53 soil samples collected in the Bellsund Region in Western Spitsbergen were examined. An acid-resistant strain difficult to identify was isolated and recognized as Mycobacterium friburgensis. 54.7% of isolated strains were acid-resistant and growing at 25˚C only. They were microorganisms at borderline of Mycobacteria and Actinomycetes. Other microorganisms isolated in the studied soil samples were bacilli (55.7%) and coccaceae (15.4%).
Total count (TC) of bacteria in drifting annual pack-ice in austral spring fluctuated between 2.8-106 and 2,09-109 dm3. TC of bacteria was lowest in the upper layer of a large pack-ice fragment, emersed above water surface and almost completely free of diatoms; it was comparable to TC of bacteria in surrounding sea water, which was very low at this time (1,92- 106 — 5.8-106 dm -3). TC of bacteria increased in the deeper layers of pack-ice, attaining a maximum in the middle layer characterized by a high count of diatoms. TC of bacteria was highest in small pack-ice pieces 10—20 kg in being and densely overgrown with diatoms. Bacterial population in pack-ice was dominated by rods (62%), and it contained filamentous bacteria (2.4%) and prosthecate forms (4,8%), rarely present in deep sea. Mean volume of bacterial cell (0,206/μm3) was small, only slightly exceeding that of cells of free-living bacteria in sea water in summer.
Water samples were collected at 12 oceanographic stations from six standard depths ranging from 0 to 100 and 150 m. The number of bacteria and concentration of organic components were expressed in adequate units per 1 litre of sea water and in the form of the integrated values for the whole water column under I m2 of sea of organic components were expressed in adequate units per 1 litre of sea water and in the form of the integrated values for the whole water column under 1 m2 of sea surface. Total numbers of bacteria (TC) ranged from 0.16 to 7.31 x 107/1 and 1.74 — 5.67 x 10, 2/m2 saprophytic bacteria (CFU) 0.10 — 46.85 x 103/1 and 0.62 — 27.7x 108/m2. contents of particulate organic carbon (РОС) 0.02 — 0.25 mg/1 and 3.5 — 20.0 g/m2 dissolved organic carbon (DOC) 0.07 — 3.02 mg/1 and 53.5 — 207.9 g/m2, dissolved free amino acids (DFAA) 0 — 1.8965 μmol/1 and 2.7 -151.5 mmol/m2, dissolved combined amino acids (DCAA) 0 2.9366 μmо1/1 and 16.5— 163.5 mmol/m2, particulate combined amino acids (PCAA) 0 — 3.0215 μmо1/1 and 3.7 — 249.0 mmol/m2. Total numbers of bacteria and РОС, DOC and DCAA concentrations, widely differentiated in the investigated area, were on the average much lower than the values obtaine in previous years. The saprophytic bacteria content and DFAA and PCAA concentrations were at a similar level to that in the past years. Higher TC and CFU values were observed in the areas with high concentrations of phytoioplankton to the NW of Anvers I. and around Clarence I.
The total numbers and biomass of bacterioplankton in two Arctic glacial fjords off west Spitsbergen were studied. Samples were collected from different water depth layers – from the surface to 80–90 m depth. Total bacterial number (TBN), biomass and morphological structure (shape of bacteria) were determined using the acridine orange direct count method. The highest values of TBN and biomass in the water column were found in Kongsfjorden in the stations adjacent to Kongsbreen Glacier, and the lowest values in the outer part of the Krossfjorden. The local maxima of bacterioplankton were observed in water layers around pycnocline. The morphological structure was similar in all samples – the bacteria were dominated by rods (over 65%), followed by cocci (16–20%) and vibrios (11–15%).
The paper presents the first physicochemical and microbiological studies conducted in the northern area of Svalbard (Spitsbergen). Ten sediment samples were collected from the bottom of the longest fjord in the region, Wijdef jorden. Bottom sediments from ten lakes located along the shores of Wijdefjorden and Woodfjorden were also sampled. Organic matter content (LOI), water content, temperature, pH, and salinity of the sediments were determined. The quantity of aerobic bacteria cultured on various growth media at 4 ° C, 14 ° C, and 37 ° C ranged from 10 2 to 10 6 cfu/g of wet sediment mass, depending on the type of sampling station (fjord or lake). The number of bacteria did not co rrelate with organic matter content. Out of the 37 bacterial strains isolated from Wijdefjorden, 48% and 70% revealed ureolytic and proteolytic activity, respectively. The proportion of freshwater strains with ureolytic and proteolytic activity was 32% and 55%, respectively. Antibiotic resistance testing indicated that bacterial strains from the bottom sediments of the lakes were resistant to 8 antibiotics (out of the 18 investigated). Possible sources of this resistance are discussed. Using 16S DNA analysis, bacterial isolates from the lakes were identified as Pseudomonas sp., whereas frequently occurring strains in bottom sediment of the fjord were Pseudoalteromonas sp.
The research was focused on the selection of the best conditions for the lactic acid production. As the organic source diluted waste whey was used. Two facultative anaerobic bacteria strains were examined: Lactobacillus rhamnosus and Lactococcus lactis. The neeed of anaerobic conditions as well as mineral supplementation of cultivationwere investigated. It turned out that the oxidationwas not the key parameter, but cultivationmediumneeded a supplementation for higher process efficiency. Finally, Lactobacillus rhamnosus strain was selected, for which LA production was app. 45% higher than for Lc. lactis. On the other hand, Lactobacillus rhamnosus was active at higher lactose concentration, thus waste whey needed to be less diluted. Additionally, high values of product/substrate yield coefficient make the process very efficient.
Effects of fermented extruded rye flour supplements with Lactobacillus sakei KTU05-6 or Pediococcus pentosaceus BaltBio02 on milk production and composition, as well as ruminal parameters, were determined in Lithuanian Black & White dairy cows. Also, determination of antimicrobial activities of tested lactic acid bacteria (LAB) against a variety of pathogenic and opportunistic bacterial strains previously isolated from diseased cattle was performed. The highest antimicrobial activity was demonstrated in L. sakei against S. aureus, and in P. pentosaceus against P. aeruginosa and S. aureus. The count of LAB in the supplements after 72 h of fermentation of extruded rye flour with L. sakei and P. pentosaceus was 9.6±0.4 log10 CFU/g and 9.5±0.3 log10 CFU/g, respectively. All cows (n=60) were fed the same basal diet. The treatment differences were achieved by individually incorporating (65 d.) one of the supplements: L. sakei KTU05-6 (group B; n=20) or P. pentosaceus BaltBio02 (group C; n=20). The control group A (n=20) was on the basal diet only. A supplement fermented with L. sakei does not have a significant influence on dairy cattle milk production and rumen fluid parameters. The type of LAB used has a significant influence (p<0.0001) on microbiological parameters of the rumen (TCM, TCL, TCE). The milk yield was increased (p≤0.05) using P. pentosaceus BaltBio02 supplement, and further research is needed to identify w hat is the main mechanism of the positive action.
Water is a strategic material. Recycling is an important component of balancing its use. Deep-bed filtration is an inexpensive purification method and seems to be very effective in spreading water recovery. Good filter designs, such as the fibrous filter, have high separation efficiency, low resistance for the up-flowing fluid and high retention capacity. However, one of the substantial problems of this process is the biofouling of the filter. Biofouling causes clogging and greatly reduces the life of the filter. Therefore, the melt-blown technique was used for the formation of novel antibacterial fibrous filters. Such filters are made of polypropylene composites with zinc oxide and silver nanoparticles on the fiber surface. These components act as inhibitors of bacterial growth in the filter and were tested in laboratory and full scale experiments. Antibacterial/bacteriostatic tests were performed on Petri dishes with E. coli and B. subtilis. Full scale experiments were performed on natural river water, which contained abiotic particles and mutualistic bacteria. The filter performance at industrial scale conditions was measured using a particle counter, a flow cytometer and a confocal microscope. The results of the experiments indicate a significant improvement of the composite filter performance compared to the regular fibrous filter. The differences were mostly due to a reduction in the biofouling effect.
Cold−adapted marine bacteria producing extracellular hydrolytic enzymes are important for their industrial application and play a key role in degradation of particulate or ganic matter in their natural environment. In this work, members of a previously−obtained protease−producing bacterial collection isolated from different marine sources from Potter Cove (King George Island, South Shetlands) were taxonomically identified and screened for their ability to produce other economically relevant enzymes. Eighty−eight proteolytic bacterial isolates were grouped into 25 phylotypes based on their Amplified Ribosomal DNA Restriction Analysis profiles. The sequencing of the 16S rRNA genes from representative isolates of the phylotypes showed that the predominant culturable protease−producing bacteria belonged to the class Gammaproteobacteria and were affiliated to the genera Pseudomonas , Shewanella , Colwellia , and Pseudoalteromonas , the latter being the predominant group (64% of isolates). In addition, members of the classes Actinobacteria, Bacilli and Flavobacteria were found. Among the 88 isolates screened we detected producers of amylases (21), pectinases (67), cellulases (53), CM−cellulases (68), xylanases (55) and agarases (57). More than 85% of the isolates showed at least one of the extracellular enzymatic activities tested, with some of them producing up to six extracellular enzymes. Our results confirmed that using selective conditions to isolate producers of one extracellular enzyme activity increases the probability of recovering bacteria that will also produce additional extracellular enzymes. This finding establishes a starting point for future programs oriented to the prospecting for biomolecules in Antarctica.
The aim of this research was to evaluate the microbiological indoor air contamination level in chosen facilities of the primary health-care for adults and children. The total numbers of mesophilic bacteria, staphylococci, coli-group bacteria and moulds in both surgery rooms and patients’ waiting rooms were determined. Air samples were collected with a MAS 100 impactor and the concentration of microorganisms was estimated by a culture method. The microbiological air contamination level was diverse: the number of mesophilic bacteria ranged from 320 to 560 CFU/m3, number of staphylococci - 10-305 CFU/m3, coli group bacteria - 0-15 CFU/m3 and moulds - 15-35 CFU/m3. The bacteriological contamination level of the air in examined community health centers was higher than described in the literature for hospitals and exceeded the acceptable values proposed for the surgery objects.
In general, Antarctic marine bacteria are small, with biovolumes ranging from 0.139 to 0.204 μm-3 cell-1, but their total biomass in seawater is considerable due to relatively high numbers that approximate to 1020 cells km-3. Bacterial biomass becomes more concentrated closer to land. Our multi-year Antarctic studies demonstrated an average total bacterial biomass of 504 tons in Admirality Bay (24 km3) or 21 tons per 1 km3, versus 6.4 tons per 1 km3 in the open ocean. Strikingly, bacterial biomass reached 330 tons per 1 km3 of seawater at the sea-ice edge, as sampled in Goulden Cove in Admiralty Bay. Bacterial biomass in Admirality Bay, which we believe can be enriched by halotolerant and thermotolerant fresh water bacteria from glacial streams, is equal to or even exceeds that of the standing stock of krill (100-630 tons per bay) or other major living components, including phytoplankton (657 tons), flagellates (591 tons), and ciliates (412 tons). However, the bacterial biomass is exceeded by several orders of magnitude by non-living organic matter, which constitutes the basic bacterial carbon source. Factors regulating high bacterial abundance in the vicinity of land are discussed.
We studied dynamic changes in anthropogenic bacterial communities at a summer−operated Czech research base (the Mendel Research Station) in the Antarctic during 2012 and 2013. We observed an increase in total numbers of detected bacteria between the beginning and the end of each stay in the Antarctic. In the first series of samples, bacteria of Bacillus sp. predominated. Surprisingly, high numbers of Gram−positive cocci and coli − forms were found (including opportunistic human pathogens), although the conditions for bacterial life were unfavourable (Antarctic winter). In the second series of samples, coliforms and Gram−positive cocci predominated. Dangerous human pathogens were also detected. Yersinia enterocolitica was identified as serotype O:9. Antibiotic susceptibility testing showed medium−to−high resistance rates to ampicillin, cefalotin, cefuroxime, amoxicillin−clavulanate and gentamicin in Enterobacteriaceae. 16S rRNA sequencing showed high rates of accordance between nucleotide sequences among the tested strains. Three conclusions were drawn: (1) Number of anthropogenic bacteria were able to survive the harsh conditions of the Antarctic winter (inside and outside the polar station). Under certain circumstances ( e.g. impaired immunity), the surviving bacteria might pose a health risk to the participants of future expeditions or to other visitors to the base. (2) The bacteria released into the outer environment might have impacts on local ecosystems. (3) New characteristics ( e.g. resistance to antibiotics) may be introduced into local bacterial communities.
In this study, selected heavy metals resistant heterotrophic bacteria isolated from soil samples at the Windmill Islands region, Wilkes Land (East Antarctica), were characterized. Phylogenetic analysis revealed affiliation of isolates to genera Bacillus , Lysinibacillus , Micrococcus and Stenotrophomonas . The strains were found to be psychrotolerant and halotolerant, able to tolerate up to 10% NaCl in the growth medium. The Minimum Inhibitory Concentration of the seven heavy metals Cr, Cu, Ni, Co, Cd, Zn, and Pb was deter − mined in solid media for each bacterial strain. Gram−positive Vi−2 strain and Gram−negative Vi−4 strain showed highest multiply heavy metals resistance, and Vi−3 and Vi−4 strains showed multi−antibiotic resistance to more than a half of the 13 used antibiotics. Plasmids were detected only in Gram−negative Vi−4 strain. The bacteria were able to produce different hydrolytic enzymes including industrially important proteases, xylanases, cellulases, and b −glucosidases. High heavy metals resistance of the Antarctic bacteria suggests their potential application for wastewater treatment in cold and temperate climates. Highly sensitive to Cd and Co ions Vi−1, Vi−5 and Vi−7 strains would be promising for developing biosensors to detect these most toxic heavy metals in environmental samples.
MDAP-2 is a new antibacterial peptide with a unique structure that was isolated from house- flies. However, its biological characteristics and antibacterial mechanisms against bacteria are still poorly understood. To study the biological characteristics, antibacterial activity, hemolytic activi- ty, cytotoxicity to mammalian cells, and the secondary structure of MDAP-2 were detected; the results showed that MDAP-2 displayed high antibacterial activity against all of the tested Gram-negative bacteria. MDAP-2 had lower hemolytic activity to rabbit red blood cells; only 3.4% hemolytic activity was observed at a concentration of 800μg/ml. MDAP-2 also had lower cytotoxicity to mammalian cells; IC50 values for HEK-293 cells, VERO cells, and IPEC-J2 cells were greater than 1000 μg/ml. The circular dichroism (CD) spectra showed that the peptide most- ly has α-helical properties and some β-fold structure in water and in membrane-like conditions. MDAP-2 is therefore a promising antibacterial agent against Gram-negative bacteria. To deter- mine the antibacterial mechanism(s) of action, fluorescent probes, flow cytometry, and transmis- sion electron microscopy (TEM) were used to study the effects of MDAP-2 on membrane perme- ability, polarization ability, and integrity of Gram-negative bacteria. The results indicated that the peptide caused membrane depolarization, increased membrane permeability, and destroyed membrane integrity. In conclusion, MDAP-2 is a broad-spectrum, lower hemolytic activity, and lower cytotoxicity antibacterial peptide, which is mainly effective on Gram-negative bacteria. It exerts its antimicrobial effects by causing bacterial cytoplasm membrane depolarization, increas- ing cell membrane permeability and disturbing the membrane integrity of Gram-negative bacte- ria. MDAP-2 may offer a new strategy to for defense against Gram-negative bacteria.
One of the methods of sewage sludge disposal, which is based on its fertilizing properties, is its use in nature, e.g. in farming (if all the permissible standards are met). However, the sludge used for soil fertilization might also contain heavy metals, pathogenic microorganisms, thus causing contamination in soil foundation and deterioration of the conditions for development of indigenous organisms. Particular threat is posed by the existence of drug-resistant microorganisms in sewage sludge. This problem has not been researched in detail yet. The authors of the present study aimed to determine qualitative changes in drug-resistant microorganisms in sandy soil fertilized with selected sewage sludge. Sewage sludge after different types of drying process (natural and solar) was added to the degraded sandy soil. The effect of the methods of sewage sludge drying on concentration of drug-resistant microorganisms in soil fertilized with the sludge was analysed. The study demonstrated that sewage sludge dried naturally in drying beds pose threat to soil environment and, potentially, to people and animals which have contact with fertilized soils. In sandy soils fertilized with these types of sewage sludge, pathogenic forms which exhibit resistance to first-line antibiotics can be found.
Due to insufﬁcient operation efﬁciency, the studied treatment plant has undergone modernization. The aim of this study was to assess whether this modernization improved quality of the STP efﬂuent and water quality in the receiver. The research period of ﬁfty months covered time before and after the modernization. Samples were collected in four sites – upstream and downstream of the STP and by the sewage discharge. Electrolytic conductivity, water temperature and pH were measured onsite. Chemical analyzes were based on ion chromatography and determined the concentration of NH4+, NO3-, NO2-, PO43-, TDS. Microbiological analysis comprised serial dilutions to assess the number of mesophilic and psychrophilic bacteria and membrane ﬁltration to enumerate E. faecalis, total and fecal coliforms as well as total and fecal E. coli. Values of most analyzed parameters did not improve after the modernization, or improved for a very short period of time (NH4+), while some of them even increased, such as PO43-, total and thermotolerant coliforms and E. coli. The maximum value of thermotolerant E. coli reached nearly 7 million CFU/100 ml and was observed after modernization. Also at the sites situated downstream of the STP some of analyzed parameters increased. The conducted modernization did not improve the quality of treated sewage and even a further deterioration was observed. It could have been a result of rapidly growing number of tourists visiting the studied area, thus generating large amounts of sewage causing STP overload coupled with poor water and wastewater management. Signiﬁcant percentage of unregistered tourists hinders proper assessment of the STP target efﬁciency.
To improve bioremediation of arsenic (As) contamination in soil, the use of microorganisms to efficiently reduce As and their assessment of genetic erosion by DNA damage using genomic template stability (GTS) evaluation and using RAPD markers were investigated. The five sites examined for microorganisms and contaminated soils were collected from affected gold mining areas. The highest As concentration in gold mining soil is 0.72 mg/kg. Microorganism strains isolated from the gold mining soil samples were tested for As removal capacity. Two bacterial isolates were identified by 16S rRNA gene sequence analysis and morphological characteristics as Brevibacillus reuszeri and Rhodococcus sp. The ability to treat As in nutrient agar (NA) at 1,600 mg/L and contaminated soil samples at 0.72 mg/kg was measured at 168 h, revealing more efficient As removal by B. reuszeri than Rhodococcus sp. (96.67% and 94.17%, respectively). Both species have the capacity to remove As, but B. reuszeri shows improved growth compared to the Rhodococcus sp. B. reuszeri might be suitable for adaptation and use in As treatment. The results are in agreement with their genetic erosion values, with B. reuszeri showing very little genetic erosion (12.46%) of culture in As concentrations as high as 1,600 mg/L, whereas 82.54% genetic erosion occurred in the Rhodococcus sp., suggesting that Rhodococcus sp. would not survive at this level of genetic erosion. Therefore, B. reuszeri has a high efficiency and can be used for soil As treatment, as it is capable to tolerate a concentration of 0.72 mg/kg and as high as 1,600 mg/L in NA.
This study characterizes mycorrhiza helper bacteria (MHB) from selected unpolluted locations as well as subjected to industrial emissions. To determine the species of bacteria isolated from the roots of ectomycorrhizal pine and birch, a method based on the sequence analysis of a 16S rRNA gene was used. The isolated bacteria were initially characterized by available biochemical methods and phenotypic observation. On the selected bacteria representatives isolation of DNA was performed, on which the PCR reaction was carried out. In this way amplified samples were automatically sequenced and the obtained results were compared to public databases. Among the isolated bacteria Pseudomonas fluorescens SBW25 and Burkholderia xenovorans LB400 species were dominant.
The aim of this work was to determine the inﬂuence of various variants of bioleaching on effectivity of releasing chosen critical metals: rhodium, cadmium, indium, niobium and chromium from ashes which are a byproduct of municipal waste and sewage sludge thermal processing. The research was conducted in 3 variants that considered different process factors such as temperature (24ºC and 37ºC), mixing intensity and aeration. After 5 days of the process the analyses were made of metals content, sulfate concentration, pH, general number of bacteria number, index of sulfur oxidizing bacteria. The best results of bioleaching were achieved by running the process at the temperature of 24ºC with aeration. The efﬁciency of rhodium and cadmium release from the byproduct of municipal waste thermal processing was above 90%. The efﬁciency of indium and chromium release reached 50–60%. Only niobium leached better in mixing conditions. The byproduct of sewage sludge thermal processing was far less susceptible to bioleaching. The highest effectivity (on a level of 50%) was reached for indium in temperature of 24°C with aeration. The efﬁciency of bioleaching depended on waste’s physiochemical properties and type of metal which will be released. Aeration with compressed air had a positive inﬂuence on the increase of sulfur oxidizing bacteria what corresponded with almost double increase of sulfate concentration in leaching culture. Such conditions had a positive inﬂuence on the increase of the efﬁciency of bioleaching process. Heightening the temperature to 37°C and slowly mixing did not impact bioleaching in a positive way.
Three chromium resistant bacterial strains, Pseudomonas fluorescens PF28, Enterobacter amnigenus EA31 and Enterococcus gallinarum S34 isolated from tannery waste contaminated soil were used in this study. All strains could resist a high concentration of K2Cr2O7 that is up to 300 mg/L. The effect of these strains on clover plants (Trifolium campestre) in the presence of two chromium salts CrCl3 and K2Cr2O7 was studied in soil microcosm. Application of chromium salts adversely affected seed germination, root and shoot length. Bacterial inoculation improved the growth parameters under chromate stress when compared with non inoculated respective controls. There was observed more than 50% reduction of Cr(VI) in inoculated soil microcosms, as compared to the uninoculated soil under the same conditions. The results obtained in this study are significant for the bioremediation of chromate pollution.
One of the prerequisites for sustainable development is integrated waste management, including sewage sludge. Besides its good fertilization properties, sewage sludge, which is an inevitable by-product of sewage treatment, accumulates toxic chemical substances and dangerous pathogenic and toxicogenic organisms. Uncontrolled introduction of sewage sludge into soil might pose a serious threat to food chain and natural soil microflora. This in effect might disturb the ecological balance in a particular ecosystem. This study presents author’s own investigations of the sanitary conditions of sewage sludge and the conditions after the processes of aerobic and anaerobic stabilization. The investigated sewage sludge originated from a municipal wastewater treatment plant. The sewage sludge samples were transferred onto proliferation and diagnostic media. The results of the analysis obtained in this study confirmed that sewage sludge is a material which is rich in microorganisms, including pathogenic bacterial species such as: Escherichia coli and Salmonella typhimurium. Mycological tests demonstrated that sewage sludge is a material which is conducive to proliferation of yeast-like and mould-like fungi, among which both pathogenic and toxinogenic species can be present. Quantitative analysis of the investigated sewage sludge demonstrated that the processes of stabilization reduce the content of microorganisms but they do not guarantee product safety in sanitary terms. A huge variability and variety of biological composition points to the need for further research in the field of sanitary characteristics of sewage sludge and survival rate in microorganisms from different types of sewage sludge.
Root associated bacteria were isolated from Suaeda nudiflora and two isolates were selected for this study: rhizospheric Bacillus megaterium and endophytic Pseudomonas aeruginosa. These isolates were inoculated into maize variety Narmada Moti during its germination. TTC (2, 3, 5-triphenyl tetrazolium chloride) staining was used to confirm the association of the isolates with the maize root. The effects of these root associated bacteria were tested alone and in combinations for cell wall reinforcement and the induction of defense enzymes such as phenylalanine ammonia lyase (PAL) and β-1,3-glucanase in the presence of fungal pathogen Aspergillus niger in maize. The results indicated that the rhizospheric bacteria had a greater fight response to fungal infection than the endophhytic bacteria due to cell wall lignification as well as the rapid induction of higher concentrations of defense related enzymes.
The laboratory experiment was set up on a podzolic soil in two variants. In one of them non-sterile sewage sludge was introduced into the soil, and in the second - the same sludge but subjected previously to the process of sterilisation. In both variants the same doses of the sludge were applied: 30 (1%), 75 (2.5%), 150 (5%), 300 (10%) and 600 Mg·ha-1 (20%). Then, after 0.5, 1, 2, 3, 4 and 5 months, the soil of both experimental variants was analysed for the numbers of bacteria and fungi decomposing proteins, the rate of the process of ammonification, the rate of the process of nitrification, and for proteolytic activity. The results obtained revealed a stimulating effect of the sludge, both sterile and non-sterile, on the numbers of the microbial groups under study and on the rate of nitrification and protease activity. Only the process of ammonification was subject to inhibition. The observed effects of the sludge were the most pronounced in the case of the higher sludge doses. Significantly greater numbers of protein-decomposing fungi and higher activity of almost all (except for ammonifcation) analysed biochemical parameters in the soil with non-sterile sludge compared to that with sterile sludge indicate an effect of microorganisms from the sludge on the microbiological transformations of nitrogen in soil amended with sewage sludge.