Present work contains the attempt to compare methods of microorganisms identification, for example, the similarity between species – molecular and classical. The both methods were used because classical approaches based on the use of morphological criteria are, as in several other fungi, difficult to apply in Trichoderma, due to plasticity of characters to interpret. Consequently, almost all recent studies have used molecular data to characterize and identify species. The comparison of these two techniques is based on using guidelines of macroscopic and microscopic evaluation of few classical factors as well as RAPD and RFLP – molecular techniques. Results indicate that none of the technique being compared is adequate when used alone, although molecular ones are more accurate. There is no complete convergence between RAPD and RFLP what makes it even harder to interpret but this fact can be the result of inconvenient markers used in experiment.
Ovoalbumine preparation obtained as by-product during isolation of lysozyme and cystatin from egg white was used as substrate for proteolytic enzymes. The protein was degraded by plant serine proteinase isolated from Cucurbita ficifolia and by two yeast proteinases: aspartic and serine isolated from Yarrowia lipolytica. Degradation process was monitored by determination of the hydrolysis degree and the increase of free amino groups concentration. The obtained hydrolysates were also fractioned by RP-HPLC. It was shown that the highest degree of hydrolysis (45%) of ovoalbumin preparation was achieved with plant proteinase. Peptide profiles of the protein hydrolysates confirmed differences in specificity of tested proteinases against ovoalbumin preparation.
The assessment of the microbiological quality of golka – a smoked regional cheeses produced from unpasteurized mixed milk (sheep and cow) in the Carpathian Mountains region as well as a primary identification of LAB derived from these products were the main objectives of this study. The counts of aerobic bacteria, LAB, S. aureus, yeasts and molds was determined. The presence of Salmonella and Listeria was also investigated. The cell counts of microorganisms was determined using standard plating techniques, and the presence of selected indicator strains was established using the ELFA method. The qualitative analysis of the LAB population was performed using PCR-based analysis of metagenomic DNA isolated from natural golka microbiota. Dominating LAB species were further identified using PCR-RFLP and 16S rRNA sequencing. The results of the performed studies have revealed, that the cell counts of microorganisms inhabiting golka cheeses was similar regardless the fact, that the samples were provided by different suppliers. The population of acidophilic LAB was the most abundant, reaching the level of 6.9x109 cfu٠g-1. Salmonella, Listeria, enterotoxic S. aureus strains and molds were not detected in the analyzed cheese samples. Nevertheless, all golka samples showed presence of yeasts ranging from 1.2x106 to 1.6x107 cfu٠g-1. LAB from the genera Enterococcus, Lactococcus, Lactobacillus and Leuconostoc were detected in all samples. Lb. casei strains were dominant among 166 tested isolates..
