The study aimed at optimization of DNA isolation from blood of representatives of four microbial groups causing sepsis, i.e., Gram negative: Escherichia coli, Gram positive: Staphylococcus aureus, yeast: Candida albicans, and filamentous fungus: Aspergillus fumigatus. Additionally, the five commercial kits for microbial DNA isolation from the blood were tested. The developed procedure of DNA isolation consisted of three consecutive steps, i.e., mechanical disruption, chemical lysis, and thermal lysis. Afterward, DNA was isolated from the previously prepared samples (erythrocyte lysis) with the use of five commercial kits for DNA isolation. They were compared paying heed to detection limit, concentration, DNA purity, and heme concentration in samples. The isolation of DNA without preliminary erythrocyte lysis resulted in far higher heme concentration than when lysis was applied. In the variant with erythrocyte lysis, two of the commercial kits were most effective in purifying the DNA extract from heme. Designed procedure allowed obtaining microbial DNA from all four groups of pathogens under study in the amount sufficient to conduct the rtPCR reaction, which aimed at detecting them in the blood.
We introduce three new estimators of the drift parameter of a fractional Ornstein–Uhlenbeck process. These estimators are based on modifications of the least-squares procedure utilizing the explicit formula for the process and covariance structure of a fractional Brownian motion. We demonstrate their advantageous properties in the setting of discrete-time observations with fixed mesh size, where they outperform the existing estimators. Numerical experiments by Monte Carlo simulations are conducted to confirm and illustrate theoretical findings. New estimation techniques can improve calibration of models in the form of linear stochastic differential equations driven by a fractional Brownian motion, which are used in diverse fields such as biology, neuroscience, finance and many others.
Contamination of water environments with herbicides is a common problem nowadays. The aim of the current study was to evaluate the effects of Roundup on common carp ( Cyprinus carpio) after 1, 3 and 10 days of exposure. The used concentrations corresponded to 1 and 5 mg/l of
the active ingredient (glyphosate potassium salt). The haematological analysis performed showed a decrease of the RBC count, as well as an increase of the other erythrocyte indices (Hb, MCV, MCH, MCHC). Most of these changes were dependent on the concentration and time. An increase of the
WBC count and the percentage of immature neutrophils occurred, thus indicating the presence of inflammation. In the studied blood biochemical parameters, only minor and temporary changes were observed. The histopathological analysis revealed no alterations in the gills, liver and kidney. Thus,
the results of the present study suggest that the haematological parameters are more sensitive and reliable markers of the common carp's exposure to Roundup than the other parameters that were tested.
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