Effect of temperature and spawning agents on wild female asp (Leuciscus aspius) reproductive efficiency under controlled conditions.

Artificial reproduction is a bottleneck to produce stocking material for many species of freshwater fish. One of these species is the asp, Leuciscus aspius. Research in the field of artificial reproduction of this species is very scarce and often incomplete. There are no breeding protocols specifying optimal environmental conditions and hormonal stimulation for many species of rheophilic cyprinids, including asp. Since the number of natural asp populations is constantly decreasing, it is important to support natural stocks by restocking with high quality stocking material. For this reason, optimized protocols are needed to breed this species under controlled conditions to produce stocking material with high biodiversity and good health. Such an approach will make it possible to maintain the population of natural asp at a constant level. The aim of this study was to develop the protocol of asp artificial reproduction using optimized thermal conditions and appropriate hormonal stimulation. In experiment I, the influence of constant temperature (10.0, 12.0 and 14.0 °C) on the effectiveness of artificial reproduction of asp. In experiment II, the effectiveness of asp reproduction was checked after the application of spawning agents: Ovopel, Ovaprim or a combination of these two agents The obtained results indicate that for the final maturation of oocytes (FOM) and artificial reproduction of asp in controlled conditions, water temperatures of 10-12 °C are the most useful. Under these thermal conditions, the highest percentages of female's ovulation and embryo survival, as well as the percentage of hatching, were obtained. Hormone injections are necessary to perform final oocyte maturation (FOM) in female asp in captivity. All spawning agents used were especially useful for artificial reproduction of asp, however, the best values of the studied indices, such as ovulation rate and embryo survival, were obtained after the application of Ovaprim or the combination of Ovopel and Ovaprim in water temperature at a range of 10-12 °C. It was found that the pH of ovarian fluid may be a preliminary indicator of the biological quality of eggs in asps. The optimal pH value is 8.0-8.4. At pH below 7.4, no viable embryos were observed.

Growth Priorities of the Four Riverine Cyprinids during Early Ontogeny.

Growth patterns during early development of four species of riverine cyprinids, common dace (Leuciscus leuciscus L.), ide (Leuciscus idus L.), chub (Squalius cephalus L.) and common barbel (Barbus barbus L.), over 30 days of rearing, were analyzed. The study period covered three successive phases of larval development (preflexion, flexion, postflexion) and part of the juvenile development. The growth analysis carried out in the present study shows that significant allometric growth occurred mainly in preferentially localized body parts (head and tail) and was also associated with an increase in body height. The replacement of temporary structures (e.g., fin fold absorption) and the appearance of definitive structures around 21 mm TL (beginning of the juvenile period) corresponds to a stabilization of the relative growth of most parts of the body. This is probably due to the fact that the studied species had completed the most important part of the remodeling process (metamorphosis) and had achieved a body shape that allows juveniles to colonize faster water habitats. The results also indicate that individuals of species that are less developmentally advanced at the time of hatching (common barbel, chub) show greater initial dynamics of change in growth rate than individuals of species whose individuals are more advanced in ontogeny (common dace, ide). In addition, the small differences observed in growth parameters between successive periods analyzed, supported by the results of previous studies on these species, probably reflect the lack of abrupt changes in the behavior and habitat of the species studied.

Optimization of barbel (Barbus Barbus L.) fertilization and effects of ovarian fluid when there are controlled conditions for gamete activations.

Fertilization is one of the most important procedures in artificial reproduction and it directly affects the reproduction outcome. When there is optimization of fertilization, there can be a positive effect on subsequent reproductive processes and economic aspects of aquaculture. This study was conducted to determine time for which oocytes and sperm of barbel Barbus barbus retain fertilization capacity following placement in freshwater. Furthermore, the amount of ovarian fluid, excreted by fish during spawning with eggs (OFI; %) was determined, along with the chemical composition and effects on fertilization were determined. Gametes, ovarian fluid, and seminal plasma from barbel spawning specimens of the F4 generation were used to conduct the study. Ovarian fluid accounted for 14%-68% of contents of the mass released at spawning and post-spawning composition differed depending on whether hormonal treatments were utilized for control of reproduction. There was an association (R2 = 0.982; P = 0.000) between the pH of ovarian fluid and the barbel embryo survival rate. There was the greatest survival rate (>60 %) when the pH range of 7.9-8.4 and there was a lesser embryo viability when pH values were lesser or greater than values within this range (P <  0.05). The results from the study indicate that barbel eggs retain fertilization capacity longer (as long as 210 s) after activation by placement in fresh water than spermatozoa (about 30 s).

Inhibiting the influence of ovarian fluid on spermatozoa activation and spermatozoa kinetic characteristics in the common barbel Barbus barbus.

As a result of evolution, various finfish species have developed different breeding strategies. However, there are some similarities, and one of them is the positive effect of ovarian fluid on spermatozoa. The opposite of this phenomenon was found in the common barbel (Barbus barbus). The present study analyzed the effect of ovarian fluid (OF), distilled water (DW) and Woynarovich solution (WS) on the motility, longevity and kinetics of barbel spermatozoa. These spermatozoa parameters were also evaluated with various dilutions of ovarian fluid (OF) in relation to distilled water [0:4 (Group OF 0%), 1:3 (Group OF 25%), 1:1 (Group OF 50%), 3:1 (Group OF 75%), 4:0 (Group OF 100%)] and spermatozoa reactivation after a 30 s (Group OFR30s 100%) treatment in ovarian fluid. The motility analysis was carried out using computer-assisted semen analysis (CASA). The negative interaction of ovarian fluid with spermatozoa motility in the same fish species was recorded for the first time. In pure ovarian fluid, the average spermatozoa motility (MOT) decreased significantly (1.40 ± 0.94%). The negative effect of ovarian fluid-to-spermatozoa motility was reversible, and after a 30 s treatment in ovarian fluid and later dilution with water, spermatozoa motility was reactivated (from 2.25 ± 0.53% vs 69.78 ± 6.02%). The use of Woynarovich solution as an activator of spermatozoa movement had a positive effect (P < 0.05) on spermatozoa movement longevity (motility up to 90 s) and the percentage of motile spermatozoa compared to distilled water (up to 45 s) and ovarian fluid (P < 0.05).

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.

Early development and allometric growth in Nannacara anomala Regan, 1905 (Perciformes: Cichlidae) under laboratory conditions

Morphological development and allometric growth of laboratory reared Nannacara anomala were studied from hatching to the loss of larval characters and beginning of squamation (18 days post-hatching) at 26°C. The mean total length (TL) of larvae and juveniles increased from 3.74 mm at hatching to 9.60 mm at metamorphosis. Morphogenesis and differentiation were most intense during the first week of development. During this period (TL interval = 3.74 - 4.84 mm) there was an evident priority to enhance the feeding and swimming capabilities by promoting accelerated growth in the head and tail regions. Following this period, there was a major decrease in growth coefficients, indicating a change in growth priorities. Observations on the early development of Nannacara anomala confirmed the basic uniformity development of a substrate brooding cichlid...

Se realizó el estudio del desarrollo morfológico y crecimiento alométrico de Nannacara anomala criada en el laboratorio en la temperatura de 26°C, entre la eclosión y la pérdida de todas las características de larva y el principio del proceso de escamación (a los 18 días de la eclosión). La total logitud media (TL) de larvas aumentó de 3,74 mm en eclosión a 9,60 mm en el momento de metamorfosis. La morfogénesis y la diferenciación fueron más intensos durante la primera semana del desarrollo. En este periodo (el intervalo TL= 3,74 - 4,84) se observó una gran prioridad en la mejora de habilidades de encontrar alimento y nadar al promover el crecimiento de la zona de la cabeza y la cola. Transcurrido este periodo se notó una importante bajada de los coeficientes del crecimiento lo que indica el cambio de prioridades de crecimiento de distintas partes del cuerpo. Las observaciones del desarrollo temprano de Nannacara anomala confirman las similitudes principales en el desarrollo de cíclidos criados en sustratos...

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.

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%).