The effect of biological methods for MSW treatment on the physicochemical, microbiological and phytotoxic properties of used biofilter bed media.

Biofilters are commonly used in municipal solid waste treatment (MSW) facilities to remove odors and pollutants from process gases. However, the effectiveness of biofilter bed media decreases over time, necessitating periodic replacement. The type of the treatment process may affect the lifespan of the bed and the way it should be utilized after replacement. This study aimed to analyze the physical, chemical, calorific, microbiological, and phytotoxic parameters of bed media in biofilters operated at an industrial scale in MSW treatment plants. The experiments included three full cycles of biofiltering gases from biodrying, composting, and aerobic biostabilization in two variations. Physicochemical properties (moisture, organic matter, carbon, nitrogen, sulfur, heavy metal contents), respiration activity (AT4), phytotoxicity, and microorganism abundance were determined for initial materials and samples from two biofilter layers collected after each cycle. Results revealed a substantial reduction in AT4 (by 63%-87% compared to initial material), significant moisture content increase in the bottom layers (by 61% or more, depending on the process), and a considerable decrease in microorganism abundance. Biofilter bed media from biodrying and composting exhibited low environmental risk (low heavy metal concentrations, negligible phytotoxicity, and microbiological stability). However, bed packings from aerobic biostabilization processes showed significant inhibition of indicator plants and incomplete sanitization (presence of pathogens like E. coli and Salmonella spp.). Therefore, these bed packings can be utilized for energy recovery, such as incineration after drying. This research provides significant insights into the effectiveness and safety of biofilter bed media in MSW treatment plants.

Impact of digestate addition on the biostabilization of undersized fraction from municipal solid waste.

Biostabilization is a commonly applied method in mechanical-biological treatment (MBT) plants to process municipal solid waste. In many ways, e.g. by applying additives to waste, MBT plant operators strive to enhance the effectiveness of biostabilization, which leads to reducing the time and energy outlays necessary for the process, as well as to minimizing the amount of final stabilized waste directed to landfills. This paper deals with the impact of digestate waste from agricultural biogas plants used as additive to the biostabilization process of undersized fraction from municipal solid waste (UFMSW) on the intensive phase of the process and properties of stabilized waste. The aim of this study was to assess whether, and if so to what extent, the application of digestate waste affects the process. Five different input compositions were tested (without digestate and with the addition of digestate at: 2.5; 5; 7.5 and 10 wt%). Waste treatment time was 2 weeks. Changes in moisture content, organic matter (OM), respiration activity (AT4), bulk density, air-filled porosity, heavy metal content, pH, carbon to nitrogen ratio, as well as composition of process gases emitted were evaluated. Additionally, microorganisms (including pathogens) inhabiting the processed waste in the aspect of waste sanitation were analyzed. It was found that the addition of digestate at 2.5, 5 and 7.5 wt% extended the duration of the thermophilic phase and decreased the CO2 content in process gases. The addition of digestate at 2.5 wt% and 5 wt%, decreased also OM by approx. 25% of the initial value and AT4 by approx. 30%. It was also proved that the addition of digestate favors the limited sanitation of UFMSW. As a result of the research, it was found that the addition of digestate at 2.5 wt% and 5 wt% is sufficient to accelerate the aerobic biological degradation of UFMSW.

Rheological and Microbiological Characteristics of Hops and Hot Trub Particles Formed during Beer Production.

During the production of beer, and especially beer wort, the main wastes are spent grain and hot trub, i.e., the so-called "hot break." Combined with yeast after fermentation, they represent the most valuable wastes. Hot trub is also one of the most valuable by-products. Studies on the chemical composition of these sediments and their rheological properties as waste products will contribute to their effective disposal and even further use as valuable pharmaceutical and cosmetic raw materials. So far, hot trub has been studied for morphology and particle distribution depending on the raw material composition and beer wort extract. However, there are no preliminary studies on the rheological properties of hot trub and hops. In particular, no attention has yet been paid to the dependence of these properties on the hop variety or different protein sources used. The aim of this study was to examine the effect of different hopping methods on hot trub viscosity and beer wort physicochemical parameters. Additionally, the hop solutions were measured at different temperatures. A microbiological analysis of hop sediments was also performed to determine the post-process survival of selected microorganisms in these wastes. For manufacturers of pumps used in the brewing industry, the most convenient material is that of the lowest viscosity. Low viscosity hot trub can be removed at lower velocities, which reduces costs and simplifies washing and transport. The sediments also had similar equilibrium viscosity values at high shear rates.

Impact of Calcium Oxide on Hygienization and Self-Heating Prevention of Biologically Contaminated Polymer Materials.

During the storage of spent polymer materials derived from municipal solid waste, which contain biodegradable impurities, an intense growth of microorganisms takes place. The aerobic metabolism of microorganisms may cause these materials to combust spontaneously and to become a real epidemiological risk for humans. The aim of the research is to determine the optimal addition of calcium oxide (CaO), which effectively reduces the number of selected microorganism groups populating the analyzed materials, in which spent polymers represent a significant fraction: refuse-derived fuel (RDF) and an undersize fraction of municipal solid waste (UFMSW). The main novelty of the experiments is to assess the benefits of using the commonly available and cheap filler (CaO), to hygienize the material and to reduce the fire hazard arising from its storage. During the mixing of spent polymer materials with pulverized CaO (mass shares: 1, 2, and 5% CaO), temperature changes were monitored using thermography. Moisture content (MC), pH, respiration activity (AT4) and bacterial count were determined before and after the experiment. During the addition of CaO (especially when the content was at 5%) to the UFMSW, higher maximum temperatures were obtained than in the case of RDF analyses, which may be the result of a high percentage of the biodegradable fraction and higher MC of the UFMSW. In all cases the waste temperature did not increase again after 3 min. CaO used in the experiment effectively limited the number of microorganisms. The addition of 5% of CaO has showed the strongest antimicrobial properties, and it can be recommended for hygienization of the analyzed materials and for the reduction of the risk of self-heating during their storage in windrows.

Effect of biochar addition on the OFMSW composting process under real conditions.

The article evaluates the effect of small selected doses of biochar addition (0%, 1.5%, 3% and 5%, wet weight) on the composting process of the organic fraction of municipal solid waste (OFMSW) with a low initial C/N ratio under real conditions. The low C/N composting mixtures with addition of biochar at low rates can have a positive effect on the compost quality and on the reduction of N losses in compost. The novelty of this work consists in studying the impact of small biochar doses on the composting process at full-scale. The research was conducted under real conditions in the Brno Central Composting Plant (Czech Republic) receiving food waste, grass, straw, sawdust, mineral waste, paper, wood and sewage sludge for processing. The experimental processing time was 12 weeks. We evaluated changes in carbon (C), nitrogen (N), moisture content (MC), organic matter (OM), respiration activity (AT4), as well as changes in the microbiocenotic composition of microorganisms colonizing the processed waste. OFMSW with the addition of biochar and compost were assessed for the content of heavy metals (HM). It was found out that biochar reduced the compost toxicity. The resulting compost with the addition of biochar exhibited higher moisture content and lower waste density. Biochar had an impact on N retention during composting but it did not change the course or accelerate the composting process. The highest OM loss (62.6%) was observed in the OFMSW with no biochar addition. The abundance of potentially pathogenic microorganisms clearly decreased during the OFMSW composting process with the addition of biochar.

Correction to: Antimicrobial resistance and the presence of extended-spectrum beta-lactamase genes in Escherichia coli isolated from the environment of horse riding centers.

The original publication of this paper contains a mistake.

Eco-friendly approach to the synthesis of silver nanoparticles and their antibacterial activity against Staphylococcus spp. and Escherichia coli.

The aim of this study was to assess bactericidal properties of nanosilver obtained with ascorbic acid (vitamin C) as a reducing substance, against environmental strains of Gram-positive (Staphylococcus) and Gram-negative bacteria (Escherichia coli). Silver nanoparticles were obtained by a simple and fast method of chemical reduction in mild synthesis condition using substrates not classified as dangerous and commonly found in plants. Bacterial susceptibility to nanosilver was determined using the disk-diffusion method. Gram-positive bacteria were less susceptible to bactericidal action of nanosilver and minimum inhibitory concentration was higher for these bacteria. However, susceptibility of individual isolates of different species to nanosilver was very diverse. The range of growth inhibition zones indicates that the resistance to different concentrations of nanosilver was a strain-characteristic, not species-related feature. The study of effective nanomaterials synthesis and applicability appears to be an extremely important element in the development of nanotechnology, especially in terms of the risks assessment. Obtained results have a real chance to be an important step toward creating a new generation of disinfectants without toxic effects on the environment and higher organisms.

Antimicrobial resistance and the presence of extended-spectrum beta-lactamase genes in Escherichia coli isolated from the environment of horse riding centers.

The aim of the study was to determine the antimicrobial resistance profile and the occurrence of extended-spectrum beta-lactamase genes and to analyze the genetic diversity of Escherichia coli strains isolated from the environment of horse riding centers. The study was conducted using E. coli strains isolated from the air, manure, and horse nostril swabs in three horse riding centers differing in the system of horse keeping-stable (OJK Pegaz and KJK Szary) and free-range (SKH Nielepice). Resistance to antibiotics was determined using the disk-diffusion method, and the PCR technique was employed to detect the extended-spectrum ß-lactamase (ESBL) genes, while the genetic diversity of strains was assessed by rep-PCR. A total of 200 strains were collected during the 2-year study, with the majority isolated from KJK Szary, while the smallest number was obtained from SKH Nielepice. The strains were mostly resistant to ampicillin, aztreonam, and ticarcillin. The tested strains were most frequently resistant to one or two antibiotics, with a maximum of ten antimicrobials at the same time. Two multidrug-resistant (MDR) strains were detected in OJK Pegaz while in KJK Szary there were two MDR and one extensively drug-resistant (XDR) strain. The ESBL mechanism was most frequently observed in OJK Pegaz (20.31% of strains) followed by KJK Szary (15.53% of strains) and SKH Nielepice (15.15% of strains). Among the ESBL-determining genes, only blaTEM and blaCTXM-9 were detected-blaTEM was mostly found in KJK Szary (53.40% of strains), while the second detected gene-blaCTXM-9-was most frequent in SKH Nielepice (6.06% of strains). The rep-PCR genotyping showed high variation among the analyzed strains, whereas its degree differed between the studied facilities, indicating that the type of horse keeping (stable vs. free-range) affects the genetic diversity of the E. coli strains. Having regard to the fact that the tested strains of E. coli were derived from non-hospitalized horses that were not treated pharmacologically, we can assume that the observed antimicrobial resistance may be of both-natural origin, i.e., not the result of the selection pressure, and acquired, the source of which could be people present in the horse riding facilities, the remaining horses which were not included in the study, and air, as well as water, fodder, and litter of the animals. Therefore, it can be concluded that the studied horses are the source of resistant E. coli and it is reasonable to continue monitoring the changes in antimicrobial resistance in those bacteria.

Microbiological quality of air in free-range and box-stall stable horse keeping systems.

The aim of this study was to assess the microbiological quality of air in three horse riding centers differing in the horse keeping systems. The air samples were collected in one facility with free-range horse keeping system and two with box stalls of different sizes. The samples were collected over a period of 3 years (2015-2017), four times per year (spring, summer, autumn, winter) to assess the effect of seasonal changes. The prevalence of aerobic mesophilic bacteria, mold fungi, actinomycetes, Staphylococcus spp., and Escherichia coli was determined by the air collision method on Petri dishes with appropriate microbiological media. At the same time, air temperature, relative humidity, and particulate matter concentration (PM10, PM2.5) were measured. It was found that the horse keeping system affects the occurrence of the examined airborne microorganisms. Over the 3-year period of study, higher temperature and humidity, as well as particulate matter concentration-which notoriously exceeded limit values-were observed in the facilities with the box-stall system. The air sampled from the largest horse riding center, with the largest number of horses and the box-stall system of horse keeping, was also characterized by the heaviest microbiological contamination. Among others, bacteria from the following genera: Staphylococcus spp., Streptococcus spp., Bacillus spp., and E. coli and fungi from the genera Aspergillus, Fusarium, Mucor, Rhizopus, Penicillium, Trichothecium, Cladosporium, and Alternaria were identified in the analyzed samples.

Silver nanoparticles toxicity against airborne strains of Staphylococcus spp.

The aim of this study was to explore the toxicity of silver nanoparticles (AgNPs) synthesized by chemical reduction method assessment with regard to airborne strains of Staphylococcus spp. The first step of the experiment was the preparation of silver nanoparticle suspension. The suspension was obtained by a fast and simple chemical method involving the reduction of silver ions through a reducing factor in the presence of the suitable stabilizer required to prevent the aggregation. In the second stage, varied instrumental techniques were used for the analysis and characterization of the obtained nanostructures. Third, the bacteria of the Staphylococcus genus were isolated from the air under stable conditions with 47 sports and recreational horses, relatively. Next, isolated strains were identified using biochemical and spectrophotometric methods. The final step was the evaluation of the Staphylococcus genus sensitivity to nanosilver using the disk diffusion test. It has been proven that prepared silver nanoparticles exhibit strong antibacterial properties. The minimum inhibitory concentration for tested isolates was 30 µg/mL. It has been found that the sensitivity of Staphylococcus spp. isolated from six identified species differs considerably. The size distribution of bacterial growth inhibition zones indicates that resistance to various nanosilver concentrations is an individual strain feature, and has no connection with belonging to a specific species.

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.

Phenotypic and Molecular Assessment of Drug Resistance Profile and Genetic Diversity of Waterborne Escherichia coli.

Bacterial Escherichia coli isolates were derived from waters of the Nowohucki Reservoir (Cracow, Poland) in summer and winter seasons. In total, 94 strains, identified as E. coli, were isolated from five sampling sites in the area of the reservoir. Based on the disk-diffusion tests, it was found that the tested isolates were predominantly resistant to ticarcillin and ampicillin. Numerous multi-drug resistant strains were detected, which however did not exhibit the ESBL phenotype. However, PCR approach allowed to detect ESBL-mechanism genes (CTX-M3, OXA, SHV, and TEM) in as much as 38 % of E. coli isolates. These results were coupled with significant molecular diversity of the E. coli strains revealed in BOXA1R-based rep-PCR technique.

Phenotypic and Molecular Antibiotic Resistance Determination of Airborne Coagulase Negative Staphylococcus spp. Strains from Healthcare Facilities in Southern Poland.

This study assessed the antimicrobial resistance of airborne Staphylococcus spp. strains isolated from healthcare facilities in southern Poland. A total of 55 isolates, belonging to 10 coagulase-negative staphylococci (CoNS) species, isolated from 10 healthcare facilities (including hospitals and outpatient units) were included in the analysis. The most frequently identified species were Staphylococcus saprophyticus and Staphylococcus warneri, which belong to normal human skin flora, but can also be the cause of common and even severe nosocomial infections. Disk diffusion tests showed that the bacterial strains were most frequently resistant to erythromycin and tetracycline and only 18% of strains were susceptible to all tested antimicrobials. Polymerase chain reaction amplification of specific gene regions was used to determine the presence of the Macrolide-Lincosamide-Streptogramin resistance mechanisms in CoNS. The molecular analysis, conducted using specific primer pairs, identified the msrA1 gene, encoding active efflux pumps in bacterial cells, as the most frequent resistance gene. As many as seven antibiotic resistance genes were found in one isolate, whereas the most common number of resistance genes per isolate was five (n = 17). It may be concluded that drug resistance was widely spread among the tested strains, but the resulting antimicrobial resistance profile indicates that in the case of infection, the use of antibiotics from the basic antibiogram group will be effective in therapy. However, before administering treatment, determination of the specific antimicrobial resistance should be conducted, particularly in the case of hospitalized patients.

The prevalence of selected genes involved in the biosynthesis of trichothecenes assessed with the specific PCR tests in Fusarium spp. isolated from cereals in southern Poland.

The analysis was conducted using 50 isolates of fungi of the genus Fusarium belonging to the species classified as major trichothecene mycotoxin producers: F. graminearum, F. culmorum, F. sporotrichioides, and F. poae. The tested fungi were isolated from ears of cereal crops in southern Poland during the two growing seasons (2011 and 2012). The aim of this study was to evaluate the prevalence of genes involved in the biosynthesis of trichothecene mycotoxins using the specific PCR tests. Molecular analyses indicated that the genes responsible for the production of trichothecenes (Tri3, Tri5, Tri7, Tri13) were abundant in the examined genetic material. The tested fungal isolates were characterized by a large diversity in terms of the number and composition of the possessed Tri genes. On the other hand, 14 of 50 isolates were found not to carry any of Tri genes.

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.

In vitro effects of various xenobiotics on Fusarium spp. strains isolated from cereals.

This study aimed to determine the susceptibility of Fusarium spp. strains isolated from cereals to selected heavy metals, fungicides and silver nanoparticles. The experiments were conducted using 50 Fusarium strains belonging to five species: F. graminearum, F. culmorum, F. oxysporum, F. sporotrichioides and F. avenaceum. The strains were found to be highly resistant to Pb(2+) and Zn(2+). Medium resistance to Cu(2+) and Mn(2+) and low resistance to Cd(2+) and Fe(3+) was also observed. Among the tested fungicides, formulations containing azoxystrobin, prochloraz and tebuconazole proved to be the most effective in inhibiting the growth of fungi, as they affected fungal growth in each of the applied doses. Susceptibility of Fusarium spp. to nanosilver, demonstrated in this study, shows the legitimacy of using nanostructures as fungicidal agents. The results confirm high diversity of the analyzed fungal species in terms of susceptibility to the tested substances, and encourage to continue research on the resistance of Fusarium spp. to various fungicidal agents.

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.

[Evaluation of ITS-PCR and PCR MP techniques for Streptococcus agalactiae genetic differentiation]. / Ocena przydatnosci metod: ITS-PCR i PCR MP w genotypowaniu Streptococcus agalactiae.

INTRODUCTION: The aim of this study was to evaluate the usefulness of ITS-PCR and PCR MP methods for genotyping of S. agalactiae strains isolated from women in reproductive age. METHODS: In the course of the study 250 strains of S. agalactiae were isolated and their serotype was identified. The ITS-PCR and PCR MP methods allowed to differentiate 20 strains and then the correlation between the serotypes of the tested isolates and their geno- type was evaluated. RESULTS: Among 250 strains of S. agalactiae the following serotypes were identified as the most common: III (54%), Ia (17%) and V (12%). Also other serotypes have been found: IV (8%), Ib (5%) and II (4%). PCR MP has a higher discriminative power than the ITS-PCR and it allowed for the efficient differentiation of strains. There is no direct relationship between genotypes and serotypes ofS. agalactiae. CONCLUSIONS: PCR MP is useful in the differentiation of S. agalactiae. Exactly as in the case of RAPD and PFGE methods, PCR MP can be used for thediagnosis and analysis of GBS isolates colonization. This method is also used for successful genotyping of other microbial species, often closely related ones, as well as in epidemiological studies.

The effect of heavy metal concentration and soil pH on the abundance of selected microbial groups within ArcelorMittal Poland steelworks in Cracow.

The present study aimed to identify the effect of heavy metal concentration and soil pH on the abundance of the selected soil microorganisms within ArcelorMittal Poland steelworks, Cracow. The analysis included 20 soil samples, where the concentration of Fe, Zn, Cd, Pb, Ni, Cu, Mn, Cr and soil pH were evaluated together with the number of mesophilic bacteria, fungi, Actinomycetes and Azotobacter spp. In the majority of samples soil pH was alkaline. The limits of heavy metals exceeded in eight samples and in one sample, the concentration of Zn exceeded 31-fold. Chromium was the element which most significantly limited the number of bacteria and Actinomycetes.