Application of high hydrostatic pressure (HHP) shock to induce triploid development in the European grayling (Thymallus thymallus L.).

Use of parental individuals from allochthonous lineages to produce stocking specimens resulted in genetic contamination of many local European grayling (Thymallus thymallus) populations. Stocking programs including triploid individuals that are not able to reproduce with stocks located in natural habitats due to being reproductively sterile could contribute to populations of the European grayling and prevent regional lineages from the genetic pollution. The main goal of the present research, therefore, was to provide conditions for triploidization of the European grayling and examine gonads of triploid individuals. High Hydrostatic Pressure (HHP) shock (9000 psi/5 min.) applied to activated and incubated grayling eggs at 10 °C between the 17 min. 30 sec. and 20 minute timepoints subsequent to the time of insemination resulted in inhibition of the second polar body release and triploidization. Histological analysis confirmed that all 1-yr-old cytogenetically confirmed triploids had gonadal tissues indicative of sterility with the gonads being composed of connective tissue including fibroblasts, adipocytes and degenerated epithelial structures without there being any differentiated germ cells detected. There were no differences in the body length and Fulton's condition factor between 1 year old diploid and triploid grayling, however, the average body weight was markedly less in triploid than diploid specimens. Although, 1-yr-old triploid European female and male graylings did not have fully developed gonads, before these triploid specimens are deemed safe, as a result of being reproductively sterile for stocking in natural grayling habitats, there needs to be further examination of the 2- and 3-year-old triploid specimens.

A Comparison of Pneumatic and Hand Stripping of Whitefish (Coregonus lavaretus) Eggs for Artificial Reproduction.

We describe the technique of pneumatic stripping of whitefish (Coregonus lavaretus) eggs with the use of oxygen, nitrogen, and air. Eggs obtained via the traditional method (by pressing the abdominal surfaces) served as a control group. It was established that the gas flow rate during pneumatic stripping should not exceed 0.5 L∙min-1, since higher air flow resulted in increased post-spawning mortality. The pneumatic stripping method of egg collection was no faster than hand stripping; however, the time required per female was more consistent. It was found that the pH of the ovarian fluid obtained during hand and pneumatic stripping was not related to the success rate of fertilization. Pneumatic stripping resulted in a higher quality of collected eggs and a higher and more consistent hatching rate as compared with the hand-stripped samples, regardless of the gas used. The results presented here lead us to recommend the pneumatic method for obtaining eggs from whitefish, since it is a simple, reproducible method and improves the reproductive performance and developmental success of the fish eggs.

Effectiveness of the Air Stripping in Two Salmonid Fish, Rainbow Trout (Oncorhynchus Mykiss) and Brown Trout (Salmo Trutta Morpha fario).

Egg collection is one of the most crucial procedures during fish reproduction in salmonid hatcheries. Classic methods involve the use of hand massage on fish abdomens to expel the eggs. An alternative method uses the pressure of gas injected into the body cavity, which causes the subsequent release of the eggs. This method is believed to have less negative effects on both the welfare and egg quality of the broodstocks. Herein, we compare the results of air and hand stripping methods with respect to one-year survival and egg quantity and quality in two salmonid fish, rainbow trout (Oncorhynchus mykiss) and brown trout (Salmo trutta morpha fario). Our results indicate that air stripping yielded a better quality of eggs and higher one-year survival rate in rainbow trout. In addition, air stripping resulted in lower mortality rate than the group subjected to hand stripping (25% vs. 35%). The pH and hatching rate of the hand stripped group was lower than those of the air stripped group. In the case of brown trout, the quality of eggs obtained by both hand and air-stripping methods was similar; however, the one-year losses in fish were higher in air stripped group (15% compared to 0% in hand stripped fish). Although the advantages of air stripping method over hand stripping in terms of egg quality might not be observed in all salmonid species, the air-stripping procedure might be a promising option to be adopted in hatcheries as it ensures a high level of reproducibility and efficiency.

Evaluation of oxidative stress markers in the heart and liver of rainbow trout (Oncorhynchus mykiss walbaum) exposed to the formalin.

The aim of this study was to examine change in lipid and protein oxidation biomarkers, transamination enzymes and lactate dehydrogenase activities, lactate and pyruvate levels in liver and heart tissue of rainbow trout (Oncorhynchus mykiss Walbaum) that was exposed to formalin baths. Increase of 2-thiobarbituric acid reactive substances and carbonyl derivatives of protein oxidative destruction was noticed only in cardiac tissue of formalin-exposed fish. Activity of lactate dehydrogenase and lactate level in the cardiac tissue were elevated, indicating active glycolysis. Effects of formalin disinfection were different in both tissues. Aldehydic and ketonic derivatives of oxidatively modified proteins in liver were consistently reduced upon exposure to the formalin. In support of this, decrease in alanine and aspartate aminotransferases was noticed. Formalin disinfection of rainbow trout results in metabolic plasticity, predominantly in liver with decreased levels of oxidative stress biomarkers and aminotransferases activity. Formalin-induced oxidative stress in the cardiac tissue was more considerable.

Tissue-specific responses of oxidative stress biomarkers and antioxidant defenses in rainbow trout Oncorhynchus mykiss during a vaccination against furunculosis.

The present study was conducted to evaluate the effects of vaccination against furunculosis on responses of oxidative stress and antioxidant defenses in rainbow trout Oncorhynchus mykiss muscle, gills, liver, and brain tissues. The oxidative stress markers (malondialdehyde and carbonyl derivatives of protein oxidative destruction levels), antioxidant defenses (superoxide dismutase, catalase, glutathione reductase, and glutathione peroxidase), and total antioxidant capacity in different tissues of rainbow trout were measured. Our data showed that exposure of trout to vaccine against furunculosis produced changes (either increase or decrease) in oxidative stress and antioxidant enzymes responses, and these responses showed marked organ differences, associated with tissue patterns. Our study demonstrated that vaccinated trout showed alteration in antioxidant defenses and oxidative stress responses, with higher severity in the liver, compared with other tissues. Our data also suggest that vaccination against furunculosis induced lipid peroxidation in gill and liver tissues. However, muscle and brain tissue are capable of restoring its pro- and antioxidant balance after vaccination.

Influence of deltamethrin on nonspecific cellular and humoral defense mechanisms in rainbow trout (Oncorhynchus mykiss).

The influence of deltamethrin on the innate immunity in rainbow trout was examined. Fish were immersed in deltamethrin at doses of 1, 2, and 4 microg/L for 30 min. The results showed that deltamethrin at doses of 2 and 4 microg/L decreased phagocytic activity of spleen macrophages and proliferative response of pronephros lymphocytes at days 1, 2, and 5 after immersion. Deltamethrin at these doses decreased the lysozyme activity, total protein, and immunoglobulin levels in serum. The greatest immunosuppressive influence of deltamethrin at dose 4 microg/L was observed at the end of the study.