Paternal-effect-genes revealed through sperm cryopreservation in Perca fluviatilis.

Knowledge about paternal-effect-genes (PEGs) (genes whose expression in the progeny is influenced by paternal factors present in the sperm) in fish is very limited. To explore this issue, we used milt cryopreservation as a specific challenge test for sperm cells, thus enabling selection amidst cryo-sensitivity. We created two groups of Eurasian perch (Perca fluviatilis) as a model - eggs fertilized either with fresh (Fresh group) or cryopreserved (Cryo group) milt from the same male followed by phenotypic-transcriptomic examination of consequences of cryopreservation in obtained progeny (at larval stages). Most of the phenotypical observations were similar in both groups, except the final weight which was higher in the Cryo group. Milt cryopreservation appeared to act as a "positive selection" factor, upregulating most PEGs in the Cryo group. Transcriptomic profile of freshly hatched larvae sourced genes involved in the development of visual perception and we identified them as PEGs. Consequently, larvae from the Cryo group exhibited enhanced eyesight, potentially contributing to more efficient foraging and weight gain compared to the Fresh group. This study unveils, for the first time, the significant influence of the paternal genome on the development of the visual system in fish, highlighting pde6g, opn1lw1, and rbp4l as novel PEGs.

Impact of butyl benzyl phthalate on development of the reproductive system of European pikeperch, Sander lucioperca (L.).

The effect of butyl benzyl phthalate (BBP) on the sex differentiation process of fish is practically unknown. The experimental material of this study was juvenile European pikeperch [Sander lucioperca (L.)], which is gonochoristic, undergoes immediate sex differentiation, and has a fixed gonad differentiation period. The fish were fed a diet supplemented with BBP (during the sex differentiation phase: age 61-96 days post hatch) in the following quantities: 1.0; 2.0; 4.0; 8.0; 16.0 g BBP kg-1 feed. The control feed was a xenobiotic-free base feed. In the present experiment lasting 10 weeks, the survival and growth of fish, the histopathological changes of the fish gonads and the sex ratio were evaluated. After administration of the two highest doses of BBP, growth inhibition of the fish was observed. BBP also seriously disturbed the gonadal differentiation process of pikeperch. All analysed concentrations of BBP delayed testicular development and, at concentrations of 4.0, 8.0 and 16.0 g BBP kg-1, induction of the feminisation process was observed. The sex ratio was distinctly disrupted in groups receiving 8.0 and 16.0 g BBP kg-1.

Impact of diets with different proportions of linseed and sunflower oils on the growth, liver histology, immunological and chemical blood parameters, and proximate composition of pikeperch Sander lucioperca (L.).

The aim of the study was to determine the impact of applying different proportions of linseed (LO) and sunflower (SFO) oils in pikeperch diets on growth, histological changes in the liver, immunological and blood chemical parameters. The fish were fed isoenergetic and isoprotein feeds containing SFO (group 100SFO) or LO (group 100LO) in quantities of 67 g kg/feed, and a mixture of oils: 47 g SFO and 20 g LO kg/feed (group 70SFO/30LO) and 20 g SFO and 47 g LO kg/feed (group 30SFO/70LO). Dietary ratios of polyunsaturated fatty acids from the n-3 and n-6 series (n3/n6 index) were 0.36-2.15. Pikeperch were reared for 56 days in three replicates for each dietary treatment. Various dietary oils and ratios of n3/n6 did not impact fish growth, feed conversion ratio, viscerosomatic and hepatosomatic index, and size of the hepatocytes. Feeding the fish high quantities of LO and SO oils (groups 100LO and 100SFO) reduced the immunological response of the phagocytes and lymphocytes in the fish. Moreover, this resulted in significant differences among groups in the quantity of linolenic and linoleic acid in whole fish bodies, viscera, fillets, and livers. Various quantities of vegetable oils in the fish diets did not impact the quantity of arachidonic, eicosapentaenoic and docosahexaenoic acid in the fillets and livers. The immunological index and low quantities of linoleic acid in the fillets obtained in group 30SFO/70LO indicate that the n3/n6 dietary ratio of 1.35 was the most advantageous for feeding juvenile pikeperch feeds with vegetable oils.

Various aspects of piscine toxicology.

In opposition to toxicology of mammals piscine toxicology is closely connected with the conditions of external environment. The aquatic environment is necessary for embryonic development and after hatching during short or long-lasting larval period of most fish species. An aquatic environment is polluted by many industrial and agricultural wastes. Ammonia as a toxic and common compound in water have negative influence for aquaculture especially in intensive fish culture, recirculation system and hatchery facilities. Acute toxicity of ammonia was investigated in carp Cyprinus carpio L. and developmental stages of chub Squalius cephalus L. Changes in the peripheral blood characteristics and hemopoietic tissues of carp occurred after exposition to ammonia in acute tests and 3, 5 and 10 weeks sublethal concetration. The observed increase of the concentration of most amino acids in fish intoxicated with amonia suggests that the process reflects detoxication of ammonia which takes place both in the brain and muscles after 3 weeks of exposition. Phenol intoxication tests induced considerable unfavorable changes in the blood and dystrophic and necrobiotic lesions in tissues of fish leading to dysfunction both hemopoietic and reproductive processes.In study on fish reproduction disruptors the influence of oxygenated polycyclic hydrocarbons (17-ß-estradiol, 4,7-dihydroxyisoflavone, 1,6-dihydroxynaphthalene and 1,5-dihydroxynaphthalene) and oxygenated monocyclic hydrocarbons (phenol, 4-n-heptylphenol, 4-n-buthylphenol, 4-sec-buthylphenol; 4-tert-buthylphenol) was assessed using histopathological methods. It was established that examined oxygenated aromatic hydrocarbons both natural (17-ß-estradiol and 4,7-dihydroxyisoflavone) and synthetic can disrupt the differentiation of primary and secondary sex traits in pikeperch Sander lucioperca L. The chronic activity of these "biomimetics of estrogen" can lead to the disappearance of natural fish population. In vivo and in vitro tests were used to exam dibutyl phthalate and butyl benzyl phthalate impact on the development of the reproductive system of pikeperch. Additional as multigenerational studies are needed to clarify influence long term exposure of fish to environmental concentrations of endocrine disrupting chemicals.Hydrogen peroxide used in fish therapy is known to be toxic for sensitive species. In our work safe concentrations and exposure times was evaluated for ide Leuciscus idus L. and pike Esox lucius L. fry. The intensity of lesions in gills, skin, pseudobranch and thymus of exposed fish were connected with the time of bath.Actually anesthetics are routinely required during stressful procedures with fish, but data regarding the safety of individual anesthetics to different fish species are still few and insufficient. The influence of clove oil, MS-222 and 2-phenoxyaethanol anesthesia on fish organism was investigated in our faculty with cooperation with Research Institute of Fish Culture and Hydrobiology, Vodnany, Czech Republic.