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Hodnocení kvality ejakulátu ovčáckých plemen psů
Leciánová, Tereza
The bachelor thesis deals primarily classified shepherd dog breeds in FCI classification and becoming familiar with it, which breeds belong there. The main part of this paper is to describe the anatomical and physiological characteristics of the reproductive organs of the dog - male sperm and a description of their development, ejaculation and semen collection. In conclusion mentions the progress of macroscopic and microscopic examination of fresh semen. The thesis will be followed by master's thesis, which will include the results and conclusions obtained during the examination.
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Frekvence výskytu morfologických změn spermií hřebců v Zemském hřebčinci Tlumačov s.p.
Jakubčíková, Simona
Morphological evaluation of sperm has a long history and is an essential element analysis of semen, which provides breeders invaluable information to assess the health status of animal and its potential fertility. An important role is played by the percentage of healthy sperm due to abnormally developed. This value plays an important role in the usability of quality in breeding stallions. Stallion sperm intended for artificial insemination breeding mares in native form may not contain more than 40 % of defective sperm and the total amount of their activity should not be below 60 %. In the case of benefits intended for deep freezing is allowed only 20 % pathological forms of sperm. Activity selected for cryopreservation of semen should not fall below 80 %. The topic of the thesis was to determine the frequency of morphologic sperm defects in a group of stallions stud farm Tlumačov s.p., included in breeding season for 2013. Sperm morphology examination was performed on 72 samples of ejaculates collected from 12 stallions during the breeding season from May to August. For each stallion sperm activity was recorded of subjective method, immediately after collection, and the correlation was observed with respect to morphological abnormalities of the same sample. To investigate the morphological structure of sperm abnormalities were stained with two methods of cytological stain -- staining of B.T.Farelly and staining of Čeřovský. After staining the preparations were evaluated under a light microscope. Identified defects were divided into 6 categories - defects in the head, acrosome defects, defects of the neck, tail defects, abnormal forms of sperm and immature sperm. We studied a number of normospermia and total sperm pathology also expressed as a percentage. Another part of the analysis was a theoretical comparison of both the staining techniques with respect to suitability for the examination of morphological sperm abnormalities in stallions.
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Biologie rozmnožování u raků
NIKSIRAT HASHJIN, Hamid
The ultrastructure of spermatozoa from six crayfish species were described. The acrosome complex and nucleus are located at the anterior and posterior of the spermatozoon, respectively. The acrosome complex organelle is divided into two main parts: the main body of the acrosome that is a dense inverted cup-shaped structure and organized into three layers of differing electron densities and extended parallel filaments, and the sub-acrosome zone occupying the central part of the acrosome complex, which is divided into two electron dense areas. The spermatozoon of Orconectes limosus is described for the first time. In addition, an acrosome spike in the spermatozoon of Procambarus clarkii is described. Despite a well conserved general structure and similarity of pattern among these spermatozoa, differences in the dimensions of the acrosome within the studied species may be useful to help distinguish the different crayfish. The ultrastructure of the spermatozoon and spermatophore wall of the narrow clawed crayfish Astacus leptodactylus during three stages including freshly ejaculated, post-mating, and after release of the spermatozoon were described and compared. The crayfish spermatophore consists of a sperm mass enveloped by a three layered spermatophore wall. After mating, the thickness of the outer layer of the spermatophore is increased. The matrix in the middle layer of the spermatophore becomes reticulated, and granules inside this layer release their contents. Fibers in the inner layer degrade to small particles. The spermatozoon capsule swells and space between the capsule and the spermatozoon appears. The area of the plasma membrane is increased by wrinkling of the surface and alteration from a single to a multilayered structure at the anterior part of the acrosome. The density of the subacrosome zone increases in the vicinity of the main body of the acrosome. With the onset of fertilization, the layers of the spermatophore are dissolved by female glair glands secretions. The capsule, plasma membrane, and membranous lamellae are eliminated, and bundles of filaments are released. The subacrosome zone loses electron density and retracts. The electron-dense material of the innermost layer of the acrosome is discharged and, together with filaments, forms a filament/droplet structure at the anterior part of the spermatozoon. The most important change is observed in the subacrosome zone, which may play a key role in the fertilization. Also, morphological changes of the spermatozoon after release from the capsule, especially formation of the filament/droplet structure, may contribute to the mechanism of egg-spermatozoon binding in the crayfish.
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Fish sperm motility parameters and total proteins profiles in seminal plasma during in vivo and in vitro storage
KOLEŠOVÁ, Anna
The effect of gamete storage on sperm quality has received considerable attention in recent years. Previous studies have shown that spermatozoa stored in vivo or in vitro for a long time can lost their motility and fertilization capacity. Moreover, it have been concluded that male fertilization potential is highly dependent, not only on spermatozoa motility parameters, but also on organic components including protein composition of seminal plasma. On the other hand, spermatozoa motility and protein profiles of seminal plasma are highly dependent on conditions of storage. Therefore, additional data about effects of in vivo and in vitro storage on quality sperm parameters and protein composition of seminal plasma are essential for development of fish artificial reproduction methods. In the current work the influence of in vitro and in vivo storage on parameters of sperm motility, DNA integrity, antioxidant defensive system and seminal plasma protein composition were studied. As a conclusion, the results of this study provide new data on sperm quality and quantity parameters of chondrostean and teleost fish species with respect to in vivo and in vitro storage capacities, which should be beneficial for the development of aquaculture of these species. The data confirmed that protein patterns in seminal plasma varied during in vivo storage, depending on time of sperm collection. Furthermore, the altered proteins are probably involved in enzymatic pathways that regulate spermatozoa movement. In practice, the results presented in this thesis should help to improve management and optimize the development of protocols for artificial reproduction.
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Optimalizace reprodukce a kvality gamet u okounovitých ryb
KŘIŠŤAN, Jiří
Pond and intensive aquaculture of percids relies mainly on juveniles supplied by semi-artificial and artificial reproduction. Fish hatcheries do not always provide optimal conditions for percid reproduction. This project was undertaken to optimize reproduction methods of Eurasian perch (Perca fluviatilis L.) and pikeperch (Sander lucioperca L.). Reproduction parameters were compared in farmed and pond-reared perch broodstock without hormone treatment. Spawning was successful without hormone induction at 92.3% and 76.9% in farmed and pond-reared perch, respectively. Significant differences in morphological characteristics (gonadosomatic, hepatosomatic and viscerosomatic indices), fecundity, hatching, and post-spawning mortality were detected. All results, with the exception of spawning success and post-spawning mortality, were superior in pond reared broodstock. Further studies are required to optimize protocols for using better artificial food in farmed perch and decreased stress and post-spawning mortalities in pond reared perch broodstock. The possibilities of using clove oil, Propiscin, or 2-phenoxyethanol as an alternative to tricaine methane sulphonate (MS222) as anaesthetic, particularly with respect to reducing fish stress, were investigated. The haematological and biochemical profiles of pikeperch anaesthetized with clove oil (33 mg l?1), Propiscin (1.5 ml l?1), 2-phenoxyethanol (0.3 ml l?1), and MS222 (150 mg l?1) were determined. Each tested group was divided into two subgroups; the first subgroup was sampled 10 min after application and the second 24 h post-application. Clove oil was shown to be associated with the lowest effects in pikeperch, and therefore could be recommended as an alternative to MS222, while Propiscin and 2-phenoxyethanol were shown to be unsuitable for use with pikeperch. In Eurasian perch the use of semi-artificial reproduction for mass production of embryos was shown to be simpler and more effective. With mass incubation, 1 134 000 embryos (59% of total production) were obtained with a semi-artificial spawning regime and 796 000 embryos (41% of total production) with artificial spawning. Based on these results, we can recommend the semi-artificial spawning for production of perch embryos. Differing doses of two commonly used hormone preparations: human chorionic gonadotropin (hCG) and mammalian GnRHa (Supergestran) were tested for induction of ovulation in pikeperch. A broad range of effective doses is used in aquaculture of pikeperch. The minimal effective dose based on ovulation success is still missing. Hormone therapy applied to excess can result a detrimental impacts on broodstock fecundity and quality of eggs. Results indicated that a single injection of hCG or Supergestran could be used for successful ovulation, while no spawning was observed in a control group treated by saline solution. Among hormone treated groups, the best results were observed in groups treated with hCG at 500 and 750 IU kg-1 BW and in groups treated with mGnRHa at 25 ?g kg-1 BW. The sperm of pikeperch and Eurasian perch show similar morphology and fine structure, with the exception of differences in arrangement of midpiece and centrioles and the presence of a fin-like structure on the flagellum in pikeperch. Our results suggest use of an activation medium containing Ca2+ for activation of short-term stored sperm. Efficacy of five concentrations of Alcalase enzyme for elimination of egg stickiness compared to a milk/talc suspension was determined. Results indicated that 0.5-2.0 ml l-1 of Alcalase for 2 min may increase pikeperch egg hatching success (80-85%) compared to the milk-talc suspension. The highest hatching rate (85.4%) was observed with 1.5 ml l-1 of Alcalase. The application of Alcalase successfully eliminated pikeperch egg stickiness in 2 min compared to 60 min with traditional milk/clay/talc methods.
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Efekt xenobiotik na DNA integritu a fyziologii rybích spermií
LINHARTOVÁ, Pavla
Pollution of the aquatic environment with xenobiotics has become a serious health concern in recent years. In the present study the effect of DQ, TBBPA, BPA and VIN on sperm quality parameters, DNA integrity and oxidative stress indices in sterlet (Acipenser ruthenus) sperm and sperm from brook trout (Salvenilus fontinalis) were investigated. To do this, an in vitro spermatozoa motility assay was used by a computer-aided Motion-Analysis system. The sperm of sterlet (Acipenser ruthenus) was diluted to obtain the spermatozoa density of 5×108 cells×ml?1 and then exposed for 2 h to final concentrations of xenobiotics: DQ - 25, 50, 100 and 150 ?M, TBBPA - 0.5, 1.75, 2.5, 5, 7.5 and 10 ?g/l, BPA - 0.5, 1.75, 2.5, 5 and 10 ?g/l and Vin - 0.5, 1.75, 2.5, 5 and 10 ?g/l. Spermatozoa velocity and percentage of motile sperm were significantly decreased at each time post-activation compare to control. The level of DNA damage expressed as a % DNA in Tail and Olive Tail moment significantly increased when spermatozoa were exposed to higher concentrations of xenobiotics. The level of oxidative stress indices lipid peroxidation (LPO) and carbonyl derivatives of proteins (CP) and antioxidant activity of superoxide dismutase (SOD) increased significantly with increasing concentration of xenobiotics. On the other hand the intracellular ATP content in sperm samples had a significantly decreasing effect. In short, xenobiotics can induce reactive oxygen species (ROS) stress in fish spermatozoa, which could impair the sperm DNA integrity, quality and antioxidant defense system. The present study confirms that environmental concentrations of xenobiotics are capable to induce oxidative stress, leading to impaired sperm quality, DNA fragmentation and intracellular ATP content Obtained results also suggested that the use of fish spermatozoa in vitro assays may provide a novel and efficiently means for monitoring residual pharmaceutical in aquatic environment.
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The electrostimulation the spermiation in the crayfish
KUBEC, Jan
The crayfish are important and indispensable animals in aquaculture. In the aquatic environment, they have a role as predators, food for other fish, detrivors and they are an important element in the circulation of the energy and the nutrient in the aquatic ecosystem. Today, number of the european crayfish species populations is decreasing. It is caused by unfavourable regulation of streams, water pollution and the occurence of crayfish plague. The main purpose of present study was examining the impact of electrostimulation on the males of the crayfish and obtaining sex products (the sperm). The principle of this research was determining whether the male of crayfish are able to extrude spermatophore with sperm by an electric stimulus. The transformer was used as resource alternating current in experiments. All experiments conducted in a laboratory of experimental device for fish farming in Vodňany. Experiments were conducted in 2011 on a male of the spiny-cheek crayfish (Orconectes limosus), the signal crayfish (Pacifastacus leniusculus) and the red swamp crayfish (Procambarus clarkii). The results of this work and success of the experiments suggest, that electrostimulation could be used for obtaining sex products (the sperm) from crayfish males. Results from a laboratory of electron microscopy confirm the harmlessness of electrostimulation on the shape and structure of sperm. The sperm can be used to artifical insemination of the crayfish eggs in future. However, sperm capacitation is problematic.
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