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Freezing technology of bull sperm in relation to its survivability and fertilization ability
Doležalová, Martina ; Stádník, Luděk (advisor) ; Jiří, Jiří (referee)
The aim of optimalization the insemination doses production is to provide the highest fertilization ability of spermatozoa during the demanding proces of processing fresh semen and its subsequent cryopreservation. Temperature changes causes spermatozoa damage during the cooling and freezing. Spermatozoa is exposed to cold shock and many others limiting factors, which leads to cell death and therefore to decline of fertilization ability of thawed insemination doses. For increasing spermatozoa resistance, exactly the plasma membrane resistance against cold shock was fraction of egg yolk LDL cholesterol (low density lipoprotein) at various concentrations into the comercially produced diluents added. It is believed that LDL acts possitively to plasma membrane and helps to maintain the fertilization ability of spermatozoa after thawing. Following step in the proces of insemination doses production is slow cooling of diluted semen and equilibration, when the straws are store at cooling box for 30 minutes to 240 hours. This period is necessary to penetrate of certain diluent components into the spermatazoa also maintain the balance between their intracellular and extracellular concentration. Also important is subsequent freezing temperature gradient of insemination doses. The most suitable freezing method is based on computer controlled temperature decline in freezing chamber which allows the precise control of ice crystals formation that could tear and kill the cell.
During 2012 to 2016 was repeatedly collected semen from the group of breeding bulls (n = 27, Holstein and Czech Fleckvieh breed) at AI centre. Semen which fulfill the standard entrance conditions in first step was evenly into several parts divided. For dilution the three types of comercially diluents AndroMed, Bioxcell and Triladyl with and without LDL addition were used. Into the diluents AndroMed and Bioxcell the concentration of LDL 4 %, 6 % and 8% into the dilent Triladyl 6 %, 8 % and 10 % was added. Diluted semen was filled into the glass capillares with volume 0,1 ml and temperature +4 °C. Subsequently the sample was placed to cold bath (0°C) for 10 minutes. Then the volume of capillare with physiological solution (37 °C) was mixed and for next 120 minutes was incubate. The effect of cold shock to proportion of live spermatozoa was evaluated by using Eosin and Nigrosine staining technique during heat test of spermatozoa survivability after spermatozoa heating and after 120 minutes of incubation.
The more suitable semen diluents which provide the higher spermatozoa resistance against cold shock were AndroMed and Bioxcell. Together the possitive effect of LDL addition into the diluents to lower decrease of proportion of live spermatozoa during heat test was found (P<0.05). The most suitable LDL concentration which had a favorable influence at spermatozoa resistance against cold shock was 6 % in diluent Bioxcell. Values of the proportion of live sperm were higher at the beginning of the heat test (+1.31% to + 3.2%) and after 120 minute incubation (+5.82% to +8.41%) compared to other diluents with and without addition of LDL.
In the next step the process of equilibration was optimized, is an important part of insemination doses production. The effect of the length of equilibration for subsequent fertilization ability of spermatozoa was evaluated using spermatozoa motility based of CASA and proportion of live spermatozoa after thawing and during heat survival test lasting 120 minutes (37 ° C). Suitable semen was diluted by comercially used diluent AndroMed based on soya lecithin, filled into the straws (0.25 ml), cooled and equilibrated in cooling box for 30, 120 and 240 minutes and freezed in programmable freezing box applying four types of freezing curves differing in temperature rate decline. There was used standard and by producer recommended 3. phase freezing curve, then 2. phase freezing curve, and 3. phase freezing curve with slower as well as rapid decline of temperature rate in freezing chamber, compared with standard freezing curve. The highest spermatozoa motility was found using 240 minutes of equilibration by +2.72% and +4.58% compared to other lengths of equilibration (P <0.05 to 0.01). The highest proportion of live spermatozoa was found using 120 minutes of equilibration (+6.87 % and +8.68 %). The highest average spermatozoa motility during heat test after thawing was achieved by using 2. phase freezing curve (from +2.97% to +10.37%, P <0.05), also in the proportion of live spermatozoa (from + 4.37% to +8.82%, P <0.01). When evaluating interaction between the length of equilibration and freezing curve (standard 3. phase and 2 . phase freezing curve), the highest average spermatozoa motility and proportion of live spermatozoa using 240 minutes of equilibration by both freezing curves was reached, there was no statistically significant differences. As well as, in all evaluated parts of this study the individual differences between ejaculate of bulls and within semen from one bull (P <0.05) as secondary effect were found.
To maintain good fertilization ability of semen during cryopreservation is necessary to increase the spermatozoa resistance against cold shock using addition of correct concentration of LDL into the commercially used diluents AndroMed and Bioxcell. Subsequently the fertilization ability of insemination dose is influenced by cooling, the length of equilibration and freezing. The length of equilibration 120 minutes and more as well as gentle way of freezing according to freezing curve, which ensures a gradual decrease of temperature in freezing chamber provided the higher average spermatozoa motility and proportion of live spermatozoa.
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Individual human odor as a forensic trail in criminal proceeding made by contact or contactless transmission and its resistance to physical agents
Santariová, Milena ; Bartoš, Luděk (advisor) ; Chmelíková, Eva (referee)
The dissertation thesis consists of four papers that have been published in scientific journals.
Study n. 1
The need to recover evidence from water is quite common in criminal investigation. The article deals with the possibility to collect human scent from such objects and with the ability of specially trained dogs to match such scent samples with scent samples collected from detained suspects. During an experiment, designed as a blind one, it has been proved, that human scent can survive on submerged objects and can be later used for scent identification.
Study n. 2
To collect odors the Czech Republic Police use special fabric sorbents manufactured under the registered mark ARATEX. Before use the fabric sorbent is treated by water vapor sterilization. After the scent identification the sorbent is destroyed. The goal of the study was to verify if the vapor sterilization is effective enough to remove human scent that has already penetrated into the sorbent structure or in other words if the sorbent can be exposed to vapor sterilization and then used again. Specially trained dogs were used to match starting odors with target odors in the line-ups. The results showed that dogs are able to correctly match human odors even after they have being exposed to vapor sterilization.
Study n. 3
The purpose of the study was to determine the temperature at which the human scent is degraded so that a dog would not be able to identify it. In contrast to expectations, eight dogs used in the experiment almost flawlessly identified human scents from five scent donors exposed to temperatures of 100°C, 200°C, 300°C, 400°C, 500°C, 600°C, 700°C, and 800°C. Only two of the dogs were able to identify 5 of 15 scent samples exposed to 900°C. No dog identified a scent exposed to 1000°C. Our study verified heat survivability of human scent far beyond existing expectations.
Study n. 4
We tested the hypothesis that if odor fallout (the release of a human odor onto an untouched object) in human subjects exists, then holding a hand above an absorbent will produce a detectable scent which will be subsequently matched in a detection test by trained dogs. Scents were collected from seven males to sterile cotton absorbent squares. The left hand was used to get the control scent and the right hand served as the target scent. Each experimental subject was sitting and his left hand was laid down on a cotton square for 3 min. The right hand was held 5 cm above another cotton square for 3 min. The scent identification was done by two specially trained police German shepherds. Both dogs performed 14 line-ups and correctly matched the collected scents of all test subjects. The results suggest the existence of human odor fallout, whereby a human scent trace is left by humans even if they do not touch an object.
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Total contents and speciation of arsenic and selenium in plants growing in soils with different physico-chemical properties
Tremlová, Jana ; Száková, Jiřina (advisor) ; Vymazal, Jan (referee)
This dissertation thesis deals with uptake, accumulation and transformation of arsenic and selenium in plants growing on soils with different chemico-physical soil properties. The contents of arsenic were investigated in 99 wild plant species, coming from 27 different families and 6 different vegetables growing in arsenic contaminated soils. The results suggest that there are species-specific mechanisms which protects certain plants from the excessive uptake of arsenic from the soil. On the other hand, there are plants using a strategy of accumulation, which theoretically may increase the risk of arsenic entry into the food chain, whether through grazing cattle and wildlife or through consumption of vegetables growing on arsenic contaminated soils. The most prevalent arsenic compounds are AsV and AsIII. Other arsenic compounds occure as minor species. An important finding was detection of arsenobetaine in Plantago lanceolata L. and Carex praecox Schreb ..
The contents of selenium were examined in 73 species of wild plants, coming from 29 different families. The results suggest that naturally low levels of selenium in the soil of selected locations along with a low capability of selenium uptake via collected plant species lead to the low content of selenium in plant biomass which may contribute to selenium deficient throughout the food chain. Foliar application of selenate on wild plant communities and on some types of vegetables in our case Brassica oleracea var. italica can have a positive impact on increasing the selenium content in the aboveground biomass of these plants and by extension, increase the selenium content in the human diet. Dominant selenium compounds in the aboveground biomass of the analyzed plants were SeVI and selenomethionine, with variable proportion of other commonly occurring organic selenium compounds, which is mainly affected by plant species.
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Non-invasive methods for sex and steroid homones determination in gibbons of the genus Nomascus
Bolechová, Petra ; Bartoš, Luděk (advisor) ; Pavel, Pavel (referee)
The gibbon primates of the family Nomascus are classified as critically endangered species, and, to date, basic understanding and information about their biology is missing. With regard to the status of these animals in the wild and captive populations in zoos, being familiar with their reproduction, may improve captive breeding programs. Data collection in this study was to be carried out by the practical use of non-invasive methods (polymerase chain reaction for DNA extraction and enzyme immunoassays), using faecal samples for analysis. The first method, polymerase chain reaction, was used for sex determination in juveniles, because of their coat colour and visual similarity of secondary sex characteristic in both sexes; it is not possible to determinate sex without handling the animal. Another main purpose of this study is to try and answer the hypothesis regarding the ovarian cycle of females, factors influencing their hormone concentration and also the onset of sexual maturity in females and the timing of their fur colour change. Hypotheses were checked by monitoring the concentration of progesterone and oestrogen faecal metabolites and by evaluation of the composition of breeding groups of gibbons with the ZOO influence. During a four year period (from 2010 till 2014), there were a total of 51 animals analysed from 16 different zoos with a faecal sample count totalling 1618 samples. The results confirmed the use of noninvasive methods for sex determination, and thus ensuring the maximum welfare standards. Endocrinological analysis confirmed the hypothesis of the influence of the environment (ZOO) to the hormone concentrations and female´s pregnancy effect in conjunction with her age on the final results of the male - father hormone concentrations. The initial information in this study is the confirmation of the onset of ovarian cycle in young females without connection to their fur colour change and a significant factor of a mother´s impact and her territoriality. This study is the first to present a long term monitoring of ovarian cycles in females and hormone concentrations of other individuals, both male and female, and from various age groups. The results allow us to understand the possible impact of zoo environments on the reproductive status of these gibbons and contribute to the general improvement of breeding management.
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Collective invesment
Černý, Štěpán ; Veselá, Kamila (advisor) ; Jitka, Jitka (referee)
The main object of this Diploma Thesis is according to collective investment through mutual funds formulate appropriate portfolio model for different types investors of retail banking. Theoretical part of the thesis is focuses on information and rules of investements, advantages and disadvanteges of collective investmentm fund types and their strategies. The practical part parses the selected mutual funds and then compares according to historical performance and riskiness. In the last part of it is handled by the main target. In this section are created model portfolia for individual investors, which are diversified and selected investment companies. Each of the suggestions and recommendations of the model portfolios correspond to the principles of conservative, balanced and dynamic investor.
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The collection of published articles
DVOŘÁK, Petr
The aim of the collection of published articles is to define the requisite social skills of the English language teacher as a part of his/her professional competence, in the context of communicative language teaching in classroom interactions between English teachers and adolescent pupils. The theoretical preliminaries of the research whose findings are presented in the articles are the functional communicative approach to language and communicative language teaching. The attention is paid to the processes of interaction, communication and classroom discourse at school with an emphasis on foreign language teaching. We try to specify social skills of the English teacher and define social-skill aspects of foreign language educational interactions. The findings of the research presented in the articles concern mainly the educational style of English teachers and English teacher's behaviour in particular classroom interactions. The research is also focused on selected social-skill aspects of classroom discourse within communicative language teaching, specifically, pupils' and teachers' initiations of communication, questions, and the distribution of communication opportunities.
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Are There Any Digital Curators in Czech Libraries?
Římanová, Radka ; Melichar, Marek
The development of digital repositories reveals the need for new Czech terminologies for library positions. One such job title or specialization is “digital curator”, a term which refers to a specialist responsible for digital collections. Competencies for digital curators include technical, organizational, communication, and marketing skills as well as some level of expertise in computer science as well as in information and library science. A survey of several administrators of Czech digital libraries asks how to best professionally label this group of library staff with respect to the actual activities they perform in their jobs - digital curators, administrators of digital libraries, or digital librarians? Job titles have a strong impact on higher education curricula, on the management of libraries, and also on the coordinated system of remuneration of employees at libraries, academic, and research institutions.
Fulltext: idr-1041_3 -  PDF; idr-1041_4 - PDF
Slides: idr-1041_1 - PDF; idr-1041_2 - PDF
Video: idr-1041_5 -  MP4
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Methodology of the Cooperation between the Director and Actor and Contermporary Theater Practice
Glogrová, Kateřina ; BURIAN, Jan (advisor) ; HRBEK, Daniel (referee)
The thesis deals with the cooperation between the director and actor during individual phases of the staging process as they appear in everyday theatre life. The indiviual phases of the staging process are supplemented by the author's own experience with managing the actors, which she gained especially in the course of her studies at the Drama Faculty of AMU. The second part of the thesis investigates the issues of inner truthfulness and the possibilities of its achievement within the production of a play. As a part of collecting the material for the thesis, the author conducted a research, which included interviews with three professional directors - Hana Burešová, Ladislav Smoček and Štěpán Pácl, whose methods of work are described. In the final part of the thesis the author states that it is difficult to draw general conclusions about the cooperation between the director and actor because it is always of subjective nature. The issue of inner truthfulness is similarly difficult to generalise. Due to the multiplicity of options, however, neither of them loses attractiveness for further research.
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Corporate Culture and Its Change
VONDROVÁ, Martina
The opening part of the bachelor´s work is focused on the literary research, specifically, it introduces some definitions of corporate culture, its components and strength. Furthermore it focuses on describing ways of creating and changing corporate culture. The practical part contains qualitative research on corporate culture in a specific company. The data for the research were collected by means of interviews, observation and intra-office documentation. The conclusion of the work contains findings with further suggestions for improving the corporate culture of the company.
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