|
|
Ecophysiology of microscopic fungus Pseudogymnoascus destructans
Homutová, Karolína ; Kubátová, Alena (advisor) ; Baldrian, Petr (referee)
A microscopic fungus Pseudogymnoascus destructans (Ascomycota: Pseudeurotiaceae) causes illness known as white-nose syndrome (WNS) causing death of bats during hibernation. The illness occurs in the North America and in Europe. The fungus is characteristic by asymmetrically curved conidia, by slow growth and growth at low temperatures (below 20 řC). The aim of this study is to clarify properties responsible for unique ecelogy of Pseudogymnoascus destructans by comparison with ecological related or unrelated pathogenic or nonpathogenic fungi. This part includes study of tolerance to physiological stresses and recognition of spectrum of utilizating nutrients (compounds of carbon, nitrogen, phosphorus, sulphur and nutrient supplements). Testing to physiological stresses should help to estimate a potentiality of fungus to spread out of caves. The last aim is to develop a selective isolation medium for P. destructans. Influence of several types of physiological stress (e.g. UVA, UVA with UVB, 25 řC, 30 řC, 37 řC and dryness) was investigated with fluorescent stain propidium iodide (PI) by flow cytometry. The spores of Pseudogymnoascus destructans and three fungi from underground spaces were not viable after 3 weeks at 37 řC. Other stresses did not cause a decreasing of viability or some stresses caused...
|
|
|
Diversity and taxonomy of Trichoderma in the Czech Republic
Valinová, Šárka ; Kubátová, Alena (advisor) ; Nováková, Alena (referee)
Genus Trichoderma - anamorph of the genus Hypocrea (Hypocreaceae, Hypocreales, Sordariomycetes, Ascomycota) belongs to saprotrophic filamentous micromycetes. The representatives of this genus are known primarily as cosmopolitan inhabitants of soil ecosystems, including leaf litter and woody debris. This work summarizes the results of the study of isolates belonging to the genus Trichoderma originating from the territory of the Czech Republic, mainly from soil and leaf litter. During the study, altogether 267 isolates were analysed. For all of the isolates the following methods were used: phenotypic characterisation, UP-PCR fingerprinting, gene sequencing in tef 1α and ITS regions, and in the strain AK 115/00 also in rpb2 region. The main result of the study was the identification of 21 species of genus Trichoderma belonging into 4 groups and discovery of one isolate of species yet unknown to science. Species T. crassum, T. gamsii, T. ghanense, T. rossicum and T. longipile were found in the Czech Republic for the first time. According to the literature review and results of this study, 34 different Trichoderma species is currently known from the Czech Republic. In the work there is also discussed low reliability of identification of certain species based only on phenotypic characters. Key words:...
|
|
|
Microscopic fungi in caves and other underground shelters
Horáková, Karolína ; Kubátová, Alena (advisor) ; Hujslová, Martina (referee)
This Bachelor thesis is meant as a literature search concerned with diversity of microscopic fungi in caves, prospectively in underground spaces which were made by humankind. The caves represent unique environment for living organisms which is characteristic with specific microclimatic conditions and with deficient sources of nutrients. I have concentrated my focus to micromycetes which were isolated from sediments, atmosphere, dead insect and excrements in caves. The next part of my Bachelor thesis is aimed at species Geomyces destructans which causes so called white-nose syndrome illness - geomycosis on hibernating bats. It is an actual problem which interests scientists such as mycologists, zoologists and parasitologists.
|
|
|
Decomposition of leaf litter by microscopic fungi in anthropogenic habitats
Bukovská, Petra ; Gryndler, Milan (referee) ; Kubátová, Alena (advisor)
V letech 2006 - 2008 byl na dvou antropogenních stanovištích s výskytem biologické půdní krusty proveden terénní experiment s listovým opadem břízy a modelovou celulózou využívající metodu opadových sáčků. Rychlost dekompozice opadu i celulolytická aktivita byla na obou stanovištích extrémně nízká. Diverzita společenstva mikroskopických hub na opadu, zjišťovaných izolační metodou a metodou inkubace opadu ve vlhkých komůrkách, však byla srovnatelná s diverzitou známou z přirozených lesních ekosystémů temperátního pásma. Také sukcesní změny společenstva hub pozorované na obou lokalitách odpovídaly v základních rysech údajům z jiných typů stanovišť. Metodou ITS-TRFLP kombinovanou s použitím taxonomicky specifických primerů byla odhalena značná diverzita chytridií a naopak jen velmi nízká diverzita bazidiomycetů. Na stanovišti s vyšší mírou antropogenního stresu byla zjištěna nižší diverzita hub, značně odlišná skladba společenstva a nižší dekompoziční aktivita. Na opadu studovaných stanovišť byly pozorovány i druhy, u kterých nebyla dosud známa tolerance k přítomným antropogenním stresům, a druhy dosud nepopsané. Powered by TCPDF (www.tcpdf.org)
|
|
| |
|
|
Diabetes mellitus negatively affects male reproductive parameters in vivo
Valášková, Eliška ; Žatecká, Eva ; Pavlínková, Gabriela ; Bohuslavová, Romana ; Dorosh, Andriy ; Elzeinová, Fatima ; Kubátová, Alena ; Margaryan, Hasmik ; Pěknicová, Jana
According to the World Health Organization (WHO), 15% of couples in reproductive age suffer from infertility problems, and up to 60% of cases are caused by male factor. This could be caused by genetic background, environmental factors and various diseases, including diabetes mellitus (DM). However, the impact of DM on male fertility is not fully understood. . The aim of this study was to investigate the effects of DM on reproductive parameters and sperm quality, using mouse model. DM (type 1) was induced by Streptozotocin in FVB inbred mouse strain. Mice with blood sugar levels higher than 13.9 mmol/L were considered diabetic. After 4 weeks of diabetes exposure, diabetic males were bred with wild type females and transgenerational effect of DM was assessed. Selected morphological, cellular, and molecular parameters of diabetic males and their male offspring were compared to appropriate controls. There was an increased in sperm fragmentation and abnormalities of sperm morphology in diabetic mice in both generations. An increased staining with apoptotic marker annexin V was also detected in the diabetic groups. Furthermore, a presence of protamines as major sperm nuclear proteins was analysed. Protamine 1 to protamine 2 ratio (P1/P2), a marker of male fertility, was altered in sperms of experimental diabetic animals in both generations. Our findings indicate that DM type 1 negatively affects sperm quality and P1/P2 ratio and this negative effect is transmitted to the progeny
|
|
|
Expression of selected proteins in sperm in men with normal and pathological spermiograms using monoclonal antibodies
Pěknicová, Jana ; Čapková, Jana ; Dorosh, Andriy ; Margaryan, Hasmik ; Kubátová, Alena ; Děd, Lukáš
Recent studies show that infertility in human populations it affects an estimated 15% of couples of reproductive age. Male infertility is the primary cause for 60% of these cases. For these reasons, we analyzed the acrosomal and sperm surface proteins in men with normal and pathological spermiograms. We found that intra-acrosome proteins: TERA (Transitional endoplasmic reticulum ATPase), GAPDHS (Sperm Glyceraldehyde-3-phosphate dehydrogenase), and PRKAR2A (C-AMP-dependent protein kinase II, PRKAR2A), which can be identified using our monoclonal antibodies, are different express in healthy men and men with astenozoospermia (with reduced sperm motility), and with a significantly reduced expression in the astenozoospermia. These proteins are involved in energy metabolism and apoptosis of the cells, and some of them in the sperm-egg interaction; therefore, they have an important role in reproduction. On the other hand, there were no statistically significant differences in the expression of surface proteins (Appolipoprotein J (Clusterin) and Semenogelin). Our findings show that astenozoospermia as a complex disorder of the semen is often combined with other pathological conditions that are not diagnosed by the semen analysis. Therefore monoclonal antibodies are so suitable instrument for the detection of proteins associated with the pathology of the sperm in the semen with low sperm motility. In general, monoclonal antibodies against the sperm proteins are an appropriate tool to detect sperm quality in reproductive medicine.
|
|
| |
|
|
Fluorescent analysis of the differential protein expression in normozoospermic and asthenozoospermic sperm samples
Děd, Lukáš ; Čapková, Jana ; Kubátová, Alena ; Teplá, O. ; Pěknicová, Jana
Asthenozoospermia is one of the main seminal pathologies underlying male infertility. Previous proteomic studies have demonstrated the significant differences in the protein profiles between normozoospermic and asthenozoospermic sperm samples. Since these studies were primarily focused on the identification of differentially expressed proteins by mass spectrometry, we aimed to evaluate the ability of our diagnostic antibodies to detect the differential expression of selected protein markers by fluorescent microscopy and flow cytometry techniques. Therefore, we analyzed sperm samples from 30 men with normal and 30 men with astheno spermiograms, average by the panel of our diagnostic anti-human sperm (Hs) antibodies. Fluorescent microscopy and flow cytometry analysis revealed quantitative differences in the protein abundances between normo and astheno sperm samples, namely, in GAPDHs, evaluated with Hs-8 MoAb, VCP, evaluated with Hs-14 MoAb, and ATP synthase, evaluated with MoAb Hs-36. From the methodological point of view, we observed very high correlation between the data obtained by fluorescent microscopy and flow cytometry techniques and therefore both methods are useful for evaluation of protein differences associated with asthenozoospermia. From the clinical point of view, we observed the strong association of the low sperm motility in the sample with the expression of proteins, playing an important role in sperm energy metabolism (expected), but also with the expression of all tested intra-acrosomal proteins. These findings further demonstrate asthenozoospermia as a complex semen disorder frequently associated with other semen pathologies, which are not diagnosed by basic semen analysis, and the possibility to use monoclonal antibodies as a tool for diagnosis of protein associated sperm pathologies in the semen with the low sperm motility.
|
|
| |
guest ::
RSS feed.