Electronic Journal of Polish Agricultural Universities (EJPAU) founded by all Polish Agriculture Universities presents original papers and review articles relevant to all aspects of agricultural sciences. It is target for persons working both in science and industry,regulatory agencies or teaching in agricultural sector. Covered by IFIS Publishing (Food Science and Technology Abstracts), ELSEVIER Science - Food Science and Technology Program, CAS USA (Chemical Abstracts), CABI Publishing UK and ALPSP (Association of Learned and Professional Society Publisher - full membership). Presented in the Master List of Thomson ISI.
Volume 7
Issue 2
Targońska-Dietrich K. , Zielazny T. , Kucharczyk D. , Mamcarz A. , Kujawa R. 2004. OUT-OF-SEASON SPAWNING OF CULTURED IDE (LEUCISCUS IDUS L.) UNDER CONTROLLED CONDITIONS, EJPAU 7(2), #02.
Available Online: http://www.ejpau.media.pl/volume7/issue2/fisheries/art-02.html


Katarzyna Targońska-Dietrich, Tomasz Zielazny, Dariusz Kucharczyk, Andrzej Mamcarz, Roman Kujawa



The comparison of out-off-season spawning of two cultured ide stock were examined. The obtained results show, that is possible to reproduced ide a few months before their natural spawning season. Generally, the percentage of spermiation and ovulation was very high, as well as spawners survival. The biological quality of the gametes was higher in stock cultured at ponds, than from fish cultured under controlled conditions. One ide females from cultured stock under controlled conditions ovulated without hormonal stimulation (control group). The quality of oocytes collected from this fish was much lower than from hormonally stimulated fish.

Key words: artificial spawning, Leuciscus idus, reproduction, out-off-season spawning..


The development of the civilization has contributed to modification of the water environment by building-up rivers as well as bringing significant amounts of contaminants into them. This state of matter has caused some serious inconvenience for existence and reproduction of the fish. Research conducted recently in polish rivers show changes in the fish species fauna. It applies especially to rheophylic fish [20, 25]. Ide (Leuciscus idus) is one of the species whose number has been subjected to systematic limitation for many years [1].

Experiments on reproduction of this species were made during the spawning season [2, 4, 5, 12]; there have also been some reports on obtaining gametes off the spawning season [11, 12, 13]. These experiments, however, were made on fish captured in natural environment. One of the biggest obstacles in artificial spawning of rheophylic cyprinid fish is getting the spawners [15]. There is very difficult to catch both sexes at the same area shortly before the spawning. Males reach the spawning area before females thus it would be more appropriate to catch the fish at the spawning areas or earlier, on the feeding territories [11]. These kinds of problems are solved by carrying out an artificial spawning with fish stocked in controlled conditions or in fish cultured in ponds [6, 26].

So far, however, there is a lack of such research related to ide. Drawing up a biotechnology of artificial reproduction basing on cultured fish will contribute to getting larger amounts of restock material. Currently there is often a shortage in it [1]. It results, among others, from getting insufficient amount of spawners from natural environment during the spawning season, or from insufficient amount of specimen of either sex.

The full study on methods of reproduction of the ide, regardless of the natural reproductive cycle, will enable getting more, and at the same time better prepared restock material for spring, which will increase the efficiency of restocking and will also allow breeding this species in aquaculture conditions no mater the season of the year. The aim of present study was out-off-season spawning of cultured ide.


Broodstock collection

One group of spawners was obtained from Krzelów Hatchery (south Poland), and then transferred to hatchery in Olsztyn. Second group of ide spawners was cultured in controlled conditions in the Department of Lake and River Fishery in Olsztyn. All fish were marked individually with plastic tags placed under their dorsal fin. Both, male and female spawners were being kept together in 1000-L tanks with controlled temperature and photoperiod [17].

Manipulations of environmental conditions

The experiment was conducted twice within the space of two years. The manipulations with environmental conditions were started at the beginning of October, what allowed to conduct the ide reproduction in January.

At the beginning, the temperature of water in tanks with spawners was set to 10°C, and the photoperiod (light: dark) to 10:14. During 6 week period the temperature was being reduced to the level of 2°C, and the time of lighting the reservoirs to 4 hours a day. The experiment proceeded with gradual increasing of the temperature of water in tanks (1, 5°C per week) up to 12°C. The tanks' lighting time was simultaneously extended to 12 hours a day.

Checking the oocytes maturation

Before approaching the propagation, maturation of gametes at spawners was checked each time. Oocytes were taken from females using the method described by Kujawa and Kucharczyk [16]. The oocytes samples were collected and placed in Serra’s solution for five minutes. Following the clarification of the cytoplasm the position of oocytes nucleus was defined according to four-degree scale:

Next stage of the experiment involved only females whose oocytes maturation was between 2-3 and 3 divisions. This is the best moment to collect cyprinids female for hormonal stimulation [8].

Artificial propagation before spawning season

Fish were divided into four groups: two experimental (J1-fish cultured in ponds, J2- fish raised in controlled conditions) and two control groups (JK1, JK2) distinguished from both experimental groups. To hormonally stimulate the fish from experimental group’s ovopel was used. One pellet (average weight about 25 mg) contains a mammalian GnRH analogue (D-Ala6, Pro9 Net-mGnRH at dose 18-20 ug) and opamine antagonist: metoclopramide (dose 8-10 mg) [3]. Ovopel pellets were pulverized in a mortar and then dissolved in 0.9% NaCl. Before manipulations fish were anaesthetized in a solution of -phenoxyethanol (0.5 mgÌdm-3). Females received double injection. Priming at dose of 0.1 pellet kg-1 and resolving one after 12 hours at dose of 1 pellet kg-1. The males were treated with one dose of 0.5 pellet kg -1. Fish from the control groups were injected with 0.9% NaCl. Injections were intraperitoneal at the ba se of pelvic fin [24].

Collection of gametes and incubation

Ripe gamete donors were anesthetized in a solution of 2-phenoxyethanol (0.5 ml per dm-3). Milt was collected with plastic syringes of calibration to 0.01ml and kept at 4 °C until further treatment. Motility of sperm was determined by observation under the microscope (500 x magnitudes) after diluting the semen just before the observation in 0.5% solution of NaCl in the ratio of 1:1 [9].

Between the 24 and 40th hour from the resolving injections the ripeness of female’s gonads was controlled every hour. Oocytes were collected to plastic containers. The eggs were fertilized using the “dry method” [10]. Then was placed and incubated in Weiss jars. The incubation temperature was 14°C. When the pigmentation of eyes occurred, the percentage of embryos survival was calculated. The spawners were being kept during two weeks period after the spawning, and after that, the percentage of their survival was determined.

Statistical analysis

Statistical differences between groups (spermiation success and incubation success) were analyzed with Duncan's multiple range test (p < 0.05) [21].


Off-seasonal reproduction of ide raised in cultured stock

The milt was collected from the males stimulated hormonally as well as from those in the control group. Volumes of the semen obtained in both groups were small and didn’t differ significantly between each other. In comparison to the control group, the semen of males stimulated hormonally had higher spermatozoa mobility (Table 1).

Table 1. The results for off-seasonal reproduction of ide (Leuciscus idus) from the cultured stock

Group of fish



Nos. of males



Spermiation success (%)



Quantity of milt (ml·kg-1)

1.5 + 0.2

1.7 + 0.2

Spermatozoa motility (%)



Nos. of females



Percentage of ovulation



The latency time (hrs)



Embryos survival (%)


84.1 – 95.2

Spawners survival (%)



All females after hormonal stimulation ovulated. The latency time was relatively long and exceeded 30 hours. Survival of embryos to the eyed-egg-stage was reaching 95% (Table 1). In the control group, during the observation after the experiment, mortality of spawners was noticed.

Off-seasonal reproduction of ide raised in controlled conditions

The volume and quality of milt was close to the results of spawning of the fish from the cultured stock (Table 2). The spermatozoa motility did not exceed 65%. Males raised in captivity were delivering the semen almost all the time.

Table 2. The results for off-seasonal reproduction of ide (Leuciscus idus) raised under controlled conditions

Group of fish



Nos. of males



Spermiation success (%)



Quantity of milt (ml·kg-1)

1.1 + 0.2

1.3 + 0.2

Spermatozoa motility (%)



Nos. of females



Percentage of ovulation



The latency time (hrs)



Embryos survival (%)

48.2 + 2.1 a

68.1 + 2.3 b

Spawners survival (%)



After off-season hormonal stimulation some of reproductive females did not deliver gametes, and in the control group one of the females ovulated. Time between the injection and ovulation exceeded 30 hours. Survival of embryos to the eyed-egg-stage was relatively low, and in the case of the female not stimulated hormonally, did not exceed 50% (Table 2). All spawners survived manipulations which had been carried out during the experiment.


In the study of ide reproduction, many researchers tried to determine the most optimal hormonal treatment [2, 5, 11, 12]. On their basis it has been said, that ovopel appears to be most appropriate stimulator of gametes maturation for this species, and this is why it was used in this experiment.

However, as stated by Kucharczyk et al. [12], using ovopel at artificial reproduction of ide, in relation to other hormonal agents, may cause increased mortality of spawners. The above-mentioned results do not confirm that. The difference probably results from conducting the reproduction of cultured fish, not fish captured in natural environment. The latter are being manipulated more, transported and thus subjected to injuries. These factors and the stress connected with them cause greater mortality before and after the artificial spawning. Results of this experiment, however, showed that there were no losses after spawning of the fish being stocked in controlled condition.

Like in the case of fish caught in their natural environment [2, 12,13] no difficulties in obtaining gametes from ide females were noticed. Nearly all females stimulated hormonally delivered them. The interesting fact is ovulation of one of the females that had not been treated with hormones. This kind of event has not been reported before in papers on off seasonal ide reproduction. It can be explained by the probability that the fish could came from the stock which had been raised in controlled conditions, kept in environmental conditions close to optimal for this species. It has also been used to small tanks and to the presence of people, which caused a decrease in its stress reactions and reaching the gametes maturation in natural way.

During the artificial reproduction of cyprinid fishes, especially those coming from natural populations, getting the eggs is usually not a problem but getting the sufficient amount and quality of the milt [12, 14]. These parameters improve for many species after applying the hormonal stimulation [19, 22]. Hormonal injections do not influence the volume of ejaculate rarely, and only at fish of smaller sizes [7, 18, 23]. In research concerning about out off season ide reproduction similar problems were examined [11, 14]. Authors focus on necessity of hormonal stimulation of males captured in their natural environment. In this experiment the percentage of males giving soft roe in experimental and control groups was the same. Hormonal stimulation had no significant influence on amount of semen produced by males; however, it had influenced its quality. Motility of spermatozoa increased thus influencing the percentage of fertilization. This confirms the results obtained by Kucharczyk et al. [12].

Biological value of gametes of fish cultured in controlled conditions, expressed by survival of embryos until the eyed-egg-stage, was significantly lower than in the second experimental group. However, it was not very different from the results obtained in the case of reproduction of ide from the wild stock given in other studies [2, 12].


  1. There is possible to do out-off-season spawning of ide under controlled conditions.

  2. There were no differences in the spawning results of different cultured stocks, except numbers of gametes.

  3. The obtained results in out-off-season spawning of ide were similar to results usually obtain in the spawning season.


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Katarzyna Targońska-Dietrich, Tomasz Zielazny,
Dariusz Kucharczyk, Andrzej Mamcarz, Roman Kujawa
Department of Lake and River Fisheries
University of Warmia and Mazury in Olsztyn
M. Oczapowskiego5, 10-718 Olsztyn-Kortowo, Poland
e-mail: koszo@post.pl

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