Does stress make males? An experiment on the role of glucocorticoids in anuran sex reversal.

Environmentally sensitive sex determination may help organisms adapt to environmental change but also makes them vulnerable to anthropogenic stressors, with diverse consequences for population dynamics and evolution. The mechanisms translating environmental stimuli to sex are controversial: although several fish experiments supported the mediator role of glucocorticoid hormones, results on some reptiles challenged it. We tested this hypothesis in amphibians by investigating the effect of corticosterone on sex determination in agile frogs (Rana dalmatina). This species is liable to environmental sex reversal whereby genetic females develop into phenotypic males. After exposing tadpoles during sex determination to waterborne corticosterone, the proportion of genetic females with testes or ovotestes increased from 11% to up to 32% at 3 out of 4 concentrations. These differences were not statistically significant except for the group treated with 10 nM corticosterone, and there was no monotonous dose-effect relationship. These findings suggest that corticosterone is unlikely to mediate sex reversal in frogs. Unexpectedly, animals originating from urban habitats had higher sex-reversal and corticosterone-release rates, reduced body mass and development speed, and lower survival compared to individuals collected from woodland habitats. Thus, anthropogenic environments may affect both sex and fitness, and the underlying mechanisms may vary across ectothermic vertebrates.

Review of Central-Eastern European Propagation and Larvae Nursing Method for Common Carp (Cyprinus carpio L.).

Common carp (Cyprinus carpio L.) as a cultivated fish species has huge importance all over the world. According to FAO statistics, carp is the third most widely bred freshwater pond fish species; only two other Cyprinids (silver carp and grass carp) are bred in higher amounts. Carp is native all over Asia and in a large part of Europe. As a result of human intervention, at present, carp are widespread all over the world, except for the Arctic region. Carp breeding was launched in the antique period, in the ancient Chinese Empire and the Roman Empire. The presently applied method of breeding of common carp has a long evolution. From the effectiveness point of view, the propagation and early-life nursing are crucial parts of carp production, as they provide seed stocks for the further growing section. Without effective propagation, there is no intensive carp production. Nowadays, more advanced propagation methods are available all over the world; however, in the current review, only the main milestones and production efficiency of the propagation and nursing method used in the ponds of Eastern Central Europe are discussed. In the historical overview of carp reproduction, first the natural reproduction, then the semi-extensive and intensive hatchery propagation are presented and investigated in detail. The analysis focuses on the advantages and disadvantages of the method. In particular, the different important milestones of the advanced hatchery method are shown and explained. The effectiveness is proven even with practical calculations. Not only the reproduction, but the pond nursing method is also presented and discussed, concentrating on the management of evolutionarily adapted natural feeds (Zooplankton) and their effect on the survival of fish larvae.

Carp Breeding in the Carpathian Basin with a Sustainable Utilization of Renewable Natural Resources.

In the Central European region, there is a long tradition of breeding fish in artificially constructed ponds. As the area belongs to the temperate zone, farmed fish need to survive cold winter periods. Common carp (Cyprinus carpio L.), which is an omnivorous, bioturbating species, is well adapted to warm and cold periods and the alluvial water environment. Since the Middle Ages, a large scale, efficient carp farming methodology has been developed in the region, where production is based on natural resources (protein and fatty acid sources) of renewable water ecosystems. This summary aims to present this well-developed breedi:ng method through discussing aspects of hydrobiology and energy transfer through the food chain as well. Capabilities and effects of agro-technical treatments such as liming and organic manuring, zooplankton management and possible supplementary feedings are also reviewed. Analysing chemical processes of waters uncovers that biological production has no carbon footprint; no carbon dioxide is released into the atmosphere. In contrast, gaseous carbon dioxide diffuses into pond water containing calcium and/or magnesium, then it accumulates in algae production and, through energy migration to upper trophic levels, increases carp production. Thus, it can be declared that pond-farmed carp provides an environmentally friendly, delicious meat among products of animal origin.

Novel genetic sex markers reveal high frequency of sex reversal in wild populations of the agile frog (Rana dalmatina) associated with anthropogenic land use.

Populations of ectothermic vertebrates are vulnerable to environmental pollution and climate change because certain chemicals and extreme temperatures can cause sex reversal during early ontogeny (i.e. genetically female individuals develop male phenotype or vice versa), which may distort population sex ratios. However, we have troublingly little information on sex reversals in natural populations, due to unavailability of genetic sex markers. Here, we developed a genetic sexing method based on sex-linked single nucleotide polymorphism loci to study the prevalence and fitness consequences of sex reversal in agile frogs (Rana dalmatina). Out of 125 juveniles raised in laboratory without exposure to sex-reversing stimuli, 6 showed male phenotype but female genotype according to our markers. These individuals exhibited several signs of poor physiological condition, suggesting stress-induced sex reversal and inferior fitness prospects. Among 162 adults from 11 wild populations in North-Central Hungary, 20% of phenotypic males had female genotype according to our markers. These individuals occurred more frequently in areas of anthropogenic land use; this association was attributable to agriculture and less strongly to urban land use. Female-to-male sex-reversed adults had similar body mass as normal males. We recorded no events of male-to-female sex reversal either in the laboratory or in the wild. These results support recent suspicions that sex reversal is widespread in nature, and suggest that human-induced environmental changes may contribute to its pervasiveness. Furthermore, our findings indicate that sex reversal is associated with stress and poor health in early life, but sex-reversed individuals surviving to adulthood may participate in breeding.

Development of sperm vitrification protocols for two endangered salmonid species: the Adriatic grayling, Thymallus thymallus, and the marble trout, Salmo marmoratus.

Vitrification was applied to the sperm of two endangered fish species of Soca River basin in Slovenia, the Adriatic grayling (Thymallus thymallus) and marble trout (Salmo marmoratus) following testing different cooling devices and vitrifying media. Sperm was collected, diluted in species-specific non-activating media containing cryoprotectants, and vitrified by plunging directly into liquid nitrogen without pre-cooling. Progressive motility, curvilinear velocity, and straightness of fresh and vitrified-warmed sperm were evaluated with computer-assisted sperm analysis (CASA). Fertilization trials were carried out to test the effectiveness of vitrification in the case of grayling. A protocol utilizing a glucose-based extender, 30% cryoprotectants (15% methanol + 15% propylene glycol), 1:1 dilution ratio, and droplets of 2 µl on a Cryotop as cooling device yielded the highest post-thaw motility values for both Adriatic grayling (7.5 ± 6.5%) and marble trout (26.6 ± 15.8%). Viable embryos were produced by fertilizing eggs with vitrified grayling sperm (hatching 13.1 ± 11.7%, control hatching 73.9 ± 10.4%). The vitrification protocol developed in this study can be utilized in the conservation efforts for the two species as an alternative to slow-rate freezing when working in field conditions or when specific equipment necessary for slow-rate freezing is not available.

Haemangioma in the oesophagus of a red-eared slider (Trachemys scripta elegans).

A haemangioma developing in the wall of the oesophagus and protruding into its cavity is reported for the first time from a Red-eared Slider (Trachemys scripta elegans). As the tumour mechanically hampered swallowing, the animal was unable to eat and consequently developed a poor condition. Histopathology of the tumour revealed all characteristics of a haemangioma: the blood-filled blood-vessels having an irregular cross-section were lined with endothelial cells. Claudin-5 immunohistochemical antibodies were employed for characterising the tumour, and this examination confirmed our initial diagnosis of a haemangioma.