The potential for bacteriophages and prophage elements in fighting and preventing the gonorrhea.

Bacteriophages are the most numerous entities on earth and are found everywhere their bacterial hosts live. As natural bacteria killers, phages are extensively investigated as a potential cure for bacterial infections. Neisseria gonorrhoeae (the gonococcus) is the etiologic agent of a sexually transmitted disease: gonorrhea. The rapid increase of resistance of N. gonorrhoeae to antibiotics urges scientists to look for alternative treatments to combat gonococcal infections. Phage therapy has not been tested as an anti-gonococcal therapy so far. To date, no lytic phage has been discovered against N. gonorrhoeae. Nevertheless, gonococcal genomes contain both dsDNA and ssDNA prophages, and viral particle induction has been documented. In this review, we consider literature data about the attempts of hunting for a bacteriophage specific for gonococci - the gonophage. We also discuss the potential application of prophage elements in the fight against N. gonorrhoeae. Temperate phages may be useful in preventing and treating gonorrhea as a scaffold for anti-gonococcal vaccine development and as a source of lytic enzymes with anti-gonococcal activity.

Evaluation of Long-Term Quality of Life Using the Foot and Ankle Outcome Score (FAOS) Questionnaire in Patients Treated by Minimally Invasive Reduction and Percutaneous Stabilization of Intra-Articular Calcaneal Fractures.

BACKGROUND Treatment of displaced intra-articular calcaneal fractures is controversial. Choosing the optimal surgical technique is very important for patient quality of life and activity. In this study, we asked the question: "What is the long-term quality of life of patients after intra-articular calcaneal fractures treated by minimally invasive reduction and percutaneous stabilization (MIRPS)?". MATERIAL AND METHODS We included 51 patients - 45 males (88%) and 6 females (12%) - who underwent MIRPS of intra-articular calcaneal fractures. The males were ages 22-63 years, with a mean age of 46 years, while the females were aged 31-63, with a mean age of 47 years. The FAOS (Foot and Ankle Outcome Score) survey was used. RESULTS Women's FAOS scores were 72-95%, with a mean of 82%, and varied according to type of fracture, surgery method used, and comorbid fractures. Men's FAOS scores were 50-95%, with a mean of 84%, and varied according to type of fracture, surgery method used, and comorbid fractures. CONCLUSIONS According to set criteria regarding the FAOS scale, mainly good and very good results were observed in patients treated surgically for intra-articular calcaneus fractures using MIRPS. Westhues' method scored a significantly higher foot rating than W-R (Westhues'-Rapala method). Patients with TTF (tongue-type fracture) scored higher in the FAOS than patients treated with JDTF (joint depression-type fracture). In other cases, no significant differences were observed.

Identification and Characterization of the First Virulent Phages, Including a Novel Jumbo Virus, Infecting Ochrobactrum spp.

The Ochrobactrum genus consists of an extensive repertoire of biotechnologically valuable bacterial strains but also opportunistic pathogens. In our previous study, a novel strain, Ochrobactrum sp. POC9, which enhances biogas production in wastewater treatment plants (WWTPs) was identified and thoroughly characterized. Despite an insightful analysis of that bacterium, its susceptibility to bacteriophages present in WWTPs has not been evaluated. Using raw sewage sample from WWTP and applying the enrichment method, two virulent phages, vB_OspM_OC and vB_OspP_OH, which infect the POC9 strain, were isolated. These are the first virulent phages infecting Ochrobactrum spp. identified so far. Both phages were subjected to thorough functional and genomic analyses, which allowed classification of the vB_OspM_OC virus as a novel jumbo phage, with a genome size of over 227 kb. This phage encodes DNA methyltransferase, which mimics the specificity of cell cycle regulated CcrM methylase, a component of the epigenetic regulatory circuits in Alphaproteobacteria. In this study, an analysis of the overall diversity of Ochrobactrum-specific (pro)phages retrieved from databases and extracted in silico from bacterial genomes was also performed. Complex genome mining allowed us to build similarity networks to compare 281 Ochrobactrum-specific viruses. Analyses of the obtained networks revealed a high diversity of Ochrobactrum phages and their dissimilarity to the viruses infecting other bacteria.

Characterization of a Unique Bordetella bronchiseptica vB_BbrP_BB8 Bacteriophage and Its Application as an Antibacterial Agent.

Bordetella bronchiseptica, an emerging zoonotic pathogen, infects a broad range of mammalian hosts. B. bronchiseptica-associated atrophic rhinitis incurs substantial losses to the pig breeding industry. The true burden of human disease caused by B. bronchiseptica is unknown, but it has been postulated that some hypervirulent B. bronchiseptica isolates may be responsible for undiagnosed respiratory infections in humans. B. bronchiseptica was shown to acquire antibiotic resistance genes from other bacterial genera, especially Escherichia coli. Here, we present a new B. bronchiseptica lytic bacteriophage-vB_BbrP_BB8-of the Podoviridae family, which offers a safe alternative to antibiotic treatment of B. bronchiseptica infections. We explored the phage at the level of genome, physiology, morphology, and infection kinetics. Its therapeutic potential was investigated in biofilms and in an in vivo Galleria mellonella model, both of which mimic the natural environment of infection. The BB8 is a unique phage with a genome structure resembling that of T7-like phages. Its latent period is 75 ± 5 min and its burst size is 88 ± 10 phages. The BB8 infection causes complete lysis of B. bronchiseptica cultures irrespective of the MOI used. The phage efficiently removes bacterial biofilm and prevents the lethality induced by B. bronchiseptica in G. mellonella honeycomb moth larvae.

Antibiotics Act with vB_AbaP_AGC01 Phage against Acinetobacter baumannii in Human Heat-Inactivated Plasma Blood and Galleria mellonella Models.

Increasing multidrug resistance has led to renewed interest in phage-based therapy. A combination of the bacteriophages and antibiotics presents a promising approach enhancing the phage therapy effectiveness. First, phage candidates for therapy should be deeply characterized. Here we characterize the bacteriophage vB_AbaP_AGC01 that poses antibacterial activity against clinical Acinetobacter baumannii strains. Moreover, besides genomic and phenotypic analysis our study aims to analyze phage-antibiotic combination effectiveness with the use of ex vivo and in vivo models. The phage AGC01 efficiently adsorbs to A. baumannii cells and possesses a bacteriolytic lifecycle resulting in high production of progeny phages (317 ± 20 PFU × cell-1). The broad host range (50.27%, 93 out of 185 strains) against A. baumannii isolates and the inability of AGC01 to infect other bacterial species show its high specificity. Genomic analysis revealed a high similarity of the AGC01 genome sequence with that of the Friunavirus genus from a subfamily of Autographivirinae. The AGC01 is able to significantly reduce the A. baumannii cell count in a human heat-inactivated plasma blood model (HIP-B), both alone and in combination with antibiotics (gentamicin (GEN), ciprofloxacin (CIP), and meropenem (MER)). The synergistic action was observed when a combination of phage treatment with CIP or MER was used. The antimicrobial activity of AGC01 and phage-antibiotic combinations was confirmed using an in vivo larva model. This study shows the greatest increase in survival of G. mellonella larvae when the combination of phage (MOI = 1) and MER was used, which increased larval survival from 35% to 77%. Hence, AGC01 represents a novel candidate for phage therapy. Additionally, our study suggests that phages and antibiotics can act synergistically for greater antimicrobial effect when used as combination therapy.

Bacteriophages as Factories for Eu2O3 Nanoparticle Synthesis.

The use of phage display to identify peptides with an ability to bind and synthesize Eu2O3 nanoparticles is demonstrated in this report. This is the first report of modified phages specifically binding a lanthanide. The peptides exposed on virions revealed very strong binding to Eu2O3 nanoparticles and the ability to catalyze Eu2O3 nanoparticles' formation from Eu(OH)3 and Eu(NO3)3 solutions. The luminescence emission spectrum of Eu3+ ions indicated that these ions existed mostly in sites deviated from the inversion symmetry in crystalline Eu2O3 aggregates and gelatinous Eu(OH)3 precipitate. The ability of phage-displayed peptides to catalyze formation of Eu2O3 nanoparticles provides a useful tool for a low-cost and effective synthesis of lanthanide nanoparticles, which serve as attractive biomedical sensors or fluorescent labels, among their other applications.

Radiological results and clinical complications after calcaneal articular fracture surgical treatment using minimally invasive percutaneous fixation.

The choice of method for surgical treatment of articular calcaneal fractures remains problematic due to the potential complications and difficulty in restoring loss of functional status, anatomical geometry and congruence of joint surfaces. The preferred method of surgical treatment for these injuries, among others, is still minimally invasive percutaneous fixation. The aim of this study was to evaluate the radiological results of articular calcaneal fractures surgical treatment using a minimally invasive percutaneous fixation and to determine the prevalence of complications in early and long-term observation. Radiological results of surgical treatment of 82 patients operated using minimally invasive percutaneous fixation were analyzed. The radiological results were based on Bohler angle and varus angle using criteria according to Harnroongroj and Golec. Postoperative complications were registered in a 3 months follow-up (early observation) and after 6 months (long-term observation). Patients with tongue-type fractures had significantly better surgical outcomes than patients with depression-type fractures. Displacements of bone fragments in articular calcaneal fractures visualized radiographically by the changes of Harnroongroj et al. angle do not constitute a serious surgical problem and are easily repositioned and stabilized, as opposed to displacements resulting in changes in the value of the Böhler angle. The most common postoperative complications were local inflammatory reactions at the point of entry of fixation material and thromboembolic complications. Results of surgical treatment of articular fractures of the calcaneus using a minimally invasive percutaneous fixation are good and satisfactory in majority of patients suggesting that this method constitutes proper choice of treatment.

Phage-Directed Synthesis of Photoluminescent Zinc Oxide Nanoparticles under Benign Conditions.

Biological systems, especially bacteriophages and peptides, are an attractive green alternative to other known methods of nanoparticle synthesis. In this work, for the first time, bacteriophages were employed to synthesize a specific peptide, capable of producing nanoparticles (NPs). Derivatives of M13 bacteriophage exposing a ZnO-binding peptide (TMGANLGLKWPV) on either pIII or pVIII phage coat protein were constructed and used as a biotemplate. The exposition of the ZnO-binding peptide, synthesized by phages during their propagation in bacteria, on M13 virions provided a groundwork for growing ZnO nanostructures. Depending on the recombinant phage type used (M13-pIII-ZnO or M13-pVIII-ZnO), well separated ZnO NPs or complex 3D structures of ZnO NPs of ca. 20-40 nm were synthesized at room temperature. The synthesized ZnO nanoparticles served as a luminescent material that emitted light near the short wavelength end of the visible region (at ca. 400 nm). The next very low intensity emission band at 530 nm demonstrated that the ZnO material obtained is characterized by a low concentration of surface defects.

Modified Filamentous Bacteriophage as a Scaffold for Carbon Nanofiber.

With the advent of nanotechnology, carbon nanomaterials such as carbon nanofibers (CNF) have aroused substantial interest in various research fields, including energy storage and sensing. Further improvement of their properties might be achieved via the application of viral particles such as bacteriophages. In this report, we present a filamentous M13 bacteriophage with a point mutation in gene VII (pVII-mutant-M13) that selectively binds to the carbon nanofibers to form 3D structures. The phage-display technique was utilized for the selection of the pVII-mutant-M13 phage from the phage display peptide library. The properties of this phage make it a prospective candidate for a scaffold material for CNFs. The results for binding of CNF by mutant phage were compared with those for maternal bacteriophage (pVII-M13). The efficiency of binding between pVII-mutant-M13 and CNF is about 2 orders of magnitude higher compared to that of the pVII-M13. Binding affinity between pVII-mutant-M13 and CNF was also characterized using atomic force microscopy, scanning electron microscopy, and transmission electron microscopy, which confirmed the specificity of the interaction of the phage pVII-mutant-M13 and the CNF; the binding occurs via the phage's ending, where the mutated pVII protein is located. No similar behavior has been observed for other carbon nanomaterials such as graphite, reduced graphene oxide, single-walled carbon nanotubes, and multiwalled carbon nanotubes. Infrared spectra confirmed differences in the interaction with CNF between the pVII-mutant-M13 and the pVII-M13. Basing on conducted research, we hypothesize that the interactions are noncovalent in nature, with π-π interactions playing the dominant role. Herein, the new bioconjugate material is introduced.

Main health-related quality-of-life issues in patients after a distal radius fracture.

AIM: The aim of this study was to identify and assess the degree and clinical course of the main health-related quality-of-life (HRQoL) issues in patients after a distal radius fracture (DRF). MATERIALS AND METHODS: Patients were eligible if they were between 18-80 years and were within 1-3 days after a non-comminuted DRF. All patients filled out the Polish version of the IOF QLQ, the SF-36 and a demographic questionnaire. Assessment points were set as soon as possible after the fracture, 7 days, 6 weeks, 3 months, and 6 after the fracture. Standard statistical analyses were performed. RESULTS: During the 16 month recruitment period a total of 71 patients (55 women - 77.5%), with a mean age of 64.1 ± 12.4 years, were included in the study group. All patients suffered from Colles type fractures. Attrition to follow-up was acceptable. At baseline, basing on the IOF QLQ scale scores, DRF patients had the most significant problems with physical function (82.8/100; with 100 representing the worst possible HRQoL), and general health (78.1/100). Basing on SF-36 scale scores patients most significant problems were associated with role limitations due to physical health problems (15.1/100; with 100 representing the best possible HRQoL), and bodily pain (39.5/100). CONCLUSIONS: Concluding, this study shows that the main issues with which patients with and extra-articular DRF struggle the most are pain of the fractured extremity and physical dysfunction. These symptoms are most pronounced in the early post-injury period, and in the majority of patients steadily decrease over a period of six months.

"Pros and cons" of total hip arthroplasty with metaphyseal Proxima endoprosthesis.

The authors present their own experience concerning total hip arthroplasty with the metaphyseal prosthesis Proxima. Proxima, a metaphyseal prosthesis, provides an innovative supplement to total hip arthroplasty. In this study, the authors present their own experience using Proxima in patients with hip osteoarthritis. This study was performed between 2008 to 2013 and comprised of 62 patients, of which 38 were male (61.3%) and 24 female (38.7%). All patients were operated on due to hip osteoarthritis using total hip arthroplasty with the metaphyseal prosthesis Proxima. The age of patients included into the study ranged from 23 years to 62 years with the mean age of 46 years. The authors paid close attention to the "pros and cons" of surgical techniques and assessed clinical and radiological results in both the short and long-term observation periods. The clinical evaluation was based on the Harris Hip Score and radiological assessment of fixation of the acetabular cup of the endoprosthesis was based on Pradhan's criteria. The endoprosthesis Proxima stem was positioned using the manufacturer's recommended method, evaluating the direction, scope and duration of the displacement in the marrow cavity of the proximal stump of the femur. According to the authors' analysis, the surgery gives good functional and radiological results both in the short- and long-term observation periods. The effectiveness depends on precise qualification for surgery, proper surgical techniques and specific anatomical conditions of the proximal femur stump. The most common reasons for primary and secondary dislocations of the metaphyseal endoprosthesis Proxima stem occur during the first three months post surgery. This is due to incorrect surgical techniques, which disregard the importance of specific anatomical conditions of the proximal femoral stump, which affects Proxima implantation, and cause deviations towards a varus or valgus orientation.

Assessment of the Relationship between Antero-Posterior Dental Malocclusions, Body Posture Abnormalities and Selected Static Foot Parameters in Adults.

Objectives: This study aimed to find if a relationship exists between antero-posterior malocclusions and the level of musculoskeletal disorders in adults, including body posture and static foot analysis. Methods: In all, 420 participants were recruited through convenience sampling (Kraków University students and patients of a local dentist's practice). Following general medical interviews, dental examinations and consideration of inclusion and exclusion criteria, 90 healthy volunteers (ages 19-35) were enrolled and assigned to three groups (n = 30) based on occlusion type (Angle's molar Class I, II or III). The research procedure involved occlusion and temporomandibular disorder assessment conducted by a dental specialist. Comprehensive morphological measurements of body asymmetry were performed using the Videography 2D package and FreeSTEP software, which calculated the parameters determined from anterior, posterior and lateral projection photos. Foot loading distribution was analyzed using the FreeMED baropodometric platform. Results: Significant differences were demonstrated in the positioning of the head, cervical and lumbar spine in the sagittal plane among individuals with the analyzed occlusal classes (p < 0.05). Individuals with Angle's Class II exhibited significantly greater forward head positions and greater depths of cervical and lumbar lordosis compared with individuals with Class III or Class I. Those with overbites had higher forefoot loading. The Class III individuals exhibited greater L-R displacement, indicating a larger angle of displacement of the centers of the right and left feet relative to the lower edge of the measurement platform, suggesting pelvic rotation. Conclusions: An inclination for concurrent occurrences of malocclusions and posture deviations in the sagittal plane was observed. An interdisciplinary approach involving dentistry and physiotherapy specialists which utilizes tools for comprehensive posture assessment is crucial for diagnosing and treating such conditions.

An effective antibiofilm strategy based on bacteriophages armed with silver nanoparticles.

The emerging antibiotic resistance in pathogenic bacteria is a key problem in modern medicine that has led to a search for novel therapeutic strategies. A potential approach for managing such bacteria involves the use of their natural killers, namely lytic bacteriophages. Another effective method involves the use of metal nanoparticles with antimicrobial properties. However, the use of lytic phages armed with nanoparticles as an effective antimicrobial strategy, particularly with respect to biofilms, remains unexplored. Here, we show that T7 phages armed with silver nanoparticles exhibit greater efficacy in terms of controlling bacterial biofilm, compared with phages or nanoparticles alone. We initially identified a novel silver nanoparticle-binding peptide, then constructed T7 phages that successfully displayed the peptide on the outer surface of the viral head. These recombinant, AgNP-binding phages could effectively eradicate bacterial biofilm, even when used at low concentrations. Additionally, when used at concentrations that could eradicate bacterial biofilm, T7 phages armed with silver nanoparticles were not toxic to eukaryotic cells. Our results show that the novel combination of lytic phages with phage-bound silver nanoparticles is an effective, synergistic and safe strategy for the treatment of bacterial biofilms.

Proteomic profiles and kinetics of development of bacteriophage T4 and its rI and rIII mutants in slowly growing Escherichia coli.

Bacteriophage T4 survival in its natural environment requires adjustment of phage development to the slow bacterial growth rate or the initiation of mechanisms of pseudolysogeny or lysis inhibition (LIN). While phage-encoded RI and probably RIII proteins seem to be crucial players in pseudolysogeny and LIN phenomena, the identity of proteins involved in the regulation of T4 development in slowly growing bacteria has remained unknown. In this work, using a chemostat system, we studied the development of wild-type T4 (T4wt) and its rI (T4rI) and rIII (T4rIII) mutants in slowly growing bacteria, where T4 did not initiate LIN or pseudolysogeny. We determined eclipse periods, phage propagation times, latent periods and burst sizes of T4wt, T4rI and T4rIII. We also compared intracellular proteomes of slowly growing Escherichia coli infected with either T4wt or the mutants. Using two-dimensional PAGE analyses we found 18 differentially expressed proteins from lysates of infected cells. Proteins whose amounts were different in cells harbouring T4wt and the mutants are involved in processes of replication, phage-host interactions or they constitute virion components. Our data indicate that functional RI and RIII proteins - apart from their already known roles in LIN and pseudolysogeny - are also necessary for the regulation of phage T4 development in slowly growing bacteria. This regulation may be more complicated than previously anticipated, with many factors influencing T4 development in its natural habitat.

Is There a Correlation between Dental Occlusion, Postural Stability and Selected Gait Parameters in Adults?

Background: There is still an ongoing debate about the role of the craniomandibular system, including occlusal conditions, on postural stability. This study aims to assess the role of antero-posterior malocclusion on postural control and plantar pressure distribution during standing and walking. Methods: 90 healthy volunteers (aged 19 to 35) were qualified for the study. The subjects were assigned to three groups, depending on the occlusion type. Each group (Angle Class I, II and III) consisted of 30 people. The research procedure included a clinical occlusal assessment performed by a dentist. Postural control measurements were carried out using a force platform by measuring plantar pressure distribution during standing (six trials with and without visual control) and walking test conditions. Results: The tendency to shift the CoP forward is demonstrated by Angle Class II subjects and backwards by Class I and III subjects (p < 0.001). Individuals with a malocclusion demonstrated significantly higher selected stabilographic parameters while standing on both feet (with eyes open and closed) and during the single-leg test with eyes open (p < 0.05). The analysis of the dynamic test results showed no significant correlations between Angle Classes and the selected gait parameters. Conclusions: Analyses conducted among individuals with malocclusions showed the impact of occlusion on static postural stability. In order to diagnose and effectively treat malocclusion, a multidisciplinary approach with the participation of dentists and physiotherapy specialists is necessary, with the use of stabilometric and kinematic posture assessments.

The effect of sulfur baths on hemorheological properties of blood in patients with osteoarthritis.

Balneotherapy is an effective treatment method in various diseases and commonly used treatment modality among patients with musculoskeletal disorders. Sulfur baths are known for healing properties however effect on rheological properties is unstudied. Thus the aim of our study was to determine the effect of sulfur balneotherapy on hemorheological blood indices. A total of 48 patients with osteoarthritis were enrolled to the study. Blood samples were collected twice, before and after 3-week time period. We evaluated complete blood count, fibrinogen, hs-CRP and blood rheology parameters such as elongation index (EI), half-time of total aggregation (T1/2) and aggregation index (AI) analyzed with the Lorrca Maxis. Mean age of studied cohort was 67 ± 5 years. After sulfur baths WBC count was significantly decreased is studied group (p = 0.021) as well as neutrophile count (p = 0.036). Red blood cell EIs were statistically higher after sulfur baths in shear stress ranging from 8.24 to 60.30 Pa. T1/2 was significantly higher (p = 0.031) and AI lower (p = 0.003) compared to baseline. No significant changes in fibrinogen and hs-CRP were observed. It is the first study that evaluate effect of sulfur balneotherapy on rheologic properties of blood. Sulfur water baths may improve erythrocyte deformability and aggregation parameters.

Bovine Serum Albumin - Hydroxyapatite Nanoflowers as Potential Local Drug Delivery System of Ciprofloxacin.

Introduction: Hybrid nanoflowers are structures consisting of organic (enzymes, proteins, nucleic acids) and inorganic components (mostly metal phosphates) with a flower-like hierarchical structure. Novel hybrid nanoflowers based on bovine serum albumin (BSA) and hydroxyapatite (HA) were obtained and characterized. Study on BSA-HA nanoflowers as potential drug delivery system is reported for the first time. Methods: Embedding ciprofloxacin in the structure of hybrid nanoflowers was confirmed by ATR-FTIR and thermogravimetric analysis. The inorganic phase of the nanoflowers was determined by X-ray diffraction. UV‒Vis spectroscopy was used to evaluate the release profiles of ciprofloxacin from nanoflowers in buffer solutions at pH 7.4 and 5. The agar disk diffusion method was used to study the antibacterial activity of the synthesized nanoflowers against Staphylococcus aureus and Pseudomonas aeruginosa. Results: Bovine serum albumin - hydroxyapatite nanoflowers were obtained with diameters of ca. 1-2 µm. The kinetics of ciprofloxacin release from nanoflowers were described by the Korsmeyer-Peppas model. The antibacterial activity of the synthesized nanoflowers was demonstrated against S. aureus and P. aeruginosa, two main pathogens found in osteomyelitis. Conclusion: The formulated nanoflowers may act as an efficient local antibiotic delivery system. Due to the use of nonhazardous, biodegradable components and benign synthesis, hybrid nanoflowers are very promising drug delivery systems that could be applied in the treatment of skeletal system infections.

Revealing the diversity of bacteria and fungi in the active layer of permafrost at Spitsbergen island (Arctic) - Combining classical microbiology and metabarcoding for ecological and bioprospecting exploration.

Arctic soils are constantly subjected to extreme environmental conditions such as low humidity, strong winds, high salinity, freeze-thaw cycles, UV exposition, and low nutrient availability, therefore, they have developed unique microbial ecosystems. These environments provide excellent opportunities to study microbial ecology and evolution within pristine (i.e. with limited anthropogenic influence) regions since the High Arctic is still considered one of the wildest and least explored environments on the planet. This environment is also of interest for the screening and recovery of unique microbial strains suitable for various biotechnological applications. In this study, a combination of culture-depended and culture-independent approaches was used to determine the cultivation bias in studies of the diversity of cold-active microorganisms. Cultivation bias is a reduction in recovered diversity, introduced when applying a classical culturing technique. Six different soil types, collected in the vicinity of the Polish Polar Station Hornsund (Spitsbergen, Norway), were tested. It was revealed that the used media allowed recovery of only 6.37 % of bacterial and 20 % of fungal genera when compared with a culture-independent approach. Moreover, it was shown that a combination of R2A and Marine Broth media recovered as much as 93.6 % of all cultivable bacterial genera detected in this study. Based on these results, a novel protocol for genome-guided bioprospecting, combining a culture-dependent approach, metabarcoding, next-generation sequencing, and genomic data reuse was developed. With this methodology, 14 psychrotolerant, multi-metal-resistant strains, including the highly promising Rhodococcus spp., were obtained. These strains, besides increased metal tolerance, have a petroleum hydrocarbon utilization capacity, and thus may be good candidates for future bioremediation technologies, also suited to permanently cold regions.

Inhibition of development of Shiga toxin-converting bacteriophages by either treatment with citrate or amino acid starvation.

OBJECTIVES: Shiga toxin-producing Escherichia coli (STEC) are pathogenic strains, whose virulence depends on induction of Shiga toxin-converting prophages and their subsequent lytic development. We explored which factors or conditions could inhibit development of these phages, potentially decreasing virulence of STEC. MATERIALS AND METHODS: Lytic development of Shiga toxin-converting bacteriophages was monitored after mitomycin C-provoked prophage induction under various conditions. Phage DNA replication efficiency was assessed by measurement of DNA amount in cells using quantitative polymerase chain reaction. RESULTS: We demonstrated that the use of citrate delayed Shiga toxin-converting phage development after prophage induction. This effect was independent on efficiency of prophage induction and phage DNA replication. However, an excess of glucose reversed the effect of citrate. Amino acid starvation prevented the phage development in bacteria both able and unable to induce the stringent response. CONCLUSIONS: Lytic development of Shiga toxin-converting bacteriophages can be inhibited by either the presence of citrate or amino acid starvation. We suggest that the inhibition caused by the latter condition may be due to a block in prophage induction or phage DNA replication or both. APPLICATIONS: Our findings may facilitate development of procedures for treatment of STEC-infected patients.

Into the Unknown: Microbial Communities in Caves, Their Role, and Potential Use.

Caves have been an item of amateur and professional exploration for many years. Research on the karst caves has revealed great diversity of bacteria, algae, and fungi living on stone walls and speleothems, in mud puddles or sediments. They have become the source of interest for various research groups including geologists, chemists, ecologists, or microbiologists. The adaptations of cave-dwelling organisms applied to their survival are complex and some of their properties show potential to be used in various areas of human life. Secondary metabolites produced by cave's bacteria show strong antimicrobial, anti-inflammatory, or anticancer properties. Furthermore, bacteria that can induce mineral precipitation could be used in the construction industry and for neutralization of radioisotopes. In this review we focus on bacteria and algae present in cave ecosystems, their role in shaping such specific environment, and their biotechnological and medical potential.