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The Tench Fertility and Condition Changes in Fishery HLuboká
ŠAMPALÍK, Jan
The main present effort in the controlled fish reproduction is the finding universal preparation which are suitable for the ovulation induction of large fish species spectra. The preparation should not cause hight after-spawn mortality of generation fish and it is wantable to repeat inorganic spawn in following seasons too. The experiment which reviewed the tench fertility and condition with using several hormonal preparation was realized in working condition of Fishery Hluboká (hatchery Mydlovary) in 2006 {--} 2007. The gained generation material was separated in several intervals by weight and it was detected the working fertility in percent and semination in percent for individual weight categories. The experiment was realized by using following preparation: Ovopel, Dagin, Supergestran, Kobalerin.
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Vliv teploty na udržení schopnosti oplození a líhnivosti při přechovávání neoplozených jiker u lína obecného
ANDONIU, Andreas
This theses deals with the storage length of artificially spawn of hard roes of Tench (Tinca tinca) during different temperatures at the time before the semen discharging and activation to fertilization, hatching and consequent survival of fish hatchery throughout changeover from the embryonic to larval life period (beginning of active food intake). Homogeneous assortment of hard roes obtained from hormonally induced artificial hatching of 6 spawners has been used for this experiment. Samples of hard roes were put into plastic bowls and covered, immediately after artificial hatching. Subsequently, they were placed into tempered, thermo-isolating containers with temperatures of 5, 10, 15, 20, 25 and 30 °C. In time intervals of 0.5; 1; 1,5; 2; 3; 4; 6; 8 and 10 hours, a small amount of hard roes were taken away from each temperature (estimated 50 -100 pieces) and put into dry, glass beakers (in 3 repetitions in each temperature combination and length of storage). Subsequently, the semen discharging from 6 milters was carried out and activation by water was performed. Incubation took place in non-sticking environment. During the incubation, or more precisely during the consequent storage of embryos through temperatures between 19-20.5 °C, water was changed daily. Fertilization was evaluated 48 hours after fertilizing. Hatchery was determined 48 hours after beginning of hatching of first specimen. After changeover from embryonic to larval period of ontogenetic development, living food was offered to hatching fish (artemia sp.). Thereafter, the amount of hatched fish with filled intestines was counted. Ascertained values were depicted as a percentage from the total number of seeded hard roes as well as fertilized hard roes with the use of statistic methods (two factors Anovy with the repetition). The highest level (in statistic evaluation on the importance level alfa = 0.05) of hard roe hatchery was accomplished throughout the length of possession and temperature 1 hour/ 25 °C (68.0 +- 3.1 %). The high level of hatchery was maintained by hard roes stored for 2 hours, afterwards a gradual value decrease was registered. Similarly, that was achieved with hatching parameter, where the high level of hatching was achieved with hard roes possessed for the period of 3 hours (except temperature of 30 °C), afterwards the hatchery was decreased. Pursued survival and food intake parameters of hatched fish (from the practical point of view) confirmed above stated dispositions. The high hatchery from placed hard roes was maintained for 1.5 - 3 hours (except 30 °C), thereafter there was its gradual decrease. In the time of 8 hours (temperatures 5 - 20 °C), the survival of 1.2 +- 1.8 %, was found out, with the rest, the survival was nearly zero.
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Možnosti fixace vzorků pro měření obsahu DNA u ryb průtokovou cytometrií
HUBÁLEK, Martin
This thesis aims to assess the possibility of the usage of various biological fixatives for fish cell and tissues samples in order to extend its storage for later flow cytometric measurement of DNA content. The model species chosen were sterlet and tench, from which three types of samples were obtained: blood and fin tissue of subadult / adult individuals and tail tissue of hatched larvae. Altogether 13 fixation methods were tested for each type of sample of both model species. Methods were chosen based upon their easy feasibility and low time-consumption. The samples were measured on flow cytometer in native state immediately after sampling and placing in physiological saline and after 1, 5 and 10 days of fixation during which they were stored in a fridge or in a freezer at -80 ?C. Their analysis was carried out simultaneously with standards native cells from tench fin tissue when investigating sterlet samples, and commercially available fixed trout erythrocytes for tench samples. A fluorochrome used was 4',6-diamidine-2'-phenylindole dihydrochloride (DAPI; with excitation/emission maxima 358 / 461 nm). Based on the evaluation of coefficients of variation (CV) of fixed samples and the changes in their fluorescence levels in comparison with native state, optimal procedures for extended storage of all types of samples from both model species are suggested.
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Mikromanipulace a kryopreservace zárodečných buněk ryb
LINHARTOVÁ, Zuzana
The induction of germ-line chimerism is an expanding focus of fisheries research. This technique is having a potential to enhance the production of gametes of species that are commercially valuable, endangered, species with problematic reproduction, using a more common or easily available species or species adapted to artificial reproduction as a surrogate host. The main goal of this technology is to establish a small-bodied surrogate broodstock producing functional donor gametes based on germ cell transplantation. Extent preliminary experiments, including documentation of donor/host embryonic and larval development, characterization of germ cells enriched by documentation of their migratory activities, sterilization of the host, isolation and cryopreservation of donor germ cells, are key factors for launching this biotechnology. All these crucial points were the main objective of the present work. The whole thesis provided the focus on two different fish species. First, our commercially valuable fish, the tench, where we would like to apply our current knowledge to create a germ-line chimera within cyprinids by transplantation of tench germ cells to smaller and faster-reproducing fish species as white cloud mountain minnow. Secondly we focused on the endangered species (listed in IUCN Red List) of large body size with long reproductive cycle, the sturgeons. In this case, we have chosen sterlet as a host, providing an advantage of shorter generation interval and smaller body size, to produce gametes of donor, a critically endangered species of large body size with long reproductive cycle, such as beluga. This innovative technology could result in collection of sperm and eggs in shorter time from small-bodied host. In tench we firstly focused on embryonic and larval development documentation together with description of origin and migration pathways of primordial germ cells (PGCs). PGCs represent a powerful tool for creation a germ-line chimera within fish species because they transmit genetic information to the next generation (Linhartova et al., 2014a). Secondly, we reported a practical technique for isolation and cryopreservation of early stages of germ cells (GC), including spermatogonia (SG) and spermatocytes (Linhartova et al., 2014b). In case of sturgeons, Saito et al. (2014) firstly described the origin and migration patterns of sturgeon PGCs deposited at the vegetal pole of the egg similar to that in anurans. Secondly, Psenicka et al. (2015) reported isolation and cryopreservation of female and male GC, SG from testes and OG and pre-vitellogenic oocytes from ovary, of 2-4-year old Siberian sturgeon. Moreover the isolated GC were transplanted into host (sterlet) and process of transplantation resulted in successful colonization of sterlet genital ridge. The potential host for germ-cell tranplantation, sterlet, was sterilized by knock-down of germ cell specific gene, the dead end gene, by the morpholino antisense oligonucleotide (MO) agent (dnd-MO). These results reported the first known and functional method of sturgeon sterilization (Linhartova et al., manuscript). We provided important information on morphology and ultrastructure of beluga spermatozoa structure by scanning and transmission electron microscopy to increase knowledge of evolutionary and taxonomic relationships among sturgeons (Linhartova et al., 2013). Finally, this thesis presents several studies with differing focus of research but with one target goal to induce germ-line chimerism in fish. All these results are prerequisite of future application and development of surrogate production in these species.
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Impact of nutrition and rearing technology on the changes of the quality of common tench (Tinca tinca) meat
PŘÍBORSKÝ, Josef
The aim of the study was to determine the impact of diet (natural and formulated feed) on the chemical composition and fatty acids profile of the harvested fish. The content of dry matter in fish flesh resulting from the formulated diet was higher vs. the natural diet (23.94?1.24 % vs. 19.66?0.82 %) with nitrogenous compounds (60.24?2.82 % vs. 72.12?1.75 %), total fat content (24.81?4.51 % vs. 6.14?2.85 %) and ash (7.55?1.28 % vs. 10.54?1.53 %) respectively. The spectrum of fatty acids was determined by gas chromatography using Varian 3800 equipment. Tench fed on a formulated diet in the recirculating system had a significantly higher content (P< 0.05) of monounsaturated fatty acid (MUFA = 43.04?1.68 %) and n-6 polyunsaturated fatty acids (PUFA = 15.47?1.07 %) in their flesh compared to the flesh of fish reared in earth ponds on a natural diet - MUFA (32?5.29 %) and n-6 PUFA (13.6?1.66 %). Tench fed on a natural diet in earth ponds proved to have a significantly higher content (P< 0.05) of n-3 PUFA (16.8?4.38 %) and ? PUFA (30.3 ? 5.3 %) than tench reared in the recirculating system - PUFA n-3 (10.05?0.85 %) and ? PUFA (25.52?1.07%). The ratio n-3/n-6 for fish from earth ponds was 1.2; for fish from the recirculating system the ratio was 0.65. The results show a significantly higher composition of n-3 PUFA in flesh of tench from earth pond with natural food compared to fish on an intensive feeding diet in the recirculating system which showed a higher content of n-6 PUFA.
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