Male Three-spined Sticklebacks Gasterosteus Aculeatus Research Articles

New finding of melanic three-spined sticklebacks Gasterosteus aculeatus in the Scottish Hebrides

Colour traits can be elaborated through sexual selection and have potential to drive reproductive isolation. Male three-spined sticklebacks (Gasterosteus aculeatus) express striking visual signals to attract choosy females during courtship, typically expressed as red carotenoid-based pigmentation on their throat and jaw during the breeding season, along with blue eyes and blue/green flanks. The extent and intensity of red colouration in males have been linked to fitness benefits to females, including body condition, parasite resistance, parental ability and nest defence. In some populations in the Pacific Northwest of North America, male three-spined sticklebacks express melanic nuptial colouration. In these populations, male possess black throats instead of red, and have dark or black bodies. Melanic males are associated with waterbodies that are red-shifted due to the presence of tannins, where the ambient light environment is dominated by long wavelengths. Here we report the first discovery outside North America of melanic populations of three-spined sticklebacks on the island of North Uist in the Scottish Hebrides, on the northwest Atlantic coast of Europe. These populations are associated with a hotspot of stickleback morphological diversity and occur in association with red-shifted waterbodies.

Differential investment in pre- and post-mating male sexual traits in response to prolonged exposure to ambient UVB radiation in a fish

Increasing UVB radiation (UVB) reaching earth's surface following stratospheric ozone depletion is linked to serious consequences for organisms. While studies have focused on direct cytocidal and immunomodulatory effects of UVB, indirect consequences for fitness-related life-history traits are largely unexplored, although knowledge is needed to understand organismal responses to climate change. The present study investigates the effects of developmental exposure to enhanced, but ecologically relevant, ambient UVB levels on growth (body size), parental behavior (nest-building), fitness-enhancing traits of pre-mating (sexual ornamentation) and post-mating (sperm traits) sexual selection as well as their interrelationships in male three-spined sticklebacks (Gasterosteus aculeatus). Moreover, potentially underlying direct UVB effects, testicular antioxidant capacity (TAC) and testes and skin melanization were quantified. Juvenile full-siblings were split into two groups and raised until adulthood in a semi-natural set-up under light conditions including either natural (UVBnormal) or elevated UVB levels (UVBenhanced). When becoming reproductive, males were kept individually before data collection took place at their reproductive peak. The results showed that males from the UVBenhanced-group were smaller than their brothers from the UVBnormal-treatment, whereas no treatment differences were observed for sexual ornamentation, sperm traits and TAC. Moreover, UVB-stressed males built their nests faster and the relationship between body size and nest size was negative for UVBenhanced-males while being positive for UVBnormal-males. These results demonstrate that, depending on physical state, UVB-stressed males adjusted their behavior to some extent. Additionally, a trade-off between the investment in pre- and post-mating traits was found for UVBenhanced-males, i.e. the intensity of their breeding coloration was negatively correlated with sperm number whereas this relationship was reversed for UVBnormal-males, thus showing an interaction between pre- and post-mating traits regarding exposure treatment. The interaction provides first experimental evidence that differential allocation to energetically demanding pre- and post-mating components of male fitness is triggered by a key environmental stressor of climate change.

Natural variation in brain gene expression profiles of aggressive and nonaggressive individual sticklebacks.

Within many species, some individuals are consistently more aggressive than others. We examine whether there are differences in brain gene expression between aggressive versus nonaggressive behavioural types of individuals within a natural population of male three-spined sticklebacks (Gasterosteus aculeatus). We compared gene expression profiles of aggressive male sticklebacks to nonaggressive males in four regions of the brain (brainstem, cerebellum, diencephalon and telencephalon). Relatively few genes were differentially expressed between behavioural types in telencephalon, cerebellum and diencephalon, but hundreds of genes were differentially expressed in brainstem, a brain area involved in detecting threats. Six genes that were differentially expressed in response to a territorial intrusion in a previous study were also differentially expressed between behavioural types in this study, implying primarily non-shared but some shared molecular mechanisms. Our findings offer new insights into the molecular causes and correlates of behavioural plasticity and individual variation in behaviour.

Prozac affects stickleback nest quality without altering androgen, spiggin or aggression levels during a 21-day breeding test

Pharmaceuticals are increasingly being used in human and veterinary medicine, and their presence in the aquatic environment may present a threat to non-target aquatic organisms. The selective serotonin reuptake inhibitor fluoxetine (Prozac) has been reported to affect diverse behaviours (feeding, aggression, and reproduction) and also the endocrine system (steroid biosynthesis pathway) in fish. To investigate these claims further, and in particular effects on androgen synthesis, male three-spined sticklebacks (Gasterosteus aculeatus) were exposed to fluoxetine at 0, 3.2, 10 and 32μg/L in a flow-through system for 21 days. Their sex was determined prior to exposure using a non-invasive method to collect DNA for determining the genetic sex, reported here for the first time. This was necessary as the exposure required males of a non-breeding status which had not developed secondary characteristics. Post exposure a number of biochemical (serotonin, steroid and spiggin levels) and apical (aggressive behaviour) endpoints were measured. No effects were detected on morphometric parameters, spiggin or androgen (11-ketotestosterone) levels. However, all fluoxetine-exposed male fish had higher cortisol levels in comparison to the control fish, although this effect only persisted throughout the whole exposure duration at the highest concentration (32μg/L). In addition, the ratio of 5-HIAA/5-HT (serotonin metabolite/serotonin) was significantly lower in the brains of males exposed to fluoxetine at all concentrations tested. Although we found no differences in the number of nests built by the males, the quality of the nests produced by the fluoxetine-exposed males was generally inferior consisting only of a basic, rudimentary structure. Males exposed to 32μg/L of fluoxetine displayed a delayed response to a simulated threat (rival male via own mirror image) and were less aggressive (number of bites and attacks) toward their mirror image, but these differences were not statistically significant. In summary, fluoxetine exposure resulted in reduced serotonergic activity in the male three-spined stickleback brain suggesting that the mechanism of action between humans and fish is at least partially conserved. Furthermore, this study provided additional evidence of cross-talk between the serotonergic and stress axes as demonstrated by the perturbations in cortisol levels. This potentially complex interaction at brain level may be responsible for the effects observed on nest quality, an endpoint with serious ecological consequences for this species. Finally, despite our hypothesis (an effect on steroid biosynthesis, based on limited literature evidence), we observed no effects of fluoxetine exposure (at the concentrations and duration employed) on male stickleback androgen levels.

Reproductive strategies and seasonal changes in the somatic indices of seven small-bodied fishes in Atlantic Canada in relation to study design for environmental effects monitoring.

Small-bodied fishes are more commonly being used in environmental effects monitoring (EEM) studies. There is a lack of understanding of the biological characteristics of many small-bodied species, which hinders study designs for monitoring studies. For example, 72% of fish population surveys in Canada's EEM program for pulp and paper mills that used small-bodied fishes were conducted outside of the reproductive period of the species. This resulted in an inadequate assessment of the EEM program's primary effect endpoint (reproduction) for these studies. The present study examined seasonal changes in liver size, gonad size, and condition in seven freshwater and estuarine small-bodied fishes in Atlantic Canada. These data were used to examine differences in reproductive strategies and patterns of energy storage among species. Female gonadal recrudescence in all seven species began primarily in the 2-month period in the spring before spawning. Male gonadal development was concurrent with females in five species; however, gonadal recrudescence began in the fall in male three-spined stickleback (Gasterosteus aculeatus) and slimy sculpin (Cottus cognatus). The spawning period for each species was estimated from the decline in relative ovary size after its seasonal maximum value in spring. The duration of the spawning period reflected the reproductive strategy (single vs multiple spawning) of the species. Optimal sampling periods to assess reproductive impacts in each species were determined based on seasonal changes in ovary size and were identified to be during the prespawning period when gonads are developing and variability in relative gonad size is at a minimum.

Male Mating Posture and Courtship Coloration of the Temperate Marine Tubesnout,Aulorhynchus flavidus(Gill)

Species that engage in sexual reproduction often exhibit dynamic physical features and behavioral actions that serve to signal conspecifics. Body posture, vocalizations or the incorporation of bright coloration are typically used to establish dominance (Wiley 1973, Warner 1975, Clarke and Faulkes 1997, Peterson and Jacobs 2002) alert conspecifics of nearby threats or foraging opportunities (Godin and Morgan 1985, Beauchamp and Heeb 2001, Le Roux et al. 2009, Townsend et al. 2011) and for breeding purposes (Wiley 1973, Hagen et al. 1980, Christy 1983, Gibson 1989, DeMartini and Sikkel 2006). In many species females select a male based on their behavioral display of some desirable trait or their ability to express dominance or competitive superiority over other potential mates (DeMartini and Sikkel 2006). Such criteria are often not mutually exclusive in that particular features or actions may act to deter competitor males while simultaneously attracting females (Hagen et al. 1980, Cairns 1982). Understanding such physical characteristics or behavioral signals improves our knowledge of reproductive systems within and among taxonomic groups. Reproductive strategies have been well documented in the marine realm, including monogamous pairs (Whiteman and Cote 2004), large broadcast breeding aggregations (Coleman et al. 1996, Colin 1996, Carolsfeld et al. 1997), sneak copulation (Pilastro and Bisazza 1999, Hurtado-Gonzales and Uy 2010) and nesting species (Limbaugh 1964, Wiley 1973). Limbaugh (1962) previously described the nesting habitat for the temperate marine tubesnout, Aulorhynchus flavidus (Gill), but male mating behavior has not been characterized. A proposed description of male mating behavior is reported here from observations of A. flavidus on display at the Aquarium of the Pacific (AOP) in Long Beach, California during early May, 2013. Male A. flavidus pelvic fins are red in color (Fig. 1A) and are thought to remain that way year round (Love 2011). The heads of males are also dark blue to black with a prominent iridescent blue patch on their snout (Fig. 1B). Iridescent blue spots/coloration have also been observed along the full body length of males (Love 2011) while females are a drab brownish color throughout (Fig. 2). Although typically found at depths of 20 m, and as deep as 37 m (Love 2011), the preferred nesting habitat of A. flavidus is the upper blades of the macrophyte Macrocystis pyrifera. While bright coloration is common to tropical waters, it is disfavored in temperate marine habitats due to low light penetration at depth. The conspicuous coloration of males may be apparent only when actively breeding in the upper water column where red light is still present. It appears the pelvic fins are often folded up against the body (Fig. 1B) and covered partially by the pectoral fins when they are not actively displaying them. The anterior underside of male threespine stickleback (Gasterosteus aculeatus), another gasteriform species, are also red (Hagen et al. 1980) and both species breed in shallower water than they are typically found (,20– 30 m depth). Similarities in external morphology despite differences in breeding habits Bull. Southern California Acad. Sci. 113(3), 2014, pp. 176–179 E Southern California Academy of Sciences, 2014

GnRH mRNA levels in male three-spined sticklebacks, Gasterosteus aculeatus, under different reproductive conditions

In vertebrates, reproduction is regulated by the brain–pituitary–gonad (BPG) axis, where the gonadotropin-releasing hormone (GnRH) is one of the key components. However, very little is known about the possible role of GnRH in the environmental and feedback control of fish reproduction. To investigate this, full-length gnrh2 (chicken GnRH II) and gnrh3 (salmon GnRH) sequences of male three-spined sticklebacks (Gasterosteus aculeatus), which are clustered with the taxa of the same GnRH type as other Euteleostei, were cloned and annotated. gnrh1 is absent in this species. The mRNA levels of gnrh2 and gnrh3 in the sticklebacks' brain were measured under breeding and post-breeding conditions as well as in castrated and sham-operated breeding fish and castrated/sham-operated fish kept under long-day (LD 16:8) and short-day (LD 8:16) conditions. Fully breeding males had considerably higher mRNA levels of gnrh2 and gnrh3 in the thalamus (Th) and in the telencephalon and preoptic area (T+POA), respectively, than post-breeding males. Sham-operated breeding males have higher gnrh3 mRNA levels than the corresponding castrated males. Moreover, higher gnrh2 mRNA levels in the Th and higher gnrh3 mRNA levels in the T+POA and hypothalamus (HypTh) were also found in long-day sham-operated males than in sham-operated fish kept under an inhibitory short day photoperiod. Nevertheless, gnrh2 and gnrh3 mRNA levels were not up-regulated in castrated males kept under long-day photoperiod, which suggests that positive feedbacks on the brain-pituitary-gonad axis are necessary for this response.

Timescale effects of 17α-ethinylestradiol on behavioral consistency in male threespine stickleback

While the effects of 17α-ethinylestradiol (EE2) on male-typical behaviors are well studied, much less is known about the timing of these effects or the context in which these effects occur as most studies assay behavior at a single time point or in a single context. Additionally, exposure may affect behavior at levels beyond overall responsiveness, such as by altering behavioral consistency over time. Male threespine stickleback (Gasterosteus aculeatus) experienced three different contexts (single female, single male, paired female-male) over the course of 8 days, with an experimental group of males receiving EE2 after the first day. Exposure to EE2 disrupted responsiveness, behavioral consistency, and behavioral variation in all three social contexts. The timescale over which alterations in behavior occurred differed based on whether the conspecific was presented singly or paired with an opposite sex conspecific, stressing the importance of examining the effects of contaminants in multiple contexts.

Flow-mediated plasticity in the expression of stickleback nesting glue genes.

Nest construction is an essential component of the reproductive behavior of many species, and attributes of nests – including their location and structure – have implications for both their functional capacity as incubators for developing offspring, and their attractiveness to potential mates. To maximize reproductive success, nests must therefore be suited to local environmental conditions. Male three-spined sticklebacks (Gasterosteus aculeatus) build nests from collected materials and use an endogenous, glue-like multimeric protein – “spiggin” – as an adhesive. Spiggin is encoded by a multigene family, and differential expression of spiggin genes potentially allows plasticity in nest construction in response to variable environments. Here, we show that the expression of spiggin genes is affected significantly by both the flow regime experienced by a fish and its nesting status. Further, we show the effects of flow on expression patterns are gene-specific. Nest-building fish exhibited consistently higher expression levels of the three genes under investigation (Spg-a,Spg-1, and Spg-2) than non-nesting controls, irrespective of rearing flow treatment. Fish reared under flowing-water conditions showed significantly increased levels of spiggin gene expression compared to those reared in still water, but this effect was far stronger for Spg-a than for Spg-1 or Spg-2. The strong effect of flowing water on Spg-a expression, even among non-nesters, suggests that the increased production of spiggin – or of spiggin rich in the component contributed by Spg-a – may allow more rapid and/or effective nest construction under challenging high flow conditions.

Sex-Differences and Temporal Consistency in Stickleback Fish Boldness

Behavioural traits that co-vary across contexts or situations often reflect fundamental trade-offs which individuals experience in different contexts (e.g. fitness trade-offs between exploration and predation risk). Since males tend to experience greater variance in reproductive success than females, there may be considerable fitness benefits associated with “bolder” behavioural types, but only recently have researchers begun to consider sex-specific and life-history strategies associated with these. Here we test the hypothesis that male three-spined sticklebacks (Gasterosteus aculeatus) show high risk but potentially high return behaviours compared to females. According to this hypothesis we predicted that male fish would show greater exploration of their environment in a foraging context, and be caught sooner by an experimenter than females. We found that the time fish spent out of cover exploring their environment was correlated over two days, and males spent significantly more time out of cover than females. Also, the order in which fish were net-caught from their holding aquarium by an experimenter prior to experiments was negatively correlated with the time spent out of cover during tests, and males tended to be caught sooner than females. Moreover, we found a positive correlation between the catch number prior to our experiments and nine months after, pointing towards consistent, long-term individual differences in behaviour.

The Synthetic Progestin Levonorgestrel Is a Potent Androgen in the Three-Spined Stickleback (Gasterosteus aculeatus)

The use of progestins has resulted in contamination of aquatic environments and some progestins have in experimental studies been shown to impair reproduction in fish and amphibians at low ng L(-1) concentrations. The mechanisms underlying their reproductive toxicity are largely unknown. Some progestins, such as levonorgestrel (LNG), exert androgenic effects in mammals by activating the androgen receptor (AR). Male three-spined stickleback (Gasterosteus aculeatus) kidneys produce spiggin, a gluelike glycoprotein used in nest building, and its production is directly governed by androgens. Spiggin is normally absent in females but its production in female kidneys can be induced by AR agonists. Spiggin serves as the best known biomarker for androgens in fish. We exposed adult female sticklebacks to LNG at 5.5, 40, and 358 ng L(-1) for 21 days. Androgenic effects were found at LNG concentrations ≥40 ng L(-1) including induction of spiggin transcription, kidney hypertrophy, and suppressed liver vitellogenin transcription. These are the first in vivo quantitative data showing that LNG is a potent androgen in fish supporting the contention that androgenic effects of certain progestins contribute to their reproductive toxicity.

Short-term exposure to an endocrine disruptor affects behavioural consistency in male threespine stickleback

It has been established that exposure to xenooestrogens can have profound effects on reproductive and aggressive behaviours in aquatic organisms. Studies on the effects of 17α-ethinyloestradiol (EE2) on behaviour, however, rarely examine these effects over multiple trials, across time, or across contexts. In addition, it is possible that individuals vary in their sensitivity to EE2 and individuals may differ in the degree to which their behaviour is affected. The study of consistent individual differences in behaviour has grown in popularity in recent years perhaps because of the evolutionary questions this behavioural variation raises. Adult male threespine stickleback ( Gasterosteus aculeatus) from an anadromous population repeatedly received the simultaneous presentation of a dummy male and dummy female conspecific both before and after exposure to a short-term, nominal (15 ng L −1) concentration of 17α-ethinyloestradiol (EE2). These before and after measures were then compared against one another as well as against responses of males in a control group that was not exposed to EE2. Courtship, aggression, and nesting behaviours were all affected by EE2 exposure. In addition, behavioural consistency over trials was markedly reduced after EE2 exposure, as shown by lower repeatability values in the after exposure measures and in weaker correlations between the before and after exposure measures in the EE2 group compared to the control group. This suggests that male threespine stickleback vary in their sensitivity to EE2. This study is one of the first to examine the effects of short-term EE2 exposure on behaviour across trials and has important implications for population-level fitness consequences of xenooestrogens.

Repeatability of decision-making behaviour in male threespine stickleback Gasterosteus aculeatus: Effects of dummy vs. live stimuli

Abstract Individuals select from a number of behaviours when responding to various situations and the decisions they make may affect their fitness. The costs and benefits of these responses vary among individuals causing them to differ even in identical situations. One example of this type of situation is when territorial males encounter both a male and female simultaneously, generating a trade-off that likely leads to individual differences due to differing costs of various actions among males. This situation commonly occurs in threespine stickleback, Gasterosteus aculeatus. However, for selection to act effectively, individuals must behave in a consistent manner and measuring repeatability can aid in understanding how selection may shape such trade-offs. Males of this species exhibit consistent individual differences in their response to dummy males and females but it is unknown if patterns are similar when feedback from the stimuli is present. To assess this, male threespine stickleback were tested with dummy and live male and female conspecifics, presented separately and simultaneously. While the same trends were found regardless of stimulus type, males were more aggressive towards the live conspecifics than to the dummies. Repeatability values were similar within a treatment regardless of whether live or dummy conspecifics were used, suggesting that individuals show the same level of consistency. This study adds to our understanding of consistent individual differences by demonstrating that feedback may not affect responses to conflicting stimuli and that male threespine stickleback respond in a consistent manner to both dummy and live stimuli.

To eat or not to eat: egg-based assessment of paternity triggers fine-tuned decisions about filial cannibalism

Filial cannibalism occurs in many animal species ranging from insects to mammals, and is especially well described in teleost fishes. Numerous causes may lead to this behaviour, e.g. certainty of paternity. However, the cues males use to assess their paternity often remain unknown. One possible way to differentiate between own and foreign offspring is by using egg cues. Nevertheless, in egg-laying species, evidence for this is still scarce. In this study, male three-spined sticklebacks (Gasterosteus aculeatus), a fish with paternal care in which sneaking as well as filial cannibalism is common, were allowed to care for manipulated nests that contained different percentages of own fertilized eggs. After 7 days, embryo survival was determined. Furthermore, brood-caring as well as aggressive behaviour was measured daily. Clutches containing a higher proportion of foreign eggs were more likely to be completely cannibalized than clutches containing a lower proportion of foreign eggs, particularly when the clutch was laid early in the breeding season. However, the behavioural observations revealed no influence of paternity. The results show that paternity triggers filial cannibalism in sticklebacks and that males are able to evaluate their paternity using egg cues alone.

Dietary carotenoid availability and reproductive effort influence the age-related decline in performance

Elevated breeding effort is known to increase an individual’s rate of senescence, although the underlying mechanisms are not well understood. One possibility is that the ability to resist senescence is limited by the availability of antioxidants, which are necessary to mitigate the deleterious effects of oxidative stress thought to underlie the aging process. Susceptibility to oxidative stress is likely to be particularly high during reproduction, and so for a given level of reproductive effort, the rate of senescence should be fastest in those individuals with the poorest antioxidant capacity. We tested this hypothesis in an experimental study of breeding male three-spined sticklebacks (Gasterosteus aculeatus) held under a high or low reproduction regime and fed on either high or low levels of carotenoids (potentially limiting dietary compounds with antioxidant properties). Fish on the high reproduction regime and those on the low-carotenoid diet both showed an accelerated decline in sustained swimming performance (an indicator of locomotor senescence) compared with males on the low reproduction regime and high-carotenoid diet, respectively. The swimming performance of fish subjected to the high rate of reproduction, but fed a low-carotenoid diet, appeared to decline most rapidly, perhaps because of the additive effects of increased levels of reproduction-induced oxidative stress and lower antioxidant availability. These findings show that both dietary carotenoid intake and breeding effort can impact on the age-related decline in swimming performance and have important implications for female choice and the capacity of males with insufficient antioxidant defenses to adequately perform paternal care.

Dietary carotenoid availability, sexual signalling and functional fertility in sticklebacks

In species where males express carotenoid-based sexual signals, more intensely coloured males may be signalling their enhanced ability to combat oxidative stress. This may include mitigating deleterious oxidative damage to their sperm, and so be directly related to their functional fertility. Using a split-clutch in vitro fertilization technique and dietary carotenoid manipulation, we demonstrate that in non-competitive fertilization assays, male three-spined sticklebacks (Gasterosteus aculeatus) that are fed higher (but biologically relevant) levels of carotenoids had a significantly increased fertilization success, irrespective of maternal carotenoid intake. Furthermore, within diet groups, a male's fertilization success was positively related to the expression of his carotenoid-based nuptial coloration, with more intensely coloured males having higher functional fertility. These data provide, to our knowledge, the first demonstration that dietary access to carotenoids influences fertilization success, and suggest that females could use a male's nuptial coloration as an indicator of his functional fertility.

Further refinement of the non‐invasive procedure for measuring steroid production in the male three‐spined stickleback Gasterosteus aculeatus

Measurement of steroids that are released into the water via the gills has previously been shown to be an effective way of studying the reproductive endocrinology of the male three-spined stickleback Gasterosteus aculeatus without having to kill the fish. In the present paper, a previous observation on the existence of a compound other than 11-ketotestosterone (11-KT) in water, which cross-reacted in the 11-KT radioimmunoassay was repeated. The amounts of this compound, however, were not sufficient to warrant a separation step prior to carrying out assay. The lack of association between androstenedione levels in water and those in plasma was also confirmed. For the first time, the amounts of testosterone released into the water were shown to be positively correlated with the amounts in plasma, the sampling procedure (placing the fish for 30 min in 50 ml water) had no effect on the rate of release of cortisol but caused a rapid drop in the rate of release of 11-KT (which means that the fish should not be sampled twice in short succession), physical interaction between two nesting males (which was accompanied by aggression) significantly increased the rate of release of 11-KT, androstenedione and testosterone (but not of cortisol) and the rate of release of 11-KT was at its maximum between 2 and 4 h after exposure.

Reproductive potential of Schistocephalus solidus‐infected male three‐spined stickleback Gasterosteus aculeatus from two U.K. populations

Male three-spined stickleback Gasterosteus aculeatus from two U.K. populations with endemic infections of the cestode Schistocephalus solidus were brought into the laboratory prior to the breeding season and transferred to nesting tanks under conditions designed to stimulate sexual maturation. Nesting and courtship behaviours were scored over a 35 day period, after which fish were euthanized and the liver, spleen, kidney and gonads were weighed. Among G. aculeatus from a park pond in Leicester, U.K., infected males rarely engaged in reproductive behaviours and exhibited reduced indices of sexual development, body condition and general health, with effects being largely independent of relative parasite mass (parasite index, I(P)). In contrast, the reproductive behaviour of infected fish from Kendoon Loch in Dumfriesshire, U.K. appeared to be less severely affected, with infected fish regularly building nests and courting females under laboratory conditions. This was paralleled by a more limited effect of infection on physiological indicators of development, condition and general health. Furthermore, behavioural and physiological variables typically correlated with I(P) among infected fish from this population. Although comparing the performance of infected fish from the two populations directly was difficult due to potentially confounding factors, the results support the findings of recent studies showing that the effects of S. solidus on host reproduction are unlikely to be uniform across G. aculeatus populations. One possibility is that variation in the effects of infection arises from differences in the co-evolutionary association times of G. aculeatus with the parasite.

Nutritional benefits of filial cannibalism in three-spined sticklebacks (Gasterosteus aculeatus)

Although filial cannibalism (eating one's own offspring) occurs in numerous species, including several teleost fishes, its adaptive value is still not well understood. One often-discussed explanation is that individuals enhance their mass and body condition by consuming part of their eggs. However, evidence for this assumption is scarce thus far. In this study, male three-spined sticklebacks (Gasterosteus aculeatus), a species with paternal care, were allowed to care for a batch of eggs or for an empty nest under food-deprived conditions. All brood-caring males cannibalised at least part of their eggs and thus preserved their initial mass and body condition. Furthermore, mass as well as body condition was significant positively correlated with the number of cannibalised eggs. In contrast, empty-nest males that had no possibility to cannibalise eggs significantly lost mass and body condition. This is, to our knowledge, the first experimentally documented evidence that mass as well as body condition were preserved by filial cannibalism.

Detection of estrogenic activity in municipal wastewater effluent using primary cell cultures from three-spined stickleback and chemical analysis

Environmental estrogens are substances that imitate the effects of endogenous estrogens. Effluents from municipal wastewater treatment plants are known to contain substances with estrogenic activity including steroidal estrogens and xenoestrogens. In the current study, a combination of biological and chemical analysis was applied to determine the estrogenic activity in municipal wastewater effluents in Finland. The male three-spined stickleback ( Gasterosteus aculeatus) hepatocyte assay with vitellogenin induction as an endpoint was used for the detection of estrogenic activity in solid phase extracts of wastewater effluents, and 17β-estradiol (E2) as a positive control. The wastewater extracts and E2 were found to induce vitellogenin production. The extracts were also subjected to chromatographic fractionation and the collected fractions were assayed. The only active fraction was the one in which E2, estrone and ethynylestradiol were eluted. Its activity corresponded to the activity of the original wastewater extract. The LC–MS/MS analyses of the wastewater extracts showed that the concentration of estrone was about 65 ng L −1, the concentration of E2 was less than 1 ng L −1, while estriol and 17α-ethynylestradiol could not be detected. These findings showed that the activity of the wastewater extracts and the chromatographic fraction was much higher than the activity which could have been expected on the base of the chemical analysis. This strongly indicates that other compounds, possibly acting by additivity or synergism, are playing a major role in the induced vitellogenin production by the hepatocytes.