Fluorescence microscopy for studying the viability of micro-organisms in natural whey starters

AIMS: The aim of this work was to study the viability and cultivability of microbial populations of different natural whey starters and to evaluate their resistance to thermal treatments (such as exposure to high or low temperatures). METHODS AND RESULTS: Twenty-three natural whey starters for Grana Padano cheese were investigated and subsequently pH measurement, plate count agar using Man-Rogasa-Sharpe (MRS) pH 5.4 agar and whey agar medium (WAM) were performed using these samples. LIVE/DEAD BacLight bacterial viability kit was used. Total count and viability of all the 23 samples were high and similar to each other (CV 20%). However, the cultivable population was lower in terms of cfu ml(-1) and number of cells per millilitre than the viable fraction and highly variable, although its count value was higher in WAM than in MRS pH 5.4. The heating (60 degrees C for 5 min and 54 degrees C for 1 h) and freezing (-20 and -80 degrees C) treatments affected the cultivability and viability of the microbial population. CONCLUSIONS: This study demonstrated the effectiveness of LIVE/DEAD BacLight bacterial viability kit, which has already been used to evaluate bacterial populations, in investigating microbial viability in a complex ecosystem such as a natural whey starter. Significance and Impact of the Study: The aim of this study was to quantify the presence of damaged nonviable bacterial cells in natural whey starters. The Thoma Glass is a useful method to obtain fluorescence microscopy counts to evaluate the technological performance of natural whey starters.     

Applications of a new optical technique for measuring the dielectrophoretic behaviour of micro-organisms

It is shown that the dielectrophoretic behaviour (motion in non-uniform a.c. electric fields) of micro-organisms can conveniently and reproducibly be measured by monitoring the decrease in optical absorbance of a cell suspension as the cells are collected at a micro-electrode array. The dielectrophoretic behaviour, as a function of the frequency of the applied electric field and conductivity of the supporting solution, can be determined more quantitatively and rapidly than by methods so far described in the literature. Results are presented for Micrococcus lysodeikticus, Bacillus subtilis and Escherichia coli for the frequency range 20 Hz to 4 MHz and theoretical considerations are presented for the effect of solution conductivity. A value of 0.2 S/m has been derived for the effective conductivity of M. lysodeikticus.     

Tribo-electrochemical technique for studying tribocorrosion behavior of biomaterials

Tribocorrosion is the term which describes the synergy between tribological and electrochemical processes. An apparatus was designed and built to study the tribocorrosion behavior of biomaterials. Electrochemical set-up with three electrodes is used for controlling the potential of the surface of a conducting material subjected to classical wear testing. Using this equipment, it is possible to carry out friction and wear tests in electrolytic solution under well-defined electrochemical conditions determined by the applied electrode potential. In this paper, this apparatus was described and the tests of deposited TiN on pure Ti for corrosion and tribocorrosion behavior under simulated body fluid were conducted. The presence of TiN layer on the surface of Ti has increased the open circuit potential. The charge transfer resistance (R(ct)) determined using electrochemical impedance spectroscopy (EIS) was higher for the nitrided surfaces than for the Ti substrates. However after wear test, R(ct) was significantly reduced because the protective layer was damaged.     

Application of the lumped age-class technique to studying the dynamics of malaria-mosquito-human interactions

Background

There is an increase of serum levels of IgE during Plasmodium falciparum infections in individuals living in endemic areas. These IgEs either protect against malaria or increase malaria pathogenesis. To get an insight into the exact role played by IgE in the outcome of P. falciparum infection, total IgE levels and functional anti-parasite IgE response were studied in children and adults, from two different endemic areas Gabon and India, exhibiting either uncomplicated malaria, severe non cerebral malaria or cerebral malaria, in comparison with control individuals.

Methodology and results

Blood samples were collected from controls and P. falciparum -infected patients before treatment on the day of hospitalization (day 0) in India and, in addition, on days 7 and 30 after treatment in Gabon. Total IgE levels were determined by ELISA and functional P. falciparum -specific IgE were estimated using a mast cell line RBL-2H3 transfected with a human Fcε RI α-chain that triggers degranulation upon human IgE cross-linking. Mann Whitney and Kruskall Wallis tests were used to compare groups and the Spearman test was used for correlations. Total IgE levels were confirmed to increase upon infection and differ with level of transmission and age but were not directly related to the disease phenotype. All studied groups exhibited functional parasite-specific IgEs able to induce mast cell degranulation in vitro in the presence of P. falciparum antigens. Plasma IgE levels correlated with those of IL-10 in uncomplicated malaria patients from Gabon. In Indian patients, plasma IFN-γ, TNF and IL-10 levels were significantly correlated with IgE concentrations in all groups.

Conclusion

Circulating levels of total IgE do not appear to correlate with protection or pathology, or with anti-inflammatory cytokine pattern bias during malaria. On the contrary, the P. falciparum -specific IgE response seems to contribute to the control of parasites, since functional activity was higher in asymptomatic and uncomplicated malaria patients than in severe or cerebral malaria groups.     

A special technique for studying haptoral sclerites of monogeneans

The marginal hooks and anchors have been digested from the haptors of the monogeneans Gyrodactylus salaris and Discocotyle sagittata. These structures then can be measured and drawn more accurately than those surrounded by haptoral tissue. The technique also facilitates the study of isolated hooks with SEM. The results of studies using this digestion technique are presented.     

A glass fiber filter technique for studying nutrient uptake by fungi: The technique used on colonies grown on nutrient gradients of carbon and phosphorus

In most natural environments supporting fungal growth, nutrients are heterogeneously distributed in space. Growth of a fungus will thus take place in an environment characterized by gradients. A system has been developed for growth of fungi on opposing carbon and mineral nutrient gradients, present in liquid medium in glass fiber filters. By labeling the carbon or phosphorus in the medium, the amount of carbon or phosphorus accumulated inside hyphae ofRhizopus nigricans and the amount still present outside the hyphae were determined. The distribution of labeled C and P in the medium and in the colonies of the fungus grown in the presence and absence of initial gradients in the medium was compared. In both cases, little carbon or phosphorus was found remaining in the medium after fungal growth. With colonies grown on media without an initial gradient, two peaks of carbon and phosphorus accumulation were found, but when there was a gradient there was only one such peak. These peaks coincided with the regions of greatest sporulation. It was concluded, by comparing the distribution of the total amount of carbon inside and outside mycelium grown on gradients with that in control media which was uninoculated, that the translocation of carbon inside the mycelium could have been brought about by simple diffusion.     

Bacterial adherence to hydrocarbons: a useful technique for studying cell surface hydrophobicity

Abstract Bacterial adherence to hydrocarbons (BATH) is a simple and rapid technique for determining cell-surface hydrophobicity. During recent years, this method has found application in the study of the surface characteristics of a wide variety of bacteria and bacterial mixtures. Correlations have been found between the adherence of bacteria to hydrocarbons and their attachment to other surfaces, including non-wettable plastics, epithelial cells, and teeth. A slight modification of the assay enables the isolation of nonhydrophobic mutants. The present publication briefly describes the technique and its modifications, summarizes results obtained using this method, and suggests several directions for further investigation.     

研究基因转录的RT-PCR技术

RT-PCR是检测和定量特异性mRNA的高度灵敏技术,被广泛应用于基因转录水平的分析研究,本文探讨RT-PCR方法的发展,应用及潜在问题。     

A simple method for xylanase preparation used for the hydrolysis and fermentation of hemicellulose to butanediol

Summary When culture filtrates ofTrichoderma harzianum E58 were concentrated by passage through an ultrafiltration membrane with a molecular weight exclusion limit of 10,000 Daltons, 80% of the original xylanase activity was recovered in the ultrafiltrates. Culture filtrates and ultrafiltrates which were concentrated by rotary evaporation contained inhibitors which restricted the fermentation of the hemicellulose-derived sugars to 2,3-butanediol. A simple solvent-exchange treatment of the ultrafiltrates could effectively concentrate the xylanases as well as remove the fermentation inhibitors.