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Potential of cholesterol-loaded cyclodextrins and low density lipoprotein for increasing the efficiency of cryopreservation of epididymal sperms in stallions
Janošíková, Martina ; Rajmon, Radko (advisor)
In the case of sudden death or injury stallion reproductive system or necessary castration preventing the collection of semen can to preserve the genetic potential of exceptional individuals take advantage of epididymal sperm. Virtually the only way, how to preserve fertilizing sperm POTENTIAL for long time IS cryopreservation. However, this process induces irreversible changes that are the cause of the reduced number of sperm capable of fertilization after thawing. Epididymal sperm cells have properties different from the ejaculated, harder to keep. Therefore, they are constantly looking for ways to streamline their cryopreservation.
GENERAL most commonly used cryoprotectants include glycerol and Egg yolk. Despite the positive effects is glycerol toxic to cells and Egg yolk also contains ingredients that act on sperm metabolism negatively. Increasing the efficiency of cryopreservation protocols themselves likely to be achieved by replacing toxic components while leveraging industry-produced active yolk fractions.
Submitted dissertation topic itself will deal with the use of cholesterol-loaded cyclodextrins Along with low density lipoproteins, the cryopreservation of epididymal sperm.
Main method: HPLC, Western blot, electrophoresis, fluorescent labels, computer-assisted sperm analysis (CASA).
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Renal denervation of sympaticus in pig
Michálková, Tereza ; Rajmon, Radko (advisor) ; Dita, Dita (referee)
The bachelor thesis deals with methods of renal denervation, a minimal invasive procedure, which is due to interruption of the sympathetic fibers using radiofrequency energy. The consequence is a reduction in blood pressure. The theoretical part deals with the regulation of blood pressure and renal function in regulating the blood pressure. Practical part is focuses on individual made current expert studies and their results.
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Tumor proliferation of mammary gland in bitches
Musilová, Lucie ; Rajmon, Radko (advisor) ; Dita, Dita (referee)
Thanks to breeders' and veterinary care, dogs' age is increasing, which results in proportional increase in diseases, including cancer. Mammary gland cancer is the most common oncologic problem in bitches.
Development of neoplasms is affected by endogenous factors (for example hormones), and exogenous factors also (radiation, carcinogenic substances, etc.). These factors cause mutations, which affects the formation of cancer growth.
We divide tumours on true and false ones. False tumours are e.g. cysts. True tumours are either benign or malignant, which create daughter centres called metastasis. Malignant tumours bear worse prognosis than benign tumours. Metastasis and tumours affect inidividual's organism by e.g. negative effect on organ function. Individual can stimulate tumours by e.g. hormone secretion.
Tumour classification is important because of determining the right treatment and prognosis for the future. Classification is divided in three parts: typing, grading, staging. Typing divides tumours into groups by their effect on tissues, or by individual tumours. Grading divides tumour by level of their differentiation. Staging classifies tumours by TNM, where T defines tumour's size, N defines level of lymphatics invasion and M defines metastasis formation.
Mamary gland function is divided in three physiological phases: mammary gland development, lactation and galactopoesis. Mammary gland is probably modified sweat gland. Lactation is affected by hormone prolactin, which supports mammary gland development. Galactopoesis is state, which maintains lactation. It's important to prevent stress in this phase, because stress hormones block oxytocin, so milk ejection isn't coming about.
Effect of hormones also change by estrous cycle phase. In bitch, there are those phases: proestrus, estrus, diestrus and anestrus. Estrous cycle deffects can occur and those deffects can cause so called false pregnancy. This state can result in diseases like pyometra or mastitis.
Etiopathogenesis of mammary tumours is affected by two kind of factors, endogenous and exogenous. Prevention is also important, this includes more quality care and feed. Another important prevention can be adequate exercise and potential sterilisation of the female. If the animal is already ill, then there is important to discuss it with veterinarian and start treatment as soon as possible. Firt method of treatment is surgical, which is divided by mammary gland invasion. Other method is chemotherapy, which is becoming more and more frequent as support treatment method after surgery. Third method is called radiological. It's relatively new type of therapy, but it is, unfortunately, still relatively expensive. Last method is called hormonal. Again, it is used usually as support treatment after surgical removal of the tumour.
There are used many diagnostic methods for clinical evaluation of mammary gland tumours, e.g. cytology, USG, RTG. Prognosis determination and post-surgical care is also important. Main complication is disease recurrence, which is more frequently occuring in malignant tumours than in benign tumours.
Most important is prevention and treatment start as soon as possible after diagnosis assesment. It is appropriate to not to burden breeding bitch with frequent litters, or let veterinarian perform ovariohysterectomy (OHE) on bitch, which is not used for breeding. Ovariohysterectomy reduces probability of development of this disease.It is very important for breeder to regularly check mammary gland by palpation and immediately visit veterinarian with every change or lump.
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Clinical veterinary diets as a tool for the therapy and management of diseases in dogs
Marešová, Iveta ; Rajmon, Radko (advisor) ; Berka, Pavel (referee)
The aim of the thesis Clinical diets as a tool for therapy and prevention of dog diseases was to elaborate a literary review focused on selected organ and metabolic dog diseases related to nutrition, and a possibility to influence them by clinical diets. Clinical diets are special feed that can be used in case of diseases treatment which can be influenced by the content of a feed ration. It often reduces the need of drugs and in some cases it can significantly extend life of the animal. They also may be used as prevention of serious disease development. On the market there are diets designed for treatments of wide variety of metabolic and organ disorders, e.g. obesity, diabetes mellitus, cardiovascular diseases, gastrointestinal disorders, urinary tract diseases, joint problems etc. Each of the diet has got estimated recommendations which producers should follow to meet nutrients requirements represented in the feed ration and which are specific for the disease. In the cases when a dog was fed an improper ingredients diet or a diet intended for a different type of a disease, it could lead to very serious health damage. Therefore the right and timely diagnosis is very important. Despite of successful treatment the diet is often fed on a long-term or even lifelong basis.
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Inflammation of the mammary gland in mammals
Turečková, Eva ; Rajmon, Radko (advisor) ; Karlasová, Barbora (referee)
This thesis deals with inflammation of mamary gland by mammalia. The work is aimed both at the general level and are capturing species differences in individual mammals, such as livestock and animals hobby farms.
Inflammation of mammary gland, mastitis, is defined as an inflammatory reaction of mammary gland induced when pathogenic microoganisms penetrate to the udder. Introductory part deals with anatomy, development and physiology of mammary gland. The other part is focused on inflammatory reaction generally because the knowledge of this issues is necessary for understanding the processes that are happening in this disease . The last part is focused on the formation, diagnostics, treatment and prevention this disease. The special attention is payed on the prevention in this work because it is the most important key to the success of breeding and to the satisfaction of the animal.
If it will keep proper preventive measures against mastitis with regard to the physiological needs of the animals then we can prevent this disease.
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Silver and its potential use in veterinary medicine
Bušáková, Eliška ; Rajmon, Radko (advisor) ; Berka, Pavel (referee)
Nowadays, inspite of the advanced level of human and veterinary medicine, there are still some areas where available drugs and approaches are not sufficient. It is particulary the increasing resistance of pathogens to used drugs and their side effects that are becoming a serious problem. Having strong antimicrobial effects, silver could be the solution. Its application is not prevalent these days, yet there are many research studies and experiments dealing with its effects on bacteria and viruses, but also with its impact on living organisms. This thesis summarizes and compares them in order to find potential use of silver in veterinary medicine. These studies show that silver may offer a variety of application. However, at first it is necessary to answer the question of potential side effects and to find the right form of silver among various choices that would be the most convenient for the requirements of veterinary medicine.
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Effects of alpha-zearalenol on hyperactivation and acrosomal reaction of the boar sperms
Doleželová, Kateřina ; Rajmon, Radko (advisor) ; Marie, Marie (referee)
Alpha zearalenol is a derivative of the mycotoxin zearalenone, which is commonly found in agricultural crops and commercial feed. It is a secondary metabolite of filamentous fungi of the genus Fusarium. The alpha zearalenol is the result of a biotransformation of zearalenone in the liver and intestinal tissue. When the zearalenone is metobolised this way, it is able to compete for the binding sites of estrogen receptors because the structure is very similar to 17 beta estradiol.
Toxicity of alpha zearalenol therefore lies in an estrogenic stimulation, which induces morphological and functional changes in the reproductive organs and gametes. Toxicity is manifested on the male gamete by reducing viability, motility and the portion of sperms capable of undergoing acrosome reaction.
The aim of this study was to evaluate the effect of alpha zearalenol on the hyperactivation and acrosome reaction of boar sperm depending on the concentration and duration of exposure to mycotoxins.
For the purposes of this study we used two methodologies and the short term chilled commercial insemination doses which were kept in a box with the temperature of 17 degrees of Celsius. The two methodologies had a different count of used concentrations of alpha zearalenol and exposure time. Acrosome reaction of sperms was assessed in both methodologies, while the share of hyperactive sperm was assessed only in the second methodology.
The first methodology did not prove any effect of alpha zearalenol on acrosome reaction, but the second methodology has proven a positive effect of concentrations 5, 10 and 20 uM alpha zearalenol on acrosome reaction of boar sperm. Individual values of parameters from CASA analysis (VCL, ALH, LIN and WOB), which were used for evaluation of hyperactivation of boar sperm, were not affected by any used concentrations of alpha zearalenol.
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Prevention of obesity in dogs
Listíková, Jana ; Rajmon, Radko (advisor) ; Marie, Marie (referee)
Summary
Obesity is a serious condition that brings many health risks and the attendant reduction of the quality of the dog's life. For those reasons, it is important to prevent obesity and to focus on its prevention, which is full responsibility of the owner. The owner should therefore have enough appropriate information about the basic needs of their dog's diet, its natural behavior, and its need to move. The veterinarians play an important role in that .
The aim of this work is to cover the issue of obesity in dogs with the help of research and a questionnaire survey. A part of the aim is to map how owners and breeders cooperate with veterinarians, if/how they are informed about their dog's nutrition, whether they focus on preventing obesity in their dogs, or what they do if their dog is already overweight.
The questionnaires helped evaluate four hypotheses:
1) The dogs whose owners haven't discussed the dog's weight with their veterinarians (while the veterinarians have not offered any consulting about nutrition) suffer from obesity more than others.
2) The dogs whose owners actively focus on obesity prevention suffer from obesity less than others.
3) The dogs who have access to food ad libitum and at the same time their daily activity is less than 1 hour suffer from obesity more than others.
4) The dogs who are fed industrially processed food and whose owners prefer price to quality when choosing food suffer from obesity more than others.
Altogether, 161 questionnaires were filled in and subsequently evaluated. The respondents comprised both males and females of all ages who owned dogs of both sexes and various breeds and ages. This sample of respondents confirmed the first two hypotheses. The other two hypotheses have not been statistically confirmed. Twenty percent of the dogs were categorized as overweight or obese. Overall, the questionnaires showed that in most cases, those respondents trust their veterinarians, they pay attention to prevention and also share the veterinarian's opinion on their dog's condition. On the other hand, the questionnaires also showed that the veterinarians could be more proactive.
Even among the dogs of the above mentioned respondents, there were overweight or obese dogs. Obesity is a hot issue and these days, it is considered a disease. Its prevention should therefore be crucial.
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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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Some factors influencing success of IVF-embryo transfer
Pertoldová, Stanislava ; Rajmon, Radko (advisor) ; Marie, Marie (referee)
Assisted reproduction is a field of medicine that deals with infertility tretment.
Ongoing improvements of laboratory technology as well as ongoing evolving scientific research within IVF research,including experimental embryology in routine practice is contributing in a steady improvement of success rates of assisted reproduction.
This thesis is primarily based on data and information provided by the laboratory of assisted reproduction of GEST IVF Ltd.clinic in Prague,which was collected during the year 2015. This data was analyzed and investigated to verify the effect of selected factors on the success of the treatment. In This thesis we have not used any personal data of patients in accordance with the legislation act.296/2008Sb.
The results show clear evidence between the treatment success rate and patients age within all evaluated parameters. The fundametal hypothesis of interdependence of embryo quality on the further development of the embryo after the transfer was comfirmed.
Conversely, the timeframe for embryo cultivation is not an important parameter of success. Commonly used cuture media also did not show a significant impact on treatment outcome.
Statistical calculations also confirmed surprisingly significant differences in embryo transfer after 72 hours development and 120 hours development for women ever 35 years. Transfers of younger embryos appears to be more convenient for women over 35 years.
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