Darbo tikslas: įvertinti skirtingais racionais šertų veislinių bulių spermos kokybę. Darbo uždaviniai: 1. Išnagrinėti ir įvertinti skirtingus veislinių bulių racionus; 2. Nustatyti tiriamųjų bulių spermos kokybę; 3. Įvertinti skirtingų racionų įtaką veislinių bulių spermos kokybiniams rodikliams. Išvados: 1. B fermos buliai reproduktoriai gavo 16,4 MJ AE, 52,35 žaliųjų baltymų, 20,66 g fosforo, 40,9 mg vario, 859,2 mg cinko, 28290 TV vitamino D bei 122700 TV vitamino A daugiau nei A fermos buliai, kur šios medžiagos yra ypač svarbios reprodukcinei sistemai ir spermos kokybei. 2. A fermoje laikomų bulių išskirtas spermos tūris buvo 5,60 (±0,11) ml, koncentracija 1287,61 (±38,04) mln/cm3, gyvybingų spermatozoidų 81,22 (±0,57) proc., tiesiai judančių spermatozoidų 65,73 (±0,76) proc. B fermoje laikomų bulių vidutinis išskiriamas spermos tūris buvo 8,50 (±0,25) ml, koncentracija 1371,54 (±67,07) mln/cm3, gyvybingų spermatozoidų 80,13 (±0,37) proc, tiesiai judančių spermatozoidų 67,81 (±0,47) proc. 3. B fermoje laikomų bulių išskirtas spermos tūris buvo 2,90 ml (P<0,001), koncentracija – 84,92 mln/cm3 (P>0,05), tiesiai judančių spermatozoidų – 2,07 proc. didesnis (P>0,05) nei A fermoje laikomų bulių. Vienintelis priešingai rodęs rodiklis, t.y. gyvybingi spermatozoidai buvo 1,09 proc. (P<0,05) mažesnis už A fermos bulių. 4. Bulių davusių iki 4 ml spermos kiekį, koncentracija vidutiniškai nuo 1351,11(P<0,001) (A buliai) ir 2218,25 (P<0,001) (B buliai) sumažėjo iki 1097,43 (P<0,001)... [toliau žr. visą tekstą] / The aim of this research is evaluate the quality of bull semen, that are fed different diets. Research goal of the paper: 1. Analyze and evaluate different breeding bull rations; 2. Analyze bull’s semen quality; 3. Evaluate the influence of different diets on breeding bull semen quality (semen volume, concentration, percent viable sperm, directly moving spermatozoa percent). The results of different bull’s diet show that bulls of farm B got more mineral and vitamin needed for reproductive system than bulls from farm A. The breeding bulls of farm B were given 16.4 MJ 52.35 crude protein, 20.66 g of phosphorus, 40.9 mg of copper, 859.2 mg of zinc, 28,290 IU of vitamin D and 122 700 IU of vitamin A more than bulls from farm A. This material is particularly important for the reproductive system and sperm quality. Breeding bull’s sperm quality indicators showed that the semen characteristics (volume, concentration, and right-moving sperm) were on average superior to those bulls whose diets were additionally given mineral - vitamin supplements. The semen quality of bulls is changing dependent on different bull semen volume released. It showed that increasing volume of the sperm determine decreasing in sperm concentration mln/cm3.

Diss. (sammanfattning) Swedish University of Agricultural Sciences : Sveriges lantbruksuniv., 2006. / Härtill 4 uppsatser.

Universiẗat, Diss., 2003--München.

In the present study, we examined several cryoextenders previously used by several authors and various freezing protocols to determine the relative importance of each parameter on sperm freezing. The effects of controlled seeding and changes in cooling rate at different stages of freezing were also examined. Sperm samples from seven individual carp males were frozen in 0.5 ml straws by conventional freezing. Cooling rates were determined by monitoring the sample's internal temperature. We compared four freezing protocols, which involved placing sperm samples at various levels (1, 3, 6, and 9 cm) above the liquid nitrogen (LN) surface (corresponding to -190, -150, -110, and -70 °C, respectively) for 20 min followed by transferring the samples into LN. Freezing at 3 cm above the LN surface resulted in the highest motility (33 ? 8 %) and velocity (118 ? 9 ?m/s) of spermatozoa after thawing and diluting in swimming medium. We determined that -90 °C is an optimal temperature at which immersing the samples in LN does not affect sperm motility after thawing. The sperm motility of samples immersed in LN before or immediately after the crystallisation point (-16 °C) was 0 %. Motility of spermatozoa cryopreserved with or without a seeding procedure was not significantly different after thawing. Therefore, we hypothesise that supercooling the sample during the conventional freezing procedure is not the main damaging factor during carp spermatozoa cryopreservation.

The current study attempted to shed light on the regulatory processes and response arrangements of fish spermatozoa during the course of maturation and motility initiation. The first intent of this study was to improve the understanding of the mechanism underlying the acquisition of potential for sperm motility in sturgeon. Up to present work, the physiological process underlying sperm maturation in this species has not been described at all. Our results showed that sperm maturation in sturgeon occurs outside the testes because of dilution of sperm by urine and involves the participation of high molecular weight substances as well as calcium ions present in seminal fluid and/or urine. The second aim of the present study was to investigate the coping mechanisms in fish spermatozoa with osmotic and ionic activating mode, as well as in spermatozoa of euryhaline fishes, to various environmental conditions. We showed that alteration of environmental osmolality might affect the fish sperm in different ways, depending on fish species and modes of spermatozoa motility activation either osmotic or ionic mode. In response to osmotic stress caused by hypotonicity, carp spermatozoa regulated the flow of water across their cell membrane and increased their cytoplasmic volume during their short motility period. In contrast, no indications of sperm volume changes were observed neither in sterlet nor in brook trout spermatozoa, both of which having an ionic mode of motility activation. We also examined the mechanism by which sperm motility triggering in euryhaline fishes can adapt to a broad range of environmental salinity. Our results demonstrated that spermatozoa of euryhaline tilapia, Sarotherodon melanotheron heudelotii, reared in fresh-, sea- or hypersaline water can be activated in hypotonic, isotonic or hypertonic conditions of swimming milieu, provided Ca2+ ions are present at various levels. It was established that the higher the fish rearing salinity or the more hypertonic ambient media at spermatozoa activation, the higher extracellular concentration of Ca2+ ions is required. The results obtained in the present study allow suggesting that osmolality is not the main factor inhibiting sperm motility inside the testis in the S. melanotheron heudelotii. A third aim of this study was investigation of the regulation of motility initiation process and description of flagellar beating initiation in chondrostean spermatozoa. We detected that K+ inhibition of sperm motility in sturgeon can be by-passed due to the pre-exposure of sperm cells to a high osmolality shock prior to its transfer to K+-rich swimming media. Thus, we hypothesized that sturgeon spermatozoa may be activated by use of an unexpected signaling pathway, independent from regular ionic stimulation. The successive activation steps in sturgeon spermatozoa were investigated by high-speed video microscopy, using specific experimental situation, where sperm motility initiation was delayed in time up to several seconds. At motility initiation, the first couple of bends formed at the basal region begins to propagate towards the flagellar tip, but gradually fades when reaching the mid-flagellum. This behavior repeats several times until a stage where the amplitudes of bends gradually reach similar value, what eventually leads to sperm progressive displacement. The total period needed for the flagellum to switch from immobility with rigid shape to full activity with regular propagating bends ranges from 0.4 to 1.2 seconds. In conclusion, the results of the current study bring valuable pieces of information into the general understanding of the processes of maturation of fish spermatozoa, their adaptability to different physical and biochemical circumstances, the extra- and intra-cellular signaling as well as the regulatory mechanisms of motility activation in fish spermatozoa.

The whole Ph.D. thesis includes in total 10 chapters on 146 pages. Chapter 2 9 are specific parts of thesis where following scientific and practical aspects of pikeperch reproduction biology and aquaculture are described. Quality and quantity of pikeperch spermatozoa after varying cold water treatments are presented in the chapter 2 and can help to optimize broodstock management of males with the aim to obtain high quality spermatozoa during a seasonal and an out of season spawning as well. Benefits of hormone treatment of both sexes in semi-artificial reproduction in pikeperch are described in the chapter 3 where the importance of hormonal treatment of both sexes in tank spawning is evaluated for effective production pikeperch larvae. The use of an alcalase treatment for the elimination of pikeperch egg stickiness is being shown in the chapter 4 of this thesis. Post-ovulatory oocyte ageing and its effect on eggs viability rates and occurrence of larval malformations and ploidy anomalies are listed in chapter 5. These results describe the effects of the egg over ripening fertilization process in pikeperch. The first report of heat shock triploidisation in pikeperch is described in chapter 6 with production of 100% pikeperch triploid population. The effect of water surface treatment on survival, swim bladder inflation and growth of larvae is given in the chapter 7 with the aim to optimize the intensive culture of pikeperch larvae. Last two chapters (8 and 9) are describing the adaptation of intensively cultured juveniles to pond culture and the adaptation of pond-cultured juveniles to RAS as a new and effective methods for ongrowing production of pikeperch. In total, four published scientific papers, one handbook, one accepted scientific paper for publication, and two prepared scientific manuscripts are included and discussed in this Ph.D. thesis.

Pollution of the aquatic environment by inorganic and organic chemicals is a major factor posing a serious threat to the survival of aquatic organisms including fish. In addition balancing risks and benefits of fish consumption is nowadays an intensively discussed public health topic. Spermatozoa of almost all fish species are released into water environment where they can be directly exposed to various compounds, such as xenobiotics including toxic metals, prior to fertilization. In addition, exposure of parental adults to various xenobiotics may affect gamete quality, which may subsequently reduce fertilization success. On the other hand the advantages of eating fish are well-known, not only in the point that fish is a healthy source of protein and other nutrients, but eating contaminated fish may also confer various health benefits. Research over the past few decades has shown that the nutrients and particularly the n-3 fatty acids (FA) found in fish and seafood, are for examples protective against cardiac diseases and have a positive impact on brain development. The thesis provides a focus on two different cell model types. Firstly, human hepatocellular cells (Hep G2, ATCC) were used as in vitro tool for studying the effect of the intake of cadmium (Cd2+) contaminated fish on cytotoxicity, oxidative stress and fatty acid and phospholipid class compositions. Secondly, spermatozoa of one threatened species of fish, sterlet (Acipenser ruthenus) were used as in vitro model for studying effect of potentially hazardous xenobiotic compounds' occurring in open waters. Sperm from sterlet were exposed for 2h to environmentally relevant concentrations of DQ (0-150

<p>Ve&scaron;tačko osemenjavanje (VO) je najznačajnija reproduktivna biotehnologija u<br />intenzivnoj proizvodnji svinja. Efikasan izbor visoko fertilnih genetski superiornih<br />nerastova i visok fertilitet ve&scaron;tački osemenjenih krmača, ima veliki ekonomski uticaj na<br />efikasnost praktične primene ove biotehnologije. Međutim, prethodna istraživanja<br />pokazuju da procena klasičnih parametara fertiliteta ejakulata (koncentracija, ukupan<br />broj, pokretljivost i morfologija spermatozoida) nisu dovoljni pokazatelji fertiliteta i<br />reproduktivne performanse nerastova. Sa druge strane, pokazalo se da je fertilitet<br />ve&scaron;tački osemenjenih krmača, često, niži od onog kod prirodno osemenjenih krmača.<br />Kao osnovni razlog nižeg fertiliteta kod VO krmača, navodi se osemenjavanje sa<br />prekomerno razređenim dozama i/ili dozama dugotrajno čuvanim (3 do 5 dana).<br />Rezultati prethodnih istraživanja ukazuju da komponente semene plazme imaju ključni<br />uticaj na fertilizacioni potencijal spermatozoida in vivo i in vitro, kao i na fiziolo&scaron;ke<br />procese važne za uspe&scaron;nu oplodnju i razvoj embriona u uterusu.<br />Zbog toga je glavni cilj ovog istraživanja bio da se: (a) odredi sadržaj proteina u<br />spermalnoj plazma nerastova koji se koriste za VO na nekoliko komercijalnih farmi<br />svinja u Srbiji, (b) oceni uticaj sadržaja proteina u spermalnoj plazmi na pokretljivost i<br />morfologija spermatozoida u nativnoj i razređenoj spermi, nakon 3 dana čuvanja u<br />razređenom stanju i (c) ispita uticaj intrauterine infuzije spermalne plazme, pre<br />aplikacije klasične VO doze, na fertilitet krmača.<br />Sadržaj proteina u spermalnoj plazmi se kretao između 1% i 6,5%, &scaron;to je<br />ustanovljeno kod 212 uzoraka, dobijenih iz ejakulata 106 nerastova, koji se koriste za<br />VO na 6 farmi u AP Vojvodini. Nizak nivo proteina (1-3.5%, prosečno 2,4%) je<br />ustanovljen kod 69%, a visok nivo proteina (3.6-6.5%, prosečno 4,2%) kod 31%<br />ispitivanih nerastova. Nije ustanovljen značajan (P&gt;0,05) uticaj rase ili starosti nerasta,<br />kao ni godi&scaron;nje sezone, na sadržaj proteina u spermalnoj plazmi. Citomorfolo&scaron;ka<br />svojstva spermatozoida su testirana sistemom CASA i protočnom citometrijom. U<br />nativnoj spermi testiranih nerastova, prosečno je bilo 71% živih, 13% spermatozoida sa<br />o&scaron;tećenim akrozomom i 32% spermatozoida sa morfolo&scaron;kim anomalijama. Volumen<br />ejakulata, koncentracija, ukupan broj i pokretljivost spermatozoida bili su značajno<br />(P&lt;0,01) veći kod nerastova sa visokim, u poređenju sa niskim sadržajem proteina u<br />spermalnoj plazmi. Progresivna pokretljivost - PP (64%) i broj živih spermatozoida - ŽS<br />(66%) bio je značajno veći, dok su broj spermatozoida sa o&scaron;tećenom ćelijskom<br />membranom - OM (19%), akrozomom - OA (29%) i hromozomima - OH (13%) bili<br />značajno (p&lt;0,01) niži u uzorcima sperme sa visokim sadržajem proteina, koji su bili<br />čuvani 72h u razređenju 1:4, od ovih vrednosti kod uzoraka sa niskim sadržajem<br />proteina (PP = 48%, ŽS = 44%, OM = 27% OA = 45% i OH = 22%). Zamena autologne<br />spermalne plazme iz ejakulata sa niskim sadržajem proteina od jednog nerasta, sa<br />homologom spermalnom plazmom iz ejakulata drugog nerasta sa visokim sadržajem<br />proteina, značajno (p&lt;0,01) povećava progresivnu pokretljivost spermatozoida, sa 52%<br />na 65%, kod uzoraka čuvanih 72h u razređenju 1:4. Intrauterina infuzija 30 ml semene<br />plazme, pre aplikacije klasične VO doze, značajno (p&lt;0,05) povećava vrednost<br />pra&scaron;enja (94%) i prosečan broj živo rođene prasadi po leglu (12.3) (p&lt;0,01), u<br />poređenju sa kontrolom grupom krmača (83% i 10.5 prasadi).<br />Na osnovu dobijenih rezultata, može se zaključiti: (1) postoje značajne razlike u<br />sadržaju proteina u spermalnoj plazmi između pojedinih nerastova, (2) uzorci sperme sa<br />visokim sadržajem proteina, imaju veće vrednosti fertilizacionog potencijala<br />spermatozoida, od uzoraka sa niskim sadržajem proteina u spermalnoj plazmi, posle 72h<br />čuvanja u razređenju 1:4 i (3) infuzija spermalne plazme, pre aplikacije klasične VO<br />iv<br />doze, značajno povećava fertilitet tako tretiranih krmača, u poređenju sa kontrolnim<br />krmača. Ovi rezultati pokazuju da određivanje sadržaja proteina u spermalnoj plazmi,<br />može biti korisno sredstvo za predviđanje stepena fertiliteta nerasta, pre njegove<br />upotrebe za VO, kao i da se spermalna plazma može koristi za povećanje fertiliteta<br />ve&scaron;tački osemenjenih krmača. Dobijenim rezultatima su, u potpunosti, potvrđene radne<br />hipoteze i ostvareni postavljeni ciljevi istraživanja.</p>

Post-transcriptional regulatory mechanisms are widely used to influence cell fate decisions in germ cells, early embryos, and neurons. Many conserved cytoplasmic RNA regulatory proteins associate with each other and assemble on target mRNAs, forming ribonucleoprotein (RNP) complexes, to control the mRNAs translational output. How these RNA regulatory networks are orchestrated during development to regulate cell fate decisions remains elusive. We addressed this problem by focusing on Caenorhabditis elegans germline development, an exemplar of post-transcriptional control mechanisms. Here, we report the discovery of GLS-1, a new factor required for many aspects of germline development, including the oocyte cell fate in hermaphrodites and germline survival. We find that GLS-1 is a cytoplasmic protein that localizes in germ cells dynamically to germplasm (P) granules. Furthermore, its functions depend on its ability to form a protein complex with the RNA-binding Bicaudal-C ortholog GLD-3, a translational activator and P granule component important for similar germ cell fate decisions. Based on genetic epistasis experiments and in vitro competition experiments, we suggest that GLS-1 releases FBF/Pumilio from GLD-3 repression. This facilitates the sperm-to-oocyte switch, as liberated FBF represses the translation of mRNAs encoding spermatogenesis-promoting factors. Our proposed molecular mechanism is based on the GLS-1 protein acting as a molecular mimic of FBF/Pumilio. Furthermore, we suggest that a maternal GLS-1/GLD-3 complex in early embryos promotes the expression of mRNAs encoding germline survival factors. Our work identifies GLS-1 as a fundamental regulator of germline development. GLS-1 directs germ cell fate decisions by modulating the availability and activity of a single translational network component, GLD-3. Hence, the elucidation of the mechanisms underlying GLS-1 functions provides a new example of how conserved machinery can be developmentally manipulated to influence cell fate decisions and tissue development.

Diss. (sammanfattning) Uppsala : Sveriges lantbruksuniv. / Härtill 4 uppsatser.