metabolomics Approach in Halophiles: Understanding the Biodeterioration Process of Building Materials.

The aim of the study was to explore the halophile metabolome in building materials using untargeted metabolomics which allows for broad metabolome coverage. For this reason, we used high-performance liquid chromatography interfaced to high-resolution mass spectrometry (HPLC/HRMS). As an alternative to standard microscopy techniques, we introduced pioneering Coherent Anti-stokes Raman Scattering Microscopy (CARS) to non-invasively visualize microbial cells. Brick samples saturated with salt solution (KCl, NaCl (two salinity levels), MgSO4, Mg(NO3)2), were inoculated with the mixture of preselected halophilic microorganisms, i.e., bacteria: Halobacillus styriensis, Halobacillus naozhouensis, Halobacillus hunanensis, Staphylococcus succinus, Marinococcus halophilus, Virgibacillus halodenitryficans, and yeast: Sterigmatomyces halophilus and stored at 28°C and 80% relative humidity for a year. Metabolites were extracted directly from the brick samples and measured via HPLC/HRMS in both positive and negative ion modes. Overall, untargeted metabolomics allowed for discovering the interactions of halophilic microorganisms with buildings materials which together with CARS microscopy enabled us to elucidate the biodeterioration process caused by halophiles. We observed that halophile metabolome was differently affected by different salt solutions. Furthermore, we found indications for haloadaptive strategies and degradation of brick samples due to microbial pigment production as a salt stress response. Finally, we detected changes in lipid content related to changes in the structure of phospholipid bilayers and membrane fluidity.