Developmental exposure to the SSRI citalopram causes long-lasting behavioural effects in the three-spined stickleback (Gasterosteus aculeatus).

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 long-lasting behavioural effects of pre- and perinatal exposure to SSRIs which last into adulthood. In fish however, studies on effects of developmental exposure to SSRIs appears to be non-existent. In order to study effects of developmental SSRI 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 approximately 100 days of remediation in clean water the fish were put through an extensive battery of behavioural tests. 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 min 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 long-lasting behavioural effects in fish and contribute to the existing knowledge about SSRIs as environmental pollutants.

Waterborne citalopram has anxiolytic effects and increases locomotor activity in the three-spine stickleback (Gasterosteus aculeatus).

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.

Environmentally relevant concentrations of citalopram partially inhibit feeding in the three-spine stickleback (Gasterosteus aculeatus).

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.

Candidate epitopes for measurement of hCG and related molecules: the second ISOBM TD-7 workshop.

Participants of the Second International Workshop (WS) on human chorionic gonadotropin (hCG) of the International Society of Oncology and Biomarkers Tissue Differentiation 7 (ISOBM TD-7) have characterized in detail a panel of 69 antibodies (Abs) directed against hCG and hCG-related variants that were submitted by eight companies and research groups. Specificities of the Abs were determined using the First WHO International Reference Reagents for six hCG variants, i.e., hCG, hCGn, hCGß, hCGßn, hCGßcf, and hCGα, which are calibrated in SI units, and hLH. Molecular epitope localizations were assigned to the ISOBM-mAbs by comparing ISOBM-Ab specificity, sandwich compatibility, and mutual inhibition profiles, to those of 17 reference monoclonal (m)Abs of known molecular epitope specificities. It appeared that 48 Abs recognized hCGß-, 8 hCGα-, and 13 αß-heterodimer-specific epitopes. Twenty-seven mAbs were of pan hCG specificity, two thereof with no (<0.1%; epitope ß1), 12 with low (<1.0%; epitopes ß2/4), and 13 with high (>>1%; epitopes ß3/5) hLH cross-reactivity. The majority of hCGß epitopes recognized were located in two major antigenic domains, one on the peptide chain of the tips of ß-sheet loops 1 and 3 (epitopes ß2-6; 27 mAbs) and the second around the cystine knot (e.g., epitopes ß1, ß7, and ß10; 9 mAbs). Four mAbs recognized epitopes on hCGßcf-only (e.g., epitopes ß11 and ß13) and six mAbs epitopes on the remote hCGß-carboxyl-terminal peptide (epitopes ß8 and ß9 corresponding to amino acids 135-144 and 111-116, respectively). For routine diagnostic measurements, methods are used that either detect hCG-only, hCGß-only, or hCG together with hCGß or hCG together with hCGß and hCGßcf. Sandwich assays that measure hCG plus hCGß and eventually hCGßcf should recognize the protein backbone of the analytes preferably on an equimolar basis, should not cross-react with hLH and not be susceptible to blunting of signal by nonmeasured variants like hCGßcf. Such assays can be constructed using pairs of mAbs directed against the cystine knot-associated epitope ß1 (Asp10, Asp60, and Gln89) in combination with epitopes ß2 or ß4 located at the top of ß-sheet loops 1 + 3 of hCGß involving aa hCGß20-25 + 68-77. In summary, the results of the First and Second ISOBM TD-7 WSs on hCG provide the basis for harmonization of specificities and epitopes of mAbs to be used in multifunctional and selective diagnostic hCG methods for different clinical purposes.

Molecular identification of hybrids of the invasive gibel carp Carassius auratus gibelio and crucian carp Carassius carassius in Swedish waters.

Both mitochondrial DNA sequence and two nuclear microsatellite markers were used to confirm the identity of the first record of Carassius auratus gibelio in the western (Swedish) Baltic Sea region. A total of 49 fishes were analysed, where 22 were from three Swedish sites connected to the Baltic Sea. The D-loop mitochondrial DNA sequences showed that 16 of 22 Swedish fishes were related to C. a. gibelio. The phylogenetic analysis of these sequences showed that these fish are probably not native, but represent different lineages of C. a. gibelio from China, Japan and Russia. All except three of these 16 fishes had microsatellite alleles suggesting hybridization with Carassius carassius. These findings suggest that a cryptic invasion of C. a. gibelio might be in progress.

Male primer endocrine responses to preovulatory female cyprinids under natural conditions in Sweden.

This study investigated two related aspects of male-female reproductive interactions in the family Cyprinidae: (1) whether ovulating female rudd Scardinius erythrophthalmus (subfamily Leuciscinae) induce endocrine and gonadal priming responses in conspecific males, a phenomenon which has been described only in species from the subfamily Cyprininae such as goldfish, Carassius auratus, crucian carp Carassius carassius and common carp, Cyprinus carpio and (2) whether the stimuli mediating these responses are species-specific. Field studies of three sympatric European cyprinids, two leuciscins (S. erythrophthalmus and white bream Blicca bjoerkna) and one cyprinin (C. carassius), were conducted on fishes captured in Sweden in the spawning season and held in net pens under natural conditions. As previously reported in C. carassius, male S. erythrophthalmus increased milt (sperm and seminal fluid) volume and plasma concentrations of the sperm maturation hormone 4-pregnen-17,20ß-diol-3-one (17,20ß-P) when they were held with female S. erythrophthalmus induced to ovulate by injection of Ovaprim (GnRH analogue plus dopamine antagonist). Male S. erythrophthalmus had larger milt volumes than male C. carassius prior to and following exposure to ovulatory conspecifics, but exhibited a smaller proportional milt increase in response to stimulation, suggesting species differences in sperm allocation at spawning. The presence of female S. erythrophthalmus and B. bjoerkna did not affect milt volumes of C. carassius under two experimental conditions: (1) ovulating S. erythrophthalmus and B. bjoerkna did not increase the milt volumes of C. carassius and (2) S. erythrophthalmus and B. bjoerkna did not interfere with the milt volume increase induced in male C. carassius by ovulating conspecifics. These results suggest that, as in C. auratus, C. carassius and C. carpio (subfamily Cyprininae), female S. erythrophthalmus (subfamily Leuciscinae) release a preovulatory pheromone that exerts priming effects on male hormones and sperm allocation. The findings also indicate that C. carassius discriminate between the reproductive odours of conspecifics and heterospecifics.

Endocrine and milt responses of male crucian carp (Carassius carassius L.) to periovulatory females under field conditions.

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 (to induce an LH surge and ovulation) increased male LH concentrations at all sample times (5, 9, 13, and 17h) following female addition and increased milt volumes at all but the first (5h) 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 9h after addition of females injected with the goldfish pheromonal steroid 4-pregnen-17,20beta-diol-3-one (17,20betaP), 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.

Kin-biased distribution in brown trout: an effect of redd location or kin recognition?

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.

Chemobehavioral changes induced by short-term exposures to prochloraz, nicosulfuron, and carbofuran in goldfish.

The behavioral effects of short periods (2, 4, 6, 8 h) of static exposure to prochloraz (imidazole fungicide) and nicosulfuron (sulfonylurea herbicide) were recorded in goldfish (Carassius auratus). Observations were also made in an olfactometer to assess the effects of 8-h exposures to these two pesticides and to carbofuran (carbamate insecticide) on the behavioral responses to the flow of a solution of four L-amino acids (glycine, alanine, valine, taurine), mixed in the same relative proportions as in the urine of conspecifics. Each pesticide was tested at three sublethal concentrations (25, 50, 100 microg/L), and the behaviors recorded were related to swimming pattern, social interactions, and comfort movements. Static exposures to prochloraz affected horizontal displacements, burst swimming, grouping, and buccal movements. Static exposures to nicosulfuron affected burst swimming and grouping. In pesticide-unexposed fish (control), the flow of the amino acid solution induced attraction, decreased sheltering, and increased horizontal displacements, burst swimming, buccal movements, and antagonistic interactions. Compared to the controls, some of the behavioral responses to the solution of amino acids were significantly different after 8 h of subacute exposure to prochloraz and carbofuran. Both pesticides decreased attraction and increased sheltering. In addition, carbofuran decreased buccal movements and antagonistic interactions. Contrastingly, exposure to nicosulfuron showed no significant effect. This study further confirms the great vulnerability of fish behavior and chemocommunication processes to exposure to waterborne pesticides.

Influence of MHC on sibling discrimination in Arctic char, Salvelinus alpinus (L.).

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.

Epitopes on CA 125 from cervical mucus and ascites fluid and characterization of six new antibodies. Third report from the ISOBM TD-1 workshop.

CA 125 is found in body fluids in a variety of molecular weight forms. The largest species are found in normal abdominal fluid and cervical mucus. The present study therefore incorporated CA 125 derived from these sources as well as ascites fluid to investigate if the source of CA 125 influenced epitope characterization. Ascites-derived CA 125 varied in size from about 190 to about 2,700 kD. Cervical mucus-derived CA 125 treated with ultrasound changed its apparent size from more than 20,000 to 700 kD. Epitope mapping of antibodies was not grossly influenced by the size or source of CA 125 used as target. However, low-molecular-weight CA 125, i.e. ascites fractions CA 17/E, CA 17/F and CA 10/7, did show differences in certain assay combinations and cross-inhibition patterns which probably can be explained by steric effects due to the smaller size compared with the most abundant forms of CA 125 present in serum and other body fluids. The specificity of six new monoclonal antibodies to CA 125 was tested by cross-inhibition and immunometric assay combinations and compared to reference antibodies. One antibody, X306, belonged to the OC125-like antibodies. Four antibodies, X52, X75, X325 and VK8, were M11-like. The sixth antibody, 7C12, reacted with an epitope which was difficult to define. This antibody was inhibited by M11-like antibodies and OV197. However, used as an inhibitor, 7C12 inhibited only itself. We grouped it as an OV197-like antibody, but clearly different from OV197. The topography of epitopes was studied by analyzing all antibody pairs in immunoradiometric assays. These results confirmed the grouping of antibodies described above and are in accordance with previous findings that the highest signal is obtained using an OC125-like antibody or OV197 on the solid phase and an M11-like antibody as tracer. The composition of the sample in terms of high- and low-molecular-weight species of CA 125 was measured, with different responses depending on the antibody pair used. This might be one reason for discrepancies between assay results for CA 125 using different assays.

Transoesophageal echocardiographic monitoring during paediatric cardiac surgery: obtainable information and feasibility in 532 children.

BACKGROUND: We hypothesized that transoesophageal echocardiography (TOE) performed by the anaesthesiologists would be beneficial for monitoring purposes during paediatric cardiac surgery. We present the results for the first 5 years in 532 consecutive children. METHODS: The probe was successfully inserted in 99% of cases and remained in the oesophagus for 211 min on average (range 10-555 min). RESULTS: Insignificant valve leak, single- or biventricular failure and volume depletion were the most common new findings due to TOE. Changes in inotropic strategy and volume replacement were the most frequent interventions. In 45% of the cases, new information was disclosed and, in a total of 8% of cases, decisive information was provided. Except for tracheal extubation in one child who was uneventfully reintubated, no severe complications were identified. CONCLUSIONS: These data stress the safety and ease of performing TOE in children undergoing cardiac surgery. There is evidence for benefit from TOE findings to potentially enhance the therapeutic basis.

Behavioral and olfactory responses to prochloraz, bentazone, and nicosulfuron-contaminated flows in goldfish.

The immediate behavioral responses of goldfish (Carassius auratus) to pesticide-contaminated flows were recorded in a countercurrent olfactometer. In addition, electro-olfactograms were recorded from the epithelial surface of the olfactory rosette as a preliminary check for the olfactory sensitivity of the fish to the pesticides tested. All tests were run on prochloraz (imidazole fungicide), bentazone (diazine herbicide), and nicosulfuron (sulfonylurea herbicide). Behavioral effects were assessed, at four concentrations (10 microg/L, 100 microg/L, 1 mg/L, 10 mg/L), on endpoints related to swimming pattern (preference-avoidance responses, burst swimming reactions), comfort activities (buccal movements, feeding attempts), and social relations (antagonistic acts, grouping). The behavior of the fish appeared particularly sensitive to prochloraz exposure. As a whole, prochloraz-contaminated flows showed significant effects on the six behaviors studied; bentazone and nicosulfuron affected three and five, respectively. At the lowest concentration, prochloraz also showed more effects than the two other pesticides. Some of the behavioral endpoints were found particularly sensitive to a given chemical. Pesticide-contaminated flows also induced significant changes in swimming orientation of the fish. Attraction was observed in response to flowing solutions of prochloraz (1 mg/L, 10 mg/L), bentazone (10 microg/L, 10 mg/L), and nicosulfuron (1 mg/L, 10 mg/L). At a concentration of 1 mg/L, none of the pesticides induced a noticeable depolarization of the olfactory epithelium, suggesting that these chemicals are not detected by the olfactory sense of the fish. These results are discussed in the light of the data concerning effects of pesticides on behavior and chemical communication in fish.

Both ovarian fluid and female urine increase sex steroid hormone levels in mature Atlantic salmon (Salmo salar) male parr.

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 F2alpha (PGF2alpha) 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 PGF2alpha. Plasma concentrations of the sex steroids of 17,20beta-dihydroxy-4-pregnen-3-one (17,20beta-P) were higher in males exposed to urine, ovarian fluids, and PGF2alpha compared to control males. PGF2alpha and a mixture of urine and ovarian fluid also gave increased concentrations of 11-ketotestosterone (11-KT). Concentrations of PGF2alpha were higher in ovarian fluids than in urine. A behavior test with mature male parr in a fluviarium showed neither attraction to nor avoidance of 0.1 nM PGF2alpha, but plasma levels of 17,20beta-P were significantly higher in exposed males compared to controls.

Neurohormonal activation late after cavopulmonary connection.

OBJECTIVE: To determine whether patients with cavopulmonary connection have higher levels of vasoactive/water-salt regulating hormones and if so, whether hormone levels are related to postoperative haemodynamics and postoperative follow up. DESIGN: Cross sectional study. SETTING: University hospital. PATIENTS: 20 patients (New York Heart Association functional class I-II), mean age 11 years (range 4 to 22), were studied at a mean of 2 years (0.5 to 6) after a total cavopulmonary connection (TCPC, n = 12) or a bidirectional Glenn anastomosis (BDG, n = 8). INTERVENTIONS: Cardiac catheterisation was performed and blood samples were drawn. Control blood samples were drawn from 33 healthy children, mean age 12 years (6 to 16). MAIN OUTCOME MEASURES: Plasma levels of angiotensin II, renin, aldosterone, arginine, vasopressin, atrial natriuretic factor (ANF), brain natriuretic peptide (BNP). RESULTS: All neurohormones were significantly increased in both TCPC and BDG patients (p < 0. 05), with a fourfold increase in angiotensin II, renin, and aldosterone, and a twofold increase in vasopressin, ANF, and BNP (compared with healthy controls). There was no correlation between haemodynamic variables and hormone levels. Angiotensin II and renin were inversely correlated with time to follow up. All subjects over 15 years (n = 5) had normal neurohormonal levels. CONCLUSIONS: Neurohormones were raised for years after successful cavopulmonary operations but lower levels were observed with time on follow up. This supports the hypothesis that neurohormonal activation is primarily related to altered postoperative physiology and that adaptation takes place over time.

Pediatric multiplane transesophageal echocardiography in congenital heart disease: new possibilities with a miniaturized probe.

In recent years, transducers for multiplane Doppler echocardiography have demonstrated their superior imaging performance in adult patients. To date, the size of these probes has limited their use in pediatric patients. In this article, we report our initial experience with a recently developed miniaturized transducer with all conventional imaging and Doppler modalities. The study focused primarily on imaging performance by comparing standard biplane images with those obtained in oblique planes. The investigations were carried out intraoperatively or during interventional catheterization in patients with congenital heart disease. We observed no complications in a study population of 15 children (weight range of 5 to 63 kg and an age range of 96 days to 11 years). The probe was easy to handle and provided excellent images. Additional information was obtained in several cases and documentation of clinical findings was easier because an optimal image plane almost always could be displayed. We concluded that pediatric multiplane Doppler echocardiography has considerably improved investigative performance compared with the conventional monoplane or biplane studies normally available for this age group. In neonates, however, investigation with the multiplane technique is limited by the size of the patient.

Endocrine, gonadal and behavioral responses of male crucian carp to the hormonal pheromone 17 alpha,20 beta-dihydroxy-4-pregnen-3-one.

The olfactory-mediated responses to the sex hormone 17 alpha,20 beta-dihydroxy-4-pregnen-3-one (17 alpha,20 beta-P) were studied in spermiated and regressed male crucian carp (Carassius carassius L.). The position and spontaneous locomotor activity of single male crucian carp were continuously recorded in an artificial stream. 17 alpha,20 beta-P (final concentration 10(-11) M) was supplied to one half and its ethanol carrier to the other half of the test area. Milt volume and gonadotropin (GtH-II) concentration in the plasma were also measured. The smell of 17 alpha,20 beta-P significantly increased both the GtH-II concentration in the plasma and the volume of strippable milt in spermiated crucian carp. Behaviorally, the side of the test area scented with 17 alpha,20 beta-P was significantly avoided by spermiated males. None of the described effects of 17 alpha,20 beta-P on spermiated males were observed for the regressed crucian carp. In view of the lack of response from regressed crucian carp we suggest that the observed avoidance behavior of 17 alpha,20 beta-P by spermiated males is a relevant reaction for spawning male crucian carp. The results are well in accordance with responses obtained in the closely related goldfish and gives strong support that the wild male crucian carp use the 17 alpha,20 beta-P signal from the females to prepare for the coming spawning.

Endocrine changes associated with spawning behavior in male kokanee salmon (Oncorhynchus nerka) and the effects of anosmia.

This study examines the relationship between social status, behavioral activity, and hormone levels through the spawning cycle in male kokanee salmon (Oncorhynchus nerka). Shortly after arrival at the spawning grounds, male kokanee undergo dramatic changes in morphological and physiological condition: there is an increase in body depth, a bright silvery appearance is replaced by an intense red coloration, and spermiation commences. Males become associated with nesting females which they defend against other males. The androgens, testosterone and 11-ketotestosterone, decline slowly over the spawning period from high levels observed in the silvery males; gonadotropin (GtH II) and 17 alpha, 20 beta-dihydroxy-4-pregnen-3-one increase as males approach final maturation. Androgen and GtH levels in males placed in pens with females over the spawning bed were higher than those in males penned without females. Among males competing for females, amounts of milt and plasma concentrations of GtH were higher in those males that were successful in gaining access to nesting females. A marked reduction in milt and plasma hormones following cauterization of the olfactory nares indicates that olfactory stimuli play a role in mediating endocrine and spermiation responses to the nesting females. Anosmic males were less vigorous and persistent in their courtship, suggesting that, either a female pheromone and/or high levels of hormone are necessary to maintain full reproductive behavior in male kokanee. The role of socially induced hormone changes in modulating reproductive behavior is discussed.

The significance of olfaction and social cues in milt availability, sexual hormone status, and spawning behavior of male rainbow trout (Oncorhynchus mykiss).

Male rainbow trout orientate to a "releaser" pheromone emitted by an ovulated female. This chemical signal is not necessary for the completion of spawning behavior: anosmic males spawned as readily as intact males when paired with ovulated females. The amount of "strippable" milt and plasma concentrations of testosterone (T) and 17 alpha,20 beta-dihydroxy-4-pregnen-3-one (17,20 beta-P) were greater in males paired for 3 hr with ovulated females than in isolated males, or males placed with nonvitellogenic adult females or in all male groups. Milt and plasma T, 17,20 beta-P, and 11-ketotestosterone (11-KT) were reduced in anosmic males paired with ovulated females, indicating the existence of a priming pheromone. Milt, T, 11-KT, and 17,20 beta-P increased in intact males within 4 hr after injection of salmon gonadotrophin-releasing hormone analogue, lyophilized salmon pituitary extract, or homogenized fresh rainbow trout pituitary material. The results indicate that the priming pheromone acts through the hypothalamus-pituitary-gonad axis.

Changes in brain serotonergic activity during hierarchic behavior in Arctic charr (Salvelinus alpinus L.) are socially induced.

The experiment was performed in two phases. During the first phase (phase 1) the dominance hierarchy was determined in 4 groups of Arctic charr (Salvelinus alpinus L.), each group consisting of 4 fish. Phase 2 was started by rearranging phase 1 fish into 4 new groups. Group 1 consisted of previously dominant fish and groups 2, 3 and 4 of fish that previously held rank 2, 3 and 4, respectively. After phase 2 telencephalon and brain stem were analyzed with regard to their contents of serotonin (5-hydroxytryptamine, 5-HT) and 5-hydroxyindoleacetic acid (5-HIAA), the principle metabolite of 5-HT. No correlation was found between the social rank (measured as dominance index) during phase 1 and the brain serotonergic activity (measured as the ratio 5-HIAA/5-HT) determined after phase 2. However, most important, the 5-HIAA/5-HT ratio was significantly correlated with the last experienced social rank, i.e. that acquired during phase 2. These results shows that the difference in brain serotonergic activity between dominant and subordinate fish develops through social interactions. Further, we found that previous subordinate experience inhibited aggressive behavior, an effect which, in the light of available information on stress and 5-HT, could be related to the increase in brain serotonergic activity. We hypothesize that stress induces an increased serotonergic activity which in turn inhibits the neuronal circuitry which mediates aggressive behavior.