A new piezoelectric response model for population growth of bacteria

A piezoelectric response model on the population growth of microorganism is proposed. This model is based on a novel population growth model, which has a more obvious ecological meaning and the fact that the series piezoelectric quartz crystal (SPQC) sensor responses to conductivity changes of the medium during the growth of the microorganism. From the response model four parameters can be obtained including the maximum specific growth rate mu(m), saturated population size N(m), and two constants C and K(1). The influence of the parameters on the response curve is discussed in which the influences of mu(m) and N(m) are more obvious. With the proposed model the quantitative determination of bacteria may be more accurate than the frequency detection time (FDT) method. Then the growth of Escherichia coliform (E. coli) monitored with a SPQC sensor is compared with the simulated growth curve obtained by the proposed model and a good agreement is obtained.     

A Rapid Method for Determination of Growth of Thiobacillus Ferrooxidans by Series Piezoelectric Quartz Crystal

ABSTRACT

A rapid and simple method for monitoring the growth of Thiobacillus ferrooxidans is put forward by using a series piezoelectric quartz crystal (SPQC). The SPQC was applied to continuously determine the variation of frequency shift during the culture of Thiobacillus ferrooxidans, with a conductive electrode as the probe. The frequency shift as a function of time agrees with a typical 'S - shape' model, a piezoelectric sensor responding well to the growth of Thiobacillus ferrooxidans. The fundamentals for determination of the bacterial growth activity using the SPQC is also described; the frequency shift is only dependent on the electrical conductivity of the solution and the dielectric constant.     

Effects of low-frequency magnetic fields on bacteria Escherichia coli.

The effects of low-frequency magnetic fields (Bm=2.7-10 mT, f=50 Hz, time of exposure t=0-12 min, laboratory temperature) on the viability and oxidoreductive activity of gram-negative bacteria Escherichia coli were investigated. The growth of these bacteria was negatively affected by such fields. We compared two experimental systems--solenoid [Sb. Lek. 99 (1998) 455] and a cylindrical spool--to find differences between nonhomogeneous and "more homogeneous" magnetic fields. We observed analogous effects in both experimental conditions. The growth curve of the exposed bacteria was lower than the control one. The ability of bacteria to form colonies decreased with increasing magnetic field intensity and with increasing time of exposure. The oxidoreductive activity was measured using reduction of a tetrazolium salt. The decrease in oxidoreductive activity with increasing time of exposure was observed, but the effect was due to a lower amount of bacteria surviving the exposure to the magnetic fields. The decrease in oxidoreductive activity and ability to form colonies were compared with the assumption that the effect of magnetic field is probably bactericidal.     

Effects of low-frequency magnetic fields on the viability of yeast Saccharomyces cerevisiae

A 50 Hz magnetic field effect on the growth of yeasts Saccharomyces cerevisae was studied. The cylindrical coil induced magnetic fields with inductions up to 10 mT. Duration of exposure varied up to 24 min. Exposure took place at laboratory temperature (24-26 degrees C) and the air ventilator maintained the temperature at the place of the sample. We measured the growth curves of yeasts in broth and we calculated the number of CFU (colony forming units) on solid soil. We found that magnetic field decreases the number of yeasts, and slowed down their growth. The result is similar to the experiments with bacteria E. coli, S. aureus and L. adecarboxylata. It seems that the magnetic fields kill a part of yeasts and the bigger part of them survives and continues in their growth.     

Comparison of the low-frequency magnetic field effects on bacteria Escherichia coli, Leclercia adecarboxylata and Staphylococcus aureus

This work studies biological effects of low-frequency electromagnetic fields. We have exposed three different bacterial strains-Escherichia coli, Leclercia adecarboxylata and Staphylococcus aureus to the magnetic field (t<30 min, B(m)=10 mT, f=50 Hz) in order to compare their viability (number of colony-forming units (CFU)). We have measured the dependence of CFU on time of exposure and on the value of the magnetic field induction B(m). Viability decreases with longer exposure time and/or higher induction B(m) for all strains, but the quantity of the effect is strain-dependent. The highest decrease of the viability and the biggest magnetic field effect was observed with E. coli. The smallest magnetic field effect appears for S. aureus. From the measurement of the growth dynamics we have concluded that the decrease of the CFU starts immediately after the magnetic field was switched on.     

Mössbauer, electron paramagnetic resonance, and theoretical study of a high-spin, four-coordinate Fe(II) diketiminate complex

The iron(II) complex LFeCl 2Li(THF) 2 (L = beta-diketiminate), 1, has been studied with variable-temperature, variable-field Mossbauer spectroscopy and parallel mode electron paramagnetic resonance (EPR) spectroscopy in both solution and the solid state. In zero applied field the 4.2 K Mossbauer spectrum exhibits an isomer shift delta = 0.90 mm/s and quadrupole splitting Delta E Q = 2.4 mm/s, values that are typical for the high-spin ( S = 2) state anticipated for the iron in 1. Spectra recorded in applied magnetic fields yield an anisotropic magnetic hyperfine tensor with A x = +2.3 (+ 1.0) T, A y = A z = -21.5 T ( solution) and a nearly axial zero-field splitting of the spin quintet with D = D x approximately -14 cm (-1) and rhombicity E/ D approximately 0.1. The small, positive value for A x results from the presence of residual orbital angular momentum along x. The EPR analysis gives g x approximately 2.4 (and g y approximately g z approximately 2.0) and reveals a split " M S = +/- 2" ground doublet with a gap distributed around Delta = 0.42 cm (-1). The Mossbauer spectra of 1 show unusual features that arise from the presence of orientation-dependent relaxation and a distribution in the magnetic hyperfine field along x. The origin of the distribution has been analyzed using crystal field theory. The analysis indicates that the distribution in the magnetic hyperfine field originates from a narrow distribution, sigma phi approximately 0.5 degrees , in torsion angle phi between the FeN 2 and FeCl 2 planes, arising from minute inhomogeneities in the molecular environments.     

Rapid detection of Escherichia coliform with a bulk acoustic wave sensor based on the gelation of Tachypleus amebocyte lysate

A new sensing method (BAW-TAL technique), which combined the bulk acoustic wave (BAW) technique with the gelation reaction of Tachypleus amebocyte lysate (TAL), was used for viscosity and density measurement and applied to the detection of Escherichia coliform (E. coli). This method depended on the fact that the viscosity and density of the mixture increased, and as a result, the resonance frequency decreased correspondingly after TAL was mixed with the heated E. coli solution that contained endotoxin. Results showed that the frequency shift was linearly related to the logarithm of E. coli concentration in the range of 2.7x10(4)-2.7x10(8) cells/ml. The correlation coefficient was 0.996. This BAW-TAL method was compared with the standard pour plate counts (PPC) method. The proposed method was much more rapid and simpler for detection of E. coli than the traditional methods.     

Insulator-based dielectrophoresis for the selective concentration and separation of live bacteria in water

Insulator-based dielectrophoresis (iDEP) was utilized to separate and concentrate selectively mixtures of two species of live bacteria simultaneously. Four species of bacteria were studied: the Gram-negative Escherichia coli and the Gram-positive Bacillus subtilis, B. cereus, and B. megaterium. Under an applied direct current (DC) electric field all the bacterial species exhibited negative dielectrophoretic behavior. The dielectrophoretic separations were carried out in a glass microchannel containing an array of insulating posts. The insulating posts in the microchannel produced nonuniformities in the electric field applied along the channel. Mixtures of two species of bacteria were introduced into the microchannel and the electric field was applied. The bacterial species exhibited different dielectrophoretic mobilities under the influence of the nonuniform field. From these experiments a trapping order was established with E. coli trapping at the weakest applied electric field, while the Bacillus species were trapped at different characteristic threshold fields. At stronger applied electric fields, the two different species of bacteria in the microchannel were dielectrophoretically trapped into two spatially distinct bands. The results showed that iDEP has the potential to selectively concentrate and separate different species of bacteria.     

Escherichia coli O157:H7 detection limit of millimeter-sized PZT cantilever sensors is 700 cells/mL.

A composite self-excited millimeter-sized lead zirconate titanate (PZT) glass cantilever (2 mm x 1.8 mm; sensing area of 6 mm2) was fabricated for the detection of Escherichia coli (E. coli) O157:H7. The fundamental and second mode resonance in air was 10.95 +/- 0.05 kHz and 43.45 +/- 0.05 kHz, respectively. Affinity purified monoclonal antibody (anti-E. coli O157:H7) specific to the pathogen E. coli O157:H7 was immobilized at the cantilever glass tip, and then immersed in liquid containing the pathogen (70 to 7 x 10(7) cells/mL). The resonant frequency showed a reduction and reached a steady state shift of 0 +/- 5, 46 +/- 5, 260 +/- 5, and 1010 +/- 5 Hz corresponding to 0, 700, 7000, and 7 x 10(7) cells/mL. From the experiments conducted, the detection limit of the sensor was 700 cells/mL.     

An electrochemical method for simultaneous detection and identification of Escherichia coli, Staphylococcus aureus and Salmonella choleraesuis using a glucose oxidase-peroxidase composite biosensor

Quantification of bacterial pollution by amperometric detection at 0.0 V of glucose consumption at a graphite-Teflon-glucose oxidase-peroxidase-ferrocene composite biosensor under flow injection conditions is reported. Using Escherichia coli as the model bacterium, the composition of the growing medium was optimized. A constant glucose concentration of 4.0 x 10(-4) M was added to the culture medium. The relative response to glucose, expressed as the ratio between the amperometric signal and the signal at incubation time t = 0 multiplied by 100, as a function of E. coli concentration, showed a typical behaviour. Limits of detection of 6.5 x 10(2) or 6.5 cfu mL(-1) were achieved after 3 or 7 h of incubation, respectively, with no pre-concentration step. The detection of bacteria did not affect the lifetime of the biosensor. The feasibility of the detection of Staphylococcus aureus and Salmonella choleraesuis throughout the glucose consumption measurement at the composite biosensor is also demonstrated. The capability of bacterial identification by evaluation of bacterial growth in the culture medium containing the antibiotics polymyxin B, vancomycin, erythromycin, bacitracin, chloramphenicol, tetracycline and ciprofloxacin, was investigated. Each micro-organism tested exhibited a different antibiotic susceptibility profile, thus suggesting the possibility of bacteria differentiation. A rapid methodology for screening of bacteria is proposed.     

Synthesis of DnaK and GroEL in Escherichia coli cells exposed to different magnetic field signals

The effects of extremely low frequency magnetic field (ELF-MF)(1 mT, 50 Hz) on the heat shock protein (HSP) synthesis in Escherichia coli were investigated. Two magnetic field signals were studied: sinusoidal (SMF) and pulsed square wave (PMF). It was found that bacteria exposed to SMF showed a significantly higher level of DnaK and GroEL proteins as compared to sham-exposed bacteria as revealed by Western blot, whereas a lower level was observed after PMF exposure. Similar results were obtained when bacterial cells were exposed to heat shock (HS) after ELF-MF exposure: again SMF and PMF resulted in an increase and in a reduction of HSP amount in comparison with sham control, respectively. In conclusion, the MF influences the synthesis of HSPs in E. coli in a way that critically depends on the signal characteristics.     

A Novel Method for Mutagenicity Test of Carcinogen by using a Series Piezoelectric Quartz Crystal Bactrial Growth Sensor

ABSTRACT

A new method for mutagenicity testing of carcinogens is proposed in this paper. Mutagenic activity is examined through monitoring of the growth situation of Salmonella typhimurium TA98 strain with a series piezoelectric quartz crystal (SPQC) sensor. This method needs no immobilization and preincubation of microorganism, and is simple and rapid. It can be applied to detect mutagen directly. When it was used to study the mutagenicity of dimethyl sulfate, the test time was 4 h at 37°C. There was a good linear relationship between frequency shift and the dose of mutagen in the range 2.5-20 μg/mL and the regression equation was ∠F = 53.82 + 3.81C. Some experimental conditions are also discussed in detail.     

A Series Piezoelectric Quartz Crystal Microbial Sensing Technique Used for Biochemical Oxygen Demand Assay in Environmental Monitoring

A new method for biochemical oxygen demand (BOD) determination, which combines the series piezoelectric quartz crystal (SPQC) technique with the growth of a microorganism is presented in this paper. This method needs no immobilization of bacteria and is simple and convenient. When a calibration technique was used for BOD analysis, the detection time was 2.5 h at 37°C. There was a good linear relationship between the frequency shift and BOD value in the range 2.2–11 mg/L and the regression equation was ΔF = 64.10 + 11.23[BOD]. The proposed method was compared with the conventional BOD₅ method. This method was more rapid than the BOD₅ method and the results obtained by the former were in good agreement with those obtained by the latter. The experimental conditions are also discussed in detail.     

Fourier decompositions and pulse sequence design algorithms for nuclear magnetic resonance in inhomogeneous fields

In this paper, we introduce algorithms based on Fourier synthesis for designing phase compensating rf pulse sequences for high-resolution nuclear magnetic resonance (NMR) spectroscopy in an inhomogeneous B0 field. We show that using radio frequency pulses and time varying linear gradients in three dimensions, it is possible to impart the transverse magnetization of spins phase, which is a desired function of the spatial (x,y,z) location. Such a sequence can be used to precompensate the phase that will be acquired by spins at different spatial locations due to inhomogeneous magnetic fields. With this precompensation, the chemical shift information of the spins can be reliably extracted and high resolution NMR spectrum can be obtained.     

Novel Reduction Kinetics Model of Dimethoate (DMA) on the Growth of Pseudomonas aeruginosa

Abstract

A new method for monitoring Pseudomonas aeruginosa growth with dimethoate (DMA) as sole carbon using the series piezoelectric quartz crystal (SPQC) and high-performance liquid chromatography (HPLC) was described. Compared with the general methods, SPQC gives real-time, multidimensional information. The determination limit of HPLC was 1.08 ng. A combined novel response model for Pseudomonas aeruginosa degrading DMA was derived. By fitting DMA reducing data, the value of the reduction rate constant (k) was obtained and in line with the actual trend. The proposed kinetic model was successfully applied to Pseudomonas aeruginosa degrading DMA.     

Simultaneous detection of Escherichia coli O157:H7 and Salmonella Typhimurium using quantum dots as fluorescence labels

In this study, we explored the use of semiconductor quantum dots (QDs) as fluorescence labels in immunoassays for simultaneous detection of two species of foodborne pathogenic bacteria, Escherichia coli O157:H7 and Salmonella Typhimurium. QDs with different sizes can be excited with a single wavelength of light, resulting in different emission peaks that can be measured simultaneously. Highly fluorescent semiconductor quantum dots with different emission wavelengths (525 nm and 705 nm) were conjugated to anti-E. coli O157 and anti-Salmonella antibodies, respectively. Target bacteria were separated from samples by using specific antibody coated magnetic beads. The bead-cell complexes reacted with QD-antibody conjugates to form bead-cell-QD complexes. Fluorescent microscopic images of QD labeled E. coli and Salmonella cells demonstrated that QD-antibody conjugates could evenly and completely attach to the surface of bacterial cells, indicating that the conjugated QD molecules still retain their effective fluorescence, while the conjugated antibody molecules remain active and are able to recognize their specific target bacteria in a complex mixture. The intensities of fluorescence emission peaks at 525 nm and 705 nm of the final complexes were measured for quantitative detection of E. coli O157:H7 and S. Typhimurium simultaneously. The fluorescence intensity (FI) as a function of cell number (N) was found for Salmonella and E. coli, respectively. The regression models can be expressed as: FI = 60.6 log N- 250.9 with R(2) = 0.97 for S. Typhimurium, and FI = 77.8 log N- 245.2 with R(2) = 0.91 for E. coli O157:H7 in the range of cell numbers from 10(4) to 10(7) cfu ml(-1). The detection limit of this method was 10(4) cfu ml(-1). The detection could be completed within 2 hours. The principle of this method could be extended to detect multiple species of bacteria (3-4 species) simultaneously, depending on the availability of each type of QD-antibody conjugates with a unique emission peak and the antibody coated magnetic beads specific to each species of bacteria.     

INACTIVATION OF GRAM-NEGATIVE BACTERIA BY PHOTOSENSITIZED PORPHYRINS

Photosensitization of Escherichia coli and Pseudomonas aeruginosa cells by deuteroporphyrin (DP) is shown to be possible in the presence of the polycationic agent polymyxin nonapeptide (PMNP). Previous studies established complete resistance of Gram-negative bacteria to the photodynamic effects of porphyrins. The present results show that combined treatment of E. coli or P. aeruginosa cultures with DP and PMNP inhibit cell growth and viability. No antibacterial activity of PMNP alone could be demonstrated and cell viability remained unchanged. Spectroscopically, PMNP was found to bind DP, a mechanism which probably assists its penetration into the cell's membranes. Insertion of DP into the cells was monitored by the characteristic fluorescence band of bound DP at 622 nm. Binding times were 5-40 min and the extent of binding increased with decreasing the pH from 8.5 to 6.5. DP binding constants, as well as the concentrations of PMNP which were required for maximal effect on the various Gram-negative bacteria, were determined fluorometrically. By the treatment of DP, PMNP and light the growth of E. coli and P. aeruginosa cultures was stopped and the viability of the culture was dramatically reduced. Within 60 min of treatment the survival fraction of E. coli culture was 9 x 10(-6) and that of P. aeruginosa was 5.2 x 10(-4). Electron microscopy depicted ultrastructural alterations in the Gram-negative cells treated by DP and PMNP. The completion of cell division was inhibited and the chromosomal domain was altered markedly.     

Change in broth culture is associated with significant suppression of Escherichia coli death under high magnetic field

When Escherichia coli B was cultivated under an inhomogeneous magnetic field of 5.2-6.1 T, a significant 100,000-fold suppression of cell death was observed [Bioelectrochemistry 53 (2001) 149]. The limited magnetic field exposure for 12 h after logarithmic growth phase was sufficient to observe similar suppressive effects on cell death [Bioelectrochemistry 54 (2001) 101]. These results suggest some possible changes in either the medium or the cells during the magnetic field exposure. When the cell-free filtrate of the broth cultured under the magnetic field for 10 h and the cells of E. coli cultivated under the geomagnetic field for 30 h were mixed, and the mixture was subsequently cultivated under the geomagnetic field, the number of cells observed in the filtrate exposed to the high magnetic field was 20,000 times higher than that in the filtrate exposed to the geomagnetic field. When the cells cultivated under the magnetic field for 10 h and the cell-free filtrate of the broth culture exposed to the geomagnetic field were mixed, only a 50-fold difference in the number of cell between under the magnetic field and under the geomagnetic field was observed. This suggests that the filtrate of the broth culture exposed to the magnetic field is primarily responsible for the cell death suppression. It was also revealed that the small difference in pH of the filtrates of the broth culture between under the magnetic field and under the geomagnetic field was critical for the cell death suppression.     

Study on the influence of potassium iodate on the metabolism of Escherichia coli by intrinsic fluorescence

Intrinsic fluorescence, in particular, has the advantage over the extrinsic fluorescence of an unperturbed environment during investigation, especially in complex systems such as biological cells and tissues. Potassium iodate may restrain bacteria growth as well as it acts as an additive in the salt. The influence of potassium iodate (KIO3) on the metabolism of Escherichia coli (E. coli) is investigated for the first time with the intrinsic fluorescence of tryptophan (Trp) and reduced nicotinamide adenine dinucleotide (NADH). We found that potassium iodate may restrain the growth of E. coli as a bacteriostatic agent. When the potassium iodate concentration was below 1.32 mmol/L, the intensity of tryptophan fluorescence decreased linearly whereas the NADH fluorescence did not change. When the KIO3 concentration was over 1.32 mmol/L, the fluorescence of tryptophan and NADH increased a little and their fluorescence intensity decreased when KIO3 was over 6.67 mmol/L. And the bacteria could not continue growing if the KIO3 was over 6.67 mmol/L.We could conclude that potassium iodate has great inhibiting effects on the growth of E. coli through the pathway of protein synthesis and respiratory chain.