Effect of different treatment agents and post-treatment latency times on spermiation stimulation of northern pike (Esox lucius) under controlled conditions.

The effect of different treatment agents, namely, carp pituitary homogenate (CPH), Ovaprim ([D-Arg6, Pro9NEt]-sGnRH + domperidone) and a dopamine-receptor antagonist (metoclopramide), on the stimulation of northern pike (Esox lucius) spermiation was tested under controlled conditions. To carry out the experiment, males (n = 84) were divided into four groups: control (n = 21); CPH (n = 21); Ovaprim (n = 21); metoclopramide (n = 21). The control group was given 0.9% NaCl but no hormonal treatment. After 24 h, sperm was collected from seven males belonging to control (n = 7), CPH (n = 7), Ovaprim (n = 7) and metoclopramide (n = 7). This procedure was repeated after 48 and 72 h post-treatment. At each time, sperm was collected from seven males from each group only once. After collection, the quantity and quality of sperm were determined. It was confirmed that the treatment agent and latency time (the time between treatment and sperm collection) are two factors affecting the quantity and quality of northern pike sperm collected under controlled conditions. The highest total sperm volume and total sperm production (TSP) were noted in the CPH group compared to the Ovaprim, metoclopramide and control groups. In contrast, the time of sperm collection affected the sperm concentration (SC), TSP and sperm pH. With increasing time, SC and TSP decreased, which indicated the occurrence of sperm hydration being part of the final sperm maturation process. Sperm maturation is in turn a consequence of increases in sperm pH and seminal plasma osmotic pressure between 48 and 72 h post-treatment. Sperm motility and sperm kinetic parameters were affected by treatment agent and the time of sperm collection. This indicates that the sperm's ability to move that is achieved in the optimal environment (in spermatic ducts) is dependent on both factors which determine sperm maturation in northern pike under controlled condition.

Out-of-season artificial reproduction of common dace (Leuciscus leuciscus L.) under controlled conditions.

This study focused on the artificial reproduction of common dace Leuciscus leuciscus (L.) outside the reproductive season. The results indicate there is the possibility of initiating the reproducion in individuals of this before the natural spawning season. There could be induction of spermiation by altering the environmental conditions. For females, hormonal injections were necessary for induction of final oocyte maturation and ovulation. Generally, there were high percentages of spermiation (100%) and ovulation (87%-100%) as well as low mortality (0%-11% and 7%-13% for males and females, respectively) among the induced spawners. The greatest embryo survival occurred when Ovaprim (84.4%) or hCG (89.6%) was administered, although the latency time using hCG was at least twice as long compared to when other spawning agents were used (84-92 hrs and 30-44 hrs). The results from present study could be applicable for optimization of breeding stock numbers in aquaculture enterprises and in providing insights for conservation of L. leuciscus endangered populations.

Effect of carp pituitary homogenate (CPH) and sGnRHa (Ovaprim) on northern pike (Esox lucius) spermiation stimulation and its effect on quantity and quality of sperm.

The effect of carp pituitary homogenate (CPH, n = 7) at a dose of 2.0 mg kg-1 and (D-Arg6, Pro9NET)-sGnRH + domperidone (Ovaprim, n = 7) at a dose of 0.5 ml kg-1 in northern pike (Esox lucius) sperm maturation under controlled conditions was examined. On the control group, 0.9% NaCl at a dose of 1.0 ml kg-1 (n = 7) was used. Sperm was collected 48 h following injection. Sperm quantity (total sperm volume, total sperm production and sperm concentration), sperm motility using the Computer-assisted sperm analysis (CASA) system and sperm and seminal plasma quality (sperm pH, seminal plasma osmotic pressure and seminal plasma pH) were determined for each male separately in each group. The results of the present study demonstrate that hormonal treatment had a positive effect on sperm maturation in northern pike, regardless of the hormonal preparation used. However, even though no differences were found in total sperm volume and total sperm production between fish injected with either CPH or Ovaprim, it should be highlighted that the highest progressive motile sperm (PRG), straight-linear velocity (VSL) and movement linearity (LIN) was noted in fish treated with Ovaprim. It was also found that it is possible to collect sperm from non-hormonally manipulated fish. However, in such a case, only a small sperm volume (around 0.1 ml) characterised by lowered PRG (below 40%) was noted. Considering the fact that only after Ovaprim application sperm motility with progressive movement of sperm above 50% was observed, the hatchery practice of collection of sperm 48 h after its application (at a dose of 0.5 ml kg-1) may be recommended.

Enrichment of Zebrafish Danio rerio (Hamilton, 1822) Diet with Polyunsaturated Fatty Acids Improves Fecundity and Larvae Quality.

The zebrafish is a freshwater fish of the Cyprinidae family, which is frequently used in scientific research. It owes its popularity to its genome, whose structure is comparable to the human genome and, for this reason, this species is often used in human medical research. However, such research requires high-quality material to conduct tests producing repeatable results. This study examines the effect of providing feed enriched with essential fatty acids, especially docosahexaenoic acid, to zebrafish spawners on fertility as well as the survival rate and growth of their offspring. The experiments revealed a significant (p < 0.05) effect of feeding spawners with feed enriched with polyunsaturated fatty acids on all tested parameters: their fertility (30% higher compared to the control group), survival rate, and the posthatching size of larvae, which were larger by 13% and 4%, respectively, compared to the control group.

New technique for fertilizing eggs of burbot, asp and ide under hatchery conditions.

The development of a new protocol for egg fertilization may increase embryo survival and benefit the aquaculture process. In the present study, a new technique of partially adding sperm to activated eggs in the artificial fertilization of burbot (Lota lota), ide (Leuciscus idus) and asp (Aspius aspius) eggs was evaluated. If the same volume of sperm was divided into two or three parts and added to eggs in 30-60s intervals, it significantly improved embryo survival at the eyed-egg-stage of development. In the present study, the periodic addition of spermatozoa to eggs affected fertilization (ide and asp) and embryo survival rates (ide, asp and burbot) and might be successfully applied under hatchery conditions.

Use of an ultrasound device to determine sex and to perform gonad biopsy in the European eel Anguilla Anguilla / Uso de um dispositivo ultrasonográfico para determinar o sexo e efetuar a biópsia das gônadas da Enguia-Européia Anguilla anguilla

The European eel Anguilla anguilla is a highly important market species which is also increasingly in danger of extinction in Europe. One of the ways of protecting the species in the natural environment, while maintaining its market supply, is its aquaculture, e.g. due to much higher survival rate under controlled conditions than in the wild. However, this can be done only when an effective artificial reproduction biotechnique is developed. The aim of this study was to use an ultrasound device to determine the sex and to perform a biopsy of the gonads, which is a part of complete protocol of eel artificial reproduction. The findings indicate that ultrasonography is highly useful in determining the sex in migrating eel (100% of sureness) and in performing high-precision biopsy of the gonads. The present method gives the possibility to quick determine the maturity of the female gonads. The application of ultrasonography (USG) is totally safe for fish and a portable ultrasound device can be used in both the laboratory and in the field...

A Enguia-Européia Anguilla anguilla é uma espécie de grande importância comercial e que apresenta também grande risco de extinção na Europa. Uma das formas para a proteção das espécies em seu ambiente natural e de manter o seu suprimento no comércio é a aquicultura, por exemplo, devido à taxa de sobrevivência em condições controladas ser muito superior a existente em condições naturais. Contudo isto só pode ser obtido quando é desenvolvida uma biotécnica de reprodução artificial eficiente. O presente trabalho foi delineado para utilizar um dispositivo ultrassonográfico a fim de determinar o sexo e de possibilitar a realização de biópsias das gônadas, o que é parte do protocolo complete de reprodução artificial de enguias. As constatações indicaram que a ultrassonografia é muito útil para a determinação do sexo das enguias (100% de exatidão) e também é capaz de possibilitar a realização de uma biópsia das gônadas de alta precisão. O método descrito oferece possibilidades para a rápida determinação da maturidade das gônadas das fêmeas. A aplicação da ultrassonografia é totalmente segura para peixes e o dispositivo portátil de ultrassonografia pode ser utilizado tanto no laboratório como em condições de campo...

Effect of temperature on the effectiveness of artificial reproduction of dace [Cyprinidae (Leuciscus leuciscus (L.))] under laboratory and field conditions.

This study sought to determine the effect of water temperature on the effectiveness of artificial reproduction of dace brooders under laboratory and field conditions. Three temperatures were tested in the laboratory: 9.5, 12 and 14.5 °C (± 0.1 °C). The water temperature under field conditions was 11.0 ± 0.3 °C (Czarci Jar Fish Farm) and 13.2 ± 1.4 °C (Janowo Fish Farm). The study showed that artificial reproduction of dace is possible in all the temperature ranges under study and an embryo survival rate of over 87% can be achieved. Dace has also been found to be very sensitive to rapid temperature changes, even within the temperature ranges optimal for the species. Such changes have an adverse effect on the outcome of the reproduction process, such as a decrease in the percentage of reproducing females, a decrease in the pseudo-gonado-somatic index (PGSI) and a decrease in the embryo survival rate.

Ovarian alterations in wild northern pike Esox lucius females.

The aim of the present study was to analyse the occurrence of macroscopically visible ovary alterations in 2 populations of northern pike Esox lucius L. originating from lakes in the Mazurian Lake District (NE Poland). The alterations were characterised by ovary tissue that was morphologically malformed, in part or in whole, and contained immature oocytes, i.e. trophoplastic or previtellogenic oocytes instead of vitellogenic oocytes. These alterations were found only in the ovaries, and no morphological alterations of the testes were noted. Macroscopic and histological analyses were carried out in order to classify the observed alterations in the ovaries. Three types of alterations were identified in which morphological malformations as well as histological investigation of the ovaries were considered. An analysis of the size and age of the fish in relation to the occurrence of alterations as well as of the macroscopic and histological nature of the alteration types was made. The data obtained revealed no lake or age dependency of the observed alterations. Based on the results obtained, we suggest that the presence of endocrine disruptors in the environment or/and genetic factors could be responsible for these kinds of gonad anomalies. However, our results did not allow us to determine the aetiology of the alterations.

Cortical reaction as an egg quality indicator in artificial reproduction of pikeperch, Sander lucioperca.

The aim of this study was to investigate the process of the cortical reaction in eggs of pikeperch, Sander lucioperca (L.), as well as the application of microscopic assessment of this process in egg quality evaluation. The analysis was carried out with eggs obtained from 10 females by artificial reproduction, in which hormonal stimulation with hCG was applied. Subsequently, each sample of eggs (separately from each female fish) was analysed. The analysis included observation of the cortical reaction and the process of egg swelling, and determination of the effect of temperature (12, 14 and 16°C) and the presence of spermatozoa on the cortical reaction. The results indicate that the cortical reaction in pikeperch eggs is quite violent, resulting in visible deformation of eggs between 3 and 5 min after activation. No effect of temperature or the presence of spermatozoa on the cortical reaction was observed. A strong correlation was recorded for the percentage of egg deformations observed and embryo survival rate. The described method of determination of pikeperch egg quality (based on egg deformation rate between 3 and 5 min after activation) may be highly useful, both in scientific research (where high-quality eggs are required) and in hatchery practice.

A new classification of a preovulatory oocyte maturation stage suitable for the synchronization of ovulation in controlled reproduction of Eurasian perch Perca fluviatilis L.

To improve controlled reproduction of Eurasian perch Perca fluviatilis, the criteria for the evaluation of final oocyte maturation stages were revised. The new classification covers six preovulatory maturational stages (I -VI) from the end of vitellogenesis to germinal vesicle breakdown (GVBD) and was based on macroscopic changes of preovulatory oocytes (position of the germinal vesicle, GVBD, oil droplets coalescence). The observation was performed during out-of-season artificial reproduction with the use of a single hCG injection (500 IU/kg). The classification was subsequently verified with the controlled reproduction of wild female perch with the use of hormonal stimulation (500 IU hCG/kg of body weight at 12°C). The females were at different maturational stages and constituted respective experimental groups (I-VI). During the experiment, ovulation appeared to be considerably synchronized within particular groups. Statistical differences in latency time (time between hormonal treatment and ovulation) were found between experimental groups (mean latency time: 110, 92, 68, 49, 29 and 18 h in groups representing VI, V, IV, III, II and I stage of the proposed classification, respectively). The proposed classification and the results presented in the study allowed for effective synchronisation of ovulation. The use of our new oocyte maturation classification may positively influence the effectiveness of Eurasian perch production.

Artificial reproduction of wild and cultured barbel (Barbus barbus, Cyprinidae) under controlled conditions.

The aim of this work was to compare the effects of controlled reproduction of cultured and wild common barbel, Barbus barbus (L.). Preparations containing different GnRH analogues and dopamine receptor antagonists (Ovopel, Ovaprim) as well as human chorionic gonadotropin (hCG) (in the case of cultured fish) were applied and their influence on ovulation, spermiation and quality of gametes obtained was determined. No differences in the qualitative or quantitative parameters of semen were found between fish stimulated with different hormonal preparations and those not receiving hormonal stimulation. The high suitability of Ovaprim for ovulation induction in (cultured and wild) barbel was confirmed. The highest synchronisation of ovulation was obtained after the application of Ovopel (18 ± 3 h), but the best results of controlled reproduction (expressed as the percentage of ovulations and survival of embryos) were obtained by applying Ovaprim (83.2 ± 4.1). A significantly higher percentage of ovulation was obtained in cultured fish (80-90%) than in wild fish (< 25%).

Artificial reproduction of two different spawn-forms of the chub.

The aim of this study was to compare, under controlled conditions, reproduction results of cultured and wild stock of the chub. Wild fish spawned only once a season whereas the cultured stock spawned at least two times. In the multiple-spawn stock, fewer fish spawned and the weight of produced oocytes was reduced compared to the single-spawn stock. Larvae obtained from the multi-spawn forms were smaller than those of the single-spawn stock. The occurrence of one species with two forms of spawning performance in the same area makes it difficult to develop an efficient method for controlling the reproduction.

Reproductive parameters of common carp (Cyprinus carpio L) spawners during natural season and out-of-season spawning.

Common carp spawning characteristics were compared between season and out-of-season periods. There were no differences in the percentage of ovulation, spermatozoa motility and embryo survival to the eyed-egg-stage between the spawning periods. Fish spawned during the season produced oocytes of higher total weight than those spawned out-of-season. On the day exogenous feeding began, larvae obtained in out-of-season spawning were shorter than those obtained during the spawning season. However, the out-of-season larvae started feeding two days earlier than in-season larvae. As a result, seven days after fertilization, larvae obtained during out-of-season had the same length as larvae obtained during season. It appears to be possible to obtain common carp larvae half a year before the natural spawning period without decreasing the quality of gametes and larvae.