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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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Use of anti-stress agents to minimize the consequences of stressors on the production of Horticultural Crops
Nosálková, Monika ; Hnilička, František (advisor) ; Dana, Dana (referee)
The objective of the thesis is to research the physiological differences between picked genotypes of Capsicum annuum and the effect of lower temperatures in the beginning of ontogeny. Another objective is to find out how Capsicum annuum plants react to application of anti-stress agent Atonic and to determine the most appropriate physiological indicators of resistence to low temperatures.
Capsicum annuum ranks among thermophilic crops for which the temperature optimum is of 22 - 25 °C during the day and 18 - 20 °C overnight. They are planted at the field station at the time when ground frost may occur. These can cause physiological changes in the plant or even cause her death.
Three varieties were chosen for the experiment: Amy, Eva a Lydia. Plants were divided into a control group and treatment group. Both groups were further divided into two additional groups - with and without the application of Atonic in the beginning of vegetative growth (6th day of exposure to stress). Control group plants were grown at 20 °C and stressed plants were moved into a klimabox with temperature of 5 °C throughout the entire day. Such temperature may occur after planting the plants into an outdoor habitat.
Proline content in leaves of Capsicum annuum and relative discharge of electrolyte was studied in two - day intervals. From the obtained results it can be concluded that the accumulation of proline was the highest for Lydia variety in a stressed scenario with the application of Atonic and the lowest proline content was found in in the Amy variety in a control group. Furthermore, it was discovered that the ontogeny of plants has an effect on the accumulation of proline and relative discharge of electrolyte from the beginning of the measurement. The absolute highest electrolyte discharge was measured in the Lydia variety in a stressed scenario and the lowest one was present in the Amy variety in a control group. These results confirm the hypothesis.
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The transformation of the short countryside prose in the second half of the 19th century and at the beginning of the 20th century (Hálek, Světlá, Mrštík, Rais, Nováková, Kratochvíl, Šlejhar)
Kletečková, Martina ; Mravcová, Marie (advisor) ; Merhaut, Luboš (referee)
Through the interpretation of the short rural prose of five selected authors of the second half of the nineteenth and the beginning of the twentieth century (K. V. Rais, J. K. Šlejhar, A. a V. Mrštíkové, T. Nováková, J. Kratochvíl) the thesis deals with the topic of the rural family according to a few aspects that were chosen on the basis of the historical and sociological findings of the technical literature about the rural family in the nineteenth century. The aspects treated by the above mentioned authors are ordered into the chapters "The family model", "Home and world", "Family and the rural year", "Family and the conditions of property", "Birth and death in the rural family" and "Family and traditions and the norms of religious life". The works of the authors are inserted into the context of the rural prose of the nineteenth century. The follow-up interpretation proves the differences in the approach of the authors to the depicted topic.
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The most frequent disease of German boxer
Augstenová, Jennifer ; Rozinek, Jiří (advisor) ; Karlasová, Barbora (referee)
This bachelor thesis deals with the most frequent disease of German boxer. The history and description of the boxer breed is shortly described at the beginning. German boxer is very popular type of dog. It is a breed that has been originally used for hunting and fight with wild animals. Later was boxer trained as a police dog. Now it is mostly kept as family companion due to its great personality.
In next part of the thesis, every new chapter begins with anatomy of organ where disease occurs. Then there are described the most frequent diseases of German boxer based on consultations with veterinary and scientific literature. Sympthoms, prevalence, diagnosis and potential treatment are described, mostly with pictures.
Spondylosis deformans which affects German boxer the most, must have X-ray examination. Dilated cardiomyopathy and arrhythmogenic cardiomyopathy of the right ventricle are responsible for the most bereavement of dogs of all the described diseases.
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Glasshouses of Buquoy
Vojtěchová, Blanka ; Bareš, Jiří (referee) ; Čechura, Jaroslav (advisor)
This work concems the history of glassworks and glashouses in the region of South Bohemia - Novohradské hory - on the border with Austria. Glassmakers worked here over 500 years, since Middle Age until the beginning of the 20th century. There were many generations of glassmasters, glassmakers, refiners ant others and others in the manufactures and later in the factories. The period before the half of the 18th century. It was the time of first Middle Age Glasshouses in the 14th and 15th century, the time ofthe renaissance glassworks and first baroque glassworks. There had been an excellent glassworks in Wilhelmberg, near Heilbronn, in the Gratzen domain, founded by the Rosenbergs about 1580s. At that time, glassmakers came to this region maybe from Italy and later sure from France. The most importent figure of The period since half of the 18th century, the period of founding of new glassworks in Novohradské hory. During 40 years 8 glassworks were founded. On the beginning, the most famous period of glassworks began here when the Count Georg Franz Buquoy found new glass material - black, later red, hyalite and took all glassworks in the region under his direkt ownership. There where 6 glassworks in the first half of the 19th century. After death of Georg Franz Buquoy began slowly dec1ine of glassworks In...
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Stalin's monument in Prague
Šindelář, Jan ; Pokorný, Jiří (referee) ; Václavů, Lubor (advisor)
was unveiled on 1st May 1955 as a part of the celebration of 10th anniversary of the end of the WW2; more than two years after Stalin's death. The political situation in the Soviet block was changed after N. S. Khrushchev's speech on the cult of Stalin's personality on the XX congress of the Soviet communist party in February 1956 and also on the XXII congress in autumn 1961. The satellite states had to modify their ideology against the cult of personality; therefore also the Stalin's monument in Prague was removed in autumn 1962. Nowadays, on the monument's place there are only stairs, a promenade and a pedestal with a high metronome by V. K. Novák. Stalin's monument in Prague was destroyed after only 7 years of its existence by the same political power which built it. From the beginning the project was intensively monitored and supported by the most powerful memebers of the leading communist party, but the main political line for Soviet satellites was ordered from Moscow. The "rise and fall" of Stalin's monument in Prague is a good example of this fact.
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