Ewa Sokołowska

Age and growth of the round goby Neogobius melanostomus in the Gulf of Gdańsk several years after invasion. Is the Baltic Sea a new Promised Land

The ages of 8 to 23·5 cm total length (L(T)) round goby Neogobius melanostomus collected monthly during 2006 and 2007 in the Gulf of Gdańsk (Baltic Sea) ranged from 2 to 6 years, with age class 4+ years dominant. Males were larger at age than females. The fastest growth occurred in the first 2 years of life in both sexes. Females were heavier at a given L(T) than males, but only for fish > c. 15 cm. A strong relationship between N. melanostomus otolith size and fish size was found, with no difference between males and females, and a significant relationship between fish growth rate and otolith growth rate, which enabled backcalculation of growth rates. Marginal increment width analysis confirmed the periodicity of annual ring formation in otoliths and showed that the most intense opaque zone formation occurs in July to August, while hyaline zone formation starts as early as September to October. It was concluded that the N. melanostomus that have colonized the southern Baltic Sea exhibit the largest size and longest life span ever recorded for this species.

Changes in brain arginine vasotocin, isotocin, plasma 11-ketotestosterone and cortisol in round goby, Neogobius melanostomus, males subjected to overcrowding stress during the breeding season.

In natural spawning grounds, breeding round goby, Neogobius melanostomus, males are exposed to various social stimuli, including high density of same-sex competitors and separation from females. We hypothesize that breeding males subjected to overcrowding in the wild experience high stress that affects their socio-sexual behavior and their relationships among conspecifics. We designed an experiment to mimic natural stimulation when highly aggregated breeding males are subjected to same-sex opponents. Males were sampled sequentially from experimental tank stocked at decreasing fish densities of 15 fish/m(2), 9 fish/m(2) and 4 fish/m(2). We studied the effects of overcrowding on male behavior and selected hormones, brain arginine vasotocin (AVT) and isotocin (IT) and plasma 11-ketotestosterone (11-KT) and cortisol as these are known to play roles in reproduction and related social interactions. The highest brain AVT and plasma cortisol levels were measured in non-aggressive males kept in the overcrowded group of 15 fish/m(2). IT level was elevated in fish kept at the lower density of 9 fish/m(2), and at which the males began to display territoriality and aggression. The plasma level of 11-KT was similar in all the males. Brain AVT and IT and plasma cortisol along with behavioral observations can be applied as species-specific indicators of the well-being of round goby males.

Environmental influence on maturation and dominance relationships in the three-spined stickleback (Gasterosteus aculeatus L.): temperature competes with photoperiod for primacy

Environmental influence on maturation and dominance relationships in the three-spined stickleback (Gasterosteus aculeatus L.): temperature competes with photoperiod for primacy In this study, the influence of a combination of different photoperiods and temperatures on the final maturation and social interactions in three-spined sticklebacks was investigated. Water temperature appears to be the principal signal affecting gonadal development and breeding activity of sticklebacks in pre-spawning and spawning periods. Males can mature independently of photoperiod and a stimulatory effect of high temperature is not diminished by light deprivation. On the other hand, low temperature can inhibit the development of secondary sexual characters in males exposed to long day or constant light. In females, lighting seems to be more decisive for complete maturation and the lack of light delays the maturation rate, even in high temperature. While kept under the same conditions, males mature quicker than females. The presence of light and visual information are crucial to establish the social position of individuals in the group. In light, a rigid social hierarchy with one dominant, sexually active male is observed. In constant darkness, however, several males in the group demonstrate every sign of sexual activity.

Annual changes in brain concentration of arginine vasotocin and isotocin correspond with phases of reproductive cycle in round goby, Neogobius melanostomus

Reproductive cycle of seasonally breeding fish is synchronized with changes of photoperiod and temperature in environment. We hypothesize that arginine vasotocin (AVT) and isotocin (IT) are involved in timing and synchronization of seasonal reproductive activity in the round goby (Neogobius melanostomus). To verify this hypothesis, we examined the annual profiles of brain AVT and IT in round goby males and females in relation to their reproductive cycle. Wild round gobies were exposed to annual environmental changes in their natural habitats from where they were sampled monthly over a year. AVT and IT were measured using HPLC with fluorescence detection preceded by solid-phase extraction. This study shows seasonal variations in brain AVT and IT levels. Profiles of changes were similar in males and females: the peak of AVT was observed before spawning in March-April, whereas that of IT during spawning in May–June. Furthermore, the lowest AVT level was noted out of breeding season from November to January, while the level of IT decreased immediately at the end of the spawning. The results show that high AVT levels correlate with pre-spawning period whereas the highest IT levels correspond to spawning. A significant decline in AVT and IT in non-spawning season coincided with the quiescent phase of gametogenesis in both sexes.

The time enzyme in melatonin biosynthesis in fish: Day/night expressions of three aralkylamine N-acetyltransferase genes in three-spined stickleback

In vertebrates, aralkylamine N-acetyltransferase (AANAT; EC 2.3.1.87) is a time-keeping enzyme in melatonin (Mel) biosynthesis. Uniquely in fish, there are several AANAT isozymes belonging to two AANAT subfamilies, AANAT1 and AANAT2, which are encoded by distinct genes. The different substrate preferences, kinetics and spatial expression patterns of isozymes indicate that they may have different functions. In the three-spined stickleback (Gasterosteus aculeatus), there are three genes encoding three AANAT isozymes. In this study, for the first time, the levels of aanat1a, aanat1b and aanat2 mRNAs are measured by absolute RT-qPCR in the brain, eye, skin, stomach, gut, heart and kidney collected at noon and midnight. Melatonin levels are analysed by HPLC with fluorescence detection in homogenates of the brain, eye, skin and kidney. The levels of aanats mRNAs differ significantly within and among organs. In the brain, eye, stomach and gut, there are day/night variations in aanats mRNAs levels. The highest levels of aanat1a and aanat1b mRNAs are in the eye. The extremely high expressions of these genes which are reflected in the highest Mel concentrations at this site at noon and midnight strongly suggest that the eye is an important source of the hormone in the three-spined sticklebacks. A very low level of aanat2 mRNA in all organs may suggest that AANAT1a and/or AANAT1b are principal isozymes in the three-spine sticklebacks. A presence of the isozymes of defined substrate preferences provides opportunity for control of acetylation of amines by modulation of individual aanat expression and permits the fine-tuning of indolethylamines and phenylethylamines metabolism to meet the particular needs of a given organ.

Cortisol and melatonin in the cutaneous stress response system of fish

The stress hormone cortisol, together with antioxidants, melatonin (Mel) and its biologically active metabolites, 5-methoxykynuramines, including AFMK, set up a local stress response system in mammalian skin. Our in vitro study of the European flounder (Platichthys flesus) was designed to examine whether Mel and AFMK would respond to cortisol while a glucocorticoid is added to the incubation medium. The concentrations of cortisol in the incubation medium mimic plasma cortisol levels seen in fish exposed to different types of stresses such as handling, confinement, high density, food-deprivation or air-exposure. We measured Mel and AFMK in skin explants and culture media using high-performance liquid chromatography (HPLC) with fluorescence detection. We also analysed melanosome response (dispersion/aggregation) in the explants subjected to the different treatments. Cortisol stimulated the release of Mel and AFMK from skin explants in a dose-dependent manner. Melanosome dispersion and a darkening of the skin explants were observed after incubation with cortisol. This study is the first to demonstrate the interrelationship between cortisol and Mel/AFMK in fish skin. Our data strongly suggest that the cutaneous stress response system (CSRS) is present in fish. The question remains whether Mel, AFMK or cortisol are synthetized locally in fish skin and/or transported by the bloodstream. The presence of the CSRS should be taken into account during elaboration of new indicators of fish welfare both in aquaculture and in the wild.