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Vliv délky ekvilibrace na motilitu spermií býků po rozmrazení
KRÁTILOVÁ, Karolína
The aim of the study was to monitor the change in motility of bull sperm post-thawing depending on the time of equilibration before freezing of insemination doses. The percentage of progressive sperm activity post-thawing was determined from the average progressive sperm activity after collection. The evaluation was performed on 6 bulls, approximately the same age, reared under the same conditions at the insemination station. 15 ejaculates were taken from each bull and subsequently processed into insemination doses, which were divided into four groups according to the length of equilibration: 1, 3, 8 and 24 hours. Equilibration was performed in a refrigerator at 5 ° C. The doses were then frozen and evaluated after one week of storage in liquid nitrogen. The mean values of the input parameters were: progressive motility at sampling was 74.63 ? 9.07, the amount of ejaculate 10.08 g and the density 0.91x 106 ml-1 sperm. The results document higher progressive sperm motility with longer equilibration times (8 and 24 hours) as opposed to shorter times (1 and 3 hours) (p <0.01). The best procentual progressive motility was recorded at the equilibration time of 8 hours, when it was 58.74 ? 13.92 % of the initial value (43,84 % of real progresive aktivity). When evaluating the progressive motility of bull sperm after thawing, statistical differences were observed between bulls (p <0.05). Determination of the progressive motility of bull sperm is important in terms of assessing bull fertility. The equilibration time is only one of many factors influencing the quality of the produced insemination doses.
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Aggregation of bull seminal plasma protein
Boháček, Hanuš ; Liberda, Jiří (advisor) ; Hýsková, Veronika (referee)
Mammalian fertilization is a sequence of unique and fascinating events, during which seminal proteins are of crucial role. In case of bull (Bos taurus), proteins of seminal plasma (BSP), especially its major component PDC-109, are known to be in aggregated forms, but little is known about mechanism of forming aggregates and their biological function. In present thesis we discovered some interesting properties of PDC-109 and BSP proteins. We found that concentration of these proteins influences their aggregation state significantly, which can be of great biological importance. Separation of seminal proteins by size exclusion chromatography revealed three main fractions denoted I, II and III, with apparent molecular weights of Mr > 150 000, Mr = 30 000 and Mr = 13 000, respectively. In case of PDC-109, molecular weights of theese fractions were retained even after purification procedure, which implies very stable interactions in forming of aggregates. In addition, there was a difference in distribution of PDC-109 glycoforms among fractions, which can be related to the fact, that theese fractions have different sperm membrane binding patterns as we determined by fluorescence microscopy. However, further experiments are needed for better understanding this issue.
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Species-specific structural differences of mammalian sperm and function of their key proteins during fertilization.
Dobrodinská, Anna ; Frolíková, Michaela (advisor) ; Kuntová, Barbora (referee)
The fertilization is a process during which a male and a female gamete merge so that a new organism may come into being. The sperm-egg fusion is preceded by several essential processes, such as the capacitation, acrosome reaction, the sperm binding to the zona pellucida and oolemma, and membrane fusion of the gametes. Numerous proteins, which are located in both sperm and eggs, are major actors in controlling the listed, essential processes. During the process of fertilization these proteins fulfil one or more functions. In mammalian sperm, significant species-specific differences may be found both in their morphology and at the protein level. A complex understanding of species-specific distinctions in sperm structure and functions of key sperm proteins would contribute to a better insight into the process of fertilization, thereby enabling us to better diagnose and subsequently treat the causes of infertility in humans. This bachelor's thesis summarizes the current knowledge of sperm structure and its key proteins that has been acquired through the studies of the following model mammal species: bull, boar, mouse, and human. Further, this thesis brings an interspecific comparison between the studied species. Keywords: sperm, fertilization, acrosome reaction, capacitation, sperm proteins, bull,...
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Species-specific structural differences of mammalian sperm and function of their key proteins during fertilization.
Dobrodinská, Anna ; Frolíková, Michaela (advisor) ; Kuntová, Barbora (referee)
The fertilization is a process during which a male and a female gamete merge so that a new organism may come into being. The sperm-egg fusion is preceded by several essential processes, such as the capacitation, acrosome reaction, the sperm binding to the zona pellucida and oolemma, and membrane fusion of the gametes. Numerous proteins, which are located in both sperm and eggs, are major actors in controlling the listed, essential processes. During the process of fertilization these proteins fulfil one or more functions. In mammalian sperm, significant species-specific differences may be found both in their morphology and at the protein level. A complex understanding of species-specific distinctions in sperm structure and functions of key sperm proteins would contribute to a better insight into the process of fertilization, thereby enabling us to better diagnose and subsequently treat the causes of infertility in humans. This bachelor's thesis summarizes the current knowledge of sperm structure and its key proteins that has been acquired through the studies of the following model mammal species: bull, boar, mouse, and human. Further, this thesis brings an interspecific comparison between the studied species. Keywords: sperm, fertilization, acrosome reaction, capacitation, sperm proteins, bull,...
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Přirozená plemenitba masného skotu na farmách v ČR
Šrubařová, Markéta
The thesis deals with the natural mating of cattle, excluding the market production, with focus on the time period of using a breeding bull and the reasons for their removal from the breeding. The study includes the description of genitalia both male and female, and also the breeding cycle of cows and the ejaculate of bulls. Further on, the study looks into the process of natural breeding, its rules, advantages and disadvantages, and the dates of spring and winter calving. Shortly described is the selection of breeding bulls, their inclusion to the breeding, the time period of their use, the most common reasons for their removal and the options that enable the elimination of those reasons. In conclu-sion, I introduce a real business, which deals with the breeding of cows without market production of milk. The way of breeding is described, and also the factual replacement of bulls in this business, including reasons for their exclusion from breeding.
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Aggregation of bull seminal plasma protein
Boháček, Hanuš ; Liberda, Jiří (advisor) ; Hýsková, Veronika (referee)
Mammalian fertilization is a sequence of unique and fascinating events, during which seminal proteins are of crucial role. In case of bull (Bos taurus), proteins of seminal plasma (BSP), especially its major component PDC-109, are known to be in aggregated forms, but little is known about mechanism of forming aggregates and their biological function. In present thesis we discovered some interesting properties of PDC-109 and BSP proteins. We found that concentration of these proteins influences their aggregation state significantly, which can be of great biological importance. Separation of seminal proteins by size exclusion chromatography revealed three main fractions denoted I, II and III, with apparent molecular weights of Mr > 150 000, Mr = 30 000 and Mr = 13 000, respectively. In case of PDC-109, molecular weights of theese fractions were retained even after purification procedure, which implies very stable interactions in forming of aggregates. In addition, there was a difference in distribution of PDC-109 glycoforms among fractions, which can be related to the fact, that theese fractions have different sperm membrane binding patterns as we determined by fluorescence microscopy. However, further experiments are needed for better understanding this issue.
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Effect of equilibration and freezing parameters on bull sperm motility after thawing
Beránková, Monika ; Stádník, Luděk (advisor) ; Zuzana, Zuzana (referee)
The hypothesis assume the existence differences in bulls spermatozoa viability after thawing, characterized by higher values of motility and higher proportion of live spermatozoa, in case of prolonged equilibration and lower temperature freezing gradient. The aim of this thesis was to find the influence of different equilibration and freezing curve conditions to on spermatozoa viability after thawing. The theoretical part of the thesis summarizes available information about the bulls semen parameters, methods for its evaluation and especially the processing and long-term preservation.
Within the case study part of the thesis, during the years 2013-2015 the semen of the pre-selected group of 5 breeding bulls was being collected at the Sires insemination center. The group was uniform as to age, breed and housing and treatment conditions. After initial assessment the collected semen was divided into parts, which were then processed in different wals. A part of straws was processed by standard protocol, the other part was processed using different length of equilibration and freezing curve.
First part of the straws was equilibrated in a standard protocol for 120 minutes; subsequently half of the equilibrated straws was standardly frozen by the Direct Freezing method based on 3-phase freezing curve while the other half was frozen using the different 2-phase freezing curve. Second part of the straws was initially equilibrated for 240 minutes with the subsequent freezing, similarly as in the previous case, used 3-phase and 2-phase freezing curve. Frozen straws were then stored in liquid nitrogen container at -196 °C.
Spermatozoa motility was evaluated using CASA. The semen was further evaluated on the proportion of live and dead spermatozoa, using coloration of Eosin and Nigrosine, and on the sensitivity of the spermatozoa membranes, using HOS test, which detects the reaction of spermatozoa tail membrane to hypoosmotic conditions.
The obtained results indicated higher progressive spermatozoa motility (+5.57 %) and higher percentage of live spermatozoa (+4.47 %) following the application of the prolonged equilibration (240 min). When applying the length of equilibration 240 minutes, the values of total spermatozoa motility and HOS test were higher as well, though the differences were not statistically significant. When evaluate the effect of using different freezing curves on the parameters of the spermatozoa after thawing, higher values (average + 1.6 %) at almost all of these parameters using 3-phase freezing curve were found, the diferrences were not statistically significant.
Statistically significant differences among the bulls (P < 0.01) were found in total as well as progressive spermatozoa motility, in the proportion of living spermatozoa and in value of the HOS test - the individuality among the bulls is therefore evident. While evaluating the characteristics of spermatozoa movement by CASA, statistically significant differences among the bulls were confirmed at VCL, VSL, VAP, LIN, STR, WOB (P < 0.01), also between the diferrent length of equilibration (P < 0.01), but were not statistically significant when using diferrent freezing curves.
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Factors influencing the bull sperm freezability
Dvorská, Tereza ; Rajmon, Radko (advisor) ; Marie, Marie (referee)
An egg yolk is a common component of diluents used as protectants of sperm cells during the process of cryopreservation. Its substitution by low density lipoproteins (LDL) has shown that it is the LDL that provides the egg yolk with its cryoprotective characteristics: it protects sperms against cold shock and other changes, thus helping to preserve their fertilization ability even after the freezing-thawing process. However, the sperm quality is affected by many other internal and external factors. Therefore, the aim of this study was to evaluate if the effect of the addition of LDL to the diluent of the ejaculate is significantly influenced by the following selected factors - the type of used diluent, the bull´s breed and its individuality and the date of the sampling.
Experimental insemination doses were repetitively (four times) obtained from a group of six bulls (three Holstein bulls and three Czech Fleckvieh bulls) at the Natural Hradišťko insemination service s.r.o. The samples of semen were diluted with two types of non-egg diluents containing soybean lecithin extracts (AndoMed and BioXCell). To each of these diluents, LDL at 4, 6, and 8% concentration was added; a non-LDL diluent served as a check. All the insemination doses were frozen by a standard procedure and then stored in liquid nitrogen. The CASA system (Computer Assisted Sperm Analysis) was selected for the evaluation of the sperm motility. Immediately after thawing and then after two hours of incubation in water bath (37 °C), the values of kinematic parameters were obtained from the samples - the total percentage of motile sperm and the percentage of progressively motile sperm. These data were then statistically processed; based on the outputs, VAP, VCL, ALH and the percentage of progressively motile sperm (PMOT) were chosen as representative kinematic parameters.
The values of the parameters were higher in almost all evaluated samples diluted with BioXCell, compared to those diluted with AndroMed. Even though we demonstrated the existence of a high variability of results depending on the time of incubation, bull breed, the individuality of the bull and the date of the sampling, it could be said that the best concentration of added LDL is 6 % for BioXCell and 8 % for AndroMed.
It would be useful to perform more experiments evaluating the effect of adding LDL to non-egg diluents on the quality of the thawed sperm. In these experiments, more sperm quality parameters should be examined and factors influencing the variability of results demonstrated in this work should be taken into consideration.
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Vyhodnocení kvality jatečně upraveného těla a masa býků českého strakatého plemene skotu s genotypem TT a CT pro leptin
Večerek, Lukáš
The Master's thesis was focused on the estimation of the quality of carcasses and meat of bulls from the Czech fleckvieh cattle with the genotype TT and CT for leptin. The experiment was carried out on 121 slaughter modified bodies. The effect of slaughter age of bulls, the slaughter weight, the "nett" increment and the classifica-tion in the SEUROP system was analyzed. Within the frame of morphometric analysis were provably (p < 0,01) the highest proportions almost in all of the analyzed parts (leg length, leg plenitude, leg circumfer-ence, leg spiral circumference, forequarter length and thorax semi circuit) ascertained by the bulls of the highest age and weight categories (aged 701 -- 822 days or life weight 701 -- 890 kg). A positive correlation was discovered between the class of conformation assessed by the SEUROP methodology and JUT weight when by the deteriorating class of conformation "U" > "R" > "O" was the weight of carcasses (428 > 341 > 294 kg) declining. The lowest content of meat (72,54 +- 2,53 %) with the highest content of separable tallow (10,22 +- 3,24 %) was proved on slaughter bodies of bulls of the highest weight category which is not good especially for the meat processor. The greatest per-cent of dry meat (26,08 %) was provably (p < 0,01) found by the bulls that are most intensively growing, whereas this category of bulls also reaches the highest qualities in the parameter of meat energy value (5797,13 kJ.kg-1). The technologic parameters were provably not influenced by the levels of evaluated factors although some exceptions appeared. The statistically evidential (p < 0,01) differences were noticeable in the size of the area musculus longissimus et thoracis (MLT) when the greatest area MLT (104,24 +- 11,03 cm2) was stated by the bulls slaughtered in the highest age (701 -- 822 days), whereas the strength of muscular fibers was by all three age categories comparable.
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Vlivy působící na tržní produkci jatečných býků a prasat
Oborná, Monika
The aim of the thesis was to evaluate factors influencing the market production of bulls and pigs. For bulls , I focused on the influence of breed , carcass weight and influence of age . In pigs, I watched the effect of carcass weight ( HJUT ) . I started from the measured data that have been gathered under demolition leaves carcasses of bulls and pigs. From these data, I analyzed the impact on bulls breed Holstein cattle , Czech Pied cattle and Holstein crossbreds and Czech Pied cattle. Then I dealt with a market value of pigs and bulls , which I set as the sum of the prices of individual cut-out parts of their carcasses. I assessed the relationship of cutting prices of individual parts of the total sales of carcasses of bulls and pigs. Likewise, I also evaluate the weight of each cut-out portion to the total weight of carcasses of bulls and pigs. Regarding the relationship between výsekovými parts of carcasses of pigs and bulls and total prices for slaughtered uprané body , the highest value of the correlation coefficient in pigs ( r = 0.9958 ) was found in the leg , with the bulls u perk (r = 0.9073 ) .
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