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The Polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate (SDS-PAGE) has been used for qualitative and quantitative analyses of milk proteins. Electrophoretically pure casein fractions and whey proteins were used as standards. The binding of Coomassie brilliant blue R-250 by the particular proteins was found to differ. The dye-absorption coefficients calculated to correct the errors of densitometric measurements for αs1-, αs2-, ß-, and k- casein dye complexes were as follows: 1.00; 1.00; 1.07; 0.59, and 1.00; 0.67; 1.34 for α-lactalbumin, ß-lactoglobulin and bovine serum albumin, respectively. The procedure described for the elimination of dye-absorption errors seems more universal and simple for protein quantitation.
Pasteurella multocida (Pm) and Bordetella bronchiseptica (Bbr) are important pathogens of humans and many species of animals. At present, several classic and modern techniques are applied for their detection and identification. The review presents an overview of the diagnostic methods that are the most widely used in laboratories, with special emphasis placed on highlighting their advantages and limitations. The choice of the technique depends on the aim of the study. The most often used are classical methods such as microbiological, serological, biochemical, and - in the case of atrophic rhinitis - likewise the morphometric analysis of the cross-section of a turbinated bone. The above-mentioned techniques are not accurate enough because most of the test results must be confirmed by more precise and sensitive techniques. The development of a wide spectrum of molecular techniques has facilitated the undoubted identification of Pm and Bbr. The widest used is PCR basing on the amplification of the gene encoding dermonecrotoxin of Pm and Bbr. Other molecular techniques could be useful for describing the connection among individual components of bacterial cells and their ability to develop the disease. Moreover they could also be used to describe the degree of phylogenetical relationship among field strains. Such information could be used to make a prognosis regarding the appearance of diseases on a particular area. Taking into account the significance of the rapid and precise detection of the pathological agents for immediate and accurate therapy, the continuation of studies aimed at the developing of rapid and sensitive diagnostic techniques is fully recommended and justified.
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