Hyaluron-Based Bionanocomposites of Silver Nanoparticles with Graphene Oxide as Effective Growth Inhibitors of Wound-Derived Bacteria.

Keeping wounds clean in small animals is a big challenge, which is why they often become infected, creating a risk of transmission to animal owners. Therefore, it is crucial to search for new biocompatible materials that have the potential to be used in smart wound dressings with both wound healing and bacteriostatic properties to prevent infection. In our previous work, we obtained innovative hyaluronate matrix-based bionanocomposites containing nanosilver and nanosilver/graphene oxide (Hyal/Ag and Hyal/Ag/GO). This study aimed to thoroughly examine the bacteriostatic properties of foils containing the previously developed bionanocomposites. The bacteriostatic activity was assessed in vitro on 88 Gram-positive (n = 51) and Gram-negative (n = 37) bacteria isolated from wounds of small animals and whose antimicrobial resistance patterns and resistance mechanisms were examined in an earlier study. Here, 69.32% of bacterial growth was inhibited by Hyal/Ag and 81.82% by Hyal/Ag/GO. The bionanocomposites appeared more effective against Gram-negative bacteria (growth inhibition of 75.68% and 89.19% by Hyal/Ag and Hyal/Ag/Go, respectively). The effectiveness of Hyal/Ag/GO against Gram-positive bacteria was also high (inhibition of 80.39% of strains), while Hyal/Ag inhibited the growth of 64.71% of Gram-positive bacteria. The effectiveness of Hyal/Ag and Hyal/Ag/Go varied depending on bacterial genus and species. Proteus (Gram-negative) and Enterococcus (Gram-positive) appeared to be the least susceptible to the bionanocomposites. Hyal/Ag most effectively inhibited the growth of non-pathogenic Gram-positive Sporosarcina luteola and Gram-negative Acinetobacter. Hyal/Ag/GO was most effective against Gram-positive Streptococcus and Gram-negative Moraxella osloensis. The Hyal/Ag/GO bionanocomposites proved to be very promising new antibacterial, biocompatible materials that could be used in the production of bioactive wound dressings.

Wounds of Companion Animals as a Habitat of Antibiotic-Resistant Bacteria That Are Potentially Harmful to Humans-Phenotypic, Proteomic and Molecular Detection.

Skin wounds and their infections by antibiotic-resistant bacteria (ARB) are very common in small animals, posing the risk of acquiring ARB by pet owners or antibiotic resistance gene (ARG) transfer to the owners' microbiota. The aim of this study was to identify the most common pathogens infecting wounds of companion animals, assess their antibiotic resistance, and determine the ARGs using culture-based, molecular, and proteomic methods. A total of 136 bacterial strains were isolated from wound swabs. Their species was identified using chromogenic media, followed by MALDI-TOF spectrometry. Antibiotic resistance was tested using disc diffusion, and twelve ARGs were detected using PCRs. The dominant species included Staphylococcus pseudintermedius (9.56%), E. coli, and E. faecalis (both n = 11, 8.09%). Enterobacterales were mostly resistant to amoxicillin/clavulanic acid (68.3% strains), all Pseudomonas were resistant to ceftazidime, piperacillin/tazobactam, imipenem, and tylosin, Acinetobacter were mostly resistant to tylosin (55.5%), all Enterococcus were resistant to imipenem, and 39.2% of Staphylococci were resistant to clindamycin. Among ARGs, strA (streptomycin resistance), sul3 (sulfonamide resistance), and blaTEM, an extended-spectrum beta-lactamase determinant, were the most frequent. The risk of ARB and ARG transfer between animals and humans causes the need to search for new antimicrobial therapies in future veterinary medicine.

In Vitro Antibacterial Activity of Ozonated Olive Oil against Bacteria of Various Antimicrobial Resistance Profiles Isolated from Wounds of Companion Animals.

Frequent colonization and bacterial infection of skin wounds in small animals prevent or impair their healing. However, the broadly applied antimicrobial therapy of wounds is not always necessary and promotes the spread of bacterial resistance. Thus, alternatives to antimicrobial therapy, including preventive measures in the form of wound dressings with antibiotic properties, should be searched for. The aim of this study was to develop a new, efficient, cost-effective and non-toxic formulation with antimicrobial properties to serve as an alternative to antibiotic administration in wound-healing stimulation in companion animals. Nano/microencapsulated ozonated olive oil in a hyaluronan matrix was developed, with ozone concentration high enough to prevent bacterial growth. The presence and size of nano- and microcapsules were determined with scanning electron microscopy (SEM). Antibacterial activity of developed formulations was examined in vitro on 101 Gram-positive and Gram-negative bacteria isolated from the wounds of companion animals. The highest ozone concentration in the developed formulations inhibited the growth of 40.59% bacteria. Species and genus-specific differences in reactions were observed. Enterococcus spp. proved the least susceptible while non-pathogenic Gram-positive bacteria were the most susceptible to the examined formulations. Changes in the bacterial morphology and cell structure of Psychrobacter sanguinis suspension mixed with Ca-stabilized formulations with nano/microencapsulated ozonized olive oil were revealed during SEM observations. The combination of compounds that promote wound healing (hyaluronic acid, olive oil, ozone and calcium) with the antibacterial activity of the developed formula makes it a promising bionanocomposite for use as a topical dressing.

The Preparation of Silver and Gold Nanoparticles in Hyaluronic Acid and the Influence of Low-Pressure Plasma Treatment on Their Physicochemical and Microbiological Properties.

Nanometals constitute a rapidly growing area of research within nanotechnology. Nanosilver and nanogold exhibit significant antimicrobial, antifungal, antiviral, anti-inflammatory, anti-angiogenic, and anticancer properties. The size and shape of nanoparticles are critical for determining their antimicrobial activity. In this study, silver and gold nanoparticles were synthesized within a hyaluronic acid matrix utilizing distilled water and distilled water treated with low-pressure, low-temperature glow plasma in an environment of air and argon. Electron microscopy, UV-Vis and FTIR spectra, water, and mechanical measurements were conducted to investigate the properties of nanometallic composites. This study also examined their microbiological properties. This study demonstrated that the properties of the composites differed depending on the preparation conditions, encompassing physicochemical and microbiological properties. The application of plasma-treated water under both air and argon had a significant effect on the size and distribution of nanometals. Silver nanoparticles were obtained between the range of 5 to 25 nm, while gold nanoparticles varied between 10 to 35 nm. The results indicate that the conditions under which silver and gold nanoparticles are produced have a significant effect on their mechanical and antibacterial properties.

Preparation of Nano/Microcapsules of Ozonated Olive Oil in Hyaluronan Matrix and Analysis of Physicochemical and Microbiological (Biological) Properties of the Obtained Biocomposite.

Hydrogels, based on natural polymers, such as hyaluronic acid, are gaining an increasing popularity because of their biological activity. The antibacterial effect of ozone is widely known and used, but the instability the gas causes, severely limits its application. Ozone entrapment in olive oil by its reaction with an unsaturated bond, allows for the formation of stable, therapeutically active ozone derivatives. In this study, we obtained an innovative hydrogel, based on hyaluronic acid containing micro/nanocapsules of ozonated olive oil. By combination of the biocompatible polymer with a high regenerative capacity and biologically active ingredients, we obtained a hydrogel with regenerative properties and a very weak inhibitory effect against both bacterial commensal skin microbiota and pathogenic Candida-like yeasts. We assessed the stability and rheological properties of the gel, determined the morphology of the composite, using scanning electron microscopy (SEM) and particle size by the dynamic light scattering (DLS) method. We also performed Attenuated total reflectance Fourier transform infrared (FTIR-ATR) spectroscopy. The functional properties, including the antimicrobial potential were assessed by the microbiological analysis and in vitro testing on the HaCat human keratinocyte cell line. The studies proved that the obtained emulsions were rheologically stable, exhibited an antimicrobial effect and did not show cytotoxicity in the HaCat keratinocyte model.

How much of antibiotics can enter surface water with treated wastewater and how it affects the resistance of waterborne bacteria: A case study of the Bialka river sewage treatment plant.

This study aimed to gain insight into the presence of antibiotics, occurrence of antimicrobial resistance and prevalence of extended-spectrum ß-lactamase (ESBL) genes in Escherichia coli in surface water, based on the example of the Bialka river, located in one of the most attractive tourist destinations in Poland. Water samples were collected in three sites: in the Tatra National Park (TNP), by the sewage discharge from the local treatment plant (STP) and c.a. 3 km downstream (DSTP). The analyses included determination of antibiotic content, enumeration of bacterial indicators of poor water quality, isolation and identification of Escherichia coli, which was subjected to antimicrobial susceptibility tests and assessment of ESBL-determining genes. Fourteen antimicrobials out of 24 tested were detected in river waters in varying concentrations. Trimethoprim and ofloxacin were most frequently detected. Most antibiotics were absent in the TNP, the highest numbers and the highest concentrations of antibiotics were observed by the STP discharge to decrease their content downstream. Culture-based tests of microbiological contamination showed similar results. Resistance to ampicillin was most frequent (64.5% strains), followed by cefazolin (50%). 20.6% of strains were ESBL-positive, while ESBL-determining gene, blaTEM was detected in 23.8% of E. coli strains. The largest percentage of antibiotic resistant and MDR E. coli strains was detected nearby the STP, indicating that malfunctioning STP may contribute largely to river water contamination downstream, also having significant environmental and economic impact.

Antimicrobial resistance and ESBL genes in E. coli isolated in proximity to a sewage treatment plant.

This study was aimed to determine the prevalence of antibiotic-resistant Escherichia coli in the Szreniawa river with detailed aims of: (i) assessment of differences in the number of microbiological indicators of water quality in a diurnal cycle in a vicinity of the sewage treatment plant (STP); (ii) determination of prevalence of antimicrobial resistant E. coli isolated from three sites located at varying locations toward the STP; (iii) evaluation of the presence of extended-spectrum beta lactamase (ESBL)-determining genes in waterborne E. coli isolated from three sites of Szreniawa and (iv) genetic similarity assessment among the E. coli populations. Bacteriological contamination (coliforms, E. coli, E. faecalis) was assessed using membrane filtration. Fifty E. coli strains, the species of which was confirmed by MALDI-TOF analysis, were subjected to antimicrobial resistance tests using standard disk-diffusion method. Double disk synergy test was used to assess the ESBL production and PCR tests were conducted to detect the ESBL-conferring genes and evaluate the genetic diversity. A significant variation in the number of bacteriological indicators was observed both within and between the sampling sites, suggesting the effect of effluent from the STP, point discharge of household sewage and agricultural runoff on the water contamination. The resistance to amoxicillin/clavulanate (90%) and ampicillin (36%) was most prevalent. Multidrug resistance was observed in 40% of strains but no ESBL-producing strains were observed phenotypically. However, the presence of three ESBL-determining genes (TEM, OXA and CTX-M) was detected in 24, 10 and 8% of strains, respectively. A number of factors caused considerable pollution of the river and numerous multidrug resistant E. coli strains were isolated.

Assessment of microbiological aerosol concentration in selected healthcare facilities in southern Poland.

OBJECTIVES: This study was aimed to assess the concentration of microbial aerosol and species composition of airborne staphylococci in 10 healthcare facilities in southern Poland including primary healthcare units and hospital wards; and to assess whether the selected components of microbial aerosol pose a threat of severe infections to either patients or the personnel. METHODS: The study was conducted at monthly intervals over a period of one year. Air samples were collected by MAS-100 sampler. The number of mesophilic bacteria, mould fungi, actinomycetes and staphylococci was determined on general and selective media. The species identification of staphylococci was conducted using API tests for strains that were pre-selected based on macroscopic and microscopic observations. RESULTS: A total number of 1,584 samples were collected during the sampling period. The numbers of airborne microorganisms varied between the examined premises and between the seasons of the year. The observed differences were statistically significant with one exception for actinomycetes and their differences between the examined premises. The concentrations of mesophilic bacteria varied from 5 to 297 CFU/m3 of air, for Staphylococcus the values ranged from 1 to 96 CFU/m3, for fungi - from 1 to 100 CFU/m3, and the number of actinomycetes ranged from 7 to 321 CFU/m3. Ten species of coagulase-negative staphylococci (CoNS) were identified among 55 isolates with S. saprophyticus and S. warneri being the most frequently detected (n = 14 and 13, respectively). S. haemolyticus, which is one of the most common causal agents of nosocomial infections was observed in four facilities (n = 5). CONCLUSIONS: The microbial concentrations varied both between the seasons of the year and between the examined facilities. The highest bioaerosol concentrations were observed in most crowded premises. The identified species of staphylococci, although not typically associated with human infections, are common causal agents of nosocomial infections and infections in immunocompromised people.

Phenotypic and molecular assessment of antimicrobial resistance profile of airborne Staphylococcus spp. isolated from flats in Kraków.

Bacteria of the genus Staphylococcus were isolated from air sampled from living spaces in Kraków (Poland). In total, 55 strains belonging to the genus Staphylococcus were isolated from 45 sites, and 13 species of coagulase-negative staphylococci were identified. The species composition of studied airborne microbiota contains Staphylococcus species that are rarely infectious to humans. Most commonly isolated species comprised S. hominis and S. warneri. The disk-diffusion tests showed that the collected isolates were most frequently resistant to erythromycin. The PCR technique was employed to search for genes conferring the resistance in staphylococci to antibiotics from the group of macrolides, lincosamides and streptogramins. The analyzed Staphylococcus isolates possessed simultaneously 4 different resistance genes. The molecular analysis with the use of specific primers allowed to determine the most prevalent gene which is mphC, responsible for the resistance to macrolides and for the enzymatic inactivation of the drug by phosphotransferase. The second most often detected gene was msrA1, which confers the resistance of staphylococci to macrolides and is responsible for active pumping of antimicrobial particles out of bacterial cells.

Survival and Antibiotic Resistance of Bacteria in Artificial Snow Produced from Contaminated Water.

This study shows microbiological contamination of water in two main Podhale rivers, whose resources are used for the production of artificial snow, and the resulting snow contamination. Thirty-one E. coli strains were isolated from snow at two ski stations in the studied region, their antimicrobial resistance was determined, and the presence of extended spectrum ß-lactamase (ESBL) genes was searched for. The results indicate that the waters of both rivers are severely contaminated, resulting in the contamination of artificial snow with, among others, thermotolerant E. coli. E. coli isolated from snow were most frequently resistant to ampicillin (74.19%) and amoxicillin/clavulanate (51.61% isolates). Aminoglycosides and third generation cephalosporins were most efficient among the tested antimicrobials. Some bacterial strains were multidrug resistant and three strains exhibited the ESBL mechanism. Molecular analyses showed the presence of ESBL genes in the same three strains. Genetic variation among E. coli indicates that only some genotypes are able to survive the artificial snow production process.

Disc-diffusion and PCR detection of methicillin resistance in environmental airborne strains of Staphylococcus spp..

The aim of this study was to assess the species composition of airborne Staphylococcus spp. in public premises, to determine the methicillin resistance of the isolates and the prevalence of mecA gene, determining resistance to ß-lactams. In total 65 Staphylococcus strains were isolated from 54 sites. Four strains exhibited phenotypic methicillin resistance, while the presence of mecA gene was found in 11 strains. The results of both assays were compared, showing that the phenotypic tests revealed methicillin resistance only in 36% of the examined samples. This study revealed high species diversity among airborne Staphylococcus spp. population, which consists of multidrug resistant strains.

Assessment of microbiological and chemical properties in a municipal landfill area.

This study aimed at determining the environmental hazards for soils posed by a large municipal landfilll. The concentrations of heavy metals and Policyclic Aromatic Hydrocarbons, as well as microbial composition (i.e., mesophilic bacteria, actinomycetes, molds, Salmonella, Staphylococcus, Clostridium perfringens) in four soils within and in the vicinity of the landfill were evaluated and compared to waste samples. Both chemical and microbiological analyses revealed only limited contamination of surrounding areas. Although the increased alkalinity of soils was detected, the concentrations of heavy metals and Polycyclic Aromatic Hydrocarbons (PAHs) did not exceed the admissible values. All examined microbial groups were abundant in soil and waste. The highest microbial cell numbers were observed in warm summer and spring months. Although the site south of the landfill shows no trace of microbial contamination, pathogenic bacteria were found north of the landfill. This may suggest that there are other, more effective, transmission routes of bacteria than groundwater flow.

The molecular marker-based comparison of Azotobacter spp. populations isolated from industrial soils of Cracow-Nowa Huta steelworks (southern Poland) and the adjacent agricultural soils.

The occurrence of Azotobacter spp., which has beneficial effects on plant development, is related to various soil properties, such as pH and fertility. This study evaluated the prevalence of Azotobacter spp. in industrial (H) and agricultural soils (P) in Nowa Huta, Cracow and determined the phenotypic and genetic diversity of these bacteria. The examined bacteria were present in 40% of H and in 50% of P soils. Taxonomic identification of the bacterial isolates indicated the presence of three species--A. salinestris, A. chroococcum and A. vinelandii. The genetic diversity, determined using two fingerprinting methods--Random Analysis of Polymorphic DNA (RAPD) and Rep-PCR (BOX) revealed high level of population diversity. In AMOVA analysis most of diversity was attributed to within-population variation (76-85%), and only 3.78-6.18% was associated with among-group H and P variation. Global test of differences revealed distinct population structure within bacterial strains isolated from H and P areas only for BOX markers (Fst = 0.05732, P = 0.00275). Phenetic analyses: UPGMA and DCA better discriminated H and P groups based on RAPD data. Both BOX and RAPD methods provided an insight into the genetic complexity of Azotobacter spp. variation in soils of different land-use types.

The evaluation of heavy metal content in water and sediments of small reservoirs in light of various environmental quality regulations.

This article reports the heavy metal concentrations assessed both in bottom sediments and water flowing into two small reservoirs at Krempna and Zeslawice, southern Poland. The experiments were carried out in spring, summer, autumn and winter 2010-2011. The resulting concentrations of lead, cadmium, nickel, chromium and copper were compared with the Environmental Quality Standards, guidelines of the Regulation of the Minister of Environmental Protection, regulations of the Polish Geological Institute and State Institute of Environment Protection, Institute of Soil Science and Plant Cultivation, and with the regulations adopted in Germany, Denmark, the United States and Canada. Regarding the total heavy metal concentrations, water flowing into the examined reservoirs was not polluted. The highest concentrations of heavy metals were found in water flowing into the reservoirs in spring and summer. The heavy metal concentrations in sediments of the reservoirs at Krempna and Zeslawice were much higher than the concentrations of the elements discussed present in waters flowing into these reservoirs.

Second life of water and wastewater in the context of circular economy - Do the membrane bioreactor technology and storage reservoirs make the recycled water safe for further use?

In recent years water demand drastically increased which is particularly evident in tourism-burdened mountain regions. In these areas, climate neutral circular economy strategies to minimize human impact on the environment can be successfully applied. Among these strategies, treated wastewater reuse and retaining water in storage reservoirs deserve particular attention. This study aimed to determine if recycled water produced with two circular economy systems, namely membrane bioreactor treatment plant (MBR) with UV-light effluent disinfection and a storage reservoir, is safe enough for further use in green areas irrigation in summer and artificial snow production in winter. The assessment was based on the presence and concentration of antimicrobial agents, antibiotic resistant bacteria, antibiotic resistance genes, bacterial community composition and diversity. The treated water and wastewater was compared with natural water in their vicinity. Both systems fulfill the criteria set by the European Union in terms of reclaimed water suitable for reuse. Although the MBR/UV light wastewater treatment substantially reduced the numbers of E. coli and E. faecalis (from e.g. 32,000 CFU/100 ml to 20 CFU/100 ml and 15,000 CFU/100 ml to nearly 0 CFU/ml), bacteria resistant to ampicillin, aztreonam, cefepime, ceftazidime, ertapenem and tigecycline, as well as ESBL-positive and multidrug resistant E. coli were highly prevalent in MBR-treated wastewater (88.9 %, 55.6 %, 33.3 %, 22.2 % and 11.1 % and 44.4 and 55.6 %, respectively). Applying additional tertiary treatment technology is recommended. Retaining water in storage reservoirs nearly eliminated bacterial contaminants (e.g. E. coli dropped from 350 CFU/100 ml to 10 CFU/100 ml), antibiotic resistant bacteria, resistance genes (none detected in the storage reservoir) and antibiotics (only enrofloxacin detected once in the concentration of 3.20 ng/l). Findings of this study point to the limitations of solely culture-based assessment of reclaimed water and wastewater while they may prove useful in risk management and prevention in wastewater reuse.

The use of stem cells in the treatment of mastitis in dairy cows.

Mastitis is a multifactorial inflammatory disease. The increase in antibiotic resistance of bacteria that cause mastitis means that cattle breeders would prefer to reduce the use of antibiotics. Recently, therapies using mesenchymal stem cells (MSCs) from various sources have gained significant interest in the development of regenerative medicine in humans and animals, due to their extraordinary range of properties and functions. The aim of this study was to analyze the effectiveness of an allogeneic stem cells derived from bone marrow (BMSC) and adipose tissue (ADSC) in treating mastitis in dairy cattle. The research material consisted of milk and blood samples collected from 39 Polish Holstein-Friesian cows, 36 of which were classified as having mastitis, based on cytological evaluation of their milk. The experimental group was divided into subgroups according to the method of MSC administration: intravenous, intramammary, and intravenous + intramammary, and according to the allogeneic stem cells administered: BMSC and ADSC. The research material was collected at several time intervals: before the administration of stem cells, after 24 and 72 h, and after 7 days. Blood samples were collected to assess hematological parameters and the level of pro-inflammatory cytokines, while the milk samples were used for microbiological assessment and to determine the somatic cells count (SCC). The administration of allogeneic MSCs resulted in a reduction in the total number of bacterial cells, Staphylococcus aureus, bacteria from the Enterobacteriaceae group, and a systematic decrease in SCC in milk. The therapeutic effect was achieved via intravenous + intramammary or intramammary administration.

Waste Polyurethane Foams as Biomass Carriers in the Treatment Process of Domestic Sewage with Increased Ammonium Nitrogen Content.

In order to understand the mechanisms of microbial growth on waste polyurethane sponge materials, their effectiveness as biomass carriers in domestic sewage with increased ammonium nitrogen content treatment was assessed. Comparative experiments were carried out in microreactors under steady conditions of batch culture, which allowed for an assessment of different carriers, in the form of flexible foams, rigid foams, and flexible foams placed in full casings. In the studies conducted in continuous cultures, biomass carriers selected in batch culture were used as fillings in the column model. The structure of the microbial community inhabiting the spongy material was determined and the pollutant-removing process from real domestic sewage was assessed. Analyzes using the Illumina sequencing technique allowed for demonstrating that Nitrosomonas and Nitrospira were the predominant nitrifiers in the biomass carrier in the form of waste polyurethane foams (PUF). It was found that anammox bacteria, the presence of which-as unidentified Planctomycetes-was confirmed in the polyurethane sponge material, were also responsible for the high removal of N-NH4+. Burkholderia and Sphingopyxis phyla were identified as the dominant denitrifying bacteria involved in the treatment of domestic sewage with increased content of ammonium nitrogen. The biomass carrier in the form of waste PUF placed additionally in full casings proved to be more beneficial for the proliferation of bacteria involved in nitrification and denitrification processes. On the other hand, waste foams without casings proved to be more suitable for the growth of microorganisms known to perform partial denitrification and may accumulate nitrites (Staphylococcus, Dokdonella). Additionally, the presence of Devosia and Pseudonocardia, which participated in the phosphorus removal process, was found in the waste PUR foams.

Synthesis and Investigation of Physicochemical and Biological Properties of Films Containing Encapsulated Propolis in Hyaluronic Matrix.

The dynamic development of nanotechnology has enabled the development of innovative and novel techniques for the production and use of nanomaterials. One of them is the use of nanocapsules based on biodegradable biopolymer composites. Closing compounds with antimicrobial activity inside the nanocapsule cause the gradual release of biologically active substances into the environment, and the effect on pathogens is regular, prolonged and targeted. Known and used in medicine for years, propolis, thanks to the synergistic effect of active ingredients, has antimicrobial, anti-inflammatory and antiseptic properties. Biodegradable and flexible biofilms were obtained, the morphology of the composite was determined using scanning electron microscopy (SEM) and particle size was measured by the dynamic light scattering (DLS) method. Antimicrobial properties of biofoils were examined on commensal skin bacteria and pathogenic Candida isolates based on the growth inhibition zones. The research confirmed the presence of spherical nanocapsules with sizes in the nano/micrometric scale. The properties of the composites were characterized by infrared (IR) and ultraviolet (UV) spectroscopy. It has been proven that hyaluronic acid is a suitable matrix for the preparation of nanocapsules, as no significant interactions between hyaluronan and the tested compounds have been demonstrated. Color analysis and thermal properties, as well as the thickness and mechanical properties of the obtained films, were determined. Antimicrobial properties of the obtained nanocomposites were strong in relation to all analyzed bacterial and yeast strains isolated from various regions of the human body. These results suggest high potential applicability of the tested biofilms as effective materials for dressings to be applied on infected wounds.

Anthropogenic pollution gradient along a mountain river affects bacterial community composition and genera with potential pathogenic species.

Mountain regions in Poland are among the most frequently visited tourist destinations, causing a significant anthropogenic pressure put on the local rivers. In this study, based on numbers of 9 microorganisms, content of 17 antibiotics and 17 physicochemical parameters, we determined a pollution gradient in six sites along Bialka, a typical mountain river in southern Poland. The E.coli/Staphylococcus ratio varied evidently between polluted and non-polluted sites, indicating that the possible utility of this parameter in assessing the anthropogenic impact on river ecosystems is worth further investigation. Then, using next generation sequencing, we assessed the changes in bacterial community structure and diversity as a response to the pollution gradient. Proteobacteria and Bacteroidetes were the most abundant phyla in the majority of samples. Actinobacteria were the most abundant in the most pristine (groundwater) sample, while Firmicutes and Verrucomicrobia were more prevalent in polluted sites. Bacterial diversity at various levels increased with water pollution. Eleven bacterial genera potentially containing pathogenic species were detected in the examined samples, among which Acinetobacter, Rhodococcus, and Mycobacterium were the most frequent. At the species level, Acinetobacter johnsonii was most prevalent potential pathogen, detected in all surface water samples, including the pristine ones. Two bacterial taxa-genus Flectobacillus and order Clostridiales showed very distinct variation in the relative abundance between the polluted and non-polluted sites, indicating their possible potential as biomarkers of anthropogenic impact on mountain river waters.

Antibacterial Properties of Biodegradable Silver Nanoparticle Foils Based on Various Strains of Pathogenic Bacteria Isolated from the Oral Cavity of Cats, Dogs and Horses.

Frequent occurrence of microbial resistance to biocides makes it necessary to find alternative antimicrobial substances for modern veterinary medicine. The aim of this study was to obtain biodegradable silver nanoparticle-containing (AgNPs) foils synthesized using non-toxic chemicals and evaluation of their activity against bacterial pathogens isolated from oral cavities of cats, dogs and horses. Silver nanoparticle foils were synthesized using sodium alginate, and glucose, maltose and xylose were used as reducing agents. The sizes of AgNPs differed depending on the reducing agent used (xylose < maltose < glucose). Foil without silver nanoparticles was used as control. Bacterial strains were isolated from cats, dogs and horses by swabbing their oral cavities. Staphylococcus aureus, methicillin-resistant Staphylococcus aureus (MRSA), Escherichia coli and extended-spectrum beta-lactamase (ESBL) producing E. coli were isolated on selective chromogenic microbiological media. The bactericidal effect of AgNPs foils obtained using non-toxic chemical compounds against E. coli, ESBL, S. aureus and MRSA isolated from oral cavities of selected animals was confirmed in this study. No statistically significant differences were observed between the foils obtained with different reducing agents. Therefore, all types of examined foils proved to be effective against the isolated bacteria.