The uptake and elimination of 20 structurally diverse tetra- to heptachlorinated biphenyls were studied in zebrafish (Danio rerio), three-spined stickleback (Gasterosteus aculeatus), and Arctic char (Salvelinus alpinus). The polychlorinated biphenyls (PCBs) were administered to the fish through food, intraperitoneal injection of peanut oil, or intraperitoneal implantation of silicone capsules. The retention of the PCBs in fish exposed through their diet was related with the substitution patterns of the compounds. Ortho-substituted congeners with no unsubstituted meta-para positions had high biomagnification potential. PCBs with low biomagnification all had adjacent vicinal hydrogens, indicating that congeners with this feature may have been metabolically eliminated. The retention characteristics of the PCBs in the diet-exposed and the injected zebrafish were similar. The pattern of congeners in Arctic char indicates that they have a lower capacity to metabolize PCBs compared to three-spined sticklebacks and zebrafish. The levels in the fish exposed to the PCBs through a silastic implant were negatively correlated with the hydrophobicity of the congeners. Most probably congener-specific release rates of the PCBs from the implants mask their retention characteristics. It is suggested that food, mimicking the natural intake route, should be used in PCB exposure studies to validate extrapolations to natural situations.

The liver-X-receptors (LXRs) act as cholesterol sensors and participate in the regulation of lipid and cholesterol metabolism. The objective of this study was to determine the role of LXR during development using the zebrafish model. By in situ hybridization we showed distinct expression of lxr in the brain and the retina in the developing and adult zebrafish. Lxr ligand activation affected the expression of genes involved in lipid metabolism in zebrafish adult brain and eye as well as in zebrafish embryos. Morpholino knock down of lxr resulted in an overall impaired lipid deposition as determined by oil red O staining particularly in the head and around the eyes, and to significantly elevated levels of both total and free cholesterol in the yolk of lxr morphant embryos. The expression of genes involved in lipid and cholesterol metabolism was also changed in the lxr morphants. Furthermore, alcian blue staining revealed malformation of the pharyngeal skeleton in the lxr morphant. Our data show that Lxr is an important component of the regulatory network governing the lipid homeostasis during zebrafish development, which in turn may support a role of Lxr for normal development of the central nervous sytem, including the retina.

Mature male goldfish were exposed to different concentrations of the natural hormone 17 beta -estradiol (El). Two methods of exposure were employed, via ingestion at 0, 1, 10 and 100 ug/g food and via the water at 0, 1 and 10 mug/l. The fish were exposed fur 74-28 days during the spawning period. The males were then paired with an artificially induced, spawning female and their sexual behaviour was observed during a 15 min period. The physiological status of the fish was also examined with respect to GSI. presence of milt and spawning tubercles and the blood plasma concentration of E-2. Despite the relatively short exposure period. exposure to physiological levels of E-2 was shown to severely affect the male goldfish reproductive behaviour and physiology. In conclusion, the results from this study and the ability to interpret the effects on this well-studied species, show that the effects of E-2, and possibly other estrogenic EDCs have severe effects at several vital levels of male goldfish reproduction. The results also suggests that the hormone E-2 can act as an endocrine disrupting chemical (EDC) in the environment.

How were fishponds introduced, farmed and spread in Scandinavia and the Baltic Region in early modern times? What was their economic, social and religious importance? Which fish species were significant and why?

This book uncovers a long, now broken, tradition that barely left traces in the written record or physical environment. Its broad and multidisciplinary scope highlights the situation from medieval times until the late nineteenth century. Besides Scandinavia and the Baltic States, insights from England are also introduced.

Several socio-cultural domains have been identified: late medieval monastic fishponds; late medieval aristocratic fishponds associated with castles and manors; seventeenth and eighteenth century ponds rectory ponds as well as urban ponds from the seventeenth century to the nineteenth century.

A wide range of animals have been reported to show kin-biased behaviours, such as reduced aggressiveness and increased food sharing among relatives. However, less is known about whether wild animals also associate with relatives under natural conditions, which is a prerequisite to facilitate kin-biased behaviours and hence kin selection. We tested, by means of microsatellite polymorphism, correlations between pair-wise relatedness and pair-wise metric distance in wild brown trout ( Salmo trutta L.) under natural conditions in two streams. Our data show that young-of-the-year as well as older trout found close together also had a higher genetic relatedness in one of the two streams, whereas no relationship was found in the other stream. Very few half and full siblings were found in the second stream and under these conditions it is unlikely that kin-biased behaviours will receive positive selection. We discuss the underlying mechanisms for the observed structure and we specifically address the issue of whether the grouping of related individuals could reflect dispersal from the same spawning redds, or if it reflects active association with relatives, possibly conferring kin-selected advantages.

Multilocus F(ST) estimates revealed a pronounced genetic structure at six microsatellite loci in brown trout Salmo trutta in Nordre Finnvikelv, with at least three breeding units that remained stable over time. Significant differences in allele frequencies were found between five sections within a 3-km range, even when no physical barriers prevented fish from migrating between sections. It is argued that geological structures may rise to patterns resembling isolation by distance. Seemingly, the most important factor causing genetic differentiation in Nordre Finnvikelv is genetic drift in small populations that are geologically subdivided by a tributary and by impassable waterfalls. Some correlation between previous behavioural observations and genetic structures were found.

The effect of kinship on growth and use of space by individually PIT-tagged 1+ brown trout was studied for 11 weeks in eight stream enclosures. Each enclosure consisted of two sections, separated by a region containing PIT-detecting antennae, which enabled us to measure use of sections by all individuals. Two types of sibling groups were tested, a single sibling group, F1, consisting of four individuals that were reared together in hatchery tank 'a' (F1(a)) plus four additional siblings of the same family but raised in hatchery tank 'b' (F1(b)), and a mixed sibling group, consisting of four F1(a) individuals plus four siblings from a second family, F2. Based on kin theory and earlier laboratory studies, we expected that growth of the F1(a) individuals in the single sibling group to be greater than that of F1(a) individuals in the mixed family sibling group, but instead we found just the opposite. The variance of growth did not differ between treatments. Nor was there a difference in time F1(a) individuals spent together when they were in mixed versus single sibling groups. We did find that F1(a) individuals changed habitat more frequently than F2 individuals in the mixed sibling group but less frequently than F1(b) in the single sibling groups. Thus, our predictions based on kin theory for growth and behavior of brown trout were not supported by our data, and we suggest that the role of kin recognition for the ecology of salmonids deserves further attention.

In mammalian and avian vertebrate groups, androgens act as controlling agents on male aggression and courtship behaviour by their conversion to oestrogens by cytochrome P450 aromatase in well-defined brain regions. Despite the fact that bony fishes have exceptionally high brain aromatase activity, little is known about it's possible regulatory effects on the reproductive behaviours of teleosts. In this study, Endler guppy males (Poecilia reticulata) were subjected to 26-29 days of 24-h exposure to two different concentrations (15 and 100 pg/L) of the aromatase inhibitor fadrozole in the water. Compared with the control males, two of three courtship activities in males exposed to the higher concentration were reduced when they were paired with receptive stimulus females. Reduction in brain aromatase activity was confirmed in both exposed groups with the use of the tritiated water assay.

Brain estrogen production, performed by the enzyme aromatase, can be disrupted/affected in teleost fish exposed to endocrine disruptors found in polluted aquatic environments. The guppy (Poecilia reticulata) was previously studied and confirmed to suffer negative effects on reproductive behaviors following inhibition of the brain aromatase reaction. Here adult guppies (Poecilia reticulata) of both genders were subjected to known endocrine disruptors: the androgen androstenedione (A), the synthetic estrogen 17 alpha-ethinylestradiol (EE2), and the estrogenic surfactant 4-nonylphenol (NP), at high (50 mu g/L) and at environmentally relevant concentrations (10 ng/L EE2, 5 mu g/L NP, and 0.7 mu g/L A) for 2 weeks followed by measurements of brain aromatase activity (bAA). In the adult males, bAA was stimulated by A and EE2 at 50 mu g/L. Female activity was also stimulated by the higher estrogenic treatment. At environmentally relevant concentrations only the EE2 treatment affected bAA, and only in males. The alkylphenolic substance NP produced no effect in either of the experiments, not on males nor females. The results indicate that short-term steroid treatments have stimulatory effects on guppy brain aromatase even at concentrations that can be found in the environment. We thus suggest bAA of adult guppies to be a suitable bioindicator of endocrine disruptors.

Understanding the neuroendocrinological mechanisms behind reproductive behaviour is fundamental when studying endocrine disruption. Neuroestrogen production is a key step in the activation of reproductive behaviours among vertebrates. The actions of estrogens are transmitted through estrogen receptors (ERs) in distinct brain nuclei. Here we report alterations in reproductive behaviours in guppy males following 55-day food treatments with the antiestrogen fulvestrant (ICI) and the synthetic oestrogen 17α-ethinylestradiol (EE2). Male courtship and mating attempts were severely reduced in EE2 treated males after both 27-30 and 41-55 days of treatment when paired with females. ICI treatment gave a significant reduction in the frequency of sigmoid display behaviour after 27-30 days of treatment, and an almost 2.5-fold increase in gonopodium thrusting after 41-55 days of treatment. ICI treated males also decreased their frequency in successful mating attempts in comparison to the control males. The neurological effects of ICI were confirmed by Real Time-PCR analysis for brain aromatase and ERα gene expression. ICI treatment suppressed aromatase expression to 64% and stimulated ERα gene expression by over 300%. These results indicate that oestrogen action via ERs may play an important role for the complete display of male courtship and mating behaviour in the guppy. The results also suggest that local steroids are involved in regulating brain aromatase expression and that the negative effects of EE2 on sexual behaviour are linked via endocrine disruption of gonadal function.

Behaviour studies are used in toxicology research as they are excellent tools to measure physiological end-points caused by exogenous chemicals. In mammals both reproductive and non-reproductive behaviours have been used for a long period of time, whereas in teleost fishes non-reproductive behaviours have received little attention compared to reproductive behaviours. Recent advances in measuring stress related behaviours in zebrafish have provided additional tools to understand behaviour toxicology in fish. One species with well documented reproductive behaviour disturbed by different toxicants is the guppy, which is better suited than zebrafish for reproductive behaviour studies and therefore might be a better model organism for comparative behaviour studies in fish toxicology. Here we report new applications for non-reproductive behaviours in guppy and test these behaviours on males treated with the endocrine disruptor 17α-ethynylestradiol at environmentally relevant concentrations. 17α-ethynylestradiol increased freezing and bottom-dwelling when fish were placed in a non-familiar aquarium, but did not significantly affect shoaling behaviour. These results are similar to the anxiogenic behaviours seen in rats treated perinatally with 17α-ethynylestradiol and add more concern to the impacts of endocrine disruptors on aquatic wildlife.

Low concentrations of psychiatric drugs are commonly found in surface waters around the world, and their ability to alter behaviors in aquatic fauna has raised concerns about ecological effects. One critical effect that contamination can have in aquatic systems is modifications of reproductive behaviors. In this study, we hypothesized that oxazepam, a common anxiolytic drug and aquatic contaminant previously shown to induce anti-anxiety behavior in salmonids, also affects the reproductive behavior of Atlantic salmon (Salmo salar). To test this hypothesis, small sexually mature Atlantic salmon males (mature parr) were exposed to concentrations of oxazepam comparable to that of effluent water (1.9 mu g/L). After 5 days of exposure, their reproductive behavior was compared with that of unexposed control individuals. Behavior was monitored in a large-scale flume aquarium that also contained a large male and a female to mimic natural spawning conditions for Atlantic salmon. Three behaviors were quantified for the mature parr; (a) amount of courting behavior towards female, (b) proximity to female, and (c) amount of attacks received from the large male. In contrast to our hypothesis, no difference in either courting behavior, proximity to female, or received aggression could be found between exposed and control male parr. Hence, our results indicate that there is no acute effect of the highest levels of oxazepam concentration reported for urban environments on spawning behavior of mature male parr, although chronic effects could not be ruled out.

In the present study, the effects of copper (CuSO4) on the ability of mature male brown trout Salmo trutta parr to detect and react both physiologically and behaviourally to female pheromones were studied. The study was composed of two parts. In the first experiment, priming effects of the female pheromone prostaglandin F-2 alpha (PGF(2 alpha)) were evaluated by determining the amount of milt produced and the blood plasma levels of 11-ketotestosterone (11-KT) and 17 alpha,20 beta-dihydroxy-4-pregnen-3-one (17,20 beta-P) after the PGF(2 alpha) exposure. In the second experiment, male parr were placed in a large stream tank together with a group of adult males and ovulated females and their individual behaviours were recorded. In the priming experiment, the amount of expressible milt was significantly lower, less than half, in groups exposed during 4 days to 10 or 100 mu g l-1 copper compared with control parr only exposed to water. No significant differences were observed in plasma levels of 11-KT and 17, 20 beta-P. During the behavioural experiment, exposed parr spent less time with the female and had a lower number of courting events. Blood plasma levels of 11-KT were, however, significantly higher in the group exposed to 100 mu g l-1 copper compared with the control group. Furthermore, the exposed group spent significantly less time swimming upstream than did the control group. The present study demonstrates that exposure to copper affects reproductive behaviours and endocrinology of S. trutta male parr.

Reproductive behaviour of brown trout (Salmo trutta L.) from an anadromous stock was Studied in a large stream water aquarium. Four adult males and two ovulated females were placed in the aquarium together with eight mature male parr. Four of the parr were exposed during the previous 4 days to two concentrations (0.1 or 1.0 mu g l(-1)) of the pyrethroid pesticide cypermethrin (a disrupter of olfactory receptor function) and four of the parr to the solvent ethanol. The behaviour of all fish was followed for 24 h and then blood and milt was collected. Exposure to the higher concentration of cypermethrin disturbed the reproductive behaviour of the parr. They displayed fewer courting events, spent less time near the nesting females and had lower volumes of strippable milt. They also had significantly lower amounts of 11-ketotestosterone (11-KT) in the blood plasma than the control roup. The higher cypermethrin group also had significantly lower levels of all these variables than the 9 lower cypermethrin group, apart from strippable milt that showed no significant differences between two groups. No significant differences in non-reproductive behaviours were observed between any of the groups. In the control fish, there were significant positive Correlations between (a) the number of courting events and the amount of time spent near the female, (b) blood plasma levels of 17 alpha,20 beta-dihydroxy-4-pregnen-3-one (17,20 beta-P) and time spent near the female and (c) plasma levels of 17,20 beta-P and the number of courting events. Further, in control fish, higher plasma levels of 17,20 beta-P were observed in part-interacting with a female compared to those with no female contacts. A priming experiment confirmed a previous study that cypermethrin damages olfactory reception. Parr exposed to cypermethrin had significantly lower blood plasma levels of 17,20 beta-P and 11-KT than control males after exposure to ovarian fluid and urine (known to contain reproductive priming pheromones). When ethanol-exposed males were exposed to ovarian fluid and urine they had significantly higher plasma levels of 17,20 beta-P compared to those exposed to water only.

Although once popular prior to the last century, the aquaculture of crucian carp Carassius carassius (L. 1758) in Sweden gradually fell from favour. This is the first genetic comparison of crucian carp from historic man-made ponds in the Scandinavian Peninsula. The aim was to identify old populations without admixture and to compare the relationship of pond populations from different provinces in Sweden. In total, nine microsatellite loci from 234 individuals from 20 locations in varied parts of Sweden were analysed. The genetic distances of crucian carp populations indicated that the populations in the southernmost province of Sweden, Scania, shared a common history. A pond population in the province Småland also showed a common inheritance with this group. In the province Uppland, further north in Sweden, the population genetic distances suggested a much more complex history of crucian carp distributions in the ponds. The data showed that there are some ponds with potentially old populations without admixture, but also that several ponds might have been stocked with fish from many sources.

Hybridization plays a pivotal role in evolution, influencing local adaptation and speciation. However, it can also reduce biodiversity, which is especially damaging when native and non-native species meet. Hybridization can threaten native species via competition (with vigorous hybrids), reproductive resource wastage and gene introgression. The latter, in particular, could result in increased fitness in invasive species, decreased fitness of natives and compromise reintroduction or recovery conservation practices. In this study, we use a combination of RAD sequencing and microsatellites for a range-wide sample set of 1366 fish to evaluate the potential for hybridization and introgression between native crucian carp (Carassius carassius) and three non-native taxa (Carassius auratus auratus, Carassius auratus gibelio and Cyprinus carpio) in European water bodies. We found hybridization between native and non-native taxa in 82% of populations with non-natives present, highlighting the potential for substantial ecological impacts from hybrids on crucian carp populations. However, despite such high rates of hybridization, we could find no evidence of introgression between these taxa. The presence of triploid backcrosses in at least two populations suggests that the lack of introgression among these taxa is likely due to meiotic dysfunction in hybrids, leading to the production of polyploid offspring which are unable to reproduce sexually. This result is promising for crucian reintroduction programs, as it implies limited risk to the genetic integrity of source populations. Future research should investigate the reproductive potential of triploid hybrids and the ecological pressures hybrids impose on C. carassius.

The conservation of threatened species must be underpinned by phylogeographic knowledge. This need is epitomised by the freshwater fish Carassius carassius, which is in decline across much of its European range. Restriction site associated DNA sequencing (RADseq) is increasingly used for such applications, however RADseq is expensive, and limitations on sample number must be weighed against the benefit of large numbers of markers. This trade-off has previously been examined using simulation studies, however, empirical comparisons between these markers, especially in a phylogeographic context, are lacking. Here, we compare the results from microsatellites and RADseq for the phylogeography of C. carassius to test whether it is more advantageous to genotype fewer markers (microsatellites) in many samples, or many markers (SNPs) in fewer samples. These datasets, along with data from the mitochondrial cytochrome b gene, agree on broad phylogeographic patterns; showing the existence of two previously unidentified C. carassius lineages in Europe; one found throughout northern and central-eastern European drainages, and a second almost exclusively confined to the Danubian catchment. These lineages have been isolated for approximately 2.15 M years, and should be considered separate conservation units. RADseq recovered finer population structure and stronger patterns of IBD than microsatellites, despite including only 17.6% of samples (38% of populations and 52% of samples per population). RADseq was also used along with Approximate Bayesian Computation to show that the postglacial colonisation routes of C. carassius differ from the general patterns of freshwater fish in Europe, likely as a result of their distinctive ecology.

Selective serotonin reuptake inhibitors (SSRIs) are psychotropic pharmaceuticals used as antidepressants. SSRIs are commonly found in surface waters in populated areas across the globe. They exert their efect by blocking the serotonin re-uptake transporter in the presynaptic nerve ending. The present study examined whether behavioural efects to exposure to SSRI citalopram depend on personality and sex in the stickleback (Gasterosteus aculeatus). Three aspects of stickleback behaviour are examined: feeding behaviour, aggression, and boldness. We exposed sticklebacks to 350–380 ng/l citalopram for 3 weeks. Feeding and aggressive behaviour were recorded before and after exposure, whereas scototaxis behaviour was tested after exposure. The results show treatment efects in feeding and aggressive behaviour. Feeding is suppressed only in the male group (χ2=20.4, P<0.001) but not in the females (χ2=0.91, P=0.339). Aggressive behaviour was signifcantly afected by treatment (χ2=161.9, P<0.001), sex (χ2=86.3, P<0.001), and baseline value (χ2=58.8, P<0.001). Aggressiveness was suppressed by citalopram treatment. In addition, the fsh showed no change in aggression and feeding behaviour over time regardless of sex and treatment, which indicate personality traits. Only females are afected by treatment in the scototaxis test. The exposed females spent signifcantly (χ2=5.02, P=0.050) less time in the white zone than the female controls.

Citalopram is an antidepressant drug, which acts by inhibiting the re-uptake of serotonin from the synaptic cleft into the pre-synaptic nerve ending. It is one of the most common drugs used in treatment of depression, it is highly lipophilic and frequently found in sewage treatment plant effluents and surface waters around the world. Citalopram and other selective serotonin re-uptake inhibitors have, at concentrations that occur in nature, been shown to have behavioural as well as physiological effects on fish and other animals. This study is the result of several different experiments, intended to analyse different aspects of behavioural effects of chronic citalopram exposure in fish. Our model species the three-spine stickleback is common in the entire northern hemisphere and is considered to be a good environmental sentinel species. Female three-spine sticklebacks were exposed to 0, 1.5 and 15μg/l nominal concentrations of citalopram for 21 days and subjected to the novel tank (NT) diving test. In the NT test, the fish exposed to 1.5μg/l, but not the 15μg/l fish made a significantly higher number of transitions to the upper half and stayed there for significantly longer time than the fish exposed to 0μg/l. The 15μg/l group, however, displayed a significantly lower number of freeze bouts and a shorter total freezing time. The test for locomotor activity included in the NT test showed that fish treated with 1.5 and 15μg/l displayed a significantly higher swimming activity than control fish both 5-7 and 15-17min after the start of the experiment. In the next experiment we compared fish exposed to 1.5μg/l and 0.15μg/l to pure water controls with regard to shoaling intensity and found no effect of treatment. In the final experiment the propensity of fish treated with 1.5μg/l to approach an unknown object and aggressive behaviour was investigated using the Novel Object test and a mirror test, respectively. The exposed fish ventured close to the unknown object significantly more often and stayed there for significantly longer time than unexposed fish. The aggression test yielded no statistically significant effects. It is concluded that citalopram changes the behaviour of the three-spine stickleback in a way that is likely to have ecological consequences and that it must not be considered an environmentally safe pharmaceutical.

Selective Serotonin Re-uptake Inhibitors (SSRIs) are a class of psychotropic drugs used to treat depression in both adolescents and pregnant or breast-feeding mothers as well as in the general population. Recent research on rodents points to persistent behavioural effects of pre- and perinatal exposure to SSRI which last into adulthood. To study effects of developmental exposure in fish, three-spine sticklebacks were exposed to 1.5 µg/l of the SSRI citalopram in the ambient water for 30 days, starting two days post-fertilisation. After 100 days of remediation in clean water the fish were put through an extensive test battery. Feeding behaviour was tested as the number of bites against a piece of food and found to be increased in the exposed fish. Aggression levels were measured as the number of bites against a mirror image during 10 minutes and was also found to be significantly increased in the exposed fish. Novel tank behaviour and locomotor activity was tested in an aquarium that had a horizontal line drawn half-way between the bottom and the surface. Neither the latency to the first transition to the upper half, nor the number of transitions or the total time spent in the upper half was affected by treatment. Locomotor activity was significantly reduced in the exposed fish. The light/dark preference was tested in an aquarium where the bottom and walls were black on one side and white on the other. The number of transitions to the white side was significantly reduced in the exposed fish but there was no effect on the latency to the first transition or the total time spent in the white half. The results in the current study indicate that developmental SSRI exposure causes persistent behavioural effects in fish and contribute to the existing knowledge about SSRIs as environmental pollutants.

Selective Serotonin Re-uptake Inhibitors (SSRI) are mood-altering, psychotropic drugs commonly used in the treatment of depression and other psychological illnesses. Many of them are poorly degraded in sewage treatment plants and enter the environment unaltered. In laboratory studies, they have been demonstrated to affect a wide range of behaviours in aquatic organisms. In this study we investigated the effect of a three-week exposure to 0.15 and 1.5 μg/l of the SSRI citalopram dissolved in the ambient water on the feeding behaviour in three-spine stickleback (Gasterosteus aculeatus). Feeding, measured as the number of attacks performed on a piece of frozen bloodworms during a 10-min period, was reduced by 30–40% in fish exposed to both 0.15 and 1.5 μg/l citalopram. The effects of the environmentally relevant concentration 0.15 μg/l on feeding, an important fitness characteristic, suggests that the ecological significance of environmental SSRI exposure may be pronounced.

Electrophysiological studies demonstrated that the olfactory epithelium of mature male brown trout Salmo trutta parr was acutely sensitive to F-series prostaglandins (PGFS) PGF(1alpha) and PGF(2alpha), with detection threshold concentrations of 10(-11) M. The olfactory epithelium was also sensitive to the PGF metabolite 15-ketoPGF(2alpha) (threshold 10(-8) M), but did not detect a further metabolite, 13,14,-dihydro-15-ketoPGF(2alpha). Immature brown trout did not detect any of the prostaglandins tested. Exposure of mature male brown trout parr to waterborne PGF(1alpha) and PGF(2alpha) (concentration 10(-8) M). resulted in significant increases in levels of expressible milt and the plasma concentrations of 17,20-betadihydroxy-4-pregnen-3-one, testosterone and 11-ketotestosterone. The olfactory epithelium of both immature and mature male brown trout parr was sensitive to the urine and ovarian fluid from ovulated female brown trout. Exposure of mature male brown trout parr to ovarian fluid resulted in an increase in the levels of plasma 17,20beta-dihydroxy-4-pregnen-3-one whilst exposure to urine increased the levels of expressible milt. In addition, PGF(2alpha) was found to be present within both the urine and ovarian fluid of mature female brown trout. It is suggested that the F-series prostaglandins have a role as priming pheromones in male brown trout.

Selective serotonin re-uptake inhibitors are pharmaceuticals used to treat a range of psychological disorders. They are frequently found in surface waters in populated areas. In recent years, they have been shown to affect the behaviour of various aquatic organisms in a way that can have ecological effects. In this study, we exposed zebrafish of both sexes to nominally 0.00, 0.15 and 1.50 µg L−1 Escitalopram in flow-through tanks for three weeks. Subsequently, ten swimming behaviour parameters were quantified using high-resolution video tracking. There were noticeable gender differences in the behaviour responses to Escitalopram. Female fish exposed to 1.50 µg L−1 Escitalopram had a lower maximum swimming velocity, stopped less often and exhibited increased boldness (reduced thigmotaxis) compared to controls. Male fish exposed to 1.50 µg L−1 had a lower maximum swimming velocity compared to control fish. At the end of exposures, both length and weight of the females exposed to 1.50 µg L−1 Escitalopram were significantly less than the group of control fish. In addition, males exposed to 1.50 µg L−1 Escitalopram were significantly shorter than control fish. The behaviour, weight and body length of the fish exposed to nominally 0.15 µg L−1 was not significantly different from control fish in either sex. The results of this study demonstrate that Escitalopram can affect subtle but ecologically important aspects of fish behaviour and lends further credibility to the assumption that Escitalopram is an environmentally active pharmaceutical.

The preference of juvenile Arctic char [Salvelinus alpinus (L.)] for odors from siblings and nonsiblings with different major histocompability complex class II (MHC) genotypes was studied in two-choice fluviarium tests. In the first part of the study, test fish demonstrated no preference for water scented by a sibling with a MHC genotype different from its own versus water scented by a MHC identical nonsibling. When both donors were siblings with different MHC genotypes, however, the test fish chose the water scented by the fish with the same MHC type as the test fish. The results suggest that odors with information about kinship are dependent on MHC but also on other, unknown factors. In the second part of the study, we observed that fish isolated since fertilization did not show any behavioral discrimination towards siblings, based on MHC genotype. One reasonable explanation for this result is that Arctic char learn to discriminate between odors from individuals of different MHC types.

No difference in attraction was observed in sibling Artic charr Salvelinus alpinus between water scented by dominant or subordinate major histocompability complex (MHC) identical fish observed in a two-choice fluviarium. In a second experiment, MHC identical sibling donors were given different types of food pellets before the preference test. The test fish showed a significant attraction to the sibling given the same kind of food as the test fish itself during the first 6 h of the fluviarium tests. The results suggest that diet has an influence on the odours released and can, in addition to MHC related odours, be used for information relating to group member identity.

The structure of the olfactory epithelium in developing Arctic charr, Salvelinus alpinus, was studied by light and scanning electron microscopy. Embryos, larvae, free-swimming fry, and individuals up to the age of 96 months after hatching were studied. In larvae about 10 days after hatching, microvillar and ciliated olfactory receptor cells were located in a pit and no nares were differentiated. Fifty days after hatching, two nares were present on each side of the head, and about 20 days later one or two lamellae had started to differentiate and were visible as an elevation in the caudal part of the floor of the olfactory chamber. Six months after hatching there were 5 – 10 club-shaped lamellae, which in older fish acquired a more disk-like appearance. The number and size of lamellae increased with the size of the fish, reaching the maximum number, 10 – 15 per rosette, 18 – 30 months after hatching. At 18 months, secondary folding of the lamellae had started. Eighteen months later, differentiated secondary lamellae were present and most of the central raphe was composed of indifferent epithelium. The olfactory receptors were located in the depressions between the secondary folds. The development of the olfactory organ is discussed in relation to the results of behavioural studies.

Psychoactive drugs, such as selective serotonin reuptake inhibitors (SSRI) have been identified in high levels in effluents from Swedish sewage treatment plants (STP) at concentrations high enough to give pharmacological effects in fish. In humans SSRIs are used in the treatment of depression and they have anxiolytic effects. In the present study we exposed Endler guppy (Poecilia wingei) of both sexes to citalopram that showed the highest concentrations of SSRIs in STP effluents and studied reproductive and non-reproductive behaviour. Male courting behaviours were not affected compared to control fish after 14-28 days exposure to 1μgL^-1. In two experiments exposing both sexes to 0.2, 2.3 or 15μgL^-1 for 21 days, fish exposed to the two highest doses showed anxiolytic effects when placed in a novel environment (novel tank diving test, NT). Males were only affected by exposure to 15μgL^-1. They had significantly longer latency to explore the upper half of the aquarium, more visits and longer time spent in the upper half, and showed less bottom freezing behaviour, all markers of anxiolytic behaviour. In females exposure to 2.3 or 15μgL^-1 significantly increased freezing behaviour, while no effects on other behaviour variables were observed. No effects on shoaling behaviour could be discerned. These results show that citalopram have anxiolytic effects on guppy fish and thus affect ecologically relevant behaviours of importance to survival of fish.

We compared the ability of urine and ovarian fluid from female Atlantic salmon (Salmo salar) to stimulate increase in plasma concentrations of sex steroid hormones in mature conspecific male parr (priming effect of the stimuli). We also tested the hypothesis that prostaglandin F-2 alpha (PGF(2 alpha)) may act as a priming pheromone in the tested stimulants. Individual males of salmon parr were exposed to female urine, ovarian fluid, urine-ovarian fluid mix, or PGF(2 alpha) Plasma concentrations of the sex steroids of 17,20 beta -dihydroxy-4-pregnen-3-one (17,20 beta -P) were higher in males exposed to urine, ovarian fluids, and PGF(2 alpha), compared to control males. PGF(2 alpha). and a mixture of urine and ovarian fluid also gave increased concentrations of 11-ketotestosterone (11-KT). Concentrations of PGF(2 alpha) were higher in ovarian fluids than in urine. A behavior test with mature male part in a fluviarium showed neither attraction to nor avoidance of 0.1 nM PGF(2 alpha), but plasma levels of 17,20 beta -P were significantly higher in exposed males compared to controls.

We exposed, in two successive spawning seasons, individually placed precocious male Atlantic salmon (Salmo salar) and brown trout (Salmo trutta) parr to odour stimuli (ovarian fluid and urine mix) from ovulated conspecific or heterospecific anadromous females. Atlantic salmon parr had significantly higher plasma concentrations of the hormones 17 alpha,20 beta-dihydroxy-4-pregnen-3-one (17,20 beta-P), 11-ketotestosterone (11-KT) and testosterone (T) after exposure to odours from conspecific females or from brown trout females compared to parr exposed to a control solution (0.9% NaCl). We did not observe any significant differences between the hormone levels in salmon parr exposed to the two female odours. The salmon parr exposed to conspecific odours had significantly higher volumes of strippable milt compared to the controls, but we did not find any significant differences when comparing the effect of the two female odours. Brown trout parr had significantly higher plasma 17,20 beta-P levels following exposure to heterospecific female odours compared to control males, but there was no significant difference between males exposed to the different female odours. We did not observe any significant differences in plasma levels of T and 11-KT and in milt volumes between exposed and control trout. Taken together, the results from both tested species indicate that the potency of heterospecific stimuli in stimulating increased plasma sex steroid hormone levels in male parr was as strong as stimuli from conspecific females. The results are discussed in connection to observed hybridisation between the two sympatric species.

The crucian carp (Carassius carassius) is a cyprinid fish with its natural distribution in Europe and the western part of Asia. Due to its hardiness and unique ability to survive winter anoxia, it has been translocated to small lakes and ponds, and in Northern Europe since medieval times has been used as a food source. Crucian carp was the only fish in the pond that survived anoxia. Small lakes and ponds with winter anoxia result in dense populations of stunted and slender fish. In lakes with other fish species present, the crucians' numbers and densities are low and they grow to large sizes. In the presence of piscivores such as pike, crucians are deep bodied. The presence of pike-eating crucians, or the pike odors, induce a change in the body form of crucians. The change in body form makes it more difficult for pikes to swallow crucians and the handling time increases. Closely related invasive Carassius species have become a serious threat to crucian carp populations in Central-Eastern Europe and South-Eastern England through competition for space and food resources and hybridization. The crucian carp's close relationship to goldfish (Carassius auratus), the most studied species concerning sex pheromones, has made it possible to demonstrate that sex pheromones are also present in a wild Carassius species and in their natural environment. The results indicate that two species use the same sex hormonal pheromone system. The crucian carp has become an important model for laboratory studies of olfaction and taste.

Kin recognition and discrimination are thought to occur in several species of various taxonomic groups. In salmonid fish, juveniles can discriminate between odours of siblings and nonsiblings from the same population even if the odour donors and the test fish have been reared separately since fertilization. This indicates that some genetic factor is important in the recognition process. The mechanisms behind kin recognition and discrimination have not yet been described. In the present study, we performed fluviarium tests to examine whether kin recognition and discrimination in juvenile Arctic charr are influenced by the major histocompatibility complex (MHC). Prior to the fluviarium tests, exon 2 of an MHC class II B gene in charr was analysed with denaturing gradient gel electrophoresis (DGGE) and individual genotypes were determined. In the fluviarium, when fish had the choice between water scented by an MHC identical sibling and a sibling with a different MHC genotype they preferred water from identical siblings. Moreover, water scented by an MHC different sibling was preferred to water from an MHC different nonsibling. However, we observed no discrimination when the test fish shared one allele with the nonsibling donor but no alleles with the sibling donor. Our results indicate that the MHC has a significant influence on the odours used for kin recognition and discrimination in juvenile Arctic charr.

The behavioral responses to urine and ovarian fluids from conspecific and heterospecific ovulated females were studied in mature Atlantic salmon (Salmo salar) male parr in a two-choice fluviarium. The males reacted differently to the stimulants. They spent more time in water scented by urine from salmon or brown trout (Salmo trutta L.) females compared to the time in water with ovarian fluids from salmon females. Furthermore, the males were attracted to salmon female urine (different from an indifferent reaction). Males exposed to urine of either species had higher plasma concentrations of testosterone (T) compared to unexposed controls. Measurement of the concentrations of prostaglandin F-2alpha (PGF(2alpha)) and its major metabolite 15-ketodihydroprostaglandin F-2alpha (15-ketodihydro-PGF(2alpha)) showed that the concentrations of the substances were higher in ovarian fluids of both species compared to those in urine. PGF(2alpha) showed a greater difference between ovarian fluid and urine than its major metabolite. The results suggest that urine of both species, in contrast to ovarian fluid, contain substances that attract mature Atlantic salmon male parr and that the active substances are neither PGF(2alpha) nor 15-keto-PGF(2alpha).

In the present experiment the behaviour and endocrine status of males of the brown trout, Salmo trutta L., (Salmoniformes: Salmonidae) were studied when males were kept in a stream tank with a nest digging female. Groups of mature adult males and precocious intact or anosmic male parr were placed with the nesting female so that the group resembled a natural spawning situation with big anadromous fish acting as dominant males and precocious parr acting as "sneakers". A control experiment was also run with only males without a female present. In intact parr there were significant positive correlations between the per cent of the total observation time spent with a female, milt volume, and plasma concentration of 17 alpha, 20 beta-dihydroxy-4-pregnen-3-one. Anosmic parr had significantly lower volumes of strippable milt and gonadal steroid hormone levels compared with intact parr. However, no differences were found in the control experiment. Significantly fewer anosmic parr attended and courted the nesting female and those anosmic fish that attended the female had significantly lower plasma levels of gonadal hormones. Intact parr also displayed a greater number of agonistic acts against other parr without any difference in fighting ability. No differences in aggression occured in the control experiment. In adult males together with a female, post-experimental gonadal steroid hormone levels were higher than pre-experimental levels. Positive correlations between aggression and androgen hormone levels were observed in adult males. No differences in plasma hormone levels were observed between adult males and intact precocious males. The results show that olfactory occlusion results in low steroid hormone levels and milt volumes in precocious males placed in a spawning situation. The courting behaviour was also affected by anosmia. Odours from the nesting female may have caused the enhanced plasma hormone

The behaviour of juvenile Arctic charr Salvelinus alpinus was studied in groups of four siblings composed of familiar and unfamiliar individuals or in mixed groups of four where both siblings and unrelated individuals were present. The frequency of aggressive acts was significantly higher in the mixed groups compared to the pure sibling groups and the difference was present at all levels of the hierarchy rank order, based on a dominance index, except the lowest ranked individuals. The difference was significant after but not before feeding, implying that competition with non-kin for a food resource increased the aggression. No significant difference in weight gain was observed between sibling and mixed groups during the 6 days of the experiment, but the RNA contents of lateral musculature in dominant individuals from sibling groups were significantly higher than the corresponding fish in the mixed group, suggesting a difference in growth rate when the experiments ended. No significant difference in RNA content was observed between subordinate fish of the two treatments, i.e. siblings v. mixed.

In a series of experiments, kin-biased behavior of young brown trout (Salmo trutta) was observed. The aggressiveness shown by groups of familiar siblings (siblings reared together since fertilization) and groups of unfamiliar siblings (siblings reared apart since fertilization) was significantly lower compared to that of mixed groups of two unrelated sibling groups (offspring of two different pairs of parents). The evolution of kin-biased behavior, as shown by a reduction in aggressiveness, is assumed to have evolved through a kin-selective mechanism.

Many fish are during feeding dependent on both an olfactory and gustatory sense. Olfaction that acts as the distance sense induces arousal, food search behaviour and attraction to the source, followed by examination of food items by the gustatory sense. During buccal handling the fish decide if the feed will be rejected or swallowed. Amino acids are often stimulatory to the gustatory sense and can act as feeding stimulants. There are, however, inter-species differences concerning what kinds of amino acids act as feeding stimulants or deterrents. The species differences are probably dependent on the natural food choice. As feeding stimulating molecules increase feeding and growth, but deterrents have the reverse effect, it is important to know what kind of molecules have either effect. In the present study we record mouth handling time in the omnivorous crucian carp, Carassius carassius, of agar pellets containing water extracts of meal consisting of ordinary food pellets, blue mussels or a commercial carp attractant. These tests were followed by testing with agar pellets with synthetic amino acids, based on the content of the water extracts of the food pellets that was the only feeding stimulant. Neither extracts of mussel meal or of commercial carp attractants had a stimulating effect, i.e. no significant difference in handling time compared to agar pellets with only water. A mixture of five of the major amino acids in the food pellet extract (40 mM alanine, 20 mM glycine, 20 mM arginine, 8 mM serine, 8 mM leucin) gave a significant longer handling time compared to agar pellets with only water. The handling time was also longer for the three amino acids that had the highest concentrations (40 mM Ala, 20 mM Gly, 20 mM Arg) and finally with only alanine (128 mM). Agar pellets with only Ala gave, however, a significant shorter handling time compared to agar pellets with food pellet extract. The mussel meal extract had the same content of free amino acids and their ranking order was the same as in extracts of food pellets, but at much higher concentrations. Based on the free amino acid content, the mussel extract should have stimulated feeding. This indicates that the mussel extract contained compounds that acted as feeding deterrents in omnivorous crucian carp that do not feed on blue mussels in their natural environment. Previous studies have shown that blue mussel extracts act as feeding stimulants in several bottom feeding carnivorous fish. We finally tested betaine (100 mM) but the molecule had no significant stimulating effect that has been observed in some other fish species. © 2016 The Authors

Previous studies with olfactory-disturbing pesticides resulted after exposure in disturbed behavior and physiology in fish. In the present experiment, reproductive behavior and milt volumes of precocious brown trout (Salmo trutta L.) male parr were studied in a large stream aquarium after exposure to the olfactory-disturbing fungicide 15 μg l^-1 IPBC (iodocarb; 3-iodo-2-propynyl butyl carbamate) for 96 h. The statistical analyses did not reveal any significant differences for time attending females between controls and IPBC-exposed males. Furthermore, there were no significant differences in milt volumes. However, when taking all fish into consideration, there were significant differences in milt volumes between parr that had been attending females and those had not been attending females. Controls that had attended females had significantly higher milt volumes than controls or IPBC-exposed males that had not attended females. Taking all control and IPBC parr into consideration, there was a statistically significant positive correlation between time attended females and volume of milt and gonadosomatic index (GSI), respectively. In summary, 15 μg l^-1 IPBC did not have any significant effects on mature male parr reproductive behavior and milt volumes.

Previous studies have shown kin recognition abilities in salmonid fish. Some authors have suggested that the attraction of juvenile fish to siblings may indicate preference for shoaling with kin. The aim of the present study is to test the prerequisite for the hypothesis that siblings swim spatially closer than unrelated fish during their seaward migration as smolts. Atlantic salmon (Salmo salar) eggs from three families were each reared in two tanks to create familiar and unfamiliar sibling smolts. Before the experiment started they were tagged individually withpassive integrated transponders (PITs). Twelve individuals from each of six groups were mixed and released together at several occasions in the upper end of the 400-m-long experimental stream. An automatic PIT-monitoring system placed in the outlet recorded the time for passage of each individual leaving the stream. Eighty-five percent of the juveniles monitored by the PIT antenna showed downstream migration at night hours and they migrated significantly more often closer in time to both known and unknown siblings than to unrelated fish. The results suggest that there is a genetic component in the migratory behaviour of Atlantic salmon smolts and support the hypothesis that smolts migrate in kin-structured groups.

Laboratory studies in domesticated goldfish (Carassius auratus) show that, during the preovulatory luteinizing hormone (LH) surge, females release a complex steroidal pheromone that induces in males a rapid increase of plasma LH, which in turn increases strippable milt (sperm and seminal fluid) prior to ovulation and spawning. The objective of this study was to determine if the same phenomenon occurs in a wild congener, the crucian carp (Carassius carassius), under field conditions where fish are held in natural waters under ambient temperature and photoperiod. During the spawning season in June 2003, crucian carp were trapped in a small pond near Uppsala, Sweden, and held separately by sex in floating net pens. Addition of untreated females to male pens did not change male LH concentrations or milt volume during the 17 h sampling period. In contrast.. addition of females injected with Ovaprim (TM) (to induce an LH surge and ovulation) increased male LH concentrations at all sample times (5, 9, 13, and 17 h) following female addition and increased milt volumes at all but the first (5 h) sample time. Similar increases in male LH and milt that also occurred when untreated females ovulated spontaneously after addition to male pens suggest it is female ovulatory condition, rather than injection of ovaprim per se, that induced male LH and milt responses. Males also increased LH and milt 9 It after addition of females injected with the goldfish pheromonal steroid 4-pregnen-17,20 beta-diol-3-one (17,20 beta P), suggesting that similar responses to ovaprim-injected females were due, at least in part, to release of preovulatory pheromonal steroid(s). The clear and consistent effects of ovulatory females on male LH and milt, and the fact that crucian carp adapted well to confinement.. ovulated spontaneously, and exhibited apparently normal spawning behavior, all suggest that this species can serve as a useful cyprinid model to study reproductive processes in natural conditions.

In fish aquaculture, great efforts are made to develop feed that excludes marine fish proteins and oils, or at least to decrease these parts to a minimum but the fish might not be attracted to or might not eat the alternative feed. Molecules detected by the olfactory sense can induce arousal and search behavior that attract the fish and induce foraging. In the present study, we have examined whether mussel extracts can induce foraging behavior in crucian carp, Carassius carassius. Free amino acids are present in high concentrations in mussels and they are detected at low concentrations by olfaction and induce food search behavior in several fish species. We studied the behavior responses to water extracts of mussel powder compared to extracts of a plant-based commercial carp feed. Extracts of different concentrations were administered in glass aquaria with individual fish. The tests began with 5 min with only tap water and this was followed by 5 min with extracts or amino acid solutions. The behavior was recorded with a video camera. The initial control period was compared to the following period with odors. No significant foraging response was observed with extracts of mussels, but the commercial carp feed extract induced increased bottom snapping. Synthetic mixtures of five nutritional amino acids present at high concentrations in both extracts did not give any significant foraging response. Our results show that mussels are not suitable as a protein source in feed to crucian carp. 

The importance of learning for sibling odor preference in juvenile Arctic char was analyzed in the present study. Fish were reared in the following eight conditions: (1) communally with siblings for 15 months; (2) communally with siblings for 17 months; (3) in isolation since fertilization; (4) in isolation since fertilization and exposed to sibling scent during the whole rearing period; (5) in isolation since fertilization and exposed to sibling scent from time of free swimming; (6) in isolation since fertilization and exposed to sibling scent during the whole rearing period, except two months without scent until testing; (7) in isolation since fertilization and exposed to sibling scent from time of free swimming, except two months without scent until testing; and (8) communally with siblings followed by a two-month isolation until testing. Char were followed individually in a Y-maze (fluviarium test) with a video-computer-based image analysis system for 12 hr. Sibling-scented water was supplied to one lateral half of the test area and water from non-siblings on the opposite half. Isolated individuals without any preexposure to siblings showed no significant preference. Test fish reared with siblings and those that had been reared in isolation but exposed to sibling scent until testing preferred water conditioned by their own siblings. Isolated fish that had been exposed to sibling scent since fertilization, or since free swimming, followed by a two-month period with only pure water, showed no significant preference. Char isolated for two months after being communally reared preferred water scented by siblings. The results demonstrated that behavioral discrimination between siblings and nonsibling odors occurred after total isolation (isolated both from siblings and sibling odors) only in individuals that had been communally reared. This may suggest that social interactions are important for learning and long-term memory of sibling odors in Arctic char.

Several studies have shown that fish shoals may consist of closely related individuals. It has been found. for example, that released out-migrating salmon smolts tend to aggregate with kin. including when sibling groups have been reared separately. We used genetic microsatellite markers to test whether "shoals" of adult Atlantic salmon (Salmo salar) during the marine phase (i.e., aggregations of fish Caught in drift nets at offshore feeding areas in the Baltic Sea) consisted of closely related individuals (full-siblings, half-siblings). We found no evidence of kin cohesiveness related to shoals, however. Despite a weak overall tendency for individuals assigned to the same population (river or stock) to Occur tooether, estimates of genetic relatedness in combination with consistent heterozygote deficiencies. and results from mixed-stock analyses and assignment tests collectively indicated that shoals consisted of unrelated fish from multiple populations.