All the nematodes found in body cavities of the examined endemic Baikal fishes: 88 Baikal yellowfin Cottocomephorus grewingkii (Dybowski, 1874) and 35 longfin Baikal sculpin Cottocomephorus inermis (Yakovlev, 1890) were identified as Contracaecum osculatum baicalensis (Mozgovoi and Ryjikov, 1950) L3 larvae. The prevalence, mean intensity, intensity range and abundance of the nematodes in C. grewingkii were 37.5%, 2.55, 1-31, and 0.96, respectively, the corresponding values in C. inermis were 60.0%, 2.43, 1-10, and 1.46. The infestation level in C. grewingkii was significantly higher than in C. inermis (Mann-Whitney U-test, p<0.02). The number of parasites was found to increase with the fish length. Although in both and C. inermis, the anisakids were more frequent in males (prevalence of 52.17 and 67.76%, respectively) than in females (prevalence of 35.39 and 42.86%, respectively), differences between the sexes in the infestation level in the two species were not significant (Mann-Whitney U-test, P= 0.09 and P=0.23, respectively). The molecular method applied (PCF-RFLP) allows to identify all the nematodes in both examined fish species as C. osculatum baicalensis.
Rapid molecular biological methods for prenatal diagnosis of the most common aneuploidies, collectively known as rapid aneuploidy testing, are compared in this review. We discuss methodological problems and limitations of these various methods. All these techniques are believed to be accurate and carry a low risk of misdiagnosis, but they differ in terms of labour-intensity and amenability to automation and high throughput testing. The question how to apply them safely and economically in a clinical setting has not been answered yet. The discussed techniques are so far not used as stand-alone tests, but some of them are routinely applied as a preliminary test that shortens the waiting time for classic cytogenetic karyotyping. In the future, mainly because of economical reasons, these methods may replace cytogenetics in the category of patients who make up the majority of those currently offered prenatal karyotyping: patients with moderately increased risk and no abnormalities detected by ultrasound.
Many ecological terms are very difficult to define, or to apply to some groups of organisms. In the case of fungi, it is unclear how to speak of individuals, populations and species. Relying on morphological characters often gives ambiguous results. However, in recent years new techniques have been developed, techniques that provide us with robust tools, helpful in recognizing specific groups in nature. This paper aims to give a short overview of how molecular methods are used to delimitate species, individuals and populations in fungi.
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.
PL
W prezentowanej pracy skupiono się na porównaniu metod identyfikacji mikroorganizmów, w tym przypadku podobieństwa pomiędzy szczepami, z wykorzystaniem klasycznych metod płytkowo-mikroskopowej i molekularnej. Obie metody zastosowano, opierając się na założeniu potwierdzonym w ostatnim czasie wskazującym na znaczne zróżnicowanie w obrębie rodzaju Trichoderma, utrudniającym dokładną morfologiczną identyfikację. Obecnie kładzie się również silny nacisk na molekularną identyfikację gatunków. Porównanie metod klasycznej i molekularnej oparto na zastosowaniu typowych wyznaczników makro- i mikroskopowych oraz molekularnych RAPD i RTLP-PCR. Rezultaty wskazały, iż żadna z zastosowanych metod nie jest do końca adekwatna, jednak dokładniejsze wyniki uzyskano, stosując metodę wykorzystującą techniki molekularne. Niestety, nie zaobserwowano powiązania pomiędzy RAPD i RFLP, a ich zderzenie wręcz komplikowało interpretację wyników. Eliminacją tych problemów może być zastosowanie innych markerów molekularnych
The aim of the study was to identify and differentiate five strains isolated from fermented dairy products using species-specific polymerase chain reaction (PCR) as well as PCR of internal transcribed spacer (ITS-PCR) and pulsed field gel electrophoresis (PFGE) in reference to type strains. Results of species-specific PCRs showed that three strains belonged to the species of L. helveticus and two strains to the species of L. casei. Results obtained with both IST-PCR and PFGE method showed low diversity of the isolates since only three different ITS-PCR and PFGE profiles were obtained. Moreover, differentiation conducted merely with the PFGE method allowed distinguishing the type L. casei DSMZ20011, L. rhamnosus DSMZ20021 and L. paracasei subsp. paracasei DSMZ5622 strains. Results of this study confirmed that, although time-consuming and expensive, the PFGE method was characterised by the highest discriminatory power in strain differentiation. The ITS-PCR method even though fast, easy and relatively inexpensive, showed to be more suitable for the pre-selection of strains.
Haemophilus parasuis belongs to opportunistic microorganisms of undefined virulence. The purpose of the studies was to compare suitability of PCR/RFLP in our modification and ERIC PCR for epidemiological study of domestic strains of H. parasuis. The results were evaluated taking into account two different aspects: suitability of the tests for isolating the highest possible number of clone groups and subjective evaluation of the method judged with respect to the following criteria: difficulty, availability of equipment and reagents as well as time and cost of the study. The results obtained in the present study show that the two methods used for typing of H. parasuis had high discriminatory power. Taking into account this parameter it can be concluded that ERIC PCR is more suitable than PCR/RFLP. This justifies the use of ERIC PCR for routine epidemiological analyses of mentioned pathogen. Taking into account the complexity of method used, ERIC-PCR based on random amplification of DNA, proved to be comparable to PCR/RFLP. The last mentioned technique is relatively less expensive and labour-consuming, especially when diagnostic PCR method is used for the epidemiological studies.
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