A minimally invasive procedure for blood extraction from Xenopus laevis allows follow up studies without euthanasia.

Blood extraction is extremely important for the development of scientific research; however, the existing methods for amphibian´s blood sampling are invasive, mainly leading to the euthanasia of the animal. Therefore, less intrusive methods that allow the obtention of multiple samples from the same individual, are needed as an alternative to the common methods available. Hence, the aim of this study was to propose a minimally invasive method for obtaining blood from the hind leg of Xenopus laevis, that allows continuous sampling without compromising the wellbeing of the organisms. With this method, it was possible to extract blood and plasma from adults and juveniles, and the amount of sample was enough to perform biochemical and molecular assays to assess the viability of the blood. The results also revealed that this method is a convenient alternative to obtain blood without affecting the welfare of the experimental organisms, avoiding the cull of the animals, and the samples are viable for their use in follow up studies.

Simultaneous nitrification/denitrification and desulfurization of wastewater polluted with ammonium, COD and sulfide: effectiveness of a new up-flow vertical hybrid reactor.

Industrial wastewater discharges pose an environmental risk. Here, the effectiveness of an up-flow vertical hybrid system, operating with synthetic and industrial wastewater was investigated, as a new approach to perform nitrification/denitrification and desulfurization within a single reactor. The hybrid reactor is divided in two reaction zones, the oxic and anoxic. The removal of chemical oxygen demand (COD), ammonium, and sulfide was investigated, highlighting changes in microbial diversity. The reactor was evaluated at hydraulic residence time (HRT) of 1.6 days, and its performance throughout 180 days is presented in four stages. In stages I-II, high COD and ammonium removal was obtained with synthetic wastewater. In stage-III, sulfide-rich synthetic wastewater did not alter the system, attaining COD, ammonium, and sulfide removal efficiencies of 81, 99.5, and 99.7%, respectively. In the last stage, a mixture of effluents was fed into the reactor at loading rates of 277 mg COD/L-d, 46.5 mg NH4 +-N /L-d, and 15 mg HS--S /L-d. Sulfide and ammonium removals were 100% and 99.9%, respectively. However, low COD removal was observed, being of 51%, and the system removed 97% in terms of BOD5. The structure and microbial diversity also changed. Sulfide feeding, induced the proliferation of sulfur oxidizers like Thiomiscropira and Thiobacillus. Industrial wastewater enhanced the abundance of Pseudomonas (15.53%) and favored the proliferation of new bacteria of the genus Truepera (2.98%) and Alicyclipilus (7.56%). This is the first study reporting simultaneous nitrification/denitrification and desulfurization to remove ammonium, COD and sulfide from complex industrial wastewater using an up-flow vertical hybrid reactor.

Chronic stress of rainbow trout Oncorhynchus mykiss at high altitude: a field study.

The stress response of Oncorhynchus mykiss in high-altitude farms in central Mexico was investigated over two seasons: the cool (9·1-13·7° C) dry winter season, and the warmer (14·7-15·9° C), wetter summer season. Fish were subjected to an acute stress test followed by sampling of six physiological variables: blood cortisol, glucose, lactate, total antioxidant capacity, haemoglobin concentration and per cent packed cell volume (VPC %). Multivariate analyses revealed that lactate and total antioxidant capacity were significantly higher in the summer, when water temperatures were warmer and moderate hypoxia (4·9-5·3 mg l(-1) ) prevailed. In contrast, plasma cortisol was significantly higher in the winter (mean ± s.e.: 76·7 ± 4·0 ng ml(-1) ) when temperatures were cooler and dissolved oxygen levels higher (6·05-7·9 mg l(-1) ), than in the summer (22·7 ± 3·8 ng ml(-1) ). Haemoglobin concentrations (mg dl(-1) ) were not significantly different between seasons, but VPC % was significantly higher in the summer (50%) than in the winter (35%). These results suggest that in summer, effects of high altitude on farmed fish are exacerbated by stresses of high temperatures and hypoxia, resulting in higher blood lactate, increased total antioxidant capacity and elevated VPC % levels.

Boldness and aggressiveness in early and late hatched three-spined sticklebacks Gasterosteus aculeatus.

Levels of boldness and the degree of aggressiveness were compared in juvenile three-spined sticklebacks Gasterosteus aculeatus that had hatched early and late in the breeding season. The most striking result found in this study was that early hatched individuals were bolder when exploring a novel environment than were late-hatched individuals. No differences in levels of aggression between early and late hatchlings were found, but a relationship between boldness and aggressiveness was present regardless of hatching date. The implications of these findings are discussed in the light of research on individual variation in behaviour and the development of behavioural syndromes.

Consistent boldness behaviour in early emerging fry of domesticated Atlantic salmon (Salmo salar): Decoupling of behavioural and physiological traits of the proactive stress coping style.

Individual variation in the way animals cope with stressors has been documented in a number of animal groups. In general, two distinct sets of behavioural and physiological responses to stress have been described: the proactive and the reactive coping styles. Some characteristics of stress coping style seem to be coupled to the time to emerge of fry from spawning redds in natural populations of salmonid fishes. In the present study, behavioural and physiological traits of stress coping styles were compared two and five months after emergence in farmed Atlantic salmon (Salmo salar), using individuals with an early or late time to emerge. Initially, compared to late emerging individuals, early emerging individuals showed a shorter time to resume feeding after transfer to rearing in isolation. Resumption of feeding after isolation was suggested to be related to boldness behaviour, rather than hunger, in the present study. This observation was repeated five months after emergence, demonstrating behavioural consistency over time in this trait. However, in other traits of proactive and reactive stress coping styles, such as social status, resting metabolism or post stress cortisol concentrations, early and late emerging individuals did not differ. Therefore, this study demonstrates that boldness in a novel environment is uncoupled from other traits of the proactive and reactive stress coping styles in farmed salmonids. It is possible that this decoupling is caused by the low competitive environment in which fish were reared. In natural populations of salmonids, however, the higher selection pressure at emergence could select for early emerging individuals with a proactive coping style.

Three-spined sticklebacks Gasterosteus aculeatus as a model for exploring behavioural biology.

Niko Tinbergen chose the three-spined stickleback Gasterosteus aculeatus for his classic behavioural studies because they are small, robust, common and easy to house in the laboratory under reasonably natural conditions and also because their behaviour, while sufficiently simple to be tractable, is still sufficiently complex to be interesting. An analysis of citation records shows that this was an inspired choice. Research on these small fish has addressed all four of Tinbergen's famous questions (causation, development, functions and evolution) and has contributed to the understanding of many different behavioural systems. The G. aculeatus literature is used here to explore several themes in fundamental behavioural biology (diet choice, shoaling, behavioural syndromes and sexual signalling) and the extent to which research using G. aculeatus has informed both fundamental and applied behavioural biology, the latter in the context of aquaculture research.