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.

Artificial reproduction of the indoor-cultured brackish form of maraena whitefish (Coregonus maraena) under recirculated aquaculture system (RAS) conditions.

Maraena whitefish (Coregonus maraena) is a promising coldwater aquaculture species, especially in terms of meat quality. This versatile whitefish species, which inhabits both fresh and brackish waters, including the Baltic Sea, offers promising prospects for aquaculture breeding and cultivation. However, the main problem is the lack of availability of spawners of this species. Therefore, in this study, an attempt was made to raise the spawning stock under controlled conditions and then artificially reproduce the fish to obtain high-quality gametes and then larvae. The conducted research showed that successful culture of maraena whitefish spawners is possible under recirculating aquaculture system (RAS) conditions. The application of proper nutrition and manipulation of environmental conditions allowed the breeding of sexually mature whitefish spawners in less than two years in captivity (meat weight 476 g). Through stimulation of environmental conditions (temperature and photoperiod), it was possible to stimulate the final maturation of gametes of the F1 generation of cultured fish. The use of Ovaprim hormonal stimulation induced 100% of females to ovulate within 10 days of injection, compared to 60% of females in the control group. Additionally, the other examined parameters, such as the pseudogonado-somatic index (PGSI: 11.46% vs. 6.80%), fertilization rate (94.2% vs. 60.1%), embryo survival rate (92.6% vs. 41.1%) and embryo survival rate to the eyed-egg-stage (90.0% vs. 28.0), were significantly higher in the hormonally induced group, finally two times and three times, respectively. The mortality rate of spawners, after spawning and within three months of artificial spawning, was extremely low. The results obtained from the study demonstrate the feasibility of culturing the spawning stock of maraena whitefish in captivity and successfully inducing artificial spawning.

Effect of Different Water Salinities on the Larvae of the Blue Bream Ballerus ballerus (Linnaeus, 1758) during Rearing.

The influence of water salinities of 3, 5, and 7 ppt on the growth and survival of Ballerus ballerus (L.) larvae was studied. The control group was fish reared in freshwater (0 ppt). The larvae showed high tolerance to water salinities of 5-7 ppt. The mean final weight of the larvae ranged from 48.6 to 64.1 mg, with corresponding mean total lengths from 18.9 to 22.6 mm, depending on the water salinity level. The best larval length increments were recorded in water with salinity of 3 ppt. They were only slightly lower in 0 ppt water, and there were no statistically significant differences between the breeding rates calculated for larvae reared in 3 ppt water. Depending on the salinity level of the water, the final survival rate of the blue bream larvae ranged from 83.5 to 98.6%. The blue bream larvae reared in water with salinity levels of 5 and 7 ppt were statistically smaller than the others, but the results obtained were equally satisfactory.

Influence of Water Salinity on the Growth and Survivability of Asp Larvae Leuciscus aspius (Linnaeus, 1758) under Controlled Conditions.

The effect of water with a salinity 3, 5, 7, 9 and 11 ppt on the growth and survivability of asp Leuciscus aspius (L.) larvae was investigated. A control sample consisted of asp larvae reared up in freshwater (0 ppt). Larvae were fed for 21 days with nauplii of the brine shrimp Artemia salina. Water salinity was observed to have a considerable effect on the growth and survivability of asp larvae. In addition, saline water extended the life span of Artemia salina nauplii, which resulted in their prolonged availability to asp larvae. Asp larvae showed low tolerance to the salinity of water, reaching 9-11 ppt. Depending on the degree of salinity, the mean final weight of larvae varied from 122.6 to 139.4 mg, at body lengths from 23.8 to 25.6 mm, respectively. The best body length increments were recorded among asp larvae maintained in water with a salinity of 3 ppt. Depending on the level of water salinity, the final survivability of asp larvae ranged from 16.9 to 94.5%. The highest and increasing mortality was demonstrated among the larvae reared in water of the salinity equal to 11 ppt. It is not recommended to rear asp larvae in water with a salinity above 7 ppt due to the low survivability and large differences in the body size of the larvae that managed to survive.

Comparison of different methods of wild vimba bream Vimba vimba (L.) spawning under controlled conditions.

This paper describes a study in which different types of the reproduction of wild vimba bream (Vimba vimba) under controlled conditions were compared. Reproduction was performed by using three methods: spontaneous (natural spawning- NS groups): without hormones but with nests, semi-artificial (S groups) with hormones and nests, and artificial (A groups) with hormones and manual stripping of gametes. In groups S and A, different types of hormonal stimulation were tested: common carp pituitary homogenate (CPH), human chorionic gonadotropin (hCG), and commercial pellets containing mammalian GnRHa and dopamine antagonist-metoclopramide (Ovopel). The number of spawning fish, i.e., spermiating males and ovulating females, were compared depending on the type of breeding and the stimulant used. The lowest number of finally matured fish: spermiating males (67 %) and ovulating females (0 %) were noted in the spontaneously reproduced group. The best method of reproducing wild vimba bream under controlled conditions was the semi-artificial approach with hormonal stimulation and spawning nests, where ovulation rates in hormonally treated groups were between 43 % and 71 %, vs 26-57 % in artificial spawning groups. No ovulation observe in control groups in S and A types of reproduction. The highest embryo survival to the eyed-egg-stage was noted in group A-hCG (average 88.2). A higher value of spawning effectiveness (Se) with the use of the same hormones was obtained for groups S (0.374-0.574) than for groups A (0.252-0.493). In both cases, the highest Se was recorded for Ovopel stimulated fish and the lowest in hCG stimulated groups.

Rearing of Bitterling (Rhodeus amarus) Larvae and Fry under Controlled Conditions for the Restitution of Endangered Populations.

Among the several dozen European freshwater fish species, only European bitterling (Rhodeus amarus Bloch) and Rhodeus meridionalis belong to the group of ostrakophilous fish. The embryonic and larval development of the fish in this reproductive group until the time of the yolk sac resorption takes place in the gill cavity of river mussels (Anodonta sp. or Unio sp.). This paper presents the results of the European bitterling Rhodeus amarus being reared under controlled conditions. Bitterling larvae were caught together with river mussels in the natural environment and subsequently placed in a tank for behavioural observations. Bitterling larvae were seen swimming in the water within a week of placing the bivalves under controlled conditions. The bitterling larvae were 8.6 ± 0.11 mm long when they started to swim actively. The rearing was conducted in water at 20 and 26 ± 0.5 °C and lasted for 6.5 months (200 days) in both variants. Initially, the larvae were fed with live nauplii of Artemia salina and subsequently with fodder. The bitterlings in tanks with water at 26 ± 0.5 °C were 66.2 ± 3.0 mm long and weighed 3389 ± 548 mg. For comparison, bitterlings kept in water at 20 ± 0.5 °C were 64.48 ± 3.4 mm long and weighed 3242 ± 427 mg. No larval malformities or mortality were observed during the larvae and fry rearing. The bitterlings had well-developed secondary sexual characteristics and exhibited pre-spawning behaviour at the end of the rearing. This produced suitable bitterling stocking material to be used in the conservation of small or endangered populations.

Artificial Reproduction of Blue Bream (Ballerus ballerus L.) as a Conservative Method under Controlled Conditions.

The blue bream Ballerus ballerus (L.) is one of two species of the Ballerus genus occurring in Europe. The biotechnology for its reproduction under controlled conditions needs to be developed to conserve its local populations. Therefore, a range of experiments were performed to obtain valuable gametes from blue bream following hormonal stimulation with carp pituitary homogenate (CPH), Ovopel and Ovaprim. CPH and Ovopel were injected twice and Ovaprim-once, under a pectoral fin. The spawners were inspected 12 h after the last injection, and the gametes were collected. Eggs were collected from spawning females and fertilized with sperm from three males. The smallest number of ovulating blue bream (four females) was obtained from individuals stimulated with Ovaprim. There were six to nine ovulating females in the other groups, accounting for 60-90% of the females in the respective groups. The latency period was the shortest in the spawners stimulated with Ovopel. The fish in this group ovulated 14 h after the hormonal injection. Spawning was synchronized and all females spawned simultaneously. The highest average weight of eggs per fish (75.4 g) was obtained from the females stimulated with Ovopel, whereas the individuals stimulated with Ovaprim provided the smallest average amount of eggs (49.5 g). The study showed that blue bream spawners are not very sensitive to reproduction-related handling under controlled conditions.

Gonadal steroid fluctuations and reproduction results of European grayling (Thymallus thymallus L.) from broodstock farmed in a recirculation aquaculture system.

The initial experiment of this study was conducted to determine whether sex steroid concentrations varied before, during and after the reproductive season of European grayling (Thymallus thymallus L.) farmed broodstock confined in a recirculation aquaculture system (RAS). The results indicated that the plasma sex steroid (testosterone - T, progesterone - P4 and estradiol - E2) concentrations varied (P < 0.05) during these reproductive periods. There were greater concentrations of T, P4 and E2 before and during the grayling reproductive season indicating there are important functions of these steroid hormones associated with gonadal maturation, gamete production and reproductive behavior during the spawning period. In the second experiment of this study, the effectiveness of European grayling controlled reproduction treatment regimens was analyzed and there were 34.69 % and 68.18 % having ovulations in the 2 and 3-year-old broodfish, respectively. Of the embryos developing to the eyed-egg stage, there were 3.70 % and 6.87 % derived from eggs of 2- and 3-year-old grayling females, respectively. Proportions of embryos developing from the eyed-egg stage to hatching were 38.1 % and 52.1 % from eggs of 2- and 3-year-old grayling females, respectively, when there was culturing in a RAS. The results indicate that with grayling broodfish there is greater efficacy in induction of reproduction when there is imposing of the controlled reproduction treatment regimen on 3- rather than 2-year-old broodfish.