Effectiveness of Mycobacterium paratuberculosis isolation from raw milk by means of direct isolation of DNA and classic culture.

Detection of Mycobacterium paratuberculosis (MAP) in tissues of patients suffering from Crohn's disease has given rise to speculation that this mycobacterium may play some role in the development of this disease in humans. Food products, especially milk obtained from animals infected with paratuberculosis, may be a potential vector of MAP to humans, yet the detection of this pathogen poses a number of difficulties. This study was aimed at comparing the effectiveness of MAP isolation from milk samples. Mycobacteria were detected by means of two methods: direct isolation of DNA using a QIAamp DNA Mini Kit by Qiagen, and a culture method with the use of HEYM culture medium. Analyses were carried out on 87 samples of udder cow milk originating from a herd that exhibited seropositive and serodoubtful reactions against paratuberculosis. The presence of an insertion sequence IS-900 was detected in 18 samples of udder milk analyzed with the method of direct DNA isolation and in two samples analyzed by means of the culture method.

Flow cytometry as a method for the evaluation of raw material, product and process in the dairy industry.

Producing dairy products which are safe for consumers requires the constant monitoring of the microbiological quality of raw material, the production process itself and the end product. Traditional methods, still a "gold standard", require a specialized laboratory working on recognized and validated methods. Obtaining results is time- and labor-consuming and do not allow rapid evaluation. Hence, there is a need for a rapid, precise method enabling the real-time monitoring of microbiological quality, and flow cytometry serves this function well. It is based on labeling cells suspended in a solution with fluorescent dyes and pumping them into a measurement zone where they are exposed to a precisely focused laser beam. This paper is aimed at presenting the possibilities of applying flow cytometry in the dairy industry.

Ca(2+) -dependent plant response to Pb(2+) is regulated by LCT1.

Tobacco plants transformed with TaLCT1 were cultured on Knop's medium with modified calcium concentrations (0.01-3 mM) in the presence of Pb(2+), and in soil contaminated by lead. A 4-5 microM Pb(2+) administered in the presence of 1 mM Ca(2+) inhibited the root growth of transgenic plants to much lesser degree than of control plants, whereas in the presence of 3mM Ca(2+) no differences were found between the studied lines. The reduction of Pb(2+) toxicity in the presence of 1 mM Ca(2+) was not accompanied by a change in the lead tissue concentration. However, when Ca(2+) level in the medium was lowered to 0.01 mM, several fold higher root/shoot Pb ratio in transgenic plants was observed, twofold increase in the total amount of metal accumulated, and lower concentration of Pb in the xylem sap. Results suggest the involvement of TaLCT1 in the regulation of Ca-dependent Pb-detoxification, and under conditions of low calcium in lead uptake and distribution.