Andrzej Bieganowski

Determination of the electrokinetic potential of Rhizobium leguminosarum bv trifolii Rt24.2 using Laser Doppler Velocimetry — A methodological study

Electrokinetic potential (ζ, zeta potential) is one of the parameters which characterize the physicochemical properties of the bacterial cell envelope. The term is often used in the context of adhesiveness of bacteria and biofilm formation. This work presents the methodological aspects of zeta potential determination in strain Rt24.2 of Rhizobium leguminosarum using Laser Doppler Velocimetry combined with Phase Analysis Light Scattering and changed electric field techniques. The influence of media (0.9% NaCl, 0.2% NaCl, TY, GYM, 79CA, 20E and M1), temperature of measurement, number of measurement repetitions, phase of culture, concentration of bacteria, and storage at low temperature on the value of electrokinetic potential was investigated and a comparison was drawn between live and dead bacteria. All of those factors modified the zeta potential, showing that these parameters should be precisely specified in studies of bacterial electrokinetic potential, which is not always done. The obtained results also indicated that the zeta potential of Rhizobium leguminosarum should be determined directly in samples without storage at a defined bacterial density. The measurement should be done only once in a sample inserted into the cell of a measuring device to eliminate changes occurring in the sample (increase of electrolytic conductivity) under the electric field used.

Influence of the Sand Particle Shape on Particle Size Distribution Measured by Laser Diffraction Method

Abstract The aim of this paper was to show how the shape of sand particles affects the results of particle size distribution obtained by the laser diffraction method. On the basis of the results obtained one can conclude: the shape of the investigated particles influences particle size distribution obtained by the laser diffraction method. This phenomenon occurs in the sand fraction, as shown in our investigation. The importance of this effect depends on the type of the measured material and on the aim of the investigations. For most researchers in soil science and sedimentology who investigate sand fractions, this impact can be negligible. Further investigations with other soil and sediment fractions are needed.

Application of Gas Sensor Arrays in Assessment of Wastewater Purification Effects

A gas sensor array consisting of eight metal oxide semiconductor (MOS) type gas sensors was evaluated for its ability for assessment of the selected wastewater parameters. Municipal wastewater was collected in a wastewater treatment plant (WWTP) in a primary sedimentation tank and was treated in a laboratory-scale sequential batch reactor (SBR). A comparison of the gas sensor array (electronic nose) response to the standard physical-chemical parameters of treated wastewater was performed. To analyze the measurement results, artificial neural networks were used. E-nose—gas sensors array and artificial neural networks proved to be a suitable method for the monitoring of treated wastewater quality. Neural networks used for data validation showed high correlation between the electronic nose readouts and: (I) chemical oxygen demand (COD) (r = 0.988); (II) total suspended solids (TSS) (r = 0.938); (III) turbidity (r = 0.940); (IV) pH (r = 0.554); (V) nitrogen compounds: N-NO3 (r = 0.958), N-NO2 (r = 0.869) and N-NH3 (r = 0.978); (VI) and volatile organic compounds (VOC) (r = 0.987). Good correlation of the abovementioned parameters are observed under stable treatment conditions in a laboratory batch reactor.

Improvement of counting helminth eggs with internal standard.

The sanitary regulations introduce a requirement for parasitological control of sewage water used for irrigation and fertilisation of crops. The existing methods of estimation of helminth eggs, based on the coproscopic method using sedimentation, desorption, centrifugation and flotation of the material, allow for only 30-75% efficiency. The aim of this paper is to present an improved quantitative estimation method. The proposed method is based on an internal standard. A predetermined number of eggs of Ascaris lumbricoides suis L is coloured with crystalline violet. These marked eggs are added to sample being examined. Counting the marked eggs in the sample and comparing them with their introduced number allows to estimate the efficiency of the procedure. This results in an increase in the efficiency of the method to 100%. It is possible to calculate the real number of helminth eggs in the sample on the basis of the estimated efficiency of the procedure. The sensitivity of the proposed method is at the level of about 0.4 ppm. The necessary number of replications in the method is about 10 times lower than that in the existing methods. The number of natural (not coloured) eggs does not influence the efficiency of the method with an internal standard.

Production of exopolysaccharide by Rhizobium leguminosarum bv. trifolii and its role in bacterial attachment and surface properties

Background and aimsThe acidic exopolysaccharide (EPS) produced by Rhizobium leguminosarum bv. trifolii is required for the establishment of effective symbiosis with compatible host plants (Trifolium spp.). In the rhizobium-legume interaction, early stages of root infection and nodule development have been well studied from a genetic standpoint. However, factors important for colonization of several surfaces by rhizobia, including soil particles and roots, have not yet been thoroughly investigated. The aim of this study was establishing of environmental factors affecting production of EPS by R. leguminosarum bv. trifolii strain 24.2 and the role of this polysaccharide in bacterial surface properties and attachment ability.MethodsBesides the wild-type strain, its derivatives differing in the level of EPS produced were used to these analyses. The ability of attachment to abiotic and biotic surfaces of these strains were established using CFU counting experiments. Three-dimensional structure and other parameters of biofilms formed were characterized in confocal laser scanning microscopy. Electrokinetic (zeta) potential of rhizobial cells were determined using Laser Doppler Velocimetry.ResultsIt was evidenced that the ability of R. leguminosarum bv. trifolii to produce EPS significantly affected bacterial attachment and biofilm formation on both abiotic and biotic surfaces. In addition, the presence of this polysaccharide influenced the zeta potential of rhizobial cells. Mutant strains having a mutation in genes involved in EPS synthesis were significantly impaired in attachment, whereas strains overproducing this polysaccharide showed higher adhesion efficiency to all of the tested materials. EPS facilitated attachment of bacterial cells to the tested surfaces most probably due to hydrophobic interactions and heterogeneity of the envelope surface.ConclusionsEPS produced by R. leguminosarum bv. trifolii plays a significant role in attachment and biofilm formation to both abiotic and biotic surfaces as well as bacterial surface properties.

MEASUREMENT OF ACTIVATED SLUDGE PARTICLE DIAMETERS USING LASER DIFFRACTION METHOD

Abstract The paper presents a study on the possibility of using the laser diffraction method for measuring the size of the activated sludge particles. Susceptibility of the particles to mechanical disintegration, dependent on the programmed value of stirring intensity, was observed (stirring was caused by required dynamic flow of analysed suspension through the measurement unit). According to the conclusions presented in this paper, it may be assumed that the laser diffraction method can be applied for measurement of activated sludge particle diameters under the following conditions: 1) the size of activated sludge particles measured by the laser diffraction method is not a real value, but after standardisation of measurement conditions can be treated as a parameter describing the sludge; 2) the particle diameters of activated sludge should be stabilised before the measurement, eg by mixing in the measurement unit or by ultrasound waves application. Abstrakt Praca dotyczy oceny możliwości wykorzystania dyfrakcji laserowej do badania rozmiarów kłaczków osadu czynnego. W czasie pomiarów zaobserwowano podatność analizowanych cząstek na zniszczenie mechaniczne zależną od zaprogramowanej intensywności mieszania (mieszanie i przepompowywanie jest wymagane w celu wymuszenia przepływu analizowanych zawiesin przez układ pomiarowy). Na podstawie wniosków opracowanych na podstawie prowadzonych badań można stwierdzić, iż metoda dyfrakcji laserowej może być stosowana do pomiarów rozmiarów kłaczków osadu czynnego pod następującymi warunkami: rozmiary kłaczków oraz pozostałych elementów osadu czynnego uzyskane w czasie pomiarów nie mogą być traktowane jako wartość bezpośrednia opisująca rozmiary kłaczka, lecz po standaryzacji warunków pomiarowych może być traktowana jako ilościowy parametr opisujący właściwości osadu czynnego. Przed pomiarem metodą dyfrakcji laserowej osad czynny powinien być uprzednio stabilizowany na przykład za pomocą mieszania w układzie pomiarowym bądź też za pomocą ultradźwięków.

Comparison of the dynamics of natural biodegradation of petrol and diesel oil in soil

AbstractContamination of soil with petroleum products is a major environmental problem. Therefore, one of the issues related to environmental protection is assessment of the ability of soil microbial populations to biodegrade petroleum-derived substances. The aim of the study was to compare the dynamics and fractionation of carbon isotopes during biodegradation of selected petroleum products (petrol and diesel) in soil characterised by optimal humidity for plants. The analyses were performed on soil material sampled from the arable layer of a fertile soil (chernozem) in central Poland. The soil samples were treated with two petroleum substances, i.e. unleaded 95-octane petrol and diesel fuel. The dynamics of changes was assessed by monitoring carbon dioxide content and oxygen content in the headspace over the soil surface. Additionally, the ratio of δ13C carbon isotopes was measured in the substrates added (petrol and diesel) and in CO2 emitted to the atmosphere. In summary, it should be concluded that th...

Use of a Dynamic Light Scattering Technique for SDS/Water/Pentanol Studies

The interpretation of micelle/aggregate size obtained by use of the DLS technique for SDS/water/pentanol systems was discussed by comparison of the results of measurement with theoretical data. For most of the studied systems, the apparent radii (R h,app ) did not satisfactorily characterize the size of the aggregates (R h,app < 1 nm). The use of a correction factor (f = 0.26) confirmed that the discrepancies were associated with the electrostatic intermicellar interactions. However, the fuzzy optical interface between dispersed and dispersing phases can also be the reason of such results. An increase of pentanol content caused a decrease of the droplet radius in w/o systems but in o/w systems the changes were negligible.

Surface Properties of Wild-Type Rhizobium leguminosarum bv. trifolii Strain 24.2 and Its Derivatives with Different Extracellular Polysaccharide Content.

Rhizobium leguminosarum bv. trifolii is a soil bacterium able to establish symbiosis with agriculturally important legumes, i.e., clover plants (Trifolium spp.). Cell surface properties of rhizobia play an essential role in their interaction with both biotic and abiotic surfaces. Physicochemical properties of bacterial cells are underpinned by the chemical composition of their envelope surrounding the cells, and depend on various environmental conditions. In this study, we performed a comprehensive characterization of cell surface properties of a wild-type R. leguminosarum bv. trifolii strain 24.2 and its derivatives producing various levels of exopolysaccharide (EPS), namely, pssA mutant Rt5819 deficient in EPS synthesis, rosR mutant Rt2472 producing diminished amounts of this polysaccharide, and two EPS-overproducing strains, Rt24.2(pBA1) and Rt24.2(pBR1), under different growth conditions (medium type, bacterial culture age, cell viability, and pH). We established that EPS plays an essential role in the electrophoretic mobility of rhizobial cells, and that higher amounts of EPS produced resulted in greater negative electrophoretic mobility and higher acidity (lower pKapp,av) of the bacterial cell surface. From the tested strains, the electrophoretic mobility was lowest in EPS-deficient pssA mutant. Moreover, EPS produced by rhizobial strains resulted not only in an increase of negative surface charge but also in increased hydrophobicity of bacterial cell surface. This was determined by measurements of water contact angle, surface free energy, and free energy of bacterial surface–water–bacterial surface interaction. Electrophoretic mobility of the studied strains was also affected by the structure of the bacterial population (i.e., live/dead cell ratio), medium composition (ionic strength and mono- and divalent cation concentrations), and pH.

The first report of the physicochemical structure of chitin isolated from Hermetia illucens.

This is the first report on the physicochemical properties of chitin obtained from larvae and imagoes of black soldier flies (Hermetia illucens). Scanning electron microscopy revealed differences in surface morphologies of the two types of chitin. The crystalline index values of chitins from adult flies and larvae were 24.9% and 35%, respectively. This is a trait that differentiates these biopolymers from chitins extracted from other sources described so far. X-ray diffraction patterns and IR spectroscopy revealed that both types of samples of chitin were in an α crystalline form. Also, the results of elemental analysis, thermal stabilities and FTIR spectroscopy of the chitins from larvae and adults of H. illucens were similar, which points to a general similarity in their physicochemical structure.